Pharmaceutical compositions and methods of treatment

Abstract

Provided are pharmaceutical compositions comprising a humanised anti-PSGL-1 IgG2 antibody of defined sequence, or antibody fragments or variants that share at least 90% identity with this antibody. Also relates to medical uses of the pharmaceutical compositions, and methods of treatment of diseases and conditions by administering such pharmaceutical compositions. The compositions incorporate the antibody at high concentration and may treat the diseases or conditions by decreasing inflammation.

Description

[0001] PHARMACEUTICAL COMPOSITIONS AND METHODS OF TREATMENT

[0002] FIELD OF INVENTION

[0003] The present invention relates to pharmaceutical compositions comprising a humanised anti- PSGL-1 lgG2 antibody, or antibody fragments or variants that share at least 90% identity with this antibody. The invention also relates to medical uses of the pharmaceutical compositions, and to methods of treatment of diseases and conditions, byadministering such pharmaceutical compositions. The compositions incorporate the antibody at high concentration and may treat the diseases or conditions by decreasing inflammation.

[0004] BACKGROUND

[0005] P-selectin glycoprotein ligand-1 (PSGL-1 ) is a component of the innate immune system and is found on leukocytes and endothelial cells. PSGL-1 binds to members of the selectin family of adhesion molecules including P-, E-, and L-selectin, but binds with highest affinity to P- selectin. PSGL-1 also binds key chemokines known to induce chemotaxis of cells into areas of inflammation.

[0006] PSGL-1 facilitates binding between leukocytes, platelets and the endothelium at sites of hypoxia, endothelial cell damage, and infection. Normally a beneficial process, when activation of these cells occurs unnecessarily or is excessive, PSGL-1 -mediated cell-cell interactions can cause a cascade of events culminating in inflammatory cell extravasation into tissues such as the lung parenchyma and bronchoalveolar space resulting in pathologic inflammation.

[0007] Anti-PSGL-1 antibodies are antibodies that bind to PSGL-1 . By binding to PSGL-1 anti-PSGL- 1 antibodies may inhibit the interaction of PSGL-1 with its ligands. This can prevent or reduce of cell adhesion (which may otherwise contribute to cell attachment, extravasation and migration) and may inhibit the PSGL-1 signalling pathway in cells. These activities render such antibodies promising therapeutic agents foruse in the prevention ortreatment of a range of diseases. Other reports have identified anti-PSGL-1 antibodies that exert an agonistic activity, down-regulating active T-cells without an impact on cell migration.

[0008] Asthma and chronic obstructive pulmonary disease (COPD) are common pulmonary inflammatory diseases characterized by excessive leukocyte infiltration into the lung parenchyma and bronchoalveolar space, resulting in tissue damage. Asthma is a common chronic inflammatory disease of the lungs characterized by episodes of wheezing, breathlessness, chest tightness, and cough, and the disease is often associated with eosinophil-rich airway inflammation. Airway eosinophilia is associated with asthma exacerbations and has been shown to play a role in airway remodelling. COPD represents one of the leading causes of death worldwide, and there are few effective therapies. Foreign particulate matter such as that produced from cigarette smoke leads to the recruitment of inflammatory cells to the bronchial wall and lumen resulting in progressive loss of lung function. Accumulation of neutrophils at sites of inflammation occurring in COPD may contribute to the pathophysiology of this disease, for example by secreting proteases that cause tissue destruction, or releasing mediators that further increase or prolong the inflammatory response.

[0009] VISTA (V-domain immunoglobulin suppressor of T-cell activation; also known as B7-H5, PD- 1H, Gi24, Dies1 , SISP1 and DD1α), is a B7 family ligand that is expressed on circulating and intratumoural myeloid cells and weakly expressed on activated lymphocytes. PSGL-1 binds to VISTA in low pH (acidic) environments, such as tumour beds.

[0010] It is generally recognised that infection, e.g. chronic infection (e.g., HIV, Hepatitis B, or Hepatitis C infection) and cancer can result in T-cell exhaustion. T-cell exhaustion is characterised by progressive loss of T-cell responsiveness, elimination of T-cells by apoptosis, and functional impairment and arrest of surviving T-cells. A loss of T-cell responsiveness in the context of infection, e.g. chronic infection, or cancer can thwart effective immune control. The mechanisms of T-cell exhaustion are incompletely understood, though it is often associated with an upregulation of immune co-inhibitory receptors (such as PD-1 , LAG3, CD160 and BTLA), loss of motility, altered transcriptional regulation, and expression of inhibitory cytokines (such as IL-10 and TGFβ). Reversal of T-cell exhaustion can therefore be linked to disease resolution against infection, e.g. chronic infection, and cancer, and this is an objective of cancer immunotherapies which utilise checkpoint inhibitor therapy.

[0011] Immune checkpoint inhibitors, i.e., drugs which target inhibitory receptors in the immune system (e.g., T-cells), have become an important tool in the treatment of cancer. In recent years, ipilimumab and nivolumab, antibodies targeting CTLA-4 and PD-1 respectively, have been developed and approved in the USA and Europe. However, many subjects with cancer do not respond to, or cease responding to, immune checkpoint inhibitor therapies. Therefore, there remains a need to identify new therapies which can complement or improve immune checkpoint inhibitor therapy, and / or reduce T-cell exhaustion. In the treatment of many diseases orconditions (including inflammatory diseases like asthma or COPD, and diseases such as cancer), subcutaneous administration presents numerous benefits. This route offers slower absorption compared to intravenous delivery, ensuring more consistent drug levels in the bloodstream. Additionally, in the context of inflammatorydiseases( it enables targeted delivery to subcutaneous tissue, which often correspondsto inflamed areas in these conditions.

[0012] Conventional high-concentration antibody formulations, especially those tailored for subcutaneous use, are notorious for encountering various critical challenges. These include viscosity issues, concerns regarding osmolarity, risks of aggregation, and potential reactions at the injection site.

[0013] Addressing these challenges is pivotal for the successful development and market introduction of high-concentration antibody formulations for subcutaneous administration. It necessitates innovative approaches in formulation design, manufacturing techniques, and the incorporation of stabilizing agents to guarantee the safety, efficacy, and patient acceptance of the final product.

[0014] SUMMARY OF THE INVENTION

[0015] According to a first aspect of the invention, there is provided a pharmaceutical composition comprising:

[0016] • an antibody at a concentration of approximately 210 mg / mL;

[0017] • 10mM acetate;

[0018] • 110mM sorbitol;

[0019] • 0.1 % poloxamer 188;

[0020] • 50mM proline and;

[0021] • 110mM mannitol;

[0022] • wherein the composition has a pH of 5.3; and

[0023] • the antibody comprises or consists of an antibody heavy chain of SEQ ID NO: 1 and an antibody light chain of SEQ ID NO: 2, or a fragment or variant of such an antibody sharing at least 90% sequence identity with an antibody that consists of SEQ ID NO: 1 and SEQ ID NO:2.

[0024] In a second aspect, the invention provides a pharmaceutical composition comprising:

[0025] • an antibody at a concentration of at least 150 mg / mL; • 9mM to 11 mM acetate;

[0026] • 50mM to 220mM sorbitol;

[0027] • 0.05% to 0.15% poloxamer 188;

[0028] • 45mM to 55mM proline; and

[0029] • 100mM to 220mM mM mannitol;

[0030] • wherein the composition has a pH of approximately 5.3; and

[0031] • the antibody comprises or consists of an antibody heavy chain of SEQ ID NO: 1 and an antibody light chain of SEQ ID NO: 2, or a fragment or variant of such an antibody sharing at least 90% sequence identity with an antibody that consists of SEQ ID NO: 1 and SEQ ID NO:2.

[0032] Suitably the antibody present in a pharmaceutical composition of the invention is the antibody designated “SelK2”, a humanised anti-PSGLantibody having the antibody heavy chain of SEQ ID NO: 1 and the antibody light chain of SEQ ID NO: 2, or a fragment or variant of SelK2 that shares at least 90% identity with SelK2.

[0033] Suitably the antibody present in a pharmaceutical composition of the invention comprises an antibody comprising SEQ ID NO: 1 and SEQ ID NO: 2. Suitably the antibody present in a pharmaceutical composition of the invention consists of an antibody comprising SEQ ID NO: 1 and SEQ ID NO: 2.

[0034] Suitably the antibody present in a pharmaceutical composition of the invention comprises an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2. Suitably the antibody present in a pharmaceutical composition of the invention consists of an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2.

[0035] In a suitable embodiment, the antibody present in a pharmaceutical composition in accordance with the invention comprisesan antibody, fragment, or variant sharing at least 95% sequence identity with an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2. Suitably, the antibody present in a pharmaceutical composition in accordance with the invention consists of an antibody, fragment, or variant sharing at least 95% sequence identity with an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2.

[0036] In a third aspect, the invention provides a method of preventing and / ortreating a disease or condition in a subject in need thereof, the method comprising providing to the subject a therapeutically effective amount of a pharmaceutical composition in accordance with the invention.

[0037] In a fourth aspect, the invention provides a pharmaceutical composition in accordance with the invention for use as a medicament.

[0038] In a suitable embodiment of the method in accordance with the third aspect of the invention, the composition is provided to the subject by subcutaneous injection. Similarly, in a suitable embodiment of the medical use of the fourth aspect of the invention, the composition is for use by subcutaneous injection.

[0039] In a suitable embodiment of the method in accordance with the third aspect of the invention, the disease or condition to be prevented and / or treated is an inflammatory disease or condition. Similarly, in a suitable embodiment of the medical use of the fourth aspect of the invention, the composition is for use to prevent and / or treat an inflammatory disease or condition.

[0040] In a fifth aspect, the invention provides a method for the preparation of a pharmaceutical composition comprising approximately 210 mg / mL SelK2 or a fragment or variant thereof, the method comprising combining:

[0041] • a solution comprising an antibody at a concentration of greater than 210 mg / ml in 10mM acetate, wherein the antibody comprises a heavy chain of SEQ ID NO: 1 and an antibody light chain of SEQ ID NO: 2, or a fragment or variant thereof sharing at least 90% sequence identity with SelK2; with

[0042] • 110mM sorbitol; 0.1 % poloxamer 188; 50mM proline; and 110 mM mannitol;

[0043] • to form a mixture comprising the antibody at a concentration of approximately 210 mg / mL; and

[0044] • sterilizing the mixture, to produce the pharmaceutical composition.

[0045] In a suitable embodiment, the method in accordance with the fifth aspect of the invention further comprises adjusting the pH to 5.3 using 1 % acetic acid. Suitably the sterilizing is by filtration, for example by using a 0.2 μm PES filter.

[0046] BRIEF DESCRIPTION OF THE FIGURES

[0047] FIG. 1 shows the results of a kinetic assay of SelSP1 binding to captured parental antibody. FIG. 2 shows the results of a kinetic assay of SelSP1 binding to captured SelK2 (high density surface).

[0048] FIG. 3 shows the results of a kinetic assay of SelSP1 binding to captured SelK2 (low density surface).

[0049] FIG. 4 shows the results of a study demonstrating the ability of SelK2 to inhibit neutrophil rolling.

[0050] FIG. 5 illustrates parental antibody and SelK2 antibody of the invention inhibiting chemokine CCL27 binding and interaction with PSGL-1. CCL27 complete assay cycle. Injection ‘A’ is surface blocking, injection ‘B’ is CCL27, injection ‘C’ is SDS, and injection ‘D’ is NaOH.

[0051] FIG. 6 illustrates the blocking of parental and SelK2 antibody zoomed into injection ‘B’ and re - normalized before injection. CCL27 (990 nM) injection. Solid line curves represent buffer blocked surface, dot-dash line curves represent K2 blocked surface and dashed line curves represent SelK2 blocked surface.

[0052] FIG. 7 illustrates SelK2 inhibition of human neutrophil binding to P-selectin under flow.

[0053] FIG. 8 shows the change in mean serum concentration of SelK2 over time (PK) after administration of two 7.5 mg / kg doses 21 days apart.

[0054] FIG. 9 illustrates the change in mean %Psel-lg Inhibition in response to SelK2 serum concentration over time (PD) after administration of two 7.5 mg / kg doses 21 days apart.

[0055] FIG. 10 shows the %Fall in FEV1 in placebo subjects during screening versus during treatment following allergen challenge (Mean ±SE).

[0056] FIG. 1 1 shows the %Fall in FEV1 at Day 36 (after 5 weeks of treatment) following allergen challenge in asthmatic subjects receiving placebo or SelK2 treatment in accordance with the invention (Mean ±SE).

[0057] FIG. 12 shows mean (±SE) absolute differential eosinophil count (10E6 / g) in sputum of asthmatic patients treated with placebo or SelK2 following allergen challenge. FIG. 13 shows mean (±SE) absolute eosinophils count (10E9 / L) in blood of asthmatic patients treated with placebo or SelK2 following allergen challenge.

[0058] FIG. 14 illustrates the mean (±SE) differential eosinophil count (%) in sputum of asthmatic patients treated with placebo or SelK2 following allergen challenge.

[0059] FIG. 15 illustrates the mean (±SE) differential eosinophil count (%) in blood of asthmatic patients treated with placebo or SelK2 following allergen challenge.

[0060] FIG. 16 shows mean (±SE) absolute differential eosinophil count (10E6 / g) in sputum of COPD patients treated with placebo or SelK2. Day 15, p=0.0494, Day 22, p=0.0181.

[0061] FIG. 17 shows mean (±SE) absolute differential eosinophil count (10E6 / L) in blood of COPD patients treated with placebo or SelK2.

[0062] FIG. 18 shows mean (±SE) absolute differential epithelial cell count (10E6 / g) in sputum of COPD patients treated with placebo or SelK2. P=0.0167

[0063] FIG. 19 compares %Fall in FEV1 following allergen challenge in patients with mild asthma treated with SelK2 versus Tezepelumab.

[0064] FIG. 20 shows mean (±SE) absolute numbers of eosinophils in sputum at baseline and day 22 of COPD patients treated with placebo or SelK2.

[0065] FIG. 21 shows mean (±SE) absolute numbers of neutrophils in sputum at baseline and day 22 of COPD patients treated with placebo or SelK2.

[0066] FIG. 22 shows mean (±SE) total number of cells per gram of sputum at baseline and day 22 of COPD patients treated with placebo or SelK2.

[0067] FIG. 23 shows results of Example 14, comparing expression of the T-cell effector molecule interferon-γ by peripheral blood mononuclear cells cultured in different experimental conditions.

[0068] DETAILED DESCRIPTION OF THE INVENTION The present invention is based upon the inventors’ development of a pharmaceutical composition that is able to incorporate therapeutically effective antibodies at high concentrations, while also having physical and chemical characteristics that ensure that the pharmaceutical composition is well tolerated when administered to a subject by injection. The skilled person will recognise that the development of high concentration antibody medicaments has historically proven difficult.

[0069] It will be appreciated that the ability to achieve high concentrations of therapeutic antibodies in the pharmaceutical compositions of the invention offers many notable advantages in practice. Compared to a lower concentration composition, a given volume of a pharmaceutical composition of the invention will provide a larger quantity of the active antibody. Alternatively, a comparable quantity of antibody may be administered in a smaller volume of the medicament.

[0070] As referred to above, antibody formulations of the present invention may incorporate a humanised anti-PSGL-1 antibody referred to herein as “SelK2”. The pharmaceutical compositions of the invention may also be used for the delivery of othertherapeuticantibodies that are fragments or variants sharing a high degree of sequence identity with SelK2. The similarity of these agents to the exemplary SelK2 antibody allows them to benefit from the same advantages in terms of stability and high concentration.

[0071] The pharmaceutical compositions of the invention are particularly well suited to administration by subcutaneous injection. Here the high concentrations of antibodies that may be incorporated in the pharmaceutical compositions of the invention are highly effective in providing a depot of the therapeutic antibody able to confer protection over a prolonged period of time.

[0072] Antibodies tend to aggregate and lose activity at high concentration, and many of the approaches that may be taken to prevent this yield products that are not suitable for clinical use.

[0073] In a significant breakthrough, the inventors have devised a high-concentration antibody composition (>200 mg / mL) that remarkably circumvents these conventional issues associated with high antibody formulations.

[0074] Formulating high-concentration antibody formulations, particularly for subcutaneous administration, presents considerable challenges well-recognized within the field. For present purposes, "high-concentration" of an antibody may be taken to refer to a formulation with antibody concentrations exceeding 150 mg / ml, for example at a concentration of approximately 150 to 250 mg / ml.

[0075] It will be appreciated that high-concentration formulations for injection, such as those of the present invention, are associated with a number of difficulties that are not relevant to high- concentration solutions, such as stock solutions, that are not intended to be administered.

[0076] One notable challenge is the heightened viscosity often associated with high-concentration antibody formulations. This elevated viscosity poses difficulties across various routes of administration, including subcutaneous and intravenous administration. High viscosity can impede the smooth injection of the formulation and may impact its distribution within the tissues or bloodstream post-administration.

[0077] Moreover, achieving and maintaining solubility of the antibody at such high concentrations presents a formidable hurdle. As antibody concentration rises, the risk of protein aggregation or precipitation escalates, potentially compromising the stability and efficacy of the formulation.

[0078] Physical instability further complicates matters for high-concentration antibody formulations. Issues such as protein aggregation, particle formation, or denaturation can arise during storage or upon injection. Ensuring stability throughout the product's shelf life and administration becomes paramount, posing significant technical challenges.

[0079] Antibodies are also susceptible to chemical degradation, including oxidation, deamidation, or hydrolysis, which are exacerbated at higher concentrations. Formulation optimization is essential to mitigate these degradation pathways and preserve the chemical stability of the antibody product.

[0080] Additionally, maintaining the biological activity of antibodies presents a critical consideration. Ensuring that antibodies retain their ability to bind to target antigens and exert therapeutic effects is paramount. However, achieving this balance is challenging, as formulation conditions such as pH, buffer composition, and excipients can influence antibody stability and activity.

[0081] These challenges contribute to the limited number of approved high-concentration antibody formulations, particularly in the United States, where, at the time the present invention has been made, only 17 formulations with antibody concentrations exceeding 150 mg / ml have received regulatory approval. Ofthese, merelyfive formulations boast antibody concentrations surpassing 200 mg / ml.

[0082] The inventors have identified a unique combination of excipients which allow for a high- concentration antibody formulation which comprises SelK2 at a final concentration of 210 mg / ml. The composition utilises excipients which are not commonly used in high concentration antibody formulations. As demonstrated in the Examples, the pharmaceutical compositions of the invention allow high concentrations of SelK2to be stably incorporated in a formulation that is well suited for administration to a subject.

[0083] As demonstrated in the Examples, SelK2, which has therapeutic utility in contexts such as the treatment of asthma or COPD. In at least some of these embodiments, the therapeutic function of the antibody arises from an ability to block cell adhesion, or interaction with ligands that contribute to cell attachment, extravasation and migration.

[0084] SelK2 and the closely related antibody fragments and variants considered herein may also function as antagonistic antibodies, able to inhibit the PSGL-1 signalling pathway in cells. This property, which is also demonstrated by the parental antibody from which SelK2 is derived, gives rise to a number of further therapeutic applications in respect of the pharmaceutical compositions of the invention.

[0085] For example, the disclosed antibodies may include an ability to reduce T-cell exhaustion. It will be readily appreciated that these propertiesmay confertherapeutic utility on the antibodies disclosed, and this gives rise to methods of treatment and medical uses employing the antibodies of the invention (or antigen-binding fragments thereof).

[0086] Furthermore, the properties of the antibodies of the invention clearly indicate that these benefits may be provided in respect of many other conditions characterised by PSGL-1 activation and signalling, such as T-cell exhaustion.

[0087] As noted above, it is generally recognised that infection, e.g. chronic infection (e.g., HIV Hepatitis B or Hepatitis C infection), orcancercan result in T-cell exhaustion. T-cell exhaustion is characterised by progressive loss of T-cell responsiveness, elimination of T-cells by apoptosis, and functional impairment and arrest of surviving T-cells. A loss of T-cell responsiveness in the context of infection , e.g. chronic infection, or cancer can thwart effective immune control. The mechanisms of T-cell exhaustion are incompletely understood, though it is often associated with an upregulation of immune co-inhibitory receptors (such as PD-1, LAG3, CD160 and BTLA), loss of motility, altered transcriptional regulation, and expression of inhibitory cytokines (such as IL-10 and TGFβ). Reversal of T-cell exhaustion can therefore be linked to disease resolution against infection, e.g. chronic infection, and cancer.

[0088] PSGL-1 is highly expressed on T-cells following chronic infection and knockout of PSGL-1 (Selplg- / -) in a chronic infection model has been found to result in increased T-cell accumulation, preservation of CD4+ and CD8+ T-cell effectors, production of pro-inflammatory cytokines, and diminished expression of several inhibitory receptors (Tinoco et al. 2016 Immunity 44, 1190-1203). Similarly, knockout of PSGL-1 prevents T-cell exhaustion and reduction of turn our growth in an aggressive melanoma model (Dankortetal. 2009 Nat. Genet. 41 , 544-552). Thus, PSGL-1 has been identified as a potent negative regulator of T-cell function in chronic infection and cancer.

[0089] VISTA (V-domain immunoglobulin suppressor of T-cell activation; also known as B7-H5, PD- 1H, Gi24, Diesl , SISP1 and DD1α) is a B7 family ligand that is expressed on circulating and intratumoural myeloid cells and weakly expressed on activated lymphocytes. The extracellular domain of VISTA is rich in histidine residues, of which the imidazole sidechain of histidine protonates at physiologically relevant pH. It has been found that VISTA preferentially engages and suppresses T-cells selectively in acidic environments such as tumour microenvironments, where pH values as low as 5.85 have been measured. It has also been found that VISTA binds PSGL-1 selectively at acidic pH via the extracellular histidine residues (Johnston et al. 2019 Nature 574; 565-570). Thus, VISTA is an acidic pH-selective ligand for PSGL-1 , i.e., VISTA may bind PSGL-1 in the tumour microenvironment.

[0090] Without wishing to be bound by any hypothesis, the ability of the antibodies suitable for use in the methods of treatment or medical uses of the invention to antagonise PSGL-1 signalling in cells may facilitate their ability to reduce or ameliorate T-cell exhaustion, thus stimulating the immune response. This impact indicates that these treatments have the potential to provide therapies for a broad range of diseases or conditions associated with T-cell exhaustion, such as cancer or infection, e.g. chronic infection.

[0091] Various aspects and embodiments of the present invention are now further described with reference to the following definitions.

[0092] Antibodies suitable for incorporation in pharmaceutical compositions of the invention The pharmaceutical compositions of the invention comprise a therapeutically effective antibody, fragment or variant. In particular, the pharmaceutical compositions of the invention may comprise a therapeutically effective antibody referred to herein as SelK2, or a therapeutically effective fragmentorvariant ofthisantibody. SelK2 is a humanised anti-PSGL- 1 antibody with a heavy chain having the amino acid sequence of SEQ ID NO: 1 and a light chain having the amino acid sequence of SEQ ID NO: 2. As described further elsewhere in this specification, SelK2 (and hence pharmaceutical compositions of the invention incorporating SelK2) has therapeutic utility in a wide range of applications.

[0093] Pharmaceutical compositions of the invention may incorporate any suitable therapeutically effective antibodies, or antibody fragments or variants, meeting the requirements set out herein. Such antibody fragments or variants may be antigen-binding antibody fragments or variants. For the purposes of the brevity in the present disclosure, except for where context requires otherwise, references to an “antibody” may also be taken as encompassing therapeutically effective fragments or variants of an antibody.

[0094] Suitable therapeutically effective antibodies, or their fragments or variants, that may be incorporated in the pharmaceutical compositions of the invention may be defined with reference to their degree of identity to an exemplary antibody. In particular, these fragments or variants may be defined with reference to their degree of identity to SelK2, the antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2. As described later, antibody fragments or variants sharing a high degree of identity with SelK2 may be able to provide similartherapeutic benefits to those that have been demonstrated in respect of SelK2. Accordingly, SelK2, and its fragments and variants may all be employed in the methods of treatment or medical uses of the invention contemplated herein.

[0095] For the present purposes, a “fragment” of an antibody should be taken as being a polypeptide that is truncated as compared to a reference antibody (such as SelK2), but which otherwise shares 100% identity to the amino acid sequence of the reference antibody. In contrast, a “variant” of an antibody should be taken as being a polypeptide the amino acid sequence of which contains at least one alteration as compared to the amino acid sequence of a reference antibody (such as SelK2). A variant of an antibody may have the same length as a reference antibody, or may be longer or shorter than the reference antibody.

[0096] Antibody fragments or variants suitable for incorporation in the pharmaceutical compositions of the invention share at least 90% identity with SelK2, the antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2. Percentage identity to a sequence, often referred to as “sequence identity”, is a measure used to quantify the degree of similarity between two biological sequences, such as DNA, RNA, or protein sequences. This measurement is expressed as a percentage and indicates how closely the sequences match each other at the same positions or residues. The skilled person will recognise that there are multiple common techniques that may be used to calculate percentage identity. Suitable techniques that may be used to calculate percentage identity include alignment of the sequences using various algorithms available for this purpose, such as BLAST (Basic Local Alignment Search Tool), ClustalW or specialized protein alignment software like MUSCLE or MAFFT followed by counting the matching positions, determining the total aligned positions and calculating the percentage identity using the following formula: Percentage Identity = (Number of Matching Positions / Total Aligned Positions) x 100.

[0097] Further details of suitable antibodies, fragments and variants that may be incorporated in the pharmaceutical compositions of the invention are provided elsewhere in the present disclosure.

[0098] SelK2 - an exemplary antibody suitable for incorporation in a pharmaceutical composition of the invention

[0099] SelK2 is a humanised anti-PSGL-1 lgG2 antibody that is an exemplar of the antibodies that may be incorporated in pharmaceutical compositions of the invention. As noted above, SelK2 consists of the antibody heavy chain set out in SEQ ID NO: 1 (the heavy chain variable region of which is set out in SEQ ID NO: 3) and the antibody light chain set out in SEQ ID NO: 2 (the light chain variable region of which is set out in SEQ ID NO: 4).

[0100] As illustrated in the Examples, SelK2 has a high affinity for PSGL-1 (KD of 4.28 nM, which is surprisingly strongerthan the parental antibody’s affinity of 16.4 nM). SelK2 is able to inhibit the formation of complexes of PSGL-1 and its ligands (including selectins, such as P-selectin L-selectin and E-selectin, chemokines, and VISTA) bound to each other, and is also able to disrupt such complexes of PSGL-1 and its ligands that have already formed. This activity underpins the therapeutic utility of SelK2 in certain embodiments of the invention directed at alleviating the symptoms of diseases or conditions, including inflammatory conditions (such as asthma and chronic obstructive pulmonary disease (COPD)), cancer, and infections (suitably chronic infections).

[0101] As well as pharmaceutical compositionscomprising SelK2, the present invention also provides pharmaceutical compositions comprising antibodies that are closely related to SelK2. Thus, the pharmaceutical compositions of the invention may comprise an antibody that comprises or consists of an antibody heavy chain of SEQ ID NO: 1 and an antibody light chain of SEQ ID NO: 2 (i.e. SelK2 or a modified form of SelK2), or a fragment orvariant of such an antibody sharing at least 90% sequence identity with an antibody that consists of SEQ ID NO: 1 and SEQ ID NO:2 (i.e. a therapeutically effective fragment of SelK2, or a therapeutically effective antibody that is very closely structurally related to SelK2).

[0102] An antibody, or an antigen-binding fragment thereof, in accordance with this second aspect of the invention may comprise the heavy chain amino acid sequence set out in SEQ ID NO: 1 and the light chain amino acid sequence set out in SEQ ID NO: 2. An antibody in accordance with this aspect of the invention may consist of the heavy chain amino acid sequence set out in SEQ ID NO: 1 and the light chain amino acid sequence set out in SEQ ID NO: 2.

[0103] As with SelK2, antibodies that may be incorporated in the pharmaceutical compositions of the invention may be humanised antibodies, or fragments or variants thereof.

[0104] As with SelK2, antibodies that may be incorporated in the pharmaceutical compositions of the invention may be antibodies, or fragments or variants thereof, that bind to PSGL-1 . Suitably, antibodies that may be incorporated in the pharmaceutical compositions of the invention may have the ability to bind to PSGL-1 with high affinity. Suitably such antibodies may have the ability to provide beneficial therapeutic activity through such binding.

[0105] Constituents of the pharmaceutical compositions of the invention

[0106] Formulating high concentration antibody compositions, particularly for subcutaneous administration, presents considerable challenges that are well-recognized within the field. For present purposes, "high concentration" of an antibody in a pharmaceutical composition may be taken to refer to a formulation with antibody concentrations exceeding 150 mg / ml, for example at a concentration of between approximately 150 and 250 mg / ml.

[0107] The inventors have identified a unique combination of excipients which enable the production of a high-concentration pharmaceutical composition comprising an antibody, such as SelK2. In a suitable embodiment the antibody, such as SelK2 is provided at a final concentration of 210 mg / ml.

[0108] As mentioned previously, the excipients utilised in the pharmaceutical compositions of the invention are not commonly used in high concentration antibody formulations, and the inventors believe that these contribute to the unexpectedly favourable properties possessed by the pharmaceutical compositions of the invention.

[0109] The first and second aspects of the invention provide compositions which comprise an antibody such as SelK2 (or or a fragment or variant thereof), in combination with acetate, sorbitol, poloxamer 188, proline and mannitol.

[0110] In a pharmaceutical composition in accordance with the first aspect of the invention, acetate is present at a concentration of 10 mM, sorbitol at a concentration of 110 mM, poloxamer 188 at approximately 0.1 %, proline at a concentration of 50 mM and mannitol at a concentration of 110 mM.

[0111] In a pharmaceutical composition in accordance with the second aspect of the invention, acetate is present at a concentration of 9 to 11 mM, sorbitol at a concentration of 50 to 220 mM, poloxamer 188 atapproximately0.05 to 0.15%, proline ata concentration of 45 to 55 mM and mannitol at a concentration of 100 to 220 mM. In a suitable embodiment of such a pharmaceutical composition of the second aspect of the invention, acetate is present at a concentration of approximately 10 mM, and / or sorbitol is present at a concentration of approximately 75 to 150 mM, and / or poloxamer is present at approximately 0.1 %, and / or proline is present at a concentration of approximately 50 mM, and / or mannitol is present at a concentration of approximately 100 mM to 150 mM.

[0112] Concentration of an antibody, fragment or derivative

[0113] Pharmaceutical compositions of the first aspect of the invention comprise an antibody, fragment or variant at a concentration of 210 mg / ml.

[0114] Pharmaceutical compositions of the second aspect of the invention comprise an antibody, fragment or variant at a concentration of at least 150 mg / ml. As noted above, this represents a high concentration of an antibody to be incorporated in a pharmaceutical composition. In a suitable embodiment, a pharmaceutical composition of the second aspect of the invention may comprise an antibody, fragment or variant at a concentration of up to 300 mg / ml. Other aspects of the invention provide pharmaceutical compositions that incorporate antibodies at a concentration of between 150 and 250 mg / ml.

[0115] In a suitable embodiment, a pharmaceutical composition of the second or sixth to eleventh aspects of the invention may comprise an antibody, fragment or variant at a concentration of up to 250 mg / ml. In a suitable embodiment, such a pharmaceutical composition of the invention, such as a pharmaceutical composition of the second aspect of the invention, may comprise an antibody, fragmentor variant at a concentration of between about 150 mg / ml and about 250 mg / ml. In a suitable embodiment, such a pharmaceutical composition of the invention may comprise an antibody, fragmentor variant at a concentration of between about 160 mg / ml and about 240 mg / ml. In a suitable embodiment, such a pharmaceutical composition of the invention may comprise an antibody, fragment or variant at a concentration of between about 170 mg / ml and about 230 mg / ml. In a suitable embodiment, such a pharmaceutical composition of the invention may comprise an antibody, fragment or variant at a concentration of between about 180 mg / ml and about 220 mg / ml. In a suitable embodiment, such a pharmaceutical composition of the invention may comprise an antibody, fragment or variant at a concentration of between about 195 mg / ml and about 225 mg / ml. In a suitable embodiment, such a pharmaceutical composition of the invention may comprise an antibody, fragment or variant at a concentration of between about 196 mg / ml and about 224 mg / ml. In a suitable embodiment, such a pharmaceutical composition of the invention may comprise an antibody, fragment or variant at a concentration of between about 197 mg / ml and about 223 mg / ml. In a suitable embodiment, such a pharmaceutical composition of the invention may comprise an antibody, fragmentor variant at a concentration of between about 198 mg / ml and about 222 mg / ml. In a suitable embodiment, such a pharmaceutical composition of the invention may comprise an antibody, fragmentor variant at a concentration of between about 199 mg / ml and about 221 mg / ml. In a suitable embodiment, such a pharmaceutical composition of the invention may comprise an antibody, fragment or variant at a concentration of between about 200 mg / ml and about 220 mg / ml. In a suitable embodiment, such a pharmaceutical composition of the invention may comprise an antibody, fragment or variant at a concentration of between about 201 mg / ml and about 219 mg / ml. In a suitable embodiment, such a pharmaceutical composition of the invention may comprise an antibody, fragment or variant at a concentration of between about 202 mg / ml and about 218 mg / ml. In a suitable embodiment, such a pharmaceutical composition of the invention may comprise an antibody, fragment or variant at a concentration of between about 203 mg / ml and about 217 mg / ml. In a suitable embodiment, such a pharmaceutical composition of the invention may comprise an antibody, fragment or variant at a concentration of between about 204 mg / ml and about 216 mg / ml. In a suitable embodiment, such a pharmaceutical composition of the invention may comprise an antibody, fragmentor variant at a concentration of between about 205 mg / ml and about 215 mg / ml. In a suitable embodiment, such a pharmaceutical composition of the invention may comprise an antibody, fragment or variant at a concentration of between about 206 mg / ml and about 214 mg / ml. In a suitable embodiment, such a pharmaceutical composition of the invention may comprise an antibody, fragment or variant at a concentration of between about 207 mg / ml and about 213 mg / ml. In a suitable embodiment, such a pharmaceutical composition of the invention may comprise an antibody, fragment or variant at a concentration of betweenabout 208 mg / ml and about 212 mg / ml. In a suitable embodiment, such a pharmaceutical composition of the invention may comprise an antibody, fragment or variant at a concentration of between about 209 mg / ml and about 211 mg / ml. In a suitable embodiment, such a pharmaceutical composition of the invention may ay comprise an antibody, fragment or variant at a concentration of about 210 mg / ml.

[0116] Acetate

[0117] A pharmaceutical composition of the first or second aspect of the invention com prises acetate. Acetate acts as a buffering agent that helps maintain the pH of an aqueous solution. In a suitable embodiment, acetate may be provided in the form of sodium acetate.

[0118] In a suitable embodiment, a pharmaceutical composition of the second aspect of the invention comprises between about 9mM and about 11 mM of acetate. In a suitable example, a pharmaceutical composition of the second aspect of the invention may comprise between about 9mm and about 11 mM, about 9.5mM and about 10.5mM, about 9m M and about 10mM or between about 10mM and about 11 mM of acetate.

[0119] In a suitable embodiment, a pharmaceutical composition of the second aspect of the invention comprises about 10mM of acetate. In a suitable example, a pharmaceutical composition of the second aspect of the invention may comprise about 9mM, about 9.5mM, about 10mM, about 10.5 mM or about 11 mM of acetate.

[0120] In a sixth aspect, the invention provides a pharmaceutical composition comprising:

[0121] • an antibody at a concentration of between approximately 150 and 250 mg / mL;

[0122] • a pH buffering agent;

[0123] • sorbitol at a concentration of 110 mM or approximately 110 mM;

[0124] • poloxamer 188 at 0.1 % or approximately 0.1 %;

[0125] • proline at a concentration of 50 mM or approximately 50 mM and;

[0126] • mannitol at a concentration of 110 mM or approximately 110 mM;

[0127] • wherein the composition has a pH of 5.3 or approximately 5.3; and

[0128] • the antibody comprises or consists of an antibody heavy chain of SEQ ID NO: 1 and an antibody light chain of SEQ ID NO: 2, or a fragment or variant of such an antibody sharing at least 90% sequence identity with an antibody that consists of SEQ ID NO: 1 and SEQ ID NO:2.

[0129] Sorbitol

[0130] A pharmaceutical composition of the first or second aspect of the invention com prises sorbitol. Sorbitol is a sugar alcohol stabilizer. Typically, a sugar alcohol stabilizer helps to preserve the stability of antibodies in a pharmaceutical composition by preventing their aggregation, denaturation, or degradation during storage or processing.

[0131] In a suitable embodiment, a pharmaceutical composition of the second aspect of the invention comprises between about 50mM and about 220mM of sorbitol. In a suitable example a pharmaceutical composition of the second aspect of the invention may comprise between about 60mm and about 210mM, about 70mM and about 200mM, about 80mM and about 190mM, about 90mM and about 180mM, about 100mM and about 170mM, about 110mM and about 160mM, about 120mM and about 150mM, or about 130mM and about 140mM or sorbitol.

[0132] In a suitable embodiment, a pharmaceutical composition of the second aspect of the invention comprises about 110mM or sorbitol. In a suitable example, a pharmaceutical composition of the second aspect of the invention may comprise about 100mM, about 101 mM, about 102mM, about 103mM, about 104mM, about 105mM, about 106mM, about 107mM, about 108mM, about 109mM, about 110mM, about 111 mM, about 112mM, about 113mM, about 114mM, about 115mM, about 116mM, about 117mM, about 118mM, about 119mM or about 120mM of sorbitol.

[0133] In a seventh aspect, the invention provides a pharmaceutical composition comprising:

[0134] • an antibody at a concentration of between approximately 150 and 250 mg / mL;

[0135] • acetate at a concentration of 10 mM or approximately 10 mM;

[0136] • a stabilizer;

[0137] • poloxamer 188 at 0.1 % or approximately 0.1 %;

[0138] • proline at a concentration of 50 mM or approximately 50 mM and;

[0139] • mannitol at a concentration of 110 mM or approximately 110 mM;

[0140] • wherein the composition has a pH of 5.3 or approximately 5.3; and

[0141] • the antibody comprises or consists of an antibody heavy chain of SEQ ID NO: 1 and an antibody light chain of SEQ ID NO: 2, or a fragment or variant of such an antibody sharing at least 90% sequence identity with an antibody that consists of SEQ ID NO: 1 and SEQ ID NO:2.

[0142] Suitably, a stabilizer employed in a pharmaceutical composition of the seventh aspect of the invention is a sugar alcohol stabilizer.

[0143] In an eighth aspect, the invention provides a pharmaceutical composition comprising:

[0144] • an antibody at a concentration of between approximately 150 and 250 mg / mL;

[0145] • acetate at a concentration of 10 mM or approximately 10 mM;

[0146] • a sugar alcohol stabilizer;

[0147] • poloxamer 188 at 0.1 % or approximately 0.1 %;

[0148] • proline at a concentration of 50 mM or approximately 50 mM and;

[0149] • wherein the composition has a pH of 5.3 or approximately 5.3; and

[0150] • the antibody comprises or consists of an antibody heavy chain of SEQ ID NO: 1 and an antibody light chain of SEQ ID NO: 2, or a fragment or variant of such an antibody sharing at least 90% sequence identity with an antibody that consists of SEQ ID NO: 1 and SEQ ID NO:2.

[0151] A composition in accordance with this aspect of the invention may optionally comprise first and second sugar alcohol stabilizers.

[0152] Poloxamer 188

[0153] A pharmaceutical composition of the first or second aspect of the invention comprises poloxamer 188. Poloxamer 188 is a surfactant. A surfactant is used in formulations for various purposes such as viscosity stabilisation. Poloxamer 188 is a non-ionic surfactant. A non-ionic surfactant is a type of surfactant that does not carry a charge on its hydrophilic head group and therefore has no net electrical charge in their formulations.

[0154] In a suitable embodiment, a pharmaceutical composition of the second aspect of the invention comprises between about 0.05% and about 0.15% of poloxamer 188. In a suitable example, a pharmaceutical composition of the second aspect of the invention may comprise between about 0.06% and about 1 .4%, about 0.07% and about 0.13%, about 0.08% and about 0.12% or about 0.09% and about 0.11 % of poloxamer 188. In a suitable embodiment, a pharmaceutical composition of the second aspect of the invention comprisesaboutO.1 %of poloxamerl 88. In a suitable example, a pharmaceutical composition of the second aspect of the invention may comprise about 0.05%, about0.06%, about 0.07%, about 0.08%, about 0.09%, about 0.1 %, about 0.11 %, about 0.12%, about 0.13%, about 0.14% or about 0.15% of poloxamer 188.

[0155] In a ninth aspect, the invention provides a pharmaceutical composition comprising:

[0156] • an antibody at a concentration of between approximately 150 and 250 mg / mL;

[0157] • acetate at a concentration of 10 mM or approximately 10 mM;

[0158] • sorbitol at a concentration of 110 mM or approximately 110 mM;

[0159] • a surfactant;

[0160] • proline at a concentration of 50 mM or approximately 50 mM and;

[0161] • mannitol at a concentration of 110 mM or approximately 110 mM;

[0162] • wherein the composition has a pH of 5.3 or approximately 5.3; and

[0163] • the antibody comprises or consists of an antibody heavy chain of SEQ ID NO: 1 and an antibody light chain of SEQ ID NO: 2, or a fragment or variant of such an antibody sharing at least 90% sequence identity with an antibody that consists of SEQ ID NO: 1 and SEQ ID NO:2.

[0164] Suitably the surfactant employed in a pharmaceutical composition of the ninth aspect of the invention is a non-ionic surfactant.

[0165] Proline

[0166] A pharmaceutical composition of thefirstorsecond aspect of the invention comprises proline. Proline is a nonpolar, aliphatic amino acid. Typically, proline can help stabilize antibodies during various stages of formulation, storage, and handling.

[0167] In a suitable embodiment, a pharmaceutical composition of the second aspect of the invention comprises between about 45mM and about 55mM of proline. In a suitable example, a pharmaceutical composition of the invention may comprise between about 46mM and about 54mM, about 47mM and about 53mM, about 48mM and about 52mM, or between about 49mM and about 52mM or proline.

[0168] In a suitable embodiment, a pharmaceutical composition of the second aspect of the invention comprises about 50mM of proline. In a suitable example, a pharmaceutical composition of the invention may comprise about 45mM, about 46mM, about 47mM, about 48mM, about 49mM, about 50mM, about 51 mM, about 52mM, about 53mM, about 54mM, or about 55mM of proline.

[0169] In a tenth aspect, the invention provides a pharmaceutical composition comprising:

[0170] • an antibody at a concentration of between approximately 150 and 250 mg / mL;

[0171] • acetate at a concentration of 10 mM or approximately 10 mM;

[0172] • sorbitol at a concentration of 110 mM or approximately 110 mM;

[0173] • poloxamer 188 at 0.1 % or approximately 0.1 %;

[0174] • an amino acid stabilizer; and;

[0175] • mannitol at a concentration of 110 mM or approximately 110 mM;

[0176] • wherein the composition has a pH of 5.3 or approximately 5.3; and

[0177] • the antibody comprises or consists of an antibody heavy chain of SEQ ID NO: 1 and an antibody light chain of SEQ ID NO: 2, or a fragment or variant of such an antibody sharing at least 90% sequence identity with an antibody that consists of SEQ ID NO: 1 and SEQ ID NO:2.

[0178] Mannitol

[0179] A pharmaceutical composition of the first or second aspect of the invention comprises mannitol. Mannitol is a sugar alcohol stabilizer. Typically, a sugar alcohol stabilizer help preserve the stability of antibodies by preventing aggregation, denaturation, or degradation during storage or processing.

[0180] In a suitable embodiment, a pharmaceutical composition of the second aspect of the invention comprises between about 100mM and about 220mM of mannitol. In a suitable example, a pharmaceutical composition of the invention may comprise between about 110mM and about 210mM, about 120mM and about 200mM, about 130mM and about 190mM, about 140mM and about 180mM or about 150mM and about 170mM of mannitol.

[0181] In a suitable embodiment, a pharmaceutical composition of the second aspect of the invention comprises about 110mM of mannitol. In a suitable example, a pharmaceutical composition of the invention may comprise about 100mM, about 101 mM, about 102mM, about 103mM, about 104mM, about 105mM, about 106mM, about 107mM, about 108mM, about 109mM, about 110mM, about 111 mM, about 112mM, about 113mM, about 114mM, about 115mM, about 116mM, about 117mM, about 118mM, about 119mM or about 120mM of mannitol. In an eleventh aspect, the invention provides a pharmaceutical composition comprising:

[0182] • an antibody at a concentration of between approximately 150 and 250 mg / mL;

[0183] • acetate at a concentration of 10 mM or approximately 10 mM;

[0184] • sorbitol at a concentration of 110 mM or approximately 110 mM;

[0185] • poloxamer 188 at 0.1 % or approximately 0.1 %;

[0186] • proline at a concentration of 50 mM or approximately 50 mM and;

[0187] • a stabilizer;

[0188] • wherein the composition has a pH of 5.3 or approximately 5.3; and

[0189] • the antibody comprises or consists of an antibody heavy chain of SEQ ID NO: 1 and an antibody light chain of SEQ ID NO: 2, or a fragment or variant of such an antibody sharing at least 90% sequence identity with an antibody that consists of SEQ ID NO: 1 and SEQ ID NO:2.

[0190] The stabilizer employed in a pharmaceutical composition of the eleventh aspect of the invention may suitably be a sugar alcohol stabilizer.

[0191] Characteristics of pharmaceutical compositions of the invention pH

[0192] The stability of antibodies is highly dependent on pH. Antibodies have specific pH ranges at which they remain stable and maintain their native conformation and activity.

[0193] A pharmaceutical composition of the first aspect of the invention has a pH of 5.3, and a pharmaceutical composition of the second or sixth to eleventh aspects of the invention has a pH of 5.3 or approximately 5.3.

[0194] In a suitable example, a pharmaceutical compositionof the second orsixth to eleventh aspects of the invention may have a pH of about 5.2, about 5.3 or about 5.4.

[0195] Osmolality

[0196] Osmolality refers to the concentration of osmotically active particles in a solution per unit mass of solvent, typically measured in milliosmoles per kilogram (mOsmo / kg). It considers the total number of solute particles relative to the mass of the solvent, regardless of the solution's volume. Osmolality is not affected by changes in volume, making it an accurate measure in situations where volume changes occur, such as in biological systems.

[0197] The osmolality of a pharmaceutical composition, such as a composition comprising an antibody, is important for various reasons. For example, osmolality influences the stability of antibodies in solution. Deviations from physiological osmolality can lead to protein denaturation, aggregation, or precipitation, compromising the efficacy and safety of the antibody formulation. This serves to also preserve the functional integrity of antibodies.

[0198] Osmolality of a pharmaceutical composition becomes increasingly important when the composition is formulated for subcutaneous administration. For example, compositions with high or low osmolality can cause irritation or damage to the subcutaneous tissue. Solutions with osmolality within physiological ranges are less likely to cause discomfort or pain upon injection compared to solutions with more extreme osmolalities. Additionally, osmolality can affect the rate and extent of absorption of compositions administered.

[0199] In a suitable embodiment, a pharmaceutical composition of the invention has an osmolality between about 270 mOsmo / kg and about 310 mOsmo / kg. In a suitable example, a pharmaceutical composition of the invention may have an osmolality of between about 275 mOsmo / kg and about 305 mOsmo / kg, about 280 mOsmo / kgand about 300 mOsmo / kg, about 280 mOsmo / kg and about 295 mOsmo / kg orabout 285 mOsmo / kg and about 290 mOsmo / kg. In a suitable embodiment, a pharmaceutical composition of the invention has an osmolality of approximately 290 mOsmo / kg.

[0200] In a suitable embodiment, a pharmaceutical composition of the invention has an osmolality of 310 mOsmo / kg or less. In a suitable example, a pharmaceutical composition of the invention may have an osmolality of 300 mOsmo / kg or less, 290 mOsmo / kg or less, 280 mOsmo / kg or less or 270 mOsmo / kg or less.

[0201] In a suitable embodiment, a pharmaceutical composition of the invention has an osmolality of at least 270 mOsmo / kg. In a suitable example, a pharmaceutical composition of the invention may have an osmolality of at least 280 mOsmo / kg, at least 290 mOsmo / kg, at least 300 mOsmo / kg or at least 310 mOsmo / kg.

[0202] In a suitable embodiment, a pharmaceutical composition of the invention has an osmolality of about 310 mOsmo / kg. In a suitable example, a pharmaceutical composition of the invention may have an osmolality of about 300 mOsmo / kg, about 301 mOsmo / kg, about 302 mOsmo / kg, about 303 mOsmo / kg, about 304 mOsmo / kg, about 305 mOsmo / kg, about 306 mOsmo / kg, about 307 mOsmo / kg, about 308 mOsmo / kg, about 309 mOsmo / kg, about 310 mOsmo / kg, about 311 mOsmo / kg, about 312 mOsmo / kg, about 313 mOsmo / kg, about 314 mOsmo / kg, about 315 mOsmo / kg, about 316 mOsmo / kg, about 317 mOsmo / kg, about 318 mOsmo / kg, about 319 mOsmo / kg or about 320 mOsmo / kg.

[0203] It will be appreciated that osmolality of a pharmaceutical composition of the invention may be measured using any suitable technique. A person skilled in the art would be aware of many such suitable techniques, such as freezing point depression, vapor pressure osmometry, osmometry with osmometer or calculated osmolality.

[0204] Absolute viscosity

[0205] Absolute viscosity refers to the measure of the resistance of a composition to flow under applied stress, typically expressed in units of centipoise (cP) or millipascal-seconds (mPas).

[0206] In a suitable embodiment, a pharmaceutical composition of the invention has an absolute viscosity of 20 cP or less. In a suitable example, a pharmaceutical composition of the invention may have an absolute viscosity of 19 cP or less, 18 cP or less, 17 cP or less, 16 cP or less, 15 cP or less, 14 cP or less, 13 cP or less, 12 cP or less, 11 cP or less, 10 cP or less, 9 cP or less, 8 cP or less, 7 cP or less, 6 cP or less or 5 cP or less.

[0207] In a suitable embodiment, a pharmaceutical composition of the invention has an absolute viscosity of about 15 cP. In a suitable embodiment, a pharmaceutical composition of the invention may have an absolute viscosity of about 10 cP, about 11 cP, about 12 cP, about 13 cP, about 14 cP, about 15 cP, about 16 cP, about 17 cP, about 18 cP, about 19 cP or about 20 cP.

[0208] It will be appreciated that absolute viscosity of a pharmaceutical composition of the invention may be measured using any suitable technique. A person skilled in the art would be aware of many such suitable techniques, such as use of a viscometer or rheometer, dynamic light scattering (DLS), concentration dependence or shear rate dependence.

[0209] Subvisible particles Subvisible particles refers to particles within a formulation that are too small to be easily observed by the naked eye but are large enough to potentially affect product quality, safety, or efficacy. These particles typically range in size from approximately 2 to 100 micrometers (μm).

[0210] The number of subvisible particles present is an important consideration for a pharmaceutical composition for various reasons. Subvisible particles, especially protein aggregates or other foreign materials, can trigger immune responses in patients, leading to adverse reactions or reduced therapeutic effectiveness.

[0211] The USP 787 standard requires pharmaceutical and biopharmaceutical manufacturers making injections and infusionsfollow rigorous rules regarding the quantity of particles present in final drug products. The standard requires a formulation to meet the subvisible particle requirements of ≤ 6000 counts for ≥ 10 μm particles and ≤ 600 counts for ≥ 25 μm particles.

[0212] In a suitable embodiment, a pharmaceutical composition of the invention has less than 6000 counts for subvisible particles greater than 10 μm. In a suitable example, a pharmaceutical composition of the invention may have less than 5500 counts, less than 5000 counts, less than 4500 counts, less than 4000 counts, less than 3500 counts, less than 3000 counts, less than 2500 counts, less than 2000 counts, less than 1500 counts, less than 1000 counts or less than 500 counts for subvisible particles greater than 10 μm.

[0213] Alternatively, or in addition, in a suitable embodiment, a pharmaceutical composition of the invention has less than 600 counts for particles greater than 25 μm. In a suitable example, a pharmaceutical composition of the invention may have less than 550 counts, less than 500 counts, lessthan 450 counts, lessthan 400 counts, lessthan 350 counts, lessthan 300 counts, less than 250 counts, less than 200 counts, less than 150 counts or less than 100 counts for particles greater than 25 μm.

[0214] In a suitable embodiment, a pharmaceutical composition of the invention has about 500 counts for subvisible particles greater than 10 μm. In a suitable example, a pharmaceutical composition of the invention may have about 0 counts, about 100 counts, about 200 counts, about 300 counts, about 400 counts, about 500 counts, about 600 counts, about 700 counts, about 800 counts, about 900 counts or about 1000 counts forsubvisible particles greater than 10 μm.

[0215] Alternatively, or additionally, a pharmaceutical composition of the invention has about 300 counts for subvisible particles greater than 25 μm. In a suitable example, a pharmaceutical composition of the invention may have about 0 counts, about 100 counts, about 200 counts, about 300 counts, about 400 counts, about 500 counts or about 600 counts for subvisible particles greater than 25 μm.

[0216] In a suitable embodiment, a pharmaceutical composition of the invention has less than 6000 counts for subvisible particles greaterthan 10 μm and / or less than 600 counts for particles up to 25 μm.

[0217] It will be appreciated that any suitable technique may be used to measure the number of subvisible particles in a pharmaceutical composition. A person skilled in the art would be aware of many such suitable techniques, such as microscopy, flow imaging analysis, dynamic light scattering (DLS) or microscopic particle counting.

[0218] Injection glide force

[0219] In the context of injectable pharmaceutical compositions, injection glide force refers to the force required to move the plunger or piston within a syringe barrel during the injection process. Injection glide force is typically measured in units of force, such as Newtons (N) or pounds-force (Ibf).

[0220] Injection glide force is particularly important to consider when a pharmaceutical composition is to be administered subcutaneously, which is a relevant route of administration of the pharmaceutical compositions of the invention. Excessive injection glide force can cause discomfort or pain to the patient during administration. Furthermore, high injection glide force may lead to tissue damage or injection site reactions, such as bruising, swelling, or pain. Reducing the injection glide force can enhance patient comfort and compliance with treatment regimens.

[0221] In a suitable embodiment, a pharmaceutical composition of the invention has an injection glide force of 25 N or less. In a suitable example, a pharmaceutical composition of the invention may have an injection glide force of 20 N or less, 19 N or less, 18 N or less, 17 N or less, 16 N or less, 15 N or less, 14 N or less, 13 N or less, 12 N or less, 11 N or less, 10 N or less, 9 N or less, 8 N or less, 7 N or less, 6 N or less or even 5 N or less.

[0222] In a suitable embodiment, a pharmaceutical composition of the invention has an injection glide force of about 20 N. In a suitable example, a pharmaceutical composition of the invention may have an injection glide force of about 15 N, about 16 N, about 17 N, about 18 N, about 19 N, about 20 N, about 21 N, about 22 N, about 23 N, about 24 N or about 25 N.

[0223] The values in the preceding paragraphs may, for example, be determined in respect of injection of the pharmaceutical composition via a 25 G needle. The skilled person will be able to determine suitable values, based on those set out above and in the Examples, for other gauges of needle.

[0224] It will be appreciated that any suitable technique may be used to measure the injection glide force of a pharmaceutical composition. A person skilled in the art would be aware of many such suitable techniques, such as force sensors, electronic syringe drivers or digital force gauges.

[0225] Functional properties of suitable antibodies, fragments and variants

[0226] The antibody SelK2 (consisting of SEQ ID NO: 1 and SEQ ID NO:2) has functional properties that give rise to a number of therapeutic applications in which this antibody may be used. It will be appreciated that pharmaceutical compositions of the invention comprising SelK2 will also exhibit these advantageous functional properties, and so will be capable of conferring the therapeutic benefits of SelK2. Indeed, the high concentrations of SelK2 that can be delivered via the pharmaceutical compositions of the invention promise to provide excellent therapeutic utility in practice.

[0227] Other antibodies fragments and variants that may be incorporated in pharmaceutical compositions of the invention share a high degree of identity with SelK2. Given the close relationship between antibody structure and function (and the ways in which it is known that parts of antibody structure may be modified to either retain or change biological properties of a reference antibody) it will be apparent that many of these antibody fragments or variants may be expected to share similar biological, and hence therapeutic, properties with SelK2.

[0228] Suitably, antibodies that may be incorporated in a pharmaceutical composition of the invention may share SelK2’s ability to function as blocking antibodies, able to inhibit formation of complexes between PSGL-1 and its ligands, such as P-selectin. Antibodies suitable for incorporation in the pharmaceutical compositions of the invention may alternatively or additionally be able to disrupt complexes between PSGL-1 and its ligands, such as P-selectin, that have already formed. As discussed further below, such ability to inhibit or disrupt complexes of PSGL-1 and its ligands, such as P-selectin, may confer therapeutic utility on the antibodies to be incorporated in such compositions, and thus on the pharmaceutical compositions themselves, as well as methods of treatment and medical uses employing said pharmaceutical compositions.

[0229] Properties of SelK2 that may be conferred upon pharmaceutical compositions of the invention comprising this antibody, or which may be retained by suitable fragments or variants of antibodies to be incorporated in pharmaceutical compositions of the invention are discussed further below.

[0230] Affinity

[0231] SelK2 binds to PSGL-1 with high affinity. Since antibody fragments or variants suitable for incorporation in the pharmaceutical compositions of the invention are closely structurally related to SelK2, they may share this high affinity.

[0232] Humanised antibodies often exhibit lessened affinity for their target as compared to the parental antibody from which they are derived, particularly when germline sequences are utilised as framework acceptors. In contrast, the inventors have demonstrated that the SelK2 has a higher affinity for PSGL-1 than the parental antibody from which this humanised antibody was derived. The parental antibody from which SelK2 is derived has an affinity constant for PSGL-1 of 16.6 nM, whereas SelK2 has an affinity constant for PSGL-1 of 4.29 nM. This represents an almost four-fold increase in affinity as compared to the parental antibody, highly unusual in respect of a humanized antibody employing germline framework acceptor sequences.

[0233] In a suitable embodiment, an antibody fragment or variant incorporated in a pharmaceutical composition of the invention may similarly have an affinity constant for PSGL-1 of below 16 nM. For example, such an antibody fragment or variant may have an affinity constant for PSGL-1 of below 15 nM, of below 14 nM, of below 13 nM, of below 12 nM, of below 11 nM, of below 10 nM, of below 9 nM, of below 8 nM, of below 7 nM, of below 6 nM, or an affinity constant of below 5 nM. Suitably, such an antibody fragment or variant may have an affinity constant for PSGL-1 of approximately 4 nM, for example an affinity constant for PSGL-1 of 4.29 nM. In a suitable embodiment, an antibody fragment or variant incorporated in a pharmaceutical composition of the invention may have even greater affinity for PSGL-1 , for example an affinity constant of 4 nM or less, of 2 nM or less, or of 1 nM or less. The high affinity of SelK2 means that this antibody is able to not only inhibit binding of PSGL- 1 to its ligands, but also to disrupt such binding that has already occurred. Antibodies, fragments or variants sharing a high degree of identity with SelK2 may also share a high affinity for PSGL-1 , as set out in the preceding paragraph. As described further below, this opens new therapeutic windows for compositions of the invention which comprise Selk2, or closely related antibodies, fragments orvariants in which treatments can achieve benefit even when a response driven by binding of PSGL-1 to its ligand has already begun.

[0234] Accordingly, the pharmaceutical compositions, methods of treatment, or medical uses of the present invention comprising SelK2, or closely related antibody fragments or variants, will be able to not only inhibit binding of PSGL-1 to its ligands, but also to disrupt such binding that has already occurred.

[0235] The affinity for PSGL-1 of an antibody to be incorporated in a pharmaceutical composition of the invention may be determined by any suitable method known to one skilled in the art. Merely by way of example, affinity, such as the affinity values set out in the paragraph above, may be derived by Surface Plasmon Resonance (SPR) using a series of escalating concentrations through saturation. Details of suitable techniques, and the results achieved using these in respect of the exemplary antibody SelK2 are set out in the Examples.

[0236] Inhibition or disruption of binding of PSGL-1 to its ligands

[0237] As demonstrated in the Examples, SelK2 is able to inhibit and / or to disrupt binding of PSGL- 1 to its ligands. Suitably, an antibody fragment or variant to be incorporated in a pharmaceutical composition of the invention may share this ability to inhibit and / or disrupt binding of PSGL-1 to its ligands.

[0238] In the context of the present invention, “inhibition” of binding of PSGL-1 to a ligand may be taken as referring to the prevention of the formation of new interactions between PSGL-1 and the ligand in question.

[0239] On the other hand, “disruption” of binding of PSGL-1 to a ligand may be taken as referring to the breaking down of previously formed interactions between PSGL-1 and the ligand in question.

[0240] As discussed further elsewhere in this disclosure, these capabilities of SelK2 mean that compositions of the invention comprising SelK2, or closely related antibody fragments or variants, are well suited to both prevention of diseases or conditions caused by deleterious binding of PSGL-1 to its ligands, and to the treatment of diseases or conditions caused by deleterious binding of PSGL-1 to its ligands.

[0241] Generally, an antibody, fragment or variant suitable for incorporation in a pharmaceutical composition of the invention may be able to inhibit or disrupt binding of PSGL-1 to a ligand by at least 10%, at least 11 %, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least 21 %, at least 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, at least 28%, at least 29%, at least 30%, at least 31 %, at least 32%, at least 33%, at least 34%, at least 35%, at least 36%, at least 37%, at least 38%, at least 39%, at least 40%, at least 41 %, at least 42%, at least 43%, at least 44%, at least 45%, at least 46%, at least 47%, at least 48%, at least 49%, at least 50%, at least 51 %, at least 52%, at least 53%, at least 54%, at least 55%, at least 56%, at least 57%, at least 58%, at least 59%, at least 60%, at least 61 %, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71 %, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or even by 100%.

[0242] The skilled person will readily be able to identify suitable assays for determining an extent of inhibition or disruption of binding achieved. The skilled person will recognise that there are multiple common techniques that may be used to determine the extent of inhibition or disruption of binding achieved. Suitable techniques that may be used to determine the extent of inhibition or disruption of binding achieved include Enzyme-Linked Immunosorbent Assay (ELISA), Surface Plasmon Resonance (SPR), flow cytometry, western blotting, immunoprecipitation (IP) and competition binding assays. These may be selected with reference to particular ligands of interest (forexample, binding of PSGL-1 to P-selectin or to a chemokine), or with reference to particular binding-associated activities of interest (for example, cell rolling or extravasation).

[0243] A suitable pharmaceutical composition of the invention may have the ability to inhibitor disrupt binding of PSGL-1 to its ligands in accordance with any of the parameters set out above.

[0244] Accordingly, the pharmaceutical compositions of the invention may be used to inhibit or disrupt binding of PSGL-1 to ligands. Inhibition or disruption of binding to selectins

[0245] PSGL-1 is known to bind to selectins including those selected from the group consisting of: P- selectin, E-selectin and L-selectin.

[0246] As set out in the Examples, the inventors have demonstrated that SelK2 is able to inhibit or disrupt binding of PSGL-1 to specific selectin ligands, in particular P-selectin and L-selectin. Suitably, an antibody fragment or variant to be incorporated in a pharmaceutical composition of the invention may share this ability to inhibit or disrupt binding of PSGL-1 to such selectin ligands. Inhibition or disruption of such binding will prevent or reduce selectin-mediated biological activities.

[0247] Antibodies, or fragments or variants of antibodies, suitable for incorporation in the pharmaceutical compositions of the invention may be able to inhibit or disrupt binding of PSGL-1 to a selectin (such as P-selectin or L-selectin) by at least 10%, at least 11 %, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least 21 %, at least 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, at least 28%, at least 29%, at least 30%, at least 31 %, at least 32%, at least 33%, at least 34%, at least 35%, at least 36%, at least 37%, at least 38%, at least 39%, at least 40%, at least 41 %, at least 42%, at least 43%, at least 44%, at least 45%, at least 46%, at least 47%, at least 48%, at least 49%, at least 50%, at least 51 %, at least 52%, at least 53%, at least 54%, at least 55%, at least 56%, at least 57%, at least 58%, at least 59%, at least 60%, at least 61 %, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71 %, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or even by 100%.

[0248] The ability of an antibody, or fragment or variant thereof, incorporated in a pharmaceutical composition of the invention to inhibit or disrupt binding of PSGL-1 to a selectin ligand (such as P-selectin or L-selectin) may be investigated by any appropriate means. For instance, this may be assessed by cell-based assays, of the sort discussed in Examples 2 or 4. A pharmaceutical composition of the invention may have the ability to inhibit or disrupt binding of PSGL-1 to selectins in accordance with any of the parameters set out above.

[0249] Accordingly, the pharmaceutical compositions of the invention may be used to inhibit or disrupt binding of PSGL-1 to selectins.

[0250] Inhibition or disruption of binding to chemokines

[0251] PSGL-1 is known to bind to chemokines including those selected from the group consisting of: CCL27; CCL19; and CCL21.

[0252] As set out in the Examples, the inventors have demonstrated that SelK2 is able to inhibit or disrupt binding of PSGL-1 to chemokine ligands, as exemplified by CCL27. Suitably, an antibody fragment or variant to be incorporated in a pharmaceutical composition of the invention may share this ability to inhibit or disrupt binding of PSLG-1 to such chemokine ligands. Inhibition or disruption of such binding will prevent or reduce chemokine-mediated biological activities.

[0253] Antibodies, or fragments or variants of antibodies, suitable for incorporation in the pharmaceutical compositions of the invention may be able to inhibit or disrupt binding of PSGL-1 to a chemokine (such as CCL27) by at least 10%, at least 11 %, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least 21 %, at least 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, at least 28%, at least 29%, at least 30%, at least 31 %, at least 32%, at least 33%, at least 34%, at least 35%, at least 36%, at least 37%, at least 38%, at least 39%, at least 40%, at least 41 %, at least 42%, at least 43%, at least 44%, at least 45%, at least 46%, at least 47%, at least 48%, at least 49%, at least 50%, at least 51 %, at least 52%, at least 53%, at least 54%, at least 55%, at least 56%, at least 57%, at least 58%, at least 59%, at least 60%, at least 61 %, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71 %, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or even by 100%. The ability of an antibody, or fragment or variant thereof, incorporated in a pharmaceutical composition of the invention to inhibit or disrupt binding of PSGL-1 to a chemokine ligand (such as CCL27) may be investigated by any appropriate means. For instance, this may be assessed by a SPR-based assay, of the sort discussed in Example 3.

[0254] A pharmaceutical composition of the invention may have the ability to inhibit or disrupt binding of PSGL-1 to chemokines in accordance with any of the parameters set out above.

[0255] Accordingly, the compositions, methods of treatment, or medical uses of the invention may be used to inhibit or disrupt binding of PSGL-1 to chemokines.

[0256] Inhibition or disruption of adhesion of cells

[0257] Interaction of PSGL-1 with its ligands contributes to increased cell adhesion through a number of different mechanisms. Attachment of PSGL-1 on white blood cells to ligands on the blood vessel wall is known to be an essential step in the adhesion of cells to substrates such as the endothelial lining of the circulation or platelets. Interaction of PSGL-1 on cells such as eosinophils with P-selectin on the surface of activated platelets also leads to elevated expression of adhesion molecules by the leukocytes, which increases their tendency to adhere. In either case, PSGL-1 -mediated adhesion of leukocytes, such as eosinophils, may then be further followed by other activities associated with cell migration or extravasation.

[0258] As set out in the Examples, the inventors have demonstrated that SelK2 is able to inhibit or disrupt binding of PSGL-1 to its ligands that is associated with cell adhesion. Suitably, an antibody fragment or variant to be incorporated in a pharmaceutical composition of the invention may share this ability to inhibit or disrupt binding of PSGL-1 to its ligands that is associated with cell adhesion. Inhibition or disruption of such binding will prevent or reduce cell adhesion, thus preventing or reducing biological activities that occur as a result of such cell adhesions.

[0259] Antibodies, or fragments or variants of antibodies, suitable for incorporation in the pharmaceutical compositions of the invention may be able to inhibit or disrupt PSGL-1- mediated cell adhesion by at least 10%, at least 11 %, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least 21 %, at least 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, at least 28%, at least 29%, at least 30%, at least 31 %, at least 32%, at least 33%, at least 34%, at least 35%, at least 36%, at least 37%, at least 38%, at least 39%, at least 40%, at least 41 %, at least 42%, at least 43%, at least 44%, at least 45%, at least 46%, at least 47%, at least 48%, at least 49%, at least 50%, at least 51 %, at least 52%, at least 53%, at least 54%, at least 55%, at least 56%, at least 57%, at least 58%, at least 59%, at least 60%, at least 61 %, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71 %, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or even by 100%.

[0260] The cells may be leukocytes. The cells may be epithelial cells. The cells may be selected from the group consisting of: eosinophils; neutrophils; and epithelial cells. The biological activities that will be prevented or reduced by inhibition or disruption of cell adhesion may be determined with reference to the cell type in question.

[0261] The ability of an antibody, or fragment or variant thereof, incorporated in a pharmaceutical composition of the invention to inhibit or disrupt cell adhesion mediated by binding of PSGL-1 may be investigated by any appropriate means. For instance, this may be assessed by assays of the sort discussed in Examples 2 or 4.

[0262] A pharmaceutical composition of the invention may have the ability to inhibit or disrupt adhesion of cells in accordance with any of the parameters set out above.

[0263] Accordingly, the compositions, methods of treatment, or medical uses of the invention may be used to inhibit or disrupt adhesion of cells.

[0264] Inhibition or disruption of cell rolling

[0265] Interaction of PSGL-1 with its ligands is known to contribute to the process of cell rolling exhibited by cells in the early stages of adhesion to substrates such as the endothelial lining of the circulation. This rolling may be associated with activation, and / or with subsequent migration or extravasation, of the rolling cells.

[0266] As set out in the Examples, the inventors have demonstrated that SelK2 is able to inhibit or disrupt rolling of cells mediated by binding of PSGL-1 to its ligands. Suitably, an antibody fragment or variant to be incorporated in a pharmaceutical composition of the invention may share this ability to inhibit or disrupt rolling of cells mediated by binding of PSGL-1 to its ligands. Inhibition or disruption of such cell rolling will prevent or reduce “down-stream” activities, such as cell extravasation.

[0267] As before, the cells rolling of which is to be inhibited or disrupted may be leukocytes. The cells may be non-leukocytic cells, such metastatic cancer cells. The cells may be selected from the group consisting of: neutrophils; eosinophils; and metastatic cells. The biological activities that will be prevented or reduced by the inhibition or disruption of cell rolling may be determined with reference to the cell type in question.

[0268] Antibodies, or fragments or variants of antibodies, suitable for incorporation in the pharmaceutical compositions of the invention may be able to inhibit or disrupt PSGL-1- mediated cell rolling by at least 10%, at least 11 %, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least 21 %, at least 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, at least 28%, at least 29%, at least 30%, at least 31 %, at least 32%, at least 33%, at least 34%, at least 35%, at least 36%, at least 37%, at least 38%, at least 39%, at least 40%, at least 41 %, at least 42%, at least 43%, at least 44%, at least 45%, at least 46%, at least 47%, at least 48%, at least 49%, at least 50%, at least 51 %, at least 52%, at least 53%, at least 54%, at least 55%, at least 56%, at least 57%, at least 58%, at least 59%, at least 60%, at least 61 %, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71 %, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or even by 100%.

[0269] The ability of an antibody, or fragment or variant thereof, incorporated in a pharmaceutical composition of the invention to inhibit or disrupt cell adhesion mediated by binding of PSGL-1 may be investigated by any appropriate means. For instance, this may be assessed by assays such as the neutrophil rolling assay described in Example 2.

[0270] A pharmaceutical composition of the invention may have the ability to inhibit or disrupt cell rolling in accordance with any of the parameters set out above.

[0271] Accordingly, the compositions, methods of treatment, or medical uses of the invention may be used inhibit or disrupt cell adhesion mediated by binding of PSGL-1 . Inhibition or disruption of cell extravasation

[0272] Binding of cells, such as to the endothelial lining of the circulation system, via the interaction of PSGL-1 with its ligands, is a necessary step in the process of cell extravasation. Extravasated cells contribute to inflammatory responses within tissues that can have deleterious effects upon the tissue in question.

[0273] As set out in the Examples, the inventors have demonstrated that SelK2 is able to inhibit or disrupt cell extravasation mediated by binding of PSGL-1 to its ligands. Suitably, an antibody fragment or variant to be incorporated in a pharmaceutical composition of the invention may share this ability inhibit or disrupt cell extravasation mediated by binding of PSGL-1 to its ligands. Inhibition or disruption of such cell extravasation is thus able to prevent or reduce inflammation that is mediated by the extravasated cells.

[0274] The extravasation inhibited or disrupted may be leukocyte extravasation, or may be extravasation of other cells, such as metastatic cancer cells. The cells may be selected from the group consisting of: eosinophils; neutrophils; and metastatic cancer cells. The biological activities that will be prevented or reduced by the inhibition or disruption of cell rolling may be determined with reference to the cell type in question. In the case of leukocytes, this will typically bring about a reduction in inflammation, in the case of metastatic cells it may bring about a reduction in cancer metastasis.

[0275] Antibodies, or fragments or variants of antibodies, suitable for incorporation in the pharmaceutical compositions of the invention may be able to inhibit or disrupt cell extravasation by at least 10%, at least 11 %, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least 21 %, at least 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, at least 28%, at least 29%, at least 30%, at least 31 %, at least 32%, at least 33%, at least 34%, at least 35%, at least 36%, at least 37%, at least 38%, at least 39%, at least 40%, at least 41 %, at least 42%, at least 43%, at least 44%, at least 45%, at least 46%, at least 47%, at least 48%, at least 49%, at least 50%, at least 51 %, at least 52%, at least 53%, at least 54%, at least 55%, at least 56%, at least 57%, at least 58%, at least 59%, at least 60%, at least 61 %, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71 %, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or even by 100%.

[0276] The ability of SelK2 to Inhibit or disrupt cell extravasation mediated by binding of PSGL-1 may be investigated by any appropriate means. For instance, this may be assessed by assays such as those described in Example 6.

[0277] Importantly, the inventors’ results demonstrate that a reduction in extravasation of leukocytes (such as eosinophils or neutrophils) on treatment with an antibody of the invention is not associated with a reduction in the number of the leukocytes present in the circulation. This indicates that the reduction in extravasation is not simply a function of reduced circulating leukocyte numbers, but is specifically associated with a reduction in the numbers of leukocytes (such as eosinophils or neutrophils) extravasating from the blood stream into the underlying tissues during the inflammatory response.

[0278] That circulating leukocyte numbers are maintained is important, as these cells have important roles in implementing appropriate, rather than aberrant, immune responses. Merely by way of example, circulating eosinophils play an important role in the body’s response to parasites, and neutrophils have a key role in the innate immune response.

[0279] Accordingly, the pharmaceutical compositions of the invention may be used to therapeutically reduce numbers of extravasated leukocytes (such as eosinophils or neutrophils) without adversely impacting a recipient’s immunity.

[0280] A pharmaceutical composition of the invention may have the ability to inhibit or disrupt cell extravasation in accordance with any of the parameters set out above.

[0281] Reduction of cell numbers in sputum

[0282] Pharmaceutical compositions of the invention, and the methods of treatment and medical uses of the invention, are of particular interest in the treatment of respiratory conditions or diseases, such as asthma or COPD.

[0283] Respiratory conditions or diseases may be associated with the accumulation of cells within the sputum. This accumulation may arise as a result of cell extravasation (for example, in the case of leukocytes) or as a result of shedding of cells (for example in the case of epithelial cells lining the respiratory tract). Accordingly, the ability of a SelK2 to bring about a reduction in the number of cells in the sputum may provide an indication that a pharmaceutical composition comprising this antibody is alleviating the respiratory condition in a manner suitable achieve a therapeutic effect. Suitably, an antibody fragment or variant to be incorporated in a pharmaceutical composition of the invention may share this ability to reduce the number of cells in sputum. The reduction may be noted in respect of any suitable cells of interest, such as those selected from the group consisting of: eosinophils; neutrophils; and epithelial cells.

[0284] The inventors have found that SelK2 is able to reduce the number of cells accumulating in sputum. It will be appreciated that closely related antibodies, fragments, or variants, of the sort considered in this specification, may also achieve a similar reduction in cell numbers. This reduction may provide a useful proxy for assessment of disease severity or progression.

[0285] Antibodies, or fragments or variants of antibodies, suitable for incorporation in the pharmaceutical compositions of the invention may be able to reduce cell numbers in sputum by at least 10%, at least 11 %, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least 21 %, at least 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, at least 28%, at least 29%, at least 30%, at least 31 %, at least 32%, at least 33%, at least 34%, at least 35%, at least 36%, at least 37%, at least 38%, at least 39%, at least 40%, at least 41 %, at least 42%, at least 43%, at least 44%, at least 45%, at least 46%, at least 47%, at least 48%, at least 49%, at least 50%, at least 51 %, at least 52%, at least 53%, at least 54%, at least 55%, at least 56%, at least 57%, at least 58%, at least 59%, at least 60%, at least 61 %, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71 %, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or even by 100%.

[0286] The ability of an antibody, or fragment or variant thereof, incorporated in a pharmaceutical composition of the invention to inhibit or disrupt cell extravasation mediated by binding of PSGL-1 may be investigated by any appropriate means. For instance, this may be assessed by assays such as the absolute differential eosinophil count or percentage differential eosinophil count studies described in Example 6, or the absolute differential eosinophil count or percentage differential eosinophil count studies described in Example 9. A pharmaceutical composition of the invention may have the ability to bring about a reduction in cell numbers in sputum in accordance with any of the parameters set out above.

[0287] Accordingly, the compositions, methods of treatment, or medical uses of the invention may be used to reduce the cell numbers in sputum.

[0288] Improvement in pulmonary function test

[0289] Perhaps the most important finding from the inventors’ studies undertaken in human subjects as part of a clinical trial is that SelK2 is able to bring about an improvement in lung function as demonstrated by improvement in a pulmonary function test. Suitably, an antibody fragment or variant to be incorporated in a pharmaceutical composition of the invention may also share this ability.

[0290] In particular, the inventors have demonstrated that asthma patients receiving treatment using SelK2 achieve a dramatic and statistically significant improvement in forced expiratory volume (FEV1 ). FEV1 provides a measurement of the maximum volume of air that an individual can forcefully exhale in one second. A higher FEV1 value indicates better lung function, and a lower FEV1 value is indicative of poorer lung function.

[0291] A subject or patient’s FEV1 may be measured at baseline (before treatment with an experimental agent or placebo) and after treatment. These values may then be compared to allow the calculation of a percentage change in FEV1 (for example the maximum fall in FEV1) or to allow calculation of area under the curve (AUC) in respect of FEV1 values obtained. Either of these statistical values may be useful in determining the effect of an antibody on improvement of pulmonary function.

[0292] As set out in Example 6, asthma patients challenged with an appropriate allergen exhibit a fall in FEV1 , associated with obstruction of the airways. Even when administered at a lower concentration than can be achieved with the pharmaceutical compositions of the invention, treatment with SelK2 results in a reduction in the fall in FEV1 during the late allergen response (LAR; 3 to 8 hours following allergen challenge). This indicates that lung function in those asthma patients receiving treatment with SelK2 is improved as compared to control patients receiving placebo. This is observed in relation to both the area under the curve for % FEV1 fall, and the maximum % fall in FEV1 . The change in respect of maximum % fall indicates a statistically significant improvement in lung function in subjects receiving treatment with the antibody of the invention. It will be appreciated that the pharmaceutical compositions of the invention, able to provide SelK2 (or closely related antibody fragments or variants) at much higher concentrations, may be expected to achieve improvements in lung function of the same magnitude, if not greater.

[0293] Antibodies, or fragments or variants of antibodies, suitable for incorporation in the pharmaceutical compositions of the invention may be able to improve lung function by at least 10%, at least 11 %, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least 21 %, at least 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, at least 28%, at least 29%, at least 30%, at least 31 %, at least 32%, at least 33%, at least 34%, at least 35%, at least 36%, at least 37%, at least 38%, at least 39%, at least 40%, at least 41 %, at least 42%, at least 43%, at least 44%, at least 45%, at least 46%, at least 47%, at least 48%, at least 49%, at least 50%, at least 51 %, at least 52%, at least 53%, at least 54%, at least 55%, at least 56%, at least 57%, at least 58%, at least 59%, at least 60%, at least 61 %, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71 %, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or even by 100%.

[0294] The Examples demonstrated that compositions comprising SelK2 at lower concentrations than those provided by the pharmaceutical compositions of the invention are able to improve lung function in asthmatic patients by at least 35% as compared to placebo. Accordingly, pharmaceutical compositions of the invention may improve lung function in asthmatic patients by at 35% or by at least 50%, or more, as compared to placebo. These changes are greater than those that can be achieved using currently available therapies, and so demonstrate the improvements in treatment that are made available to patients with lung diseases by the development of the pharmaceutical compositions disclosed herein.

[0295] Suitably the improvement in lung function may be demonstrated in an asthma patient undergoing allergen challenge with respect to an improvement in the maximum % fall in FEV1 as compared to placebo control. Suitably such an improvement may occur during the LAR. A pharmaceutical composition of the invention or compositions suitable for use in methods of treatment or medical uses of the invention may have the ability to bring about an improvement in a pulmonary function test in accordance with any of the parameters set out above.

[0296] Accordingly, the compositions, methods of treatment, or medical uses of the invention may be used to bring about an improvement in a pulmonary function test.

[0297] Antagonistic effects of pharmaceutical compositions of the invention on PSGL-1 signalling

[0298] Inhibition or disruption of binding of PSGL-1 to its ligands and reduction of T-cell exhaustion

[0299] Reversal of T-cell exhaustion is linked to treatment of infection, e.g. chronic infection, and cancer.

[0300] As discussed elsewhere in the specification, T-cell exhaustion is characterised by progressive loss of T-cell responsiveness, elimination of T-cells by apoptosis, and functional impairment and arrest of surviving T-cells. A loss of T-cell responsiveness in the context of infection, e.g. chronic infection, or cancer can thwart effective immune control. The mechanisms of T-cell exhaustion are incompletely understood, though it is often associated with an upregulation of immune co-inhibitory receptors (such as PD-1 , LAG3, CD160 and BTLA), loss of motility, altered transcriptional regulation, and expression of inhibitory cytokines (such as IL-10 and TGFβ). Reversal of T-cell exhaustion can therefore be linked to disease resolution against infection, e.g. chronic infection, and cancer.

[0301] As discussed elsewhere in the specification, PSGL-1 is highly expressed on T-cells following infection, e.g. chronic infection, and knockout of PSGL-1 (Selplg- / -) in a chronic infection model has been found to result in increased T-cell accumulation, preservation of CD4+ and CD8+ T-cell effectors, production of pro-inflammatory cytokines, and diminished expression of several inhibitory receptors. Similarly, knockout of PSGL-1 prevents T-cell exhaustion and reduction of tumour growth in an aggressive melanoma model. Thus, PSGL-1 has been identified as a potent negative regulator of T-cell function in infection, e.g. chronic infection, and cancer.

[0302] The ability of an antibody to reduce T-cell exhaustion may be demonstrated by T-cell response and effector function being maintained by T-cells in the presence of the antibody. Accordingly, an antibody or fragment (or an amount of such an antibody or fragment) shown to be able to maintain T-cell response and effector function in conditions that would otherwise result in a reduction of such T-cell response and / or effectorfunction should be considered an antibody or fragment (or an amount of such an antibody or fragment) that is able to reduce T-cell exhaustion. Suitable assays by which this may be assessed will be known to those skilled in the art. By way of example, Example 14 demonstrates that T-cells cultured in the presence of the exemplary antibody SelK2 do not exhibit any loss of T-cell response or effector function, assessed with reference to their production of the effector molecule interferon-γ. This is consistent with an ability of SelK2 to reduce T-cell exhaustion. In contrast, other anti-PSGL-1 antibodies have been reported to reduce T-cell responses, demonstrating their tendency to cause, or contribute to, T-cell exhaustion. Such antibodies have been reported to exert an agonistic effect on PSGL-1 signalling, in contrast to the antagonistic effects referred to in the present disclosure.

[0303] VISTA (V-domain immunoglobulin suppressor of T-cell activation; also known as B7-H5, PD- 1 H, Gi24, Diesl , SISP1 and DD1α) is a B7 family ligand that is expressed on circulating and intratumoural myeloid cellsand weakly expressed on activated lymphocytes. The extracellular domain of VISTA is rich in histidine residues, of which the imidazole sidechain of histidine protonates at physiologically relevant pH. It has been found that VISTA preferentially engages and suppresses T-cells selectively in acidic environments such as tumour microenvironments, where pH values as low as 5.85 have been measured. It has been found that VISTA binds PSGL-1 selectively at acidic pH via the extracellular histidine residues. Thus, VISTA is an acidic pH-selective ligand for PSGL-1 , and, without being bound by theory, PSGL-1 functions as a checkpoint regulator.

[0304] Antibodies engineered to selectively bind and block the VISTA-PSGL-1 interaction in acidic environments (e.g., tumour microenvironments), e.g., by binding PSGL-1 , may therefore reverse VISTA-mediated immune suppression in vivo. Accordingly, antibodies targeting PSGL-1 may reduce T-cell exhaustion.

[0305] The parental antibody from which the humanised anti-PSGL-1 antibodies for use in the methods of treatment and medical uses of the invention are derived has been found to inhibit VISTA binding to PSGL-1 in acidic environments (pH=6.0). As described further elsewhere in this specification, the inventors have demonstrated that the exemplary humanised antibody SelK2 (consisting of SEQ ID NO: 1 and SEQ ID NO: 2), has higher affinity for PSGL-1 than the parental antibody. Thus, it is expected that pharmaceutical compositions of the invention comprising SelK2 (or closely related antibody fragments or variants) will also share the capacity to block the VISTA-PSGL-1 interaction in acidic environments (pH <7).

[0306] Accordingly, in a suitable embodiment, a pharmaceutical composition of the invention may be used as a medicament for the treatment of a disease or condition by decreasing T-cell exhaustion (e.g., by antagonising PSGL-1 activity). In a suitable embodiment, the pharmaceutical compositions of the invention are suitable for treating cancer (e.g. by decreasing T-cell exhaustion). In a suitable embodiment, the pharmaceutical compositions disclosed herein are suitable for treating infection, e.g., chronic infection (e.g., by decreasing T-cell exhaustion).

[0307] Assays for measuring a reduction in T-cell exhaustion will be known to the skilled person. For instance, expression of immune checkpoint inhibitors such as PD-1 , LAG-3, and TIM-3 (which, in some embodiments, are increased in T-cell exhaustion) can be detected, as well as effector cytokines, such as IFNγ and TNFα (which, in some embodiments, are decreased in T-cell exhaustion), may also be quantified, e.g., using a sandwich immunoassay format.

[0308] In some embodiments, pharmaceutical compositions of the invention may be able to reduce T-cell exhaustion, by at least 10%, at least 11 %, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least 21 %, at least 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, at least 28%, at least 29%, at least 30%, at least 31 %, at least 32%, at least 33%, at least 34%, at least 35%, at least 36%, at least 37%, at least 38%, at least 39%, at least 40%, at least 41 %, at least 42%, at least 43%, at least 44%, at least 45%, at least 46%, at least 47%, at least 48%, at least 49%, at least 50%, at least 51 %, at least 52%, at least 53%, at least 54%, at least 55%, at least 56%, at least 57%, at least 58%, at least 59%, at least 60%, at least 61 %, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71 %, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or even by 100%, relative to a reference control.

[0309] In some embodiments, pharmaceutical compositions of the invention, may be able to increase expression of effector cytokines, e.g., IFNγ and / or TNFα, by at least 10%, at least 11 %, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least 21 %, at least 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, at least 28%, at least 29%, at least 30%, at least 31 %, at least 32%, at least 33%, at least 34%, at least 35%, at least 36%, at least 37%, at least 38%, at least 39%, at least 40%, at least 41 %, at least 42%, at least 43%, at least 44%, at least 45%, at least 46%, at least 47%, at least 48%, at least 49%, at least 50%, at least 51 %, at least 52%, at least 53%, at least 54%, at least 55%, at least 56%, at least 57%, at least 58%, at least 59%, at least 60%, at least 61 %, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71 %, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or even by 100%, relative to a reference control.

[0310] In some embodiments, pharmaceutical compositions of the invention may be able to reduce expression ofimmune checkpointinhibitors, e.g., PD-1 , LAG-3, and / or TIM-3, by at least 10%, at least 11 %, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least 21 %, at least 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, at least 28%, at least 29%, at least 30%, at least 31 %, at least 32%, at least 33%, at least 34%, at least 35%, at least 36%, at least 37%, at least 38%, at least 39%, at least 40%, at least 41 %, at least 42%, at least 43%, at least 44%, at least 45%, at least 46%, at least 47%, at least 48%, at least 49%, at least 50%, at least 51 %, at least 52%, at least 53%, at least 54%, at least 55%, at least 56%, at least 57%, at least 58%, at least 59%, at least 60%, at least 61 %, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71 %, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or even by 100%, relative to a reference control.

[0311] While PSGL-1 binding to selectins is known to mediate extravasation, it is believed that effector immune cells are still able to transmigrate into a cancer tumour bed even if extravasation via PSGL-1 / P-selectin interaction is blocked. Without being bound by theory, it is believed that VISTA expression in tumour microenvironments is high and P-selectin expression is low. Thus, the VISTA-PSGL-1 binding may be the dominant interaction, and additional PSGL-1 binding partners may also contribute to PSGL-1 -dependent inhibition in the tumour draining lymph node and / or the tumour microenvironment.

[0312] In a suitable embodiment, pharmaceutical compositions of the invention are suitable for use as a medicament for the treatment of a disease or condition by preventing T-cell exhaustion, e.g., in a subject suffering from cancer or infection, e.g., chronic infection.

[0313] In the context of the present disclosure, references to the “prevention” ofa disease or condition associated with PSGL-1 (e.g., T-cell exhaustion) may be taken as referring to prophylactic medical intervention that is intended to stop one or more incidences of a disease or condition associated with PSGL-1 signalling (e.g., T-cell exhaustion) from occurring or from further developing or progressing. Pharmaceutical compositions of the invention may be used to prevent a disease or condition by inhibiting the formation of new binding between PSGL-1 and one or more of its ligands and / or antagonising PSGL-1 activity.

[0314] In the present context, prevention may result in the complete avoidance of symptoms of a disease or condition (for example such that T-cell exhaustion does not occur following onset of cancer or infection, e.g., chronic infection), or partial avoidance of symptoms of a disease or condition (for example, such that T-cell exhaustion is reduced or delayed).

[0315] In methods of treatment or medical uses in accordance with such embodiments of the invention, a concentration of an antibody incorporated in a pharmaceutical composition of the invention may be maintained in the circulation that is sufficient to achieve at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80% or at least 90% inhibition of PSGL- 1 activity. This approach may thus reduce T-cell exhaustion that contributes to a disease or condition (such as persistence of cancerorinfection, e.g., chronic infection). Alternatively, this approach may prevent the onset or development of T-cell exhaustion that may otherwise contribute to a disease or condition (such as persistence of cancer or infection, e.g., chronic infection). Suitable routes of administration and dosing regimens may be selected with this aim in mind.

[0316] Pharmaceutical compositions of the invention may comprise antibodies able to inhibit and / or disrupt PSGL-1 activity. In the context of the present invention, “inhibition” of binding of PSGL- 1 to a ligand may be taken as referring to the prevention of the formation of new interactions between PSGL-1 and the ligand in question. On the other hand, “disruption” of binding of PSGL-1 to a ligand may be taken as referring to the breaking down of previously formed interactions between PSGL-1 and the ligand in question. Pharmaceutical compositions of the invention may inhibit or disrupt binding of PSGL-1 to a ligand such as VISTA, by at least 10%, at least 11 %, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least 21 %, at least 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, at least 28%, at least 29%, at least 30%, at least 31 %, at least 32%, at least 33%, at least 34%, at least 35%, at least 36%, at least 37%, at least 38%, at least 39%, at least 40%, at least 41 %, at least 42%, at least 43%, at least 44%, at least 45%, at least 46%, at least 47%, at least 48%, at least 49%, at least 50%, at least 51 %, at least 52%, at least 53%, at least 54%, at least 55%, at least 56%, at least 57%, at least 58%, at least 59%, at least 60%, at least 61 %, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71 %, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or even by 100%.

[0317] The skilled person will readily be able to identify suitable assays for determining an extent of inhibition ordisruption of binding achieved. These may be selected with reference to particular ligands of interest (for example, binding of PSGL-1 to VISTA), or with reference to particular binding-associated activities of interest (for example, T-cell activity).

[0318] While the ability of pharmaceutical compositions of the invention to block the VISTA-PSGL-1 interaction makes them particularly suitable for practicing the methods of treatment or the medical uses of the invention, it will be recognised that, in an alternative embodiment, these methods or uses may also potentially be practiced using anti-PSGL-1 antibodies disclosed herein to affect biological systems independently of the VISTA-PSGL-1 pathway.

[0319] A suitable pharmaceutical composition of the invention may have the ability to inhibitor disrupt binding of PSGL-1 to its ligands in accordance with any of the parameters set out above.

[0320] Inhibition or disruption of PSGL-1 signalling and reduction of T-cell exhaustion

[0321] As discussed elsewhere in the disclosure, PSGL-1 has been identified as a potent negative regulator of T-cell function in infection, e.g. chronic infection, and cancer. PSGL-1 functions as a cell signalling transmembrane receptor. The antibodies for use in the methods of treatment and medical uses of the invention may, in some embodiments, antagonise PSGL-1 activity. Without being bound by theory, antagonising the PSGL-1 signalling cascade, e.g., in T-cells, can reverse T-cell exhaustion.

[0322] As used herein, referenceto “antagonists” and “antagonising” in the context of PSGL-1 activity relate to reducing or preventing the activation of the PSGL-1 receptor and thereby preventing or inhibiting a biological response arisingfrom downstream intracellular signalling associated with said receptor. The receptor may be present on a leukocyte, for instance, a T-cell. Thus, antagonists may have affinity but no efficacy for their cognate receptors, and binding may disrupt the interaction and inhibit the function of an agonist or inverse agonist at the receptor. Typically, antagonists mediate their effects by binding to the active site or to the allosteric site on a receptor, or they may interact at unique binding sites not normally involved in the biological regulation of the receptor's activity. Thus, in some embodiments, pharmaceutical compositions of the invention incorporate an antibody which directly binds and blocks the receptor. In other embodiments, pharmaceutical compositions of the invention incorporate an antibody which binds to a different binding site that indirectly results in the inactivation of the receptor.

[0323] Much of what is known about signalling mechanismsthat can be engaged by PSGL-1 comes from studies of neutrophils.

[0324] Src family kinases (SFKs) are non-receptor tyrosine kinases involved in the regulation of cellular functions such as cell proliferation, differentiation, apoptosis, migration, and metabolism. The vertebrate Src kinase family is composed of nine members, namely, SRC, LCK, LYN, BLK, HCK, FYN, FGR, YES, and YRK (YRK is only found in chickens). Src Family Kinases are understood to be an element of the downstream signalling pathway of PSGL-1 , and are understood to play a role in promoting phosphorylation of downstream effectors in an ITAM-dependent pathway.

[0325] Spleen tyrosine kinase (SYK) is also known to relay immune receptor signalling. The Syk- ZAP70 family has only two members and only Syk is found in innate cells. SYK is a 72 kDa non-receptor tyrosine kinase, which contains two SRC homology 2 (SH2)-domains and a kinase domain, and is most highly expressed by haematopoietic cells. Mammals also express a SYK homologue, ZAP70, which is mostly restricted to T- and NK-lineage cells. Syk kinase is required for initiation of signalling involving receptors that utilize immunoreceptortyrosine activation (ITAM) domains. Src-family and Syk kinases can operate alone or together to activate inhibitory signalling pathways.

[0326] During signalling, PSGL-1 and other adhesion receptors such as CD44 move to the uropod through cytoskeletal rearrangement that depends on interactions with the ERM (Ezrin, Radixin, Moesin) proteins ezrin and moesin. Ezrin and moesin are proteins which anchor PSGL-1 to the actin cytoskeleton and act as adaptors that facilitate intracellular signalling.

[0327] Extracellular signal-regulated kinase 1 / 2 (ERK) belongs to the mitogen-activated protein kinase (MAPK) family, which plays a role in signalling cascades and transmits extracellular signals to intracellular targets. Activation of PSGL-1 on neutrophils by antibody ligation is understood to induce the phosphorylation of multiple proteins including extracellular-signal related kinase (ERK)1 and ERK2.

[0328] The main Syk family kinase in T-cells is understood to be Zap70. It is not known whether Zap70 interacts with the ITAM-like sequence in ERM proteins and lead to signal transduction in T-cells downstream of PSGL-1. For T-cell receptor (TCR) signalling, ezrin is needed for proper Zap70 recruitment to the immune synapse, although after initial recruitment into the synapse, ezrin and moesin move to the uropod of the cell. PSGL-1 also becomes excluded from the immune synapse on T-cells, and then localizes to the uropod. It is understood that TCR signalling may be reduced via the downstream ERKandAKT pathways when PSGL-1 is cross-linked with the antibody at the time of TCR stimulation. While it is possible that crosslinking PSGL-1 prevents optimal formation of the synapse, the precise localization of PSGL-1 in T-cells is not fully understood.

[0329] Without being bound by theory, it is understood that activation of PSGL-1 paired with TCR stimulation results in early dephosphorylation of the ERK and AKT (Protein Kinase B) pathways, indicating that PSGL-1 signalling can extinguish TCR signalling events (Tinoco et al. 2017 Trends Immunol. 2017 May; 38(5): 323-335). However, the precise nature of how PSGL-1 signalling contributes to this is unknown. Thus, without being bound by theory, antagonism of PSGL-1 signalling may reverse the inhibition of TCR signalling and thereby reduce T-cell exhaustion.

[0330] The parental antibody from which SelK2 (the antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2) is derived has been found not to activate the Src Family of Kinases in a neutrophil study. In contrast, membrane P-selectin in solution or immobilized does activate SFKs (most likely because oligomeric P-selectin can crosslink PSGL-1 ).

[0331] The present inventors have demonstrated that SelK2, has h ig her affin ity for PSGL-1 than their parental antibody. Thus, it is expected that pharmaceutical compositions of the invention (which comprise SelK2 or closely related antibody fragments or variants) will similarly not activate Src Family of Kinases signalling. Accordingly, the pharmaceutical compositions of the invention for use in the methods of treatment or medical uses of the invention may surprisingly not induce PSGL-1 downstream intracellular signalling despite comprising antibodies, antibody fragments or variants that bind PSGL-1 with high affinity.

[0332] Accordingly, provided herein is a pharmaceutical composition of the invention for use as a medicament for the treatment of a disease or condition by antagonising PSGL-1 activity. Also provided herein is a method of treating a disease or condition comprising administering to a subject in need thereof a pharmaceutical composition of the invention in an amount sufficient to antagonise PSGL-1 activity.

[0333] Thus, without being bound by theory, antagonising PSGL-1 signalling, e.g., in T-cells, can reverse T-cell exhaustion. Accordingly, in an embodiment, provided herein is a pharmaceutical composition of the invention for use as a medicamentfor the treatment of a disease or condition by antagonising PSGL-1 activity and thereby decreasing T-cell exhaustion. Also provided herein is a method of treating a disease or condition comprising administering to a subject in need thereof a pharmaceutical composition of the invention in an amount sufficient to antagonise PSGL-1 activity, wherein the pharmaceutical composition is administered to the subject in an amount sufficient to reduce T-cell exhaustion.

[0334] As discussed elsewhere in the disclosure, PSGL-1 has been identified as a potent negative regulator of T-cell function in infection, e.g. chronic infection, and cancer. Thus, in a further embodiment, a pharmaceutical composition of the invention is provided for use as a medicamentforthe treatment of cancer, e.g., by antagonising PSGL-1 activity and / or reducing T-cell exhaustion. In anotherembodiment, provided herein is a pharmaceutical composition of the invention for use as a medicament forthe treatment of infection, e.g. chronic infection, e.g., by antagonising PSGL-1 activity and / or reducing T-cell exhaustion.

[0335] In some embodiments, the pharmaceutical compositions of the invention use in the methods of treatment or medical uses of the invention inhibit Src Family of Kinases signalling. In an alternative embodiment, the pharmaceutical compositions of the invention for use in the methods of treatment or medical uses of the invention activate Src Family of Kinases signalling. In another embodiment, the pharmaceutical compositions of the invention for use in the methods of treatment or medical uses of the invention inhibit the Syk Family of Kinases signalling. In an alternative embodiment, the pharmaceutical compositions of the invention for use in the methods of treatment or medical uses of the invention activate the Syk Family of Kinases signalling. In some embodiments, the pharmaceutical compositions of the invention for use in the methods of treatment or medical uses of the invention increases or activates Erk signalling. In alternative embodiments, the pharmaceutical compositions of the invention for use in the methods oftreatment or medical uses of the invention inhibit Erk signalling. In some embodiments, the pharmaceutical compositions of the invention for use in the methods of treatment or medical uses of the invention increase or activate Akt signalling. In alternative embodiments, the pharmaceutical compositions of the invention for use in the methods of treatment or medical uses of the invention inhibit Akt signalling. In another embodiment, ezrin activity is increased or activated. In an alternative embodiment, ezrin activity is inhibited. In yet another embodiment, moesin activity is increased or activated. In an alternative embodiment, moesin activity is inhibited. In another embodiment, one or more other intracellular signal pathways are inhibited or activated by a pharmaceutical composition of the invention.

[0336] The skilled person will readily be able to identify suitable assays for determining an extent of antagonism or inhibition of intracellular signalling achieved. The skilled person will recognise that there are multiple common techniques that may be used to determine the extent of antagonism or inhibition of intracellularsignalling achieved. Suitable techniques that may be used to determine the extent of antagonism or inhibition of intracellular signalling achieved include Western Blot, immunoprecipitation, glutathione S-transferase (GST) binding, radioactivate kinase activity assays, cell behaviour assays, immunohistochemistry, flow cytometry, mass cytometry, single-cell immune-sequencing, single-cell RNA sequencing, single-cell proteomics, fluorescence resonance energy transfer (FRET), and fluorescence recovery / redistribution after photobleaching (FRAP).

[0337] A suitable antigen-binding fragment of an antibody may have the ability to antagonise PSGL- 1 signalling in accordance with any of the parameters set out above.

[0338] Low immunogenicity of SelK2

[0339] It is generally recognised thattherapeuticantibodies may give rise to immune responses within their recipients. This is particularly problematic in the case of therapeutic antibodies that need to be administered for prolonged periods of time, such as in the treatment of chronicconditions or diseases.

[0340] Even humanised antibodies may trigger an immune response in this manner. Immune responses to humanised therapeutic antibody result in the production of human anti-human antibodies (HAHAs). HAHAs bind to the therapeutic antibody and may interfere with its ability to bind to its corresponding antigen (such as PSGL-1 ). Consequently, the presence of HAHAs can in some cases significantly decrease the therapeutic effectiveness of an antibody.

[0341] Accordingly, it is advantageous if therapeutic antibodies demonstrate low immunogenicity, thus decreasing the risk of HAHA formation and loss of therapeutic effect.

[0342] SelK2 has been demonstrated to have low immunogenicity in recipients. As described in Example 8, immunogenicity analysis conducted as part of a clinical trial using SelK2 showed that responses were of low levels, some of which did not repeat at later time points, and no neutralisation of the activity of the antibody of the invention was observed. Furthermore, some patients receiving placebo were also noted to demonstrate low level responses by the assay method used.

[0343] Accordingly, it will be recognised that compositions of the invention comprising SelK2 are likely to benefit from low immunogenicity that will make them well suited to therapeutic use in practice, and particularly to use in the treatment of chronic conditions or diseases. Suitably, an antibody fragment or variant to be incorporated in a pharmaceutical composition of the invention may share this low immunogenicity by virtue of its high degree of sequence identity with SelK2.

[0344] Thus, a pharmaceutical composition of the invention may have low immunogenicity in accordance with any of the parameters set out above, and such compositions may generate low immune responses in recipients in practice.

[0345] Compositions suitable for use in methods of treatment and medical uses

[0346] In its third aspect, the invention provides a method of preventing and / or treating a disease or condition in a subject in need thereof, the method comprising providing to the subject a therapeutically effective amount of a pharmaceutical composition in accordance with the present invention. In its fourth aspect, the invention provides a pharmaceutical composition in accordance with the invention for use as a medicament.

[0347] As set out above, SelK2 has properties that confer useful therapeutic utility in a wide range of applications. These are reflected in the methods of treatment and medical uses described below. The skilled person will recognise that antibody fragments and variant sharing a high degree of sequence identity with SelK2, as defined and further described elsewhere in the specification, may be expected to have similar properties, and thereby to be similarly useful in therapeutic applications.

[0348] Since the pharmaceutical compositions of the invention comprise high concentrations of antibodies (whether SelK2, or an antibody fragment or variant) they are well suited to the provision of these therapeutic agents for methods of treatment or medical uses.

[0349] Except for where the context requires otherwise, any of the pharmaceutical compositions set out in the various aspects and embodiments of this specification should be taken as suitable compositions of the invention to be employed in any of the methods of treatment or medical uses of the invention described below.

[0350] Methods of treatment and medical uses

[0351] As noted above, SelK2 has properties that make it suitable fortherapeutic use in a number of contexts. The pharmaceutical compositions of the invention comprise SelK2, or very closely related antibody fragments or variants, at high concentration. They are thus suitable to be employed in methods or medical uses that make use of these properties. More details of these are provided below.

[0352] According to the third aspect, the invention provides a method of preventing and / or treating a disease or condition in a subject in need thereof, the method comprising providing to the subject a therapeutically effective amount of a pharmaceutical composition in accordance with the invention.

[0353] In a fourth aspect of the invention, there is provided a pharmaceutical composition in accordance with the invention for use as a medicament.

[0354] The disease or condition to be prevented or treated by a method of the third aspect of the invention or a medical use of the fourth aspect of the invention may be a condition or disease associated with binding of PSGL-1 to its ligands. A method of treatment or medical use of the invention suitable for prevention ortreatment in this context may be one that inhibits or disrupts binding of PSGL-1 to its ligands.

[0355] The disease or condition to be prevented or treated by a method of the third aspect of the invention or a medical use of the fourth aspect of the invention may be a condition or disease associated with intracellular signalling activity by PSGL-1 . A method of treatment or medical use of the invention suitable for prevention or treatment in this context may be one that antagonises intracellular signalling activity by PSGL-1 .

[0356] Examples of such conditions or diseases that may benefit from prevention or treatment using the pharmaceutical compositions of the invention are considered below.

[0357] Prevention or treatment of inflammatory disease or conditions

[0358] The skilled person will be aware of a wide range inflammatory diseases or conditions which are characterised by abnormal or dysregulated inflammation. Inflammation is a fundamental biological process that involves the activation of the immune system to defend against pathogens, remove damaged cells or debris, and initiate tissue repair. However, in inflammatory diseases, this inflammatory response becomes aberrant (for example, chronic, excessive, or misdirected), leading to tissue damage, dysfunction, and the development of clinical symptoms. Such disease may be caused by a variety of biological process such as those associated binding of PSGL-1 to its ligands.

[0359] Accordingly, the pharmaceutical compositions of the invention may be employed in methods of treatment or medical uses for the prevention or treatment of an inflammatory disease or condition.

[0360] In the context of the present disclosure, references to the “prevention” of an inflammatory disease or condition may be taken as referring to prophylactic medical intervention that is intended to stop one or more incidences of an inflammatory disease or condition from occurring or from further developing or progressing.

[0361] Many inflammatory disease or conditions are associated with binding of PSGL-1 to its ligands. The skilled person will be aware of a wide range of diseases orconditions thatare associated with binding of PSGL-1 to its ligands. Such diseases or conditions may be caused by binding of PSGL-1 to one or more of its ligands. Examples of such diseases or conditions are considered in more detail over the following pages. Pharmaceutical compositions of the invention may be employed in methods of treatment or medical uses for use in the prevention and / or treatment of such diseases or conditions associated with binding of PSGL-1 to its ligands.

[0362] In the context of the present disclosure, references to the “prevention” ofa disease or condition associated with binding of PSGL-1 to its ligands may be taken as referring to prophylactic medical intervention that is intended to stop one or more incidences of a disease or condition associated with binding of PSGL-1 to its ligands from occurring or from further developing or progressing. Compositions of the invention may be used to prevent a disease or condition by inhibiting the formation of new binding between PSGL-1 and one or more of its ligands.

[0363] References within the present disclosure to “prevention” of a disease or condition, either generically or specifically, may be taken as encompassing delaying a symptom or onset of the relevant disease or condition. Furthermore, references within the present disclosure to “prevention” of a disease or condition, either generically or specifically, should also be taken as also encompassing incomplete or partial prevention of the relevant disease or condition. Thus, prevention may result in the complete avoidance of symptoms of a disease or condition (for example such that asthma symptoms do not occuron exposure to an allergen), or partial avoidance of symptoms of a disease or condition (for example, such that symptoms experienced on exposure to an allergen are reduced or delayed).

[0364] In the case of treatment of a respiratory disease or condition such as asthma or COPD, prevention of the disease or condition may be demonstrated by improved lung function as compared to the function that would be achieved in the absence of treatment. As discussed in more detail elsewhere in this specification, lung function may be assessed with reference to changes in FEV1 in response to allergen challenge achieved with or without treatment. Lung function may be improved by at least 20%, at least 30%, at least 40%, at least 50%, or more, as compared to the function that would occur in the absence of treatment.

[0365] Alternatively, or add itionally, in prevention of a respiratory disease orconditionsuch asasthma or COPD, prevention of the disease may be demonstrated by a reduction of the number of localised inflammatory cells (such as eosinophils or neutrophils) present as compared to the numbers of such cells that would be present in the absence of treatment. It has been reported that the numberof localised inflammatory cells present in the lungs of patients with respiratory diseases or conditions such as asthma or COPD correlates with disease severity. The quantity of localised inflammatory cells present in the lungs may be assessed with reference to the number of such cells present in sputum, as considered elsewhere in this specification. The number of localised inflammatory cells present in the lungs may be reduced by at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, or more, as compared to the number of such cells present in sputum in the absence of treatment.

[0366] In methods of treatment or medical uses in accordance with the third orfourth aspects of the invention, a concentration of a therapeutically effective antibody, fragment or variant may be maintained in the circulation that is sufficient to achieve at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80% or at least 90% inhibition of binding of PSGL-1 to its ligands. This approach may thus prevent the steps of leukocyte adhesion, rolling and extravasation (particularly in respect of eosinophils) that may otherwise contribute to a disease or condition (such as an asthma attack). Suitable routes of administration and dosing regimens may be selected with this aim in mind.

[0367] Without wishing to be bound by any hypothesis, pharmaceutical compositions of the invention may be used to prevent a disease or condition by inhibiting formation of binding complexes between PSGL-1 and one or more of its ligands.

[0368] References to “treatment” of inflammatory diseases or conditions may be taken as referring to medical intervention that is intended to alleviate one or more symptoms that are already occurring in respect of an inflammatory disease or condition (as contrasted with the prophylactic preventative uses described above). “Treatment’ in this context may include complete treatment, such that all symptoms are fully alleviated, or partial treatment (in which symptoms are incompletely alleviated). Partial treatment, in which symptoms are alleviated by at least 20%, at least 40%, at least 60%, or at least 80% as compared to those occurring without treatment, may be sufficient to provide valuable relief to a subject. Suitably, pharmaceutical compositions of the invention, and the methods of treatment and medical uses of the invention, may be used in the treatment of such diseases or conditions.

[0369] References to “treatment” of diseases or condition associated with binding of PSGL-1 to its ligands may be taken as referring to medical intervention that is intended to alleviate one or more symptoms that are already occurring in respect of a disease or condition associated with binding of PSGL-1 to its ligands (as contrasted with the prophylactic preventative uses described above). “Treatment” in this context may include complete treatment, such that all symptoms are fully alleviated, or partial treatment (in which symptoms are incompletely alleviated). Partial treatment, in which symptoms are alleviated by at least 20%, at least 40%, at least 60%, or at least 80% as compared to those occurring without treatment, may be sufficient to provide valuable relief to a subject.

[0370] Without wishing to be bound by any hypothesis, pharmaceutical compositions of the invention may be used to treat a disease or condition by disrupting existing binding between PSGL-1 and one or more of its ligands.

[0371] For example, in methods of treatment or medical uses in accordance with the invention, a concentration of a therapeutically effective antibody, fragment or variant may be established in the circulation that is sufficient to disrupt binding of PSGL-1 to its ligands. This approach may thus break down existing binding, associated with an ongoing disease or condition (such as an asthma attack in progress). Disrupting binding in this manner is able to reverse leukocyte adhesion and rolling (for example in respect of eosinophils) that is in progress and prevent further extravasation associated with the condition or disease. Suitable routes of administration and dosing regimens may be selected with this aim in mind.

[0372] The compositions, methods of treatment or medical uses of the invention may be of benefit in the prevention ortreatmentofa disease or condition requiring inhibition or disruption of PSGL- 1 -mediated binding between leukocytes (such as eosinophils) and endothelial cells.

[0373] The compositions, methods of treatment or medical uses of the invention may be of benefit in the prevention ortreatmentofa disease or condition requiring inhibition or disruption of PSGL- 1 -mediated binding between leukocytes (such as eosinophils) and platelets.

[0374] The compositions, methods of treatment or medical uses of the invention may be of benefit in the prevention ortreatmentofa disease or condition requiring inhibition or disruption of PSGL- 1 -mediated binding between endothelial cells and sickled erythrocytes.

[0375] It is known that intracellular signalling through PSGL-1 plays an important role in many diseases or conditions associated with PSGL-1 activity. These diseases or conditions may also be associated with inflammatory activity. The adverse effects of PSGL-Ts intracellular signalling may arise as a result of agonism of such signalling. In this case, the therapeutic activity of pharmaceutical compositions of the invention may arise from their antagonism of intracellular PSGL-1 signalling associated with the disease or condition.

[0376] Subjects or patients The methods of treatment of the invention are practiced in respect of subjects requiring the prevention ortreatmentadisease or condition. The words “subject” and “patient” may be used interchangeably in the context of the present disclosure.

[0377] Suitably, a subject or patient may require prevention or treatment in respect of a disease or condition.

[0378] Suitably, a subject or patient may require prevention ortreatmentin respect of an inflammatory disease or condition.

[0379] The subject or patient may have symptoms of a disease or condition requiring treatment, or may be identified as being at elevated risk of developing a disease or condition requiring prevention.

[0380] In a suitable embodiment, a subject or patient may be subjectto an acute disease orcondition. In this case a method or medical use of the invention may be used to treat the disease, thereby alleviating the symptoms of the acute disease or condition. Such treatments may make use of the ability of SelK2, or related antibody fragments or variants, incorporated within pharmaceutical compositions of the invention to disrupt existing binding of PSGL-1 and its ligands.

[0381] In a suitable embodiment, a subject or patient may be subject to chronic disease. In such embodiments, a method of treatment or medical use of the invention may be utilised to prevent the worsening of the chronic disease, or incidences of acute disease. Such therapeutic prevention may make use of the ability of SelK2 , or related antibody fragments or variants, incorporated within pharmaceutical compositions of the invention to inhibit new binding occurring between PSGL-1 and its ligands.

[0382] Methods of treatment and medical uses

[0383] The methods of treatment and medical uses of the invention may be employed in the prevention or treatment of a wide range of diseases or conditions. The therapeutic applications to which the pharmaceutical compositions of the invention may be put will depend upon the nature of the therapeutically effective antibodies incorporated in these compositions. Without limitation, suitable diseases or conditions may be selected on the basis of the properties that have been demonstrated in respect of SelK2, the exemplary antibody referred to herein.

[0384] Diseases or conditions to be treated

[0385] Pharmaceutical compositions of the invention may be employed in methods of treatment or medical usesforthe prevention ortreatmentof a broad range of diseases orconditions arising as a consequence of PSGL-1 binding to its ligands. These inflammatory diseases or conditions. Such conditions or disorders may include those associated with agonistic intracellular signalling via PSGL-1.

[0386] More details of specific diseases or conditions that may benefit from prevention or treatment using the pharmaceutical compositions of the invention are set out below.

[0387] Anti-inflammatory applications

[0388] The binding of PSGL-1 with its ligands, such as P-selectin and L-selectin, is associated with many biological processes which cause inflammation. Accordingly, the ability to inhibit or disrupt such binding allows modulation of these processes for clinical purposes.

[0389] PSGL-1 binding to selectins (such as P-selectin) or to chemokines (such as CCL27) is known to be associated with the homing of inflammatory cells to sites of inflammation. The ability of the SelK2 to inhibit or disrupt binding of PSGL-1 to these ligands may facilitate inhibition of the pathophysiological recruitment and transmigration of leukocytes (including eosinophils, neutrophils, and lymphocytes), such as that occurring into the tissue and airways of the lung in asthma and COPD. Thus, the results obtained by the inventors support a scientific rationale for the ability of the pharmaceutical compositions of the invention (which comprise high concentrations of SelK2 or its closely related antibody fragments or variants) to prevent these activities in a therapeutically effective manner.

[0390] In view of the common role that the binding of PSGL-1 to its ligands plays in the initiation or propagation of damaging biological activities, particularly those associated with extravasation and influx of inflammatory cells, the skilled person will also appreciate that the compositions, methods of treatment, and medical uses of the invention may also be expected to have clinical applications beyond those directly demonstrated herein. Pharmaceutical compositions of the invention, are suitable for use as anti-inflammatory agents. Suitably, pharmaceutical compositions, methods of treatment and medical uses of the invention are suitable for use in the prevention or treatment of an inflammatory disease or condition.

[0391] The pharmaceutical compositions, methods of treatment and medical uses may be of use in the prevention or treatment of inflammatory conditions or diseases selected from the group consisting of: asthma; chronic obstructive pulmonary disease (COPD); allergic reactions; inflammatory bowel disease (e.g., Crohn's disease, ulcerative colitis, enteritis); arthritis (e.g., rheumatoid arthritis, osteoarthritis, psoriatic arthritis); graft rejection; graft versus host disease; psoriasis; dermatitis; nephritis; lupus erythematosus; scleroderma; rhinitis; anaphylaxis; diabetes; multiple sclerosis; atherosclerosis; sickle cell disease (SCD); and thyroiditis.

[0392] The pharmaceutical compositions, methods of treatment and medical uses of the invention are of particular use in the prevention or treatment of inflammatory diseases or conditions associated with the extravasation and accumulation of inflammatory cell types selected from the group consisting of: eosinophils; neutrophils; and lymphocytes (such as T-cells).

[0393] Suitably, pharmaceutical compositions, methods of treatment and medical uses of the invention are of use in the prevention or treatment of an eosinophilic disease or condition. Alternatively, pharmaceutical compositions, methods of treatment and medical uses of the invention are of use in the prevention or treatment of a neutrophilic disease or condition. Suitably, pharmaceutical compositions, methods of treatment and medical uses of the invention are of use in the prevention or treatment of a lymphocytic disease or condition.

[0394] Eosinophilic conditions and diseases

[0395] Eosinophils are a type of white blood cell that contributes to initiation and modulation of inflammation. Eosinophilic conditions occur when large numbers of eosinophils are recruited to specific sites within the body. This may particularly arise as a result of extravasation, with eosinophils cross blood vessel walls to enter inflamed tissue. The presence of increased number of eosinophils within tissues and cause them to become damaged.

[0396] SelK2, as demonstrated in the Examples has been proven to be clinically effective in alleviating eosinophilic conditions. Accordingly, the use of the pharmaceutical composition of the invention, and / or the methods of treatment and medical uses of the invention, for the prevention or treatment of eosinophilic conditions constitutes a particularly suitable embodiment of these various aspects of the invention.

[0397] In a suitable embodiment, the eosinophiliccondition ordisease to be prevented ortreated may be selected from the group consisting of: eosinophilic asthma; allergies; eosinophilic oesophagitis; eosinophilic dermatitis; acute myelogenous leukemia (AML), ascariasis; atopic dermatitis (eczema); bullous pemphigoid; cancer (such as Hodgkin lymphoma, leukemia, and certain myeloproliferative neoplasms); Churg-Strauss syndrome; drug allergy; eosinophilic cardiomyopathy; eosinophilic cellulitis (Wells’ syndrome); eosinophilic colitis; eosinophilic enteritis;; eosinophilic fasciitis; eosinophilic gastrointestinal diseases; eosinophilic granulomatosis with polyangiitis (EGPA); eosinophilic leukemia; eosinophilic myocarditis; hay fever (allergic rhinitis); Hodgkin's lymphoma (Hodgkin's disease); hypereosinophilic syndromes; idiopathic hypereosinophilic syndrome (HES); lgG4-Related Disease; inflammatory bowel disease (Crohn's disease, ulcerative colitis); lymphatic filariasis; neuromyelitis optica (NMO); ovarian cancer; parasitic infection; primary biliary cirrhosis; primary immunodeficiency; and trichinosis.

[0398] Asthma

[0399] Asthma is a disease associated with long-term inflammation of the airways of the lungs. In asthma attacks constriction of the airways and bronchospasms lead to shortness of breath, tightness of the chest, and other symptoms such as coughing and wheezing. Attacks may be triggered by activities, or a range of environmental conditions, and asthma may include an allergic component. Asthma is believed to have been responsible for around 461 ,000 deaths worldwide in 2019.

[0400] In the Examples, SelK2 is shown to be effective in the prevention or treatment of asthma. Thus, pharmaceutical compositions, methods of treatment and medical uses of the invention, which employ antibodies, fragments or variants closely structurally related to SelK2, may be used effectively in the prevention or treatment of asthma. The compositions, and methods of treatment or medical uses, of the invention may be of benefit in the prevention or treatment of moderate or severe asthma. In particular, they may be useful in the prevention or treatment of eosinophilic asthma.

[0401] As the Examples clearly demonstrate, treatment of asthma using SelK2, is able to bring about a statistically significant improvement in lung function. Pharmaceutical compositions of the invention thus represents an effective asthma treatment agent, and the methods of treatment and medical uses of the invention thus represent an effective asthma therapy. Indeed, the results show that the improvement provided by SelK2 is greater than that achieved using leading treatments known from the prior art, and so represents an important advance in the therapies available to asthma patients. The high concentrations of SelK2 (or closely structurally related fragments or variants) incorporated in the pharmaceutical compositions of the invention may be expected to be even more effective, and to achieve greater therapeutic impact than leading treatments known from the prior art, or the pharmaceutical formulations used in the Examples.

[0402] The ability of SelK2 to inhibit or disrupt binding of PSGL-1 to its ligands indicates that the pharmaceutical compositions of the invention may suitably be used in asthma management and therapy to prevent the occurrence of asthma attacks, or to alleviate symptoms associated with ongoing attacks.

[0403] In particular, the methods of treatment or medical uses of the invention may be used for the prevention or treatment of asthma by attenuation of allergen-induced bronchioconstriction. The data disclosed in the Examples indicate that SelK2 is effective in reducing such constriction and thus pharmaceutical compositions of the invention comprising high concentrations of SelK2 or closely related antibody fragments or variants may be useful in reducing such constriction.

[0404] Suitably, asthma to be prevented or treated may be selected from the group consisting of: eosinophilic asthma; allergic asthma; and severe neutrophilic asthma.

[0405] Since the compositions of the invention and / or methods of treatment and medical uses of the invention, comprise SelK2, or closely related antibody fragments or variants of this antibody which is able to bring about a reduction in the extravasation of eosinophils that drives eosinophilic asthma, they may be of particular utility in the prevention or treatment of eosinophilic asthma.

[0406] A suitable embodiment of the third aspect of the invention, provides a method of preventing or treating asthma in a subject in need thereof, the method comprising providing a therapeutically effective amount of a pharmaceutical composition of the invention to the subject.

[0407] A suitable embodiment of the fourth aspect of the invention provides a pharmaceutical composition in accordance with the invention for use in the prevention or treatment of asthma Neutrophilic diseases and disorders

[0408] Neutrophils are very abundant white blood cells that contribute to initiation and modulation of inflammation. Neutrophilic conditions occur when large numbers of neutrophils are recruited to specific sites within the body. This may particularly arise as a result of extravasation, with neutrophils cross blood vessel walls to enter inflamed tissue. The presence of increased number of neutrophils within tissues and cause them to become damaged.

[0409] As demonstrated in the Examples, SelK2 has proven to be surprisingly clinically effective in reducing the extravasation and accumulation of neutrophils. Thus, pharmaceutical compositions of the invention comprising high concentrations of SelK2, or closely related antibody fragments or variants thereof, offer promise in alleviating neutrophilic conditions. Accordingly, the use of pharmaceutical compositions of the invention, and / or the methods of treatment and medical uses of the invention, for the prevention or treatment of neutrophilic conditions constitutes a particularly suitable embodiment of these various aspects of the invention.

[0410] Thus, in a suitable embodiment, use of pharmaceutical compositions of the invention, and / or the methods of treatment and medical uses of the invention is for the prevention or treatment of neutrophilic conditions.

[0411] Suitably, the neutrophilic condition or disease to be prevented or treated may be selected from the group consisting of: COPD; neutrophilic dermatoses (e.g., Sweet's syndrome, amicrobial pustulosis of the folds, erythema elevatum diutinum, amicrobial pustulosis of the scalp / leg, amicrobial subcorneal pustulosis and pyoderma gangrenosum); neutrophilic asthma; rheumatoid arthritis (RA); cystic fibrosis (CF); neutrophilic vasculitis (e.g., antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis); gout; IBD (e.g., Crohn’s disease and ulcerative colitis); neutrophilic meningitis; behget’s disease; PAPA syndrome; hidradenitis suppurativa; acne due to EGFRIs; PASH syndrome; PAPASH syndrome, neutrophilic panniculitis; aseptic abscess syndrome; systemic inflammatory response syndrome (SIRS); severe septicaemia; cryopyrin-associated periodic syndromes (CAPS); neutrophilic otitis externa; allergies; Chronic neutrophilic leukemia (CNL); hay fever (allergic rhinitis); Severe Acute Respiratory Syndrome (SARS); Middle East Respiratory Syndrome (MERS); SARS-CoV; SARS-CoV-2; and necrotizing enterocolitis (NEC).

[0412] Chronic obstructive pulmonary disease Chronicobstructive pulmonary disease (COPD) is a progressive lung disease, associated with coughing and breathing difficulties. Forms of COPD may include emphysema and chronic bronchitis. Prolonged inflammation in the lungs, typically mediated by eosinophil, neutrophil or macrophage inflammatory cells, can cause remodelling and narrowing of the airways that contribute to reduced lung function. There are currently no cures available for COPD, but the disease can be prevented or managed using suitable treatments. COPD accounted foraround 3.2 million deaths worldwide in 2019.

[0413] Eosinophil extravasation into the tissue of the lungs contributes to the establishment of elevated numbers of local eosinophils in certain population of subjects with COPD. It is known that the number of inflammatory cells present in the lungs correlates well with the severity of disease.

[0414] In the clinical trial results reported in Example 8, the inventors have demonstrated that treatment with SelK2 reduces the number of extravasated eosinophils in lungs of patients with COPD. The same results have also shown that treatment of COPD patients with SelK2 brings about a reduction in number of neutrophils present.

[0415] The skilled person will appreciate that these results, involving a reduction in the numbers of two types of white blood cells that are considered to play an essential role in driving the inflammation underlying COPD, illustrate that SelK2 (and hence pharmaceutical compositions of the invention comprising high concentrations of SelK2, orclosely related antibody fragments or variants thereof) are able to provide effective therapies for the treatment of COPD.

[0416] In view of the above, it will be recognised that the use of pharmaceutical compositions of the invention, or of the methods of treatment or medical uses of the invention, in the prevention or treatment of COPD represent particularly suitable embodiments of these various aspects of the invention.

[0417] A suitable embodiment of the third aspect of the invention, provides a method of preventing or treating COPD in a subject in need thereof, the method comprising providing a therapeutically effective amount of a pharmaceutical composition in accordance with the invention to the subject.

[0418] A suitable embodiment of the fourth aspect of the invention provides a pharmaceutical composition in accordance with the invention for use in the prevention or treatment of COPD. Respiratory diseases or conditions

[0419] Given the successes observed using SelK2 in both asthma and COPD, it will be recognised that the use of pharmaceutical compositions of the invention (comprising high concentrations of SelK2, or closely related antibody fragments or variants thereof) will be well suited to use in the context of prevention or treatment of respiratory diseases or conditions. In particular, pharmaceutical compositions or methods of treatment or medical uses of the invention may be employed in the prevention or treatment of respiratory diseases or conditions associated with inflammation. The pharmaceutical compositions or methods of treatment or medical uses of the invention may be employed in the prevention or treatment of respiratory diseases or conditions associated with the activity of eosinophils or neutrophils.

[0420] By way of example, the pharmaceutical compositions of the invention, or the methods of treatment or medical uses of the invention, may be employed in the prevention or treatment of a respiratory disease or condition selected from the group consisting of: eosinophilic pneumonia (whether chronic or acute); simple pulmonary eosinophilia (Loeffler syndrome); allergic bronchopulmonary aspergillosis; eosinophilic granulomatosis with polyangiitis; and iatrogenic diseases, such as eosinophilia caused by exposure to sulphonamides.

[0421] Sickle cell disease (SCD)

[0422] SCD is a genetic blood disorder characterized by the presence of abnormal haemoglobin known as haemoglobin S (HbS). This abnormal haemoglobin causes red blood cells to take on a characteristic sickle or crescent shape, leading to a range of health complications, including pain crises, anemia, and organ damage.

[0423] Key pathological features of SCD are vaso-occlusion, where sickle-shaped red blood cells clump together and block small blood vessels, and chronic inflammation, associated with an increase in eosinophil counts or activation of eosinophils. These processes lead to symptoms of SCD such as pain crises and tissue damage.

[0424] The formation of complexes between PSGL-1 and P-selectin plays a major role in both of these processes. As noted above, binding of PSGL-1 to its ligands plays an important role in the extravasation and influx of inflammatory cells associated with inflammation. Furthermore, upregulation of P-selectin in endothelial cells and platelets contributes to the cell-to-cell interactions involved in the pathogenesis of vaso-occlusion and pain crisis in SCD. In view of the demonstrated ability of the antibodies incorporated in the pharmaceutical compositions of the invention to inhibit formation of complexes between PSGL-1 and P- selectin and also to disrupt such complexes that have already formed, it will be recognised that the pharmaceutical compositions of the invention are well suited to use in the context of prevention or treatment of SCD or its symptoms. In particular, the pharmaceutical compositions of the invention may be employed in the prevention or treatment of SCD or symptoms of SCD.

[0425] A suitable embodiment of the third aspect of the invention provides a method of preventing or treating SCD in a subject in need thereof, the method comprising providing a therapeutically effective amount of a pharmaceutical composition of the invention to the subject.

[0426] A suitable embodiment of a fourth aspect of the invention provides a pharmaceutical composition in accordance with the invention for use in the prevention or treatment of SCD.

[0427] In the context of the present disclosure, and particularly with reference to the third and fourth aspects of the invention, preventing ortreating SCD should be taken as encompassing treating or preventing any of the processes, such as vaso-occlusion or chronic inflammation, that give rise to the damaging effects of SCD. Suitably, preventing or treating SCD should be taken as encompassing preventing or treating symptoms associated with SCD. In particular, preventing ortreating SCD should be taken as encompassing preventing ortreating pain crisis associated with SCD.

[0428] The ability of Selk2 to inhibit formation of complexes between PSGL-1 and P-selectin is indicative that compositions, methods of treatment and medical uses of the invention would be useful in preventing SCD, or symptoms of SCD such as pain crisis, before the disease or symptoms occur. The ability of SelK2 to disrupt complexes between PSGL-1 and P-selectin that have already formed, is indicative that compositions, methods of treatment and medical uses of the invention would be useful in treating SCD, or symptoms of SCD such as pain crisis, once the disease or symptom is already underway. Given the distress caused to patients by SCD pain crisis, it will be recognised that agents able to treat patients undergoing such crises, and thereby provide relief from the pain, are highly desirable.

[0429] Treatment of diseases or conditions associated with antagonising PSGL-1 signalling

[0430] The skilled person will be aware of a wide range of diseases orconditions that are associated with PSGL-1 signalling. Such diseases orconditions maybe caused orexacerbated by PSGL- 1 activity and / or PSGL-1 intracellular signalling, e.g., enhancing T-cell exhaustion. Examples of such diseases or conditions are considered in more detail over the following pages. The pharmaceutical composition of the invention may be used in the treatment of such diseases or conditions.

[0431] Conditions to be treated

[0432] Activation of PSGL-1 is associated with many biological processes. Accordingly, the ability to antagonise PSGL-1 activity and / or reduce T-cell exhaustion allows modulation of these processes for clinical purposes. As set out herein, the inventors demonstrate the ability of antibodies for use in the pharmaceutical composition of the invention, methods of treatment or the invention, or medical uses of the invention to antagonise PSGL-1 intracellular signalling and / or reduce T-cell exhaustion.

[0433] In view of this surprising clinical effectiveness, and of the common role that the binding of PSGL-1 to its ligands and PSGL-1 signalling plays in the initiation orpropagation of damaging biological activities, particularly those associated with T-cell exhaustion, the skilled person will also appreciate that the pharmaceutical composition of the invention, methods of treatment of the invention, and medical uses of the invention may be expected to have clinical applications beyond those directly demonstrated herein.

[0434] Treatment of cancer

[0435] Cancer is a disease characterised by aberrant or uncontrolled cell division and growth, and / or invasion of adjacent tissues. Cancer is the second leading cause of death globally, accounting for an estimated 9.6 million deaths, or 1 in 6 deaths, in 2018.

[0436] The ability of pharmaceutical compositions of the invention to antagonise PSGL-1 activity and / or reduce T-cell exhaustion indicates that these pharmaceutical compositions may suitably be used in the treatment of cancer.

[0437] Accordingly, the use of pharmaceutical compositions of the invention in the methods of treatment and medical uses of the invention for the treatment of cancer (e.g., by antagonising PSGL-1 activity and / or by reducing T-cell exhaustion) constitutes a particularly suitable embodiment of these various aspects of the invention. Suitably, the cancer may be selected from the group consisting of: melanoma, sarcoma, lymphoma, central nervous system (CNS) cancer, CNS germ cell tumour, lung cancer, leukaemia, multiple myeloma, renal cancer, glioma, medulloblastoma, ovarian cancer, breast cancer, prostate cancer, bladder cancer, pancreatic cancer, gastric cancer, head and neck cancer, mesothelioma, non-melanoma skin cancer, and colorectal cancer; optionally wherein (i) the sarcoma is a fibrosarcoma; (ii) the lymphoma is follicular lymphoma, Hodgkin's lymphoma, or non-Hodgkin's lymphoma; (iii) the lung cancer is non-small cell lung cancer; (iv) the renal cancer is renal cell carcinoma; (v) the ovarian cancer is epithelial ovarian cancer; (vi) the breast cancer is triple negative breast cancer; (vii) the non-melanoma skin canceris merkel cell carcinoma; and / or (viii) the colorectal cancer is colon adenocarcinoma.

[0438] In another embodiment, the cancer is selected from the group consisting of: undifferentiated carcinoma; giant and spindle cell carcinoma; small cell carcinoma; papillary carcinoma; squamous cell carcinoma; lymphoepithelial carcinoma; basal cell carcinoma; pilomatrix carcinoma; transitional cell carcinoma; papillary transitional cell carcinoma; adenocarcinoma; gastrinoma; cholangiocarcinoma; hepatocellular carcinoma; combined hepatocellular carcinoma and cholangiocarcinoma; trabecularadenocarcinoma; adenoid cystic carcinoma; adenocarcinoma in adenomatous polyp; familial polyposis coli adenocarcinoma; solid carcinoma; carcinoid tumour; branchiolo-alveolar adenocarcinoma; papillary adenocarcinoma; chromophobe carcinoma; acidophil carcinoma; oxyphilic adenocarcinoma; basophil carcinoma; clear cell adenocarcinoma; granular cell carcinoma; follicular adenocarcinoma; papillary and follicular adenocarcinoma; non-encapsulating sclerosing carcinoma; adrenal cortical carcinoma; endometroid carcinoma; skin appendage carcinoma; apocrine adenocarcinoma; sebaceous adenocarcinoma; ceruminous; adenocarcinoma; mucoepidermoid carcinoma; cystadenocarcinoma; papillary cystadenocarcinoma; papillary serous cystadenocarcinoma; mucinous cystadenocarcinoma; mucinous adenocarcinoma; signet ring cell carcinoma; infiltrating duct carcinoma; medullary carcinoma; lobular carcinoma; inflammatory carcinoma; mammary Paget's disease; acinar cell carcinoma; adenosquamous carcinoma; thymoma; ovarian stromal tumour; thecoma; granulosa cell tumour; roblastoma; Sertoli cell carcinoma; Leydig cell tumour; lipid cell tumour; paraganglioma; extra-mammary paraganglioma; pheochromocytoma; glomangiosarcoma; amelanotic melanoma; superficial spreading melanoma; epithelioid cell melanoma; blue nevus; fibrous histiocytoma; myxosarcoma; liposarcoma; leiomyosarcoma; rhabdomyosarcoma; embryonal rhabdomyosarcoma; alveolar rhabdomyosarcoma; stromal sarcoma; mullerian mixed tumour; nephroblastoma; hepatoblastoma; carcinosarcoma; mesenchymoma; brenner tumour; phyllodes tumour; synovial sarcoma; dysgerminoma; embryonal carcinoma; teratoma; struma ovarii; choriocarcinoma; mesonephroma; hemangiosarcoma; hemangioendothelioma; kaposi's sarcoma; hemangiopericytoma; lymphangiosarcoma; osteosarcoma; juxtacortical osteosarcoma; chondrosarcoma; chondroblastoma; mesenchymal chondrosarcoma; giant cell tumour of bone; Ewing's sarcoma; odontogenic tumour; ameloblastic odontosarcoma; ameloblastoma; ameloblastic fibrosarcoma; pinealoma; chordoma; ependymoma; astrocytoma; protoplasmic astrocytoma; fibrillary astrocytoma; astroblastoma; glioblastoma; oligodendroglioma; oligodendroblastoma; primitive neuroectodermal; cerebellar sarcoma; ganglioneuroblastoma; neuroblastoma; retinoblastoma; olfactory neurogenic tumour; meningioma; neurofibrosarcoma; neurilemmoma; granular cell tumour; paragranuloma; small lymphocytic lymphoma; diffuse large cell lymphoma; mycosis fungoides; histiocytosis; mast cell sarcoma; immunoproliferative small intestinal disease; lymphoid leukaemia; plasma cell leukaemia; erythroleukemia; lymphosarcoma cell leukaemia; myeloid leukaemia; basophilic leukaemia; eosinophilic leukaemia; monocytic leukaemia; mast cell leukaemia; megakaryoblastic leukaemia; myeloid sarcoma; and hairy cell leukaemia.

[0439] The pharmaceutical compositions of the invention are likely to provide treatment that will be effective against primary tumours. A primary tumour is the original, or first, tumour of a cancer in a subject’s body, defined by a tumour growing at the anatomical site where tumour progression begins and proceeds to yield a cancerous mass. This can be distinguished from metastasis, whereby cancer cells from a primary tumour spread to other organs or other parts of the body to form secondary tumours. Cancers are often named according to the tissue site where the primary tumouris located. Thus, in some embodiments, the cancerto be treated by the pharmaceutical composition of the invention in a method of treatment or medical use of the invention is a primary tumour.

[0440] The pharmaceutical compositions of the invention are likely to provide treatment that will be effective against solid tumours. A solid tumour is an abnormal mass of tissue that usually does not contain cysts or liquid areas and usually do not have a myeloid- or lymphoid-origin. Solid tumours may be benign (not cancerous), or malignant. Examples of solid tumours are sarcomas, carcinomas, and lymphomas. In contrast, leukaemia generally does notform solid tumours. Thus, in some embodiments, the cancer to be treated by the pharmaceutical composition of the invention in a method of treatment or medical use of the invention is a solid tumour.

[0441] The pharmaceutical composition of the invention are likely to provide treatment that will be effective against liquid tumours. Liquid tumours refer to tumours that originates from myeloid or lymphoid cells. These cancers generally affect the bone marrow, the blood cells and the lymphatic system (also called “blood cancers”). Examples of liquid tumours are leukaemia, lymphoma and myeloma. Thus, In some embodiments, the cancer to be treated by the pharmaceutical composition of the invention in a method of treatment or medical use of the invention is a liquid tumour.

[0442] The pharmaceutical compositions of the invention are likely to provide treatment that will be effective in patients with relapsed or refractory cancers. Treatment of this sort, using a pharmaceutical composition of the invention, may bring about a reduction in tumour size and / or a reduction in clinically relevant biomarkers, either of which may be associated with more favourable prognosis. Furthermore, treatment with a pharmaceutical composition of the invention, may help to maintain a reduction in the size of tumours in patients with relapsed or refractory cancer. Accordingly, treatment using a pharmaceutical composition of the invention may achieve a high, durable Disease Control Rate (DCR) in patients with relapsed or refractory cancers. Without wishing to be bound by any hypothesis, the inventors believe that the ability of the pharmaceutical compositions of the invention to sensitize cancers to immunotherapy or immunomodulatory agents and / or reduce T-cell exhaustion, contributes to the therapeutic utility of these agents in the treatment of cancers, and in particular relapsed or refractory cancer. Thus, in some embodiments, the cancer to be treated by the pharmaceutical composition of the invention in a method of treatment or medical uses of the invention is a relapsed or refractory cancer.

[0443] In some embodiments, it may be desirable to sensitize a cancer to treatment with an immunotherapy or immunomodulatory agent. Immunotherapy is the treatment of disease by activating or suppressing the immune system. Immunomodulation relates to treatment of disease by changing or altering the immune system. The skilled person will be familiar with a range of immunotherapy or immunomodulatory agents suitable for the treatment of cancer. In some embodiments, the immunotherapy or immunomodulatory agent may be an antibody. In some embodiments, the immunotherapy or immunomodulatory agent is an immune checkpoint inhibitor. In some embodiments, the immunotherapy agent is an inhibitor of CTLA- 4, PD-1 , PD-L1 , PD-L2. TIM-3, LAGS, TIGIT, CD160, B7H3, B7H4, B7H6, BTLA, VISTA, LAIR1 , and / or LAIR2. Examples of such checkpoint inhibitors include ipilimumab and nivolumab.

[0444] Thus, in some embodiments, provided herein is a method of sensitizing cancer to treatment with an immunotherapy agent, the method comprising providing a therapeutically effective amount of a pharmaceutical composition ofthe invention to a subject in need thereof. In some embodiments, the cancer is a solid tumour. In other embodiments, the cancer is a liquid tumour. In a preferred embodiment, the cancer is a primary tumour. In some embodiments, the cancer comprises dormant cancer cells prior to sensitization, in the context of the invention, “dormant cancer relates to a tumourwhose growth is arrested, lacks proliferative capacity, and / or is in a state of quiescence.

[0445] In some embodiments, it may be desirable to maintain stable disease rather than achieving cancer clearance. Thus, in some embodiments, provided herein is a method of maintaining the size of a tumour, the method comprising providing a therapeutically effective amount of a pharmaceutical composition of the invention to a subject in need thereof.

[0446] In some embodiments, the cancer may already be reducing or shrinking, or have reduced or shrunken (e.g., by way of an anti-cancer agent, such as chemotherapy, radiotherapy or immunotherapy), and it may be desirable to support, maintain or enhance this reduction or shrinkage. Thus, in some embodiments, provided herein is a method of maintaining a reduction in size of a tumour, the method comprising providing a therapeutically effective amount of a pharmaceutical composition of the invention to a subject in need thereof.

[0447] In some embodiments, the pharmaceutical compositions of the invention are provided in combination with a chemotherapy agent. Suitable chemotherapy agents are well known in the art and to the skilled person. For instance, chemotherapy agents suitable for use in accordance with this aspect of the invention include altretamine, bendamustine, busulfan, carboplatin, chlorambucil, cisplatin, cyclophosphamide, dacarbazine, ifosfamide, mechlorethamine, melphalan, oxaliplatin, procarbazine, temozolomide, thiotepa, trabectedin, carmustine, lomustine, streptozocin, 5-fluorouracil, 6-mercaptopurine, azacitidine, capecitabine, cladribine, clofarabine, cytarabine, decitabine, floxuridine, fludarabine, gemcitabine, hydroxyurea, methotrexate, nelarabine, pemetrexed, pentostatin, pralatrexate, thioguanine, trifluridine / tipiracil combination, etoposide, irinotecan, irinotecan liposomal, mitoxantrone, teniposide, topotecan, cabazitaxel, docetaxel, nab-paclitaxel, paclitaxel, vinblastine, vincristine, vincristine liposomal, vinorelbine, daunorubicin, doxorubicin, doxorubicin liposomal, epirubicin, idarubicin, mitoxantrone, valrubicin, bleomycin, dactinomycin, mitomycin-c, all-trans-retinoic acid, arsenic trioxide, asparaginase, eribulin, ixabepilone, mitotane, omacetaxine, pegaspargase, procarbazine, romidepsin, and vorinostat.

[0448] Treatment of infection, e.g., chronic infection

[0449] Infection is characterised by the invasion and multiplication of an infectious agent (pathogen) in a subject. Infectious diseases rank among the leading causes of death globally. The ability of the pharmaceutical compositions of the invention to antagonise PSGL-1 intracellular signalling indicates that these agents may suitably be used in the treatment of infection, e.g., chronic infection. In particular, the ability of the pharmaceutical compositions of the invention to reduce T-cell exhaustion indicates that these agents may suitable be used in the treatment of infection, and specifically, chronic infection.

[0450] Suitably, the infection may be viral, bacterial, fungal, parasitic, or protozoan. The infection may be acute or chronic. Acute infection is characterized by sudden or rapid onset of disease due to infection. Acute infections are well-known to the skilled person, and include infection with influenza and SARS-CoV-2. Chronic infection, or persistent infection, is characterized by the continued presence of infectious agentfollowingthe primary infection and may include chronic or recurrent disease. Chronic infection may be symptomatic or latent and are often resistant to first-line medical treatments. Chronic infections may be able to be eradicated after treatment, or may be a life-long condition.

[0451] Accordingly, the use of pharmaceutical compositions of the invention in the methods of treatment and medical uses of the invention for the treatment of infection, e.g., chronic infection (e.g., by antagonising PSGL-1 activity and / or by reducing T-cell exhaustion) constitutes a particularly suitable embodiment of these various aspects of the invention.

[0452] Suitably, the chronic infection may comprise infection with a viral agent selected from the group of: HIV, Hepatitis B, Hepatitis C, herpes simplex virus, varicella zoster virus, cytomegalovirus, HTLV1 , mumps virus, rubella virus, measles virus, poliovirus, SARS-CoV-2 (e.g., “long COVID”), and Epstein-Barr virus. Alternatively, the chronic infection may comprise infection with a bacterial agent selected from the group of: Mycobacterium tuberculosis, Helicobacter pylori, Salmonella Typhi, Treponema pallidum, Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus (e.g., MRSA), Hemophilus influenza, and Mycobacterium leprae. Alternatively, the chronic infection may comprise infection with a fungal species selected from the group of: Candida (e.g., Candidaalbicans), Malassezia, Sporothrix, Histoplasma, Coccidioides, Blastomyces, Aspergillus, Pneumocystis (e.g., Pneumocystis jirovecii), Rhizopus, Mucor, and Cryptococcus (e.g., Cryptococcus neoformans and Cryptococcus gattii). Alternatively, the chronic infection may comprise infection with a parasite (e.g., Helminth) or protozoa selected from the group of: Plasmodium (e.g., Plasmodium falciparum, Plasmodium malariae, Plasmodium knowlesi, Plasmodium vivax, and Plasmodium ovale), Giardia (e.g., Giardia lamblia), Toxoplasma (e.g., Toxoplasma gondii), Cyclospora, Entamoeba, Isospora, Blastocystis, Strongyloides, Schistosoma (e.g., Schistosoma haematobium, Schistosoma mansoni, and Schistosoma japonicum), Leishmania, Trypanosoma, Opisthorchis viverrini, Clonorchis sinensis, Heligmosomoides polygyrus, Taenia (e.g., Taenia crassicep and Taenia solium), Ascaris lumbricoides, Nercator americanus, Trichomonas vaginalis, Entamoeba histolytica, Wuchereria bancrofti, Onchocerca volvulus, Enterobius vermicularis, and Trichuris trichiura.

[0453] In some embodiments, chronic infection is characterised by infectionwith an infectious agent which is well-known to the skilled person for its capacity for persistence or latency, or difficulty to eradicate following infection, e.g., DNA viruses (e.g., HSV), retroviruses (e.g., HIV), tuberculosis, and malaria. In otherembodiments, chronic infection is characterised by infection with an infectious agent for at least one, two, three, four, five, six, seven, eight, nine, ten, eleven or twelve months. In otherembodiments, chronic infection is characterised by infection with an infectious agent for at least two, three, four, five, six, seven, eight, nine, ten, or more years.

[0454] In some embodiments, a pharmaceutical composition of the invention is provided in combination with an anti-infective agent. Anti-infective agents encompass anti-viral, anti- bacterial, anti-fungal, anti-parasitic and anti-protozoan agents. Suitable anti-infective agents are well known in the art and to the skilled person. An example of an anti-infective agent is an antibiotic.

[0455] Combination therapy with immune checkpoint inhibitors

[0456] As discussed elsewhere in the disclosure, reversal of T-cell exhaustion can be linked to disease resolution against infection, e.g. chronic infection, and cancer, and this is an objective of immunotherapies which utilise checkpoint inhibitor therapy. PSGL-1 has been identified as a potent negative regulator of T-cell function in infection, e.g. chronic infection, and cancer. Accordingly, antibodies targeting PSGL-1 may reduce T-cell exhaustion.

[0457] The parental antibody from which SelK2 is derived has been found to be effective at slowing melanoma growth in a highly aggressive B16 mouse melanoma model which is resistant to anti-PD-1 and anti-PD-L1. This effect was characterised by increased T-cell activation, proliferation, and effector functions. Similarly, growth of the anti-PD1 resistant D4M-3A melanoma tumour in a mouse model was also delayed following administration of the parental antibody. Of note, the combination of the parental antibody with anti-PD-1 therapy increased the median overall survival of the treated mice. The present inventors have demonstrated that Selk2, has higher affinity for PSGL-1 than their parental antibody. Thus, it is expected that pharmaceutical compositions of the invention (comprising Selk2 or closely related antibody fragments or variants) will retain the ability to inhibit melanoma growth in PD-1 resistant tumours and increase effectiveness in combination with anti-PD-1 therapy.

[0458] The findings suggest that using pharmaceutical compositions of the invention to target PSGL- 1 , either alone or in combination with standard therapies, may represent a new therapeutic approach to control tumours.

[0459] Accordingly, in one embodiment, provided herein is a pharmaceutical composition of the invention for use as a medicament for the treatment of cancer (e.g., by antagonising PSGL-1 activity and / or reducing T-cell exhaustion), wherein the subject has a cancer resistant to an immune checkpoint inhibitor. In another embodiment, provided herein is a pharmaceutical composition of the invention for use as a medicament for the treatment of infection, e.g. chronic infection, (e.g., by antagonising PSGL-1 activity and / or reducing T-cell exhaustion), wherein the subject has an infection, e.g. chronic infection, resistant to an immune checkpoint inhibitor.

[0460] In the context of the invention, “resistance” or“resistant” to a therapy, e.g., immune checkpoint inhibitor therapy, relates to a cancer or infection, e.g. chronic infection, that does not respond to, or is no longer responding to, immune checkpoint inhibitor treatment. This may include recurrence, relapse and / or refractory cancer.

[0461] In some embodiments, the pharmaceutical compositions of the invention are suitable for administration to a subject with a cancer, wherein the subject has previously been administered an immunotherapy. In anotherembodiment, the pharmaceutical compositions of the invention are suitable for administration to a subject with an infection, e.g., chronic infection, wherein the subject has previously been administered an immunotherapy.

[0462] In an embodiment, provided herein is a pharmaceutical composition of the invention for use as a medicament for the treatment of cancer by antagonising PSGL-1 activity, in combination with an immunotherapy. In another embodiment, provided herein is a pharmaceutical composition of the invention for use as a medicament for the treatment of infection, e.g. chronic infection by antagonising PSGL-1 activity, in combination with an immunotherapy. In some embodiments, the immunotherapy agent may be an antibody. In a preferred embodiment, the immunotherapy is an immune checkpoint inhibitor.

[0463] In some embodiments, the immune checkpoint inhibitoris an inhibitor of CTLA-4, PD-1 , PD- L1 , PD-L2, TIM-3, LAG3, TIGIT, CD160, B7H3, B7H4, B7H6, BTLA, VISTA, LAIR1 , and / or LAIR2. In some embodiments, the immune checkpoint inhibitor is an inhibitor of CTLA-4, PD- 1 , and / or PD-L1.

[0464] In the context of the invention, the meaning of a “combination" of the pharmaceutical composition of the invention for use in the methods of treatment or medical uses of the invention with another therapy, e.g., an immunotherapy, will be known to the skilled person in the art. A combination therapy may refer to the presence of both agents (e.g., and antibody, fragmentorvariantofa pharmaceutical composition ofthe invention, and afurthertherapeutic agent) in therapeutically effective amounts in the subject's circulation at the same time point. Combination therapy may relate to simultaneous or sequential administration.

[0465] In some embodiments, the pharmaceutical composition of the invention for use in the methods of treatment and medical uses of the invention is administered simultaneously with the immunotherapy. In another embodiment, the pharmaceutical composition of the invention for use in the methods of treatment and medical uses of the invention is administered sequentially with the immunotherapy. In a suitable embodiment, the pharmaceutical composition of the invention for use in the methods of treatmentand medical uses of the invention is administered separately to the immunotherapy. Such separate administration may be within a sufficient time-period of each other such that both agents are present (e.g., in a therapeutically effective amount) in the subject’s circulation at the same time point.

[0466] In the context of the invention, “simultaneous” administration encompasses administration of one agent within one hour of the second agent.

[0467] In some embodiments, the pharmaceutical composition of the invention for use in the methods of treatment and medical uses of the invention is administered one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, twenty, twenty-one, twenty-two, twenty-three, twenty-four, or more hours prior to the administration of the immunotherapy. In some embodiments, the pharmaceutical composition of the invention for use in the methods of treatment and medical uses of the invention is administered one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, twenty, twenty-one, twenty-two, twenty-three, twenty-four, twenty-five, twenty-six, twenty-seven, twenty-eight, twenty-nine, thirty, or more days prior to the administration of the immunotherapy, in some embodiments, the pharmaceutical composition of the invention for use in the methods of treatment and medical uses of the invention is administered one, two, three, four, or moreweeks priorto the administration of the immunotherapy.

[0468] In alternative embodiments, the pharmaceutical composition of the invention for use in the methods of treatment and medical uses of the invention is administered one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, twenty, twenty-one, twenty-two, twenty-three, twenty-four, or more hours following the administration of the immunotherapy. In some embodiments, the pharmaceutical composition of the invention for use in the methods of treatment and medical uses of the invention is administered one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, twenty, twenty-one, twenty- two, twenty-three, twenty-four, twenty-five, twenty-six, twenty-seven, twenty-eight, twenty- nine, thirty, or more days following administration of the immunotherapy, in some embodiments, the pharmaceutical composition of the invention for use in the methods of treatment and medical uses of the invention is administered one, two, three, four, or more weeks following administration of the immunotherapy.

[0469] In a preferred embodiment, the use of the combination results in enhanced therapeutic efficacy relative to the use of the immune checkpoint inhibitor alone. For instance, the therapeutic efficacy is increased, relative to the use of an immune checkpoint inhibitoralone, by at least 10%, at least 11 %, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17%, at least 18%, at least 19%, at least 20%, at least 21 %, at least 22%, at least 23%, at least 24%, at least 25%, at least 26%, at least 27%, at least 28%, at least 29%, at least 30%, at least 31 %, at least 32%, at least 33%, at least 34%, at least 35%, at least 36%, at least 37%, at least 38%, at least 39%, at least 40%, at least 41 %, at least 42%, at least 43%, at least 44%, at least 45%, at least 46%, at least 47%, at least 48%, at least 49%, at least 50%, at least 51 %, at least 52%, at least 53%, at least 54%, at least 55%, at least 56%, at least 57%, at least 58%, at least 59%, at least 60%, at least 61 %, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71 %, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or even by 100%. Therapeutically effective amounts of compositions of the invention

[0470] The methods of treatment of the invention involve the provision of therapeutically effective amounts of compositions of the invention. Such therapeutically effective amounts of the requisite compositions may be provided in a single incidence of treatment, or may be accumulated over the course of a number of incidences of treatment.

[0471] For the purposes of the present disclosure, a therapeutically effective amount of a pharmaceutical composition of the invention may be taken as referring to a quantity or dose of the pharmaceutical composition that is sufficient to produce a desired therapeutic or beneficial effect in a patient or individual. Atherapeutically effective amount may be an amount sufficient to reduce or delay the onset of a disease or condition, or an amount sufficient to reduce or delay the onset of one or more symptoms of a disease or condition. Atherapeutically effective amount may be an amount sufficient to provide at least partial relief in respect of a disease or condition, or an amount sufficient to provide at least partial relief in respect of one or more symptoms of a disease or condition. Atherapeutically effective amount may be an amount sufficientto entirely preventthe onset of a disease orcondition, oran amount sufficient to entirely prevent the onset of one or more symptoms of a disease or condition. A therapeutically effective amount may be an amount sufficient to entirely alleviate a disease or condition, or an amount sufficient to entirely alleviate one or more symptoms of a disease or condition. Other specific considerations regarding therapeutically effective amounts of pharmaceutical compositions will be recognised by those skilled in the art, as considered further below.

[0472] It is an advantage of the pharmaceutical compositions of the invention that the high concentrations of antibodies (or fragments or variants thereof) that may be incorporated in them makes it possible to provide a therapeutically effective amount of the antibody to a subject in a relatively small volume of the composition. This makes it simple to provide a therapeutically effective amount in a single incidence of administration.

[0473] Conventionally, a therapeutically effective amount may be presented as a dose per unit weight of the subject being treated. With this knowledge, and details of the concentration of an antibody, fragment or variant in the pharmaceutical composition in question, the volume of the pharmaceutical composition that must be administered in order to provide a therapeutically effective amount may readily be calculated. Typically, required dosages may be determined by a clinician with responsibility for the subject’s care. A therapeutically effective amount of a pharmaceutical composition of the invention required by a subject may be selected based upon a number of factors. These include, but are not limited to: the weight of the subject; the age of the subject; the nature of the condition or disease to be treated; the severity of the condition or disease to be treated; whetherthe pharmaceutical composition is being used for prevention ortreatment; the stability of the antibody incorporated in the pharmaceutical composition; the number of incidences of treatment desired; and the period between incidences of administration.

[0474] In a suitable embodiment, a therapeutically effective amount of an antibody, fragment or variant, to be provided by a pharmaceutical composition of the invention in an incidence of treatment may be between about 1 mg / kg and about 50 mg / kg.

[0475] The Examples demonstrate the effectiveness of treatment regimens in which an antibody of the invention is administered at a dose of 7.5 mg / kg body weight (for example with a three week period between incidences of administration). Such regimens may be of benefit in the treatment of patients with respiratory disorders, such as asthma or COPD.

[0476] For example, an antibody may be administered to a patient in a therapeutically effective amount of approximately 1 mg / kg, approximately 1.5 mg / kg, approximately 2 mg / kg, approximately 2.5 mg / kg, approximately 3 mg / kg, approximately 3.5 mg / kg, approximately 4 mg / kg, approximately 4.5 mg / kg, approximately 5 mg / kg, approximately 5.5 mg / kg, approximately 6 mg / kg, approximately 6.5 mg / kg, approximately 7 mg / kg, approximately 7.5 mg / kg, approximately 8 mg / kg, approximately 8.5 mg / kg, approximately 9 mg / kg, approximately 9.5 mg / kg, approximately 10 mg / kg, approximately 10.5 mg / kg, approximately 11 mg / kg, approximately 12 mg / kg, approximately 13 mg / kg, approximately 14 mg / kg, approximately 15 mg / kg, approximately 20 mg / kg, approximately 25 mg / kg, or approximately 30 mg / kg.

[0477] In a suitable embodiment, a therapeutically effective amount of an antibody is between approximately 10 mg and approximately 5000 mg. For example, in a suitable embodiment, a therapeutically effective amount of an antibody is between approximately 100 mg and approximately 1500 mg, for example between approximately 200 mg and approximately 1000 mg, or between approximately 450 mg and approximately 750 mg. Suitably, a therapeutically effective amount of an antibody of the invention is approximately 600 mg. With knowledge of the concentration of the antibody, fragment or variant in the composition of the invention, it will be a simple matter for the skilled person to determine the necessary quantities of a composition to be administered in order to achieve any of the therapeutically effective amounts described in this section of the specification.

[0478] Routes of administration

[0479] When practicing the methods of treatment or medical uses of the invention, pharmaceutical compositions of the invention may be provided to a subject by any suitable route of administration. In particular, the pharmaceutical compositions of the invention may be administered by means of injection for example by means of subcutaneous injection, intravenous injection or intramuscular injection.

[0480] Suitably, a pharmaceutical composition of the invention is provided to the subject by subcutaneous (i.e. beneath the skin) administration. Forsuch purposes, the a pharmaceutical composition of the invention may be injected using a syringe and needle. However, other devices for administration of the formulation are known to those skilled in the art. Suitably a pharmaceutical composition of the invention may be provided to a recipient by means of a device selected from the group consisting of: an injector pen, an auto-injector device, an injection port, a needle-free injector, and a subcutaneous patch delivery system. Indeed, in a suitable embodiment, a pharmaceutical composition of the invention may be provided in a pre- loaded injection device selected from the group consisting of: a syringe, an injector pen, an auto-injector device, an injection port, a needle-free injector, and a subcutaneous patch delivery system.

[0481] In a suitable embodiment, a pharmaceutical composition of the invention is provided to the subject by subcutaneous injection. The suitability of the pharmaceutical compositions for such administration has been demonstrated in syringeability tests in a laboratory setting, as described further in the Examples.

[0482] STRUCTURE OF SELK2

[0483] As referred to above, SelK2 is a humanised anti-PSGL-1 lgG2 antibody that is an exemplar of the antibodies that may be incorporated in pharmaceutical compositions of the invention. Its sequence also serves as a reference antibody for antibody fragments and variants that may be incorporated in such pharmaceutical compositions. As set out elsewhere, SelK2 consists of the antibody heavy chain set out in SEQ ID NO: 1 and the antibody light chain set outin SEQ ID NO: 2. Fragmentsorvariantsofantibodies that may be incorporated in the pharmaceutical compositions of the invention share at least 90% sequence identity with the SelK2 antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2.

[0484] Within this broad definition, there are a number of further ways in which antibody fragments and variants that may be incorporated in the pharmaceutical composition of the invention can be defined with reference to the amino acids sequence of SelK2. Useful examples of such definitions are set out below.

[0485] Variable regions of antibodies (such as SelK2), or fragments or variants thereof

[0486] An antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may comprise one or both of the variable regions of the SelK2 antibody. Since the variable regions of the antibody confer its specificity, such antibody fragments or variants may be expected to share the specificity of SelK2.

[0487] SEQ ID NO: 3 is the heavy chain variable region of the heavy chain sequence set out in SEQ ID NO: 1. In a suitable embodiment, an antibody, fragment, orvariantthat may be incorporated in a pharmaceutical composition of the invention comprises a heavy chain variable region set out in SEQ ID NO: 3.

[0488] SEQ ID NO: 4 is the light chain variable region of the light chain sequence set out in SEQ ID NO: 2. In a suitable embodiment, an antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention comprises a light chain variable region set out in SEQ ID NO: 4.

[0489] In a suitable embodiment, an antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may comprise both the heavy chain variable region set out in SEQ ID NO: 3 and the light chain variable region set out in SEQ ID NO: 4.

[0490] In the case of antibody fragments comprising SEQ ID NO: 3 and / or SEQ ID NO: 4, it will be appreciated that the portions truncated as compared to the reference antibody will be from areas other than the variable region. Similarly, in the case of variants comprising SEQ ID NO: 3 and / or SEQ ID NO: 4, it will be appreciated that the sequence alterations as compared to the reference antibody will be made areas other than the variable region. Alternatively, an antibody fragment or variant suitable for incorporation in a pharmaceutical composition of the invention may comprise a heavy or light chain variable region that shares less than 100% identity with those set out in SEQ ID NO: 3 and SEQ ID NO: 4.

[0491] For example, an antibody fragment or variant suitable for incorporation in a pharmaceutical composition of the invention may comprise a heavy or light chain variable region that shares at least 99% identity with the heavy chain variable region set out in SEQ ID NO: 3 and / or with the light chain variable region set out in SEQ ID NO: 4, or that shares at least 98%, at least 97%, at least 96%, at least 95%, at least 94%, at least 93%, at least 92%, at least 91 %, or at least 90% identity with the heavy chain variable region set out in SEQ ID NO: 3 and / or with the light chain variable region set out in SEQ ID NO: 4.

[0492] If a fragment or variant shares a high degree of identity with SelK2 outside the variable regions, it may share lower identity within the variable regions. For example, a suitable antibody fragment or variant sharing a high degree of identity with SelK2 outside the variable regions may share up to 70% identity with SEQ ID NO: 3 and / or SEQ ID NO: 4, up to 75% identity with SEQ ID NO: 3 and / or SEQ ID NO: 4, up to 80% identity with SEQ ID NO: 3 and / or SEQ ID NO: 4, up to 85% identity with SEQ ID NO: 3 and / or SEQ ID NO: 4, or up to 90% identity with SEQ ID NO: 3 and / or SEQ ID NO: 4.

[0493] Constant regions of antibodies (such as SelK2), or fragments or variants thereof

[0494] An antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may comprise any desired constant region. A skilled person will readily be able to select a suitable constant region taking into account the requirements of the applications in which the antibody is to be used.

[0495] An antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may comprise the heavy chain and / or light chain constant regions of the SelK2 antibody.

[0496] Merely by way of example, a suitable an antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may comprise a human lgG2 heavy chain constant region. A suitable antibody may comprise a human lgG2 heavy chain constant region of SEQ ID NO: 5. SEQ ID NO: 5 is the heavy chain constant region of the heavy chain sequence set out in SEQ ID NO: 1 . The incorporation of a human lgG2 heavy chain constant region in an antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may be considered favourable for a number of reasons. For example, since the constant region of lgG2 exhibits low binding to Fc receptor, and low binding to complement component 1 q (C1 q), antibodies that incorporate this constant region have a reduced likelihood of inducing antibody effector functions. The human lgG2 heavy chain constant region of SEQ ID NO: 5 has particularly reduced potential for inducing such functions, since it incorporates a modification (the alanine residue at position 201 of SEQ ID NO: 5, replacing the lysine residue found at this position in naturally occurring human lgG2) that further reduces binding to C1 q.

[0497] Suitably an antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention comprises a human kappa light chain constant region. For example, such an antibody may comprise a human kappa light chain constant region of SEQ ID NO: 6. SEQ ID NO: 6 is the light chain constant region of the heavy chain sequence set out in SEQ ID NO: 1 . In a suitable embodiment, a SelK2 antibody of invention comprises a light chain constant region set out in SEQ ID NO: 6.

[0498] In a suitable embodiment, an antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may comprise both the heavy chain constant region set out in SEQ ID NO: 5 and the light chain constant region set out in SEQ ID NO: 6.

[0499] Alternatively, a suitable antibody fragment or variant for incorporation in a pharmaceutical composition of the invention may share at least 99% identity with the heavy chain constant region set out in SEQ ID NO: 5 and / or with the light chain constant region set out in SEQ I D NO: 6, or that shares at least 98%, at least 97%, at least 96%, at least 95%, at least 94%, at least 93%, at least 92%, at least 91 %, or at least 90% identity with the heavy chain constant region set out in SEQ ID NO: 5 and / or with the light chain constant region set out in SEQ I D NO: 6.

[0500] If a fragment or variant shares a high degree of identity with SelK2 outside the constant regions, it may share lower identity within the constant regions. For example, a suitable antibody fragment or variant sharing a high degree of identity with SelK2 outside the constant regions may share up to 70% identity with SEQ ID NO: 5 and / or SEQ ID NO: 6, up to 75% identity with SEQ ID NO: 5 and / or SEQ ID NO: 6, up to 80% identity with SEQ ID NO: 5 and / or SEQ ID NO: 6, up to 85% identity with SEQ ID NO: 5 and / or SEQ ID NO: 6, or up to 90% identity with SEQ ID NO: 5 and / or SEQ ID NO: 6. A suitable antibody fragment or variant may retain the alanine residue at position 201 or SEQ ID NO: 5.

[0501] CDRs of antibodies (such as SelK2), or fragments or variants thereof

[0502] Antibodies, fragments or variants that may be incorporated in a pharmaceutical composition of the invention may be defined with reference to the CDRs that they include. In particular, an antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may be defined with reference to the inclusion of the three CDRs from the reference antibody heavy chain sequence set out in SEQ ID NO: 1 , and the three CDRs from the reference antibody light chain sequence set out in SEQ ID NO: 2. Such antibody fragments or variants will be expected to constitute anti-PSGL-1 antibodies, and so to have utility in one or more of the therapeutic applications described with reference to SelK2.

[0503] Suitably an antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may comprise each of a CDR H1 present in SEQ ID NO: 1 , a CDR H2 present in SEQ ID NO: 1 , and a CDR H3 present in SEQ ID NO: 1 .

[0504] Suitably an antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may comprise each of a CDR L1 present in SEQ ID NO: 2, a CDR L2 present in SEQ ID NO: 2, and a CDR L3 present in SEQ ID NO: 2.

[0505] Suitably an antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may comprise each of a CDR H1 present in SEQ ID NO: 1 , a CDR H2 present in SEQ ID NO: 1 , a CDR H3 present in SEQ ID NO: 1 , a CDR L1 present in SEQ ID NO: 2, a CDR L2 present in SEQ ID NO: 2, and a CDR L3 present in SEQ ID NO: 2.

[0506] Suitably an antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may comprise six CDRs that cumulatively share at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with the sequences of CDR H1 present in SEQ ID NO: 1 , CDR H2 present in SEQ ID NO: 1 , CDR H3 present in SEQ ID NO: 1 , CDR L1 present in SEQ ID NO: 2, CDR L2 present in SEQ ID NO: 2, and CDR L3 present in SEQ ID NO: 2.

[0507] Alternatively, antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may comprise one, two, three, four, five or six CDRs, each of which shares at least at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with the sequences of the corresponding CDRs set out in SEQ ID NO: 1 and SEQ ID NO: 2.

[0508] It will be appreciated that there are a number of different schemes and conventions by which the amino acid residues making up CDRs within an antibody heavy chain or an antibody light chain may be identified. Merely by way of example, these include the Kabat numbering scheme, the Chothia numbering scheme, and the IMGT numbering scheme.

[0509] By way of guidance to the skilled practitioner wishing to practice the present invention, Table 69 lists the CDRs of the exemplary antibody SelK2, and set out in SEQ ID NOs: 1 and 2, as identified by:

[0510] • a combination of the Kabat and Chothia numbering schemes;

[0511] • the Kabat numbering scheme;

[0512] • the IMGT numbering scheme; and

[0513] • the Chothia numbering scheme.

[0514] The CDRs that may be included in an antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may suitably be selected from the sequences of SelK2 set out in Table 69.

[0515] Accordingly, a CDR H1 of an antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may comprise or consist of a sequence selected from the group consisting of: SEQ ID NO: 7; SEQ ID NO: 8; SEQ ID NO: 9; and SEQ ID NO: 10, or that shares at least at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with these sequences. A CDR H2 of an antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may comprise orconsist of asequence selected from the group consisting of: SEQ ID NO: 1 1 ; SEQ ID NO: 12; SEQ ID NO: 13; and SEQ ID NO: 14, or that shares at least at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with these sequences. A CDR H3 of an antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may comprise or consist of a sequence selected from the group consisting of: SEQ ID NO: 15; SEQ ID NO: 16; SEQ ID NO: 17; and SEQ ID NO: 18, or that shares at least at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with these sequences.

[0516] A CDR L1 of an antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention , may comprise or consist of a sequence selected from the group consisting of: SEQ ID NO: 19; SEQ ID NO: 20; SEQ ID NO: 21 ; and SEQ ID NO: 22, or that shares at least at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with these sequences. A CDR L2 of an antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may comprise or consist of a sequence selected from the group consisting of: SEQ ID NO: 23; SEQ ID NO: 24; SEQ ID NO: 25; and SEQ ID NO: 26, or that shares at least at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with these sequences. ACDR L3 of an antibody, fragment, orvariantthatmay be incorporated in a pharmaceutical composition of the invention may comprise or consist of a sequence selected from the group consisting of: SEQ ID NO: 27; SEQ ID NO: 28; SEQ ID NO: 29; and SEQ ID NO: 30, or that shares at least at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with these sequences.

[0517] Suitably, the CDRs included in an antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may each be selected with reference to the same numbering scheme. Alternatively, each of the CDRs of an antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may be independently selected from those set out in T able 69, such that the antibody, fragment, or variant contains a CDR H1 , a CDR H2, a CDR H3, a CDR L1 , a CDR L2, and a CDR L3.

[0518] SEQ ID NO: 7 is the Kabat / Chothia-defined heavy chain CDR H1 of the heavy chain sequence set out in SEQ ID NO: 1 . Suitably, an antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may comprise a Kabat / Chothia-defined heavy chain CDR H1 of SEQ ID NO: 7, or that shares at least 90%, or at least 95%, identity with such a sequence.

[0519] SEQ ID NO: 11 is the Kabat / Chothia-defined heavy chain CDR H2 of the heavy chain sequence set out in SEQ ID NO: 1 . Suitably, an antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may comprises a Kabat / Chothia-defined heavy chain CDR H2 of SEQ ID NO: 1 1 , or that shares at least 90%, or at least 95%, identity with such a sequence.

[0520] SEQ ID NO: 15 is the Kabat / Chothia-defined heavy chain CDR H3 of the heavy chain sequence set out in SEQ ID NO: 1 . Suitably an antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may comprise a Kabat / Chothia- defined heavy chain CDR H3 of SEQ ID NO: 15, or that shares at least 90%, or at least 95%, identity with such a sequence.

[0521] Suitably an antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may comprise each of a Kabat / Chothia-defined CDR H1 of SEQ ID NO: 7, a Kabat / Chothia-defined CDR H2 of SEQ ID NO: 1 1 , and a Kabat / Chothia-defined CDR H3 of SEQ ID NO: 15, or sequences that share at least 90%, or at least 95%, identity with these CDRs.

[0522] SEQ ID NO: 19 is the Kabat / Chothia-defined light chain CDR L1 of the light chain sequence set out in SEQ ID NO: 2. Suitably, an antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may comprise a Kabat / Chothia-defined light chain CDR L1 of SEQ ID NO: 19, or that shares at least 90%, or at least 95%, identity with such a sequence.

[0523] SEQ ID NO: 23 is the Kabat / Chothia-defined light chain CDR L2 of the light chain sequence set out in SEQ ID NO: 2. Suitably, an antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may comprise a Kabat / Chothia-defined light chain CDR L2 of SEQ ID NO: 23, or that shares at least 90%, or at least 95%, identity with such a sequence.

[0524] SEQ ID NO: 27 is the Kabat / Chothia-defined light chain CDR L3 of the light chain sequence set out in SEQ ID NO: 2. Suitably, an antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may comprise a Kabat / Chothia-defined light chain CDR L3 of SEQ ID NO: 27, or that shares at least 90%, or at least 95%, identity with such a sequence.

[0525] Suitably an antibody of the invention, or an antibody for use in the methods of treatment and medical uses of the invention, comprises each of a Kabat-def ined CDR L1 of SEQ ID NO: 19, a Kabat-def ined CDR L2 of SEQ ID NO: 23, and a Kabat-def ined CDR L3 of SEQ ID NO: 27. Suitably an antibody of the invention, or an antibody for use in the methods of treatment and medical uses of the invention, comprises each of a Kabat-defined CDR H1 of SEQ ID NO: 7, a Kabat-defined CDR H2 of SEQ ID NO: 11 , a Kabat-defined CDR H3 of SEQ ID NO: 15, a Kabat-defined CDR L1 of SEQ ID NO: 19, a Kabat-defined CDR L2 of SEQ ID NO: 23, and a Kabat-defined CDR L3 of SEQ ID NO: 27.

[0526] In a suitable embodiment of an antibody fragment or variant that may be incorporated in a pharmaceutical composition of the invention, the CDRs of the antibody may be identical to those of the anti-PSGL-1 antibody defined by SEQ ID NO: 1 and SEQ ID NO: 2, and so identical to the CDRs present in the variable regions of SEQ ID NO: 3 and SEQ ID NO: 4. The CDRs of these sequences are identified in Table 69. In such cases the non-identical residues or regions of the antibody fragment or variant may be confined to the non-CDR portions of the relevant variable regions.

[0527] Thus, an antibody suitable for incorporation in a pharmaceutical composition of the invention may suitably comprise the CDRs of the antibody heavy chain sequence set out in SEQ I D NO: 1 and the CDRs of the antibody light chain sequence set out in SEQ ID NO: 2, and a heavy chain variable region that shares at least 90% identity with the heavy chain sequence set out in SEQ ID NO: 3 and / or a light chain variable region that shares at least 90% identity with the light chain sequence set out in SEQ ID NO: 4. In a suitable embodiment, such an antibody comprises the CDRs of SEQ ID NO: 1 and SEQ ID NO: 2, and a heavy chain variable region that shares at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence of the heavy chain variable region set outin SEQ ID NO: 3 and / ora light chain variable region that shares at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the sequence of the light chain variable region set out in SEQ ID NO: 4.

[0528] Humanised antibodies

[0529] SelK2 is a humanised anti-PSGL-1 antibody. An antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may also be derived from a humanised anti-PSGL-1 antibody, such as SelK2. In a suitable embodiment, an antibody suitable for incorporation in an pharmaceutical composition of the invention may comprise the heavy chain of SEQ ID NO: 1 , the light chain of SEQ ID NO: 2 and human germline acceptor sequences.

[0530] In a suitable embodiment, an antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may comprise the heavy chain of SEQ ID NO: 1 in the human germline acceptor sequence VH3 1 -3 3-30.5 (SEQ ID NO: 23). Suitably the framework 4 region of the heavy chain of such an antibody may be that of human germline heavy joining sequence JH4a.

[0531] In a suitable embodiment, an antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may comprise the light chain of SEQ ID NO: 2 in the human germline acceptor sequence IGKV2-40*01 (SEQ ID NO: 24). Suitably the framework 4 region of the light chain of such an antibody may be that of human germline kappa joining sequence JK5.

[0532] Framework region of SelK2 antibodies of the invention

[0533] An antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may comprise a human framework acceptor sequence. In a suitable embodiment 1 , 2, 3 or 4 framework sequences of the heavy chain variable region may be a human framework sequence. In a suitable embodiment 1 , 2, 3 or 4 framework sequences of the light chain variable region may be a human framework sequence. Such sequences may share at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, identity with the corresponding sequences set out in SEQ ID NO: 1 and SEQ ID NO: 2.

[0534] An antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may comprise a substantially human framework acceptor that incorporates one or more back mutations. Suitably an antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may comprise a substantially human framework acceptor that incorporates a single back mutation. An example of such a framework region is found in SEQ ID NO: 1 . Here the serine residue at position 117 of SEQ ID NO: 1 constitutes a back mutation, replacing the lysine residue found in the naturally occurring human sequence. SEQ ID N0:31 is the framework region FR H1 of the heavy chain sequence set out in SEQ ID NO: 1. In a suitable embodiment, an antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention comprises a heavy chain framework region FR H1 of SEQ ID NO: 31 or that shares at least 90%, or at least 95%, identity with such a sequence.

[0535] SEQ ID NO:32 is the framework region FR H2 of the heavy chain sequence set out in SEQ ID NO: 1. Suitably, an antibody, fragment, orvariantthat may be incorporated in a pharmaceutical composition of the invention comprises a heavy chain framework region FR H2 of SEQ ID NO:

[0536] 32 or that shares at least 90%, or at least 95%, identity with such a sequence.

[0537] SEQ ID NO:33 is the framework region FR H3 of the heavy chain sequence set out in SEQ ID NO: 1. Suitably, an antibody, fragment, orvariantthat may be incorporated in a pharmaceutical composition of the invention comprises a heavy chain framework region FR H3 of SEQ ID NO:

[0538] 33 or that shares at least 90%, or at least 95%, identity with such a sequence.

[0539] SEQ ID NO:34 is the framework region FR H4 of the heavy chain sequence set out in SEQ ID NO: 1. Suitably, an antibody, fragment, orvariantthat may be incorporated in a pharmaceutical composition of the invention comprises a heavy chain framework region FR H4 of SEQ ID NO:

[0540] 34 or that shares at least 90%, or at least 95%, identity with such a sequence.

[0541] Suitably an antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may comprise a FR H1 of SEQ ID NO: 31 , a FR H2 of SEQ ID NO: 32, a FR H3 of SEQ ID NO: 33, and a FR H4 of SEQ ID NO: 34, or may share at least 90%, or at least 95%, identity with these recited sequences.

[0542] SEQ ID NO:35 is the light chain framework region FR L1 of the light chain sequence set out in SEQ ID NO: 2. In a suitable embodiment, an antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention comprises a light chain framework region FR L1 of SEQ ID NO: 35 orthat shares at least 90%, or at least 95%, identity with such a sequence.

[0543] SEQ ID NO:36 is the light chain framework region FR L2 of the light chain sequence set out in SEQ ID NO: 2. Suitably, an antibody, fragment, orvariant that may be incorporated in a pharmaceutical composition of the invention comprises a light chain framework region FR L2 of SEQ ID NO: 36 or that shares at least 90%, or at least 95%, identity with such a sequence. SEQ ID NO:37 is the light chain framework region FR L3 of the light chain sequence set out in SEQ ID NO: 2. Suitably, an antibody, fragment, orvariant that may be incorporated in a pharmaceutical composition of the invention comprises a light chain framework region FR L3 of SEQ ID NO: 37 or that shares at least 90%, or at least 95%, identity with such a sequence.

[0544] SEQ ID NO:38 is the light chain framework region FR L4 of the light chain sequence set out in SEQ ID NO: 2. Suitably, an antibody, fragment, orvariant that may be incorporated in a pharmaceutical composition of the invention comprises a light chain framework region FR L4 of SEQ ID NO: 38 or that shares at least 90%, or at least 95%, identity with such a sequence.

[0545] Suitably an antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may comprise a FR L1 of SEQ ID NO: 35, a FR L2 of SEQ ID NO: 36, a FR L3 of SEQ ID NO: 37, and a FR L4 of SEQ ID NO: 38, or may share at least 90%, or at least 95%, identity with these recited sequences.

[0546] In a suitable embodiment, an antibody, fragment, or variant that may be incorporated in a pharmaceutical composition of the invention may comprise a FR H1 of SEQ ID NO: 31 , a FR H2 of SEQ ID NO: 32, a FR H3 of SEQ ID NO: 33, a FR H4 of SEQ ID NO: 34, a FR L1 of SEQ ID NO: 35, a FR L2 of SEQ ID NO: 36, a FR L3 of SEQ ID NO: 37, and a FR L4 of SEQ ID NO: 38, or may share at least 90%, or at least 95%, identity with these recited sequences.

[0547] The exemplary sequences of these framework regions set out in the sequence information table have been calculated using a combination of the Kabat and Chothia numbering systems. It will be appreciated that the definitions of the framework regions may alter slightly based upon alternative numbering systems. Such changes will correspond to changes in the definitions of the CDRs determined by these alternative numbering systems (as set out in Table 69), and so it may be preferred to use numbering systems consistently to determine both CDRs and framework regions.

[0548] Variants of SelK2 antibodies

[0549] For the purposes of the present disclosure, a variant of an antibody refers to a polypeptide that has an amino acid sequence that comprises at least one alteration as compared to the amino acid sequence of SelK2 (the antibody consisting of the heavy and light chains of SEQ ID NO: 1 and SEQ ID NO: 2). An “alteration” as used herein refers to any change made to the amino acid sequence of a polypeptide such that its sequence is not identical to that of the corresponding sequence of a SelK2 antibody. Alterations may, for example, comprise deletion of one or more amino acid residues found in the sequence of an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2, or addition of one or more amino acid resides not found in the sequence of an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2. Alterations may comprise substitutions of one or more amino acid residues with residues that do not correspond to those present in the sequence of an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2.

[0550] One or more different types of alterations (for example, additions, deletions or substitutions) may be present in an amino acid sequence of a variant of SelK2.

[0551] Thus, a variant of an antibody is polypeptide with an amino acid sequence that shares at least 90% identity, but less than 100% identity, with the sequence of a SelK2 antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2. For example, a suitable variant may share at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or more, identity with the sequence of an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2. However, a variant of an antibody suitable for incorporation in a pharmaceutical composition of the invention will not share 100% identity with an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2.

[0552] The amino acid sequence of a variant of an antibody may comprise a single alteration as compared to the corresponding sequence of an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2. Alternatively, a variant of an antibody may have an amino acid sequence that comprises at least 1 alteration as compared to the corresponding sequence of an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2. For example, a variant of an antibody may have an amino acid sequence that comprises up to 5 alterations as compared to the corresponding sequence of an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2, for example comprising up to 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, or 85 alterations as compared to the sequence of SelK2.

[0553] For example, a variant of an antibody may have an amino acid sequence that comprises 2, 3, 4, 5, 6, 7, 8, 9 or 10 alterations as compared to the corresponding sequence of an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2. A variant of an antibody may have an amino acid sequence that comprises 1 1 , 12, 13, 14, 15, 16, 17, 18, 19 or 20 alterations as compared to the corresponding sequence of an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2. A variant of an antibody may have an amino acid sequence that comprises 21 , 22, 23, 24, 25, 26, 27, 28, 29 or 30 alterations as compared to the corresponding sequence of an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2. A variant of an antibody may have an amino acid sequence that comprises 31 , 32, 33, 34, 35, 36, 37, 38, 39 or 40 alterations as compared to the corresponding sequence of an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2. A variant of an antibody may have an amino acid sequence that comprises 41 , 42, 43, 44, 45, 46, 47, 48, 49 or 50 alterations as compared to the corresponding sequence of an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2. A variant of an antibody may have an amino acid sequence that comprises 51 , 52, 53, 54, 55, 56, 57, 58, 59 or 60 alterations as compared to the corresponding sequence of an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2. A variant of an antibody may have an amino acid sequence that comprises 61 , 62, 63, 64, 65, 66, 67, 68, 69 or 70 alterations as compared to the corresponding sequence of an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2. A variant of an antibody may have an amino acid sequence that comprises 71 , 72, 73, 74, 75, 76, 77, 78, 79 or 80 alterations as compared to the corresponding sequence of an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2. A variant of an antibody may have an amino acid sequence that comprises 81 , 82, 83, 84, 85, or86 alterations as compared tothe corresponding sequenceof an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2.

[0554] Suitably, a variant of an antibody may have an amino acid sequence that comprises at least 2 alterations, at least 3 alterations, at least 4 alterations, at least 5 alterations, at least 6 alterations, at least 7 alterations, at least 8 alterations, at least 9 alterations, at least 10 alterations, at least 15 alterations, at least 20 alterations, at least 25 alterations, at least 30 alterations, at least 35 alterations, at least 40 alterations, at least 45 alterations, at least 50 alterations, at least 55 alterations, at least 60 alterations, at least 65 alterations, at least 70 alterations, at Ieast75 alterations, at least 80 alterations, orat least 85 alterations as compared to the corresponding sequence of an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2.

[0555] In a suitable embodiment, a variant antibody suitable for incorporation in a pharmaceutical composition of the invention may comprise a heavy chain with an amino acid sequence that comprises no more than 10 amino acid alterations as compared to the sequence set out in SEQ ID NO: 1. For example, a variant antibody suitable for incorporation in a pharmaceutical composition of the invention may comprise a heavy chain with an amino acid sequence that comprises no more than 9, no more than 8, no more than 7, no more than 6, no more than 5, no more than 4, no more than 3, no more than 2, or no more than 1 amino acid alteration as compared to the sequence set out in SEQ ID NO: 1 . In a suitable embodiment, a variant antibody suitable for incorporation in a pharmaceutical composition of the invention may comprise a light chain with an amino acid sequence that comprises no more than 10 amino acid alterations as compared to the sequence set out in SEQ ID NO: 2. For example, a variant may comprise a light chain with an amino acid sequence that comprises no more than 9, no more than 8, no more than 7, no more than 6, no more than 5, no more than 4, no more than 3, no more than 2, or no more than 1 amino acid alteration as compared to the sequence set out in SEQ ID NO: 2.

[0556] In a suitable embodiment, a variant antibody suitable for incorporation in a pharmaceutical composition of the invention may comprise a heavy chain that shares at least 80% identity with the heavy chain sequence set out in SEQ ID NO: 1. For example, a variant may comprise a heavy chain that shares at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, least 93%, at least 94%, least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the heavy chain sequence set out in SEQ ID NO: 1 . In such an embodiment the light chain sequence of the variant antibody may comprise or consist of that set out in SEQ ID NO: 2, or may also comprise one or more alterations as compared to the reference sequence.

[0557] Alternatively, or additionally, a variant antibody suitable for incorporation in a pharmaceutical composition of the invention may comprise a light chain that shares at least 80% identity with the light chain sequence set out in SEQ ID NO: 2. Forexample, a variant may comprise a light chain that shares at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the light chain sequence set out in SEQ ID NO: 2. Suitably the heavy chain sequence of the variant antibody may comprise or consist of that set out in SEQ ID NO: 1 , or may also comprise one or more alterations as compared to the reference sequence.

[0558] In a suitable embodiment, a variant antibody suitable for incorporation in a pharmaceutical composition of the invention may comprise a heavy chain that shares at least 80% identity with the heavy chain sequence set out in SEQ ID NO: 1 and a light chain that shares at least 80% identity with the light chain sequence setoutin SEQ ID NO: 2. In a suitable embodiment, such a variant of an antibody shares at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with both the heavy chain sequence set out in SEQ ID NO: 1 and with the light chain sequence set out in SEQ ID NO: 2.

[0559] In a suitable embodiment, a variant antibody suitable for incorporation in a pharmaceutical composition of the invention may comprise a heavy chain variable region with an amino add sequence that comprises no more than 10 amino acid alterations as compared to the sequence set out in SEQ ID NO: 3. For example, a variant antibody suitable for incorporation in a pharmaceutical composition of the invention may comprise a heavy chain variable region with an amino acid sequence that comprises no more than 9, no more than 8, no more than 7, no more than 6, no more than 5, no more than 4, no more than 3, no more than 2, or no more than 1 amino acid alteration as compared to the sequence set out in SEQ ID NO: 3.

[0560] In a suitable embodiment, a variant antibody suitable for incorporation in a pharmaceutical composition of the invention may comprise a light chain variable region with an amino acid sequence that comprises no more than 10 amino acid alterations as compared to the sequence set out in SEQ ID NO: 4. For example, a variant antibody suitable for incorporation in a pharmaceutical composition of the invention may comprise a light chain variable region with an amino acid sequence that comprises no more than 9, no more than 8, no more than 7, no more than 6, no more than 5, no more than 4, no more than 3, no more than 2, or no more than 1 amino acid alteration as compared to the sequence set out in SEQ ID NO: 4.

[0561] In a suitable embodiment, a variant antibody suitable for incorporation in a pharmaceutical composition of the invention may comprise a heavy chain variable region that shares at least 80% identity with the heavy chain variable sequence set out in SEQ ID NO: 3. For example, a variant may comprise a heavy chain variable region that shares at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the heavy chain sequence set out in SEQ ID NO: 3. In such an embodiment the light chain variable sequence of the variant of an antibody may comprise or consist of that set out in SEQ ID NO: 4, or may also comprise one or more alterations as compared to the reference sequence.

[0562] Alternatively, or additionally, a variant antibody suitable for incorporation in a pharmaceutical composition of the invention may comprise a light chain variable region that shares at least 80% identity with the light chain variable region sequence set out in SEQ ID NO: 4. For example, a variant may comprise a light chain variable region that shares at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the light chain variable sequence set out in SEQ ID NO: 4. Suitably the heavy chain variable sequence of such a variant of an antibody may comprise or consist of that set out in SEQ ID NO: 3, or may also comprise one or more alterations as compared to the reference sequence. In a suitable embodiment, a variant antibody suitable for incorporation in a pharmaceutical composition of the invention may comprise both heavy and light chain variable regions that respectively share at least 80% identity with the heavy chain variable sequence set out in SEQ ID NO: 3 and at least 80% identity with the light chain variable sequence set out in SEQ ID NO: 4. In a suitable embodiment, a variant may comprise heavy and light chain variable regions that respectively share at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the heavy chain variable sequence set out in SEQ ID NO: 3 and with the light chain variable sequence set outin SEQ ID NO: 4. In a suitable embodiment, a variant may comprise a heavy chain variable region sharing at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 3 and a light chain variable region sharing at least 95% sequence identity with the amino acid sequence of SEQ ID NO: 4.

[0563] In a suitable embodiment, a variant antibody suitable for incorporation in a pharmaceutical composition of the invention may comprise a heavy chain constant region with an amino add sequence that comprises no more than 10 amino acid alterations as compared to the sequence set out in SEQ ID NO: 5. For example, a variant may comprise a heavy chain constant region with an amino acid sequence that comprises no more than 9, no more than 8, no more than 7, no more than 6, no more than 5, no more than 4, no more than 3, no more than 2, or no more than 1 amino acid alteration as compared to the sequence set out in SEQ ID NO: 5.

[0564] In a suitable embodiment, a variant antibody suitable for incorporation in a pharmaceutical composition of the invention may comprise a heavy chain constant region sharing at least 80% sequence identity with SEQ ID NO: 5. For example, a variant may comprise a heavy chain constant region sharing at least 80%, at least 85%, at least 90%, at least 91 %, least 92%, at least 93%, least 94%, at least 95%, least 96%, at least 97%, least 98%, or at least 99% identity with SEQ ID NO: 5.

[0565] In a suitable embodiment, a variant antibody suitable for incorporation in a pharmaceutical composition of the invention may comprise a light chain constant region with an amino acid sequence that comprises no more than 10 amino acid alterations as compared to the sequence set out in SEQ ID NO: 6. For exam pie, a variant may comprise a light chain constant region with an amino acid sequence that comprises no more than 9, no more than 8, no more than 7, no more than 6, no more than 5, no more than 4, no more than 3, no more than 2, or no more than 1 amino acid alteration as compared to the sequence set out in SEQ ID NO: 6. In a suitable embodiment, a variant antibody suitable for incorporation in a pharmaceutical composition of the invention may comprise a light chain constant region sharing at least 80% sequence identity with SEQ ID NO: 6. For example, a variant may comprise a light chain constant region sharing at least 80%, at least 85%, at least 90%, at least 91 %, least 92%, at least 93%, least 94%, at least 95%, least 96%, at least 97%, least 98%, or at least 99% identity with SEQ ID NO: 6. In a suitable embodiment, a SelK2 antibody of the invention may comprise a light chain constant region of SEQ ID NO: 6.

[0566] In a suitable embodiment, a variant antibody suitable for incorporation in a pharmaceutical composition of the invention may comprise both heavy and light chain constant regions that respectively share at least 80% identity with the heavy chain constant region set out in SEQ ID NO: 5 and at least 80% identity with the light chain constant region set out in SEQ ID NO: 6. In a suitable embodiment, a variant may comprise heavy and light chain constant regions that respectively share at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the heavy chain constant sequence set out in SEQ ID NO: 5 and with the light chain constant sequence set out in SEQ ID NO: 6.

[0567] In a suitable embodiment, alterations present in an antibody variant incorporated in a pharmaceutical composition of the invention are limited to the constant regions of the sequence.

[0568] Fragments of SelK2 antibodies

[0569] It will be appreciated that a fragment of an antibody comprising or consisting of SEQ I D NO: 1 and SEQ ID NO: 2 refers to a polypeptide that shares 100% sequence identity with the reference sequence over its length, but that is truncated in comparison to the full length antibody.

[0570] Thus, a fragment may comprise at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, or more, of the full length of the corresponding SelK2 antibody. Suitably, a fragment may comprise at least 96%, at least 97%, at least 98%, at least 99% or more, of the full length of the corresponding SelK2 antibody.

[0571] In a suitable embodiment, a fragment of an antibody may have an amino acid sequence that comprises 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acid residues lost by truncation as compared to the corresponding sequence of an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2. A fragment of an antibody may have an amino acid sequence that comprises 11 , 12, 13, 14, 15, 16, 17, 18, 19 or 20 amino acid residues lost by truncation as compared to the corresponding sequence of an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2. A fragment of an antibody may have an amino acid sequence that comprises 21 , 22, 23, 24, 25, 26, 27, 28, 29 or 30 amino acid residues lost by truncation as compared to the corresponding sequence of an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2. A fragment of an antibody may have an amino acid sequence that comprises 31 , 32, 33, 34, 35, 36, 37, 38, 39 or 40 amino acid residues lost by truncation as compared to the corresponding sequence of an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2. A fragment of an antibody may have an amino acid sequence that comprises 41 , 42, 43, 44, 45, 46, 47, 48, 49 or 50 amino acid residues lost by truncation as compared to the corresponding sequence of an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2. A fragment of an antibody may have an amino acid sequence that comprises 51 , 52, 53, 54, 55, 56, 57, 58, 59 or 60 amino acid residues lost by truncation as compared to the corresponding sequence of an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2. A fragment of an antibody may have an amino acid sequence that comprises 61 , 62, 63, 64, 65, 66, 67, 68, 69 or 70 amino acid residues lost by truncation as compared to the corresponding sequence of an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2. A fragment of an antibody may have an amino acid sequence that comprises 71 , 72, 73, 74, 75, 76, 77, 78, 79 or 80 amino acid residues lost by truncation as compared to the corresponding sequence of an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2. A fragment of an antibody may have an amino acid sequence that comprises 81 , 82, 83, 84, 85, or 86 amino acid residues lost by truncation as compared to the corresponding sequence of an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2.

[0572] Suitably, a fragment of a SelK2 antibody suitable for incorporation in a pharmaceutical composition of the invention may lack no more than 1 residue as compared to an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2. Alternatively, a fragment may lack no more than 2 residues, 3 residues, 4 residues, 5 residues, 6 residues, 7 residues, 8 residues, 9 residues, or 10 residues of the full length Selk2 antibody. Indeed, a suitable fragment may lack no more than 12 residues, 14 residues, 16 residues, 18 residues or no more than 20 residues of the full length SelK2 antibody. A suitable fragment may lack no more than 25 residues, 30 residues, 35 residues, 40 residues, 45 residues, 50 residues, 55 residues, 60 residues, 65 residues, 75 residues, 80 residues, or 85 residues of the full length SelK2 antibody.

[0573] In a suitable embodiment, an antibody fragment suitable for incorporation in a pharmaceutical composition of the invention may comprise a heavy chain with an amino acid sequence that lacks no more than 10 amino acid residues as compared to the sequence set out in SEQ ID NO: 1 . For example, a fragment may comprise a heavy chain with an amino acid sequence that lacks no more than 9, no more than 8, no more than 7, no more than 6, no more than 5, no more than 4, no more than 3, no more than 2, or no more than 1 amino acid residue as compared to the sequence set out in SEQ ID NO: 1 .

[0574] In a suitable embodiment, an antibody fragment suitable for incorporation in a pharmaceutical composition of the invention may comprise a light chain with an amino acid sequence that lacks no more than 10 amino acid residues as compared to the sequence set out in SEQ ID NO: 2. For example, a fragment may comprise a light chain with an amino acid sequence that lacks no more than 9, no more than 8, no more than 7, no more than 6, no more than 5, no more than 4, no more than 3, no more than 2, or no more than 1 amino acid residue as compared to the sequence set out in SEQ ID NO: 2.

[0575] In a suitable embodiment, an antibody fragment suitable for incorporation in a pharmaceutical composition of the invention may comprise a heavy chain that comprises at least 80% of the full length heavy chain sequence set out in SEQ ID NO: 1 . For example, a fragment may comprise a heavy chain that comprises at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, least 93%, at least 94%, least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the full length heavy chain sequence set out in SEQ ID NO: 1 . In such an embodiment the light chain sequence of the fragment of the antibody may comprise or consist of that set out in SEQ ID NO: 2, or may also comprise one or more truncated residues as compared to the reference sequence.

[0576] Alternatively, or additionally, an antibody fragment suitable for incorporation in a pharmaceutical composition of the invention may comprise a light chain that comprises at least 80% of the full length light chain sequence set out in SEQ ID NO: 2. For example, a fragment may comprise a light chain that comprises at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the full length light chain sequence set out in SEQ ID NO: 2. Suitably the heavy chain sequence of such afragment of an antibody may comprise or consist of that set out in SEQ ID NO: 1 , or may also comprise one or more truncated residues as compared to the reference sequence.

[0577] In a suitable embodiment, an antibody fragment suitable for incorporation in a pharmaceutical composition of the invention may comprise a heavy chain that comprises at least 80% of the full length heavy chain sequence set out in SEQ ID NO: 1 and a light chain that comprises at least 80% of the full length light chain sequence set out in SEQ ID NO: 2. In a suitable embodiment, such a fragment of an antibody comprises at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of both the full length heavy chain sequence set out in SEQ ID NO: 1 and the full length light chain sequence set out in SEQ ID NO: 2.

[0578] In a suitable embodiment, an antibody fragment suitable for incorporation in a pharmaceutical composition of the invention may comprise a heavy chain variable region with an amino add sequence that lacks no more than 10 amino acid residues as compared to the sequence set out in SEQ ID NO: 3. For example, a fragment may comprise a heavy chain variable region with an amino acid sequence that lacks no more than 9, no more than 8, no more than 7, no more than 6, no more than 5, no more than 4, no more than 3, no more than 2, or no more than 1 amino acid residue as compared to the sequence set out in SEQ ID NO: 3.

[0579] In a suitable embodiment, an antibody fragment suitable for incorporation in a pharmaceutical composition of the invention may comprise a light chain variable region with an amino acid sequence that lacks no more than 10 amino acid residues as compared to the sequence set out in SEQ ID NO: 4. For example, a fragment may comprise a light chain variable region with an amino acid sequence that lacks no more than 9, no more than 8, no more than 7, no more than 6, no more than 5, no more than 4, no more than 3, no more than 2, or no more than 1 amino acid residue as compared to the sequence set out in SEQ ID NO: 4.

[0580] In a suitable embodiment, an antibody fragment suitable for incorporation in a pharmaceutical composition of the invention may comprise a heavy chain variable region that comprises at least 80% identity of the full length heavy chain variable sequence set out in SEQ ID NO: 3. For example, a may comprise a heavy chain variable region that comprises at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity of the full length heavy chain sequence set out in SEQ ID NO: 3. In such an embodiment the light chain variable sequence of the fragment of an antibody may comprise or consist of that set out in SEQ ID NO: 4, or may also comprise one or more truncated residues as compared to the reference sequence.

[0581] Alternatively, or additionally, an antibody fragment suitable for incorporation in a pharmaceutical composition of the invention may comprise a light chain variable region that comprises at least 80% identity of the full length light chain variable region sequence set out in SEQ ID NO: 4. For example, a fragment may comprise a light chain variable region that comprises at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the full length light chain variable sequence set out in SEQ ID NO: 4. Suitably the heavy chain variable sequence of such a fragment of an antibody may comprise or consist of that set out in SEQ ID NO: 3, or may also comprise one or more truncated residues as compared to the reference sequence.

[0582] In a suitable embodiment, an antibody fragment suitable for incorporation in a pharmaceutical composition of the invention may comprise both heavy and light chain variable regions that respectively comprise at least 80% of the full length heavy chain variable sequence set out in SEQ ID NO: 3 and at least 80% of the full length light chain variable sequence set out in SEQ ID NO: 4. In a suitable embodiment, a fragment may comprise heavy and light chain variable regions that respectively comprise at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the full length heavy chain variable sequence set out in SEQ ID NO: 3 and of the full length light chain variable sequence set out in SEQ ID NO: 4. In a suitable embodiment, an antibody fragment suitable for incorporation in a pharmaceutical composition of the invention may comprise a heavy chain variable region comprising at least 95% of the full length heavy chain variable region of SEQ ID NO: 3 and a light chain variable region comprising at least 95% of the full length light chain variable region of SEQ ID NO: 4.

[0583] In a suitable embodiment, an antibody fragment suitable for incorporation in a pharmaceutical composition of the invention may comprise a heavy chain constant region with an amino add sequence that lacks no more than 10 amino acid residues as compared to the sequence set out in SEQ ID NO: 5. For example, a fragment may comprise a heavy chain constant region with an amino acid sequence that lacks no more than 9, no more than 8, no more than 7, no more than 6, no more than 5, no more than 4, no more than 3, no more than 2, or no more than 1 amino acid residue as compared to the sequence set out in SEQ ID NO: 5.

[0584] In a suitable embodiment, an antibody fragment suitable for incorporation in a pharmaceutical composition of the invention may comprise a heavy chain constant region comprising at least 80% of the full length amino acid sequence of SEQ ID NO: 5. For example, a fragment may comprise a heavy chain constant region comprising at least 80%, at least 85%, at least 90%, at least 91 %, least 92%, at least 93%, least 94%, at least 95%, least 96%, at least 97%, least 98%, or at least 99% of the full length amino acid sequence of SEQ ID NO: 5.

[0585] In a suitable embodiment, an antibody fragment suitable for incorporation in a pharmaceutical composition of the invention may comprise a light chain constant region with an amino acid sequence that lacks no more than 10 amino acid residues as compared to the sequence set out in SEQ ID NO: 6. For example, a fragment may comprise a light chain constant region with an amino acid sequence that lacks no more than 9, no more than 8, no more than 7, no more than 6, no more than 5, no more than 4, no more than 3, no more than 2, or no more than 1 amino acid residue as compared to the sequence set out in SEQ ID NO: 6.

[0586] In a suitable embodiment, an antibody fragment suitable for incorporation in a pharmaceutical composition of the invention may comprise a light chain constant region comprising at least 80% of the full length amino sequence of SEQ ID NO: 6. For example, a fragment may comprise a light chain constant region comprising at least 80%, at least 85%, at least 90%, at least 91 %, least 92%, at least 93%, least 94%, at least 95%, least 96%, at least 97%, least 98%, or at least 99% of the full length amino aacis sequence of SEQ ID NO: 6.

[0587] In a suitable embodiment, an antibody fragment suitable for incorporation in a pharmaceutical composition of the invention may comprise both heavy and light chain constant regions that respectively comprise at least 80% of the full length heavy chain constant region set out in SEQ ID NO: 5 and at least 80% of the full length light chain constant region set out in SEQ ID NO: 6. In a suitable embodiment, a fragment may comprise both heavy and light chain constant regions that respectively share at least 80%, at least 85%, at least 90%, at Ieast 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of both the full length heavy chain constant sequence set out in SEQ ID NO: 5 and the full length light chain constant sequence set out in SEQ ID NO: 6.

[0588] Post-translational modifications

[0589] Suitably an antibody suitable for incorporation in a pharmaceutical composition of the invention may comprise one or more post-translational modifications. In a suitable embodiment, the one or more post-translational modifications may be selected from the group consisting of: glycosylation; N-terminal glutaminyl cyclization; and C-terminal lysine clipping.

[0590] In a suitable embodiment, an antibody, antibody fragment, or variant, suitablefor incorporation in a pharmaceutical composition of the invention has a heavy chain that is glycosylated atAsn- 289 with G0F as the major N-glycan structure.

[0591] Methods of manufacturing In a fifth aspect, the invention provides a method for the preparation of a pharmaceutical composition comprising approximately 210 mg / mL SelK2 or a fragment or variant thereof, the method comprising combining:

[0592] • a solution comprising an antibody at a concentration of greater than 210 mg / ml in 10mM acetate, wherein the antibody comprises a heavy chain of SEQ ID NO: 1 and an antibody light chain of SEQ ID NO: 2, or a fragment or variant thereof sharing at least 90% sequence identity with SelK2; with

[0593] • 110mM sorbitol; 0.1 % poloxamer 188; 50mM proline; and 110 mM mannitol;

[0594] • to form a mixture comprising the antibody at a concentration of approximately 210 mg / mL; and

[0595] • sterilizing the mixture, to produce the pharmaceutical composition.

[0596] In a suitable embodiment, the method in accordance with the fifth aspect of the invention further comprises adjusting the pH to 5.3 using 1 % acetic acid. Suitably the sterilizing is by filtration, for example by using a 0.2 μm PES filter.

[0597] Further definitions

[0598] As used herein the term “about” when used in connection with a referenced numeric indication means the referenced numeric indication plus or minus up to 5% of that referenced numeric indication, unless otherwise specifically providedfor herein. Forexample, the language “about 50%” covers the range of 45% to 55%. In various embodiments, the term “about” when used in connection with a referenced numeric indication can mean the referenced numeric indication plus or minus up to 4%, 3%, 2%, 1 %. 0.5%, or 0.25% of that referenced numeric indication (unless specifically provided for in the claims).

[0599] EXAMPLES

[0600] Example 1. Affinity of the parental antibody and SelK2

[0601] SelK2 and the parental antibody were immobilized and their interaction with SelSP1 (PSGL-1 sulfopeptide) was characterized using SensiQ Pioneer.

[0602] Materials and Methods

[0603] SensiQ Pioneer (S / N 131 10154) was used for all SPR measurements at a controlled temperature of 25°C. SelK2 (Lot # 14-2189) was dispensed into three 1 mL aliquots in the biosafety cabinet. Protein G was purchased from BioVision (Cat. # 6510). Running bufferfor Pioneer contained 10 mM HEPES pH 7.4, 150 mM NaCI, 3.4 mM EDTA and 0.005% Tween- 20. All other reagents were purchased from Sigma Aldrich.

[0604] A new COOH2 biosensor was installed and conditioned according to CH0012 protocol. Protein G was immobilized via amine coupling. A mixture containing 0.1 M EDC and 0.025M NHS was injected for 4 minutes overflow channel 1 (FC1 ) and FC2. 50 μg / mL Protein G in 10 mM Sodium acetate buffer pH 4.5 was injected for 15 minutes over FC1 and FC2. 1 M Ethanolamine HCI solution pH 8.0 was injected for 5 minutes over FC1 and FC2. Approximately 1 ,145 response units (RU) of Protein G was coupled on FC1 and FC2 of the sensor chip. 50 nM SelK2 in system buffer was injected to capture approximately 2,000 RU SelK2 on FC1 and 1 ,000 RU on FC2. A dilution series of SelSP1 was prepared in system buffer 50, 25, 12.5, 6.25, and 3.125 nM. Each sample was injected for 3 min at a flow rate of 40 μL / min. Dissociation was monitored after each injection for 4 min at the same flow rate. Samples were tested in duplicate from low concentration to high.

[0605] Kinetic model fitting was performed using Qdat analysis software (SensiQ Technologies, Inc. Version 2.5.1.56). A simple 1 : 1 kinetic model was fit to the data to determine best fit values for association and dissociation rate constants, ka and kd. A local Rmax was fit for each curve to account for incomplete dissociation of the SelSP1 after some of the analyte injections. Global analysis of the binding responses revealed good agreement in kinetics and affinity between the 2,000 (high) RU and 1 ,000 (low) RU SelK2 surfaces. T able 2 shows the kinetic rate and affinity constants for this lot of SelK2.

[0606] Results Table 1 - Kinetic rate constants of the parental and SelK2 antibody binding SelSP1

[0607] As shown in FIG. 1 and Table 1 , the parental antibody binds SelSP1 with an equilibrium dissociation constant (KD) of 16.4 nM. As shown in FIGS. 2, FIGS. 3, and Table 4, the SelK2 antibody binds SelSP1 with an equilibrium dissociation constant (KD) of 4.28 nM. This result was consistent when tested at two surface densities of the antibody and using two replicates for all SelSP1 concentrations. Thus, surprisingly the SelK2 antibody has improved binding affinity to SelSP1 in comparison to the parental antibody.

[0608] Example 2. SelK2 can functionally block neutrophil rolling on P-selectin.

[0609] In vitro rolling assay

[0610] The ability of SelK2 to functionally block PSGL-1 was tested by assessing its ability to block neutrophil rolling on P-selectin. In this assay, human neutrophils roll and tether to P-selectin coated on a plate at a density similar to that found on activated endothelial cells underflow conditions that simulate sheer stress in blood vessels.

[0611] Materials and Methods

[0612] P-selectin purified from platelet membranes (mP-selectin) was prepared as described (Ushiyama et al., 1993). The parental antibody was purchased from Millipore (catalog # MAB4092). Human neutrophils were isolated from healthy donors as described (Zimmerman et al., 1985). A 30 μl drop of human mP-selectin (1 μg / ml) was placed in a demarcated area on a 35-mm culture plate (Corning) and incubated at 4°C overnight. The area was washed twice with HBSS and then blocked with HBSS containing 1 % human serum albumin at room temperature for 2 hours. Human neutrophils (4.5 x 105 / ml in HBSS containing 0.5% human serum albumin) were incubated with buffer alone (control) or SelK2 at concentrations of 1 μg / mL, 0.5 μg / mL, 0.25 μg / mL, 0.125 μg / mL, 0.06 μg / mL, 0.03 μg / mL, or 0.015 μg / mL for 5 minutes before the rolling experiments started. The cells were then perfused over the adsorbed mP-selectin in a parallel-plate flow chamber under shear stress at 1 .0 dyn / cm2(Ramachandran et al. , 1999). After 5 minutes, dynamic images of several fields of view were captured and digitalized by a HAMAMATSU ORCA-Flash2.8 CCD digital camera. The accumulated number of bound and slow-rolling neutrophils from each run (SelK2 concentration) was measured in 5 to 6 different fields by image analysis software Element (Nikon) to calculate the average cell number and standard deviation.

[0613] Table 2. Neutrophil inhibition bySelK2.

[0614] SelK2 inhibited neutrophil rolling in this assay in a dose dependent manner over a concentration range of 0.125 μg / mL to 1 μg / mL, achieving near complete inhibition at 0.5 μg / mL, as shown in FIG. 4 and T able 2. These results demonstrate the ability of the SelK2 antibody to block PSGL-1 and inhibit neutrophil binding to P-selectin and rolling under shear stress.

[0615] Example 3. SelK2 can functionally block and inhibit chemokine CCL27 binding and interacting with PSGL-1

[0616] The ability of SelK2 to block and inhibit chemokine CCL27 binding and interacting with PSGL- 1 was assessed using an SPR-based assay. The assay utilizes a sulfopeptide (SelSP1) that is modelled after the N-terminus of human PSGL-1 , and which interacts with CCL27 and is bound by K2 and SelK2.

[0617] Materials and Methods

[0618] All reagents were purchased from Sigma Aldrich (St. Louis, MO) unless otherwise noted. SensiQ Pioneer was used for all SPR experiments. CCL27 (376-CT-025 / CF)was supplied by Selexys from R&D Systems. SelSP1 , K2, and SelK2 were also supplied by Selexys. Assay buffercontained 10 mM HEPES pH 7.4, 150 mM NaCI, 2 mM CaCI2 and 0.01 % (w / w) Tween- 20. Buffers were sterile filtered prior to use.

[0619] Sensor Preparation: A new COOH2 sensor was installed and was conditioned by two 10 second injections of each of the following: 10 mM HCI, 50 mM NaOH, and 0.1 % SDS. The instrument was primed in assay buffer. A solution containing 75 μg / mL Streptavidin, 4 mM EDC and 1 mM NHS in a 10 mM Acetate buffer pH 4.5 was injected over all three flow channels (FC) until ~3000 RU of protein was captured. The sensor was deactivated by injecting 1 M Ethanolamine pH 8.0 for 4 min to cap unreacted NHS esters. A solution containing 500 nM SelSP1 in assay buffer was injected over FC3 and FC2 for 3 min. Approximately 300 RU were captured on FC2 and 250 RU were captured on FC3. FC1 was blocked by injecting 13 pM biotin-PEO4-NH2 for 2 min. The system was primed again in assay buffer.

[0620] CCL27 Binding Assay: The CCL27 binding assay consisted of four injections where the first injectwas eitherbuffer, 40 μg / mL K2 or40 μg / mLSelK2; the second injectwas 990 nM CCL27 in assay buffer; and the third and fourth injects were 0.1 % SDS and 20 mM NaOH. Inject 1 was for20 minute, inject2 was 2 minutes and injects3 and 4 were each 1 minute. Injects were flowed over all three FCs. Each assay cycle was repeated three times in sequential order.

[0621] Data Analysis: Data were processed and analysed using Qdat (BioLogic Software & SensiQ Technologies, Inc.). Assay curves were overlaid on the X axis and the Y axis was normalized at a time point directly before the CCL27 inject. The reference channel signal was subtracted from the SelSP1 channel signal to subtract non-specific artifacts. The identity of the blocking injection sample was used to differentiate the assay cycles and compare the CCL27 binding. The blocking injection samples were alternated, and each was repeated in triplicate. The replicates of each were compared together because an overall decrease in CCL27 binding was observed for each of the three blocking samples. So the Ab blocked CCL27 injects were referenced against the buffer blocked cycle fortheir respective replicate set. Percent inhibition was calculated using the CCL27 injection after buffer inject as a control for each Ab blocked replicate.

[0622] Results

[0623] FIG. 5 shows the complete assay cycles for each blocking sample where the parental and SelK2 cycles (dot-dash line and dashed line curves, respectively) show large binding responses of antibody in injection ‘A while the solid line curves have negligible binding in this inject. FIG. 6 shows the same data zoomed into injection ‘B’ and re-normalized just before this injection. This view illustrates that the CCL27 binds SelSP1 with weak affinity (KD 10pM, fit not shown) in the absence of antibody blocking (buffer control). This binding is decreased when the SelSP1 is blocked with either of the antibodies. Table 3. Percent CCL27 Inhibition

[0624] Table 3 shows the percent inhibition for the parental and SelK2 antibodies at each replicate and on two SelSP1 reaction FCs. SelK2 was found to inhibit CCL27 binding by 62 ±6%. The parental antibody was found to inhibit CCL27 binding by 50 ±5%. The slight increase in inhibition of SelK2 over K2 is most likely due to its ability to bind more to SelSP1 than K2. These results demonstrate the ability of the SelK2 antibody to block chemokine binding through PSGL-1 inhibition.

[0625] The inventors submit that this characteristic of SelK2’s binding can be linked either to a conformation of binding which allows for better packing density of antibody or higher affinity toward the SelSP1 target.

[0626] Thus, this study demonstrates that these two antibodies which bind to SelSP1 can specifically block CCL27 binding to SelSP1 , indicating that the CCL27 recognizes a site on SelSP1 which overlaps with its antibody binding sites. The affinity of both antibodies for SelSP1 is several orders of magnitude stronger than CCL27 and therefore, the likelihood of antibody displacing CCL27 in vivo is very high.

[0627] T aken together, the data of Examples 2 and 3 demonstrate PSGL-1 binding affinity, specificity, and functionality of the SelK2 antibody. These results also demonstrate a dual function for SelK2. The N-terminal domain of PSGL-1 contains a selectin binding domain that overlaps with a chemokine-binding domain. SelK2 binds to this domain and effectively blocks both selectin and chemokine binding. Example 4. SelK2 Effectively Inhibits Human Neutrophil Binding to P-selectin Under Flow

[0628] To determine the effectiveness of SelK2 in preventing the adhesive interactions between PSGL-1 and its main ligand P-selectin, human neutrophils were incubated with SelK2 prior to perfusing the cells over P-selectin coated plates under physiologic flow.

[0629] Human neutrophils were incubated with buffer alone (Control) or SelK2 at concentrations of 0.5, 0.25 or 0.125 μg / mL for 5 minutes. The cells were then perfused over a surface coated with human P-selectin in a parallel-plate flow chamber under a shear stress of 1 .0 dyn / cm2. Dynamic images of representative fields were recorded using a digital camera. Elongated streaks depict cells initially rolling on the surface of plate bound P-selectin while rounded cells represent cells that have transiently adhered to P-selectin.

[0630] As shown in FIG. 7, SelK2 inhibited neutrophil rolling on P-selectin in a dose dependent manner over a concentration range of 0.125 to 0.5 μg / mL, achieving near complete inhibition at 0.5 μg / mL.

[0631] Example 5. Pharmacokinetic Analysis

[0632] PK ELISA Assay

[0633] A PK ELISA assay was used to measure the concentration of SelK2 determined by a ligand binding antigen ELISA (LBA-ELISA) method. Following administration of 7.5 mg / kg Selk2 to subjects on Visit 4 Day 1 (Dose 1 ) and Visit 8 Day 22 (Dose 2), serum samples were collected for SelK2 analysis at time points: 15 minutes, 30 minutes, 1 hour, 24 hours, 8 days, 15 days, 29 days, 36 days, 43 days, 50 days, and 57 days post-administration, togetherwith a pre-dose collection.

[0634] Neutravidin coated 96-well ELISA plates were loaded with biotinylated SP-1 peptide and diluted serum samples were then loaded and the SelK2 was captured. The SelK2 was then detected with a goat anti-human IgG that is HRP conjugated and a colorimetricsignal can then be generated by developing with a TMB substrate. The concentration of SelK2 in serum samples is determined by measuring the sample signal to the signal of a known calibration curve containing multiple concentrations of SelK2. As shown in FIG. 8 and Table 4, the mean and medium half-life is 322 and 268 hours, respectively. In addition, the mean and medium time of maximum observed serum concentration for dose 1 was 1.97 hours and 0.9 hours, respectively, and dose 2 was 4.84 hours and 1 hour, respectively.

[0635] Table 4. Descriptive Statistics of Serum Pharmacokinetic Parameters of SelK2

[0636] PD assay

[0637] The PD assay was utilized to measure the relative inhibition of Psel-lg binding to PSGL-1 (GSP6) by SelK2 in serum samples using Surface Plasmon Response technology.

[0638] A streptavidin coated gold sensor chip was first loaded with biotinylated GSP6 and diluted serum samples were injected allowing for active SelK2 to bind GSP6. Last, Psel-lg is injected to bind any free GSP6 and the signal was assessed by the change in response units (RU) from before the Psel-lg injection to 25 seconds following injection and non-specificsignal was subtracted using a control, in -line reference channel consisting of streptavidin without GSP6. The relative inhibition is determined by comparing the Psel-lg signal of a serum sample to the Psel-lg signal of the negative control (the specific "pre-dose").

[0639] As shown in FIG. 9, Psel-lg is blocked concurrently in accordance with the increase observed in SelK2 serum concentration. When the mean serum concentration is 35-50 μg / mL, the percentage inhibition of Psel-lg is above 80%; in this assay, 80% or above is considered to be completely blocked. Notably, a 58.6% inhibition of Psel-lg is still achieved with the lowest serum concentration of 27.3 μg / ml at day 57 (+ / - 3 days) following dose 2.

[0640] Example 6. Study to Assess the Safety and Efficacy of SelK2 on Airway Responses Following Allergen Challenge in Subjects with Asthma and COPP (ClinicalTrials.gov Identifier: NCT04540042)

[0641] In a recent two part, randomised, double-blind, placebo-controlled, phase II parallel group study SelK2 was administered to humans. The main purpose of Part 1 of this study was to examine how safe and effective two doses of SelK2 was on participants with mild asthma. Lung function and inflammatory cell numbers were measured in response to the administration of an allergen into the lungs in the presence or absence of SelK2. Part2 of this study examined how safe and effective one doseof SelK2 is on participants with chronicobstructive pulmonary disease (COPD). Lung function and inflammatory cell numbers were measured in COPD patients in the presence or absence of SelK2.

[0642] Key Inclusion Criteria for Part 1 :

[0643] • Males or females, 18-65 years of age (inclusive);

[0644] • Body Mass Index (BMI) ≥ 18.0 and ≤ 35.0 kg / m2.

[0645] • Documented physician-diagnosed asthma for ≥ 4 months prior to screening.

[0646] • Pre-bronchodilator FEV1 ≥ 70% predicted at screening.

[0647] • Documented allergy to at least one common allergen as confirmed by the skin prick test.

[0648] • Dual responder to inhaled bronchial allergen challenges as manifested by positive allergen-induced early (EAR) and late airway broncho constriction (LAR) at screening.

[0649] Key Exclusion Criteria for Part 1 :

[0650] • Lung disease other than stable, mild asthma; e.g., worsening of asthma that requires a change in asthma therapy in the past 4 weeks or is deemed clinically significant by the investigator. • A diagnosed current or recent (within previous 8 weeks of screening, or prior to randomisation) bacterial, protozoal, viral or parasitic infection; is suspected of or is at high risk of having a parasitic infection, or has a history of more than one episode of herpes zoster infection.

[0651] • Has a history of life-threatening asthma, defined as an asthma episode that required intubation and / or was associated with hypercapnea, respiratory arrest and / or hypoxic seizures.

[0652] • Has been hospitalised or has attended the emergency room for asthma in the 12 months prior to screening, or prior to randomization.

[0653] • A history of tuberculosis (latent or active) or systemic fungal diseases.

[0654] Key Inclusion Criteria for Part 2:

[0655] • Male or female, 40 to 75 years of age, inclusive, at the time of informed consent.

[0656] • Confirmed diagnosis by a physician of COPD with symptoms compatible with COPD for at least 1 year prior to screening.

[0657] • BMI ≥ 18.0 and ≤ 35.0 kg / m2 at screening.

[0658] • Able to tolerate sputum induction and produce an adequate sputum sample with a neutrophil differential count > 55% at screening.

[0659] • Post-bronchodilator FEV1 ≥ 30% and ≤ 80% of the predicted normal, and post- bronchodilator FEV1 / FVC < 0.7 at the time of Screening.

[0660] • Current or former tobacco smoker who has a smoking history of at least 10 pack years (T en pack- years are defined as 20 cigarettes a day for 10 years, or 10 cigarettes a day for 20 years).

[0661] • Has a negative result in the blood test for tuberculosis (TB) at screening.

[0662] Key Exclusion Criteria for Part 2:

[0663] • COPD exacerbation requiring oral steroids and / or antibiotics, within the 8 weeks prior to screening or prior to randomisation.

[0664] • A positive sputum culture at Screening indicating ongoing infection.

[0665] • Other respiratory disorders: Subjects with a current diagnosis of asthma, active tuberculosis, lung cancer, bronchiectasis, sarcoidosis, lung fibrosis, interstitial lung diseases, known alpha-1 antitrypsin deficiency or other active pulmonary diseases other than COPD.

[0666] • A history of life-threatening COPD including intensive care unit admission and / or requiring intubation within the last 5 years.

[0667] • A history of > 1 hospitalisation for COPD in the previous 1 year prior to screening. • Previous lung resection, lung reduction surgery or lung transplantation.

[0668] • Requires supplemental oxygen, even on an occasional basis.

[0669] • Any infection requiring hospitalisation or intravenous antibiotics within 6 months prior to Screening or prior to randomisation.

[0670] • A diagnosed current or recent (within previous 8 weeks of screening, or prior to randomisation) bacterial, protozoal, viral or parasitic infection; is suspected of or is at high risk of having a parasitic infection, or has a history of more than one episode of herpes zoster infection.

[0671] • Active participation in a pulmonary rehabilitation program.

[0672] • A history of tuberculosis (latent or active) or systemic fungal diseases.

[0673] The bronchial allergen challenge was performed, as per standard procedures using the method described by Taylor and colleagues (Taylor, Harris, and O’Connor, 2000; Comparison of incremental and bolus dose inhaled allergen challenge in asthmatic patients; Clinical and Experimental Allergy, 30: 56-63), to confirm the presence of an early and late phase response at the screening visit. The cumulative dose of allergen required to achieve a successful response at Screening phase was administered as a bolus during the treatment phase. Subjects that experienced a significant symptomatic fall in FEV1 during the EAR and / or LAR phases at screening (beyond the expected values of 20% and 15% respectively) were only randomised at the discretion of the Investigator. Alternatively, the subject may have repeated the bronchial allergen challenge at screening to achieve a lower cumulative dose of the same allergen or used a different allergen if deemed safe to do so by the Investigator. In the event that the subject met both the EAR and the LAR and a repeat allergen challenge was performed, the repeat challenge was at least 21 days later.

[0674] Time-points for FEV1 relative to the allergen challenge are listed below:

[0675] Screening: Pre-diluent, post-diluent, +5 minutes (multiple depending on number of allergen titrations), +10 minutes (multiple depending on number of allergen titrations), +15 minutes (multiple depending on number of allergen titrations), +20 minutes (multiple depending on number of allergen titrations), +30 minutes (multiple depending on number of allergen titrations), +45 minutes, 1 , 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8 and 24 hours.

[0676] Treatment Phase: Pre-diluent, post-diluent, +5, 10, 15, 20, 30, 45 minutes, 1 , 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8 and 24 hours. Failure to achieve both an EAR and LAR during the allergen challenge conducted at Screening indicated that the subject is non-responsive to the allergen used in the challenge. As asthmatic subjects tend to be atopic to more than one type of air borne allergen, the allergen challenge may be repeated with a different allergen from that used in the initial challenge, if deemed safe by the Investigator. The repeated allergen challenge test, if needed, was performed at least 7 days later.

[0677] Spirometry (FEV1 , FVC, FEV1 % predicted, FVC % predicted, FEV1 / FVC (reported as %), FEF25%-75%, FEF25%-75% % predicted) was measured using a spirometer that meets the American Thoracic Society (ATS) / European Respiratory Society (ERS) recommendations. Spirometry was performed in accordance with ATS / ERS 2019 criteria. Predicted normal values used are based on the Global Lung Function Initiative predicted values (GLI) (2012) (Quanjer, eta / ., 2012; Multi-ethnicreference valuesforspirometryforthe 3-95-year age range: the global lung function 2012 equations; Eur Respir J', 40: 1324-1343).

[0678] Table 5. Phase II Part 1 Study Design.

[0679] As outlined in T able 5, the Phase 11 part 1 study involved 42 days of screening prior to SelK2 or placebo treatment intravenously administered on days 1 and 22. This was followed by a subsequent allergen challenge on day 36. As outlined in Tables 6 and 7, the study was composed of 9 patients who received the SelK2 antibody treatment and 15 patients who received the placebo treatment. The primary and secondary endpoints were assessed on day 36.

[0680] As depicted in Tables 7 and 8, Subject Disposition and Demographics was fairly evenly distributed between SelK2- and Placebo-treated Patients.

[0681] Table 6. Subject Disposition (Part 1)

[0682] Abbreviations: n = the number of subjects meeting the criterion; StdDev = standard deviation. Study duration has been derived as the number of days between Study Day 1 and the date of study completion or the date of early study withdrawal. Table 7. Demographic Characteristics (Part 1)

[0683] Abbreviations: BMI = body mass index; max = maximum; min = minimum; StdDev = standard deviation. a. Age is calculated in years from the date of first study treatment. b. BMI is calculated as weight (kg)Zheight (m2).

[0684] FIG. 10 shows the allergen challenge response curve (EAR and LAR) of placebo patients during screening and during treatment. If the allergen challenge model is well controlled, the two curves should lay on top of one another as demonstrated in the figure. T able 8 and FIG. 11 show that treatment with SelK2 antibody in comparison to the placebo control reduced the area under the curve (AUC) of percentage fall in FEV1 between 3 and 8 hours post allergen challenge resulting in a mean percentage improvement in lung function of 45.7%. Moreover, as shown in Table 9 and FIG. 1 1 , treatment with SelK2 antibody in comparison to the placebo control reduced the mean maximum percent fall between 3 and 8 hours post allergen challenge resulting in a mean improvement in lung function of 52.4%.

[0685] Table 8. Statistics of AUC of % fall in FEV1 between 3 and 8 hours

[0686] Table 9. Statistics of Maximum % fall in FEV1 between 3 and 8 hours

[0687] As shown in FIGS. 12 and 14, asthmatic patients who received the SelK2 treatment demonstrated a significant reduction in absolute differential eosinophil count and percentage differential eosinophil count respectively at 8 hours and 24 hours post-challenge in comparison to patients receiving the placebo treatment. FIG. 12 demonstrates that the mean absolute differential eosinophil count (10E6 / g) was reduced from 1 .15 to 0.44 at 8 hours post-challenge and from 1.36 to 0.47 at 24 hours post-challenge, in response to SelK2 treatment.

[0688] FIG. 14 demonstrates that the percentage differential eosinophil count was reduced from 17.2% to 8.6% at 8 hours post-challenge and from 18.7% to 8.3% at 24 hours post- challenge in response to SelK2 treatment. In contrast, FIGS. 13 and 15 show that the reduction in eosinophils was not observed at 8 hours and 24 hours post-challenge in the blood of asthmatic patients. Thus, the reduction in eosinophils in the lungs with SelK2 treatment is not due to eosinophil depletion in the bloodstream that is characteristic of other approved therapies. Rather, the reduction of eosinophils in the lungs with SelK2 treatment is due to a decrease in extravasation of these cells across the endothelium and into the lungs.

[0689] This is supported by FIG. 16, which demonstrates that COPD patients who received the SelK2 treatment exhibited a significant reduction in absolute differential eosinophil count in sputum between days 8 and 28 of the study in comparison to patients receiving the placebo treatment. FIG. 16 demonstrates that at day 15, the mean absolute differential eosinophil count (10E6 / g) was significantly reduced from 0.198 to 0.045; at day 22, the mean absolute differential eosinophil count (10E6 / g) was significantly reduced from 0.229 to 0.016; and at day 28, the mean absolute differential eosinophil count (10E6 / g) was reduced from 0.187 to 0.048.

[0690] Moreover, FIG. 17 shows that the reduction in eosinophils was not observed in the blood of COPD patients who received the SelK2 treatment.

[0691] Finally, FIG. 18 demonstrates howCOPD patients who received the SelK2 treatment exhibited a significant reduction in absolute differential epithelial count in sputum following day 22 of the study in comparison to patients receiving the placebo treatment. At day 22, the mean absolute differential epithelial count (10E6 / g) was significantly reduced from 0.350 to 0.038; at day 29, the mean absolute differential epithelial count (10E6 / g) was significantly reduced from 0.286 to 0.128; and at day 43, the mean absolute differential epithelial count (10E6 / g) was significantly reduced from 0.332 to 0.128.

[0692] Thus, this Phase II study confirms the role of SelK2 antibody in reducing eosinophil migration into the lungs resulting in an improvement in lung function post-allergen challenge in asthmatic patients. Importantly, the levels of eosinophils in the blood did not change, indicating that systemic immune suppression of these cells did not occur. Example 7. Comparative analysis of airway response following an allergen challenge in patients with stable allergic asthma treated with SelK2 and Tezepelumab antibodies

[0693] FIG. 19 is a comparative analysis demonstrating the percentage fall in FEV1 in patients with stable allergic asthma treated with the SelK2 antibody or the recently approved T ezepelumab antibody. Patients in both groups were subjected to an allergen challenge on day 36 in the case of SelK2 and on day 42 in the case of T ezepelumab. The T ezepelumab graph is taken from a previously published study that demonstrates the percentage FEV1 response in 16 patients with mild asthmatic patients treated with T ezepelumab antibody or a placebo control and subjected to an allergen challenge (Gauvreau, et al., 2014, Effects of an Anti-TSLP Antibody on Allergen-Induced Asthmatic Responses; The New England Journal of Medicine; 370: 2102-21 10).

[0694] As shown in Table 12 and FIG. 19, treatment with Tezepelumab antibody resulted in a 34% improvement in the maximum percentage fall in FEV1, a 33% improvement in FEV1AUC, and a 4% improvement in average LAR percent fall. By contrast, SelK2 treatment resulted in a 55% improvement in the maximum percentage fall in FEV1, a 51 % improvement in FEV1AUC, and a 15% improvement in average LAR percent all. All three parameters of lung function following allergen challenge were superior with SelK2 treatment versus Tezepelumab treatment. In addition, both drugs resulted in a marked decrease of eosinophils in the lungs.

[0695] Table 10. SelK2 antibody and Tezepelumab antibody airway response following an allergen challenge

[0696] Example s. Immunogenicity analysis

[0697] Immune responses to the antibodies of the invention were investigated in recipients who had received SelK2 or placebo as part of the clinical trial.

[0698] Sera from each patient collected at the specified time points was tested in triplicate using an AlphaLISA format. Diluted sera was mixed with Donor and Acceptor beads previously coated with SelK2 Antibody. If anti-SelK2 antibodies are present in the sera, the anti-antibodies crosslink the two bead types, which causes an increase in fluorescence. Combined test results from pre-dose sera were used to establish the Positive / Negative cutoff. Triplicate measurements of pre-dose sera (collected prior to the initial loading dose) was used to calculate the mean and standard deviation of the measurement. The number of data points used to calculate the standard deviation was three times the number of pre-dose specimens collected. The 95% Positive / Negative cutoff was determined using the following formula: 95% Upper Cutoff = Average Signal + (1 .96 X Standard Deviation of all Signals)

[0699] Results

[0700] The results of this analysis are set out in T able 11. As can be seen, incidences of immune responses were 27.8% of SelK2-treated subjects and 16.7% of placebo-treated subjects. Further, immune responses were of low levels, some of which did not repeat at later time points, and no neutralization of the activity of the antibody of the invention was observed. Table 11. Immune responses to the SelK2 in comparison to placebo control.

[0701] *Responses were of low levels, some of which did not repeat at later time points, and no neutralization of the activity of the antibody of the invention was observed.

[0702] Example 9. Study to evaluate the effect of SelK2 on absolute cell counts for immune cells in sputum of COPP patients.

[0703] In a recent randomised, double-blind, placebo-controlled, parallel group study, the effect of SelK2 on sputum inflammatory cells in subjects with a diagnosis of COPD was investigated in comparison to placebo control. The main purpose of this study was to test the hypothesis that SelK2 will inhibit or disrupt extravasation of inflammatory cells into the airways of the lung in subjects with COPD and to evaluate the safety and efficacy of intravenously administered SelK2 compared with placebo. It was planned to recruit approximately 24 subjects with a diagnosis of COPD who had a sputum neutrophil count of >55% at Screening. Subjects were randomized in a 2:1 ratio to receive either 7.5 mg / kg SelK2 or placebo, respectively.

[0704] All subjects received a single dose of study drug on Day 1 . The appropriate unit dose of SelK2 was prepared (by unblinded pharmacy staff or designee) as a SelK2 saline admixture to deliver the target dose to a subject administered a 100 mL infusion.

[0705] Table 12. Change from baseline in absolute numbers of eosinophils and neutrophils in sputumand in total numberof cells pergram of sputum on Day 22 (21 daysafterdosing) shown in FIGs. 20, 21 and 22 and in Table 12, statistically significant reductions from baseline

[0706] 1p-value was calculated based on the least squares mean. were seen at Day 22 in absolute eosinophil and neutrophil cell counts and in total cells per gram of sputum in SelK2-treated subjects compared to placebo-treated subjects (p = 0.0181 , 0.0413 and 0.0257, respectively, based on adjusted least squares (LS) means).

[0707] Looking specifically at the reduction in absolute numbers of neutrophils in the SelK2 treatment group, cell counts went from a mean of 4.1 + / - 3.1 x 106 / g at baseline to 1 .7 + / - 1 ,4 x 106 / g at Day 22, while cell counts in the placebo group were unchanged at a mean of 5.8 + / - 8.4 x 106 / g at baseline and 5.8 + / - 7.5 x 106 / g at Day 22 (p = 0.0413, based on adjusted LS means).

[0708] T urning to the reduction in absolute numbers of eosinophils in the SelK2 treatment group, cell counts went from a mean of 0.10 + / - 0.09 x 106 / g at baseline to 0.02 + / - 0.02 x 106 / g at Day 22, while eosinophil counts in the placebo group increased from a mean of 0.17 + / - 0.22 x 106 / g at baseline to 0.23 + / - 0.35 x 106 / g at Day 22 (p = 0.0181 , based on adjusted LS means).

[0709] Similarly, total number of cells per gram of sputum in the SelK2 treatment group went from a mean of 4.8 + / - 0.8 x 106 / g at baseline to 2.0 + / - 1.5 x 106 / g at Day 22, while counts in the placebo group went from a mean of 7.1 + / - 3.5 x 106 / g at baseline to 7.4 + / - 9.7 x 106 / g at Day 22 (p = 0.0257, based on adjusted LS means).

[0710] Neutrophils and eosinophils are both implicated in the pathogenesis of COPD, and so the ability to achieve targeted reduction of these cell types in the lungs with a single dose of SelK2 is encouraging. The chronic nature of COPD means that it is unexpectedly positive that a reduction of this sort is able to be achieved within the relatively short treatment period assessed. Prolonged treatment with SelK2 may be hypothesised to yield further beneficial effects.

[0711] While numbers of neutrophil and eosinophils present in the sputum of COPD patients were reduced, corresponding changes in numbers of neutrophils and eosinophils were not observed in the blood of SelK2 treatment group subjects. Without being bound by theory, this indicates that the reduction in neutrophils and eosinophils may result from the inhibition of extravasation of these cells into the lungs and not a reduction in the number of these cells in the circulation. Notably, previously published therapies have relied upon the depletion of these inflammatory cells in the bloodstream in order to achieve a desired depletion in the lungs. Since SelK2 specifically inhibits extravasation of these cells into the lungs, while leaving the cell counts in the blood intact it may be expected that SelK2 treatment will not adversely impact other beneficial functions of these cells in the blood. Example 10: Generationofa comparatorproduct-formulationdevelopmentto develop a high concentration stable formulation for SelK2.

[0712] This Example describes a study undertaken by the inventors to design a high concentration formulation suitablefordeliveryofan antibody exemplified by SelK2 (the anti-PSGL-1 antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2) by following a standard screening procedure in a rational design approach for formulation development. The resulting formulation is not a pharmaceutical composition of the invention, but represent a comparator product that a skilled person, seeking to develop a high concentration stable formulation for SelK2, would generate by follow typical approaches to formulation design.

[0713] The aim of the study was to develop a high concentration stable formulation for SelK2 that would have characteristics suitable for subcutaneous administration. With that in mind, the the following product profile was targeted:

[0714] • SelK2 antibody concentration of ≥150mg / mL

[0715] • pH range of 5-7

[0716] • Osmolality of 290 + / - 10 mOsmo / kg

[0717] • Viscosity of <20cP

[0718] At the start of formulation development, buffers and pH were evaluated for SelK2 formulation development. Dialysis was performed on SelK2 at 10 mg / mL with 13 different buffers at different pH levels. A 3-12 mL dialysis cartridge was used to put 10 mL of SelK2 in a beaker with each of the 13 buffers, including the control (CF). The buffer in each beakerwas replaced twice, once after overnight dialysis. The formulations were removed from the cartridges and the concentration was measured via A280. The cartridge containing water showed visible particulates. The formulations were then filtered by 0.22 μm PES filter and concentration was measured again via A280. Table 40 shows the amount of each sample that was retrieved from the cartridge along with the concentration before filtration, showing total protein recovery out of 100 mg. All formulations were slightly yellow in colour with no visible particulates. One ml of formulations was dispensed into 3 ml vial and stored at 50°C for 7 and 14 days. Table 13: Dialysis Recovery

[0719] Determination of Tm and Tagg

[0720] The formulations at different pH levels were temperature ramped from 20°C to 80°C at a rate of 2°C / minute. Fluorescence data was collected at 30-second intervals. RALS and IF data were collected simultaneously using the same sample. The transition temperatures Tagg and Tm were ascertained from their respective inflection points on the curve.

[0721] Table 14: Tm and Tagg: Buffer / pH screen

[0722] Based on I F, there are two thermal transition T m for all formulations except forthe CF and the formulation in water (minor transition). Amongst the formulations containing buffers, the second transition offormulationscontaining acetate (pH 5.0) suggests unfolding at slightly a higher temperature of 79°C. Based on visual observations afterthe temperature ramping, the formulations containing acetate 5.0 and histidine 6.0 were slightly less precipitated. Effect of Freeze Thaw

[0723] Freeze-Thaw (FT) of a protein may impact its stability; this impact can be usually mitigated by the proper selection of buffer systems and additives. To understand the impact of FT on SelK2 formulations, the effects offreezing andthawing five times were studied. Aliquotsof the protein in formulation buffer were repeatedly frozen at -80°C and thawed to ambient temperatures for five cycles, then analyzed.

[0724] Osmolality

[0725] The osmolality of Selk2 formulations after freeze thaw were measured via depression in the freezing point. These measurements were made using a Model 3340 freezing-point micro osmometer, equipped with a 20 μL Ease Eject™ Sampler (Part#: 3M0825 and 3M0828) by Advanced Instruments. The units of measure for osmolality are milliosmoles of solute in 1 kg of pure solvent (mOsmo / kg H2O). The instrument was calibrated with 50 mOsmo / kg and 850 mOsmo / kg calibration standards and verified with a 290 mOsmo / kg Clinitrol® control Reference Solution before analysis. Sample testing was conducted in accordance with SOP for refractive index measurement.

[0726] There were no changes in osmolality after freeze thaw. pH

[0727] The pH of the sample after freeze thaw was measured using a Thermo Scientific Orion Star ModelA211 pH meter equipped with a Ross PerpHecT microelectrode (Model 8220BNWP). For buffer solutions preparation, a triode electrode was used to measure the pH (Thermo Scientific, US Gel-filled Ultra Triode Electrodes). The instruments were standardized using pH 4, 7, and 10 buffers traceable to NIST Standard Reference Solutionsfrom Thermo Scientific (P / N: 910410 / 910425 (pH 4); 910710 / 910725 (pH 7); and 911010 / 911025 (pH 10), respectively).

[0728] There were no changes in pH observed after freeze thaw.

[0729] Subvisible particles

[0730] Subvisible particles in the samples were monitored via FlowCam (Fluid Imaging Technologies, Inc.). The instrument was focused on 10 μm polystyrene beads at 3000 / mL NIST standard. Samples were aspirated at 0.08 mL / min through a 100 μm x 1 mm flow cell, and images of the particles were taken with a 10X optics system. Particles per mL were calculated using Visual Spreadsheet software. In between samples, the flow cell was rinsed with hot deionized water, 70% 2-propanol, or placebo. Table 15: FlowCam T0 results

[0731] All formulations showed an upward trend in the number of particulates after five cycles of freeze thaw except for acetate pH 5. The formulations containing histidine had relatively lower numbers of particulates compared to the other formulations.

[0732] Particle size analysis

[0733] The particle size analysis was carried out using Malvern Zetasizer was followed. The parameters used for DLS measurements are as shown in Table 16.

[0734] Table 16: The parameters used for DLS measurements Table 17: Dynamic Light Scattering after freeze thaw cycles

[0735] The z-average size of the formulations highlighted in Table 17 (Acetate, citrate and succinate at pH 5.5) remained relatively unchanged; the rest of the formulations showed changes in size, as monitored by DLS (Table 17).

[0736] Size-Exclusion Chromatography HPLC (SEC)

[0737] After FT, 100 μL of sample was removed and then centrifuged. Fifty μL was drawn from the top of the vial and diluted to 1 mg / ml and analysed for HPLC analysis. Agilent 1100 was used for SEC-HPLC. It was specified <10%RSD for intermediate precision and >0.98 for linearity coefficient.

[0738] Table 18: SelK2 Size-Exclusion Chromatography Method Parameters Table 19: Effect of FT: SEC-HPLC

[0739] All formulations performed similarly afterfreeze thaw exceptforthe formulation in water, which had the lowest recovery. There is a slight increase in %HM W peak after FT for all formulations.

[0740] Effect of Simulated Shear Stress

[0741] For simulated shear stress, experiments were performed in 1.5 mL Reacti-Vials™ (Thermo Scientific) with matching triangular stir bars and stirred at 900 rpm for 24 hours.

[0742] B...

Claims

CLAIMS1 . A pharmaceutical composition comprising:• an antibody at a concentration of approximately 210 mg / mL;• 10mM acetate;• 110mM sorbitol;• 0.1 % poloxamer 188;• 50mM proline and;• 110mM mannitol;• wherein the composition has a pH of 5.3; and• the antibody comprises or consists of an antibody heavy chain of SEQ ID NO: 1 and an antibody light chain of SEQ ID NO: 2, or a fragment or variant of such an antibody sharing at least 90% sequence identity with an antibody that consists of SEQ ID NO: 1 and SEQ ID NO:2.

2. A pharmaceutical composition comprising:• an antibody at a concentration of at least 150 mg / mL;• 9m M to 11 mM acetate;• 50mM to 220mM sorbitol;• 0.05% to 0.15% poloxamer 188;• 45mM to 55mM proline; and• 100mM to 220mM mM mannitol;• wherein the composition has a pH of approximately 5.3; and• the antibody comprises or consists of an antibody heavy chain of SEQ ID NO: 1 and an antibody light chain of SEQ ID NO: 2, ora fragment orvariant of such an antibody sharing at least 90% sequence identity with an antibody that consists of SEQ ID NO: 1 and SEQ ID NO:2.

3. The pharmaceutical composition accordingto claim 2, comprising acetate at a concentration of approximately 10 mM.

4. The pharmaceutical composition according to claim 2 or claim 3, comprising sorbitol at a concentration of approximately 75 to 150 mM.

5. The pharmaceutical composition according to any of claims 2 to 4, comprising poloxamer at approximately 0.1 %.

6. The pharmaceutical composition according to any of claims 2 to 5, comprising proline at a concentration of approximately 50 mM.

7. The pharmaceutical composition according to any of claims 2 to 6, comprising mannitol at a concentration of approximately 100 mM to 150 mM.

8. The pharmaceutical composition of any preceding claim, comprising acetate provided as sodium acetate.

9. The pharmaceutical composition of any preceding claim, wherein the osmolality of the composition is approximately 290 mOsm / kg.

10. The pharmaceutical composition of any preceding claim, wherein the absolute viscosity of the composition is 20 cP or less.

11. The pharmaceutical composition of any preceding claim, with an injection glide force of less than or equal to 25 N for a 25G needle.

12. The pharmaceutical composition of any preceding claim, comprising an anti-PSGL-1 antibody.

13. The pharmaceutical composition of any claim 12, wherein the antibody has an affinity for PSGL-1 of below 5 nM.

14. The pharmaceutical composition of claim 13, wherein the antibody has an affinity for PSGL- 1 of approximately 4.29 nM.

15. The pharmaceutical composition of any of claims 12 to 14, wherein the antibody is able to inhibit and disrupt binding of PSGL-1 to at least one ligand.

16. The pharmaceutical composition of claim 15, wherein the antibody is able to inhibit and disrupt binding of PSGL-1 to at least one selectin.

17. The pharmaceutical composition of any preceding claim 15 or claim 16, whereinthe antibody is able to inhibit and disrupt binding of PSGL-1 to at least one chemokine.

18. The pharmaceutical composition of any of claims 15 to 17, wherein the antibody is able to inhibit and disrupt binding of PSGL-1 to VISTA.

19. The pharmaceutical composition of any preceding claim, wherein the antibody, orfragment or variant, comprises six CDRs, each of which shares at least at least 90% sequence identity with the sequences of the corresponding CDRs set out in SEQ ID NO: 1 and SEQ ID NO: 2.

20. The pharmaceutical composition of any of any preceding claim, wherein the antibody, or fragment or variant, comprises each of the three CDRs from the antibody heavy chain sequence set out in SEQ ID NO: 1 and each of the three CDRs from the antibody light chain sequence set out in SEQ ID NO: 2.21 . The pharmaceutical composition of any of any preceding claim, wherein the antibody, or fragment or variant, comprises a heavy chain variable region set out in SEQ ID NO: 3.

22. The pharmaceutical composition of any preceding claim, wherein the antibody, orfragment or variant, comprises a light chain variable region set out in SEQ ID NO: 4.

23. The pharmaceutical composition of any preceding claim, wherein the antibody comprises an antibody comprising SEQ ID NO: 1 and SEQ ID NO: 2.

24. The pharmaceutical composition of claim 23, wherein the antibody consists of an antibody comprising SEQ ID NO: 1 and SEQ ID NO: 2.

25. The pharmaceutical composition of any preceding claim, wherein the antibody comprises an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2.

26. The pharmaceutical composition of claim 25, wherein the antibody consists of an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2.

27. The pharmaceutical composition of any of claims 1 to 18, wherein the antibody comprises an antibody, fragment, orvariantsharing at Ieast95%sequence identity with an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2.

28. The pharmaceutical composition of claim 27, wherein the antibody consists of an antibody, fragment, or variant sharing at least 95% sequence identity with an antibody consisting of SEQ ID NO: 1 and SEQ ID NO: 2.

29. The pharmaceutical composition of any preceding claim, for use as a medicament.

30. A method of preventing or treating a disease or condition in a subject in need thereof, the method comprising providing to the subject a therapeutically effective amount of a pharmaceutical composition of any of claims 1 to 29.31 . The method of claim 30, for the prevention or treatment of an inflammatory condition or disease.

32. The method of claim 31 , for the prevention or treatment of an inflammatory condition or disease selected from the group consisting of: asthma; chronic obstructive pulmonary disease; allergic reactions; inflammatory bowel disease (e.g., Crohn's disease, ulcerative colitis, enteritis); arthritis (e.g., rheumatoid arthritis, osteoarthritis, psoriatic arthritis); graftrejection; graft versus host disease; psoriasis; dermatitis; nephritis; lupus erythematosus; scleroderma; rhinitis; anaphylaxis; diabetes; multiple sclerosis; atherosclerosis; and thyroiditis.

33. The method of any of claims 30 to 32, for the prevention or treatment of an eosinophilic condition or disease selected from the group consisting of: eosinophilic asthma; allergies; eosinophilic oesophagitis; eosinophilic dermatitis; acute myelogenous leukemia (AML), ascariasis; atopic dermatitis (eczema); bullous pemphigoid; cancer (such as Hodgkin lymphoma, leukemia, and certain myeloproliferative neoplasms); Churg-Strauss syndrome; drug allergy; eosinophilic cardiomyopathy; eosinophilic cellulitis (Wells’ syndrome); eosinophilic colitis; eosinophilic enteritis;; eosinophilic fasciitis; eosinophilic gastrointestinal diseases; eosinophilic granulomatosis with polyangiitis (EGPA); eosinophilic leukemia; eosinophilic myocarditis; hay fever (allergic rhinitis); Hodgkin's lymphoma (Hodgkin's disease); hypereosinophilic syndromes; idiopathic hypereosinophilic syndrome (HES); lgG4-Related Disease; inflammatory bowel disease (Crohn's disease, ulcerative colitis); lymphaticfilariasis; neuromyelitisoptica(NMO); ovarian cancer; parasitic infection; primary biliary cirrhosis; primary immunodeficiency; or trichinosis.

34. The method of claim 30 for the prevention or treatment of asthma.

35. The method of claim 30 for the prevention or treatment of COPD.

36. The method of claim 30 for the treatment of cancer.

37. The method of claim 36 for the treatment of cancer selected from the group comprising melanoma, sarcoma, lymphoma, central nervous system (CNS) cancer, CNS germ cell tumour, lung cancer, leukaemia, multiple myeloma, renal cancer, glioma, medulloblastoma, ovarian cancer, breast cancer, prostate cancer, bladder cancer, pancreatic cancer, gastric cancer, head and neck cancer, mesothelioma, non-melanoma skin cancer, and colorectal cancer; optionally wherein (i) thesarcoma is a fibrosarcoma; (ii) the lymphoma is follicular lymphoma, Hodgkin's lymphoma, or non- Hodgkin's lymphoma; (iii) the lung cancer is non-small cell lung cancer; (iv) the renal cancer is renal cell carcinoma; (v) the ovarian cancer is epithelial ovarian cancer; (vi) the breast cancer is triple negative breast cancer; (vii) the non-melanoma skin cancer is merkel cell carcinoma; and / or (viii) the colorectal cancer is colon adenocarcinoma.

38. The method of claim 36 or claim 37, for the treatment of cancer selected from the group comprising undifferentiated carcinoma; giant and spindle cell carcinoma; small cell carcinoma; papillary carcinoma; squamous cell carcinoma; lymphoepithelial carcinoma; basal cell carcinoma; pilomatrix carcinoma; transitional cell carcinoma; papillary transitional cell carcinoma; adenocarcinoma; gastrinoma; cholangiocarcinoma; hepatocellular carcinoma; combined hepatocellular carcinoma and cholangiocarcinoma; trabecular adenocarcinoma; adenoid cystic carcinoma; adenocarcinoma in adenomatous polyp; familial polyposis coli adenocarcinoma; solid carcinoma; carcinoid tumour; branchiolo-alveolar adenocarcinoma; papillary adenocarcinoma; chromophobe carcinoma; acidophil carcinoma; oxyphilic adenocarcinoma; basophil carcinoma; clear cell adenocarcinoma; granular cell carcinoma; follicular adenocarcinoma; papillary and follicular adenocarcinoma; non-encapsulating sclerosing carcinoma; adrenal cortical carcinoma; endometroid carcinoma; skin appendage carcinoma; apocrine adenocarcinoma; sebaceous adenocarcinoma; ceruminous; adenocarcinoma; mucoepidermoid carcinoma; cystadenocarcinoma; papillary cystadenocarcinoma; papillary serous cystadenocarcinoma; mucinous cystadenocarcinoma; mucinous adenocarcinoma; signet ring cell carcinoma; infiltrating duct carcinoma; medullary carcinoma; lobular carcinoma; inflammatory carcinoma; mammary Paget's disease; acinar cell carcinoma; adenosquamous carcinoma; thymoma; ovarian stromal tumour; thecoma; granulosa cell tumour; roblastoma; Sertoli cell carcinoma; Leydig cell tumour; lipid cell tumour; paraganglioma; extra-mammary paraganglioma; pheochromocytoma; glomangiosarcoma; amelanotic melanoma; superficial spreading melanoma; epithelioid cell melanoma; blue nevus; fibrous histiocytoma; myxosarcoma; liposarcoma; leiomyosarcoma; rhabdomyosarcoma; embryonal rhabdomyosarcoma; alveolar rhabdomyosarcoma; stromal sarcoma; mullerian mixed tumour; nephroblastoma; hepatoblastoma; carcinosarcoma; mesenchymoma; brenner tumour; phyllodes tumour; synovial sarcoma; dysgerminoma; embryonal carcinoma; teratoma; struma ovarii; choriocarcinoma; mesonephroma; hemangiosarcoma; hemangioendothelioma; kaposi's sarcoma; hemangiopericytoma; lymphangiosarcoma; osteosarcoma; juxtacortical osteosarcoma; chondrosarcoma; chondroblastoma; mesenchymal chondrosarcoma; giant cell tumour of bone; Ewing's sarcoma; odontogenic tumour; ameloblastic odontosarcoma; ameloblastoma; ameloblastic fibrosarcoma; pinealoma; chordoma; ependymoma; astrocytoma; protoplasmic astrocytoma; fibrillary astrocytoma; astroblastoma; glioblastoma; oligodendroglioma; oligodendroblastoma; primitive neuroectodermal; cerebellar sarcoma; ganglioneuroblastoma; neuroblastoma; retinoblastoma; olfactory neurogenic tumour; meningioma;neurofibrosarcoma; neurilemmoma; granular cell tumour; paragranuloma; small lymphocytic lymphoma; diffuse large cell lymphoma; mycosis fungoides; histiocytosis; mast cell sarcoma; immunoproliferative small intestinal disease; lymphoid leukaemia; plasma cell leukaemia; erythroleukemia; lymphosarcoma cell leukaemia; myeloid leukaemia; basophilic leukaemia; eosinophilic leukaemia; monocytic leukaemia; mast cell leukaemia; megakaryoblastic leukaemia; myeloid sarcoma; and hairy cell leukaemia.

39. The method of any of claims 36 to 38 for the treatment of a primary tumour.

40. The method of any of claims 36 to 39 for the treatment of cancer resistant to an immune checkpoint inhibitor and / or wherein the subject has previously been administered an immune checkpoint inhibitor.41 . The method of any of claims 30 to 40, wherein the pharmaceutical composition is provided in combination with an immune checkpoint inhibitor.

42. The method of claim 41 , wherein the pharmaceutical composition is administered sequentially with the immune checkpoint inhibitor.

43. The method of claim 41 , wherein the pharmaceutical composition is administered simultaneously with the immune checkpoint inhibitor.

44. The method of any of claims 41 to 43, wherein the immune checkpoint inhibitor comprises an inhibitor of CTLA-4, PD-1 , PD-L1 , PD-L2, TIM-3, LAG3, TIGIT, CD160, B7H3, B7H4, B7H6, BTLA, VISTA, LAIR1 , and / or LAIR2.

45. The method of claim 44, wherein the immune checkpoint inhibitor is an antibody.

46. The method of claim 30 for the treatment of an infection.

47. The method of claim 46, wherein the infection is an acute infection.

48. The method of claim 47, wherein the acute infection is infection with influenza or a coronavirus (such as SARS-CoV-2).

49. The method of claim 46, wherein the infection is a chronic infection.

50. The method of claim 49, wherein the chronic infection is selected from infection with human immunodeficiency virus (HIV), Hepatitis B, Hepatitis C, herpes simplex virus, varicella zoster virus, cytomegalovirus, Human T-lymphotropic virus 1 (HTLV-1 ), mumps virus, rubella virus, measles virus, poliovirus, Epstein-Barr virus, SARS-CoV-2 (e.g., “long COVID”), Mycobacterium tuberculosis, Helicobacter pylori, Salmonella Typhi, Treponema pallidum, Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus (e.g., MRSA), Hemophilus influenza, Mycobacterium leprae, Candida (e.g., Candida albicans), Malassezia, Sporothrix, Histoplasma, Coccidioides, Blastomyces, Aspergillus, Pneumocystis (e.g., Pneumocystis jirovecii), Rhizopus, Mucor, Cryptococcus (e.g., Cryptococcus neoformans and Cryptococcus gattii), Plasmodium (e.g., Plasmodium falciparum, Plasmodium malariae, Plasmodium knowlesi, Plasmodium vivax, and Plasmodium ovale), Giardia (e.g., Giardia lamblia), Toxoplasma (e.g., Toxoplasma gondii), Cyclospora, Entamoeba, Isospora, Blastocystis, Strongyloides, Schistosoma (e.g., Schistosoma haematobium, Schistosoma mansoni, and Schistosoma japonicum), Leishmania, Trypanosoma, Opisthorchis viverrini, Clonorchis sinensis, Heligmosomoides polygyrus, Taenia (e.g., Taenia crassicep and Taenia solium), Ascaris lumbricoides, Nercatoramericanus, Trichomonas vaginalis, Entamoeba histolytica, Wuchereria bancrofti, Onchocerca volvulus, Enterobiusvermicularis, and / or Trichuris trichiura.51 . The method of any of claims 46 to 50, wherein the pharmaceutical composition is provided in combination with an anti-infective agent; optionally wherein the anti-infective agent is an anti-viral agent, anti-bacterial agent, anti-fungal agent, anti-parasitic agent, or anti-protozoan agent.

52. The method of claim 51 , wherein the pharmaceutical composition is administered sequentially with the anti-infective agent.

53. The method of claim 52, wherein the pharmaceutical composition is administered simultaneously with the anti-infective agent.

54. The method of any of claims 30 to 53, wherein the pharmaceutical composition is provided in combination with an immunotherapy agent.

55. The method of claim 54, wherein the pharmaceutical composition is administered sequentially with the immunotherapy agent.

56. The method of claim 54, wherein the pharmaceutical composition is administered simultaneously with the immunotherapy agent.

57. A method of treating cancer that is not responsive to checkpoint inhibitor therapy, the method comprising administering to a subject in need thereof a pharmaceutical composition of any of claims 1 to 29, in an amount sufficient to treat the cancer.

58. A method of treating relapsed or refractory cancer, the method comprising administering to a subject in need thereof a pharmaceutical composition of any of claims 1 to 29, in an amount sufficient to treat the cancer.

59. A method of maintaining a reduction in size of a tumour, the method comprising providing a pharmaceutical composition of any of claims 1 to 29 to a subject in need thereof.

60. A method of sensitizing cancer to treatment with an immunotherapy agent, the method comprising providing a pharmaceutical composition of any of claims 1 to 29 to a subject in need thereof.61 . The method of any of claims 30 to 60, wherein the pharmaceutical composition is provided by subcutaneous administration.