Methods of treating or delaying progression or hindering development of a neutrophil-associated condition

Abstract

The present disclosure relates to a method treating or delaying progression or reducing or inhibiting or hindering development of a neutrophil-associated condition in a subject, wherein the subject has had an inadequate clinical response to an anti-tumour necrosis factor (TNF) therapy for the neutrophil-associated condition, the method comprising administering to the subject a compound that inhibits granulocyte colony stimulating factor (G-CSF) signaling.

Description

[0001] METHODS OF TREATING OR DELAYING PROGRESSION OR HINDERING DEVELOPMENT OF A NEUTROPHIL-ASSOCIATED CONDITION

[0002] RELATED APPLICATION DATA

[0003] The present application claims priority from United States provisional application No. 63 / 633,894 entitled “Methods of treating or delaying progression or hindering development of a neutrophil-associated condition” fded 15 April 2024, the entire contents of which is hereby incorporated by reference.

[0004] SEQUENCE LISTING

[0005] The present application is filed together with a Sequence Listing in electronic form. The entire contents of the Sequence Listing are hereby incorporated by reference.

[0006] FIELD

[0007] The present disclosure relates to a method treating or delaying progression or reducing or inhibiting or hindering development of a neutrophil-associated condition in a subject, wherein the subject has had an inadequate clinical response to an anti-TNF therapy for the neutrophil-associated condition, the method comprising administering to the subject a compound that inhibits granulocyte colony stimulating factor (G-CSF) signaling.

[0008] BACKGROUND

[0009] Neutrophils are central to the innate immune response and are one of the first cells to migrate to the site of injury or infection, where they eliminate pathogens or dying cells by phagocytosis, degranulation, and neutrophil extracellular trap formation. In addition, neutrophils promote the inflammatory response by releasing cytokines and chemokines that recruit and activate additional immune cells. However, if not appropriately regulated during the inflammatory response (e.g., delayed apoptosis of neutrophils after elimination of the pathogen), neutrophils can perpetuate inflammation, leading to modification and destruction of target tissues. The perpetuation of inflammation by neutrophils is evident in certain chronic inflammatory diseases where cellular infiltrates are predominantly neutrophils, for example in conditions such as rheumatoid arthritis, Crohn’s disease and skin conditions such as psoriasis and hi dradenitis suppurativa (HS).

[0010] Current first-line treatment options for neutrophil-associated skin conditions include antibiotics, hormonal treatment, clindamycin, corticosteroids, resorcinol and retinoids. If these treatments fail to control the disease and chronic, relapsing, or severe disease persists, surgical excision may be considered. Biologies may also be used for the treatment of moderate to severe neutrophilic skin conditions, for example disease which is unresponsive to systemic antibiotics. For example, anti-tumor necrosis factor (TNF) therapies, such as adalimumab, are approved for the treatment of various skin conditions including moderate to severe HS and psoriasis in the United States and European Union. Adalimumab was effective in approximately 40% to 60% of subjects with HS, with loss of response in approximately 40% to 50% of subjects by Week 36 of treatment.

[0011] Accordingly, based on the foregoing the skilled person will recognise there remains a significant need for improved therapies for treating neutrophil- associated conditions.

[0012] SUMMARY

[0013] In producing the present invention, the inventors identified granulocyte colony stimulating factor (G-CSF) as a potential target for pharmacological intervention of neutrophilic inflammation in patients that have had an inadequate clinical response to an anti-TNF therapy. Through global transcriptomics of skin samples from patients with hidradenitis suppurativa (HS), the inventors found increased expression of genes in the G-CSF gene signature in patients with HS, indicating activation of the G-CSF signalling pathway. Furthermore, the inventors identified the G-CSF gene signature remains elevated in patients who were non-responders to treatment with adalimumab (an antitumor necrosis factor (TNF) alpha (a) therapy) for the HS. Thus, the findings provide the basis for methods of inhibiting G-CSF signaling in patients with neutrophil-associated conditions and who have had an inadequate clinical response to anti-TNF therapy.

[0014] Accordingly, the present disclosure provides a method of treating or delaying progression or reducing or inhibiting or hindering development of a neutrophil- associated condition in a subject, wherein the subject has had an inadequate clinical response to an anti-TNF therapy for the neutrophil-associated condition, the method comprising administering to the subject a compound that inhibits granulocyte colony stimulating factor (G-CSF) signaling.

[0015] The present disclosure also provides a compound that inhibits G-CSF signaling for use in treating or delaying progression or reducing or inhibiting or hindering development of a neutrophil-associated condition in a subject, wherein the subject has had an inadequate clinical response to an anti-TNF therapy for the neutrophil-associated condition.

[0016] The present disclosure further provides use of a compound that inhibits G-CSF signaling in the manufacture of a medicament for treating or delaying progression or reducing or inhibiting or hindering development of a neutrophil-associated condition in a subject, wherein the subject has had an inadequate clinical response to an anti-TNF therapy for the neutrophil-associated condition.

[0017] In one example, the neutrophil-associated condition is selected from the group consisting of hidradenitis suppurativa (HS); palmoplantar pustulosis (PPP); pyoderma gangrenosum; Sweet syndrome; pyoderma gangrenosum, acne, and hidradenitis suppurativa (PASH); psoriasis; rheumatoid arthritis; ankylosing spondylitis; psoriatic arthritis; Crohn’s disease; juvenile idiopathic arthritis; ulcerative colitis and combinations thereof.

[0018] In one example, the neutrophil-associated condition is HS. In another example, the neutrophil- associated condition is PPP. In a further example, the neutrophil- associated condition is pyoderma gangrenosum. In one example, the neutrophil- associated condition is Sweet syndrome. In another example, the neutrophil-associated condition is PASH. In a further example, the neutrophil- associated condition is psoriasis. In one example, the neutrophil-associated condition is rheumatoid arthritis. In another example, the neutrophil- associated condition is ankylosing spondylitis. In a further example, the neutrophil- associated condition is psoriatic arthritis. In one example, the neutrophil-associated condition is Crohn’s disease. In another example, the neutrophil- associated condition is juvenile idiopathic arthritis. In a further example, the neutrophil- associated condition is ulcerative colitis.

[0019] In one example, the neutrophil-associated condition is a neutrophil-associated autoimmune condition.

[0020] The present disclosure also provides a method of treating or delaying progression or reducing or inhibiting or hindering development of a neutrophil-associated skin condition in a subject, wherein the subject has had an inadequate clinical response to an anti-TNF therapy for the neutrophil-associated skin condition, the method comprising administering to the subject a compound that inhibits granulocyte colony stimulating factor (G-CSF) signaling.

[0021] In one example, the neutrophil-associated skin condition is selected from the group consisting of hidradenitis suppurativa (HS); pyoderma gangrenosum; Sweet syndrome; pyoderma gangrenosum, acne, hidradenitis suppurativa (PASH); psoriasis and combinations thereof.

[0022] In one example of any method described herein, prior to administering the compound the method comprises determining a G-CSF gene signature in a sample from the subject. For example, the G-CSF gene signature is determined in a peripheral blood mononuclear cells (PBMCs) sample, or a skin sample or a skin lesion sample from the subject. In one example, the G-CSF gene signature is determined in a sample of peripheral blood mononuclear cells (PBMCs) obtained from the subject. In one example, the G-CSF gene signature is determined in a sample obtained from a site of inflammation in the subject. In one example, the G-CSF gene signature is determined in a skin sample obtained from the subject. In another example, the G-CSF gene signature is determined in a skin lesion sample obtained from the subject. In one example, the method further comprises acquiring a sample of PBMCs from the subject.

[0023] In one example, the subject has a G-CSF gene signature. For example, the subject has an enriched G-CSF gene signature. In one example, the subject has an enriched G- CSF gene signature compared to a G-CSF gene signature in a control. For example, the control is a sample obtained from a healthy subject, i.e., a subject that is not suffering from a neutrophil-associated condition. In another example, the control is a sample obtained from a non-inflamed site of the subject, for example, a region of non-inflamed skin.

[0024] In one example, the subject has a positive G-CSF gene enrichment score. For example, the subject has a G-CSF gene enrichment score of >0.00 as determined by sample-level enrichment analysis. In one example, the sample-level enrichment analysis is performed using R package gene set variation analysis (GSVA).

[0025] In one example, the subject has had an inadequate clinical response to at least 12 weeks of treatment with the anti-TNF therapy. Methods of determining clinical response to treatment with an anti-TNF therapy will be apparent to the skilled person and / or described herein. For example, methods include measuring clinical scores and / or physical characteristics (e.g., skin lesions) associated with the condition being treated.

[0026] In one example, the anti-TNF therapy is an anti-TNF antibody or a soluble TNF receptor-fusion protein. In one example, the anti-TNF therapy is adalimumab, infliximab, certolizumab, golimumab or etanercept. In one example, the anti-TNF therapy is an anti- TNFa antibody. For example, the anti-TNFa antibody is adalimumab, infliximab, certolizumab or golimumab. In another example, the anti-TNF therapy is a soluble TNF receptor-fusion protein, for example, a TNF receptor-Ig fusion protein. In one example, the TNF receptor-Ig fusion protein is etanercept.

[0027] In one example, the subject has had an inadequate response, or demonstrated intolerance to, or had a contraindication to, at least 3 months of oral antibiotics for the treatment of the neutrophil-associated skin condition.

[0028] Methods of determining an inadequate response in this context, may be assessed in the same way as an inadequate response to an anti-TNF therapy as described above. In one example, administration of the compound does not cause grade 3 neutropenia or grade 4 neutropenia (or severe neutropenia) in the subject for greater than three consecutive days. In one example, administration of the compound does not cause grade 3 neutropenia or grade 4 neutropenia (or severe neutropenia) in the subject for greater than four or five or six consecutive days. In one example, administration of the compound does not cause grade 3 neutropenia or grade 4 neutropenia (or severe neutropenia) in the subject for greater than seven consecutive days.

[0029] In one example, administration of the compound does not induce grade 4 neutropenia following a single administration. In one example, administration of the compound does not induce grade 4 neutropenia following multiple administrations, e.g., two administrations or three administrations or four administrations or five administrations or six administrations. In one example, administration of the compound does not induce grade 4 neutropenia following at least three administrations.

[0030] In one example, administration of the compound induces grade 2 or grade 3 neutropenia for less than two consecutive days following a single administration. In one example, administration of the compound induces grade 2 or grade 3 neutropenia for less than two consecutive days following multiple administrations, e.g., two or three administrations or four administrations or five administrations or six administrations. In one example, administration of the compound induces grade 2 or grade 3 neutropenia for less than two consecutive days following at least three administrations.

[0031] In one example, administration of the compound does not cause sustained grade 3 or grade 4 neutropenia in the subject for greater than seven consecutive days.

[0032] In one example, administration of the compound:

[0033] (i) does not induce grade 4 neutropenia; or

[0034] (ii) does not induce neutropenia or the antibody induces grade 2 or grade 3 neutropenia for two consecutive days or less; or

[0035] (iii) does not induce neutropenia for more than 2 consecutive days or more than 1 day.

[0036] In one example, the neutropenia is not associated with a fever. In one example, the neutropenia is resolved without treatment. In one example, the neutropenia is not associated with an infection, e.g., a serious infection, such as a tuberculosis infection.

[0037] In one example, the compound that inhibits G-CSF signaling binds to G-CSF or to G-CSF receptor (G-CSFR). In one example, the compound that inhibits G-CSF signaling binds to G-CSF. In one example, the compound that inhibits G-CSF signaling binds to G-CSF receptor (G-CSFR).

[0038] In one example, the compound that inhibits G-CSF signaling is a protein. In one example, the compound that inhibits G-CSF signaling is a protein comprising an antibody variable region that binds to or specifically binds to G-CSFR and neutralizes G-CSF signaling. Reference herein to a protein or antibody that “binds to” G- CSFR provides literal support for a protein or antibody that “binds specifically to” G- CSFR.

[0039] In one example, the compound that inhibits G-CSF signaling is a protein comprising an antibody variable region that binds to or specifically binds to G-CSF and neutralizes G-CSF signaling. Reference herein to a protein or antibody that “binds to” G- CSF provides literal support for a protein or antibody that “binds specifically to” G-CSF.

[0040] In one example, the protein comprises a heavy chain variable region (VH). In one example, the protein comprises a light chain variable region (VL). In one example, the protein comprises a VH and a VL. In one example, the protein comprises a VH and a VL. In one example, the VH and a VL are in the same polypeptide chain. In another example, the VH and a VL are in separate polypeptide chains.

[0041] In one example, a protein described herein comprises at least a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH and VL bind to form a Fv comprising an antigen binding domain. In one examples, the compound that inhibits G-CSF signaling is a protein comprising a Fv. The skilled artisan will understand that the antigen binding domain comprises the binding site of the antibody.

[0042] In one example, the VH and the VL are in a single polypeptide chain. For example, the protein is:

[0043] (i) a single chain Fv fragment (scFv);

[0044] (ii) a dimeric scFv (di-scFv);

[0045] (iii) one of (i) or (ii) linked to a constant region of an antibody, Fc or a heavy chain constant domain (CH) 2 and / or CH3; or

[0046] (iv) one of (i) or (ii) linked to a protein that binds to an immune effector cell.

[0047] In one example, the VL and VH are in separate polypeptide chains.

[0048] For example, the protein is:

[0049] (i) a diabody;

[0050] (ii) a triabody;

[0051] (iii) a tetrabody;

[0052] (iv) a Fab;

[0053] (v) a F(ab’)2;

[0054] (vi) a Fv;

[0055] (vii) one of (i) to (vi) linked to a constant region of an antibody, Fc or a heavy chain constant domain (CH) 2 and / or CH3; or (viii) one of (i) to (vi) linked to a protein that binds to an immune effector cell.

[0056] In one example, the protein is selected from the group consisting of:

[0057] (i) a single chain Fv fragment (scFv);

[0058] (ii) a dimeric scFv (di-scFv);

[0059] (iii) a diabody;

[0060] (iv) a triabody;

[0061] (v) a tetrabody;

[0062] (vi) a Fab;

[0063] (vii) a F(ab’)2;

[0064] (viii) a Fv;

[0065] (ix) one of (i) to (viii) linked to a constant region of an antibody, Fc or a heavy chain constant domain (CH) 2 and / or CH3;

[0066] (x) one of (i) to (viii) linked to albumin, functional fragments or variants thereof or a protein (e.g., antibody or antigen binding fragment thereof) that binds to albumin; and

[0067] (xi) an antibody.

[0068] The foregoing proteins (described in the previous three lists) can also be referred to as antigen binding domains of antibodies.

[0069] In one example, the protein is an antibody. In one example, the antibody is a naked antibody. Exemplary antibodies are described in W02012171057, WO2018145206 and WO2022055334, all of which are incorporated herein by reference.

[0070] In one example, the protein is an antibody which binds to hG-CSFR expressed on the surface of a cell at an affinity of at least about 5 nM. In one example, the protein is an antibody which binds to hG-CSFR expressed on the surface of a cell at an affinity of at least about 4 nM. In one example, the protein is an antibody which binds to hG-CSFR expressed on the surface of a cell at an affinity of at least about 3 nM. In one example, the protein is an antibody which binds to hG-CSFR expressed on the surface of a cell at an affinity of at least about 2 nM. In one example, the protein is an antibody which binds to hG-CSFR expressed on the surface of a cell at an affinity of at least about 1 nM.

[0071] In one example, the protein is an antibody which inhibits G-CSF-induced proliferation of a BaF3 cell expressing hG-CSFR with an IC50 of at least about 5 nM. In one example, the protein is an antibody which inhibits G-CSF-induced proliferation of a BaF3 cell expressing hG-CSFR with an IC50 of at least about 4 nM. In one example, the protein is an antibody which inhibits G-CSF-induced proliferation of a BaF3 cell expressing hG-CSFR with an IC50 of at least about 3 nM. In one example, the protein is an antibody which inhibits G-CSF-induced proliferation of a BaF3 cell expressing hG- CSFR with an IC50 of at least about 2 nM. In one example, the protein is an antibody which inhibits G-CSF-induced proliferation of a BaF3 cell expressing hG-CSFR with an IC50 of at least about 1 nM. In one example, the protein is an antibody which inhibits G- CSF-induced proliferation of a BaF3 cell expressing hG-CSFR with an IC50 of at least about 0.5 nM.

[0072] In one example, the protein is chimeric, de-immunized, humanized, human or primatized. In one example, the protein or antibody is human.

[0073] In one example, the protein comprises an antibody variable region that competitively inhibits the binding of antibody C1.2G comprising a heavy chain variable region (VH) comprising a sequence set forth in SEQ ID NO: 4 and a light chain variable region (VL) comprising a sequence set forth in SEQ ID NO: 5 to G-CSFR.

[0074] In one example, the protein binds to an epitope comprising residues within one or two or three or four regions selected from 111-115, 170-176, 218-234 and / or 286-300 of SEQ ID NO: 1.

[0075] In one example, the protein is an antibody comprising a heavy chain variable region (VH) comprising an amino acid sequence set forth in SEQ ID NO: 4 and a light chain variable region (VL) comprising an amino acid sequence set forth in SEQ ID NO: 5.

[0076] In one example, the protein is an antibody comprising a VH comprising an amino acid sequence set forth in SEQ ID NO: 2 and a VL comprising an amino acid sequence set forth in SEQ ID NO: 3.

[0077] In one example, the protein is an antibody comprising a VH comprising three CDRs of a VH comprising an amino acid sequence set forth in SEQ ID NO: 4 and a VL comprising three CDRs of a VL comprising an amino acid sequence set forth in SEQ ID NO: 5.

[0078] In one example, the protein is an antibody comprising a VH comprising three CDRs of a VH comprising an amino acid sequence set forth in SEQ ID NO: 2 and a VL comprising three CDRs of a VL comprising an amino acid sequence set forth in SEQ ID NO: 3.

[0079] In one example, the CDRs are positioned according to the numbering system of Kabat.

[0080] In one example, the compound that inhibits G-CSF signaling is a protein comprising an antigen binding domain of an antibody, wherein the antigen binding domain comprises a VH and a VL, wherein:

[0081] (i) the VH comprises: a.a CDR1 comprising a sequence set forth between amino acids 31-35 of SEQ

[0082] ID NO: 2; b. a CDR2 comprising a sequence set forth between amino acids 50-65 of SEQ ID NO: 2; and c.a CDR3 comprising a sequence set forth between amino acids 99-107 of

[0083] SEQ ID NO: 2; and

[0084] (ii)the VL comprises: a.a CDR1 comprising a sequence set forth between amino acids 24-34 of SEQ

[0085] ID NO: 3; b. a CDR2 comprising a sequence set forth between amino acids 51-56 of SEQ ID NO: 3; and c.a CDR3 comprising a sequence set forth between amino acids 89-97 of SEQ

[0086] ID NO: 3.

[0087] In one example, the compound that inhibits G-CSF signaling is a protein comprising an antigen binding domain of an antibody, wherein the antigen binding domain comprises a VH and a VL, wherein:

[0088] (i) the VH comprises: a.a CDR1 comprising a sequence set forth between amino acids 31-35 of SEQ

[0089] ID NO: 4; b. a CDR2 comprising a sequence set forth between amino acids 50-65 of SEQ ID NO: 4; and c.a CDR3 comprising a sequence set forth between amino acids 99-107 of

[0090] SEQ ID NO: 4; and

[0091] (ii)the VL comprises: a.a CDR1 comprising a sequence set forth between amino acids 24-34 of SEQ

[0092] ID NO: 5; b. a CDR2 comprising a sequence set forth between amino acids 51-56 of SEQ ID NO: 5; and c.a CDR3 comprising a sequence set forth between amino acids 89-97 of SEQ

[0093] ID NO: 5.

[0094] In one example, the compound that inhibits G-CSF signaling is a protein comprising an antigen binding domain of an antibody, wherein the antigen binding domain comprises a VH and a VL, wherein:

[0095] (i) the VH comprises: a.a CDR1 comprising a sequence set forth in SEQ ID NO: 6, b. a CDR2 comprising a sequence set forth in SEQ ID NO: 7, and c.a CDR3 comprising a sequence set forth in LGELGX1X2X3X4 (SEQ ID NO: 12), wherein:

[0096] Xi is selected from the group consisting of tryptophan, glutamine, methionine, serine, phenylalanine, glutamic acid and histidine;

[0097] X2 is an amino acid selected from the group consisting of phenylalanine, tyrosine, methionine, serine, glycine and isoleucine;

[0098] X3 is an amino acid selected from the group consisting of aspartic acid, methionine, glutamine, serine, leucine, valine, arginine and histidine; and X4 is any amino acid or an amino acid selected from the group consisting of proline, glutamic acid, alanine, leucine, phenylalanine, tyrosine, threonine, asparagine, aspartic acid, serine, glycine, arginine, and lysine; and

[0099] (ii) a light chain variable region (VL) comprising: a.a CDR1 comprising a sequence set forth in SEQID NO: 9, b. a CDR2 comprising a sequence set forth in SEQ ID NO: 10, and c.a CDR3 comprising a sequence set forth in X1X2X3X4X5X6X7X8X9, (SEQ

[0100] ID NO: 13) wherein:

[0101] Xi is an amino acid selected from the group consisting of glutamine, glutamic acid, histidine, alanine and serine;

[0102] X2 is an amino acid selected from the group consisting of glutamine, valine, phenylalanine, asparagine and glutamic acid;

[0103] X3 is an amino acid selected from the group consisting of serine and glycine; X4 is an amino acid selected from the group consisting of tryptophan, methionine, phenylalanine, tyrosine, isoleucine and leucine;

[0104] X5 is an amino acid selected from the group consisting of glutamic acid, methionine, glutamine, tryptophan, serine, valine, asparagine, glycine, alanine, arginine, histidine, tyrosine, lysine and threonine;

[0105] Xe is an amino acid selected from the group consisting of tyrosine, methionine, isoleucine and threonine;

[0106] X7 is an amino acid selected from the group consisting of proline, alanine, histidine, glycine and lysine;

[0107] Xs is an amino acid selected from the group consisting of leucine, glutamine, methionine, alanine, phenylalanine, isoleucine, lysine, histidine and glycine; and X9 is an amino acid selected from the group consisting of threonine, phenylalanine, tyrosine, methionine, lysine, serine, histidine, proline, tryptophan, isoleucine, glutamine, glycine and valine.

[0108] In one example, the compound that inhibits G-CSF signaling is a protein comprising an antigen binding domain of an antibody, wherein the antigen binding domain comprises a VH and a VL, wherein:

[0109] (i) the VH comprises: a.a CDR1 comprising a sequence set forth in SEQ ID NO: 6; b. a CDR2 comprising a sequence set forth in SEQ ID NO: 7; and c.a CDR3 comprising a sequence set forth in SEQ ID NO: 8; and

[0110] (ii)the VL comprises: a.a CDR1 comprising a sequence set forth in SEQ ID NO: 9; b. a CDR2 comprising a sequence set forth in SEQ ID NO: 10; and c.a CDR3 comprising a sequence set forth in SEQ ID NO: 11.

[0111] In one example, the compound that inhibits G-CSF signaling is a protein comprising an antigen binding domain of an antibody, wherein the antigen binding domain comprises a VH and a VL, wherein:

[0112] (A) (i) the VH comprises a CDR1 comprising a sequence set forth between amino acids 31-35 of SEQ ID NO: 2; a CDR2 comprising a sequence set forth between amino acids 50-65 of SEQ ID NO: 2; and a CDR3 comprising a sequence set forth between amino acids 99-107 of SEQ ID NO: 2; and

[0113] (ii) the VL comprises a CDR1 comprising a sequence set forth between amino acids 24-34 of SEQ ID NO: 3; a CDR2 comprising a sequence set forth between amino acids 51-56 of SEQ ID NO: 3; and a CDR3 comprising a sequence set forth between amino acids 89-97 of SEQ ID NO: 3; or

[0114] (B) (i) the VH comprises a CDR1 comprising a sequence set forth between amino acids 31-35 of SEQ ID NO: 4; a CDR2 comprising a sequence set forth between amino acids 50-65 of SEQ ID NO: 4; and a CDR3 comprising a sequence set forth between amino acids 99-107 of SEQ ID NO: 4; and

[0115] (ii) the VL comprises a CDR1 comprising a sequence set forth between amino acids 24-34 of SEQ ID NO: 5; a CDR2 comprising a sequence set forth between amino acids 51-56 of SEQ ID NO: 5; and a CDR3 comprising a sequence set forth between amino acids 89-97 of SEQ ID NO: 5; or

[0116] (C) (i) the VH comprises a CDR1 comprising a sequence set forth in SEQ ID NO: 6; a CDR2 comprising a sequence set forth in SEQ ID NO: 7; and a CDR3 comprising a sequence set forth in SEQ ID NO: 8; and (ii) the VL comprises a CDR1 comprising a sequence set forth in SEQ ID NO: 9; a CDR2 comprising a sequence set forth in SEQ ID NO: 10; and CDR3 comprising a sequence set forth in SEQ ID NO: 11.

[0117] In one example, the protein or antibody as described herein comprises a human constant region, e.g., an IgG constant region, such as an IgGi, IgGi, IgGs or IgG4 constant region or mixtures thereof. In the case of an antibody or protein comprising a VH and a VL, the VH can be linked to a heavy chain constant region and the VL can be linked to a light chain constant region.

[0118] In one example, a protein or antibody as described herein comprises a constant region of an IgG4 antibody or a stabilized constant region of an IgG4 antibody. In one example, the protein or antibody comprises an IgG4 constant region with a proline at position 241 (according to the numbering system of Kabat (Kabat et al., Sequences of Proteins of Immunological Interest Washington DC United States Department of Health and Human Services, 1987 and / or 1991)).

[0119] The C-terminal lysine of the heavy chain constant region of a whole antibody (or a protein or antibody comprising a constant region or a CH3) of the disclosure may be removed, for example, during production or purification of the antibody or protein, or by recombinantly engineering the nucleic acid encoding a heavy chain of the antibody. Accordingly, whole antibodies may comprise populations with all C-terminal lysine residues removed, populations with no C-terminal lysine residues removed, and / or populations having a mixture of protein with and without the C-terminal lysine residue. In some examples, the populations may additionally comprise protein in which the C- terminal lysine residue is removed in one of the heavy chain constant regions. Similarly, a composition of whole antibodies may comprise the same or a similar mix of antibody populations with or without the C-terminal lysine residue.

[0120] In one example a protein or antibody as described herein or a composition of a protein or antibody as described herein, comprises a heavy chain constant region, including a stabilized heavy chain constant region, comprising a mixture of sequences fully or partially with or without the C-terminal lysine residue.

[0121] In one example, an antibody of the disclosure comprises a VH disclosed herein linked or fused to an IgG4 constant region or stabilized IgG4 constant region (e.g., as discussed above) and the VL is linked to or fused to a kappa light chain constant region.

[0122] In one example, the protein is an antibody comprising:

[0123] (i) a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 14 or 18 and a light chain comprising an amino acid sequence set forth in SEQ ID NO: 15; or (ii) one heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 14 and one heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 18 and two light chains comprising an amino acid sequence set forth in SEQ ID NO: 15.

[0124] In one example, the protein is an antibody comprising a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 14 or 18 and a light chain comprising an amino acid sequence set forth in SEQ ID NO: 15. For example, the protein is an antibody comprising a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 14 and a light chain comprising an amino acid sequence set forth in SEQ ID NO: 15. In another example, the protein is an antibody comprising a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 18 and a light chain comprising an amino acid sequence set forth in SEQ ID NO: 15.

[0125] In one example, the protein is an antibody comprising one heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 14 and one heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 18 and two light chains comprising an amino acid sequence set forth in SEQ ID NO: 15.

[0126] In one example, the protein or antibody is any form of the protein or antibody encoded by a nucleic acid encoding any of the foregoing proteins or antibodies.

[0127] In one example, the compound that inhibits G-CSF signaling is a protein comprising an antibody variable region, wherein the antibody variable region comprises a VH expressed by a nucleic acid encoding an amino acid of SEQ ID NO: 4 and a VL expressed by a nucleic acid encoding an amino acid of SEQ ID NO: 5.

[0128] In one example, the compound that inhibits G-CSF signaling is a protein comprising an antibody variable region, wherein the antibody variable region comprises a VH expressed by a nucleic acid encoding an amino acid of SEQ ID NO: 2 and a VL expressed by a nucleic acid encoding an amino acid of SEQ ID NO: 3.

[0129] In one example, the compound that inhibits G-CSF signaling is a protein comprising an antibody variable region, wherein the antibody variable region comprises:

[0130] (i) a VH comprising:

[0131] (a) a CDR1 expressed by a nucleic acid encoding an amino acid of SEQ ID NO: 6;

[0132] (b) a CDR2 expressed by a nucleic acid encoding an amino acid of SEQ ID NO: 7; and

[0133] (c) a CDR3 expressed by a nucleic acid encoding an amino acid of SEQ ID NO: 8; and

[0134] (ii) a VL comprising: (a) a CDR1 expressed by a nucleic acid encoding an amino acid of SEQ ID NO: 9;

[0135] (b) a CDR2 expressed by a nucleic acid encoding an amino acid of SEQ ID NO: 10; and

[0136] (c) a CDR3 expressed by a nucleic acid encoding an amino acid of SEQ ID NO: 11.

[0137] In one example, the compound that inhibits G-CSF signaling is a protein comprising an antibody variable region, wherein the antibody variable region comprises:

[0138] (i) a VH comprising:

[0139] (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 23;

[0140] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 24; and

[0141] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 25; and

[0142] (ii) a VL comprising:

[0143] (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 26;

[0144] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 27; and

[0145] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 28.

[0146] In one example, the protein or antibody comprises:

[0147] (i) a VH comprising:

[0148] (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 23;

[0149] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 24; and

[0150] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 25; and

[0151] (ii) a VL comprising:

[0152] (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 26;

[0153] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 27; and (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 28.

[0154] In one example, the compound that inhibits G-CSF signaling is a protein comprising an antibody variable region, wherein the antibody variable region comprises a VH comprising three CDRs of a VH comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 21 and a VL comprising three CDRs of a VL comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 22.

[0155] In one example, the compound that inhibits G-CSF signaling is a protein comprising an antibody variable region, wherein the antibody variable region comprises a VH comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 21; and a VL comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 22.

[0156] In one example, the compound that inhibits G-CSF signaling is a protein comprising an antibody variable region, wherein the antibody variable region comprises a heavy chain comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 19; and a light chain comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 20.

[0157] The functional characteristics of a compound that inhibits G-CSF signaling of the disclosure will be taken to apply mutatis mutandis to an antibody of the disclosure.

[0158] An exemplary method disclosed herein is a method of treating or delaying progression or reducing or inhibiting or hindering development of a neutrophil- associated condition in a subject, wherein the subject has had an inadequate clinical response to adalimumab for the neutrophil- associated condition, the method comprising administering to the subject anumigilimab.

[0159] An exemplary method disclosed herein is a method of treating or delaying progression or reducing or inhibiting or hindering development of a neutrophil- associated skin condition in a subject, wherein the subject has had an inadequate clinical response to adalimumab for the neutrophil-associated skin condition, the method comprising administering to the subject anumigilimab.

[0160] The present disclosure also provides a kit comprising a compound that inhibits G- CSF signaling packaged with instructions for use in treating or delaying progression or reducing or inhibiting or hindering development of a neutrophil-associated condition in a subject, wherein the subject has had an inadequate clinical response to an anti-TNF therapy for the neutrophil-associated condition as described herein.

[0161] In one example, the kit optionally comprises an additional therapy for administration in use in combination with a compound of the disclosure. In one example, the subject is a human. In one example, the subject is an adult, for example over 18 years of age. In one example, the subject is a child, for example less than 18 years of age.

[0162] BRIEF DESCRIPTION OF THE DRAWINGS

[0163] Figure 1 is a heatmap of the in-house generated G-CSF gene signature in the CSL324_1001 RNA sequencing data. Darker shading indicates increased expression of genes at Day 3 (diagonal striped top bar) relative to the pre-treatment (vertical striped top bar). Lighter shaded columns indicate samples from CSL324-treated patients and associated decrease in transcriptional activity indicative of CSL324 effect on GCSF signaling.

[0164] Figure 2 is an analysis of gene expression in subjects from the PIONEER clinical trial. White dots indicate skin samples taken from HS individuals that did not respond to adalimumab in the PIONEER studies. Black dots indicate samples from HS individuals considered responders. Diamonds indicate samples from healthy individuals. In the PIONEER studies, response to adalimumab was defined at week 12 using the metric of achievement of Hidradenitis Suppurativa Clinical Response (HiSCR), which is a 50% reduction in overall abscess and nodule count with no increase in abscess count and no increase in draining fistula count. Wilcoxon Rank Sum test was applied to assess the difference between the groups as indicated. ****p<0.0001, *p<0.05.

[0165] Figure 3 is an assessment of the enrichment score of (A) Thl7 cells gene sets and (B-C) IL- 17 gene sets on the PIONEER skin samples. White dots indicate skin samples taken from HS individuals that did not respond to adalimumab in the PIONEER studies. Black dots indicate samples from HS individuals considered responders. Diamonds indicate samples from healthy individuals. Wilcoxon Rank Sum test was applied to assess the difference between the groups as indicated. ****p<0.0001, ns non- significant.

[0166] Figure 4 is the result from running fGSEA with ClusterProfiler to compare the gene expression profile of HS skin lesions to skin from normal individuals.

[0167] Figure 5 is an assessment of the enrichment score of top 3 pathways identified as activated in HS kin compared with normal skin on the PIONEER skin samples: (A) Interferon alpha (B) Interferon gamma and (C) IL6 / JAK / STAT3. White dots indicate skin samples taken from HS individuals that did not respond to adalimumab in the PIONEER studies. Black dots indicate samples from HS individuals considered responders. Diamonds indicate samples from healthy individuals. Wilcoxon Rank Sum test was applied to assess the difference between the groups as indicated. ****p<0.0001, *p<0.05, ns non-significant. KEY TO SEQUENCE LISTING

[0168] SEQ ID NO: 1 amino acids 25-335 of Homo sapiens G-CSFR (hG-CSFR) with a

[0169] C-terminal polyhistidine tag

[0170] SEQ ID NO: 2 VH of Cl.2

[0171] SEQ ID NO: 3 VL 0f C1.2

[0172] SEQ ID NO: 4 VH of C1.2G

[0173] SEQ ID NO: 5 VLof C1.2G

[0174] SEQ ID NO: 6 HCDR1 of C 1.2

[0175] SEQ ID NO: 7 HCDR2 of C 1.2

[0176] SEQ ID NO: 8 HCDR3 of C 1.2

[0177] SEQ ID NO: 9 LCDR1 of Cl.2

[0178] SEQ ID NO: 10 LCDR2 of C1.2

[0179] SEQ ID NO: 11 LCDR3 of Cl.2

[0180] SEQ ID NO: 12 consensus sequence of HCDR3 of Cl.2

[0181] SEQ ID NO: 13 consensus sequence of LCDR3 of Cl.2

[0182] SEQ ID NO: 14 Heavy chain of C1.2G with stabilized IgG4 constant region

[0183] SEQ ID NO: 15 Light chain of C1.2G with kappa constant region

[0184] SEQ ID NO: 16 sequence of exemplary h-G-CSFR

[0185] SEQ ID NO: 17 polypeptide comprising Ig and CRH domains of Macaca fascicularis G-CSFR (cynoG-CSFR) with a C-terminal polyhistidine tag

[0186] SEQ ID NO: 18 Heavy chain of C1.2G with stabilized IgG4 constant region and lacking C-terminal lysine

[0187] SEQ ID NO: 19 nucleotide sequence of heavy chain of C1.2G

[0188] SEQ ID NO: 20 nucleotide sequence of light chain of C1.2G

[0189] SEQ ID NO: 21 nucleotide sequence of VH of C1.2G

[0190] SEQ ID NO: 22 nucleotide sequence of VL of C1.2G

[0191] SEQ ID NO: 23 nucleotide sequence of HCDR1 of C1.2G

[0192] SEQ ID NO: 24 nucleotide sequence of HCDR2 of C1.2G

[0193] SEQ ID NO: 25 nucleotide sequence of HCDR3 of C1.2G

[0194] SEQ ID NO: 26 nucleotide sequence of LCDR1 of C1.2G

[0195] SEQ ID NO: 27 nucleotide sequence of LCDR2 of C1.2G

[0196] SEQ ID NO: 28 nucleotide sequence of LCDR3 of C1.2G

[0197] DESCRIPTION General

[0198] Throughout this specification, unless specifically stated otherwise or the context requires otherwise, reference to a single step, composition of matter, group of steps or group of compositions of matter shall be taken to encompass one and a plurality (i.e. one or more) of those steps, compositions of matter, groups of steps or groups of compositions of matter.

[0199] Those skilled in the art will appreciate that the present disclosure is susceptible to variations and modifications other than those specifically described. It is to be understood that the disclosure includes all such variations and modifications. The disclosure also includes all of the steps, features, compositions and compounds referred to or indicated in this specification, individually or collectively, and any and all combinations or any two or more of said steps or features.

[0200] The present disclosure is not to be limited in scope by the specific examples described herein, which are intended for the purpose of exemplification only. Functionally-equivalent products, compositions and methods are clearly within the scope of the present disclosure.

[0201] Any example of the present disclosure herein shall be taken to apply mutatis mutandis to any other example of the disclosure unless specifically stated otherwise. Stated another way, any specific example of the present disclosure may be combined with any other specific example of the disclosure (except where mutually exclusive).

[0202] Any example of the present disclosure disclosing a specific feature or group of features or method or method steps will be taken to provide explicit support for disclaiming the specific feature or group of features or method or method steps.

[0203] Unless specifically defined otherwise, all technical and scientific terms used herein shall be taken to have the same meaning as commonly understood by one of ordinary skill in the art (for example, in cell culture, molecular genetics, immunology, immunohistochemistry, protein chemistry, and biochemistry).

[0204] Unless otherwise indicated, the recombinant protein, cell culture, and immunological techniques utilized in the present disclosure are standard procedures, well known to those skilled in the art. Such techniques are described and explained throughout the literature in sources such as, J. Perbal, A Practical Guide to Molecular Cloning, John Wiley and Sons (1984), J. Sambrook et al. Molecular Cloning: A Laboratory Manual, Cold Spring Harbour Laboratory Press (1989), T.A. Brown (editor), Essential Molecular Biology: A Practical Approach, Volumes 1 and 2, IRL Press (1991), D.M. Glover and B.D. Hames (editors), DNA Cloning: A Practical Approach, Volumes 1-4, IRL Press (1995 and 1996), and F.M. Ausubel et al. (editors), Current Protocols in Molecular Biology, Greene Pub. Associates and Wiley-Interscience (1988, including all updates until present), Ed Harlow and David Lane (editors) Antibodies: A Laboratory Manual, Cold Spring Harbour Laboratory, (1988), and J.E. Coligan et al. (editors) Current Protocols in Immunology, John Wiley & Sons (including all updates until present).

[0205] The description and definitions of variable regions and parts thereof, antibodies and fragments thereof herein may be further clarified by the discussion in Kabat Sequences of Proteins of Immunological Interest, National Institutes of Health, Bethesda, Md., 1987 and 1991.

[0206] The term “EU numbering system of Kabat” will be understood to mean the numbering of an antibody heavy chain is according to the EU index as taught in Kabat et al., 1991, Sequences of Proteins of Immunological Interest, 5th Ed., United States Public Health Service, National Institutes of Health, Bethesda. The EU index is based on the residue numbering of the human IgGl EU antibody.

[0207] Any discussion of a protein or antibody herein will be understood to include any variants of the protein or antibody produced during manufacturing and / or storage. For example, during manufacturing or storage an antibody can be deamidated (e.g., at an asparagine or a glutamine residue) and / or have misincorporated amino acid residues (e.g., a serine misincorporated in place of an asparagine residue) and / or have altered glycosylation and / or have a glutamine residue converted to pyroglutamine and / or have a N-terminal or C-terminal residue removed or “clipped” and / or have part or all of a signal sequence incompletely processed and, as a consequence, remain at the terminus of the antibody. It is understood that a composition comprising a particular amino acid sequence may be a heterogeneous mixture of the stated or encoded sequence and / or variants of that stated or encoded sequence.

[0208] The term “and / or”, e.g., “X and / or Y” shall be understood to mean either “X and Y” or “X or Y” and shall be taken to provide explicit support for both meanings or for either meaning.

[0209] Throughout this specification the word “comprise”, or variations such as “comprises” or “comprising”, will be understood to imply the inclusion of a stated element, integer or step, or group of elements, integers or steps, but not the exclusion of any other element, integer or step, or group of elements, integers or steps.

[0210] As used herein the term “derived from” shall be taken to indicate that a specified integer may be obtained from a particular source albeit not necessarily directly from that source.

[0211] All publications cited herein are hereby incorporated by reference in their entirety. Where reference is made to a URL or other such identifier or address, it is understood that such identifiers can change and particular information on the internet can come and go, but equivalent information can be found by searching the internet. Reference thereto evidences the availability and public dissemination of such information.

[0212] Any discussion of documents, acts, materials, devices, articles or the like that has been included in the present specification is solely for the purpose of providing a context for the present invention. It is not to be taken as an admission that any or all of these matters form part of the prior art base or were common general knowledge in the field relevant to the present invention as it existed before the priority date of each claim of this application.

[0213] Selected Definitions

[0214] A “compound”, as contemplated by the present disclosure, can take any of a variety of forms including natural compounds, chemical small molecule compounds or biological compounds or macromolecules. Exemplary compounds include an antibody or a protein comprising an antigen binding fragment of an antibody, a nucleic acid, a polypeptide, a peptide, and a small molecule.

[0215] Reference herein to “granulocyte colony-stimulating factor” (G-CSF) includes native forms of G-CSF, mutant forms thereof, e.g., filgrastim and pegylated forms of G- CSF or filgrastim. This term also encompasses mutant forms of G-CSF retaining activity to bind to G-CSFR (e.g., human G-CSFR) and induce signaling.

[0216] G-CSF is a major regulator of granulocyte production. G-CSF is produced by bone marrow stromal cells, endothelial cells, macrophages, and fibroblasts, and production is induced by inflammatory stimuli. G-CSF acts through the G-CSF receptor (G-CSFR), which is expressed on early myeloid progenitors, mature neutrophils, monocytes / macrophages, T and B lymphocytes and endothelial cells.

[0217] For the purposes of nomenclature only and not limitation, an exemplary sequence of a human G-CSFR is set out in NCBI Reference Sequence: NP_000751.1 (and set out in SEQ ID NO: 16). The sequence of G-CSFR from other species can be determined using sequences provided herein and / or in publically available databases and / or determined using standard techniques (e.g., as described in Ausubel et al., (editors), Current Protocols in Molecular Biology, Greene Pub. Associates and Wiley-Interscience (1988, including all updates until present) or Sambrook et al., Molecular Cloning: A Eaboratory Manual, Cold Spring Harbor Laboratory Press (1989)) Reference to human G-CSFR may be abbreviated to hG-CSFR and reference to cynomolgus monkey G- CSFR may be abbreviated to cynoG-CSFR. Reference to soluble G-CSFR refers to polypeptides comprising the ligand binding region of G-CSFR. The Ig and CRH domains of the G-CSFR are involved in ligand binding and receptor dimerization (Layton et al., J. Biol Chem., 272: 29735-29741, 1997 and Fukunaga et al, EMBO J. 10: 2855- 2865, 1991). Soluble forms of G-CSFR comprising these portions of the receptor have been used in various studies of the receptor and mutation of the free cysteines at positions 78, 163, and 228 of the receptor assists in expression and isolation of the soluble receptor polypeptide (Mine et al., Biochem., 43: 2458-2464 2004) without affecting ligand binding.

[0218] As used herein, the term “G-CSF signaling” refers to biological activities mediated via the G-CSF receptor (G-CSFR). It will be understood that reference to inhibiting G-CSF signaling encompasses inhibition of G-CSF activity, including downstream pathways, mediated via the receptor. It will be apparent to the skilled person from the disclosure herein that the compound, for example, binds the G-CSF receptor and displaces or hinders the binding of G-CSF to the receptor. For example, the G-CSF signaling is G-CSF receptor mediated signaling.

[0219] As used herein, the term “delaying progression of’ shall be understood to include administering a compound described herein to thereby reduce or hinder or slow the progression of at least one specified disease or condition, or associated symptom thereof.

[0220] As used herein, the phrase “reduce” or “reducing” shall be understood to include administering a compound described herein to make smaller or less in amount, degree or size at least one specified disease or condition, or symptom thereof.

[0221] As used herein, the phrase “inhibit” or “inhibiting” shall be understood to include administering a compound described herein to eliminate or stop, at least partially the development of at least one specified disease or condition, or symptom thereof.

[0222] As used herein, the phrase “hinder” or “hindering” shall be understood to include administrating a compound described herein to limit or impede or hamper, at least partially the development of at least one specified disease or condition, or symptom thereof.

[0223] As used herein, the term “subject” shall be taken to mean any animal including humans, for example a mammal. Exemplary subjects include but are not limited to humans and non-human primates. For example, the subject is a human.

[0224] As used herein, the term “neutropenia” is used to refer to an absolute neutrophil count (ANC) below the lower limit of normal range, for example an ANC of less than 2000 cells / pL blood, or less than 1500 cells / pL blood, or less than 1000 cells / pL blood, for example less than 500 cells / pL blood (see Sibille et al. 2010 Br J Clin Pharmacol 70(5): 736-748). In some examples, the antibody that inhibits G-CSF signaling is administered in an amount that does not cause severe neutropenia. As used herein, the term “severe neutropenia” is used to refer to an absolute neutrophil count (ANC) of less than 1000 cells / pL blood. For the purposes of the present disclosure, the following will be used to define the grades of neutropenia:

[0225] . Grade 1: < 2.0 x 109 / L (< 2000 / mm3) and > 1.1 x 109 / L (> 1500 / mm3)

[0226] . Grade 2: < 1.5 x 109 / L (< 1500 / mm3) and > 1.0 x 109 / L (> 1000 / mm3)

[0227] • Grade 3: < 1.0 x 109 / L (< 1000 / mm3) and > 0.5 x 109 / L (> 500 / mm3)

[0228] • Grade 4: < 0.5 x 109 / L (< 500 / mm3).

[0229] The term “protein” shall be taken to include a single polypeptide chain, i.e., a series of contiguous amino acids linked by peptide bonds or a series of polypeptide chains covalently or non-covalently linked to one another (i.e., a polypeptide complex). For example, the series of polypeptide chains can be covalently linked using a suitable chemical or a disulphide bond. Examples of non-covalent bonds include hydrogen bonds, ionic bonds, Van der Waals forces, and hydrophobic interactions.

[0230] The term “polypeptide” or “polypeptide chain” will be understood from the foregoing paragraph to mean a series of contiguous amino acids linked by peptide bonds.

[0231] The term "isolated protein" or "isolated polypeptide" is a protein or polypeptide that by virtue of its origin or source of derivation is not associated with naturally- associated components that accompany it in its native state; is substantially free of other proteins from the same source. A protein may be rendered substantially free of naturally associated components or substantially purified by isolation, using protein purification techniques known in the art. By “substantially purified” is meant the protein is substantially free of contaminating agents, e.g., at least about 70% or 75% or 80% or 85% or 90% or 95% or 96% or 97% or 98% or 99% free of contaminating agents.

[0232] As used herein, the term “nucleotide sequence” or “nucleic acid sequence” will be understood to mean a series of contiguous nucleotides (or bases) covalently linked to a phosphodiester backbone.

[0233] The term “recombinant” shall be understood to mean the product of artificial genetic recombination. Accordingly, in the context of a recombinant protein comprising an antibody antigen binding domain, this term does not encompass an antibody naturally- occurring within a subject’s body that is the product of natural recombination that occurs during B cell maturation. However, if such an antibody is isolated, it is to be considered an isolated protein comprising an antibody antigen binding domain. Similarly, if nucleic acid encoding the protein is isolated and expressed using recombinant means, the resulting protein is a recombinant protein comprising an antibody antigen binding domain. A recombinant protein also encompasses a protein expressed by artificial recombinant means when it is within a cell, tissue or subject, e.g., in which it is expressed. As used herein, the term “antigen binding site” shall be taken to mean a structure formed by a protein that is capable of binding or specifically binding to an antigen. The antigen binding site need not be a series of contiguous amino acids, or even amino acids in a single polypeptide chain. For example, in a Fv produced from two different polypeptide chains the antigen binding site is made up of a series of amino acids of a VL and a VH that interact with the antigen and that are generally, however not always in the one or more of the CDRs in each variable region. In some examples, an antigen binding site is a VH or a VL or a Fv.

[0234] The skilled artisan will be aware that an “antibody” is generally considered to be a protein that comprises a variable region made up of a plurality of polypeptide chains, e.g., a polypeptide comprising a VL and a polypeptide comprising a VH. An antibody also generally comprises constant domains, some of which can be arranged into a constant region, which includes a constant fragment or fragment crystallizable (Fc), in the case of a heavy chain. A VH and a VL interact to form a Fv comprising an antigen binding region that is capable of specifically binding to one or a few closely related antigens. Generally, a light chain from mammals is either a K light chain or a light chain and a heavy chain from mammals is a, 5, a, y, or p. Antibodies can be of any type (e.g., IgG, IgE, IgM, IgD, IgA, and IgY), class (e.g., IgGi, IgGi. IgGs, IgG4, IgAi and IgAi) or subclass. The term “antibody” also encompasses humanized antibodies, primatized antibodies, human antibodies and chimeric antibodies.

[0235] The terms "full-length antibody," "intact antibody" or "whole antibody" are used interchangeably to refer to an antibody in its substantially intact form, as opposed to an antigen binding fragment of an antibody. Specifically, whole antibodies include those with heavy and light chains including an Fc region. The constant domains may be wildtype sequence constant domains (e.g., human wild-type sequence constant domains) or amino acid sequence variants thereof.

[0236] As used herein, “variable region" refers to the portions of the light and / or heavy chains of an antibody as defined herein that is capable of specifically binding to an antigen and includes amino acid sequences of complementarity determining regions (CDRs); i.e., CDR1, CDR2, and CDR3, and framework regions (FRs). Exemplary variable regions comprise three or four FRs (e.g., FR1, FR2, FR3 and optionally FR4) together with three CDRs. In the case of a protein derived from an IgNAR, the protein may lack a CDR2. VH refers to the variable region of the heavy chain. VL refers to the variable region of the light chain.

[0237] As used herein, the term "complementarity determining regions” (syn. CDRs; i.e., CDR1, CDR2, and CDR3) refers to the amino acid residues of an antibody variable region the presence of which are necessary for antigen binding. Each variable region typically has three CDR regions identified as CDR1, CDR2 and CDR3. The amino acid positions assigned to CDRs and FRs can be defined according to Kabat Sequences of Proteins of Immunological Interest, National Institutes of Health, Bethesda, Md., 1987 and 1991 or other numbering systems in the performance of this disclosure, e.g., the canonical numbering system of Chothia and Lesk J. Mol Biol. 196: 901-917, 1987; Chothia et al. Nature 342, 877-883, 1989; and / or Al-Lazikani et al., J Mol Biol 273: 927-948, 1997; the IMGT numbering system of Lefranc et al., Devel. And Compar. Immunol., 27: 55- 77 , 2003; or the AHO numbering system of Honnegher and Pliikthun J. Mol. Biol., 309: 657-670, 2001. For example, according to the numbering system of Kabat, VH framework regions (FRs) and CDRs are positioned as follows: residues 1-30 (FR1 ), 31- 35 (CDR1), 36-49 (FR2), 50-65 (CDR2), 66-94 (FR3), 95-102 (CDR3) and 103- 113 (FR4). According to the numbering system of Kabat, VL FRS and CDRs are positioned as follows: residues 1-23 (FR1), 24-34 (CDR1), 35-49 (FR2), 50-56 (CDR2), 57-88 (FR3), 89-97 (CDR3) and 98-107 (FR4). The present disclosure is not limited to FRs and CDRs as defined by the Kabat numbering system, but includes all numbering systems, including those discussed above. In one example, reference herein to a CDR (or a FR) is in respect of those regions according to the Kabat numbering system.

[0238] "Framework regions" (FRs) are those variable region residues other than the CDR residues.

[0239] As used herein, the term “Fv” shall be taken to mean any protein, whether comprised of multiple polypeptides or a single polypeptide, in which a VL and a VH associate and form a complex having an antigen binding site, i.e., capable of specifically binding to an antigen. The VH and the VL which form the antigen binding site can be in a single polypeptide chain or in different polypeptide chains. Furthermore, an Fv of the disclosure (as well as any protein of the disclosure) may have multiple antigen binding sites which may or may not bind the same antigen. This term shall be understood to encompass fragments directly derived from an antibody as well as proteins corresponding to such a fragment produced using recombinant means. In some examples, the VH is not linked to a heavy chain constant domain (CH) 1 and / or the VL is not linked to a light chain constant domain (CL). Exemplary Fv containing polypeptides or proteins include a Fab fragment, a Fab’ fragment, aF(ab’) fragment, a scFv, a diabody, atriabody, a tetrabody or higher order complex, or any of the foregoing linked to a constant region or domain thereof, e.g., CH2 or CH3 domain, e.g., a minibody. A "Fab fragment" consists of a monovalent antigen-binding fragment of an immunoglobulin, and can be produced by digestion of a whole antibody with the enzyme papain, to yield a fragment consisting of an intact light chain and a portion of a heavy chain or can be produced using recombinant means. A "Fab' fragment" of an antibody can be obtained by treating a whole antibody with pepsin, followed by reduction, to yield a molecule consisting of an intact light chain and a portion of a heavy chain comprising a VH and a single constant domain. Two Fab' fragments are obtained per antibody treated in this manner. A Fab’ fragment can also be produced by recombinant means. A "F(ab')2 fragment” of an antibody consists of a dimer of two Fab' fragments held together by two disulfide bonds, and is obtained by treating a whole antibody molecule with the enzyme pepsin, without subsequent reduction. A “Fab ” fragment is a recombinant fragment comprising two Fab fragments linked using, for example a leucine zipper or a CH3 domain. A “single chain Fv” or “scFv” is a recombinant molecule containing the variable region fragment (Fv) of an antibody in which the variable region of the light chain and the variable region of the heavy chain are covalently linked by a suitable, flexible polypeptide linker.

[0240] As used herein, the term “binds” in reference to the interaction of a compound or an antigen binding site thereof with an antigen means that the interaction is dependent upon the presence of a particular structure (e.g., an antigenic determinant or epitope) on the antigen. For example, an antibody recognizes and binds to a specific protein structure rather than to proteins generally. If an antibody binds to epitope "A", the presence of a molecule containing epitope “A” (or free, unlabeled “A”), in a reaction containing labeled “A” and the protein, will reduce the amount of labeled “A” bound to the antibody.

[0241] As used herein, the term “specifically binds” or “binds specifically” shall be taken to mean that a compound of the disclosure reacts or associates more frequently, more rapidly, with greater duration and / or with greater affinity with a particular antigen or cell expressing same than it does with alternative antigens or cells. For example, a compound binds to G-CSFR (e.g., hG-CSFR) with materially greater affinity (e.g., 20 fold or 40 fold or 60 fold or 80 fold to 100 fold or 150 fold or 200 fold) than it does to other cytokine receptor or to antigens commonly recognized by polyreactive natural antibodies (i.e., by naturally occurring antibodies known to bind a variety of antigens naturally found in humans). Generally, but not necessarily, reference to binding means specific binding, and each term shall be understood to provide explicit support for the other term.

[0242] A protein or antibody may be considered to “preferentially bind” to a polypeptide if it binds that polypeptide with a dissociation constant (KD) that is less than the protein’s or antibody's KD for another polypeptide. In one example, a protein or antibody is considered to preferentially bind to a polypeptide if it binds the polypeptide with an affinity (i.e., KD) that is at least about 20 fold or 40 fold or 60 fold or 80 fold or 100 fold or 120 fold or 140 fold or 160 fold more than the protein’s or antibody's KD for another polypeptide.

[0243] For the purposes of clarification and as will be apparent to the skilled artisan based on the exemplified subject matter herein, reference to “affinity” in this specification is a reference to KD of a protein or antibody.

[0244] For the purposes of clarification and as will be apparent to the skilled artisan based on the description herein, reference to an “affinity of at least about” will be understood to mean that the affinity (or KD) is equal to the recited value or higher (i.e., the value recited as the affinity is lower), i.e., an affinity of 2nM is greater than an affinity of 3nM. Stated another way, this term could be “an affinity of X or less”, wherein X is a value recited herein.

[0245] An “ICso of at least about” will be understood to mean that the IC50 is equal to the recited value or greater (i.e., the value recited as the IC50 is lower), i.e., an IC50 of 2nM is greater than an IC50 of 3nM. Stated another way, this term could be “an IC50 of X or less”, wherein X is a value recited herein.

[0246] As used herein, the term “epitope” (syn. “antigenic determinant”) shall be understood to mean a region of hG-CSFR to which a protein comprising an antigen binding site of an antibody binds. This term is not necessarily limited to the specific residues or structure to which the protein makes contact. For example, this term includes the region spanning amino acids contacted by the protein and / or 5-10 or 2-5 or 1-3 amino acids outside of this region. In some examples, the epitope comprises a series of discontinuous amino acids that are positioned close to one another when hG-CSFR is folded, i.e., a “conformational epitope”. For example, a conformational epitope comprises amino acids in one or more or two or more or all of the regions corresponding to 111-115, 170-176, 218-234 and / or 286-300 of SEQ ID NO: 1. The skilled artisan will also be aware that the term "epitope" is not limited to peptides or polypeptides. For example, the term “epitope” includes chemically active surface groupings of molecules such as sugar side chains, phosphoryl side chains, or sulfonyl side chains, and, in certain examples, may have specific three dimensional structural characteristics, and / or specific charge characteristics.

[0247] The term “competitively inhibits” shall be understood to mean that a protein of the disclosure (or an antigen binding site thereof) reduces or prevents binding of a recited antibody or protein to G-CSFR, e.g., to hG-CSFR. This may be due to the protein (or antigen binding site) and antibody binding to the same or an overlapping epitope. It will be apparent from the foregoing that the protein need not completely inhibit binding of the antibody, rather it need only reduce binding by a statistically significant amount, for example, by at least about 10% or 20% or 30% or 40% or 50% or 60% or 70% or 80% or 90% or 95%. Preferably, the protein reduces binding of the antibody by at least about 30%, more preferably by at least about 50%, more preferably, by at least about 70%, still more preferably by at least about 75%, even more preferably, by at least about 80% or 85% and even more preferably, by at least about 90%. Methods for determining competitive inhibition of binding are known in the art and / or described herein. For example, the antibody is exposed to G-CSFR either in the presence or absence of the protein. If less antibody binds in the presence of the protein than in the absence of the protein, the protein is considered to competitively inhibit binding of the antibody. In one example, the competitive inhibition is not due to steric hindrance.

[0248] “Overlapping” in the context of two epitopes shall be taken to mean that two epitopes share a sufficient number of amino acid residues to permit a protein (or antigen binding site thereof) that binds to one epitope to competitively inhibit the binding of a protein (or antigen binding site) that binds to the other epitope. For example, the “overlapping” epitopes share at least 1 or 2 or 3 or 4 or 5 or 6 or 7 or 8 or 9 or 20 amino acids.

[0249] As used herein, the term “neutralize” shall be taken to mean that a compound is capable of blocking, reducing or preventing G-CSF-mediated signaling in a cell through the G-CSFR. Methods for determining neutralization are known in the art and / or described herein.

[0250] Neutrophil-associated conditions

[0251] The present disclosure provides a method of treating or delaying progression or reducing or inhibiting or hindering development of a neutrophil-associated condition in a subject, wherein the subject has had an inadequate clinical response to an anti-TNF therapy. An inadequate clinical response may be assessed by a clinician after or during the anti-TNF therapy.

[0252] As used herein, the term “inadequate clinical response” is used synonymously herein with “inadequate response” and “non-responsive” and refers to an inadequate improvement in disease symptoms after treatment with the anti-TNF therapy.

[0253] In one example, the neutrophil- associated condition is an autoimmune disease or a neutrophilic skin condition, e.g., a neutrophilic dermatosis or a neutrophilic skin lesion.

[0254] Neutrophil-associated autoimmune conditions

[0255] In one example, the neutrophil-associated condition is an autoimmune disease. Exemplary autoimmune conditions will be apparent to the skilled person and / or include autoimmune intestinal disorders (such as Crohn’s disease and ulcerative colitis), arthritis (such as rheumatoid arthritis, psoriatic arthritis or idiopathic arthritis, e.g., juvenile idiopathic arthritis) or psoriasis.

[0256] In one example, the neutrophil- associated condition is an autoimmune intestinal disorder. For example, the condition is Crohn’s disease. In another example, the condition is ulcerative colitis.

[0257] The present disclosure also provides methods for improving Crohn’s symptoms in a subject based on indices used to measure the disease state. Methods of measuring disease state and / or determining response to an anti-TNF therapy and / or compound described herein will be apparent to the skilled person and / or described herein. For example, subjects with Crohn's disease can be evaluated using the Crohn's Disease Activity Index (CD Al), which is a standard measure of the severity of the disease based on number of stools, abdominal pain, general well-being, extraintestinal complications, antidiarrheal agents used in the previous 7 days, abdominal mass felt on palpation, hematocrit, and body weight. CD Al scores range from 0 to 600. A score of less than 150 corresponds to relative disease quiescence (remission); 150 to 219, mildly active disease; 220 to 450, moderately active disease; and greater than 450, severe disease. A decrease in greater than 100 points indicates a clinically significant improvement in disease activity.

[0258] The present disclosure also provides methods for improving ulcerative colitis symptoms in a subject based on indices used to measure the disease state. Methods of measuring disease state and / or determining response to an anti-TNF therapy and / or a compound described herein will be apparent to the skilled person and / or described herein. For example, subjects with ulcerative colitis can be evaluated using the Ulcerative Colitis Endoscopic Index of Severity (UCEIS) score. In the UCEIS strata, UCEIS 0-1 = remission; UCEIS 2-4 = mild disease; UCEIS 5-6 = moderate disease; UCEIS 7-8 = severe ulcerative colitis.

[0259] In one example, the neutrophil-associated condition is rheumatoid arthritis, psoriatic arthritis or idiopathic arthritis, e.g., juvenile idiopathic arthritis.

[0260] In one example, the neutrophil- associated condition is rheumatoid arthritis (RA). Certain subtypes of RA may be treated in accordance with the present disclosure. In one example, moderate to severe RA, is treated by administering a compound disclosed herein. In one example, the subject has confirmed moderate to severe RA, e.g., polyarticular RA.

[0261] The present disclosure also provides methods for improving RA symptoms in a subject based on indices used to measure the disease state. Methods for measuring the severity of RA and for determining response to an anti-TNF therapy and / or a compound described herein will be apparent to the skilled person and / or described herein. For example, comparing the number of tender and swollen joints between baseline and various time points during treatment is a typical way to assess joint status and response to treatment. In the American College of Rheumatology (ACR) joint count for RA (Felson et al. Arthritis & Rheumatology 38: 727-735, 1995), 68 joints are assessed for tenderness and 66 for swelling (the hip is not assessed for swelling). In the Disease Activity Score (DAS) employed primarily in Europe, either a 44- or 28-joint count is used in RA. In addition to the joint count, the ACR evaluation criteria include the following elements to comprise a composite score: patient global (on a visual analog scale [VAS]), patient pain, physician global, Health Assessment Questionnaire (HAQ; a measure of function), and an acute-phase reactant (either C-reactive protein or sedimentation rate). An ACR 20 response would constitute a 20% improvement in tender and swollen joint count and a 20% improvement of at least 3 of the other 5 elements in the composite criteria. ACR 50 and 70 responses represent at least a 50% and 70% improvement of these elements. The ACR system only represents change, whereas the DAS system represents both current state of disease activity and change. The DAS scoring system uses a weighted mathematical formula, derived from clinical trials in RA. For example, the DAS 28 is 0.56(T28)+0.28( SW28)+0.70(Ln ESR)+0.014 GH wherein T represents tender joint number, SW is swollen joint number, ESR is erythrocyte sedimentation rate, and GH is global health. Various values of the DAS represent high or low disease activity as well as remission, and the change and endpoint score result in a categorization of the patient by degree of response (none, moderate, good).

[0262] In one example, the neutrophil-associated condition is juvenile idiopathic arthritis. The term “juvenile rheumatoid arthritis” or “JRA” as used herein refers to a chronic, inflammatory disease which occurs before age 16 that may cause joint or connective tissue damage. JRA is also referred to as juvenile chronic polyarthritis and Still's disease.

[0263] Certain subtypes of JRA may be treated in accordance with the present disclosure, including pauciarticular JRA, polyarticular JRA or systemic JRA. In one example, moderate to severe JRA, is treated by administering a compound disclosed herein. In one example, the subject has confirmed moderate to severe JRA, e.g., polyarticular RA.

[0264] The present disclosure also provides methods for improving JRA symptoms in a subject based on indices used to measure the disease state. Methods for measuring the severity of JRA and for determining response to an anti-TNF therapy and / or a compound described herein will be apparent to the skilled person and / or described herein. For example, subjects with JRA can be evaluated using the physician’s global assessment of disease activity (PhGA), the Juvenile Arthritis Disease Activity Scores (J AD AS 10), and the Clinical (cJADASlO) and Systemic (sJADAS) variations. The JADAS10 includes the following 4 measures: physician global assessment of disease activity (PhGA) measured on a 0-10 visual analog scale (VAS), parent / patient global assessment of child well-being measured on a 0-10 VAS, count of 10 joints with active disease among the total assessed, and the erythrocyte sedimentation rate (ESR) or C-reactive protein (CRP) level, both normalized to a 0-10 scale. Cutoff values separate states of inactive disease (ID), minimal disease activity (MiDA), moderate disease activity (MoDA), and high disease activity (HDA) with cutoff scores shown in Table 1.

[0265] (Trincianti et al., Arthritis Rheumatol. 2021 Nov; 73(11): 1966-1975.)

[0266] Neutrophil -associated skin conditions

[0267] The present disclosure relates to methods of treating neutrophilic or neutrophil- associated skin conditions. As used herein, the term “neutrophilic skin condition” or “neutrophil-associated skin condition” refers to any disease or condition which is characterised by one or more lesions with intense epidermal, dermal, or hypodermal infiltrates composed primarily of neutrophils, with no evidence of infection or true vasculitis. This includes a “neutrophilic dermatosis”, i.e., a disease of the skin, as well as a “neutrophilic skin lesion”, which refers to a lesion for which the underlying disease may not have been diagnosed, but which is characterised by neutrophil infiltration as described above.

[0268] Exemplary neutrophil-associated skin condition includes hidradenitis suppurativa (HS), palmoplantar pustulosis (PPP); pyoderma gangrenosum; Sweet syndrome; pyoderma gangrenosum, acne, hidradenitis suppurativa (PASH); and psoriasis.

[0269] In one example, the neutrophil-associated skin condition is of low prevalence. For example, the low prevalence neutrophil-associated skin condition is selected from the group consisting of HS, PASH, pyoderma gangrenosum, PPP and combinations thereof. In some examples, the neutrophil- associated skin condition is hidradenitis suppurativa (HS). HS is a skin disorder of the apocrine glands (sweat glands found on certain parts of the body) and hair follicles in which swollen, painful, inflamed lesions or lumps develop in the groin and sometimes under the arms and under the breasts. HS occurs when apocrine gland outlets become blocked by perspiration or are unable to drain normally because of incomplete gland development. Secretions trapped in the glands force perspiration and bacteria into surrounding tissue, causing subcutaneous induration, inflammation, and infection. HS is confined to areas of the body that contain apocrine glands. These areas are the axillae, areola of the nipple, groin, perineum, circumanal, and periumbilical regions.

[0270] The present disclosure also provides a method for treating certain subpopulations of HS patients who may be especially difficult to treat, i.e., those who have had inadequate clinical response to an anti-TNF therapy.

[0271] The present disclosure provides a method for treating moderate to severe HS.

[0272] The present disclosure also provides a method for treating moderate to severe HS in adolescents 12 years and older.

[0273] Methods of determining severity of disease and / or response to an anti-TNF therapy and / or a compound described herein will be apparent to the skilled person and / or described herein. For example, treatment of HS (or response) may be determined using any of the measures known in the art, e.g., improvement in Hurley Staging or the Sartorius scale, or any measure known to those in the art. For example, in one instance, an improvement in the Hurley stage of the subject having HS, or any of the measures described herein, is evidence of effective HS treatment. In one instance, the severity of HS is determined according to the Hurley staging system. Hurley staging is based on assigning the subject having HS one of three different "Stages" depending on the disease level. More specifically, Stage I refers to abscess formation, single or multiple, without sinus tracts and cicatrisation; Stage II refers to recurrent abscesses with tract formation and cicatrisation, as well as single or multiple, widely separated lesions; and Stage III, which refers to diffuse or near-diffuse involvement, or multiple interconnected tracts and abscesses across the entire area. Hurley Stage III is the most severe form. In one instance, the subject having HS has HS lesions that are present in at least two distinct anatomic areas (e.g.left and right axilla; or left axilla and left inguinal-crural fold), one of which is at least Hurley Stage II. In another instance, the subject being treated has at least one lesion that is at least a Hurley Stage II.

[0274] In one example, treatment of HS with compound disclosed herein is determined by an improved Hurley score relative to a given baseline, e.g., the Hurley stage of the subject prior to treatment with the compound. In one example, improvement in a Hurley score indicates that the Hurley score of the subject has either improved or been maintained following treatment with a compound.

[0275] Severity of HS may be determined according to standard clinical definitions. See, for example, Hurley staging {III vs. (I or II)} for HS (Poli F, Jemec GBE, Revuz J., Clinical Presentation. In: Jemec GBE, Revuz J, Leyden JJ, editors. Hidradenitis Suppurativa. Springer, New York, 2006, pp 11-24 ). Hurley stage III disease is the most severe stage of hidradenitis suppurativa, reflecting diffuse or near-diffuse involvement of affected areas.

[0276] In another example, severity of HS disease can be determined according to the International Hidradenitis Suppurativa Severity Score System (IHS4). The IHS4 is a validated tool for the dynamic severity assessment of HS (Zouboulis, et al., Br J Dermatol, 177 1401-09, 2017) and is designed to assess treatment response rather than disease severity cross-sectionality (Kimball et al., Br J Dermatol, 777: 1434-42, 2014). The IHS4 score (points) = (number of nodules multiplied by 1) + (number of abscesses multiplied by 2) + [number of draining tunnels (fistulae / sinuses) multiplied by 4]. A score of 3 or less signifies mild HS, a score of 4-10 signifies moderate HS and a score of 11 or higher signifies severe HS (Zouboulis, et al., Br J Dermatol, 177 1401-09, 2017). In some examples, the subject has an IHS4 score of > 4 prior to treatment.

[0277] In some examples, the Sartorius scale may be used as an index for measuring efficacy of an antibody. The Sartorius scale is described by Sartorius et al. in British Journal of Dermatology, 149: 211-213. Briefly, the following outcome variables are explicitly mentioned in reports based on the Sartorius scale: (1) anatomical region involved (axilla, groin, gluteal or other region or inframammary region left and / or right: 3 points per region involved); (2) number and scores of lesions (abscesses, nodules, fistulas, scars: points per lesion of all regions involved: nodules 2; fistulas 4; scars 1; others 1); (3) the longest distance between two relevant lesions, i.e., nodules and fistulas, in each region, or size if only one lesion (< 5 cm, 2; < 10 cm, 4; > 10 cm, 8); and (4) are all lesions clearly separated by normal skin. In each region (yes 0 / no 6). By assigning numerical scores to these variables, disease intensity can be quantified in a more clinically meaningful way on an open-ended scale. A total score as well as scores of selected regions chosen for surgical or other intervention can be calculated and followed over time.

[0278] In some examples, treatment of HS with a compound disclosed herein is determined according to achieving an HiSCR (Hidradenitis Suppurativa Clinical Response) of the subject being treated. The HisSCR is defined as at least a 50% reduction in the total inflammatory lesion (abscess and inflammatory nodule) count (AN count) in a subject relative to baseline, with no increase in abscess count and no increase in draining fistula count. In one instance, treatment of HS in a subject is defined as an at least 50% reduction in the inflammatory lesion (abscess and nodule) count. The HiSCR scoring system was designed to assess hidradenitis suppurativa activity in an affected subject before and after a treatment.

[0279] In another example, treatment of HS with a compound disclosed herein is defined as achieving an Physician's Global Assessment (PGA) score as defined in Table 2 below, of clear (0), minimal (1), or mild (2), with an improvement (i.e., reduction) from baseline PGA score of at least 1 grade or 2 grades, optionally, at the end of a treatment period (such as week 16). The baseline PGA score is the PGA score measured just prior to the commencement of treatment, to which the PGA score obtained after a period of treatment is compared.

[0280] Table 2. HS PGA Scoring.

[0281] In one example, the present disclosure provides a method for improving the DLQI score of a subject suffering from HS. In one instance, the improvement in the DLQI score is determined by achieving a score, e.g., a statistically significant score, correlating with a "no" or "small impact" of the disease state on the subject. In one example, the present disclosure provides a method for decreasing the number of inflammatory lesions (AN count) in a subject having HS, said method comprising administering an antibody disclosed herein to the subject, such that the AN count is decreased. The decrease in AN count may be anything greater than 10%, e.g., the AN count may be reduced by at least a 50% reduction in the subject relative to baseline AN count. The subject may also exhibit other improvements in HS following treatment with an antibody disclosed herein, for example the subject may have no increase in an abscess count and / or no increase in a draining fistula count following administration with the antibody.

[0282] In one example, the neutrophil-associated skin condition is palmoplantar pustulosis (PPP). PPP is a chronic pustular condition affecting the hands and / or soles of the feet. PPP can occur with psoriasis or without any skin disease. PPP affects the eccrine sweat glands which are most common on the palms and soles. PPP presents as crops of itchy or sore pustules on the palms and / or soles. PPP can occur on one or both hands and / or feet. Scaly red patches may also be seen in association with the pustules. In the more chronic stages of the disease, the skin can be dry and thickened with deep fissures (cracks in the skin). There is usually a sharp demarcation between the normal and affected skin areas. PPP varies in severity and may persist for many years. The discomfort can be considerable, interfering with work and affecting quality of life. A form of PPP which affects the tips of the fingers is called acrodermatitis continua of Hallopeau or acropustulosis. It can lead to destruction of the fingernail on affected digits.

[0283] Certain subtypes of PPP may be treated in accordance with the present disclosure. In one instance, moderate to severe PPP, is treated by administering an antibody disclosed herein. In one example, chronic PPP, e.g., moderate to severe chronic PPP, is treated by administering an antibody disclosed herein. In one example, the subject has confirmed moderate to severe PPP, e.g., confirmed chronic moderate to severe PPP.

[0284] The present disclosure also provides a method for treating certain subpopulations of PPP patients who may be especially difficult to treat, i.e., patients who have a subtherapeutic response to an anti-TNF therapy for their PPP.

[0285] The present disclosure also provides methods for improving PPP symptoms in a subject based on indices used to measure the disease state. Severity of disease and / or treatment of PPP using a compound disclosed herein may be determined using measures known in the art. e.g., improvement in ppPASI, or any measure known to those in the art.

[0286] The ppPASI is an assessment tool based on the Psoriasis Area and Severity Index that is widely used for assessing severity of chronic plaque psoriasis. Parameters including severity, erythema, total number of pustules and desquamation are scored on a scale of 1-4, then corrected for area and site involved (palm or sole). The sum of the four values produces the final ppPASI which ranges between 0 (no PPP) and 72 (the most severe PPP) (Bhushan, et al., Br J Dermatol, 145: 546-53, 2001). ppPASI can be assessed at screening, prior to administration. In one example, administration of a compound as disclosed herein reduces a ppPASI score. In one example, a subject has a ppPASI score of >12 prior to commencing treatment as described herein. In one example, a subject has a ppPASI score of <12 following treatment as described herein.

[0287] In one example, administration of a compound as disclosed herein reduces Palm- Sole Physician Global Assessment (PGA) score in a subject suffering from PPP. The PGA is an average assessment of all psoriatic lesions based on erythema, scale, and induration (Robinson, 2011). PGA can be assessed prior to administration of the antibody.

[0288] Other response indices for PPP include: the PASI scoring system, Investigator's Global Assessment mod 2011 (IGA mod 2011), Dermatology Life Quality Index (DLQI) and Subject's Global Assessment (SGA), Work Productivity and Activity Impairment Questionnaire-Psoriasis (WPALPSO), Palmar-Pustular Quality of Life Index (ppQoL- Index).

[0289] In one example, the neutrophil-associated condition is psoriasis. As used herein, the term “psoriasis” encompasses all subtypes of psoriasis, including plaque, guttate, inverse, pustular, and erythrodermic. In one example, the neutrophil-mediated condition is plaque psoriasis (also known in the art as “psoriasis vulgaris” or “common psoriasis”). Certain subtypes of psoriasis may be treated in accordance with the present disclosure. In one instance, moderate to severe psoriasis, is treated by administering a compound disclosed herein. In one example, the subject has confirmed moderate to severe psoriasis, e.g., chronic moderate to severe psoriasis.

[0290] The present disclosure also provides a method for treating certain subpopulations of psoriasis patients who may be especially difficult to treat, i.e., patients who have been unresponsive or intolerant to an anti-TNF therapy for treatment for their psoriasis.

[0291] The present disclosure also provides methods for improving psoriasis symptoms in a subject based on indices used to measure the disease state. Treatment of psoriasis using a compound disclosed herein may also be determined using measures known in the art.

[0292] Methods for measuring the severity of psoriasis and / or response to treatment will be apparent to the skilled artisan. For example, the Psoriasis Area and Severity Index (PASI) is used by dermatologists to assess psoriasis disease intensity. This index is based on the quantitative assessment of three typical signs of psoriatic lesions: erythema, infiltration, and desquamation, combined with the skin surface area involvement. Since its development in 1978, this instrument has been used throughout the world by clinical investigators (Fredriksson T, Petersson U: Dermatologica 1978; 157: 238-41). PASI is indicated as PASI 50 (a 50 percent improvement in PASI from baseline), PASI 75 (a 75 percent improvement in PASI from baseline), PASI 90 (a 90 percent improvement in PASI from baseline), and PASI 100 (a 100 percent improvement in PASI from baseline).

[0293] The Physicians Global Assessment (PGA) is used to assess psoriasis activity and follow clinical response to treatment. It is a six-point score that summarizes the overall quality (erythema, scaling and thickness) and extent of plaques relative to the baseline assessment. A patient's response is rated as worse, poor (0-24%), fair (25-49%), good (50-74%), excellent (75-99%), or cleared (100%) (van der Kerkhof P. Br J Dermatol 737:661-662, 1997). Other measures of improvements in the disease state of a subject having psoriasis include clinical responses, such as the Dermatology Life Quality Index (DLQI) and the Minimum Clinically Important Difference (MCID).

[0294] Antibodies

[0295] In one example, a compound as described herein according to any example is a protein comprising an antigen binding site of an antibody. In some examples, the compound that inhibits G-CSF signaling is an antibody. In some examples, the antibody binds to G-CSFR. In some examples, the antibody binds to G-CSF.

[0296] Methods for generating antibodies are known in the art and / or described in Harlow and Lane (editors) Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, (1988). Generally, in such methods G-CSFR or G-CSF (e.g., hG-CSFR or hG-CSF) or a region thereof (e.g., an extracellular domain) or immunogenic fragment or epitope thereof or a cell expressing and displaying same (i.e., an immunogen), optionally formulated with any suitable or desired carrier, adjuvant, or pharmaceutically acceptable excipient, is administered to a non-human animal, for example, a mouse, chicken, rat, rabbit, guinea pig, dog, horse, cow, goat or pig. The immunogen may be administered intranasally, intramuscularly, sub-cutaneously, intravenously, intradermally, intraperitoneally, or by other known route.

[0297] Monoclonal antibodies are one exemplary form of an antibody contemplated by the present disclosure. The term “monoclonal antibody" or “mAb” refers to a homogeneous antibody population capable of binding to the same antigen(s), for example, to the same epitope within the antigen. This term is not intended to be limited as regards to the source of the antibody or the manner in which it is made. For the production of mAbs any one of a number of known techniques may be used, such as, for example, the procedure exemplified in US4196265 or Harlow and Lane (1988), supra.

[0298] Alternatively, ABL-MYC technology (NeoClone, Madison WI 53713, USA) is used to produce cell lines secreting MAbs (e.g., as described in Largaespada et al, J. Immunol. Methods. 197 85-95, 1996).

[0299] Antibodies can also be produced or isolated by screening a display library, e.g., a phage display library, e.g., as described in US6300064 and / or US5885793. For example, the present inventors have isolated fully human antibodies from a phage display library.

[0300] The antibody of the present disclosure may be a synthetic antibody. For example, the antibody is a chimeric antibody, a humanized antibody, a human antibody or a deimmunized antibody.

[0301] In one example, an antibody described herein is a chimeric antibody. The term “chimeric antibody” refers to antibodies in which a portion of the heavy and / or light chain is identical with or homologous to corresponding sequences in antibodies derived from a particular species (e.g., murine, such as mouse) or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical with or homologous to corresponding sequences in antibodies derived from another species (e.g., primate, such as human) or belonging to another antibody class or subclass. Methods for producing chimeric antibodies are described in, e.g., US4816567; and US5807715.

[0302] The antibodies of the present disclosure may be humanized or human.

[0303] The term "humanized antibody” shall be understood to refer to a subclass of chimeric antibodies having an antigen binding site or variable region derived from an antibody from a non-human species and the remaining antibody structure based upon the structure and / or sequence of a human antibody. In a humanized antibody, the antigenbinding site generally comprises the complementarity determining regions (CDRs) from the non-human antibody grafted onto appropriate FRs in the variable regions of a human antibody and the remaining regions from a human antibody. Antigen binding sites may be wild-type (i.e., identical to those of the non-human antibody) or modified by one or more amino acid substitutions. In some instances, FR residues of the human antibody are replaced by corresponding non-human residues.

[0304] Methods for humanizing non-human antibodies or parts thereof (e.g., variable regions) are known in the art. Humanization can be performed following the method of US5225539, or US5585089. Other methods for humanizing an antibody are not excluded. The term "human antibody" as used herein refers to antibodies having variable regions (e.g. VH, VL) and, optionally constant regions derived from or corresponding to sequences found in humans, e.g. in the human germline or somatic cells.

[0305] Exemplary human antibodies are described herein and include Cl.2 and C1.2G and / or variable regions thereof. These human antibodies provide an advantage of reduced immunogenicity in a human compared to non-human antibodies. Exemplary antibodies that bind to G-CSFR and inhibit G-CSF signaling are described in W02012 / 171057, for example. In some examples, the antibody binds to G-CSF. Exemplary antibodies that bind to G-CSF and inhibit G-CSF signaling are described in WO2018 / 145206, for example. One particular exemplary antibody is anumigilimab (also referred to herein as “CSE324”).

[0306] Antibody Binding Domain Containing Proteins

[0307] Single-Domain Antibodies

[0308] In some examples, a compound of the disclosure is a protein that is or comprises a single-domain antibody (which is used interchangeably with the term “domain antibody” or “dAb”). A single-domain antibody is a single polypeptide chain comprising all or a portion of the heavy chain variable region of an antibody. In certain examples, a single-domain antibody is a human single-domain antibody (Domantis, Inc., Waltham, MA; see, e.g., US6248516).

[0309] Diabodies, Triabodies, Tetrabodies

[0310] In some examples, a protein of the disclosure is or comprises a diabody, triabody, tetrabody or higher order protein complex such as those described in W098 / 044001 and / or WG94 / 007921.

[0311] Single Chain Fv (scFv)

[0312] The skilled artisan will be aware that scFvs comprise VH and VL regions in a single polypeptide chain and a polypeptide linker between the VH and VL which enables the scFv to form the desired structure for antigen binding (i.e., for the VH and VL of the single polypeptide chain to associate with one another to form a Fv). For example, the linker comprises in excess of 12 amino acid residues with (Gly4Ser)3 being one of the more favored linkers for a scFv.

[0313] Heavy Chain Antibodies Heavy chain antibodies differ structurally from many other forms of antibodies, in so far as they comprise a heavy chain, but do not comprise a light chain. Accordingly, these antibodies are also referred to as “heavy chain only antibodies”. Heavy chain antibodies are found in, for example, camelids and cartilaginous fish (also called IgNAR).

[0314] A general description of heavy chain antibodies from camelids and the variable regions thereof and methods for their production and / or isolation and / or use is found inter alia in the following references WO94 / 04678, WO97 / 49805 and WO 97 / 49805.

[0315] A general description of heavy chain antibodies from cartilaginous fish and the variable regions thereof and methods for their production and / or isolation and / or use is found inter alia in W02005 / 118629.

[0316] Other Antibodies and Antibody Fragments

[0317] The present disclosure also contemplates other antibodies and antibody fragments, such as:

[0318] (i) “key and hole” bispecific proteins as described in US5, 731,168;

[0319] (ii) heteroconjugate proteins, e.g., as described in US4,676,980;

[0320] (iii) heteroconjugate proteins produced using a chemical cross-linker, e.g., as described in US4,676,980; and

[0321] (iv) Fabs (e.g., as described in EP19930302894).

[0322] V-Like Proteins

[0323] An example of a compound of the disclosure is a T-cell receptor. T cell receptors have two V-domains that combine into a structure similar to the Fv module of an antibody. Novotny et al., Proc Natl Acad Sci USA 88 8646-8650, 1991 describes how the two V-domains of the T-cell receptor (termed alpha and beta) can be fused and expressed as a single chain polypeptide and, further, how to alter surface residues to reduce the hydrophobicity directly analogous to an antibody scFv. Other publications describing production of single-chain T-cell receptors or multimeric T cell receptors comprising two V-alpha and V-beta domains include W01999 / 045110 or WO201 1 / 107595.

[0324] Other non-antibody proteins comprising antigen binding domains include proteins with V-like domains, which are generally monomeric. Examples of proteins comprising such V-like domains include CTLA-4, CD28 and ICOS. Further disclosure of proteins comprising such V-like domains is included in WO 1999 / 045110. Adnectins

[0325] In one example, a compound of the disclosure is an adnectin. Adnectins are based on the tenth fibronectin type III (10Fn3) domain of human fibronectin in which the loop regions are altered to confer antigen binding. For example, three loops at one end of the P-sandwich of the10Fn3 domain can be engineered to enable an Adnectin to specifically recognize an antigen. For further details see US20080139791 or W02005 / 056764.

[0326] Anticalins

[0327] In a further example, a compound of the disclosure is an anticalin. Anticalins are derived from lipocalins, which are a family of extracellular proteins which transport small hydrophobic molecules such as steroids, bilins, retinoids and lipids. Lipocalins have a rigid P-sheet secondary structure with a plurality of loops at the open end of the conical structure which can be engineered to bind to an antigen. Such engineered lipocalins are known as anticalins. For further description of anticalins see US7250297B1 or US20070224633.

[0328] Affibodies

[0329] In a further example, a compound of the disclosure is an affibody. An affibody is a scaffold derived from the Z domain (antigen binding domain) of Protein A of Staphylococcus aureus which can be engineered to bind to antigen. The Z domain consists of a three -helical bundle of approximately 58 amino acids. Libraries have been generated by randomization of surface residues. For further details see EP1641818.

[0330] Avimers

[0331] In a further example, a compound of the disclosure is an Avimer. Avimers are multidomain proteins derived from the A-domain scaffold family. The native domains of approximately 35 amino acids adopt a defined disulfide bonded structure. Diversity is generated by shuffling of the natural variation exhibited by the family of A-domains. For further details see W02002088171.

[0332] DARPins

[0333] In a further example, a compound of the disclosure is a Designed Ankyrin Repeat Protein (DARPin). DARPins are derived from Ankyrin which is a family of proteins that mediate attachment of integral membrane proteins to the cytoskeleton. A single ankyrin repeat is a 33 residue motif consisting of two a-helices and a P-turn. They can be engineered to bind different target antigens by randomizing residues in the first a-helix and a P-turn of each repeat. Their binding interface can be increased by increasing the number of modules (a method of affinity maturation). For further details see US20040132028.

[0334] Soluble G-CSFR

[0335] The present disclosure also contemplates a soluble form of the G-CSFR which competes with the naturally occurring membrane-associated G-CSFR for G-CSF interaction. Those skilled in the art can readily prepare soluble forms of the receptor, see for example U.S. Pat. No. 5,589,456 and Honjo et al, Acta Cry stallo graph Sect F Struct Biol Cryst Commun. 61(Pt 8):788-790, 2005.

[0336] De-immunized Antibodies and Proteins

[0337] The present disclosure also contemplates a de-immunized antibody or protein. De-immunized antibodies and proteins have one or more epitopes, e.g., B cell epitopes or T cell epitopes removed (i.e., mutated) to thereby reduce the likelihood that a mammal will raise an immune response against the antibody or protein. Methods for producing de-immunized antibodies and proteins are known in the art and described, for example, in WG2000 / 34317, WG2004 / 108158 and WG2004 / 064724.

[0338] Methods for introducing suitable mutations and expressing and assaying the resulting protein will be apparent to the skilled artisan based on the description herein.

[0339] Mutations to Proteins

[0340] The present disclosure also contemplates mutant forms of a protein of the disclosure. In this regard, data presented herein indicate sites within a CDR of a protein of the disclosure that can be changed in addition to exemplary changes that can be made. The skilled person will understand that changes can additionally or alternatively be made within a framework region of a variable region containing protein without inhibiting or significantly reducing its function in the context of the present disclosure.

[0341] For example, such a mutant protein comprises one or more conservative amino acid substitutions compared to a sequence set forth herein. In some examples, the protein comprises 30 or fewer or 20 or fewer or 10 or fewer, e.g., 9 or 8 or 7 or 6 or 5 or 4 or 3 or 2 conservative amino acid substitutions. A "conservative amino acid substitution" is one in which the amino acid residue is replaced with an amino acid residue having a similar side chain and / or hydropathicity and / or hydrophilicity.

[0342] In one example, a mutant protein has only, or not more than, one or two or three or four or five or six conservative amino acid changes when compared to a naturally occurring protein. Details of conservative amino acid changes are provided below. As the skilled person would be aware, e.g., from the disclosure herein, such minor changes can reasonably be predicted not to alter the activity of the protein.

[0343] Families of amino acid residues having similar side chains have been defined in the art, including basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), P-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine).

[0344] The present disclosure also contemplates non-conservative amino acid changes (e.g., substitutions) in a protein of the present disclosure, e.g., in a CDR, such as CDR3. For example, the present inventors have identified several non-conservative amino acid substitutions that can be made while retaining an activity of a protein of the disclosure. In one example, the protein comprises fewer than 6 or 5 or 4 or 3 or 2 or 1 non- conservative amino acid substitutions, e.g., in a CDR3, such as in a CDR3.

[0345] The present disclosure also contemplates one or more insertions or deletions compared to a sequence set forth herein. In some examples, the protein comprises 10 or fewer, e.g., 9 or 8 or 7 or 6 or 5 or 4 or 3 or 2 insertions and / or deletions.

[0346] Constant Regions

[0347] The present disclosure encompasses proteins and / or antibodies described herein comprising a constant region of an antibody. This includes antigen binding fragments of an antibody fused to a Fc.

[0348] Sequences of constant regions useful for producing the proteins of the present disclosure may be obtained from a number of different sources. In some examples, the constant region or portion thereof of the protein is derived from a human antibody. The constant region or portion thereof may be derived from any antibody class, including IgM, IgG, IgD, IgA and IgE, and any antibody isotype, including IgGl, IgG2, IgG3 and IgG4. In one example, the constant region is human isotype IgG4 or a stabilized IgG4 constant region.

[0349] In one example, the Fc region of the constant region has a reduced ability to induce effector function, e.g., compared to a native or wild-type human IgGl or IgG3 Fc region. In one example, the effector function is antibody -dependent cell-mediated cytotoxicity (ADCC) and / or antibody -dependent cell-mediated phagocytosis (ADCP) and / or complement-dependent cytotoxicity (CDC). Methods for assessing the level of effector function of an Fc region containing protein are known in the art and / or described herein.

[0350] In one example, the Fc region is an IgG4 Fc region (i.e., from an IgG4 constant region), e.g., a human IgG4 Fc region. Sequences of suitable IgG4 Fc regions will be apparent to the skilled person and / or available in publically available databases (e.g., available from National Center for Biotechnology Information).

[0351] In one example, the constant region is a stabilized IgG4 constant region. The term “stabilized IgG4 constant region” will be understood to mean an IgG4 constant region that has been modified to reduce Fab arm exchange or the propensity to undergo Fab arm exchange or formation of a half-antibody or a propensity to form a half antibody. “Fab arm exchange" refers to a type of protein modification for human IgG4, in which an IgG4 heavy chain and attached light chain (half-molecule) is swapped for a heavy-light chain pair from another IgG4 molecule. Thus, IgG4 molecules may acquire two distinct Fab arms recognizing two distinct antigens (resulting in bispecific molecules). Fab arm exchange occurs naturally in vivo and can be induced in vitro by purified blood cells or reducing agents such as reduced glutathione. A “half antibody” forms when an IgG4 antibody dissociates to form two molecules each containing a single heavy chain and a single light chain.

[0352] In one example, a stabilized IgG4 constant region comprises a proline at position 241 of the hinge region according to the system of Kabat (Kabat et al., Sequences of Proteins of Immunological Interest Washington DC United States Department of Health and Human Services, 1987 and / or 1991). This position corresponds to position 228 of the hinge region according to the EU numbering system (Kabat et al., Sequences of Proteins of Immunological Interest Washington DC United States Department of Health and Human Services, 2001 and Edelman et al., Proc. Natl. Acad. USA, 63, 78-85, 1969). In human IgG4, this residue is generally a serine. Following substitution of the serine for proline, the IgG4 hinge region comprises a sequence CPPC. In this regard, the skilled person will be aware that the “hinge region” is a proline-rich portion of an antibody heavy chain constant region that links the Fc and Fab regions that confers mobility on the two Fab arms of an antibody. The hinge region includes cysteine residues which are involved in inter-heavy chain disulfide bonds. It is generally defined as stretching from Glu226 to Pro243 of human IgGl according to the numbering system of Kabat. Hinge regions of other IgG isotypes may be aligned with the IgGl sequence by placing the first and last cysteine residues forming inter-heavy chain disulphide (S-S) bonds in the same positions (see for example W02010 / 080538). Additional examples of stabilized IgG4 antibodies are antibodies in which arginine at position 409 in a heavy chain constant region of human IgG4 (according to the EU numbering system) is substituted with lysine, threonine, methionine, or leucine (e.g., as described in W02006 / 033386). The Fc region of the constant region may additionally or alternatively comprise a residue selected from the group consisting of: alanine, valine, glycine, isoleucine and leucine at the position corresponding to 405 (according to the EU numbering system). Optionally, the hinge region comprises a proline at position 241 (i.e., a CPPC sequence) (as described above).

[0353] In another example, the Fc region is a region modified to have reduced effector function, i.e., a “non-immunostimulatory Fc region”. For example, the Fc region is an IgGl Fc region comprising a substitution at one or more positions selected from the group consisting of 268, 309, 330 and 331. In another example, the Fc region is an IgGl Fc region comprising one or more of the following changes E233P, E234V, E235A and deletion of G236 and / or one or more of the following changes A327G, A33OS and P33 IS (Armour et al., Eur J Immunol. 29:2613-2624, 1999; Shields et al., J Biol Chem. 276(9):6591-604, 2001). Additional examples of non-immunostimulatory Fc regions are described, for example, in Dall'Acqua et al., J Immunol. 177 : 1129-1138 2006; and / or Hezareh J Virol ;75 12161-12168, 2001).

[0354] In another example, the Fc region is a chimeric Fc region, e.g., comprising at least one CH2 domain from an IgG4 antibody and at least one CH3 domain from an IgGl antibody, wherein the Fc region comprises a substitution at one or more amino acid positions selected from the group consisting of 240, 262, 264, 266, 297, 299, 307, 309, 323, 399, 409 and 427 (EU numbering) (e.g., as described in W02010 / 085682). Exemplary substitutions include 240F, 262E, 264T, 266F, 297Q, 299A, 299K, 307P, 309K, 309M, 309P, 323F, 399S, and 427F.

[0355] Additional Modifications / Variants

[0356] The present disclosure also contemplates additional modifications or variants to an antibody or protein of the disclosure.

[0357] For example, the antibody comprises one or more amino acid substitutions that increase the half-life of the protein. For example, the antibody comprises a Fc region comprising one or more amino acid substitutions that increase the affinity of the Fc region for the neonatal Fc region (FcRn). For example, the Fc region has increased affinity for FcRn at lower pH, e.g., about pH 6.0, to facilitate Fc / FcRn binding in an endosome. In one example, the Fc region has increased affinity for FcRn at about pH 6 compared to its affinity at about pH 7.4, which facilitates the re-release of Fc into blood following cellular recycling. These amino acid substitutions are useful for extending the half life of a protein, by reducing clearance from the blood.

[0358] Exemplary amino acid substitutions include T250Q and / or M428L or T252A, T254S and T266F or M252Y, S254T and T256E or H433K and N434F according to the EU numbering system. Additional or alternative amino acid substitutions are described, for example, in US20070135620 or US7083784.

[0359] In one example, the protein of the disclosure additionally comprises albumin, a functional fragment or variant thereof. In one example, the albumin, functional fragment or variant thereof is serum albumin, such as human serum albumin. In one example, the albumin, functional fragment or variant thereof, comprises one or more amino acid substitutions, deletions or insertions, e.g., no more than 5 or 4 or 3 or 2 or 1 substitutions. Amino acid substitutions suitable for use in the present disclosure will be apparent to the skilled person and include naturally-occurring substitutions and engineered substitutions such as those described, for example, in WO2011051489, WO2014072481, WO201 1103076, WO2012112188, W02013075066, W02015063611 and

[0360] WO2014179657.

[0361] In one example, the protein or antibody of the disclosure comprises one or more variants. For example, the variant is a post-translationally modified variant.

[0362] In one example, the protein or antibody comprises a variant missing an encoded C-terminal lysine residue, a deamidated variant, a miscorporated amino acid residue, a glycosylated variant, a variant comprising a pyroglutamate, a variant lacking a N- terminal residue, and / or a variant comprising all or part of a secretion signal.

[0363] In one example, the protein or antibody comprises a variant missing an encoded C-terminal lysine residue.

[0364] In one example, the protein or antibody comprises a deamidated variant. Deamidated variants of encoded asparagine residues may result in isoaspartic acid and / or aspartic acid being generated or even a succinamide involving an adjacent amino acid residue. Deamidated variants of encoded glutamine residues may result in glutamic acid being formed. Compositions comprising a heterogeneous mixture of such sequences and variants are intended to be included when reference is made to a particular amino acid sequence.

[0365] In one example, the protein or antibody comprises a miscorporated amino acid residue. For example, methionine residues are substituted with nor-leucine, asparagine residues are substituted with serine and / or phenylalanine residues are substituted with tyrosine. In one example, the protein or antibody comprises a variant comprising a pyroglutamate. For example, at the N-terminus of a protein.

[0366] In one example, the protein or antibody comprises a glycosylated variant.

[0367] In one example, the protein or antibody comprises a variant lacking a N-terminal residue. For example, a N-terminal glutamine in an antibody or V region.

[0368] In one example, the protein or antibody comprises a variant comprising all or part of a secretion signal. As the skilled person will understand, a population of antibodies in a composition may include antibodies without one of the aforementioned variants and other antibodies with one or more of the aforementioned variants (i.e., a mixture of variant and non- variant antibodies).

[0369] Protein Production

[0370] In one example, a protein described herein according to any example is produced by culturing a hybridoma under conditions sufficient to produce the protein, e.g., as described herein and / or as is known in the art and / or may be produced via recombinant expression.

[0371] Recombinant Expression

[0372] In another example, a protein described herein according to any example is recombinant.

[0373] In the case of a recombinant protein, nucleic acid encoding same can be cloned into expression constructs or vectors, which are then transfected into host cells, such as E. coli cells, yeast cells, insect cells, or mammalian cells, such as simian COS cells, Chinese Hamster Ovary (CHO) cells, human embryonic kidney (HEK) cells, or myeloma cells that do not otherwise produce the protein. Exemplary cells used for expressing a protein are CHO cells, myeloma cells or HEK cells. Molecular cloning techniques to achieve these ends are known in the art and described, for example in Ausubel et al., (editors), Current Protocols in Molecular Biology, Greene Pub. Associates and Wiley- Interscience (1988, including all updates until present) or Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press (1989). A wide variety of cloning and in vitro amplification methods are suitable for the construction of recombinant nucleic acids. Methods of producing recombinant antibodies are also known in the art, see, e.g., US4816567 or US5530101.

[0374] Following isolation, the nucleic acid is inserted operably linked to a promoter in an expression construct or expression vector for further cloning (amplification of the DNA) or for expression in a cell-free system or in cells. As used herein, the term “promoter” is to be taken in its broadest context and includes the transcriptional regulatory sequences of a genomic gene, including the TATA box or initiator element, which is required for accurate transcription initiation, with or without additional regulatory elements (e.g., upstream activating sequences, transcription factor binding sites, enhancers and silencers) that alter expression of a nucleic acid, e.g., in response to a developmental and / or external stimulus, or in a tissue specific manner. In the present context, the term “promoter” is also used to describe a recombinant, synthetic or fusion nucleic acid, or derivative which confers, activates or enhances the expression of a nucleic acid to which it is operably linked. Exemplary promoters can contain additional copies of one or more specific regulatory elements to further enhance expression and / or alter the spatial expression and / or temporal expression of said nucleic acid.

[0375] As used herein, the term “operably linked to" means positioning a promoter relative to a nucleic acid such that expression of the nucleic acid is controlled by the promoter.

[0376] Many vectors for expression in cells are available. The vector components generally include, but are not limited to, one or more of the following: a signal sequence, a sequence encoding a protein (e.g., derived from the information provided herein), an enhancer element, a promoter, and a transcription termination sequence. The skilled artisan will be aware of suitable sequences for expression of a protein. Exemplary signal sequences include prokaryotic secretion signals (e.g., pelB, alkaline phosphatase, penicillinase, Ipp, or heat-stable enterotoxin II), yeast secretion signals (e.g., invertase leader, a factor leader, or acid phosphatase leader) or mammalian secretion signals (e.g., herpes simplex gD signal).

[0377] Exemplary promoters active in mammalian cells include cytomegalovirus immediate early promoter (CMV-IE), human elongation factor 1-a promoter (EFl), small nuclear RNA promoters (Ula and Ulb), a-myosin heavy chain promoter, Simian virus 40 promoter (SV40), Rous sarcoma virus promoter (RSV), Adenovirus major late promoter, P-actin promoter; hybrid regulatory element comprising a CMV enhancer / P- actin promoter or an immunoglobulin promoter or active fragment thereof. Examples of useful mammalian host cell lines are monkey kidney CV1 line transformed by SV40 (COS-7, ATCC CRL 1651); human embryonic kidney line (293 or 293 cells subcloned for growth in suspension culture; baby hamster kidney cells (BHK, ATCC CCL 10); or Chinese hamster ovary cells (CHO).

[0378] Typical promoters suitable for expression in yeast cells such as for example a yeast cell selected from the group comprising Pichia pastoris, Saccharomyces cerevisiae and S. pombe, include, but are not limited to, the ADH1 promoter, the GALI promoter, the GAIA promoter, the CUP1 promoter, the PHO 5 promoter, the nmt promoter, the RPR1 promoter, or the TEF1 promoter.

[0379] Means for introducing the isolated nucleic acid or expression construct comprising same into a cell for expression are known to those skilled in the art. The technique used for a given cell depends on the known successful techniques. Means for introducing recombinant DNA into cells include microinjection, transfection mediated by DEAE-dextran, transfection mediated by liposomes such as by using lipofectamine (Gibco, MD, USA) and / or cellfectin (Gibco, MD, USA), PEG-mediated DNA uptake, electroporation and microparticle bombardment such as by using DNA-coated tungsten or gold particles (Agracetus Inc., WI, USA) amongst others.

[0380] The host cells used to produce the protein may be cultured in a variety of media, depending on the cell type used. Commercially available media such as Ham's F10 (Sigma), Minimal Essential Medium ((MEM), (Sigma), RPM1-1640 (Sigma), and Dulbecco's Modified Eagle's Medium ((DMEM), Sigma) are suitable for culturing mammalian cells. Media for culturing other cell types discussed herein are known in the art.

[0381] Isolation of Proteins

[0382] Methods for isolating a protein are known in the art and / or described herein.

[0383] Where a protein is secreted into culture medium, supernatants from such expression systems can be first concentrated using a commercially available protein concentration filter, for example, an Amicon or Millipore Pellicon ultrafiltration unit. A protease inhibitor such as PMSF may be included in any of the foregoing steps to inhibit proteolysis and antibiotics may be included to prevent the growth of adventitious contaminants. Alternatively, or additionally, supernatants can be filtered and / or separated from cells expressing the protein, e.g., using continuous centrifugation.

[0384] The protein prepared from the cells can be purified using, for example, ion exchange, hydroxyapatite chromatography, hydrophobic interaction chromatography, gel electrophoresis, dialysis, affinity chromatography (e.g., protein A affinity chromatography or protein G chromatography), or any combination of the foregoing. These methods are known in the art and described, for example in WO99 / 57134 or Ed Harlow and David Lane (editors) Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, (1988).

[0385] The skilled artisan will also be aware that a protein can be modified to include a tag to facilitate purification or detection, e.g., a poly -histidine tag, e.g., a hexa-histidine tag, or an influenza virus hemagglutinin (HA) tag, or a Simian Virus 5 (V5) tag, or a FLAG tag, or a glutathione S-transferase (GST) tag. The resulting protein is then purified using methods known in the art, such as, affinity purification. For example, a protein comprising a hexa-his tag is purified by contacting a sample comprising the protein with nickel-nitrilotriacetic acid (Ni-NTA) that specifically binds a hexa-his tag immobilized on a solid or semi-solid support, washing the sample to remove unbound protein, and subsequently eluting the bound protein. Alternatively, or in addition a ligand or antibody that binds to a tag is used in an affinity purification method.

[0386] Nucleic Acid-Based G-CSF Signaling Inhibitors

[0387] In one example of the disclosure, therapeutic and / or prophylactic methods as described herein according to any example of the disclosure involve reducing expression of G-CSF and / or G-CSFR. For example, such a method involves administering a compound that reduces transcription and / or translation of a nucleic acid encoding G-CSF or G-CSFR. In one example, the compound that inhibits G-CSF signaling is a nucleic acid, e.g., an antisense polynucleotide, a ribozyme, a PNA, an interfering RNA, a siRNA, a microRNA.

[0388] In another example, the compound that inhibits G-CSF signaling is a nucleic acid encoding a protein compound that inhibits G-CSF signaling (e.g., an antibody or antigen binding fragment thereof).

[0389] Antisense Nucleic Acids

[0390] The term “antisense nucleic acid” shall be taken to mean a DNA or RNA or derivative thereof (e.g., LNA or PNA), or combination thereof that is complementary to at least a portion of a specific mRNA molecule encoding a polypeptide as described herein in any example of the disclosure and capable of interfering with a post- transcriptional event such as mRNA translation. The use of antisense methods is known in the art (see for example, Hartmann and Endres (editors), Manual of Antisense Methodology, Kluwer (1999)).

[0391] An antisense nucleic acid of the disclosure will hybridize to a target nucleic acid under physiological conditions. Antisense nucleic acids include sequences that correspond to structural genes or coding regions or to sequences that effect control over gene expression or splicing. For example, the antisense nucleic acid may correspond to the targeted coding region of a nucleic acid encoding G-CSF or G-CSFR, or the 5’- untranslated region (UTR) or the 3’-UTR or combination of these. It may be complementary in part to intron sequences, which may be spliced out during or after transcription, for example only to exon sequences of the target gene. The length of the antisense sequence should be at least 19 contiguous nucleotides, for example, at least 50 nucleotides, such as at least 100, 200, 500 or 1000 nucleotides of a nucleic acid encoding G-CSF or G-CSFR. The full-length sequence complementary to the entire gene transcript may be used. The length can be 100-2000 nucleotides. The degree of identity of the antisense sequence to the targeted transcript should be at least 90%, for example, 95- 100%.

[0392] Exemplary antisense nucleic acids against G-CSF or G-CSFR are described, for example, in WO2011032204.

[0393] Catalytic Nucleic Acid

[0394] The term “catalytic nucleic acid” refers to a DNA molecule or DNA-containing molecule (also known in the art as a “deoxyribozyme” or “DNAzyme”) or a RNA or RNA-containing molecule (also known as a “ribozyme” or “RNAzyme”) which specifically recognizes a distinct substrate and catalyzes the chemical modification of this substrate. The nucleic acid bases in the catalytic nucleic acid can be bases A, C, G, T (and U for RNA).

[0395] Typically, the catalytic nucleic acid contains an antisense sequence for specific recognition of a target nucleic acid, and a nucleic acid cleaving enzymatic activity (also referred to herein as the “catalytic domain”). The types of ribozymes that are useful in this disclosure are a hammerhead ribozyme and a hairpin ribozyme.

[0396] RNA Interference

[0397] RNA interference (RNAi) is useful for specifically inhibiting the production of a particular protein. Without being limited by theory, this technology relies on the presence of dsRNA molecules that contain a sequence that is essentially identical to the mRNA of the gene of interest or part thereof, in this case an mRNA encoding G-CSF or G-CSFR. Conveniently, the dsRNA can be produced from a single promoter in a recombinant vector host cell, where the sense and anti-sense sequences are flanked by an unrelated sequence which enables the sense and anti-sense sequences to hybridize to form the dsRNA molecule with the unrelated sequence forming a loop structure. The design and production of suitable dsRNA molecules for the present disclosure is well within the capacity of a person skilled in the art, particularly considering WO99 / 32619, WG99 / 53050, WO99 / 49029, and WO01 / 34815. Such dsRNA molecules for RNAi include, but are not limited to short hairpin RNA (shRNA) and bi-functional shRNA. The length of the sense and antisense sequences that hybridize should each be at least 19 contiguous nucleotides, such as at least 30 or 50 nucleotides, for example at least 100, 200, 500 or 1000 nucleotides. The full-length sequence corresponding to the entire gene transcript may be used. The lengths can be 100-2000 nucleotides. The degree of identity of the sense and antisense sequences to the targeted transcript should be at least 85%, for example, at least 90% such as, 95-100%.

[0398] Exemplary small interfering RNA (“siRNA”) molecules comprise a nucleotide sequence that is identical to about 19-21 contiguous nucleotides of the target mRNA. For example, the siRNA sequence commences with the dinucleotide AA, comprises a GC-content of about 30-70% (for example, 30-60%, such as 40-60% for example about 45%-55%), and does not have a high percentage identity to any nucleotide sequence other than the target in the genome of the mammal in which it is to be introduced, for example as determined by standard BLAST search.

[0399] Aptamers

[0400] In another example, a compound is a nucleic acid aptamer (adaptable oligomer). Aptamers are single stranded oligonucleotides or oligonucleotide analogs that are capable of forming a secondary and / or tertiary structure that provides the ability to bind to a particular target molecule, such as a protein or a small molecule, e.g., G-CSF or G- CSFR. Thus, aptamers are the oligonucleotide analogy to antibodies. In general, aptamers comprise about 15 to about 100 nucleotides, such as about 15 to about 40 nucleotides, for example about 20 to about 40 nucleotides, since oligonucleotides of a length that falls within these ranges can be prepared by conventional techniques.

[0401] An aptamer can be isolated from or identified from a library of aptamers. An aptamer library is produced, for example, by cloning random oligonucleotides into a vector (or an expression vector in the case of an RNA aptamer), wherein the random sequence is flanked by known sequences that provide the site of binding for PCR primers. An aptamer that provides the desired biological activity (e.g., binds specifically to G- CSF or G-CSFR) is selected. An aptamer with increased activity is selected, for example, using SELEX (Sytematic Evolution of Ligands by Exponential enrichment). Suitable methods for producing and / or screening an aptamer library are described, for example, in Elloington and Szostak, Nature 346:818-22, 1990; US 5270163; and / or US 5475096.

[0402] Assaying Activity of a Compound

[0403] Binding to G-CSFR and Mutants Thereof It will be apparent to the skilled artisan from the disclosure herein that some compounds of the present disclosure bind to the ligand binding domain of hG-CSFR and to specific mutant forms of the ligand binding domain of hG-CSFR (e.g., SEQ ID NO: 1 without or with certain point mutations) and / or bind to both human and cynomolgus monkey G-CSFR. Methods for assessing binding to a protein are known in the art, e.g., as described in Scopes (In: Protein purification: principles and practice, Third Edition, Springer Verlag, 1994). Such a method generally involves labeling the protein and contacting it with immobilized compound. Following washing to remove non-specific bound protein, the amount of label and, as a consequence, bound protein is detected. Of course, the protein can be immobilized and the compound that inhibits G-CSF signaling labeled. Panning-type assays can also be used. Alternatively, or additionally, surface plasmon resonance assays can be used, such as a surface plasmon resonance assay in which the antibody or G-CSFR is immobilized on a solid substrate (i.e., a surface plasmon resonance chip).

[0404] Optionally, the dissociation constant (Kd) of a protein for hG-CSFR or an epitope thereof is determined. The "Kd" or "Kd value" for a hG-CSFR binding protein is determined. In some examples measured by a radiolabeled or fluorescently-labeled hG- CSFR binding assay. This assay equilibrates the protein with a minimal concentration of labeled G-CSFR in the presence of a titration series of unlabeled hG-CSFR. Following washing to remove unbound hG-CSFR, the amount of label is determined, which is indicative of the Kd of the protein.

[0405] According to another example the Kd or Kd value is measured by using surface plasmon resonance assays, e.g., using BIAcore surface plasmon resonance (BIAcore, Inc., Piscataway, NJ) with immobilized hG-CSFR or a region thereof.

[0406] In some examples, proteins having a similar Kd or a higher Kd than C 1.2 or C 1.2G are selected, because they are likely to compete for binding to hG-CSFR.

[0407] The assays described above can also be used to detect the level of binding of a compound to hG-CSFR or a ligand binding domain thereof (e.g., SEQ ID NO: 1) or mutant form thereof.

[0408] In one example, a protein of the present disclosure binds to a polypeptide of SEQ ID NO: 1 in which an alanine is substituted for the lysine at position 167 of SEQ ID NO: 1 and / or in which an alanine is substituted for the histidine at position 168 of SEQ ID NO: 1 at substantially the same level (e.g., within 10% or 5% or 1%) as it binds to SEQ ID NO: 1.

[0409] In one example, a protein of the present disclosure binds to a polypeptide of SEQ ID NO: 1 in which an alanine is substituted for the arginine at position 287 of SEQ ID NO: 1 at a level at least about 100 fold or 150 fold or 160 fold or 200 fold lower than it binds to a polypeptide of SEQ ID NO: 1. In one example, a protein of the present disclosure binds to a polypeptide of SEQ ID NO: 1 in which an alanine is substituted for the arginine at position 287 of SEQ ID NO: 1 at a level at least about 160 fold lower than it binds to a polypeptide of SEQ ID NO: 1.

[0410] In one example, a protein of the present disclosure binds to a polypeptide of SEQ ID NO: 1 in which an alanine is substituted for the histidine at position 237 of SEQ ID NO: 1 at a level at least about 20 fold or 40 fold or 50 fold or 60 fold lower than it binds to a polypeptide of SEQ ID NO: 1. In one example, a protein of the present disclosure binds to a polypeptide of SEQ ID NO: 1 in which an alanine is substituted for the histidine at position 237 of SEQ ID NO: 1 at a level at least about 50 fold lower than it binds to a polypeptide of SEQ ID NO: 1.

[0411] In one example, a protein of the present disclosure binds to a polypeptide of SEQ ID NO: 1 in which an alanine is substituted for the methionine at position 198 of SEQ ID NO: 1 at a level at least about 20 fold or 40 fold or 60 fold or 70 fold lower than it binds to a polypeptide of SEQ ID NO: 1. In one example, a protein of the present disclosure binds to a polypeptide of SEQ ID NO: 1 in which an alanine is substituted for the methionine at position 198 of SEQ ID NO: 1 at a level at least about 40 fold lower than it binds to a polypeptide of SEQ ID NO: 1.

[0412] In one example, a protein of the present disclosure binds to a polypeptide of SEQ ID NO: 1 in which an alanine is substituted for the tyrosine at position 172 of SEQ ID NO: 1 at a level at least about 20 fold or 30 fold or 40 fold lower than it binds to a polypeptide of SEQ ID NO: 1. In one example, a protein of the present disclosure binds to a polypeptide of SEQ ID NO: 1 in which an alanine is substituted for the tyrosine at position 172 of SEQ ID NO: 1 at a level at least about 40 fold lower than it binds to a polypeptide of SEQ ID NO: 1.

[0413] In one example, a protein of the present disclosure binds to a polypeptide of SEQ ID NO: 1 in which an alanine is substituted for the leucine at position 171 of SEQ ID NO: 1 at a level at least about 100 fold or 120 fold or 130 fold or 140 fold lower than it binds to a polypeptide of SEQ ID NO: 1. In one example, a protein of the present disclosure binds to a polypeptide of SEQ ID NO: 1 in which an alanine is substituted for the leucine at position 171 of SEQ ID NO: 1 at a level at least about 140 fold lower than it binds to a polypeptide of SEQ ID NO: 1.

[0414] In one example, a protein of the present disclosure binds to a polypeptide of SEQ ID NO: 1 in which an alanine is substituted for the leucine at a position 111 of SEQ ID NO: 1 at a level at least about 20 fold or 40 fold or 60 fold or 70 fold lower than it binds to a polypeptide of SEQ ID NO: 1. In one example, a protein of the present disclosure binds to a polypeptide of SEQ ID NO: 1 in which an alanine is substituted for the leucine at a position 111 of SEQ ID NO: 1 at a level at least about 60 fold lower than it binds to a polypeptide of SEQ ID NO: 1.

[0415] In one example, a protein of the present disclosure binds to a polypeptide of SEQ ID NO: 1 in which an alanine is substituted for the histidine at position 168 of SEQ ID NO: 1 at a level no more than 5 fold or 4 fold or 3 fold or 2 fold or 1 fold lower than it binds to a polypeptide of SEQ ID NO: 1.

[0416] In one example, a protein of the present disclosure binds to a polypeptide of SEQ ID NO: 1 in which an alanine is substituted for the lysine at position 167 of SEQ ID NO: 1 at a level no more than 5 fold or 4 fold or 3 fold or 2 fold or 1 fold lower than it binds to a polypeptide of SEQ ID NO: 1.

[0417] The level of binding is conveniently determined using a biosensor.

[0418] The present disclosure contemplates any combination of the foregoing characteristics. In one example, a protein described herein has all of the binding characteristics set forth in the preceding seven paragraphs.

[0419] Epitope Mapping

[0420] In another example, the epitope bound by a protein described herein is mapped. Epitope mapping methods will be apparent to the skilled artisan. For example, a series of overlapping peptides spanning the hG-CSFR sequence or a region thereof comprising an epitope of interest, e.g., peptides comprising 10-15 amino acids are produced. The protein is then contacted to each peptide and the peptide(s) to which it binds determined. This permits determination of peptide(s) comprising the epitope to which the protein binds. If multiple non-contiguous peptides are bound by the protein, the protein may bind a conformational epitope.

[0421] Alternatively, or in addition, amino acid residues within hG-CSFR are mutated, e.g., by alanine scanning mutagenesis, and mutations that reduce or prevent protein binding are determined. Any mutation that reduces or prevents binding of the protein is likely to be within the epitope bound by the protein.

[0422] A further method is exemplified herein, and involves binding hG-CSFR or a region thereof to an immobilized protein of the present disclosure and digesting the resulting complex with proteases. Peptide that remains bound to the immobilized protein are then isolated and analyzed, e.g., using mass spectrometry, to determine their sequence. A further method involves converting hydrogens in hG-CSFR or a region thereof to deutrons and binding the resulting protein to an immobilized protein of the present disclosure. The deutrons are then converted back to hydrogen, the hG-CSFR or region thereof isolated, digested with enzymes and analyzed, e.g., using mass spectrometry to identify those regions comprising deutrons, which would have been protected from conversion to hydrogen by the binding of a protein described herein.

[0423] Optionally, the dissociation constant (Kd) of a protein for hG-CSFR or an epitope thereof is determined. The "Kd" or "Kd value" for a hG-CSFR binding protein is in one example measured by a radiolabeled or fluorescently-labeled hG-CSFR binding assay. This assay equilibrates the protein with a minimal concentration of labeled G-CSFR in the presence of a titration series of unlabeled hG-CSFR. Following washing to remove unbound hG-CSFR, the amount of label is determined, which is indicative of the Kd of the protein.

[0424] According to another example the Kd or Kd value is measured by using surface plasmon resonance assays, e.g., using BIAcore surface plasmon resonance (BIAcore, Inc., Piscataway, NJ) with immobilized hG-CSFR or a region thereof.

[0425] In some examples, proteins having a similar Kd or a higher Kd than C 1.2 or C 1.2G are selected, because they are likely to compete for binding to hG-CSFR.

[0426] Determining Competitive Binding

[0427] Assays for determining a protein that competitively inhibits binding of monoclonal antibody Cl.2 or C1.2G will be apparent to the skilled artisan. For example, Cl.2 or C1.2G is conjugated to a detectable label, e.g., a fluorescent label or a radioactive label. The labeled antibody and the test protein are then mixed and contacted with hG- CSFR or a region thereof (e.g., a polypeptide comprising SEQ ID NO: 1) or a cell expressing same. The level of labeled Cl.2 or C1.2G is then determined and compared to the level determined when the labeled antibody is contacted with the hG-CSFR, region or cells in the absence of the protein. If the level of labeled Cl.2 or C1.2G is reduced in the presence of the test protein compared to the absence of the protein, the protein is considered to competitively inhibit binding of Cl.2 or C1.2G to hG-CSFR.

[0428] Optionally, the test protein is conjugated to different label to Cl.2 or C1.2G. This alternate labeling permits detection of the level of binding of the test protein to hG-CSFR or the region thereof or the cell.

[0429] In another example, the protein is permitted to bind to hG-CSFR or a region thereof (e.g., a polypeptide comprising SEQ ID NO: 1) or a cell expressing same prior to contacting the hG-CSFR, region or cell with Cl.2 or C1.2G. A reduction in the amount of bound C 1.2 or C1.2G in the presence of the protein compared to in the absence of the protein indicates that the protein competitively inhibits Cl.2 or C1.2G binding to hG- CSFR. A reciprocal assay can also be performed using labeled protein and first allowing Cl.2 or C1.2G to bind to G-CSFR. In this case, a reduced amount of labeled protein bound to hG-CSFR in the presence of Cl.2 or C1.2G compared to in the absence of Cl.2 or C1.2G indicates that the protein competitively inhibits binding of C1.2 or C1.2G to hG-CSFR.

[0430] Any of the foregoing assays can be performed with a mutant form of hG-CSFR and / or SEQ ID NO: 1 and / or a ligand binding region of hG-CSFR to which Cl.2 or C1.2G binds, e.g., as described herein.

[0431] Determining Neutralization

[0432] In some examples of the present disclosure, a compound is capable of neutralizing hG-CSFR signaling.

[0433] Various assays are known in the art for assessing the ability of a compound to neutralize signaling of a ligand through a receptor.

[0434] In one example, the compound that inhibits G-CSF signaling reduces or prevents G-CSF binding to the hG-CSFR. These assays can be performed as a competitive binding assay as described herein using labeled G-CSF and / or labeled protein.

[0435] In another example, the compound that inhibits G-CSF signaling reduces formation of CFU-G when CD34+bone marrow cells are cultured in the presence of G- CSF. In such assays, CD34+bone marrow cells are cultured in a semi-solid cell culture medium in the presence of G-CSF (e.g., about lOng / ml cell culture medium) and, optionally stem cell factor (e.g., about lOng / ml cell culture medium) in the presence or absence of a test compound. After a sufficient time for granulocyte clones (CFU-G) to form, the number of clones or colonies is determined. A reduction in the number of colonies in the presence of the compound that inhibits G-CSF signaling compared to in the absence of the compound that inhibits G-CSF signaling indicates that the compound that inhibits G-CSF signaling neutralizes G-CSF signaling. By testing multiple concentrations of the compound that inhibits G-CSF signaling an ICso is determined, i.e., a concentration at which 50% of the maximum inhibition of CFU-G formation occurs. In one example, the ICso is 0.2nM or less, such as O.lnM or less, for example, 0.09nM or less, or 0.08nM or less, or 0.07nM or less, or 0.06nM or less or0.05nM or less. In one example, the ICso is 0.04nM or less. In another example, the ICso is 0.02nM or less. The foregoing ICsos relate to any CFU-G assay described herein. In a further example, the compound that inhibits G-CSF signaling reduces proliferation of cells (e.g., BaF3 cells) expressing hG-CSFR which are cultured in the presence of G-CSF. Cells are cultured in the presence of G-CSF (e.g., 0.5ng / ml) and the presence or absence of a test compound. Methods for assessing cell proliferation are known in the art and include, for example, MTT reduction and thymidine incorporation. A compound that reduces the level of proliferation compared to the level observed in the absence of the compound is considered to neutralize G-CSF signaling. By testing multiple concentrations of the compound an ICso is determined, i.e., a concentration at which 50% of the maximum inhibition of cell proliferation occurs. In one example, the ICso is 6nM or less, such as 5.9nM or less. In another example, the IC50 is 2nM or less or InM or less or 0.7nM or cell or 0.6nM or less or 0.5nM or less. The foregoing ICsos relate to any cell proliferation assay described herein.

[0436] In a further example, the compound that inhibits G-CSF signaling reduces mobilization of hematopoietic stem cells and / or endothelial progenitor cells in vivo following G-CSF administration and / or reduces the number of neutrophils in vivo, e.g., following G-CSF administration (however this is not essential). For example, the compound that inhibits G-CSF signaling is administered, optionally before, at the time of or after administration of G-CSF or a modified form thereof (e.g., PEGylated G-CSF or filgrastim). The number of hematopoietic stem cells (e.g., expressing CD34 and / or Thyl) and / or endothelial progenitor cells (e.g., expressing CD34 and VEGFR2) and / or neutrophils (identified morphologically and / or expressing e.g., CD10, CD14, CD31 and / or CD88) is assessed. A compound that reduces the level of the cell(s) compared to the level observed in the absence of the compound is considered to neutralize G-CSF signaling. In one example, the compound that inhibits G-CSF signaling reduces the number of neutrophils without inducing neutropenia.

[0437] Other methods for assessing neutralization of G-CSF signaling are contemplated by the present disclosure.

[0438] Determining Effector Function

[0439] As discussed herein, some proteins of the present disclosure have reduced effector function. Methods for assessing ADCC activity are known in the art.

[0440] In one example, the level of ADCC activity is assessed using a51Cr release assay, an europium release assay or a35S release assay. In each of these assays, cells expressing G-CSFR are cultured with one or more of the recited compounds that inhibit G-CSF signaling for a time and under conditions sufficient for the compound to be taken up by the cell. In the case of a35S release assay, cells expressing hG-CSFR can be cultured with35S-labeled methionine and / or cysteine for a time sufficient for the labeled amino acids to be incorporated into newly synthesized proteins. Cells are then cultured in the presence or absence of the protein and in the presence of immune effector cells, e.g., peripheral blood mononuclear cells (PBMC) and / or NK cells. The amount of51Cr, europium and / or35S in cell culture medium is then detected, and little or no change in the presence of the protein compared to in the absence of protein (or a reduced level of the compound compared to the level observed in the presence of an anti-hG-CSFR antibody comprising a human IgGl Fc) indicates that the protein has reduced effector function. Exemplary publications disclosing assays for assessing the level of ADCC induced by a protein include Hellstrom, et al. Proc. Natl Acad. Sci. USA 53:7059-7063, 1986 and Bruggemann, et al., J. Exp. Med. 166: 1351- 1361 , 1987.

[0441] Other assays for assessing the level of ADCC induced by a protein include ACTI™ nonradioactive cytotoxicity assay for flow cytometry (CellTechnology, Inc. CA, USA) or CytoTox 96® non-radioactive cytotoxicity assay (Promega, WI, USA).

[0442] Clq binding assays may also be carried out to confirm that the protein is able to bind Clq and may induce CDC. To assess complement activation, a CDC assay may be performed (see, for example, Gazzano-Santoro et al, J. Immunol. Methods 202: 163, 1996.

[0443] Determining Half Life

[0444] Some proteins encompassed by the present disclosure have an improved half-life, e.g., are modified to extend their half-life compared to proteins that are unmodified.

[0445] Methods for determining a protein with an improved half-life will be apparent to the skilled person. For example, the ability of a protein to bind to a neonatal Fc receptor

[0446] (FcRn) is assessed. In this regard, increased binding affinity for FcRn increased the serum half-life of the molecule (see for example, Kim et al., Eur J Immunol., 24:2429,

[0447] 1994).

[0448] The half-life of a protein of the disclosure can also be measured by pharmacokinetic studies, e.g., according to the method described by Kim et al, Eur J of

[0449] Immunol 24:542, 1994. According to this method radiolabeled protein is injected intravenously into mice and its plasma concentration is periodically measured as a function of time, for example at 3 minutes to 72 hours after the injection. The clearance curve thus obtained should be biphasic, that is, an alpha phase and beta phase. For the determination of the in vivo half-life of the protein, the clearance rate in beta-phase is calculated and compared with that of the wild type or unmodified protein. Efficacy

[0450] The efficacy of a compound that inhibits G-CSF signaling to reduce, prevent, inhibit or delay development of a neutrophil-associated condition can be assessed by comparing the degree of severity of the disease or symptoms in subjects administered with the compound relative to subjects not administered the compound. Alternatively, or additionally, therapeutic efficacy of candidate compounds can be assessed in an animal model specific for the condition being assessed.

[0451] Methods for assessing the efficacy of treatment of a compound of the disclosure for a neutrophil- associated condition will be apparent to the skilled person and / or described herein.

[0452] For example, the efficacy of treatment of HS can be measured by assessing the number of abscesses, nodules, and draining tunnels (fistulae or sinus tracts). These numbers can be used to derive the IHS4 and other response criteria (HiSCR50, HiSCR75, Modified HiSCR, and IHS4-55). Efficacy of treatment can also be assessed according to patient-reported outcomes such as a Dermatology Life Quality Index (DLQI), pain numerical rating scale (NRS) score, and / or Hidradenitis Suppurativa Quality of Life (HiSQOL).

[0453] In one example, the efficacy of treatment of PPP can be measured by assessing the ppPASI score and / or the palm-sole PGA.

[0454] In some examples, assessing the efficacy of a compound of the disclosure comprises detecting and / or quantifying the level of expression of a biomarker, such as an inflammatory cytokine as described herein, in the subject. Detecting and / or quantifying biomarkers can be performed by any method known in the art. For instance, in some examples, the levels of biomarkers are assessed using mass spectrometry. The mass spectrometry may be performed in conjunction with ultra- performance liquid chromatography (UPLC), high-performance liquid chromatography (HPLC), gas chromatography (GC), gas chromatography / mass spectroscopy (GC / MS), and UPLC, for example. Other methods of assessing levels of biomarkers include biological methods, such as but not limited to ELISA assays, Western Blot and multiplexed immunoassays etc. Other techniques may include using quantitative arrays, PCR, Northern Blot analysis. To determine levels of components or factors, it is not necessary that an entire component, e.g., a full-length protein or an entire RNA transcript, be present or fully sequenced. In other words, determining levels of, for example, a fragment of protein being analyzed may be sufficient to conclude or assess that the level of the biomarker being analyzed is increased or decreased. Similarly, if, for example, arrays or blots are used to determine component levels, the presence / absence / strength of a detectable signal may be sufficient to assess levels of biomarkers.

[0455] To assess levels of biomarkers, a sample may be taken from the subject. The sample may or may not processed prior assaying levels of the components of the biomarker profile. For example, whole blood may be taken from an individual and the blood sample may be processed, e.g., centrifuged, to isolate plasma or serum from the blood. The sample may or may not be stored, e.g., frozen, prior to processing or analysis.

[0456] Biological samples that may be tested in a method of the invention include whole blood, blood serum, plasma, urine, saliva, or other bodily fluid (stool, tear fluid, synovial fluid, sputum), or an extract or purification therefrom, or dilution thereof. Biological samples also include tissue homogenates, tissue sections and biopsy specimens from a live subject, or taken post-mortem. The samples can be prepared, for example where appropriate diluted or concentrated, and stored in the usual manner.

[0457] Compositions

[0458] In some examples, a compound as described herein can be administered orally, parenterally, by inhalation spray, adsorption, absorption, topically, rectally, nasally, bucally, vaginally, intraventricularly, via an implanted reservoir in dosage formulations containing conventional non-toxic pharmaceutically-acceptable carriers, or by any other convenient dosage form. The term “parenteral” as used herein includes subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal, intraventricular, intrastemal, and intracranial injection or infusion techniques. In one example, the compound as described herein is administered subcutaneously.

[0459] Methods for preparing a compound into a suitable form for administration (e.g. a pharmaceutical composition) are known in the art and include, for example, methods as described in Remington's Pharmaceutical Sciences (18th ed., Mack Publishing Co., Easton, Pa., 1990) and U.S. Pharmacopeia: National Formulary (Mack Publishing Company, Easton, Pa., 1984).

[0460] The pharmaceutical compositions of this disclosure are particularly useful for parenteral administration, such as intravenous administration or subcutaneous administration or administration into a body cavity or lumen of an organ or joint. The compositions for administration will commonly comprise a solution of the compound that inhibits G-CSF signaling dissolved in a pharmaceutically acceptable carrier, for example an aqueous carrier. A variety of aqueous carriers can be used, e.g., buffered saline and the like. The compositions may contain pharmaceutically acceptable auxiliary substances as required to approximate physiological conditions such as pH adjusting and buffering agents, toxicity adjusting agents and the like, for example, sodium acetate, sodium chloride, potassium chloride, calcium chloride, sodium lactate and the like. The concentration of compound of the present disclosure in these formulations can vary widely, and will be selected primarily based on fluid volumes, viscosities, body weight and the like in accordance with the particular mode of administration selected and the patient's needs. Exemplary carriers include water, saline, Ringer's solution, dextrose solution, and 5% human serum albumin. Nonaqueous vehicles such as mixed oils and ethyl oleate may also be used. Liposomes may also be used as carriers. The vehicles may contain minor amounts of additives that enhance isotonicity and chemical stability, e.g., buffers and preservatives.

[0461] Upon formulation, compounds of the present disclosure will be administered in a manner compatible with the dosage formulation and in such amount as is therapeutically / prophylactically effective. Formulations are easily administered in a variety of dosage forms, such as the type of injectable solutions described above, but other pharmaceutically acceptable forms are also contemplated, e.g., tablets, pills, capsules or other solids for oral administration, suppositories, pessaries, nasal solutions or sprays, aerosols, inhalants, liposomal forms and the like. Pharmaceutical "slow release" capsules or compositions may also be used. Slow release formulations are generally designed to give a constant drug level over an extended period and may be used to deliver compounds of the present disclosure.

[0462] One skilled in the art would be able, by routine experimentation, to determine what an effective dose of the compound of the disclosure would be for the purpose of treating or preventing or delaying progression or reducing or inhibiting or hindering development of a neutrophil-associated condition e as described herein. For example, a therapeutically active amount of a compound may vary according to factors such as the disease stage, age, sex and weight of the subject, and the ability of the compound to elicit a desired response in the subject. The dosage regimen may be adjusted to provide the optimum therapeutic response. For example, the optimum therapeutic response may be a reduction in frequency of a symptom of a neutrophil-associated condition.

[0463] In one example, several divided doses may be administered daily, or the dose may be proportionally reduced as indicated by the exigencies of the therapeutic situation. Generally, however, an effective dosage is expected to be in the range of about 1 to 200 mg / kg body weight. Furthermore, an effective dosage is expected to be administered at least one or more times, such as every 7-30 days, such as every 10-22 days, for example, every 10-15 days. Administration of a compound according to the methods of the present disclosure can be continuous or intermittent, depending, for example, on the subject’s physiological condition, whether the purpose of the administration is therapeutic or prophylactic, and other factors known to skilled practitioners. For example, the compound described herein may be administered prior to, during or after development of a complication as described herein, or pro re nata, In one example, the compound is administered pro re nata. For example, the compound is administered at the onset of a neutrophil-associated condition, or a symptom thereof.

[0464] Combination Therapies

[0465] In some examples, the compound is administered in combination with another therapy for treating a disease or condition described herein, either as combined or additional treatment steps or as additional components of a therapeutic formulation.

[0466] For example, the other therapy may comprise administration of another compound, such as an anti-inflammatory compound, e.g., methotrexate or a non-steroidal anti-inflammatory compound. Alternatively, or additionally, the other compound may be an immunosuppressant. Alternatively, or additionally, the other compound may be a corticosteroid, such as prednisone and / or prednisolone. In on example, the other compound is methotrexate. Alternatively, or additionally, the other compound may be cyclophosphamide.

[0467] In some examples, the compound is administered simultaneously with the other compound. In some examples, the compound that inhibits G-CSF signaling is administered before the other compound. In one example, the compound that inhibits G- CSF signaling is administered after the other compound.

[0468] In some examples, the compound is administered in combination with a cell. In some examples, the cell is a stem cell, such as a mesenchymal stem cell. In some examples, the compound is administered in combination with a gene therapy.

[0469] In some examples, the compound is administered in combination with a nonpharmaceutical intervention, for example, apharesis, such as plasmapheresis, cytapheresis, leukapheresis, granulocyte and / or monocyte apheresis. In this context, the antibody can be administered during the period of time in which the non-pharmaceutical intervention is being performed and will be considered “in combination with” the non- pharmaceutical intervention. For example, the non-pharmaceutical intervention may be granulocyte and / or monocyte apheresis, which is performed once per week for five weeks and the compound can be administered over this time period. In one example, the compound is administered before the non-pharmaceutical intervention. In one example, the compound is administered after the non-pharmaceutical intervention.

[0470] Another non-pharmaceutical intervention is light therapy. As is known in the art, light therapy is used to treat some neutrophilic dermatoses.

[0471] Kits

[0472] Another example of the disclosure provides kits containing compounds useful for treating or delaying progression or reducing or inhibiting or hindering development of a neutrophil-associated condition in a subject as described above.

[0473] In one example, the kit comprises (a) a container comprising a compound that inhibits G-CSF signaling as described herein, optionally in a pharmaceutically acceptable carrier or diluent; and (b) a package insert with instructions for treating or delaying progression or reducing or inhibiting or hindering development of a neutrophil- associated condition in a subject, wherein the subject has had an inadequate clinical response to an anti-TNF therapy for the neutrophil-associated condition.

[0474] In accordance with this example of the disclosure, the package insert is on or associated with the container. Suitable containers include, for example, bottles, vials, syringes, etc. The containers may be formed from a variety of materials such as glass or plastic. The container holds or contains a composition that is effective for treating or delaying progression or reducing or inhibiting or hindering development of a neutrophil- associated condition in a subject, wherein the subject has had an inadequate clinical response to an anti-TNF therapy for the neutrophil-associated condition and may have a sterile access port (for example, the container may be an intravenous solution bag or a vial having a stopper pierceable by a hypodermic injection needle). At least one active agent in the composition is the compound that inhibits G-CSF signaling. The label or package insert indicates that the composition is used for treating or delaying progression or reducing or inhibiting or hindering development of a neutrophil- associated condition in a subject eligible for treatment (i.e., the subject has had an inadequate clinical response to an anti-TNF therapy for the neutrophil-associated condition), with specific guidance regarding dosing amounts and intervals of compound and any other medicament being provided. The kit may further comprise an additional container comprising a pharmaceutically acceptable diluent buffer, such as bacteriostatic water for injection (BWFI), phosphate-buffered saline, Ringer's solution, and / or dextrose solution. The kit may further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles, and syringes. SEQUENCES OF THE DISCLOSURE

[0475] The present disclosure includes the following non-limiting Examples.

[0476] EXAMPLES

[0477] Example 1: A Multicenter, Open-label, 2-regimen, Repeat-dose Study to Assess the Safety and Pharmacokinetics of Intravenous CSL324 in Subjects with Hidradenitis Suppurativa and Palmoplantar Pustulosis (CSL324_1002)

[0478] Objectives

[0479] Primary Objective: To evaluate the safety of repeat doses of intravenous (IV) CSL324 in subjects with hidradenitis suppurativa (HS) and palmoplantar pustulosis (PPP).

[0480] Secondary Objectives:

[0481] 1. To determine the pharmacokinetics (PK) of repeat doses of IV CSL324

[0482] 2. To investigate the immunogenicity of repeat doses of IV CSL324

[0483] Exploratory Objectives

[0484] 1. To measure the effect of CSL324 on the inflammatory cell infiltrate, including neutrophilic infiltration, in skin lesion biopsies

[0485] 2. To explore the pharmacodynamic (PD) properties and potential biomarkers of CSL324

[0486] 3. To assess the potential efficacy, including patient-reported outcomes, of repeat doses of IV CSL324

[0487] 4. To explore potential PK / PD / efficacy relationships

[0488] Methodology

[0489] This was a phase lb, open-label, multicenter study for the treatment of subjects with HS and PPP > 18 and < 75 years of age at the time of providing written informed consent. The study consisted of a 28-day Screening Period, a 15-week Treatment Period, and a 9-week Follow-up Period. Subjects with HS or PPP were screened from Day - 28 to Day - 2.

[0490] Subjects were administered CSL324 on Days 1, 22, 43, 64, and 85 following hematology assessments the day before administration to measure their absolute neutrophil count (ANC). On the day of CSL324 administration, subjects were required to remain at the study site for at least 1 hour after the end of infusion for safety observations. Subjects were required to attend the study site weekly and more frequently during the first week after each CSL324 administration for safety, PK, PD, and / or efficacy, including patient-reported assessments. Skin lesion biopsies were collected on Day 1 (before CSL324 was administered) and at the end of the Treatment Period (on Day 105 or, for subjects who discontinued treatment, at 3 weeks after their last dose of CSL324 was administered). All subjects were required to self-monitor their oral temperature daily. During the 9-week Follow-up Period, subjects were required to attend 2 Follow-up Visits and the End of Study (EOS) Visit on Day 168 (± 5 days) for safety, PK, immunogenicity, PD, and efficacy assessments.

[0491] The study consisted of 2 cohorts (Cohort 1 [0.3 mg / kg] and Cohort 2 [0.6 mg / kg]). Each cohort planned to enrol 10 subjects with HS and 10 subjects with PPP (ie, 20 subjects total in each cohort). Enrolment of subjects was sequential by indication across Cohorts 1 and 2. Enrolment of subjects began in Cohort 1. The overall safety of the study was overseen by a Safety Review Committee, including the recommendation regarding the dose to be administered to subjects enrolled in Cohort 2 and investigation of and recommendation following events triggering stopping criteria. When at least 10 subjects with a given indication (either HS or PPP) were enrolled in Cohort 1 and the exact dose of CSL324 to be administered to subjects in Cohort 2 had been determined, enrolment of subjects with that indication began in Cohort 2. Enrolment of subjects with the other indication continued in Cohort 1 until 10 subjects had been enrolled, resulting in 20 subjects planned in total for Cohort 1. Ten subjects with a given indication must have been enrolled in Cohort 1 before any subject with that indication could have been enrolled in Cohort 2.

[0492] Number of Subjects

[0493] Planned: 40 subjects total (20 subjects per indication).

[0494] Actual: 39 subjects total (22 subjects with HS and 17 subjects with PPP).

[0495] Diagnosis and Main Criteria for Inclusion: To be eligible for this study, male or female subjects must have been capable of providing written informed consent and must have been between 18 and 75 years of age, inclusive, at the time of providing written informed consent. Subjects with HS and subjects with PPP must have had confirmed clinical diagnosis of HS and PPP, respectively, at least 6 months before Screening. Subjects with HS must have had moderate to severe HS as per ISH4 guidelines at Screening and inadequate response to at least a 3-month (90 days) trial of oral antibiotics for treatment of HS (or demonstrated intolerance to, or had a contraindication to, oral antibiotics for treatment of their HS). Subjects with PPP must have had PPP differentiated from other forms of pustulosis or psoriasis with a Palmoplantar Pustulosis Psoriasis Area and Severity Index (ppPASI) score of > 12 at Screening and inadequate response to previous therapy for the treatment of PPP. All subjects must, in the opinion of the investigator, understand the nature, scope, and possible consequences of the study.

[0496] Investigational Product, Dose and Mode of Administration, Batch Number(s)

[0497] In this study, 2 dose levels of CSL324 were administered IV to subjects with HS or PPP to further explore the potential PK / PD (ANC) relationship of CSL324. The dosing regimen for Cohort 1, 5 doses of 0.3 mg / kg CSL324 administered at 21 -day intervals, was selected based on safety, PK, and PD data from the phase 1 Study CSL324_1001 and data from the nonclinical Good Laboratory Practice toxicity study in cynomolgus monkeys (Study APQ0045).

[0498] CSL324 is a sterile solution for injection containing 10 mg / mL in 10 mL vials. Batch numbers used during the study were P100083750, P100196447, P100243340, P100270672, P100386596, and P100392537.

[0499] Duration of Treatment:

[0500] The duration of the treatment for each subject was 15 weeks, followed by a 9- week Follow-up Period.

[0501] Criteria for Evaluation

[0502] Primary Endpoint:

[0503] The primary endpoint for this study was treatment-emergent adverse events (TEAEs), including the adverse events of special interest (AESIs) Grades 3 and 4 Neutropenia and Grades 3 and 4 Infection.

[0504] Secondary Endpoints:

[0505] The secondary endpoints for this study were as follows: • Serum PK parameters of CSL324 for the first dose administered including the following: o Maximum concentration (C max) o Time to reach C max (T max) o Area under the concentration-time curve during a dosage interval (AUCtau)

[0506] • Serum PK parameters of CSL324 for the last dose administered including the following:

[0507] O Cmax

[0508] O Tmax

[0509] O AUCtau o Terminal elimination half-life o Total systemic clearance after IV dosing o Volume of distribution after IV dosing during the terminal elimination phase

[0510] • Trough concentration of CSL324 for each dose of CSL324 administered

[0511] • Accumulation ratio for AUCtau (the ratio between the AUCtau of the last dose and of the first dose) and for Cmax (the ratio between the Cmax of the last dose and of the first dose)

[0512] • Presence of anti-CSL324 antibodies in serum

[0513] Exploratory Endpoints:

[0514] The exploratory endpoints for this study were as follows:

[0515] • Cellular infiltrate in skin lesion biopsies assessed by: o Histology (immunohistochemistry and hematoxylin and eosin staining) o Transcriptome profiles from skin lesion biopsies were analyzed and reported separately

[0516] • Serial ANC profile in whole blood

[0517] • Serum concentration of: o Cytokines / chemokines (eg, granulocyte colony- stimulating factor [G- CSF], granulocyte-macrophage colony-stimulating factor) o Other selected biomarkers (eg, C-reactive protein, C3a, and C5a)

[0518] • Genetic profile from peripheral whole blood (optional) was analyzed and reported separately

[0519] • Transcriptome profile, including the neutrophil signature, from peripheral whole blood was analyzed and reported separately

[0520] • Neutrophil profile shift in peripheral blood films • Efficacy, including patient-reported outcome measures: o All subjects:

[0521] ■ Dermatology Life Quality Index (DLQI) score o Subjects with HS only:

[0522] ■ Hidradenitis Suppurativa Clinical Response (HiSCR)

[0523] ■ International Hidradenitis Suppurativa Severity Score System (IHS4)

[0524] ■ HS Physician Global Assessment (HS-PGA) score

[0525] ■ Pain numerical rating scale (NRS) score o Subjects with PPP only:

[0526] ■ ppPASI score

[0527] ■ PPP Physician Global Assessment (PPP-PGA) score

[0528] • PK / PD / efficacy relationships with appropriate biomarkers were explored and reported separately.

[0529] Statistical Methods

[0530] The sample size for this study was based upon empirical considerations; no formal sample size calculation was performed.

[0531] Analysis Populations:

[0532] The Enrolled Analysis Set comprised all subjects who provided written informed consent and who were eligible for inclusion in the study at Screening. Screening failures were not included in the Enrolled Analysis Set.

[0533] The Safety Analysis Set comprised all subjects in the Enrolled Analysis Set who received at least 1 partial dose of CSL324.

[0534] The Efficacy Analysis Set comprised all subjects in the Safety Analysis Set who had the at least 1 Baseline efficacy assessment and at least 1 matching / corresponding efficacy assessment after the first dose of CSL324. This was the primary Analysis Set for the analysis of each efficacy data.

[0535] The Per-Protocol Analysis Set comprised all subjects in the Efficacy Analysis Set who received at least 4 doses of CSL324 and at least 1 efficacy assessment after the fourth dose of CSL324 and had no major protocol deviations.

[0536] The PK Analysis Set comprised all subjects in the Safety Analysis Set who had at least 1 quantifiable concentration of CSL324 after administration of CSL324.

[0537] The PD Analysis Set comprised all subjects in the Safety Analysis Set who had PD data from before administration and from at least 1 time point after infusion of CSL324. Data Handling Conventions:

[0538] Missing data were indicated using a “blank” in subject listing displays. Answers such as “Not applicable” and “Not evaluable” were not considered to be missing data and were displayed as such. Subjects with the designation of treatment relationship for adverse events (AEs) and serious AEs (SAEs) missing had the worst case assumed to impute the relationship; if relationship to CSL324 was missing, it was assumed to be “Yes,” provided the event started on or after first CSL324 administration. There was no other imputation for missing data other than for partial dates and times.

[0539] Changes to subjects’ visits caused by the coronavirus disease 2019 (CO VID- 19) pandemic were captured for each subject in the electronic case report form (eCRF), on the “Visit Status” form. The eCRF page included different options for the primary visit modality, as well as whether the missed visit / alternate visit modality was due to COVID- 19. Changes to visit modality may have been a protocol deviation or could have been permitted via a contingency amendment; in either case, these data were captured. Since assessments (eg, vital signs, laboratory data) collected at each visit were known, the data missing due to COVID- 19 were determined. Assessments that were missed or required alternate visit modality (eg, televisits or home health visits) due to COVID- 19 were summarized. In addition, the number of subjects with missed visits or alternate visit modality, by visit, was summarized and data were listed.

[0540] Partially or completely missing dates were not imputed.

[0541] Pharmacokinetic and Pharmacodynamic Analyses:

[0542] Summary statistics were presented for concentration-time data for the PK Analysis Set. PK parameters of Cmax, AUCtau, area under the concentration-time curve to the last measurable plasma concentration (AUCiast), terminal elimination half-life (ti / 2), total systemic clearance (CLtot), volume of distribution after IV dosing during the terminal elimination phase (Vz), trough concentration (Ctrough), and Tmax for CSL324 were summarized with descriptive statistics by treatment group and study indication for the PK Analysis Set.

[0543] Biomarker parameters were summarized by treatment group and study indication for the PD Analysis Set by descriptive statistics.

[0544] Pearson and Spearman correlation analyses were applied to selected pairs of PK, PD, and efficacy parameters to explore the relationship between different components, especially for the changes associated with CSL324 administration.

[0545] Efficacy Analyses:

[0546] Efficacy parameters were summarized using descriptive statistics: • For subjects with HS: count of abscesses, nodules (total [sum of noninflammatory, and inflammatory], noninflammatory, and inflammatory), draining fistula count, HiSCR response rate (defined as a 50% reduction from Baseline in the total abscess and nodule count with no increase in abscess or draining fistula count), IHS4, and pain NRS

[0547] • For subjects with PPP: ppPASI score

[0548] • For both subjects with HS and PPP: disease-specific PGA and DLQI (total and subscores)

[0549] Safety Analyses:

[0550] All safety analyses were based on the Safety Analysis Set, and summaries included all scheduled safety assessments.

[0551] The extent of exposure to CSL324 was summarized by treatment group and study indication using descriptive statistics.

[0552] The observation period of AEs started from signing the written informed consent and ended with the EOS Visit. All AEs regardless of when they were reported were listed. TEAEs were defined as AEs with an onset at or after the start of first dose of study drug. Severity of AEs was assigned as mild, moderate, or severe. AEs were coded and grouped by system organ class (SOC) and Preferred Term using the Medical Dictionary for Regulatory Activities (MedDRA) Version 25.1. Severity of AEs was assigned according to the Common Terminology Criteria for Adverse Events (CTCAE), Version 5.0. In the summaries of TEAEs, subjects with multiple occurrences of the same TEAE were counted once.

[0553] AEs associated with COVID- 19, which can include a clinically significant laboratory finding like a positive test result for COVID- 19, were reported by investigators following reporting requirements outlined in the protocol. COVID- 19- associated AEs were identified via MedDRA coding. All COVID- 19-associated AEs were included in standard AE tables. Separate summaries of COVID- 19-associated AEs were also produced in the same way as for all AEs.

[0554] AESIs were defined as moderate (or CTCAE Grade 3) and severe (or CTCAE Grade 4) Neutropenia and moderate and severe (CTCAE Grade 3 and 4) Infection.

[0555] Hematology, biochemistry, and urinalysis tests; vital signs; and electrocardiogram summaries were summarized by visit and treatment group and study indication. Results of assessments of presence of anti-CSL324 antibodies in serum were summarized by a frequency table by visit, treatment group, and study indication.

[0556] Results Subject Disposition:

[0557] A total of 39 subjects were enrolled. In total, 27 subjects (69.2%) completed the treatment period, and 12 subjects (30.8%) discontinued CSL324 during the treatment period; 2 subjects (5.1%) discontinued CSL324 due to an AE.

[0558] Protocol Deviations:

[0559] Most protocol deviations were deemed not to have impacted the interpretation of the endpoints collected or the conclusions drawn and did not have a significant impact on subjects’ inclusion in the PP Analysis Set. No subjects were excluded from the Safety Analysis Set due to protocol deviations.

[0560] Demographics and Other Baseline Characteristics:

[0561] Overall, there was a greater proportion of female subjects versus male subjects (82.1% versus 17.9%). The majority of subjects were White (89.7%) and current smokers (69.2%). The 2 cohorts were not balanced; there were differences in baseline characteristics that reflect the small sample size and lack of randomization in this study. Within subjects with HS, those in Cohort 1 had a mean (range) IHS4 score of 38.4 (7 to 87), and 90.9% of subjects had severe HS; and those in Cohort 2 had a mean (range) IHS4 score of 18.6 (4 to 34), and 72.7% of subjects had severe HS. Within subjects with HS, subjects in Cohort 1 had a higher mean number of draining fistulas (5.7) than those in Cohort 2 (2.3). Within subjects with HS, 10 subjects (90.9%) in Cohort 1 and 5 subjects (45.5%) in Cohort 2 were current smokers; within subjects with PPP, 9 subjects (81.8%) in Cohort 1 and 3 subjects (50.0%) in Cohort 2 were current smokers. Within subjects with HS, 2 subjects (18.2%) in Cohort 1 and 5 subjects (45.5%) in Cohort 2 reported prior biologic use; within subjects with PPP, 1 subject (9.1%) in Cohort 1 and 1 subject (16.7%) in Cohort 2 reported prior biologic use.

[0562] Overall, 17 subjects (43.6%) recorded intake of at least 1 prior medication. Within subjects with HS, 4 subjects in Cohort 1 (36.4%) and 4 subjects in Cohort 2 (36.4%) took prior medications; within subjects with PPP, 7 subjects in Cohort 1 (63.6%) and 2 subjects in Cohort 2 (33.3%) took prior medications.

[0563] Overall, 39 subjects (100.0%) recorded intake of at least 1 concomitant medication.

[0564] Pharmacokinetics Results:

[0565] • For subjects with HS and PPP in Cohorts 1 and 2, mean serum concentration of CSL324 peaked at 1 hour after the start of infusion and then decreased to very low or undetectable levels by Day 21. Subsequent infusions exhibited similar PK behavior. • Following the first and last doses, mean Cmax increased in a dose-proportional manner; however, the increase in mean AUCiast was greater than doseproportional. The serum PK parameters of CSL324 were generally similar between subjects with PPP and HS in both cohorts.

[0566] • Overall, there was minimal to no accumulation of CSL324 in Cohort 1 and minimal accumulation of CSL324 in Cohort 2 based on AUCtau ratio of the last dose to the first dose of 1.67 (95% confidence interval: 1.255, 2.214) and 1.49 (95% confidence interval: 0.753, 2.933) in subjects with HS and subjects with PPP, respectively.

[0567] Pharmacodynamics Results:

[0568] • Following the first infusion on Day 1, mean ANC in Cohort 1 dropped to the nadir on Day 3 and then recovered to slightly below the Screening value on Day 14. Subsequent infusions followed a similar trend. Mean ANC in Cohort 2 followed a similar trend but with a greater decrease and slower recovery; mean ANC continued to stay suppressed and did not recover to the Screening value prior to the next infusion.

[0569] • G-CSF concentrations suggest that receptor occupancy was not maintained in Cohort 1 throughout the dosing period. In Cohort 2, G-CSF data suggest receptor occupancy was maintained in some but not all subjects during the dosing period.

[0570] • On Day 105, the percent change from Baseline of the mean C-reactive protein at Baseline in subjects with HS was 36.295% in Cohort 1 and 5.624% in Cohort 2. In subjects with PPP, the percent change was 18.171% in Cohort 1 and - 15.874% in Cohort 2.

[0571] • On Day 105, the percent change from Baseline of mean calprotectin in subjects with HS was 46.812% in Cohort 1 and -59.411% in Cohort 2. In subjects with PPP, the percent change from Baseline on Day 105 was 89.324% in Cohort 1 and 125.624% in Cohort 2.

[0572] • On Day 105, the percent change from Baseline of the mean pathologist score of relative neutrophil abundance (based on myeloperoxidase staining) in skin biopsy samples in subjects with HS was 73.469% in Cohort 1 and 41.865% in Cohort 2. In subjects with PPP, the percent change from Baseline on Day 105 was 50.000% in Cohort 1 and 111.111% in Cohort 2.

[0573] Efficacy Results:

[0574] • Efficacy assessments were exploratory endpoints in this study. The following trends in clinical responses were observed. • In subjects with HS, 11 subjects (57.9%) overall met the HiSCR criteria on Day 105 (7 [70.0%] subjects in Cohort 1 and 4 [44.4%] subjects in Cohort 2). The decrease in mean IHS4 score from Baseline to Day 105 was 11.8 (40.32%) (19.9 [51.67%] and 2.9 [27.71%] in Cohorts 1 and 2, respectively). Overall, 6 out of 22 (27.3%) subjects had an HS-PGA score of 0 to 3 and a > 2-point reduction from Baseline on Day 105 (5 out of 11 [45.5%] in Cohort 1 and 1 out of 11 [9.1%] in Cohort 2). No significant changes were observed in pain NRS score from Baseline to Day 105. The mean change (standard deviation) from Baseline on Day 105 in DLQI total score was -4.2 (6.35) overall (-4.9 [6.47] for Cohort 1 and -3.3 [6.50] for Cohort 2).

[0575] • In subjects with PPP, the decrease in mean ppPASI score from Baseline to Day 105 was 10.78 (40.39%) (16.28 [50.98%] and 2.53 [24.51%] in Cohorts 1 and 2, respectively). On Day 105, 1 out of 17 subjects (5.9%) (Cohort 1) had a PPP- PGA score of 0 or 1 and a > 2-point reduction from Baseline; no subjects in Cohort 2 had a score of 0 or 1 and a > 2-point reduction from Baseline. The mean change (standard deviation) from Baseline on Day 105 in DLQI total score was - 1.9 (6.32) overall (-1.7 [6.84] for Cohort 1 and -2.2 [6.08] for Cohort 2).

[0576] Safety Results:

[0577] • Overall, 33 subjects (84.6%) experienced 198 TEAEs, of which 70 TEAEs in 16 subjects (41.0%) were related to CSL324.

[0578] • Overall, the most common (> 10% of subjects) TEAEs were Headache, Fatigue, Neutropenia, PPP, Nausea, Hidradenitis, Pyrexia, and COVID- 19. In subjects with HS, the most common TEAEs were Headache, Fatigue, Hidradenitis, Pyrexia, and Nausea. In subjects with PPP, the most common TEAEs were PPP, Headache, Fatigue, Nausea, Back pain, Neutropenia, COVID- 19, Hidradenitis, Abdominal pain, Procedural pain, and Type 2 diabetes mellitus.

[0579] • Overall, the most common (> 10% of subjects) CSL324-related TEAEs were Fatigue, Neutropenia, Headache, and Nausea. In subjects with HS, the most common CSL324-related TEAEs were Fatigue and Nausea. In subjects with PPP, the most common CSL324-related TEAEs were Neutropenia, Fatigue, and Headache.

[0580] • There were no dose-dependent increases in the incidence of AEs except with Neutropenia, which was reported in 0 subjects in Cohort 1 and 7 subjects (41.2%) in Cohort 2.

[0581] • Of the 7 subjects who experienced a TEAE of Neutropenia, 5 subjects (12.8%) experienced Neutropenia with worst severity of mild, and 2 subjects (5.1%) experienced Neutropenia with worst severity of moderate. All TEAEs of Neutropenia were reported as related to CSL324, resolved spontaneously, and did not require treatment.

[0582] • The treatment-emergent AESIs reported were 4 events of moderate Neutropenia (2 subjects [5.1%]; these subjects were the same 2 subjects with a TEAE of moderate Neutropenia in the previous bullet) and 1 event of moderate Pyrexia (1 subject [2.6%]). While Pyrexia is not considered an AESI, it has been reported as such because it occurred in conjunction with an AESI of Grade 3 infection. The nature or site of the infection was not reported by the site. The 1 event of Pyrexia was considered to be not related to the administration of CSL324 by the investigator. All treatment-emergent AESIs occurred in Cohort 2. The proportion of subjects with a treatment-emergent AESI of Neutropenia was similar between subjects with HS and subjects with PPP (4.5% versus 5.9%, respectively).

[0583] • There were no deaths in this study. A total of 3 SAEs (Cholangitis, Breast abscess, and Diverticulitis) were reported in 2 subjects, but none were reported by the investigator or CSL as related to CSL324. Of the 3 SAEs, Breast abscess and Diverticulitis were under the SOC of Infections and infestations.

[0584] • Serious infections related to administrations of CSL324 were not observed. Overall, events under the SOC of Infections and infestations occurred in 21 subjects. Of these subjects, 4 subjects (1 HS, 3 PPP; all Cohort 2) had coinciding Neutropenia events, of which most were mild and 1 was moderate (resolved).

[0585] • Two subjects (5.1%) experienced 2 TEAEs (moderate Neutropenia and moderate Hypersensitivity) that led to permanent discontinuation of CSL324 and withdrawal from the study.

[0586] • With the exception of decreases in ANC, there were no clinically relevant trends in hematology parameters over time. No clinically relevant trends in biochemistry and urinalysis parameters were observed over time. Two clinically significant urinalysis abnormalities were observed for 1 subject with HS; both events resolved. No clinically relevant changes in vital signs or physical examination findings were observed over time.

[0587] • One subject had treatment-emergent antidrug antibodies at a low titer of 20 at 2 timepoints, which resolved on subsequent visits.

[0588] Conclusions

[0589] Overall, repeat doses of CSL324 were safe and well tolerated in subjects with HS and in subjects with PPP, where the majority of TEAEs observed were mild in severity. There were no deaths in this study. A total of 3 SAEs (Cholangitis, Breast abscess, and Diverticulitis) were reported in 2 subjects, but none were reported by the investigator or CSL as related to CSL324. The most common CSL324-related TEAEs were Fatigue and Neutropenia. The most common CSL324-related TEAEs in subjects with HS were Fatigue and Nausea; the most common CSL324-related TEAEs in subjects with PPP were Fatigue, Neutropenia, and Headache. The treatment-emergent AESIs reported were 4 events of moderate Neutropenia, all of which were related to CSL324 and resolved without treatment. Further, 1 event of moderate Pyrexia, which occurred in conjunction with a Grade 3 infection (nature and site of infection not specified), was reported in 1 subject with HS and reported as not related to administration of CSL324. One AESI of Neutropenia led to permanent discontinuation of CSL324 in 1 subject with HS. No serious or fatal AESIs occurred in the study. Serious infections related to administration of CSL324 were not observed. No clear correlation of infections to Neutropenia was observed.

[0590] The serum concentration and PK parameters of CSL324 were generally similar in subjects with HS and subjects with PPP. Following the first and last doses, mean Cmax increased in a dose-proportional manner; however, the increase in mean AUCiast was greater than dose proportional. Receptor occupancy (as measured by serum G-CSF concentrations) was not maintained throughout the dosing period in Cohort 1 but was maintained in some but not all subjects in Cohort 2. Overall, there was a dose-dependent decrease in mean ANC values consistent with the pharmacology of CSL324. The trend of ANC values after infusion of CSL324 was similar between Cohort 1 and Cohort 2. For Cohort 2, the extent of decrease in ANC values was greater than that for Cohort 1, and ANC values continued to stay suppressed until the next infusion unlike those observed for Cohort 1. Thus, with increasing dose, there was a clear dose-dependent increase in drug exposure leading to dose-dependent increase in PD activity measured by the PD markers serum G-CSF and ANC. Furthermore, in HS patients, there was a correlation between the decrease in ANC values and clinical outcome (HiSCR50).

[0591] Example 2: A Phase 1, Randomized, Double-blind, Placebo-controlled Study to Evaluate the Pharmacokinetics and Safety of Subcutaneous CSL324 in Healthy Japanese and White Subjects (CSL324_1003)

[0592] Objectives

[0593] Primary Objectives: 1. To characterize the pharmacokinetics (PK) of CSL324 in Japanese and White subjects following a single subcutaneous (SC) administration of CSL324.

[0594] 2. To compare the PK of CSL324 in Japanese and White subjects following a single SC administration of CSL324.

[0595] Secondary Objectives:

[0596] 1. Evaluate the safety and tolerability of a single SC dose of CSL324 in Japanese and White subjects.

[0597] 2. Further characterize the PK properties of CSL324 in healthy Japanese and White subjects.

[0598] 3. Investigate the immunogenicity of a single SC dose of CSL324.

[0599] Exploratory Objective: To assess the pharmacodynamic (PD) properties of CSL324 after a single SC dose of CSL324 in Japanese and White subjects.

[0600] Methodology

[0601] This was a single-site, randomized, double-blind, placebo -controlled study investigating the PK and safety of a single SC dose of CSL324 in healthy adult Japanese and White subjects.

[0602] This study consisted of a Screening Period from Days -28 to -2, an In-house Period from Days -1 to 5, and a Follow-up Period starting on Day 6 and ending at the End of Study Visit on Day 56. Healthy volunteers were screened and eligible subjects were admitted to study site on Day -1 and randomized. On Day 1, subjects received a single SC injection of CSL324 or placebo. Blood samples for various assessments, including PK, hematology, and granulocyte colony -stimulating factor (G-CSF), were collected before and at 30 minutes, 1 hour, 2 hours, 4 hours, and 12 hours after injection on Day 1 and then daily until Day 5. Subjects were discharged from the study site on the afternoon of Day 5 and were required to attend 7 follow-up visits and the End of Study Visit for further blood sampling and safety assessments.

[0603] The study consisted of 2 dose cohorts (Cohort 1: 0.3 mg / kg CSL324 and Cohort 2: 0.8 mg / kg CSL324). The study began with Cohort 1 and proceeded to Cohort 2 after all subjects in Cohort 1 were administered the investigational product (IP) and their safety data up to Day 14 had been reviewed by the Safety Review Committee (SRC). Each dose cohort consisted of 2 subcohorts: 1 subcohort of Japanese subjects (ie, Cohorts 1A and 2A) and 1 subcohort of White subjects (ie, Cohorts IB and 2B). Each subcohort comprised 8 subjects who were randomized to either CSL324 (6 subjects) or placebo (2 subjects) on Day -1. The first 2 subjects enrolled in Cohort 1A and IB (sentinel subjects) were randomized (1 CSL324 : 1 placebo), administered the IP, and monitored for 4 days. Safety data collected during the 4-day monitoring period was assessed by the Investigator and the CSL Medical Monitor for clinically significant safety issues. None were identified, and the next 6 subjects were randomized (5 CSL324 : 1 placebo) and administered IP.

[0604] Safety, Pharmacokinetic, and Pharmacodynamic Assessments

[0605] Safety assessments included treatment-emergent adverse events (TEAEs) (including AEs, of special interest [AESIs], serious AEs [SAEs], and AEs localized to the administration site), vital signs, 12-lead electrocardiogram (ECG), clinical laboratory tests (hematology, biochemistry, and urinalysis), and presence of anti-CSL324 antibodies in serum.

[0606] Blood samples were collected to assess the concentration of CSL324 in serum and to derive PK parameters. Blood samples were also collected to assess serial absolute neutrophil count (ANC) in whole blood (as part of the hematology safety assessment) and serum G-CSF concentration to assess target engagement.

[0607] Number of Subjects

[0608] Planned: minimum of 32 and maximum of approximately 48.

[0609] Actual: 32 (Cohort 1: 16; Cohort 2: 16).

[0610] Diagnosis and Main Criteria for Inclusion

[0611] Healthy male or female subjects, between 20 and 55 years of age, with a body mass index range of 18 to 32.0 kg / m2, and weight between 45 kg and 100 kg, of Japanese or White race, who provided written informed consent were included in the study. Female subjects were not pregnant, breastfeeding, or planning to become pregnant during the study or within 30 days after the End of Study Visit and were willing to use highly effective forms of contraception for the duration of the study. Male subjects with partners of childbearing potential were not planning pregnancy during the study or within 30 days after the End of Study Visit, and were willing to use highly effective forms of contraception for the duration of the study.

[0612] Subjects were excluded if they had sustained and / or symptomatic hypotension, a mean QT interval corrected using Fridericia's formula > 450 ms for male subjects or > 470 ms for female subjects, history of myeloproliferative or lymphoproliferative disease, history of active tuberculosis, concurrent diagnosis of malignancy or history of malignancy, clinically significant abnormalities on physical examination, vital signs, or laboratory assessments, or neutropenia (defined as ANC < 2.0 x 109 / L), evidence of hepatobiliary disease, immunosuppressive conditions, were currently taking immunosuppressive or immunomodulative therapy, had previous or current use of a biologic agent, including any investigational biologic agent, previous, current, or prospective use of any other prohibited therapy, clinical signs of active infection and / or fever > 38°C at Screening, history of chronic or recurrent infections, been hospitalized or received intravenous antibiotics for an infection within 2 months before Screening, confirmed HIV infection, active hepatitis B viral infection, hepatitis C viral infection, known or suspected hypersensitivity to CSL324 or to any excipients of CSL324, history of chronic alcohol or drug abuse within previous 1 year or a positive drug screen result and / or alcohol breath test at Screening, or were smokers, or donated or lost > 500 mL of blood or plasma within the 3 months before Day -1.

[0613] Investigational Product, Dose and Mode of Administration, Batch Number(s)

[0614] CSL324 was provided as a sterile solution for injection containing 120 mg / mL of CSL324 in a 2 mL vial. Subjects were administered a single SC injection of CSL324 on Day 1:

[0615] • Cohort 1 : 0.3 mg / kg CSL324.

[0616] • Cohort 2: 0.8 mg / kg CSL324.

[0617] The volume administered was determined by the subject’s dose cohort and weight on Day -1.

[0618] Placebo (formulation buffer for injection including all components of CSL324 except for the active ingredient) was provided as a sterile solution for injection in a 2 mL vial. Subjects were administered a single SC injection of placebo on Day 1. The volume administered was determined by the subject’s dose cohort and weight on Day -1.

[0619] Batch numbers for CSL324: 27212AC49A; 27212AC49B; 27212AC49C. Batch numbers for placebo: 27213AC43A; 27213AC43B; 27213AC43C.

[0620] Duration of Treatment

[0621] Subjects received CSL324 or placebo as a single dose on Day 1 and were followed up until Day 56.

[0622] Criteria for Evaluation

[0623] Primary Endpoints: • Maximum concentration (Cmax), area under the concentration-time curve from time 0 extrapolated to time infinity (AUCo-inf), and area under the concentration-time curve from time 0 to the last measurable concentration (AUCo-iast): oMean (standard deviation [SD]) and geometric mean (geometric % coefficient of variation [CV]). o90% confidence interval (CI) for the ratio of geometric means using White subjects as the reference.

[0624] Secondary Endpoints:

[0625] • TEAEs up to Day 56.

[0626] • AEs localized to the administration site up to Day 7.

[0627] • PK parameters: oTime to reach C max (T max). oTerminal half-life (ti / 2). o Apparent clearance (CL / F). oApparent volume of distribution (Vz / F).

[0628] • Presence of anti-CSL324 antibodies in serum on Day 1 before IP administration and on Days 28 and 56 after IP administration.

[0629] Statistical Methods

[0630] Analysis Populations

[0631] The Enrolled Analysis Set comprised all subjects who provided written informed consent and who were eligible for inclusion in the study at Screening. Screening failures were not included in the Enrolled Analysis Set.

[0632] The Safety Analysis Set comprised all subjects in the Enrolled Analysis Set who received at least 1 partial dose of IP.

[0633] The PK Analysis Set comprised all subjects in the Safety Analysis Set who were administered CSL324 and had at least 1 quantifiable concentration of CSL324 after administration.

[0634] The PD Analysis Set comprised all subjects in the Safety Analysis Set who had PD data from before administration and from at least 1 time point after administration of IP.

[0635] General Considerations

[0636] Continuous variables were summarized in terms of the number of observations, mean, standard deviation, minimum, first quartile (QI), median, third quartile (Q3), and maximum. Other descriptive statistics were reported when appropriate, eg, other quantiles, %CV (CV in percent was defined as CV = 100* SD / mean), geometric %CV (geometric CV in percent was defined as geometric CV = 100* square root of [{antilog of variance for log transformed data} - 1], and 90% CI. Categorical variables were summarized using frequency counts and percentages. As appropriate, a CI for a parameter was calculated by respective Statistical Analysis System procedure which had also calculated the designated parameter.

[0637] Pharmacokinetic Analyses

[0638] CSL324 PK parameters were natural log-transformed, and 90% Cis for the mean differences in Cmax, AUCo-inf, and AUCo-iast (in the log scale) between Japanese subjects and White subjects receiving the same dose of CSL324 were constructed based on a t distribution using pooled (across Japanese and White) estimate of variance.

[0639] The Cis were then exponentiated to form 90% Cis for the ratios of the geometric means in the original scale.

[0640] Pharmacodynamic Analyses (Biomarker Analyses)

[0641] Biomarker parameters were summarized by race, cohort, and treatment group for the PD Analysis Set by descriptive statistics with parameter of n, arithmetic mean, SD, %CV, minimum, QI, median, Q3, maximum, geometric mean, and geometric %CV.

[0642] Individual plot of concentration-time profiles and mean (± SD) profiles for ANC and G CSF were plotted using actual elapsed time and nominal (planned) time, respectively.

[0643] PK / PD Analyses

[0644] Pearson and Spearman correlation analyses were explored, where data permitted, and have been applied to PK parameters to explore the relationship between different components, especially for the changes associated with CSL324 administration. The PK and PD parameters (biomarkers) included Cmax versus ANC at the maximum (post- vs. predose) change from baseline and Cmax versus G-CSF at the maximum (post- vs. predose) change from baseline.

[0645] The correlation analyses were applied to each pair for all doses, whenever data permitted. If not specified otherwise, PD Analysis Set was used for the correlation analyses.

[0646] Results

[0647] Subject Disposition A total of 67 subjects provided informed consent and were screened, of which 32 subjects were enrolled in this study. Thirty-five subjects did not meet the eligibility criteria and were not enrolled in the study.

[0648] A total of 32 subjects were randomized into the study; 24 subjects received CSL324 and 8 subjects received placebo. Both Cohort 1 and Cohort 2 of the study included 16 subjects (8 Japanese and 8 White) each; of which 12 subjects (6 Japanese and 6 White) received CSL324 and 4 subjects (2 Japanese and 2 White) received placebo.

[0649] Overall, 31 subjects (96.9%) completed the study. One subject (placebo-treated) in Cohort 2B was discontinued due to Other reasons.

[0650] There were no major differences in demographic characteristics between subjects in Cohort 1 and Cohort 2. In each dose cohort, Japanese subjects had similar body weight (< 15% median difference) to White subjects.

[0651] Pharmacokinetic s

[0652] The serum CSL324 concentrations in Cohort 1 (0.3 mg / kg) were variable (%CV > 60% at all time points) across all subjects, with one 1 Japanese subject (Subject 0360111-0004) having no measurable concentrations at any time point and the majority of subjects having below limit of quantification (BLQ) values by Day 14.

[0653] Following a single SC administration of 0.3 mg / kg CSL324, median Tmax occurred at 72 hours in both Japanese and White subjects. Both Cmax, which had a mean of 1.474 pg / mL for White subjects and 1.157 pg / mL for Japanese subjects, and AUCo- iast, which had a mean of 240.347 h*pg / mL for White subjects and 184.994 h*pg / mL for Japanese subjects, showed similar variability between White and Japanese subjects (%CV ranging from 63.6% to 71.3%).

[0654] The serum CSL324 concentrations in Cohort 2 (0.8 mg / kg) were less variable than those seen in Cohort 1 (%CV < 50% at most time points after Day 1) across all subjects. Most subjects had BLQ values by Day 28.

[0655] Following a single SC administration of 0.8 mg / kg CSL324, median Tmax occurred at 157 hours in Japanese subjects and at 121 hours in White subjects. Compared with Japanese subjects, White subjects showed lower variability for Cmax (%CV of 17% vs. 45%) and AUCo-iast (6% vs. 48%).

[0656] With an increase in dose of 2.7-fold from 0.3 mg / kg to 0.8 mg / kg, mean Cmax and AUCo-iast increased 4.0-fold and 7.7-fold, respectively, in Japanese subjects and 3.5-fold and 7.3-fold, respectively, in White subjects. The AUCo-inf was calculable in only 3 of the 6 Japanese subjects in both 0.3 mg / kg and 0.8 mg / kg cohorts, compared to 5 of 6 White subjects in the 0.3 mg / kg cohort and 6 of 6 in the 0.8 mg / kg cohort, and should be interpreted with caution.

[0657] While there was overlap in the ranges of Cmax and AUCo-iast, Japanese subjects were in the slightly lower range in the 0.3 mg / kg cohort. However, the large variability observed precludes a definitive comparison of the PK parameters across White and Japanese subjects in the 0.3 mg / kg cohort. In the 0.8 mg / kg cohort, Japanese subjects showed similar Cmax and AUCo-iast estimates compared with White subjects, and values for White subjects fell completely within the range shown for Japanese subjects.

[0658] PK parameters between Japanese and White subjects expressed as geometric mean ratios and 90% Cis were compared. At the 0.3 mg / kg dose, the Cmax and AUCo-iast geometric mean values were 28% and 37% lower, respectively, in Japanese subjects compared to White subjects. At the 0.8 mg / kg dose, the Cmax and AUCo-iast geometric mean values were 16% and 26% lower, respectively, in Japanese subjects compared to White subjects. The observed variability in the data and the fact that the study was not powered to show bioequivalence mean these Cis are necessarily very wide, particularly in the 0.3 mg / kg cohort. The tighter Cis in the 0.8 mg / kg cohort and the fact that the Cis contain 1.0 provide some confidence that the PK parameters are similar between White and Japanese subjects.

[0659] Pharmacodynamic s

[0660] Japanese subjects experienced a similar drop in ANC values compared to White subjects in both dose cohorts. The largest decrease in mean ANC values was observed around Day 3 or 4 post CSL324 administration. While the extent of the decrease in ANC values was similar for both doses, the data suggests that the 0.8 mg / kg dose suppressed ANC values longer than the 0.3 mg / kg dose.

[0661] The mean serum G-CSF concentrations in Japanese subjects were comparable to White subjects in the 0.8 mg / kg dose cohort but were lower in the 0.3 mg / kg dose cohort. However, due to predose BLQ values in 5 of the 6 Japanese subjects in the 0.3 mg / kg dose cohort, these results should be interpreted with caution.

[0662] Safety

[0663] Overall, there were 60 TEAEs that occurred in 26 subjects (81.3%) receiving CSL324 or placebo during the study. Eight CSL324-treated subjects (66.7%) in Cohort 1, 12 CSL324-treated subjects (100.0%) in Cohort 2, and 6 placebo -treated subjects (75.0%) experienced at least 1 TEAE. The most common TEAE in the study was neutropenia (24 events occurring in 17 subjects [53.1%], all of whom received CSL324). All neutropenia events were transient, resolved or were resolving without treatment, and could be explained by the mechanism of action of CSL324. The other TEAEs that occurred in > 1 subject were headache (7 subjects [21.9%]), fatigue (3 subjects [9.4%]), and vessel puncture site bruise (2 subjects [6.3%]).

[0664] In Cohort 1, 5 White CSL324-treated subjects (83.3%) experienced 16 TEAEs and 3 Japanese CSL324-treated subjects (50.0%) experienced 10 TEAEs. In Cohort 2, 6 White CSL324-treated subjects (100.0%) experienced 14 TEAEs and 6 Japanese CSL324-treated subjects (100.0%) experienced 12 TEAEs.

[0665] Overall, 29 TEAEs that occurred in 18 subjects (56.3%) were related to CSL324. In Cohort 1, 7 subjects (58.3%) (4 White and 3 Japanese) experienced 12 TEAEs related to CSL324. In Cohort 2, 11 subjects (91.7%) (6 White and 5 Japanese) experienced 17 TEAEs related to CSL324.

[0666] The majority of TEAEs in the study were mild in severity. Four CSL324-treated subjects (33.3%) (2 White and 2 Japanese) in Cohort 1, 7 CSL324-treated subjects (58.3%) (3 White and 4 Japanese) in Cohort 2, and 4 placebo -treated subjects (50.0%) experienced a TEAE that was moderate in severity. Moderate TEAEs included neutropenia (9 subjects, [28.1%]), headache (6 subjects, [18.8%]), and iron deficiency, dermatitis, thermal burn, tooth impacted, and vulvovaginal candidiasis (1 subject [3.1%] each). Nine subjects had moderate TEAEs that were related to CSL324 (all were neutropenia), all of which were transient and resolved or were resolving without treatment. Nine subjects used concomitant medication or treatment for moderate non CSL342-related TEAEs of dermatitis, headache, thermal burn, tooth impacted, and vulvovaginal candidiasis. No severe TEAEs occurred during the study.

[0667] Local administration site reactions were reported by 4 subjects in the study. These included vessel puncture site bruise, catheter site haematoma, and injection site bruising. All events were mild in severity, not related to the study drug, and resolved without treatment.

[0668] There was 1 AESI, Grade 3 neutropenia, that was experienced by 1 Japanese subject in Cohort 2. The baseline ANC for the subject was 1.9 x 109 / L, which reached the lowest level of 0.7 x 109 / L on Day 21, increased to > 1.0 x 109 / L on Day 24, and completely resolved by Day 70 without treatment, when the subject’s neutrophils were 1.8 x 109 / L. NO other AESIs occurred during the study.

[0669] No subjects experienced Grade 3 or 4 infection and there was no evidence of an increased susceptibility to infection noted in this study. There were no subjects with presence of anti-CSL324 antibodies in serum over the treatment period.

[0670] There were no deaths or SAEs in this study. No subject discontinued / withdrew from the study due to TEAEs.

[0671] Conclusions

[0672] While there was overlap in the ranges of Cmax and AUCo-iast, Japanese subjects were in the slightly lower range in the 0.3 mg / kg cohort. However, the large variability observed precludes a definitive comparison of the PK parameters across White and Japanese subjects in the 0.3 mg / kg cohort. In the 0.8 mg / kg cohort, Japanese subjects showed similar Cmax and AUCo-iast estimates compared with White subjects, and values for White subjects fell completely within the range shown for Japanese subjects. In conclusion, the PK of CSL324 in Japanese and White subjects were comparable following a single SC administration of 0.8 mg / kg CSL324. Japanese subjects experienced a similar drop in ANC values compared to White subjects in both dose cohorts. While the extent of the decrease in ANC values was similar for both CSL324 doses, the data suggests that the 0.8 mg / kg dose suppressed ANC values longer than the 0.3 mg / kg dose.

[0673] CSL324 was safe and well tolerated and the incidence of TEAEs, with the exception of neutropenia, was similar between CSL324-treated and placebo-treated subjects. The most common CSL324-related TEAE was neutropenia (40.0% of all TEAEs). All neutropenia events occurred in CSL324-treated subjects, could be explained by the mechanism of action of CSL324, were transient, and resolved without treatment. The incidence of neutropenia was similar between Japanese and White subjects in both dose cohorts. No immunogenicity or major local tolerability issues occurred in the study.

[0674] Example 3: G-CSF gene signature present in adalimumab-failed HS patients

[0675] A gene signature of G-CSF activity was derived using the transcriptomic data generated from peripheral samples collected from the first in human clinical trial with CSL324, the CSL324_1001 (ACTRN12616000846426), a single-dose ascending and repeated dose, randomized, double-blind, placebo-controlled study to assess the safety, pharmacokinetics and pharmacodynamics of CSL324 in healthy adult subjects.

[0676] The description of the generation of the blood transcriptome dataset was published in Gamell et al., 2023. The key features of this signature included being representative of G-CSF stimulation and compatibility with signature enrichment algorithms. The signature was derived from genes that were differentially expressed (adjusted p-value < 0.05 and log2 fold-change > |2|) at Day 3 (post-filgrastim / G-CSF) vs Pre-Treatment in the placebo group. This list was subsequently filtered to remove genes that are lowly expressed (~log2counts < 0). All genes with reduced expression post-G- CSF were subsequently removed to focus on upregulated genes only. Genes were then also filtered to only include protein-coding genes. The resulting gene signature, named the “In-house G-CSF gene signature”, contains a total of 239 genes and it is depicted in the heatmap in Figure 1.

[0677] G-CSF gene signature is elevated in skin from HS patients, and it remains elevated in adalimumab non-responders but not in responders

[0678] The gene expression of the G-CSF gene signature was evaluated in skin samples from patients with HS who received adalimumab. The gene expression data were sourced from the study conducted by Lowe et al., available under the reference GSE155176. Lowe et al's publication shared RNA sequencing data obtained from lesional skin of HS patients involved in the PIONEER studies. These studies, pivotal phase 3 multicentre trials, investigated the efficacy of adalimumab (40 mg weekly) for HS through doubleblind, placebo-controlled periods. Lowe et al's publication included gene expression data from both pre-treatment (n=14) and posttreatment (n=15) skin samples of HS patients receiving adalimumab, along with gene expression data from healthy skin samples (n=16).

[0679] The expression levels of genes in the G-CSF gene signature within lesional skin samples from the adalimumab trial were examined. This analysis was conducted through sample-level enrichment analysis using the R package GSVA (intps: / / gldnih,conVrx?ass&lo / GSV A) (Hanzelmann et al., 2013). The GSVA results provided an enrichment score for each individual sample, where a positive score signifies increased expression of genes in the G-CSF gene signature, indicating activation of the G-CSF signalling pathway. The outcomes of this analysis are depicted in Figure 2.

[0680] The G-CSF gene signature exhibited enrichment in the lesional skin of HS patients (HS lesional_PRE) in comparison to skin obtained from healthy donors (Normal). Following adalimumab treatment, the signature was no longer enriched in responders (depicted by black dots, HS lesional_POST) but remained elevated in non- responders (illustrated by white dots, HS lesional_POST). This suggests that treatment with CSL324 may be beneficial in adalimumab non-responders, providing a compelling rationale to further investigations to assess the efficacy of CSL324 in individuals who do not respond to anti-TNF therapy. Thl7 / IL-17 gene-sets did not segregate adalimumab non-responders from responders

[0681] Activation of Thl7 / IL-17 pathways was also assessed in the same samples. Biologic drugs that specifically target IL-17A, such as secukinumab and ixekizumab, are currently being studied for their potential efficacy in treating HS.

[0682] In this instance, as the inventors lack in-house generated activated Thl7 cells or IL- 17 gene signatures, the inventors utilized public gene sets to monitor Th 17 and IL- 17 pathways, drawing from the Molecular Signatures Database (MSigDB), a repository of gene sets commonly employed in the analysis of gene expression data. MSigDB serves as a valuable resource for researchers in genomics and bioinformatics, aiding in the interpretation and analysis of high-throughput gene expression data within the context of biological pathways, functions, and processes (https: / / www.gsea- msigdb . org / g sea / msigdb / ) .

[0683] The inventors identified 19 gene sets in the MSigDB database (C2 and C5_GO:BP collections) that serve as indicators of Thl7 cell activation and / or IL- 17 signalling. GSVA analysis was conducted to evaluate the enrichment of these gene sets in the transcriptomic data obtained from skin samples in the PIONEER studies. Figure 3 illustrates the results obtained with three representative gene sets. The data suggests an elevation in Thl7 / IL-17 pathways in HS lesional skin, aligning with previous publications and expectations. However, unlike the G-CSF gene signature, the enrichment of Thl7 / IL-17 gene sets do not differentiate between adalimumab responders and non-responders in the skin samples collected post-adalimumab administration (HS lesional_POST).

[0684] Top disease-associated pathways enriched in HS lesions do not segregate adalimumab nonresponders from responders

[0685] Other key processes altered in the HS samples assessed to determine if they were differentially affected post-treatment in adalimumab responders vs non-responders, as it was the case for G-CSF signalling. The first step was to identify the top pathways altered in the HS skin samples from the PIONEER individuals. For that, the gene expression profile of the skin lesions from HS patients pre-adalimumab treatment vs the gene expression profile of the skin samples from healthy individuals was compared by performing fast ranked Gene Set Enrichment Analysis (fGSEA) using the R package ClusterProfiler (Yu et al., 2012). fGSEA was used to assess if certain groups of genes that work together in specific functions or pathways were more or less active. Figure 4 shows the top 10 pathways identified as differential regulated in skin samples from HS patients compared to healthy skin. The pathways were ranked by magnitude of activation difference between HS and normal skin (delta). The size of the dot corresponds to the p-value associated with that function or pathway. A larger dot indicates a smaller p-value, which in statistical terms suggests that the observed enrichment of genes in that particular function or pathway is less likely to be due to random chance. Thus, larger dots represent more significant or meaningful biological associations. As expected, the top altered pathways in skin from HS individuals are related to inflammatory processes.

[0686] The enrichment score of the top 3 activated pathways identified in the HS patients in the skin samples obtained post-adalimumab treatment were assessed. As shown in Figure 5, all three gene sets were positively enriched in the HS skin compared with healthy skin (Normal), but unlike the G-CSF gene signature, these gene sets do not separate adalimumab responders from non-re spenders in the skin samples taken post- adalimumab administration. This suggests that other pathogenic mechanisms in HS do not differ in adalimumab non-responders from responders.

[0687] Altogether, this data provides strong scientific rationale supporting the potential for CSL324 to have clinical benefit in anti-TNF non-responders.

[0688] Example 4: Evaluation of the Efficacy and Safety of Anumigilimab in Patients with Hidradenitis Suppurativa Who Have Had an Inadequate Response to Anti-TNF Therapy

[0689] The efficacy of anumigilimab administered to subjects with moderate to severe HS (i.e., IHS4 > 4) who have had an inadequate response to at least 12 weeks of treatment with an anti-TNF therapy is assessed.

[0690] The study will last for approximately 44 weeks and will consist of a 28-day Screening Period, a 16-week placebo-controlled Treatment Period A, a 16-week Treatment Period B, and an 8-week Follow-up Period.

[0691] Eligible subjects will be randomized in a ratio of 2: 1 to the anumigilimab or placebo arms by means of an interactive response technology (IRT) system using the baseline IHS4 score as the stratification factor. Subjects will receive anumigilimab or placebo for 16 weeks.

[0692] Assessments of efficacy, safety, PK, PD / disease biomarkers, and immunogenicity; and collection of samples for research and RNA sequencing will be performed. Efficacy assessments will include assessment of HS lesions by a blinded examiner, algorithmic derivatives (IHS4, > 50% reduction from baseline in the total abscess and nodule count with no increase in abscess or draining tunnel count [HiSCR50], > 75% reduction from baseline in the total abscess and nodule count with no increase in abscess or draining tunnel count [HiSCR75], modified Hidradenitis Suppurativa Clinical Response [mHiSCR], and > 55% decrease from baseline in International Hidradenitis Suppurativa Severity Score System score [IHS4-55]), and patient-reported outcomes (Dermatology Life Quality Index [DLQI], pain numerical rating scale [NRS], and Hidradenitis Suppurativa Quality of Life [HiSQOL]). A blinded examiner, who will not have access to the subjects’ ANC during the study, will perform all efficacy assessments requiring clinical judgment. Recording of concomitant therapies and monitoring for AEs will be done for the duration of subject participation in Treatment Period A.

[0693] For subjects in selected study sites, skin biopsies will be taken on Day 1 (prior to IP administration), on Week 7 (Day 43; prior to IP administration), and at the end of Treatment Period A (Week 16 [Day 112]). Gene expression and immunohistological analysis may be performed to investigate treatment response to anumigilimab, HS disease activity and changes in neutrophilic inflammation, and pathway related to HS disease activity (e.g., TNF signaling, IL- 17 signaling and the G-CSF gene signature).

[0694] The efficacy of anumigilimab will be measured by assessing the number of abscesses, nodules, and draining tunnels (fistulae or sinus tracts) to derive the IHS4 and other response criteria (HiSCR50, HiSCR75, Modified HiSCR, and IHS4-55). An abscess is a tender but fluctuating mass, > 10 mm in diameter, and is surrounded by an erythematous area; the middle of an abscess contains pus. A nodule (inflammatory nodule) is a raised, 3- dimensional, round, infiltrated lesion, > 10 mm in diameter. A draining tunnel is a raised, tender but fluctuating longitudinal mass of variable length and depth, ending at the skin surface [Lipsker et al., 2016, 232: 137]. A draining tunnel will have drainage expressed at rest or with compression of surrounding structures [Frew et al., 2021, 157:449]. Each subject should, as much as possible, have the number of abscesses, nodules, and draining tunnels counted and the severity assessed by the same blinded assessor (as described in the section entitled “Blinded Examiner” above) during the study. Additional efficacy assessments include the patient-reported outcomes of DLQI, HiSQOL, and pain scores, as well as quantification of abscess and nodule flares. A flare is defined as at least a 25% increase in abscess and nodule count with a minimum increase of 2 relative to baseline. The efficacy of anumigilimab will also be assessed by determining the International Hidradenitis Suppurativa Severity Score System (IHS4). The IHS4 is calculated using the following formula: (number of nodules multiplied by 1) + (number of abscesses multiplied by 2) + (number of draining tunnels multiplied by 4). An IHS4 of < 3 indicates mild HS, 4 to 10 indicates moderate HS, and > 11 indicates severe HS [Zouboulis et al., 2017, 177: 1401].

[0695] IHS4-55 is a recently reported dichotomous outcome measure that is defined as a > 55% decrease from baseline in IHS4 score [Tzellos et al., 2023, 37:395], The > 55% threshold was able to discriminate between patients treated with adalimumab or placebo and was associated with significant reduction in inflammatory lesions. IHS4-55 was validated using data from adalimumab- and placebo-treated subjects with HS in both the PIONEER II and the combined PIONEER I and PIONEER II studies [Tzellos et al., 2023, 37:395],

[0696] The efficacy of anumigilimab will also be assessed by determining the Hidradenitis Suppurative Clinical Response (HiSCR). The HiSCR is defined as a > 50% reduction from baseline in the total abscess and nodule count with no increase in abscess or draining tunnel count. HiSCR has been used in several phase 2 studies of HS [Kanni et al., 2018, 138:795; Kimball et al., 2016a, 30:989; Tzanetakou et al., 2016, 152:52] and phase 3 studies (PIONEER I and II, SUNSHINE, and SUNRISE) [Kimball et al., 2016b, 375:422; Kimball et al., 2023, 401 :747], Likewise, the HiSCR75 is defined as a > 75% reduction from baseline in the total abscess and nodule count with no increase in abscess or draining tunnel count [Glatt et al., 2021, 157: 1279].

[0697] Modified HiSCR is another response criteria recently reported by InflaRX (InflaRX Controlling Inflammation. R&D Event: Vilobelimab and hidradenitis suppurativa. InflaRX; February 3, 2022) and is defined as a 50% reduction from baseline in the sum of abscess, nodule, and draining tunnel count plus draining tunnels reduced > 50% from baseline.

[0698] Patient reported outcomes including the dermatology life quality index (DLQI), pain numerical rating scale (NRS) and Hidradenitis Suppurativa Quality of Life (HiSQOL) will also be used to assess efficacy of anumigilimab treatment.

Claims

CLAIMS1. A method of treating or delaying progression or reducing or inhibiting or hindering development of a neutrophil-associated condition in a subject, wherein the subject has had an inadequate clinical response to an anti-tumour necrosis factor (TNF) therapy for the neutrophil-associated condition, the method comprising administering to the subject a compound that inhibits granulocyte colony stimulating factor (G-CSF) signaling.

2. The method of claim 1, wherein the neutrophil-associated condition is selected from the group consisting of hidradenitis suppurativa (HS); palmoplantar pustulosis (PPP); pyoderma gangrenosum; Sweet syndrome; pyoderma gangrenosum, acne, and hidradenitis suppurativa (PASH); psoriasis; rheumatoid arthritis; ankylosing spondylitis; psoriatic arthritis; Crohn’s disease; juvenile idiopathic arthritis; ulcerative colitis and combinations thereof.

3. The method of claim 1 or 2, wherein the neutrophil- associated condition is a neutrophil-associated autoimmune condition.

4. A method of treating or delaying progression or reducing or inhibiting or hindering development of a neutrophil-associated skin condition in a subject, wherein the subject has had an inadequate clinical response to an anti-TNF therapy for the neutrophil-associated skin condition, the method comprising administering to the subject a compound that inhibits granulocyte colony stimulating factor (G-CSF) signaling.

5. The method of claim 4, wherein the neutrophil-associated skin condition is selected from the group consisting of hidradenitis suppurativa (HS); palmoplantar pustulosis (PPP); pyoderma gangrenosum; Sweet syndrome; pyoderma gangrenosum, acne, hidradenitis suppurativa (PASH); psoriasis and combinations thereof.

6. The method of claims 4 or 5, wherein the neutrophil-associated skin condition is HS.

7. The method of claim 5 or 6, wherein the HS is moderate to severe HS.

8. The method of any one of claims 5 to 7, wherein the HS is moderate to severe HS and the subject is an adolescent 12 years of age or older.

9. The method of any one of claims 1 to 8, wherein the subject has had an inadequate clinical response to at least 12 weeks of treatment with the anti-TNF therapy.

10. The method of any one of claims 1 to 9, wherein the anti-TNF therapy is an anti- TNFa antibody or a soluble TNF receptor-fusion protein.

11. The method of claim 10, wherein the anti-TNF therapy is adalimumab, infliximab, certolizumab, golimumab or etanercept.

12. The method of any one of claims 4 to 11, wherein the subject has had an inadequate response, or demonstrated intolerance to, or had a contraindication to, at least 3 months of oral antibiotics for the treatment of the neutrophil-associated skin condition.

13. The method of any one of claims 1 to 12, wherein administration of the compound that inhibits G-CSF signaling:(a) does not induce neutropenia; or(b) does not cause sustained grade 3 or grade 4 neutropenia in the subject for greater than seven consecutive days; or(c) does not induce grade 4 neutropenia; or(d) induces grade 2 or grade 3 neutropenia for two consecutive days or less; or(e) does not induce neutropenia for more than 2 consecutive days or more than 1 day.

14. The method of claim 13, wherein the neutropenia is not associated with a fever.

15. The method of any one of claims 1 to 14, wherein the compound that inhibits G- CSF signaling binds to G-CSF or G-CSF receptor (G-CSFR).

16. The method of claim 15, wherein the compound that inhibits G-CSF signaling is a protein comprising an antibody variable region that binds to or specifically binds to G- CSF or G-CSFR and neutralizes G-CSF signaling.

17. The method of claim 16, wherein the compound that inhibits G-CSF signaling is a protein comprising a Fv.

18. The method of claim 17, wherein the protein comprises:(i) a single chain Fv fragment (scFv);(ii) a dimeric scFv (di-scFv);(iii) a diabody;(iv) a triabody;(v) a tetrabody;(vi) a Fab;(vii) a F(ab’)2;(viii) a Fv;(ix) one of (i) to (ix) linked to a constant region of an antibody, Fc or a heavy chain constant domain (CH) 2 and / or CH3;(x) one of (i) to (ix) linked to albumin or a functional fragment or variants thereof or a protein that binds to albumin; or(xi) an antibody.

19. The method of any one of claims 16 to 18, wherein the protein comprises an antibody variable region that binds to or specifically binds to G-CSFR and competitively inhibits the binding of antibody C1.2G comprising a heavy chain variable region (VH) comprising a sequence set forth in SEQ ID NO: 4 and a light chain variable region (VL) comprising a sequence set forth in SEQ ID NO: 5 to G-CSFR.

20. The method of any one of claims 16 to 19 wherein the protein binds to an epitope comprising residues within one or two or three or four regions selected from 111-115, 170-176, 218-234 and / or 286-300 of SEQ ID NO: 1.

21. The method of any one of claims 1 to 20, wherein the compound that inhibits G- CSF signaling is an antibody comprising:(i) a VH comprising an amino acid sequence set forth in SEQ ID NO: 4 and a VL comprising an amino acid sequence set forth in SEQ ID NO: 5;(ii) a VH comprising an amino acid sequence set forth in SEQ ID NO: 2 and a VL comprising an amino acid sequence set forth in SEQ ID NO: 3;(iii) a VH comprising three CDRs of a VH comprising an amino acid sequence set forth in SEQ ID NO: 4 and a VL comprising three CDRs of a VL comprising an amino acid sequence set forth in SEQ ID NO: 5;(iv) a VH comprising three CDRs of a VH comprising an amino acid sequence set forth in SEQ ID NO: 2 and a VL comprising three CDRs of a VL comprising an amino acid sequence set forth in SEQ ID NO: 3; or(v) a VH comprising:(a) a CDR1 comprising a sequence set forth in SEQ ID NO: 6;(b) a CDR2 comprising a sequence set forth in SEQ ID NO: 7; and(c) a CDR3 comprising a sequence set forth in SEQ ID NO: 8; and a VL comprising:(a) a CDR1 comprising a sequence set forth in SEQ ID NO: 9;(b) a CDR2 comprising a sequence set forth in SEQ ID NO: 10; and(c) a CDR3 comprising a sequence set forth in SEQ ID NO: 11.

22. The method of any one of claims 1 to 21, wherein the compound that inhibits G-CSF signaling is an antibody comprising:(i) a VH comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 21 and a VL comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 22;(ii) a VH comprising three CDRs of a VH comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 21 and a VL comprising three CDRs of a VL comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 22;(iii) a VH comprising:(a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 23;(b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 24; and(c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 25; and a VL comprising:(a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 26;(b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 27; and(c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQID NO: 28.

23. The method of any one of claims 1 to 22, wherein the compound that inhibitsG-CSF signaling is an antibody comprising:(i) a heavy chain comprising a sequence set forth in SEQ ID NO: 14 and a light chain comprising a sequence set forth in SEQ ID NO: 15; or(ii) a heavy chain comprising a sequence set forth in SEQ ID NO: 16 and a light chain comprising a sequence set forth in SEQ ID NO: 15.

Images