Methods for treating neutrophilic skin conditions

Abstract

The present disclosure relates to a method of treating a neutrophilic skin condition in a subject. The methods comprise subcutaneous administration of a fixed dose of an antibody that inhibits granulocyte colony stimulating factor (G-CSF) signaling to the subject.

Description

[0001] METHODS FOR TREATING NEUTROPHILIC SKIN CONDITIONS

[0002] RELATED APPLICATION DATA

[0003] The present application claims priority from United States Patent Application No. 63 / 633,892 filed 15 April 2024 entitled “Methods for Treating Neutrophilic Skin Conditions”, 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 methods of treating neutrophilic skin conditions in a subject.

[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 (eg, 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, such as in neutrophilic skin conditions.

[0010] One example of a neutrophilic skin condition is hidradenitis suppurativa (HS). HS is a chronic, recurrent, and painful inflammatory disease that affects the apocrine gland-bearing skin and is characterized by abscesses, nodules, and draining tunnels located on the skin under the breast and of the underarm, thigh, groin, and / or buttocks. The reported prevalence of HS ranges from 0.1% to 0.3% and about 75% of these patients are considered to have moderate to severe disease.

[0011] Current first-line treatment options for neutrophilic 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, adalimumab is an approved biologic therapy for the treatment of moderate to severe HS in the United States and European Union. Adalimumab was effective in approximately 40% to 60% of subjects, with loss of response in approximately 40% to 50% of subjects by Week 36 of treatment.

[0012] Accordingly, based on the foregoing the skilled person will recognise there remains a significant need for improved therapies for treating neutrophilic skin conditions.

[0013] SUMMARY OF INVENTION

[0014] In producing the present invention, the inventors sought to develop a convenient, safe and effective dosing regimen for treating neutrophilic skin conditions with an antibody that inhibits G-CSF signaling. In doing so, the inventors identified that such an antibody can be administered subcutaneously at a fixed dose, irrespective of the subject’s weight. In this regard, the inventors determined that a fixed dose of 25 mg or 50 mg can be administered subcutaneously every 13-15 days to provide a sufficient exposure to the antibody for effective treatment, while mitigating the risk of severe neutropenia. Such a dosage regimen is advantageous over previously reported intravenously administered weight-based doses, because subcutaneous administration means that the doses may be self-administered. In addition, fixed doses may be more cost-effective and convenient for both the manufacturer and patient.

[0015] Accordingly, the present disclosure provides a method of treating a neutrophilic skin condition in a human subject suffering from the neutrophilic skin condition, the method comprising administering subcutaneously to the subject a fixed dose of 25 mg or 50 mg of an antibody that inhibits granulocyte colony stimulating factor (G-CSF) signaling every 13-15 days.

[0016] The present disclosure also provides an antibody that inhibits granulocyte colony stimulating factor (G-CSF) signaling for use in a method of treating a neutrophilic skin condition in a human subject suffering from the neutrophilic skin condition, the method comprising administering subcutaneously to the subject a fixed dose of 25 mg or 50 mg of the antibody every 13-15 days.

[0017] The present disclosure also provides use of an antibody that inhibits granulocyte colony stimulating factor (G-CSF) signaling in the manufacture of a medicament for treating a neutrophilic skin condition in a human subject suffering from the neutrophilic skin condition, wherein a fixed dose of 25 mg or 50 mg of the antibody is administered subcutaneously to the subject every 13-15 days. Advantageously, the doses of antibody administered to the subject in accordance with the methods of the invention are fixed doses, i.e., not weight based (per kg) doses.

[0018] In some examples, the fixed dose is administered to the subject every 14 days.

[0019] In some examples, a fixed dose of 50 mg of the antibody is administered to the subject every 13-15 days. In some examples, a fixed dose of 25 mg of the antibody is administered to the subject every 13-15 days. In some examples, the antibody is administered to the subject for a total period of at least 16 weeks. In some examples, the antibody is administered to the subject for a total period of at least 32 weeks.

[0020] In some examples:

[0021] (i) the antibody is administered every 13-15 days during a first dosing period; and / or

[0022] (ii) the antibody is administered every 13-15 days during a second dosing period.

[0023] The “first dosing period" may be a “treatment period” or a “loading period” and the “second dosing period” may be a “maintenance period”. The second dosing period is subsequent to the first dosing period. The second dosing period may be optional in some examples of the methods of the invention.

[0024] The present disclosure also provides a method of treating a neutrophilic skin condition in a human subject suffering from the neutrophilic skin condition, the method comprising:

[0025] (i) administering subcutaneously to the subject a fixed dose of 25 mg or 50 mg of an antibody that inhibits granulocyte colony stimulating factor (G-CSF) signaling during a first dosing period; and

[0026] (ii) subsequently administering subcutaneously to the subject a fixed dose of 25 mg or 50 mg of the antibody that inhibits G-CSF during a second dosing period, wherein the antibody that inhibits G-CSF is administered every 13-15 days during the first dosing period and / or the second dosing period.

[0027] The present disclosure also provides an antibody that inhibits granulocyte colony stimulating factor (G-CSF) signaling for use in a method of treating a neutrophilic skin condition in a human subject suffering from the neutrophilic skin condition, the method comprising:

[0028] (i) administering subcutaneously to the subject a fixed dose of 25 mg or 50 mg of the antibody during a first dosing period; and

[0029] (ii) subsequently administering subcutaneously to the subject a fixed dose of 25 mg or 50 mg of the antibody during a second dosing period, wherein the antibody is administered every 13-15 days during the first dosing period and / or the second dosing period.

[0030] The present disclosure also provides use of an antibody that inhibits granulocyte colony stimulating factor (G-CSF) signaling in the manufacture of a medicament for treating a neutrophilic skin condition in a human subject suffering from the neutrophilic skin condition, wherein:

[0031] (i) a fixed dose of 25 mg or 50 mg of the antibody is administered subcutaneously to the subject during a first dosing period; and

[0032] (ii) a fixed dose of 25 mg or 50 mg of the antibody is subsequently administered subcutaneously to the subject during a second dosing period, wherein the antibody is administered every 13-15 days during the first dosing period and / or the second dosing period.

[0033] In some examples, a fixed dose of 50 mg of the antibody is administered every 13-15 days during the first period. In some examples, a fixed dose of 25 mg of the antibody is administered every 13-15 days during the first period.

[0034] In some examples, a fixed dose of 25 mg of the antibody is administered every 13-15 days during the second period. In some examples, a fixed dose of 50 mg of the antibody is administered every 13-15 days during the second period.

[0035] In some examples, a fixed dose of 50 mg of the antibody is administered every 13-15 days during the first period and a fixed dose of 25 mg of the antibody is administered every 13- 15 days during the second period.

[0036] In some examples, a fixed dose of 50 mg of the antibody is administered every 13-15 days during the first period and a fixed dose of 50 mg of the antibody is administered every 13- 15 days during the second period.

[0037] In some examples, a fixed dose of 25 mg of the antibody is administered every 13-15 days during the first period and a fixed dose of 25 mg of the antibody is administered every 13- 15 days during the second period.

[0038] In some examples, the antibody is administered every 14 days during the first dosing period and during the second dosing period.

[0039] In some examples, the first dosing period is in the range of 12 to 20 weeks. In some examples, the first dosing period is in the range of 14 to 18 weeks. In some examples, the first dosing period is in the range of 15 to 17 weeks. In some examples, the first dosing period is about 16 weeks. In some examples, the first dosing period is at least 16 weeks.

[0040] In some examples, the second dosing period is in the range of 12 to 20 weeks. In some examples, the second dosing period is in the range of 14 to 18 weeks. In some examples, the second dosing period is in the range of 15 to 17 weeks. In some examples, the second dosing period is about 16 weeks. In some examples, the second dosing period is at least 16 weeks.

[0041] In some examples, the antibody is administered every 13-15 days until the condition or symptoms of the condition are resolved or managed. In some examples, the antibody is administered for a total period of at least 16 weeks, at least 20 weeks, at least 24 weeks, at least 28 weeks, or at least 32 weeks. In some examples, the antibody is administered for a total period of at least 16 weeks. In some examples, the antibody is administered for a total period of at least 32 weeks.

[0042] In some examples, the antibody is administered to induce remission of a condition. In some examples, the antibody is administered to maintain remission of a condition.

[0043] In some examples, the subject has previously received treatment with an anti-tumour necrosis factor (TNF) therapy. For example, the anti-TNF therapy may be an anti-TNFa antibody, such as adalimumab.

[0044] In some examples, the subject has had an inadequate clinical response to the anti-TNF therapy. 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. Such subjects may also be described as having a neutrophilic skin condition which is “refractory to” the anti-TNF therapy. In some examples, the subject has had an inadequate clinical response to at least 12 weeks of treatment with the anti-TNF therapy. An inadequate clinical response may be assessed by a clinician after or during the anti-TNF therapy. For example, the inadequate clinical response may be an inadequate improvement in International Hidradenitis Suppurativa Severity Score System (IHS4), total abscess and nodule count, Dermatology Life Quality Index (DLQI), and / or Patient’s Global Assessment (PGA) of skin pain. For example, a subject who has had an inadequate clinical response to an anti-TNF therapy may be a subject who has an IHS4 score of equal to or greater than 4 after treatment with the anti-TNF therapy. An inadequate response may refer to a failure to achieve a Hidradentis Suppurativa Clinical Response (HiSCR) from the anti-TNF therapy. A HiSCR is defined as a 50% reduction in overall abscess and nodule count with no increase in abscess count and no increase in draining fistula count (also referred to herein as “HiSCR50”). Alternatively, a HiSCR may be a 75% reduction in overall abscess and nodule count with no increase in abscess count and no increase in draining fistula count (also referred to herein as “H1SCR75”).

[0045] In some examples, the inadequate clinical response to an anti-TNF therapy is a failure to achieve HiSCR50 after at least twelve weeks of treatment with the anti-TNF therapy. For example, the subject may be a subject who has failed to achieve HiSCR50 after at least twelve weeks of prior treatment with adalimumab.

[0046] As described herein, the inventors have surprisingly identified that subjects who do not adequately respond to anti-TNF therapy have a gene expression signature in their skin lesions which is indicative of G-CSF activity which persists after treatment with the anti-TNF therapy. Whereas the same G-CSF gene signature does not persist in subjects who respond to the anti- TNF therapy. These findings suggest that therapy with an antibody that inhibits G-CSF signaling may be particularly effective in subjects who do not respond adequately to anti-TNF therapy. In some examples, 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 neutrophilic skin condition. 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.

[0047] In some examples, administration of the antibody does not cause grade 3 neutropenia or grade 4 neutropenia (or severe neutropenia) in the subject for greater than three consecutive days. In some examples, administration of the antibody 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 some examples, administration of the antibody does not cause grade 3 neutropenia or grade 4 neutropenia (or severe neutropenia) in the subject for greater than seven consecutive days.

[0048] In some examples, administration of the antibody does not induce grade 4 neutropenia following a single administration. In some examples, administration of the antibody 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 some examples, administration of the antibody does not induce grade 4 neutropenia following at least three administrations.

[0049] In some examples, administration of the antibody induces grade 2 or grade 3 neutropenia for less than two consecutive days following a single administration. In some examples, administration of the antibody 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 some examples, administration of the antibody induces grade 2 or grade 3 neutropenia for less than two consecutive days following at least three administrations.

[0050] In some examples, administration of the antibody does not cause sustained grade 3 or grade 4 neutropenia in the subject for greater than seven consecutive days.

[0051] In some examples, administration of the antibody:

[0052] (i) does not induce grade 4 neutropenia; or

[0053] (ii) does not induce neutropenia or the antibody induces grade 2 or grade 3 neutropenia for two consecutive days or less; or

[0054] (iii) does not induce neutropenia for more than 2 consecutive days or more than 1 day.

[0055] In some examples, the neutropenia is not associated with a fever. In some examples, the neutropenia is resolved without treatment. In some examples, the neutropenia is not associated with an infection, e.g., a serious infection, such as a tuberculosis infection.

[0056] In some examples, the subject has an absolute neutrophil count (ANC) of > 2.0 x 109 / L prior to receiving treatment with the antibody that inhibits G-CSF signaling. In some examples, the subject does not have one or more of the following prior to receiving treatment with the antibody:

[0057] (i) a history of myeloproliferative disease;

[0058] (ii) malignancy within 5 years of receiving treatment with the antibody;

[0059] (iii) a clinically significant history of uncontrolled renal, hepatic (including currently active hepatitis B virus and / or hepatitis C virus), hematologic, endocrine, neurologic, pulmonary, psychiatric and / or cardiac disease;

[0060] (iv) a primary or secondary immunodeficiency;

[0061] (v) clinical signs of active infection and / or fever > 38°C during the 7 days before receiving treatment with the antibody;

[0062] (vi) a history of chronic, serious, or recurrent infections;

[0063] (vii) has been hospitalized or received intravenous antibiotic therapy for an infection within 2 months of receiving treatment with the antibody; and

[0064] (viii) has > 20 draining tunnels.

[0065] In some examples, the neutrophilic skin condition is a neutrophilic dermatosis or a neutrophilic skin lesion.

[0066] In some examples, the neutrophilic dermatosis is selected from the group consisting of hidradenitis suppurativa (HS); palmoplantar pustulosis (PPP); amicrobial pustulosis of the folds (APF); psoriasis; plaque psoriasis; CARD14-mediated pustular psoriasis (CAMPS); cryopyrin associated periodic syndromes (CAPS); deficiency of interleukin- 1 receptor (DIRA); deficiency of interleukin-36 receptor antagonist (DIRTA); palmoplantar pustulosis (PPP); pyogenic arthritis; pyoderma gangrenosum and acne (PAPA); pyoderma gangrenosum, acne, and hidradenitis suppurativa (PASH); pyoderma gangrenosum(PG); skin lesions of Behcet’s disease; Still’s disease; Sweet syndrome; subcorneal pustulosis (Sneddon-Wilkinson); pustular psoriasis; acute generalized exanthematic pustulosis; infantile acropustulosis; synovitis, acne, pustulosis; hyperostosis and osteitis (SAPHO) syndrome; bowel-associated dermatosisarthritis syndrome (BAD AS); neutrophilic dermatosis of the dorsal hands; neutrophilic eccrine hidradenitis; erythema elevatum diutinum; and Pyoderma gangrenosum.

[0067] In some examples, the neutrophilic dermatosis is hidradenitis suppurativa (HS), palmoplantar pustulosis (PPP) or psoriasis. In some examples, the neutrophilic dermatosis is hidradenitis suppurativa (HS) or palmoplantar pustulosis (PPP).

[0068] In some examples, the neutrophilic dermatosis is hidradenitis suppurativa (HS).

[0069] In some examples, the subject is suffering from moderate to severe HS. HS severity can be assessed using any method known in the art. In some examples, HS severity is determined according to the International Hidradenitis Suppurativa Severity Score System (IHS4; Zouboulis et al., Br J Dermatol. 2017, 177:1401). An IHS4 score 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. Thus, in some examples, the subject has an International Hidradenitis Suppurativa Severity Score System (IHS4) score of > 4. In some examples, the subject has a IHS4 score of > 11.

[0070] An exemplary method disclosed herein is a method of treating hidradenitis suppurativa (HS) in a subject, wherein the subject:

[0071] (i) is suffering from moderate to severe HS and / or has an International Hidradenitis Suppurativa Severity Score System (IHS4) score of > 4; and

[0072] (ii) has had an inadequate response to an anti-tumour necrosis factor (TNF) therapy, the method comprising:

[0073] (i) administering subcutaneously to the subject a fixed dose of 25 mg or 50 mg of an antibody that inhibits granulocyte colony stimulating factor (G-CSF) signaling during a first dosing period; and

[0074] (ii) subsequently administering subcutaneously to the subject a fixed dose of 25 mg or 50 mg of the antibody that inhibits G-CSF during a second dosing period, wherein the antibody that inhibits G-CSF is administered every 13-15 days during the first dosing period and / or the second dosing period.

[0075] Another exemplary method disclosed herein is a method of treating hidradenitis suppurativa (HS) in a subject, wherein the subject:

[0076] (i) is suffering from moderate to severe HS and / or has an International Hidradenitis Suppurativa Severity Score System (IHS4) score of > 4; and

[0077] (ii) has had an inadequate response to an anti-tumour necrosis factor (TNF) therapy, the method comprising:

[0078] (i) administering subcutaneously to the subject a fixed dose of 25 mg or 50 mg of an antibody that inhibits granulocyte colony stimulating factor (G-CSF) signaling during a first dosing period; and

[0079] (ii) subsequently administering subcutaneously to the subject a fixed dose of 25 mg or 50 mg of the antibody that inhibits G-CSF during a second dosing period, wherein the antibody that inhibits G-CSF is administered every 13-15 days during the first dosing period and the second dosing period, and wherein the first dosing period is at least 16 weeks.

[0080] In some examples, the subject has stable HS for at least 2 months (i.e., at least 2 months prior to treatment with a method disclosed herein). The term “stable HS” in this context refers to HS in which symptoms have persisted or progressed for at least 2 months prior to treatment with a method disclosed herein.

[0081] In some examples, the subject has had at least 1 draining tunnel prior to receiving treatment. A draining tunnel, also known as a sinus tract or fistula, is a raised, tender but fluctuating longitudinal mass of variable length and depth, ending at the skin surface and which can leak at rest or under with compression of surrounding structures. Subjects with more severe disease tend to experience higher numbers of draining tunnels.

[0082] In some examples, the antibody that inhibits G-CSF signaling is administered in an amount sufficient to reduce or prevent inflammation.

[0083] In some examples, the antibody that inhibits G-CSF is administered in an amount sufficient to reduce or prevent an increase in the subject’s IHS4 score. In some examples, administration of the antibody provides a > 55% decrease from baseline in IHS4 score (e.g., following 16 weeks of treatment).

[0084] In some examples, an absolute change from baseline in IHS4 is achieved following 16 weeks treatment. In some examples, the absolute change is a reduction in IHS4 score. For example, the absolute change from baseline in IHS4 may be a reduction in IHS4 score by at least 1, at least 2, at least 3, at least 4 or at least 5. In some examples, the subject’s IHS4 score is reduced by at least 2, at least 4, at least 6, at least 8 or at least 10. In some examples, the subject’s IHS4 score is reduced by at least 6, at least 8, at least 10, or at least 12. As the skilled person will understand, reference to a parameter at “baseline” refers to the value of the parameter prior to beginning treatment with a method disclosed herein.

[0085] In some examples, following 16 weeks of treatment the subject’s IHS4 score is

[0086] (i) reduced by at least 2, at least 4, at least 6, at least 8 or at least 10; and / or

[0087] (ii) reduced by at least 55%.

[0088] The present disclosure also provides a method of treating hidradenitis suppurativa (HS) in a subject, wherein the subject:

[0089] (i) is suffering from moderate to severe HS and / or has an International Hidradenitis Suppurativa Severity Score System (IHS4) score of > 4; and

[0090] (ii) has had an inadequate response to an anti-tumour necrosis factor (TNF) therapy, the method comprising administering subcutaneously to the subject a fixed dose of 25 mg or 50 mg of an antibody that inhibits granulocyte colony stimulating factor (G-CSF) signaling every 13-15 days, and wherein an absolute reduction in the IHS4 score is achieved following a first dosing period of at least 16 weeks.

[0091] The present disclosure also provides a method of reducing or preventing an increase in an International Hidradenitis Suppurativa Severity Score System (IHS4) score in a subject, wherein the subject is suffering from moderate to severe HS and / or has an IHS4 score of > 4, the method comprising administering subcutaneously to the subject a fixed dose of 25 mg or 50 mg of an antibody that inhibits granulocyte colony stimulating factor (G-CSF) signaling every 13-15 days, and wherein an absolute reduction in the IHS4 score is achieved following a first dosing period of at least 16 weeks. The method may further comprise subsequently administering subcutaneously to the subject a fixed dose of 25 mg or 50 mg of the antibody that inhibits G-CSF every 13-15 days during a second dosing period, wherein the reduction in the IHS4 score is maintained during the second dosing period.

[0092] The present disclosure also provides a method for maintaining a reduction of an International Hidradenitis Suppurativa Severity Score System (IHS4) score in a subject suffering from Hidradenitis Suppurativa (HS) and who has achieved an absolute reduction in IHS4 score from baseline during a first dosing period, the method comprising administering subcutaneously to the subject a fixed dose of 25 mg or 50 mg of the antibody that inhibits G- CSF every 13-15 days during a second dosing period.

[0093] As described herein above, in some examples, 50 mg of the antibody may be administered every 13-15 days during the first period. In some examples, 25 mg of the antibody is administered every 13-15 days during the first period. In some examples, 25 mg of the antibody is administered every 13-15 days during the second period. In some examples, 50 mg of the antibody is administered every 13-15 days during the second period. In some examples, 50 mg of the antibody is administered every 13-15 days during the first period and 25 mg of the antibody is administered every 13-15 days during the second period. In some examples, 50 mg of the antibody is administered every 13-15 days during the first period and 50 mg of the antibody is administered every 13-15 days during the second period. In some examples, 25 mg of the antibody is administered every 13-15 days during the first period and 25 mg of the antibody is administered every 13-15 days during the second period.

[0094] In some examples, the IHS4 score is reduced by > 55% (IHS4-55) at the end of a dosing period of at least 16 weeks.

[0095] In some examples, a HiSCR50 is achieved at the end of a dosing period of at least 16 weeks. In some examples, a HiSCR75 is achieved at the end of a dosing period of at least 16 weeks. In some examples, a modified HiSCR response is achieved at the end of a dosing period of at least 16 weeks.

[0096] In some examples, a reduction in a Dermatology Life Quality Index (DLQI) score from baseline is achieved at the end of a dosing period of at least 16 weeks. As is known in the art, the DLQI is a validated questionnaire consisting of ten questions that assess the effect of skin conditions on the quality of life of an affected patient (Finlay et al. Clin Exp Dermatol. 1994, 19:210).

[0097] In some examples, a reduction in the number of inflammatory lesions (AN counts) from baseline is achieved at the end of a dosing period of at least 16 weeks.

[0098] In some examples, a reduction in Patient’s Global Assessment of Skin Pain is achieved at the end of a dosing period of at least 16 weeks. Assessment of skin pain may be performed using a pain numerical rating scale (NRS). The pain NRS is a unidimensional measure of pain intensity in adults, including those with chronic pain associated with skin conditions (Kimball et al. 2016, 375:422). The reduction in the patient’s global assessment of skin pain (e.g., assessed using the pain NRS) may be: a) a reduction in pain at its worst and pain on average from baseline; and / or b) a reduction pain at its worst and pain on average over the previous 24 hours; and / or b) a reduction in pain at its worst and pain on average over the previous 7 days.

[0099] In some examples, one or more of the following is achieved in the subject at the end of a first dosing period of at least 16 weeks:

[0100] (i) a HiSCR50 or HiSCR75 or modified HiSCR response;

[0101] (ii) a reduction in a Dermatology Life Quality Index (DLQI) score from baseline;

[0102] (iii) a reduction in the number of inflammatory lesions (AN counts) from baseline; and

[0103] (iv) a reduction in Patient’s Global Assessment of Skin Pain including: a) pain at its worst and pain on average over the previous 24 hours and / or b) pain at its worst and pain on average over previous 7 days.

[0104] An exemplary method disclosed herein is a method of treating hidradenitis suppurativa (HS) in a subject, wherein the subject:

[0105] (i) is suffering from moderate to severe HS and / or has an International Hidradenitis Suppurativa Severity Score System (IHS4) score of > 4; and

[0106] (ii) has had an inadequate response to an anti-tumour necrosis factor (TNF) therapy, the method comprising administering subcutaneously to the subject a fixed dose of 25 mg or 50 mg of an antibody that inhibits granulocyte colony stimulating factor (G-CSF) signaling every 13-15 days, and wherein following a first dosing period of at least 16 weeks one or more of the following is achieved:

[0107] (i) at least a 55% reduction in the IHS4 score;

[0108] (ii) a hidradenitis suppurativa clinical response (HiSCR) 50 or HiSCR 75 or modified HiSCR response;

[0109] (iii) a reduction in a Dermatology Life Quality Index (DLQI) score from baseline;

[0110] (iv) a reduction in the number of inflammatory lesions (AN counts) from baseline; and

[0111] (v) a reduction in Patient’s Global Assessment of Skin Pain including: a) pain at its worst and pain on average from baseline; b) pain at its worst and pain on average over the previous 24 hours and / or c) pain at its worst and pain on average over previous 7 days.

[0112] In some examples, the reduction in Patient’s Global Assessment of Skin Pain is at least a 30% reduction in numeric rating scale (NRS) and at least 1-unit reduction from baseline in worst skin pain.

[0113] In some examples, administration of the antibody that inhibits G-CSF signaling results in a reduction in the subject’s ANC. As described herein, the inventors determined that a decrease in the subject’s ANC is correlated with clinical outcome in treatment of HS (see Example 1).

[0114] In some examples, administration of the antibody that inhibits G-CSF signaling results in an increase in the quantity of G-CSF in the subject’s blood.

[0115] In some examples, administration of the antibody that inhibits G-CSF signaling results in a reduction in the quantity of soluble CD177 (sCD177) in the subject’s blood.

[0116] In some examples, administration of the antibody that inhibits G-CSF signaling results in a reduction in the quantity of calprotectin in the subject’s blood.

[0117] In some examples, administration of the antibody that inhibits G-CSF signaling results in a reduction in the quantity of ECN-2 in the subject’s blood.

[0118] In some examples, administration of the antibody that inhibits G-CSF signaling results in an increase in the quantity of IE-8 in the subject’s blood.

[0119] In some examples, administration of the antibody that inhibits G-CSF signaling results in one or more or all of the following: a) a reduction in the subject’s ANC; b) an increase in the quantity of G-CSF in the subject’s blood; c) a reduction in the quantity of sCD177 in the subject’s blood; d) a reduction in the quantity of calprotectin in the subject’s blood; e) a reduction in the quantity of LCN-2 in the subject’s blood; and f) an increase in the quantity of IL-8 in the subject’s blood.

[0120] The reduction or increase in the above biomarkers can be assessed using any method known in the art, including those described herein. The reduction or increase may be assessed at a suitable time after administration of the antibody that inhibits G-CSF signaling, relative to an equivalent subject who has not been administered the antibody or relative to the level of the biomarker in the subject prior to administration. For example, the reduction or increase may be assessed about 24 to 72 hours after a dose of the antibody that inhibits G-CSF signaling, for example, about 24 to 72 hours after a first, a second, a third, a fourth, a fifth or further dose of the antibody. Alternatively, or additionally, the reduction or increase may be assessed after a dosing period of at least 4 weeks, at least 6 weeks, at least 8 weeks, at least 10 weeks, at least 12 weeks, at least 14 weeks or at least 16 weeks.

[0121] In some examples, the antibody that inhibits G-CSF signaling is an antibody binds to G- CSFR (the G-CSF receptor). Reference herein to an antibody that “binds to” G-CSFR provides literal support for an antibody that “binds specifically to” G-CSFR. Alternatively, the antibody that inhibits G-CSF signaling may be an antibody that binds to G-CSF. Reference herein to an antibody that “binds to” G-CSF provides literal support for an antibody that “binds specifically to” G-CSF. In some examples, the antibody comprises a heavy chain variable region (VH). In some examples, the antibody comprises a light chain variable region (VL). In some examples, the antibody comprises a VH and a VL. In some examples, the VH and a VL are in the same polypeptide chain. In another example, the VH and a VL are in separate polypeptide chains.

[0122] In some examples, the antibody 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 some examples, the antibody that inhibits G-CSF signaling comprises a Fv. The skilled artisan will understand that the antigen binding domain comprises the binding site of the antibody.

[0123] In some examples, the VH and the VL are in a single polypeptide chain. For example, the antibody may be or comprise:

[0124] (i) a single chain Fv fragment (scFv);

[0125] (ii) a dimeric scFv (di-scFv);

[0126] (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

[0127] (iv) one of (i) or (ii) linked to a protein that binds to an immune effector cell.

[0128] In some examples, the VL and VH are in separate polypeptide chains.

[0129] For example, the antibody may be or comprise:

[0130] (i) a diabody;

[0131] (ii) a triabody;

[0132] (iii) a tetrabody;

[0133] (iv) a Fab;

[0134] (v) a F(ab’)2;

[0135] (vi) a Fv; or

[0136] (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.

[0137] In some examples, the antibody comprises or is one of the following:

[0138] (i) a single chain Fv fragment (scFv);

[0139] (ii) a dimeric scFv (di-scFv);

[0140] (iii) a diabody;

[0141] (iv) a triabody;

[0142] (v) a tetrabody;

[0143] (vi) a Fab;

[0144] (vii) a F(ab’)2;

[0145] (viii) a Fv;

[0146] (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; or (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.

[0147] The antibody may be a conventional, i.e., a full length or whole, IgG antibody.

[0148] The foregoing proteins (described in the previous three lists) can also be referred to as antigen binding domains of antibodies.

[0149] In some examples, the antibody is a naked antibody. Exemplary antibodies are described in W02012171057, WO2018145206 and WO2022055334, all of which are incorporated herein by reference.

[0150] In some examples, the antibody binds to hG-CSFR expressed on the surface of a cell at an affinity of at least about 5 nM. In some examples, the antibody binds to hG-CSFR expressed on the surface of a cell at an affinity of at least about 4 nM. In some examples, the antibody binds to hG-CSFR expressed on the surface of a cell at an affinity of at least about 3 nM. In some examples, the antibody binds to hG-CSFR expressed on the surface of a cell at an affinity of at least about 2 nM. In some examples, the antibody binds to hG-CSFR expressed on the surface of a cell at an affinity of at least about 1 nM.

[0151] In some examples, the antibody inhibits G-CSF-induced proliferation of a BaF3 cell expressing hG-CSFR with an IC50 of at least about 5 nM. In some examples, the antibody inhibits G-CSF-induced proliferation of a BaF3 cell expressing hG-CSFR with an IC50 of at least about 4 nM. In some examples, the antibody inhibits G-CSF-induced proliferation of a BaF3 cell expressing hG-CSFR with an IC50 of at least about 3 nM. In some examples, the antibody inhibits G-CSF-induced proliferation of a BaF3 cell expressing hG-CSFR with an IC50 of at least about 2 nM. In some examples, the antibody inhibits G-CSF-induced proliferation of a BaF3 cell expressing hG-CSFR with an IC50 of at least about 1 nM. In some examples, the antibody inhibits G-CSF-induced proliferation of a BaF3 cell expressing hG- CSFR with an IC50 of at least about 0.5 nM.

[0152] In some examples, the antibody is chimeric, de-immunized, humanized, human or primatized. In some examples, the antibody is human.

[0153] In some examples, the antibody comprises an antigen binding domain 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.

[0154] In some examples, the antibody 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. In some examples, the antibody binds to an epitope comprising residues within all of the regions 111-115, 170-176, 218-234 and 286-300 of SEQ ID NO: 1. In some examples, the antibody binds to hG-CSFR expressed on the surface of a cell at an affinity of at least about 1 nM, wherein the antibody binds to an epitope comprising residues within all of the regions 111-115, 170-176, 218-234 and 286-300 of SEQ ID NO: 1.

[0155] In some examples, the antibody comprises 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.

[0156] In some examples, the antibody comprises 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.

[0157] In some examples, the antibody comprises 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.

[0158] In some examples, the antibody comprises 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.

[0159] In some examples, the CDRs are positioned according to the numbering system of Kabat.

[0160] In some examples, the antibody comprises a VH and a VL, wherein:

[0161] (i) the VH comprises: a. a CDR1 comprising a sequence set forth in amino acids 31-35 of SEQ ID NO: 2; b. a CDR2 comprising a sequence set forth in amino acids 50-65 of SEQ ID NO: 2; and c. a CDR3 comprising a sequence set forth in amino acids 99-107 of SEQ ID NO: 2; and

[0162] (ii)the VL comprises: a. a CDR1 comprising a sequence set forth in amino acids 24-34 of SEQ ID NO: 3; b. a CDR2 comprising a sequence set forth in amino acids 51-56 of SEQ ID NO: 3; and c. a CDR3 comprising a sequence set forth in amino acids 89-97 of SEQ ID NO: 3.

[0163] In some examples, the antibody comprises a VH and a VL, wherein:

[0164] (i) the VH comprises: a. a CDR1 comprising a sequence set forth in amino acids 31-35 of SEQ ID NO: 4; b. a CDR2 comprising a sequence set forth in amino acids 50-65 of SEQ ID NO: 4; and c. a CDR3 comprising a sequence set forth in amino acids 99-107 of SEQ ID NO: 4; and

[0165] (ii)the VL comprises: a. a CDR1 comprising a sequence set forth in amino acids 24-34 of SEQ ID NO: 5; b. a CDR2 comprising a sequence set forth in amino acids 51-56 of SEQ ID NO: 5; and c. a CDR3 comprising a sequence set forth in amino acids 89-97 of SEQ ID NO: 5.

[0166] In some examples, the antibody comprises a VH and a VL, wherein:

[0167] (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:

[0168] Xi is selected from the group consisting of tryptophan, glutamine, methionine, serine, phenylalanine, glutamic acid and histidine;

[0169] X2 is an amino acid selected from the group consisting of phenylalanine, tyrosine, methionine, serine, glycine and isoleucine;

[0170] X3 is an amino acid selected from the group consisting of aspartic acid, methionine, glutamine, serine, leucine, valine, arginine and histidine; and

[0171] 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

[0172] (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 ID NO: 13) wherein:

[0173] Xi is an amino acid selected from the group consisting of glutamine, glutamic acid, histidine, alanine and serine;

[0174] X2 is an amino acid selected from the group consisting of glutamine, valine, phenylalanine, asparagine and glutamic acid;

[0175] X3 is an amino acid selected from the group consisting of serine and glycine;

[0176] X4 is an amino acid selected from the group consisting of tryptophan, methionine, phenylalanine, tyrosine, isoleucine and leucine; Xs 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;

[0177] Xe is an amino acid selected from the group consisting of tyrosine, methionine, isoleucine and threonine;

[0178] X7 is an amino acid selected from the group consisting of proline, alanine, histidine, glycine and lysine;

[0179] 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.

[0180] In some examples, the antibody comprises a VH and a VL, wherein:

[0181] (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

[0182] (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.

[0183] In some examples, the antibody comprises a VH and a VL, wherein:

[0184] (A) (i) the VH comprises a CDR1 comprising a sequence set forth in amino acids 31-35 of

[0185] SEQ ID NO: 2; a CDR2 comprising a sequence set forth in amino acids 50-65 of SEQ ID NO: 2; and a CDR3 comprising a sequence set forth in amino acids 99-107 of SEQ ID NO: 2; and

[0186] (ii) the VL comprises a CDR1 comprising a sequence set forth in amino acids 24-34 of SEQ ID NO: 3; a CDR2 comprising a sequence set forth in amino acids 51-56 of SEQ ID NO: 3; and a CDR3 comprising a sequence set forth in amino acids 89-97 of SEQ ID NO: 3; or

[0187] (B) (i) the VH comprises a CDR1 comprising a sequence set forth in amino acids 31-35 of

[0188] SEQ ID NO: 4; a CDR2 comprising a sequence set forth in amino acids 50-65 of SEQ ID NO: 4; and a CDR3 comprising a sequence set forth in amino acids 99-107 of SEQ ID NO: 4; and

[0189] (ii) the VL comprises a CDR1 comprising a sequence set forth in amino acids 24-34 of SEQ ID NO: 5; a CDR2 comprising a sequence set forth in amino acids 51-56 of SEQ ID NO: 5; and a CDR3 comprising a sequence set forth in amino acids 89-97 of SEQ ID NO: 5; or

[0190] (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

[0191] (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.

[0192] In some examples, the antibody comprises:

[0193] (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;

[0194] (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;

[0195] (iii) a VH comprising three CDRs of a VH comprising an amino acid sequence set forth in

[0196] 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;

[0197] (iv) a VH comprising three CDRs of a VH comprising an amino acid sequence set forth in

[0198] 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

[0199] (v) a VH comprising:

[0200] (a) a CDR1 comprising a sequence set forth in SEQ ID NO: 6;

[0201] (b) a CDR2 comprising a sequence set forth in SEQ ID NO: 7; and

[0202] (c) a CDR3 comprising a sequence set forth in SEQ ID NO: 8; and a VL comprising:

[0203] (a) a CDR1 comprising a sequence set forth in SEQ ID NO: 9;

[0204] (b) a CDR2 comprising a sequence set forth in SEQ ID NO: 10; and

[0205] (c) a CDR3 comprising a sequence set forth in SEQ ID NO: 11.

[0206] In some examples, the antibody comprises a human constant region, e.g., an IgG constant region, such as an IgGi, IgG2, IgGa or IgG4 constant region or mixtures thereof. In the case of an antibody 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.

[0207] In some examples, the antibody comprises a constant region of an IgG4 antibody or a stabilized constant region of an IgG4 antibody. In some examples, the antibody comprises an IgG4constant 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)). The C-terminal lysine of the heavy chain constant region of a whole antibody (or an antibody comprising a constant region or a CH3) may be removed, for example, during production or purification of the antibody, 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.

[0208] In some examples the antibody or a composition comprising the antibody, 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.

[0209] In some examples, the antibody 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.

[0210] In some examples, the antibody comprises:

[0211] (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

[0212] (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.

[0213] In some examples, the antibody comprises 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 antibody comprises 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 antibody comprises 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.

[0214] In some examples, the antibody comprises 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.

[0215] In some examples, the antibody is any form of the antibody encoded by a nucleic acid encoding any of the foregoing antibodies. In this context, “encoded by” refers to an antibody amino acid sequence encoded by a nucleic acid sequence, before any post-translational modification (e.g., deamidation etc) to the amino acid sequence. In some examples, the antibody comprises a VH encoded by a nucleic acid encoding an amino acid of SEQ ID NO: 4 and a VL encoded by a nucleic acid encoding an amino acid of SEQ ID NO: 5.

[0216] In some examples, the antibody comprises a VH encoded by a nucleic acid encoding an amino acid of SEQ ID NO: 2 and a VL encoded by a nucleic acid encoding an amino acid of SEQ ID NO: 3.

[0217] In some examples, the antibody comprises:

[0218] (i) a VH comprising:

[0219] (a) a CDR1 encoded by a nucleic acid encoding an amino acid of SEQ ID NO: 6;

[0220] (b) a CDR2 encoded by a nucleic acid encoding an amino acid of SEQ ID NO: 7; and

[0221] (c) a CDR3 encoded by a nucleic acid encoding an amino acid of SEQ ID NO: 8; and

[0222] (ii) a VL comprising:

[0223] (a) a CDR1 encoded by a nucleic acid encoding an amino acid of SEQ ID NO: 9;

[0224] (b) a CDR2 encoded by a nucleic acid encoding an amino acid of SEQ ID NO: 10; and

[0225] (c) a CDR3 encoded by a nucleic acid encoding an amino acid of SEQ ID NO: 11.

[0226] In some examples, the antibody comprises:

[0227] (i) a VH comprising:

[0228] (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 23;

[0229] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 24; and

[0230] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 25; and

[0231] (ii) a VL comprising:

[0232] (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 26;

[0233] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 27; and

[0234] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 28.

[0235] In some examples, the antibody comprises:

[0236] (i) a VH comprising:

[0237] (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

[0238] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 25; and

[0239] (ii) a VL comprising:

[0240] (a) a CDR1 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 26;

[0241] (b) a CDR2 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 27; and

[0242] (c) a CDR3 comprising a sequence encoded by a nucleic acid comprising SEQ ID NO: 28.

[0243] In some examples, the antibody 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.

[0244] In some examples, the antibody 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.

[0245] In some examples, the antibody 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.

[0246] In some examples, the antibody is anumigilimab (also referred to herein as “CSL324” or “C1.2G”).

[0247] An exemplary method disclosed herein is a method of treating moderate to severe hidradenitis suppurativa (HS) in a subject suffering from HS, wherein the subject is non- responsive to treatment with adalimumab, the method comprising administering subcutaneously to the subject a fixed dose of 25 mg or 50 mg of anumigilimab every 13-15 days. As described herein, the antibody may be administered every 14 days.

[0248] In some examples, the antibody is administered in a pharmaceutical formulation. In some examples, the pharmaceutical formulation comprises the antibody at a concentration of 50 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and comprises 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 mM proline, and 90 mM to 110 mM arginine.

[0249] In some examples, the pharmaceutical formulation comprises the antibody at a concentration of 50 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and comprises about 20 mM histidine buffer, about 0.03% (w / v) polysorbate 80, about 100 mM proline, and about 100 mM arginine. In some examples, the pharmaceutical formulation comprises the antibody at a concentration of 50 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and comprises 20 mM histidine buffer, 0.03% (w / v) polysorbate 80, 100 mM proline, and 100 mM arginine.

[0250] In some examples, the pharmaceutical formulation consists of:

[0251] (i) the antibody at a concentration of 50 mg / mL;

[0252] (ii) 20 mM L-histidine buffer;

[0253] (iii) 0.03% (w / v) polysorbate 80;

[0254] (iv) 100 mM L-arginine-HCl; and

[0255] (v) 100 mM L-proline, wherein the formulation has a pH of 5.7.

[0256] In some examples, the human subject is an adult, for example 18 years of age or older. In some examples, the human subject is a child, for example less than 18 years of age.

[0257] An exemplary method disclosed herein is a method of treating HS in a human subject suffering from the HS, the method comprising administering subcutaneously to the subject a fixed dose of 25 mg or 50 mg of an antibody that binds to G-CSFR every 13-15 days, wherein the antibody comprises 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, and wherein the antibody is administered in a pharmaceutical formulation comprising the antibody at a concentration of 50 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and comprises 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 mM proline, and 90 mM to 110 mM arginine.

[0258] An exemplary method disclosed herein is a method of treating moderate to severe HS in a human subject suffering from the HS, wherein the subject has had an inadequate response to an anti-tumour necrosis factor (TNF) therapy, the method comprising administering subcutaneously to the subject a fixed dose of 25 mg or 50 mg of anumigilimab every 13-15 days, wherein anumigilimab is administered in a pharmaceutical formulation comprising anumigilimab at a concentration of 50 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and comprises 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 mM proline, and 90 mM to 110 mM arginine.

[0259] In some examples a dose of 50 mg of anumigilimab is administered to the subject every 13-15 days. In some examples a dose of 25 mg of anumigilimab is administered to the subject every 13-15 days.

[0260] The present disclosure also provides a kit comprising an antibody that inhibits G-CSF signaling packaged with instructions for use in a method described herein.

[0261] In some examples, the kit optionally comprises an additional therapy for administration in use in combination with an antibody described herein. The present disclosure also provides an article of manufacture comprising a single dose container containing a single fixed dose of an antibody described herein formulated for subcutaneous administration, wherein the fixed dose is 50 mg or 25 mg. The article of manufacture may be packaged with instructions for use in a method described herein.

[0262] The features described in any of the above examples of the disclosure apply mutatis mutandis to all methods, uses, compositions for use, kits and articles of manufacture provided by the present disclosure. The above examples of the disclosure are not to be considered as excluding features described in other examples of the disclosure. Thus, features in different examples of the disclosure may be combined, unless the context requires otherwise.

[0263] BRIEF DESCRIPTION OF THE DRAWINGS

[0264] 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.

[0265] 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.

[0266] Figure 3 is an assessment of the enrichment score of Th 17 cells gene sets (panel A) and IL- 17 gene sets (panel B and C) 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.

[0267] 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.

[0268] 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. 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 nonsignificant.

[0269] Figure 6 is the overall study schema for Example 5. 1° EP = endpoint; Q2W = every 2 weeks.

[0270] Figure 7 is a schematic of the ANC model described in Example 4. IV = intravenous; SC = subcutaneous; VC = volume of the central compartment; VP = volume of the peripheral compartment; CL = linear clearance; Q = intercompartmental clearance; Vmax = maximum nonlinear elimination rate; Km = concentration achieving half of the maximum elimination rate (Michaelis-Menten constant); Kprol = rate constant for proliferating cells; Circ = circulating cells; CircO = baseline circulating cells; Ktr = transit rate constant; TRn = transit compartment; Kcirc = turnover rate constant for circulating cells; y = exponent of feedback loop.

[0271] Figure 8 shows a prediction-corrected visual predictive check for the ANC Model developed by the inventors (as described in Example 4). The model’s performance was tested against observed data in the CSL324_1002 trial in patients with HS or PPP (i.e., the trial described in Example 1) as well as healthy subjects. ANC = absolute neutrophil count; CI = confidence interval.

[0272] Figure 9 is a series of graphs showing simulated CSL324 concentration-time profiles and ANC-time profiles for subjects receiving 25 mg or 50 mg CSL324 Q2W SC dosing regimens. Q2W = every two weeks; IQR = inter-quartile range; PI = prediction interval; ANC = absolute neutrophil count. Solid black line is the median, dashed lines show the IQR, and shaded area represents the 90% PI. Horizontal dotted line on ANC profiles represents ANC = 1.5 x 109cells / L.

[0273] KEY TO SEQUENCE LISTING

[0274] DESCRIPTION

[0275] General

[0276] 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.

[0277] 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 antibodies 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.

[0278] 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.

[0279] 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).

[0280] 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.

[0281] 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). 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).

[0282] 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.

[0283] 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.

[0284] Any discussion of an antibody herein will be understood to include any variants of the 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.

[0285] 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.

[0286] 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.

[0287] 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.

[0288] 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.

[0289] 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.

[0290] Selected Definitions

[0291] 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.

[0292] 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.

[0293] 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 Laboratory 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: 2458-24642004) without affecting ligand binding.

[0294] 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 antibody, 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.

[0295] As used herein, the phrase “reduce” or “reducing” shall be understood to include administering an antibody described herein to make smaller or less in amount, degree or size at least one specified disease or condition, or symptom thereof.

[0296] As used herein, the phrase “inhibit” or “inhibiting” shall be understood to include administering an antibody described herein to eliminate or stop, at least partially the development of at least one specified disease or condition, or symptom thereof.

[0297] As used herein, the phrase “hinder” or “hindering” shall be understood to include administrating an antibody described herein to limit or impede or hamper, at least partially the development of at least one specified disease or condition, or symptom thereof.

[0298] 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 nonhuman primates. For example, the subject may be a human.

[0299] The term “reduce”, “prevent an increase in” or “inhibit” are used herein to refer to a lower amount of any of the genes and / or factors recited herein, relative to either the amount in the subject prior to administration of the antibody that inhibits G-CSF signaling, or relative to the amount in a corresponding control subject. For instance, the control subject may be a subject who receives a placebo and / or a standard of care therapy, rather than the antibody that inhibits G-CSF signaling.

[0300] The term “increase”, “prevent a decrease in” are used herein to refer to a higher amount of any of the genes and / or factors recited herein, relative to either the amount in the subject prior to administration of the antibody that inhibits G-CSF signaling, or relative to the amount in a corresponding control subject. For instance, the control subject may be a subject who receives a placebo and / or a standard of care therapy, rather than the antibody that inhibits G- CSF signaling.

[0301] 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:

[0302] • Grade 1: < 2.0 x 109 / L (< 2000 / mm3) and > 1.1 x 109 / L (> 1500 / mm3)

[0303] • Grade 2: < 1.5 x 109 / L (< 1500 / mm3) and > 1.0 x 109 / L (> 1000 / mm3)

[0304] • Grade 3: < 1.0 x 109 / L (< 1000 / mm3) and > 0.5 x 109 / L (> 500 / mm3)

[0305] • Grade 4: < 0.5 x 109 / L (< 500 / mm3).

[0306] 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.

[0307] 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.

[0308] 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.

[0309] 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.

[0310] 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.

[0311] 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.

[0312] 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 A light chain and a heavy chain from mammals is a, 5, £, y, or p. Antibodies can be of any type (e.g., IgG, IgE, IgM, IgD, IgA, and IgY), class (e.g., IgGi, IgG2, IgGs, IgG4, IgAi and IgA2) or subclass. The term “antibody” also encompasses humanized antibodies, primatized antibodies, human antibodies and chimeric antibodies.

[0313] 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 wild-type sequence constant domains (e.g., human wild-type sequence constant domains) or amino acid sequence variants thereof.

[0314] 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.

[0315] 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 some examples, reference herein to a CDR (or a FR) is in respect of those regions according to the Kabat numbering system.

[0316] "Framework regions" (FRs) are those variable region residues other than the CDR residues.

[0317] 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, a F(ab’) fragment, a scFv, a diabody, a triabody, 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 “Fab2” 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.

[0318] As used herein, the term “binds” in reference to the interaction of an antibody 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.

[0319] As used herein, the term “specifically binds” or “binds specifically” shall be taken to mean that an antibody described herein 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, an antibody 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.

[0320] An 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 some examples, an 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.

[0321] 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.

[0322] 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.

[0323] 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.

[0324] 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.

[0325] 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 some examples, the competitive inhibition is not due to steric hindrance.

[0326] “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.

[0327] As used herein, the term “neutralize” shall be taken to mean that an antibody 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.

[0328] As used herein, the term “about”, in relation to a numerical value provided for a given parameter, refers to values within + / - 10%, or preferably within + / - 5% or + / -2%, of the recited value. Use of the term “about” is also intended to provide explicit basis for the precise value indicated. For example, the phrase “about 50 mg” is intended to encompass “about 50 mg” or “50 mg”.

[0329] Neutrophilic skin conditions

[0330] The present disclosure relates to methods of treating neutrophilic skin conditions. As used herein, the term “neutrophilic 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.

[0331] Exemplary neutrophilic skin conditions include neutrophilic dermatoses such as hidradenitis suppurativa (HS); palmoplantar pustulosis (PPP); amicrobial pustulosis of the folds (APF); psoriasis; plaque psoriasis; CARD14-mediated pustular psoriasis (CAMPS); cryopyrin associated periodic syndromes (CAPS); deficiency of interleukin- 1 receptor (DIRA); deficiency of interleukin-36 receptor antagonist (DIRTA); palmoplantar pustulosis (PPP); pyogenic arthritis; pyoderma gangrenosum and acne (PAPA); pyoderma gangrenosum, acne, and hidradenitis suppurativa (PASH); pyoderma gangrenosum(PG); skin lesions of Behcet’s disease; Still’s disease; Sweet syndrome; subcorneal pustulosis (Sneddon-Wilkinson); pustular psoriasis; acute generalized exanthematic pustulosis; infantile acropustulosis; synovitis, acne, pustulosis; hyperostosis and osteitis (SAPHO) syndrome; bowel-associated dermatosisarthritis syndrome (BAD AS); neutrophilic dermatosis of the dorsal hands; neutrophilic eccrine hidradenitis; erythema elevatum diutinum; and Pyoderma gangrenosum.

[0332] In some examples, the neutrophilic 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.

[0333] Certain subtypes of HS may be treated in accordance with the present disclosure. In one instance, moderate to severe HS, is treated by administering an antibody disclosed herein. In some examples, chronic HS, e.g., moderate to severe chronic HS, is treated by administering an antibody disclosed herein. In some examples, the subject has confirmed moderate to severe HS, e.g., confirmed chronic moderate to severe HS.

[0334] The present disclosure also provides a method for treating certain subpopulations of HS patients who may be especially difficult to treat. For example, in one instance, the present disclosure provides a method for treating patients who have a subtherapeutic response to a therapy, such as those who have been unresponsive or intolerant to oral antibiotics for treatment for their HS.

[0335] The present disclosure also provides methods for improving HS symptoms in a subject based on indices used to measure the disease state.

[0336] Treatment of HS using an antibody disclosed herein may also be determined using measures known in the art. Treatment of HS 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.

[0337] 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.

[0338] In one instance, treatment of HS with an antibody 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 TNFa inhibitor. In one instance, improvement in a Hurley score indicates that the Hurley score of the subject has either improved or been maintained following treatment with an antibody.

[0339] 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.

[0340] 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, 171 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.

[0341] 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.

[0342] In some examples, treatment of HS with an antibody disclosed herein is determined according to an 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.

[0343] In another example, treatment of HS with an antibody disclosed herein is defined as achieving an Physician's Global Assessment (PGA) score as defined in Table 1 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.

[0344] Table 1. PGA Scoring.

[0345] In one instance, 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.

[0346] In some examples, 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.

[0347] In some examples, the neutrophilic 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.

[0348] 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.

[0349] The present disclosure also provides a method for treating certain subpopulations of PPP patients who may be especially difficult to treat. For example, in one instance, the present disclosure provides a method for treating patients who have a subtherapeutic response to a therapy, such as those who have been unresponsive or topical corticosteroids, vitamin D3 analogues, etretinate, and phototherapy for treatment for their PPP.

[0350] The present disclosure also provides methods for improving PPP symptoms in a subject based on indices used to measure the disease state. Treatment of PPP using an antibody disclosed herein may be determined using measures known in the art. Treatment of PPP may be determined using any of the measures known in the art, e.g., improvement in ppPASI, or any measure known to those in the art.

[0351] 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 an antibody 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.

[0352] In one example, administration of an antibody 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.

[0353] 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).

[0354] Antibodies

[0355] The methods provided herein comprise administering an antibody that inhibits G-CSF signaling. In some examples, the antibody comprises a VH and a VL. In some examples, the antibody binds to G-CSFR. Exemplary antibodies that bind to G-CSFR and inhibit G-CSF signaling are described in WO2012 / 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.

[0356] 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.

[0357] 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.

[0358] 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. 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).

[0359] 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.

[0360] 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 de -immunized antibody.

[0361] In some examples, 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.

[0362] The antibodies of the present disclosure may be humanized or human.

[0363] 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 antigen -binding 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.

[0364] 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.

[0365] 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.

[0366] 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 are described in W02012171057, which is incorporated herein by reference. One particular exemplary antibody is anumigilimab (also referred to herein as “CSL324”).

[0367] Antibody Binding Domain Containing Proteins

[0368] Single-Domain Antibodies

[0369] In some examples, an antibody described herein is a protein that is or comprises a singledomain 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).

[0370] Diabodies, Triabodies, Tetrabodies

[0371] In some examples, the antibody is or comprises a diabody, triabody, tetrabody or higher order protein complex such as those described in WO98 / 044001 and / or W094 / 007921.

[0372] Single Chain Fv (scFv)

[0373] In some examples, the antibody is or comprises a scFv. 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.

[0374] Heavy Chain Antibodies

[0375] 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).

[0376] 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.

[0377] 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. Other Antibodies and Antibody Fragments

[0378] The present disclosure also contemplates other antibodies and antibody fragments, such as:

[0379] (i) “key and hole” bispecific proteins as described in US5,731,168;

[0380] (ii) heteroconjugate proteins, e.g., as described in US4,676,980;

[0381] (iii) heteroconjugate proteins produced using a chemical cross-linker, e.g., as described in US4,676,980; and

[0382] (iv) Faba (e.g., as described in EP19930302894).

[0383] De-immunized Antibodies and Proteins

[0384] The present disclosure also contemplates a de-immunized antibody. De-immunized antibodies 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 W02000 / 34317, W02004 / 108158 and W02004 / 064724.

[0385] Methods for introducing suitable mutations and expressing and assaying the resulting protein will be apparent to the skilled artisan based on the description herein.

[0386] Mutations to Proteins

[0387] The present disclosure also contemplates mutant forms of an antibody described herein. For example, amino acids at sites within a CDR of an antibody disclosed herein may be substituted. The skilled person will understand that changes can additionally or alternatively be made within a framework region of a variable region without inhibiting or significantly reducing its function in the context of the present disclosure.

[0388] For example, such a mutant antibody may comprise one or more conservative amino acid substitutions compared to a sequence set forth herein. In some examples, the antibody 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.

[0389] In some examples, a mutant antibody has only, or not more than, one or two or three or four or five or six conservative amino acid changes within its variable regions when compared to antibody disclosed herein. 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 antibody. 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).

[0390] The present disclosure also contemplates non-conservative amino acid changes (e.g., substitutions) in an antibody 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 some examples, 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 CDR, such as in a CDR3.

[0391] 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.

[0392] Constant Regions

[0393] The antibody used in accordance with the methods of the disclosure may comprise a constant region of an antibody. This includes antigen binding fragments of an antibody fused to a Fc. Thus, in some examples, the antibody comprises an Fc.

[0394] 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 some examples, the constant region is human isotype IgG4 or a stabilized IgG4 constant region.

[0395] In some examples, 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 some examples, 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.

[0396] In some examples, 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). In some examples, 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.

[0397] In some examples, 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).

[0398] 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).

[0399] 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, L234V, L235A and deletion of G236 and / or one or more of the following changes A327G, A330S and P331S (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).

[0400] 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 WO2010 / 085682). Exemplary substitutions include 240F, 262L, 264T, 266F, 297Q, 299A, 299K, 307P, 309K, 309M, 309P, 323F, 399S, and 427F.

[0401] Additional Modifications / Variants

[0402] The present disclosure also contemplates additional modifications or variants to an antibody disclosed herein.

[0403] For example, the antibody may comprise one or more amino acid substitutions that increase the half-life of the protein. For example, the antibody may comprise 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 may have increased affinity for FcRn at lower pH, e.g., about pH 6.0, to facilitate Fc / FcRn binding in an endosome. In some examples, 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.

[0404] 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.

[0405] In some examples, an antibody described herein additionally comprises albumin, a functional fragment or variant thereof. In some examples, the albumin, functional fragment or variant thereof is serum albumin, such as human serum albumin. In some examples, 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, WO2011103076, WO20 12112188, WO2013075066, WO2015063611 and WO2014179657.

[0406] In some examples, the antibody described herein comprises one or more variants. For example, the variant is a post-translationally modified variant.

[0407] In some examples, the 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.

[0408] In some examples, the antibody comprises a variant missing an encoded C-terminal lysine residue.

[0409] In some examples, the 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.

[0410] In some examples, the antibody comprises a misincorporated amino acid residue. For example, methionine residues may be substituted with nor-leucine, asparagine residues may be substituted with serine and / or phenylalanine residues may be substituted with tyrosine.

[0411] In some examples, the antibody comprises a variant comprising a pyroglutamate. For example, at the N-terminus of a protein.

[0412] In some examples, the antibody comprises a glycosylated variant.

[0413] In some examples, the antibody comprises a variant lacking a N-terminal residue. For example, a N-terminal glutamine in an antibody or V region.

[0414] In some examples, the 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).

[0415] Protein Production

[0416] Antibodies described herein may be produced by any method known in the art. For example, the antibody may be produced by culturing a hybridoma under conditions sufficient to produce the protein, e.g., as described herein and / or may be produced via recombinant expression. Recombinant Expression

[0417] An antibody described herein according to any example is recombinant. 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.

[0418] 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.

[0419] 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.

[0420] 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.

[0421] 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).

[0422] Exemplary promoters active in mammalian cells include cytomegalovirus immediate early promoter (CMV-IE), human elongation factor 1-oc promoter (EFl), small nuclear RNA promoters (Ula and Ulb), oc-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).

[0423] 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 GAL4 promoter, the CUP1 promoter, the PHO 5 promoter, the nnit promoter, the RPR1 promoter, or the TEF1 promoter.

[0424] 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.

[0425] 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.

[0426] Isolation of Proteins

[0427] Methods for isolating a protein are known in the art and / or described herein.

[0428] 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.

[0429] 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).

[0430] 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 a 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.

[0431] Assaying Activity of an antibody

[0432] Binding to G-CSFR and Mutants Thereof

[0433] It will be apparent to the skilled artisan from the disclosure herein that some antibodies 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 may involve labeling the protein and contacting it with immobilized antibody. 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 antibody 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).

[0434] 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 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.

[0435] 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.

[0436] In some examples, proteins having a similar Kd or a higher Kd than Cl.2 or C1.2G are selected, because they are likely to compete for binding to hG-CSFR.

[0437] The assays described above can also be used to detect the level of binding of an antibody to hG-CSFR or a ligand binding domain thereof (e.g., SEQ ID NO: 1) or mutant form thereof.

[0438] The level of binding may be conveniently determined using a biosensor, for example.

[0439] In some examples, an antibody described herein 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.

[0440] In some examples, an antibody described herein 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 some examples, an antibody described herein 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.

[0441] In some examples, an antibody described herein 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 some examples, an antibody described herein 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.

[0442] In some examples, an antibody described herein 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 some examples, an antibody described herein 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.

[0443] In some examples, an antibody described herein 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 some examples, an antibody described herein 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.

[0444] In some examples, an antibody described herein 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 some examples, an antibody described herein 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.

[0445] In some examples, an antibody described herein 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 some examples, an antibody described herein 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.

[0446] In some examples, an antibody described herein 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.

[0447] In some examples, an antibody described herein 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.

[0448] The present disclosure contemplates any combination of the foregoing characteristics. In some examples, an antibody described herein has all of the binding characteristics set forth in the preceding seven paragraphs.

[0449] Epitope Mapping

[0450] In another example, the epitope bound by an antibody 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 noncontiguous peptides are bound by the protein, the protein may bind a conformational epitope. 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.

[0451] A further method is exemplified herein and involves binding hG-CSFR or a region thereof to an immobilized antibody described herein 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.

[0452] A further method involves converting hydrogens in hG-CSFR or a region thereof to deutrons and binding the resulting protein to an immobilized antibody described herein. 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 an antibody described herein.

[0453] Alternatively or additionally, the epitope of an antibody described herein may be determined using x-ray crystallography using methods well known in the art.

[0454] Determining Competitive Binding

[0455] 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.

[0456] 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.

[0457] 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 Cl.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 C 1.2 or C 1.2G 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 C 1.2 or C 1.2G to hG-CSFR.

[0458] 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 C1.2 or C1.2G binds, e.g., as described herein.

[0459] Determining Neutralization

[0460] In some examples of the present disclosure, an antibody is capable of neutralizing hG- CSFR signaling. In this regard, the term “neutralizing” is used synonymously with “inhibiting” and does not require 100% inhibition of G-CSFR signalling.

[0461] Various assays are known in the art for assessing the ability of an antibody to neutralize signaling of a ligand through a receptor.

[0462] In some examples, the antibody 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.

[0463] In another example, the antibody 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 antibody. 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 antibody that inhibits G-CSF signaling compared to in the absence of the antibody that inhibits G-CSF signaling indicates that the antibody that inhibits G-CSF signaling neutralizes G-CSF signaling. By testing multiple concentrations of the antibody 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 some examples, 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 some examples, 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.

[0464] In a further example, the antibody 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 antibody. Methods for assessing cell proliferation are known in the art and include, for example, MTT reduction and thymidine incorporation. An antibody that reduces the level of proliferation compared to the level observed in the absence of the antibody is considered to neutralize G- CSF signaling. By testing multiple concentrations of the antibody an ICso is determined, i.e., a concentration at which 50% of the maximum inhibition of cell proliferation occurs. In some examples, the ICso is 6nM or less, such as 5.9nM or less. In another example, the ICso 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.

[0465] In a further example, the antibody 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 antibody 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. An antibody that reduces the level of the cell(s) compared to the level observed in the absence of the antibody is considered to neutralize G-CSF signaling. In some examples, the antibody that inhibits G-CSF signaling reduces the number of neutrophils without inducing neutropenia.

[0466] Other methods for assessing neutralization of G-CSF signaling are contemplated by the present disclosure.

[0467] Determining Effector Function

[0468] As discussed herein, some proteins of the present disclosure have reduced effector function. Methods for assessing ADCC activity are known in the art.

[0469] In some examples, 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 antibodies that inhibit G-CSF signaling for a time and under conditions sufficient for the antibody 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 antibody 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 83:1059-1063, 1986 and Bruggemann, et al., J. Exp. Med. 166: 1351-1361, 1987.

[0470] 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).

[0471] 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.

[0472] Determining Half Life

[0473] 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. 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 (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, 1994).

[0474] 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 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.

[0475] Efficacy

[0476] The efficacy of an antibody that inhibits G-CSF signaling to treat a neutrophilic skin condition can be assessed using any method known in the art. For example, the effectiveness of the treatment may be assessed by comparing the degree of severity of the disease or symptoms in subjects administered with the antibody relative to subjects not administered the antibody. Alternatively, or additionally, therapeutic efficacy of candidate antibodies can be assessed in an animal model.

[0477] 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). 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. Efficacy can also be assessed by quantifying the subject’s 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.

[0478] Efficacy can be assessed by determining a IHS4 score for the subject before and after treatment with the antibody. As described herein, the IHS4 score 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. In some examples, treatment in accordance with a method of the disclosure results in a reduction in the subject’s IHS4 score.

[0479] Efficacy of treatment can be assessed by determining whether the subject achieves the criteria of IHS4-55. IHS4-55 is a recently reported dichotomous outcome measure that is defined as a > 55% decrease from baseline in IHS4 score.

[0480] Efficacy of treatment can be assessed by determining whether the subject achieves a Hidradenitis Suppurative Clinical Response (HiSCR), also referred to herein as “HiSCR50”, as described in (Kimball et al, 2014, Br J Dermatol. 171 : 1434). The HiSCR (HiSCR50) is defined as a > 50% reduction from baseline in the total abscess and nodule count with no increase in abscess or draining tunnel count. 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. In some examples, treatment in accordance with a method of the disclosure results in the subject achieving a HiSCR50. In some examples, treatment in accordance with a method of the disclosure results in the subject achieving a HiSCR75.

[0481] Modified HiSCR is another response criteria recently reported [InflaRX, 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.

[0482] Efficacy of treatment can 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).

[0483] Efficacy of treatment can be assessed using DLQI, which is a validated questionnaire consisting of ten questions that assess the effect of skin conditions on the quality of life of an affected patient (Finlay et al. Clin Exp Dermatol. 1994, 19:210). Each question is scored from 0 (not at all) to 3 (very much), with a recall period of 1 week, and scores are tallied to produce a total score. Total scores are graded as follows: 0 to 1 indicates no effect, 2 to 5 indicates a small effect, 6 to 10 indicates a moderate effect, 11 to 20 indicates a very large effect, and 21 to 30 (maximum score) indicates an extremely large effect on the patient’s life. In some examples, treatment in accordance with a method of the disclosure results in the subject achieving a reduction in DLQI.

[0484] Efficacy of treatment may also be assessed by determining the level of skin pain experienced by a subject. Determining skin pain may be performed using a pain numerical rating scale (NRS). The pain NRS is a unidimensional measure of pain intensity in adults, including those with chronic pain associated with skin conditions (Kimball et al. 2016, 375:422). The reduction in the patient’s global assessment of skin pain (e.g., assessed using the pain NRS) may be: a) a reduction in pain at its worst and pain on average from baseline; and / or b) a reduction pain at its worst and pain on average over the previous 24 hours; and / or b) a reduction in pain at its worst and pain on average over the previous 7 days.

[0485] Efficacy of treatment may also be assessed by determining the subject’s Hi dradenitis Suppurativa Quality of Life score (HiSQOL), as described in Kirby et al., Br J Dermatol, 2020, 183:340. The 17-item HiSQOL included 4 symptom items, 8 activity adaptation items, and 5 psychosocial items. Each question is scored from 4 (extremely) to 0 (not at all), with a recall period of 1 week. The item scores are summed to create a total score ranging from 0 to 68, with higher scores indicating more severe impact on health-related quality of life. The subscale scores range from 0 to 16 for symptoms, 0 to 20 for psychosocial, and 0 to 32 for activities- adaptations. In some examples, treatment in accordance with a method of the disclosure results in a reduction in the subject’s HiSQOL score.

[0486] In some examples, assessing the efficacy of an antibody 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.

[0487] 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.

[0488] 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.

[0489] Compositions and administration

[0490] The methods of the disclosure comprise administering an antibody subcutaneously to a subject. The antibody may be administered in a pharmaceutical formulation. Methods for preparing an antibody 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).

[0491] The compositions for administration will commonly comprise a solution of the antibody 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 antibody 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. Upon formulation, antibodies of the present disclosure may be administered in a manner compatible with the dosage formulation and in such amount as is prophylactically or therapeutically effective.

[0492] A formulation comprising the antibody may comprise one or more or all of a histidine buffer, a polysorbate (e.g., polysorbate 80), proline and arginine. The pH may be in the range of 5 to 6, for example, or in the range of 5.5 to 5.9.

[0493] In some examples, the formulation comprises a histidine buffer. Suitable histidine buffers for use in the present disclosure will be apparent to the skilled person and include, for example L-histidine, histidine chloride, histidine acetate, histidine phosphate, histidine sulfate, etc. In one example, the histidine buffer is L-histidine.

[0494] In some examples, the formulation comprises 12 mM to 30 mM histidine buffer. For example, the formulation may comprise 15 mM to 25 mM histidine buffer. For example, the formulation may comprise 17 mM to 23 mM histidine buffer. For example, the formulation may comprise 18 mM to 22 mM histidine buffer. For example, the formulation may comprise 15 mM, 16 mM, 17 mM, 18 mM, 19 mM, 20 mM, 21 mM, 22 mM, 23 mM, 24 mM, or 25 mM histidine buffer. For example, the formulation may comprise 20 mM histidine buffer.

[0495] Methods of assessing the suitability of buffers will be apparent to the skilled person and / or described herein and include, for example, differential scanning fluorimetry and dynamic light scattering.

[0496] In some examples, the formulation comprises polysorbate 80. Polysorbate 80 (polyoxyethylene sorbitan monooleate) is a non-ionic surfactant that can be added to the pharmaceutical formulations in an amount such that it suppresses aggregation (e.g., by preventing surface denaturation), increases stabilisation (e.g., during thermal and / or physical stress), minimises the formation of particulates in the formulation (e.g., sub-visible and / or visible particle formation), reduces surface adsorption and / or assists in protein refolding.

[0497] In some examples, the formulation comprises 0.01 % to 0.05 % (w / v) polysorbate 80. For example, the formulation may comprise of 0.02% (w / v) to 0.04 %(w / v) polysorbate 80. For example, the formulation may comprise 0.01% (w / v), 0.02% (w / v), 0.03% (w / v), 0.04% (w / v), or 0.05% (w / v) polysorbate 80. For example, the formulation may comprise 0.03% (w / v) polysorbate 80.

[0498] In some examples, the formulation comprises one or more amino acid stabilisers. For example, the formulation may comprise proline and / or arginine. The amino acid stabilisers may be added to the pharmaceutical formulation in an amount such that they reduce thermal and / or physical stress (e.g., freeze / thaw or agitation), and / or confer or enhance stability of the antibody.

[0499] In some examples, the formulation comprises proline and arginine or a salt form thereof. In some examples, the formulation comprises 95 mM to 110 mM proline. For example, the formulation may comprise 100 mM to 110 mM proline. For example, the formulation may comprise 100 mM, 101 mM, 102 mM, 103 mM, 104 mM, 105 mM, 106 mM, 107 mM, 108 mM, 109 mM, or 110 mM proline. For example, the formulation may comprise 100 mM proline.

[0500] In some examples, the formulation comprises 60 mM to 125 mM arginine. For example, the formulation 90 mM to 110 mM arginine. For example, the formulation may comprise 95 mM to 105 mM arginine. For example, the formulation may comprise 95 mM, 96 mM, 97 mM, 98 mM, 99 mM, 100 mM, 101 mM, 102 mM, 103 mM, 104 mM, or 105 mM arginine. For example, the formulation may comprise 100 mM arginine.

[0501] In some examples, the formulation comprises 100 mM to 125 mM proline and 60 mM to 125 mM arginine. For example, the formulation may comprise 100 mM to 110 mM proline and 90 mM to 110 mM arginine. For example, the formulation may comprise 100 mM, 101 mM, 102 mM, 103 mM, 104 mM, 105 mM, 106 mM, 107 mM, 108 mM, 109 mM, or 110 mM proline; and 95 mM, 96 mM, 97 mM, 98 mM, 99 mM, 100 mM, 101 mM, 102 mM, 103 mM, 104 mM, or 105 mM arginine. For example, the formulation may comprise 100 mM proline and 100 mM arginine.

[0502] Pharmaceutical formulations described herein may comprise the antibody at any suitable concentration, for example at a concentration suitable for subcutaneous injection. In some examples, the antibody is present at a concentration of 45 mg / mL to 55 mg / mL. For example, the formulation may comprise the antibody at a concentration of 45 mg / mL, 46 mg / mL, 47 mg / mL, 48 mg / mL, 49 mg / mL, 50 mg / mL, 51 mg / mL, 52 mg / mL, 53 mg / mL, 54 mg / mL, or 55 mg / mL. In some examples, the formulation comprises the antibody at a concentration of 50 mg / mL.

[0503] In some examples, the pH of the formulation is in the range of 5.5 to 5.9. For example, the pH may be 5.5, 5.6, 5.7, 5.8 or 5.9. In some examples, the pH is about 5.7.

[0504] An exemplary formulation comprises the antibody at a concentration of 50 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and comprises 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 mM proline, and 90 mM to 110 mM arginine. In some examples, the formulation comprises:

[0505] (i) 20 mM histidine buffer;

[0506] (ii) 0.03% (w / v) polysorbate 80;

[0507] (iii) 100 mM arginine; and / or

[0508] (iv) 100 mM proline.

[0509] An exemplary formulation therefore comprises or consists of:

[0510] (i) an antibody described herein (e.g., anumigilimab) at a concentration of 50 mg / mL;

[0511] (ii) 20 mM histidine buffer (e.g., L-histidine); (iii) 0.03% (w / v) polysorbate 80;

[0512] (iv) 100 mM arginine (e.g., L-arginine-HCl); and

[0513] (v) 100 mM proline (e.g., L-proline), wherein the formulation has a pH in the range of 5.5 to 5.9 (e.g., about 5.7).

[0514] The methods of the disclosure comprise administering a fixed dose of an antibody that inhibits G-CSF signaling. A "fixed dose" herein refers to a dosage of antibody which is administered without regard to the patient's weight or body surface area (BSA), i.e. it is not administered as either a mg / kg or mg / m2dose.

[0515] The doses of the antibody, in accordance with the methods of the invention, are administered subcutaneously. As used herein, the term “subcutaneous”, in the context of administration of an antibody to a subject, is used to refer to a method of inserting a composition comprising the antibody beneath the skin of the subject, for example by injection or infusion. In some examples, the antibody is administered by subcutaneous injection. As is known in the art, subcutaneous injection is an injection delivered as a bolus into the subcutis, the layer of skin directly below the dermis and epidermis, collectively referred to as the cutis. The injection may be delivered using any suitable means, for example using a hypodermic needle and a syringe, an injection device, an injector pen, a needleless device, an autoinjector or a subcutaneous patch delivery system. The doses of the antibody may thus be self-administered in some examples of methods of the disclosure.

[0516] The methods of the invention comprise administering the antibody subcutaneously at a fixed dose of 25 mg or 50 mg every 13-15 days. The “dose” may comprise one or more injections on the same day. For example, a dose of 50 mg may be administered as a single injection of 50 mg or two injections of 25 mg or any other suitable combination as determined by a medical practitioner. In some examples, each dose is administered as a single subcutaneous injection.

[0517] In some examples, the antibody or antibody is administered for a set period or number of doses. For example, the antibody or antibody is administered for 1 month or 3 months or 6 months or 12 months. In another example, five or 10 or 15 or 20 doses of the antibody or antibody is administered. For example, the antibody may be administered for a period of at least 16 weeks. In some examples, the antibody is administered for a period of at least 32 weeks.

[0518] In another example, the antibody or antibody is administered chronically or an on ongoing basis, e.g., for months or years and the present disclosure is not limited to a specific time period unless stated otherwise.

[0519] In another example, for subjects experiencing an adverse reaction, a dose may be split over numerous days in one week or over numerous consecutive days. In some examples, for subjects experiencing neutropenia as an adverse reaction, a caregiver may be instructed to cease treatment. For example, if the subject experiences neutropenia for more than 2 consecutive days or 3 consecutive days, treatment may be ceased.

[0520] In some examples, for subjects experiencing neutropenia as an adverse reaction, a caregiver may be instructed to skip the next dose. For example, if the subject experiences neutropenia for more than 2 consecutive days or 3 consecutive days, the next dose may be skipped.

[0521] Administration of an antibody according to the methods of the present disclosure can be continuous or intermittent, depending, for example, on the subject’s physiological condition, and other factors known to skilled practitioners. For example, the antibody described herein may be administered prior to, during or after development of systemic inflammation as described herein, or pro re nata, In some examples, the antibody is administered pro re nata.

[0522] In some examples, for subjects experiencing neutropenia as an adverse reaction, a caregiver may be instructed to cease treatment. For example, if the subject experiences neutropenia for more than 2 consecutive days or 3 consecutive days, treatment may be ceased.

[0523] In some examples, for subjects experiencing neutropenia as an adverse reaction, a caregiver may be instructed to skip the next dose. For example, if the subject experiences neutropenia for more than 2 consecutive days or 3 consecutive days, the next dose may be skipped.

[0524] Optionally, the subject suffering from neutropenia may be treated with G-CSF or GM- CSF to treat the neutropenia.

[0525] Combination Therapies

[0526] In some examples, the antibody 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.

[0527] For example, the other therapy may comporise 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.

[0528] In some examples, the antibody is administered simultaneously with the other compound. In some examples, the antibody that inhibits G-CSF signaling is administered before the other compound. In one example, the antibody that inhibits G-CSF signaling is administered after the other compound. In some examples, the antibody 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 antibody is administered in combination with a gene therapy.

[0529] In some examples, the antibody 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 antibody or compound can be administered over this time period. In one example, the antibody or compound is administered before the non-pharmaceutical intervention. In one example, the antibody is administered after the non-pharmaceutical intervention.

[0530] Another non-pharmaceutical intervention is light therapy. As is known in the art, light therapy is used to treat some neutrophilic dermatoses.

[0531] Kits

[0532] Also provided by the present disclosure is a kit or article of manufacture comprising an antibody that inhibits G-CSF signaling packaged with instructions for use in a method described herein.

[0533] In some examples, the kit or article of manufacture comprises (a) a container comprising an antibody 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 a neutrophilic skin condition in a subject as described herein.

[0534] 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 for treating the neutrophilic skin 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 antibody that inhibits G-CSF signaling. The label or package insert indicates that the composition is used for treating the neutrophilic skin condition, with specific guidance regarding dosing amounts and intervals of antibody 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.

[0535] Alternatively or additionally, the kit or article of manufacture may comprise a subcutaneous administration device, which delivers to a subject a fixed dose of 25 mg or 50 mg of the antibody that inhibits G-CSF signaling. Devices suitable for subcutaneous delivery include: a syringe (including a pre-filled syringe); an injection device (e.g. the INJECT - EASE™ and GENJECT™ device); an infusion pump (such as e.g. Accu-Chek™); an injector pen (such as the GENPEN™); a needleless device (e.g. MEDDECTOR™ and BIOJECTOR™); an autoinjector, a subcutaneous patch delivery system, etc.

[0536] It will be appreciated by persons skilled in the art that numerous variations and / or modifications may be made to the above-described embodiments, without departing from the broad general scope of the present disclosure. The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive.

[0537] The present disclosure includes the following non-limiting Examples.

[0538] EXAMPLES

[0539] 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)

[0540] Objectives

[0541] Primary Objective: To evaluate the safety of repeat doses of intravenous (IV) CSL324 in subjects with hidradenitis suppurativa (HS) and palmoplantar pustulosis (PPP).

[0542] Secondary Objectives:

[0543] 1. To determine the pharmacokinetics (PK) of repeat doses of IV CSL324

[0544] 2. To investigate the immunogenicity of repeat doses of IV CSL324

[0545] Exploratory Objectives

[0546] 1. To measure the effect of CSL324 on the inflammatory cell infiltrate, including neutrophilic infiltration, in skin lesion biopsies

[0547] 2. To explore the pharmacodynamic (PD) properties and potential biomarkers of CSL324

[0548] 3. To assess the potential efficacy, including patient-reported outcomes, of repeat doses of IV CSL324

[0549] 4. To explore potential PK / PD / efficacy relationships Methodology

[0550] 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.

[0551] 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.

[0552] 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.

[0553] Number of Subjects

[0554] Planned: 40 subjects total (20 subjects per indication).

[0555] Actual: 39 subjects total (22 subjects with HS and 17 subjects with PPP).

[0556] 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.

[0557] Investigational Product, Dose and Mode of Administration, Batch Number(s)

[0558] 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).

[0559] 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.

[0560] Duration of Treatment:

[0561] The duration of the treatment for each subject was 15 weeks, followed by a 9-week Follow-up Period.

[0562] Criteria for Evaluation

[0563] Primary Endpoint:

[0564] 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.

[0565] Secondary Endpoints:

[0566] The secondary endpoints for this study were as follows:

[0567] • 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)

[0568] • Serum PK parameters of CSL324 for the last dose administered including the following:

[0569] O Cmax

[0570] O Tmax

[0571] 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

[0572] • Trough concentration of CSL324 for each dose of CSL324 administered

[0573] • 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)

[0574] • Presence of anti-CSL324 antibodies in serum

[0575] Exploratory Endpoints:

[0576] The exploratory endpoints for this study were as follows:

[0577] • 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

[0578] • Serial ANC profile in whole blood

[0579] • 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)

[0580] • Genetic profile from peripheral whole blood (optional) was analyzed and reported separately

[0581] • Transcriptome profile, including the neutrophil signature, from peripheral whole blood was analyzed and reported separately

[0582] • Neutrophil profile shift in peripheral blood films

[0583] • Efficacy, including patient-reported outcome measures: o All subjects:

[0584] ■ Dermatology Life Quality Index (DLQI) score o Subjects with HS only:

[0585] ■ Hidradenitis Suppurativa Clinical Response (HiSCR)

[0586] ■ International Hidradenitis Suppurativa Severity Score System (IHS4) ■ HS Physician Global Assessment (HS-PGA) score

[0587] ■ Pain numerical rating scale (NRS) score o Subjects with PPP only:

[0588] ■ ppPASI score

[0589] ■ PPP Physician Global Assessment (PPP-PGA) score

[0590] • PK / PD / efficacy relationships with appropriate biomarkers were explored and reported separately.

[0591] Statistical Methods

[0592] The sample size for this study was based upon empirical considerations; no formal sample size calculation was performed.

[0593] Analysis Populations:

[0594] 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.

[0595] The Safety Analysis Set comprised all subjects in the Enrolled Analysis Set who received at least 1 partial dose of CSL324.

[0596] 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.

[0597] 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.

[0598] 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.

[0599] 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.

[0600] Data Handling Conventions:

[0601] 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.

[0602] Changes to subjects’ visits caused by the coronavirus disease 2019 (COVID- 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 CO VID- 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.

[0603] Partially or completely missing dates were not imputed.

[0604] Pharmacokinetic and Pharmacodynamic Analyses:

[0605] 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.

[0606] Biomarker parameters were summarized by treatment group and study indication for the PD Analysis Set by descriptive statistics.

[0607] 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.

[0608] Efficacy Analyses:

[0609] Efficacy parameters were summarized using descriptive statistics:

[0610] • 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

[0611] • For subjects with PPP: ppPASI score

[0612] • For both subjects with HS and PPP: disease-specific PGA and DLQI (total and subscores)

[0613] Safety Analyses:

[0614] All safety analyses were based on the Safety Analysis Set, and summaries included all scheduled safety assessments. The extent of exposure to CSL324 was summarized by treatment group and study indication using descriptive statistics.

[0615] 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.

[0616] 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 CO VID- 19-associated AEs were included in standard AE tables. Separate summaries of CO VID- 19-associated AEs were also produced in the same way as for all AEs.

[0617] 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.

[0618] 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.

[0619] Results

[0620] Subject Disposition:

[0621] 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.

[0622] Protocol Deviations:

[0623] 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.

[0624] Demographics and Other Baseline Characteristics:

[0625] 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.

[0626] 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.

[0627] Overall, 39 subjects (100.0%) recorded intake of at least 1 concomitant medication.

[0628] Pharmacokinetics Results:

[0629] • 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.

[0630] • 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. The serum PK parameters of CSL324 were generally similar between subjects with PPP and HS in both cohorts.

[0631] • 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. Pharmacodynamics Results:

[0632] • 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.

[0633] • 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. • 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.

[0634] • 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.

[0635] • 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.

[0636] Efficacy Results:

[0637] • Efficacy assessments were exploratory endpoints in this study. The following trends in clinical responses were observed.

[0638] • 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).

[0639] • 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).

[0640] Safety Results:

[0641] • Overall, 33 subjects (84.6%) experienced 198 TEAEs, of which 70 TEAEs in 16 subjects (41.0%) were related to CSL324.

[0642] • 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.

[0643] • 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.

[0644] • 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.

[0645] • 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.

[0646] • 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).

[0647] • 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.

[0648] • 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).

[0649] • Two subjects (5.1%) experienced 2 TEAEs (moderate Neutropenia and moderate Hypersensitivity) that led to permanent discontinuation of CSL324 and withdrawal from the study. • 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.

[0650] • One subject had treatment-emergent antidrug antibodies at a low titer of 20 at 2 timepoints, which resolved on subsequent visits.

[0651] Conclusions

[0652] 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.

[0653] 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). 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)

[0654] Objectives

[0655] Primary Objectives:

[0656] 1. To characterize the pharmacokinetics (PK) of CSL324 in Japanese and White subjects following a single subcutaneous (SC) administration of CSL324.

[0657] 2. To compare the PK of CSL324 in Japanese and White subjects following a single SC administration of CSL324.

[0658] Secondary Objectives:

[0659] 1. Evaluate the safety and tolerability of a single SC dose of CSL324 in Japanese and White subjects.

[0660] 2. Further characterize the PK properties of CSL324 in healthy Japanese and White subjects.

[0661] 3. Investigate the immunogenicity of a single SC dose of CSL324.

[0662] Exploratory Objective: To assess the pharmacodynamic (PD) properties of CSL324 after a single SC dose of CSL324 in Japanese and White subjects.

[0663] Methodology

[0664] 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.

[0665] 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 followup visits and the End of Study Visit for further blood sampling and safety assessments.

[0666] 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.

[0667] 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.

[0668] Safety, Pharmacokinetic, and Pharmacodynamic Assessments

[0669] 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.

[0670] 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.

[0671] Number of Subjects

[0672] Planned: minimum of 32 and maximum of approximately 48.

[0673] Actual: 32 (Cohort 1: 16; Cohort 2: 16).

[0674] Diagnosis and Main Criteria for Inclusion

[0675] 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.

[0676] 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.

[0677] Investigational Product, Dose and Mode of Administration, Batch Number(s)

[0678] 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:

[0679] • Cohort 1 : 0.3 mg / kg CSL324.

[0680] • Cohort 2: 0.8 mg / kg CSL324.

[0681] The volume administered was determined by the subject’s dose cohort and weight on Day -1.

[0682] 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.

[0683] Batch numbers for CSL324: 27212AC49A; 27212AC49B; 27212AC49C.

[0684] Batch numbers for placebo: 27213AC43A; 27213AC43B; 27213AC43C.

[0685] Duration of Treatment

[0686] Subjects received CSL324 or placebo as a single dose on Day 1 and were followed up until Day 56.

[0687] Criteria for Evaluation

[0688] Primary Endpoints:

[0689] • 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): o Mean (standard deviation [SD]) and geometric mean (geometric % coefficient of variation [CV]). o 90% confidence interval (CI) for the ratio of geometric means using White subjects as the reference.

[0690] Secondary Endpoints:

[0691] • TEAEs up to Day 56.

[0692] • AEs localized to the administration site up to Day 7.

[0693] • PK parameters: o Time to reach C max (T max). o Terminal half-life (ti / 2). o Apparent clearance (CL / F). o Apparent volume of distribution (Vz / F).

[0694] • Presence of anti-CSL324 antibodies in serum on Day 1 before IP administration and on Days 28 and 56 after IP administration.

[0695] Statistical Methods

[0696] Analysis Populations

[0697] 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.

[0698] The Safety Analysis Set comprised all subjects in the Enrolled Analysis Set who received at least 1 partial dose of IP.

[0699] 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.

[0700] 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.

[0701] General Considerations

[0702] 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.

[0703] Pharmacokinetic Analyses

[0704] 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.

[0705] The Cis were then exponentiated to form 90% Cis for the ratios of the geometric means in the original scale.

[0706] Pharmacodynamic Analyses (Biomarker Analyses)

[0707] 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.

[0708] 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.

[0709] PK / PD Analyses

[0710] 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.

[0711] 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.

[0712] Results

[0713] Subject Disposition

[0714] 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.

[0715] 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. Overall, 31 subjects (96.9%) completed the study. One subject (placebo-treated) in Cohort 2B was discontinued due to Other reasons.

[0716] Demographics

[0717] 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.

[0718] Pharmacokinetics

[0719] The serum CSL324 concentrations in Cohort 1 (0.3 mg / kg) were variable (%CV

[0720] > 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.

[0721] 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 Results:

[0722] Subject Disposition

[0723] 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.

[0724] 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.

[0725] Overall, 31 subjects (96.9%) completed the study. One subject (placebo-treated) in Cohort 2B was discontinued due to Other reasons.

[0726] Demographics

[0727] 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.

[0728] Pharmacokinetics

[0729] The serum CSL324 concentrations in Cohort 1 (0.3 mg / kg) were variable (%CV

[0730] > 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.

[0731] 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%).

[0732] 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.

[0733] 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%).

[0734] 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.

[0735] 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.

[0736] 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.

[0737] Pharmacodynamics

[0738] 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.

[0739] 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.

[0740] Safety

[0741] 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 whomreceived 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%]).

[0742] 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.

[0743] 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.

[0744] 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.

[0745] 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.

[0746] 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.

[0747] No subjects experienced Grade 3 or 4 infection and there was no evidence of an increased susceptibility to infection noted in this study.

[0748] There were no subjects with presence of anti-CSL324 antibodies in serum over the treatment period.

[0749] There were no deaths or SAEs in this study. No subject discontinued / withdrew from the study due to TEAEs.

[0750] Conclusions

[0751] 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.

[0752] 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. Example 3: G-CSF gene signature present in adalimumab-failed HS patients

[0753] The inventors derived a gene signature of G-CSF activity using the transcriptomic data generated from peripheral samples collected from the first in human clinical trial with CSL324, the CSL324_1001 (ACTRN 12616000846426), 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. 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 (postfilgrastim / G-CSF) vs PreTreatment 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.

[0754] G-CSF gene signature is elevated in skin from HS patients, and it remains elevated in adalimumab non-responders but not in responders

[0755] The inventors evaluated the gene expression of the G-CSF gene signature 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 double-blind, 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).

[0756] The inventors examined the expression levels of genes in the G-CSF gene signature within lesional skin samples from the adalimumab trial. This analysis was conducted through sample-level enrichment analysis using the R package GSVA (hdps; / / gith b.coinAcasieio / GS VA) (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.

[0757] The G-CSF gene signature exhibits 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 is no longer enriched in responders (depicted by black dots, HS lesional_POST) but remains elevated in non-responders (illustrated by white dots, HS lesional_POST). This observation 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.

[0758] Thl7 / IL-17 gene-sets did not segregate adalimumab non-responders from responders

[0759] The inventors also investigated the activation of Thl7 / IL-17 pathways 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.

[0760] 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 Thl7 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 / gsea / msigdb / ).

[0761] 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 below 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).

[0762] Top disease-associated pathways enriched in HS lesions do not segregate adalimumab nonresponders from responders

[0763] The inventors then evaluated whether other key processes altered in the HS samples 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 inventors compared 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 by performing fast ranked Gene Set Enrichment Analysis (fGSEA) using the R package ClusterProfiler (http : / / bloconductor.org / packages / relea eAsioc / htmVc1usterProfsler.htm) (Yu et al., 2012). fGSEA is used to assess if certain groups of genes that work together in specific functions or pathways are more or less active in different conditions, like in this case healthy and HS disease state.

[0764] Figure 4 below shows the top 10 pathways identified as differential regulated in skin samples from HS patients compared to healthy skin. The pathways are 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.

[0765] The inventors then assess the enrichment score of the top 3 activated pathways identified in the HS patients in the skin samples obtained post-adalimumab treatment. As shown in Figure 5, as expected 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-responders 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.

[0766] Altogether, this data provides strong scientific rationale supporting the potential for CSL324 to have clinical benefit in HS adalimumab non-responders

[0767] Example 4: Fixed dosing modelling data

[0768] The purpose of the analysis performed by the inventors in this Example was to develop population pharmacokinetic (PK) and absolute neutrophil count (ANC) response models to characterize the concentration-time profile of CSL324 and the effect of CSL324 on ANC in healthy subjects and patients with neutrophilic skin conditions such as HS and PPP. The population PK and ANC models were then used to determine an optimal fixed dose regimen for treating neutrophilic skin conditions.

[0769] The modelling analysis included pooled CSL324 serum concentration-time data (1120 observations; 87 subjects) and ANC data (1880 observations; 87 subjects) from three clinical studies, including two Phase 1 studies in healthy subjects (25 healthy subjects from Study CSL324_1001 and 23 healthy subjects [12 Caucasian; 11 Japanese] from Study CSL324_1003) and one Phase lb study in patients with HS or PPP (22 HS patients and 17 PPP patients from Study CSL324_1002, i.e., the study described in Example 1). Healthy subjects received single dose placebo or 0.1 to 1 mg / kg CSL324 via intravenous (IV) infusion (CSL324_1001) or 0.3 and 0.8 mg / kg via subcutaneous (SC) injection (CSL324_1003), or multiple dose CSL3240.6 mg / kg IV every 3 weeks (Q3W) x 3 doses (CSL324_1001); HS and PPP patients received 0.3 or 0.6 mg / kg CSL324 IV Q3W x 5 doses (CSL324_1002).

[0770] CSL324 PK model

[0771] Prior to PK model development, exploratory data analysis was performed to characterize the observed data and inform model selection. CSL324 concentrations were modeled in units of nanomoles per liter (nM) without transformation. A likelihood-based method that accounted for censoring of concentrations below the limit of quantification (BLQ) (M3 method) was used since 32.1% of all post-dose concentrations were BLQ in the pooled analysis dataset (23.8% to 37.4% BLQ for individual studies). First, a base model was developed with assessment of the random effects structure. Covariates were added to the PK base model using a stepwise forward inclusion procedure to establish the full covariate model followed by backward covariate elimination to identify a parsimonious final PK model. The final PK model was evaluated using a prediction-corrected visual predictive check (pcVPC) to determine predictive performance of the model.

[0772] The CSL324 concentration-time profile was best characterized by a population PK model with two-compartment disposition, parallel linear and non-linear (Michaelis-Menten) elimination, and first order absorption following SC administration including a lag time for absorption. Inter-individual variability was estimated for clearance (CL), volume of the central compartment (Vc), volume of the peripheral compartment (Vp), and the SC absorption rate constant (Ka), and a combined additive and proportional error model was used to describe residual variability. Additive error was estimated separately for healthy subjects and patients (HS and PPP patients in Study CSL324_1002, i.e., the study described in Example 1). Known effects of body weight on both CL and Vc of monoclonal antibodies were evaluated and found to be statistically significant for CSL324 in the final model. Covariate effects of patient status (HS [N=22] or PPP [N= 17] vs. healthy subjects [N=48]) on CL, sex (female [N=50] vs. male [N=37]) on CL and race (Asian [N=12] vs. non- Asian [N=75]) on Ka were not significant.

[0773] The model-estimated exponent for the weight relationship with CL was higher than expected (2.21). Moreover, a potential confounding effect of patient status with baseline body weight was noted, as the upper end of the baseline body weight range (>100 kg) included only HS and PPP patients while all healthy subjects had baseline body weights <100 kg. Therefore, in the updated final model, exponents for CL and Vc were fixed to the average theoretical exponents derived from studies of monoclonal antibodies (0.583 for weight on CL and 0.511 for weight on Vc, see Hendrikx et al. The Oncologist 2017, 22: 1212). Diagnostic plots and model evaluation using pcVPC demonstrated adequate predictive performance of the updated final model. Typical PK parameters from the updated final model were as follows for a 75.2 kg subject: linear CL of 0.00743 L / hr (97.7% CV); Q of 0.0268 L / hr; Vc of 2.98 L (17.7% CV), Vp of 1.93 L (47.7% CV), Vmax of 0.361 nmol / hr, Km fixed at 1.37 nM (based on the binding affinity of CSL324 to human colony- stimulating factor receptors), Ka of 0.0078 hr1(83.9% CV), lag time of 1.36 hr, and SC bioavailability of 92%. At baseline (where concentration is 0 at time 0), the model estimated that nonlinear clearance comprises -97% of total clearance. As concentrations increase, the contribution of nonlinear clearance diminishes and linear clearance predominates, with nonlinear clearance accounting for 50% of total CL at -7 pg / mL and 20% of total CL at -28 pg / mL.

[0774] Model-based simulations were performed to predict CSL324 exposure metrics (AUC, Cmax, Cmin) with fixed SC doses of 25 mg, 50 mg, and 60 mg CSL324 every 2 weeks (Q2W) for virtual subjects with body weight ranging from 40-170 kg in 10-kg increments. The results for the 25 mg and 50 mg CSL324 Q2W SC dose regimens are shown in Figure 9. Median exposures in the lowest weight group (40 to 50 kg) and highest weight group (160 to 170 kg) were predicted to be within approximately + / - 50% of the predicted exposure for a reference body weight of 90 kg (median weight for HS and PPP patients in Study 1002). The 90% confidence intervals of predicted exposure metrics were largely overlapping across all weight groups.

[0775] ANC model

[0776] A model to characterize the effect of CSL324 exposure on ANC (as depicted in Figure 7) was developed by the inventors based on a model of chemotherapy-induced myelosuppression published by Friberg et al. (Friberg et al. Clin. Oncol. 2002, 20:4713). Individual PK parameters estimated from the updated final PK model were used in the ANC analysis. Natural log-transformed ANC values were modelled due to apparent log-normal distribution of the ANC data. The model was tested against observed data from the CSL324 1002 trial described in Example 1. Based on model diagnostics including VPCs (Figure 8), the model adequately described the drug-induced decline in ANC and return to baseline with drug elimination.

[0777] Simulations were performed to explore changes in ANC with flat SC-dose administration and the proportion of subjects predicted to experience neutropenia. The results are summarized for doses of 25 mg, 50 mg and 60 mg CSL324 Q2W SC in Table 2 below.

[0778] Table 2. Proportion of Subjects Predicted to Experience Neutropenia Under Various CSL324 SC Dosing Regimens at Week 16.

[0779] Predicted median ANC levels remained above the threshold of 1.5 xlO9cells / L for 16 weeks at 50 mg Q2W (and near the threshold from Week 16 to Week 32), and above the threshold for 32 weeks at 25 mg Q2W. Cumulative incidence of Grade 3 neutropenia was predicted to be <30% at doses up to 60 mg CSL324 Q2W SC for 16 weeks.

[0780] The following conclusions were drawn from the above analyses:

[0781] • A population PK model including two-compartment disposition, parallel linear and nonlinear (Michaelis-Menten) elimination, and first order absorption following SC administration including a lag time for absorption adequately described the concentrationtime profile of CSL324 after IV or SC administration in healthy subjects and patients with HS or PPP based on data from a previous trial (Example 1), chronic dermatological conditions characterized by neutrophilic infiltration.

[0782] • CSL324 exposure was higher with lower body weight; however, in simulations of flat SC dosing the exposures of all weight groups (ranging from 40-50 kg to 160-170 kg) were within approximately + / -50% of that of the 90 kg reference subject.

[0783] • The developed model for ANC with CSL324 exposure predicted that 8.3% 16%, and 17% of subjects would experience Grade 3 neutropenia after 16 weeks of treatment at doses of 25 mg, 50 mg, and 60 mg CSL324 Q2W SC, respectively. The model also predicted that 6.8%, 13%, and 13% of subjects would experience Grade 4 neutropenia after 16 weeks of treatment at doses of 25 mg, 50 mg, and 60 mg CSL324 Q2W SC, respectively. Lastly, 27%, 38%, and 43% of subjects were predicted to experience lower than Grade 3 neutropenia (i.e., Grades 1 or 2) after 16 weeks of treatment at doses of 25 mg, 50 mg, and 60 mg CSL324 Q2W SC, respectively.

[0784] • Performed model-based predictions of the relationship between CSL324 exposure and ANC indicated that the use of a Q2W SC dose regimen at doses of 25 or 50 mg was optimal, based on maximizing exposure to the antibody while minimizing risk of severe neutropenia. Example 5: A Randomized, Double-blind, Placebo-controlled, Phase 2 Study to Evaluate the Efficacy and Safety of Anumigilimab in Patients with Hidradenitis Suppurativa Who Have Had an Inadequate Response to Anti-TNF Therapy PROTOCOL SUMMARY

[0785] Synopsis

[0786] Schedule of Activities

[0787] Table 3. Treatment Period A.

[0788]

[0789]

[0790] ANC = absolute neutrophil count; DLQI = Dermatology Life Quality Index; ECG = electrocardiogram; EOS = end of study; EOT = end of treatment; FDA = Food and Drug Administration; HiSQOL = Hidradenitis Suppurativa Quality of Life; HS = hidradenitis suppurativa; IP = investigational product; IRT = interactive response technology; NRS = numerical rating scale; PD = pharmacodynamic(s); PK = pharmacokinetic(s); POC = point of care;

[0791] PRO = patient-reported outcomes; RNA = ribonucleic acid; UNS = unscheduled.

[0792] 5aIf a subject withdraws from treatment before completing Treatment Period A or is not willing to continue to Treatment Period B, the subject will not be (re- jrandomized, will undergo the assessments scheduled for the Day 225 (EOT) Visit, and will subsequently attend the Follow-up Visits (Day 252 and Day 280, 4 and 8 weeks after the last IP administration, respectively) (Section entitled “Treatment Period B”). If a subject withdraws or is withdrawn from study, the subject will undergo the assessments scheduled for the Day 280 (EOS) Visit (Section entitled “Treatment Period B”). b An unscheduled clinical encounter may occur at any time if the investigator believes an unscheduled visit is clinically warranted. Additional assessments or

[0793] 10 procedures may be performed as needed at the investigator’s discretion to follow unresolved findings of clinical concern. c If the subject opts for home health visits for this study day and it is available, the home health nurse will draw blood for hematology / ANC, PK, PD, research serum sample, and RNA whole blood sample, as appropriate. Adverse event and concomitant medication information may not be recorded by the home health nurse (unless authorized by the investigator) and will be recorded by the investigator or delegate at the next site visit. Testing for serum follicle-stimulating hormone may be performed to confirm postmenopausal status in female subjects aged 45 to 60 years (inclusive) with

[0794] 15 amenorrhea for > 1 year.

[0795] eThe predose ANC may be performed at a local laboratory or via the POC device. ANC results must be available to the investigator prior to dosing. Subjects with ANC value < 1 x 109cells / L will skip dose and will not be dosed until the ANC value is > 1 x 109cells / L. Please refer to stopping rules in the section entitled “Discontinuation of Investigational Product” below for details. If the local laboratory cannot provide ANC results within approximately 4 hours after the blood draw on the day of dosing, then the subject will be required to provide a blood sample up to 48 hours before dosing to ensure that ANC results (either from a central or local laboratory) are available

[0796] 5 prior to dosing. Blood draw for ANC assessment obtained prior to dosing days may utilize home health visits, if available. ANC may also be assessed by a POC finger prick test using an FDA-cleared device in select clinical sites, when available. f

[0797] One PK, PD, and hematology specimen to be taken on Day 3, 5, or 7 and 1 PK, PD, and hematology specimen to be taken on Day 101, 103, or 105 based on subject convenience in attending site visits. On days when IP is administered, PK, PD, hematology, research serum, and RNA samples will be collected predose, and on days when IP is not administered, PK, PD, hematology, research serum, and RNA samples can be collected, if possible, at approximately the same time of the day as on other

[0798] 10 days.

[0799] § PROs should be completed ahead of other study procedures at each study visit. Blood samples can be drawn earlier to ensure availability of ANC results prior to dosing. h If ANC values drop below 1 x 109cells / L, refer to the stopping rules and guidance on resuming administration of IP.

[0800] 1IP to be given after all study assessments are completed.

[0801] 15 J Ultrasound imaging may be used to identify the presence or absence of a dermal tunnel or any pus collection beneath the skin surface in a subset of study sites.

[0802] 20

[0803] 25

[0804] 30

[0805] Table 4. Treatment Period B.

[0806]

[0807] ANC = absolute neutrophil count; DLQI = Dermatology Life Quality Index; ECG = electrocardiogram; EOS = end of study; EOT = end of treatment; FDA = Food and Drug Administration; HiSQOL = Hidradenitis Suppurativa Quality of Life; HS = hidradenitis suppurativa; IP = investigational product; IRT = interactive response technology; NRS = numerical rating scale; PK = pharmacokinetic(s); POC = point of care; UNS = unscheduled.

[0808] 5aIf a subject withdraws from treatment before completing Treatment Period B, the subject will undergo the assessments scheduled for the Day 225 (EOT) Visit and will subsequently attend the Follow-up Visits (Day 252 and Day 280, 4 and 8 weeks after the last IP administration, respectively). If a subject withdraws or is withdrawn from study, the subject will undergo the assessments scheduled for the Day 280 (EOS) Visit. b An unscheduled clinical encounter may occur at any time if the investigator believes an unscheduled visit is clinically warranted. Additional assessments or procedures may be performed as needed at the investigator’s discretion to follow unresolved findings of clinical concern.

[0809] 10cIf the patient opts for home health visits for this study day and it is available, the home health nurse will draw blood for hematology / ANC, PK, and research serum sample, as appropriate. Adverse event and concomitant medication information may not be recorded by the home health nurse (unless authorized by the investigator) and will be recorded by the investigator or delegate at the next site visit. The predose ANC may be performed at a local hematology laboratory or via the POC device. ANC results must be available to the investigator prior to dosing.

[0810] Subjects with ANC value < 1 x 109cells / L will skip dose and will not be dosed until the ANC value is > 1x109cells / L. Please refer to stopping rules in the section

[0811] 15 entitled “Discontinuation of Investigational Product” below for details. If the local laboratory cannot provide ANC results within approximately 4 hours after the blood draw on the day of dosing, then the subject will be required to provide a blood sample up to 48 hours before dosing to ensure that ANC results (either from a central or local laboratory) are available prior to dosing. Blood draw for ANC assessment obtained prior to dosing days may utilize home visits, if available. ANC may also be assessed by a POC finger prick test using an FDA-cleared device in select clinical sites when available. e One PK and hematology sample to be taken on Day 115, 117, or 119, and 1 PK and hematology sample to be taken on Day 213, 215, or 217 based on subject

[0812] 20 convenience in attending site visits. On days when IP is administered, PK, research serum, and hematology samples will be collected predose, and on days when IP is not administered, PK, hematology, and research serum samples will be collected, if possible, at approximately the same time of the day as on other days. f

[0813] PROs should be completed ahead of other study procedures at each study visit. Blood samples can be drawn earlier to ensure availability of ANC results prior to dosing.

[0814] § If ANC values drop below 1 x 109cells / L, refer to the stopping rules and guidance on resuming administration of IP.

[0815] 25 h IP is to be given after all study assessments are completed

[0816] INTRODUCTION

[0817] Study Rationale

[0818] The safety and pharmacokinetics (PK) of intravenously (IV) administered anumigilimab in subjects with moderate to severe HS was assessed as favorable in an open-label, phase lb study (CSL324_1002). This study also demonstrated dose- dependent suppression of neutrophil abundance and preliminary clinical responses when anumigilimab was administered at 2 different dose levels. The aim of this phase 2 study is to characterize the clinical efficacy, safety, and PK and pharmacodynamic (PD) effects of repeat doses of subcutaneously (SC) administered anumigilimab in subjects with HS. Adalimumab (anti- TNF) is an approved biologic therapy for patients with moderate to severe HS, and there are currently no approved therapies for patients who do not respond or have lost response to adalimumab treatment. Hence, the unmet need is most acute in this population, and this study will evaluate efficacy of anumigilimab in subjects with moderate to severe HS who have had an inadequate response to anti-TNF therapy.

[0819] Benefit / Risk Assessment

[0820] Anumigilimab has been administered to 102 subjects (63 healthy subjects and 39 subjects with HS or palmoplantar pustulosis [PPP]), providing an early understanding of the risks and benefits associated with anumigilimab treatment.

[0821] More detailed information about the risks and mitigation strategies may be found in Table 5 below.

[0822] Benefit Assessment

[0823] The phase lb study CSL324_1002 showed preliminary signs of disease improvement in both HS and PPP subjects when given repeat doses of IV anumigilimab. These improvements included a reduced disease score, such as the International Hidradenitis Suppurativa Severity Score System (IHS4) and the Palmoplantar Pustulosis Area and Severity Index.

[0824] The benefits of anumigilimab administration for subjects with HS will be further evaluated and confirmed. Treatment will be considered to have the highest impact if anumigilimab is able to reduce disease severity, decrease the occurrence of flares, and improve quality of life by controlling pain and discharge in patients with moderate to severe HS who have had an inadequate response to anti-TNF therapy. Risk Assessment

[0825] Table 5. Risks and Mitigation Strategies.

[0826] ADA = antidrug antibody; AE = adverse event; AESI = adverse event of special interest; ANC = absolute neutrophil count; EOS = End of Study; HS = hidradenitis suppurativa; IV = intravenous (ly); mAb = monoclonal antibody; PPP = palmoplantar pustulosis; SC = subcutaneous(ly).

[0827] Benefit / Risk Conclusion

[0828] The potential risks associated with anumigilimab administration are adequately mitigated with the measures taken to minimize risks to the subjects participating in this study (ie, predefined inclusion / exclusion criteria, halting criteria, monitoring of adverse events of special interest [AESIs], frequent ANC assessment, and regular medical monitoring).

[0829] The benefit / risk assessment of anumigilimab supports the further exploration of anumigilimab in patients with moderate to severe HS who have had an inadequate response to anti-TNF therapy, and it will continue to be evaluated as the clinical program advances.

[0830] OBJECTIVES AND ENDPOINTS

[0831] Primary Objective and Endpoint

[0832] Primary Objective

[0833] The primary objective of the study is to evaluate the efficacy of anumigilimab administered SC in Treatment Period A to subjects with moderate to severe HS who have had an inadequate response to anti-TNF therapy. The primary end point is summarised in Table 6 below.

[0834] Table 6. Primary Endpoint.

[0835] IHS4 = International Hidradenitis Suppurativa Severity Score System. I l l

[0836] Primary Estimand Description

[0837] The primary interest is to assess the efficacy of anumigilimab during treatment from Day 1 through Week 16, controlling for the potential confounding of subjects who withdrew from treatment or the study before the end of Week 16 or had an intake of prohibited medication.

[0838] The primary estimand in line with the primary interest of the study is described as follows:

[0839] • Population: The target subject population defined by eligibility criteria and who were included in the modified intention-to-treat (mITT).

[0840] • Variable: Absolute change from baseline of IHS4 at the end of Week 16.

[0841] • Intercurrent events (ICE): Permanent withdrawal from the investigational product (IP) or the study or intake of prohibited medication. Measures after these ICEs will not be used for analysis.

[0842] Secondary Objectives and Endpoints

[0843] Secondary Objectives

[0844] The secondary objectives of the study are as follows:

[0845] 1. To assess the safety of anumigilimab administered SC to subjects in this study

[0846] 2. To assess additional measures of efficacy of anumigilimab administered SC in Treatment Period A to subjects in this study

[0847] 3. To assess the immunogenicity of repeat doses of anumigilimab administered SC to subjects in this study.

[0848] Secondary endpoints are summarised in Table 7 below.

[0849] Table 7. Secondary Endpoints.

[0850] ADA = antidrug antibody; AE = adverse event; AESI = adverse event of special interest; AN = abscess and nodule; DLQI = Dermatology Life Quality Index; HiSCR50 = > 50% reduction from baseline in the total abscess and nodule count with no increase in abscess or draining tunnel count; HiSCR75 = > 75% reduction from baseline in the total abscess and nodule count, with no increase in abscess or draining tunnel count; IHS4- 55 = > 55% decrease from baseline in International Hidradenitis Suppurativa Severity Score System score; mHISCR = modified Hidradenitis Suppurativa Clinical Response; NRS = numerical rating scale; SAE = serious adverse event; TEAE = treatment-emergent adverse event.

[0851] Exploratory Objectives and Endpoints

[0852] Exploratory Objectives

[0853] The exploratory objectives of this study are as follows:

[0854] 1. To assess exploratory efficacy measures in Treatment Period A

[0855] 2. To further assess safety in Treatment Period A and in Treatment Period B 3. To determine the PK and PD (including biomarkers) of repeat doses of SC anumigilimab in subjects in this study

[0856] 4. To assess efficacy in Treatment Period B only and in the combined overall Treatment Period (A and B) 5. To assess changes in biomarkers in blood and skin, including transcriptomic analysis.

[0857] Exploratory endpoints are summarised in Table 8 below. Table 8. Exploratory Endpoints.

[0858] AN = abscess and nodule; ANC = absolute neutrophil count; ANdT = abscess, nodule, and draining tunnel;

[0859] AUCo-iast = area under the concentration-time curve from time 0 to the last measurable concentration;

[0860] AUCtau.ss = area under the concentration-time curve within a dosing interval at steady state; CL / F = apparent clearance; Cmax,i = maximum concentration after first dose; Cmax,ss = maximum concentration at steady state; ECG = electrocardiogram; G-CSF = granulocyte colony-stimulating factor; HiSCR50 = > 50% reduction from baseline in the total abscess and nodule count with no increase in abscess or draining tunnel count; HiSCR75 = > 75% reduction from baseline in the total abscess and nodule count, with no increase in abscess or draining tunnel count; HiSQOL = Hidradenitis Suppurativa Quality of Life; HS = hidradenitis suppurativa; IHC = immunohistochemical; IHS4 = International Hidradenitis Suppurativa Severity Score System; mHISCR = modified Hidradenitis Suppurativa Clinical Response; PD = pharmacodynamic(s); PK = pharmacokinetic(s); SC = subcutaneous; Vz / F = apparent volume of distribution.aWhere applicable, endpoints will be summarized by visit. All endpoints will be presented by treatment (anumigilimab or placebo for Treatment Period A; anumigilimab-low dose, anumigilimab -high dose, placebo-low dose, or placebo- high dose for Treatment Period B). For Treatment Period A, differences between treatments will be presented.

[0861] STUDY DESIGN

[0862] Overall Design

[0863] This is a multicenter, randomized, double-blind, placebo-controlled phase 2 study investigating the efficacy and safety of repeat SC doses of anumigilimab in subjects with HS. A total of approximately 96 subjects, between >18 and < 75 years of age, are planned to be enrolled in this study. The study will be conducted in sites in the USA, Canada, Germany, Australia, and Denmark. Additional countries may be added. Subjects with moderate to severe HS per IHS4 guidelines (ie, IHS4 > 4) and who have demonstrated inadequate response to anti-TNF therapy will be eligible for enrolment.

[0864] 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 (Figure 6).

[0865] Screening Period

[0866] Subjects who do not meet the criteria for participation in this study (screen failure) may be rescreened once. The retesting of abnormal screening tests, which are exclusionary and considered potentially erroneous, is allowed at an unscheduled visit during the Screening Period (to reassess eligibility). This may not be considered rescreening. Further details are provided in the section entitled “Screen Failure” below.

[0867] The study procedures to be performed during the Screening Period are provided in Table 3 above.

[0868] Subjects are considered “enrolled” in the study when they, or their legally acceptable representative, has agreed to participate in the clinical study after completing the informed consent process and screening. Potential subjects who are screened to determine their eligibility but fail screening or do not meet all eligibility criteria are not considered enrolled.

[0869] Treatment Period A

[0870] Eligible subjects will be randomized in a ratio of 2: 1 to the anumigilimab 50 mg SC every 2 weeks (Q2W) or placebo arms by means of an interactive response technology (IRT) system using the baseline IHS4 score as the stratification factor (see the section entitled “Randomization” below). Subjects will receive 50 mg of anumigilimab or placebo SC Q2W for 16 weeks.

[0871] Subjects will report to the study sites for IP administration; assessments of efficacy, safety, PK, PD / disease biomarkers, and immunogenicity; and collection of samples for research and RNA sequencing, as specified in Table 3 above. On days when the IP is administered, serum samples for PK, PD / disease biomarkers, safety laboratory, research, and RNA sequencing will be collected predose. On days when the IP is not administered, these samples can be collected at approximately the same time of the day as on previous days. Where available and authorized, blood draws for the visits indicated in Table 3 above can be performed by a qualified home health nurse.

[0872] 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]) at time points specified in Table 3 above. 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.

[0873] At the discretion of the sponsor, an optional interim analysis to assess nonbinding futility may be conducted after about 42% of subjects have completed Treatment Period A.

[0874] At the end of Treatment Period A, on Day 112, completers from each arm of Treatment Period A will be (re-)randomized in Treatment Period B in a ratio of 1 : 1 to either anumigilimab 50 mg SC Q2W or 25 mg SC Q2W.

[0875] The visit schedule for all subjects in Treatment Period A is provided in Table 3 above.

[0876] Skin Biopsy Substudy

[0877] 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]) according to the procedures in the section entitled “Pharmacodynamics and Biomarkers” below. 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 (eg, TNF signaling and IL- 17 signaling).

[0878] Treatment Period B

[0879] Subjects in Treatment Period B will receive either anumigilimab 50 mg SC Q2W or 25 mg SC Q2W for 16 weeks.

[0880] Subjects will report to study sites for anumigilimab administration; assessments of efficacy, safety, PK, and immunogenicity; and collection of samples for research, as specified in Table 4 above. On days when the IP is administered, all scheduled assessments, serum samples for PK, safety laboratory, and research serum samples will be collected predose. On days when the IP is not administered, these samples can be collected at approximately the same time of the day as on previous days. Where available and authorized, blood draws for the visits indicated in Table 4 above can be performed by a qualified home health nurse.

[0881] Efficacy assessments (assessment of HS lesions by the blinded examiner and algorithmic derivatives [IHS4, HiSCR50, HiSCR75, mHiSCR, and IHS4-55]), recording of concomitant therapies, and monitoring for AEs will be done for the duration of subject participation in Treatment Period B.

[0882] The visit schedule for subjects in Treatment Period B is provided in Table 4 above.

[0883] Follow-up Period

[0884] Subjects who complete Treatment Period B and subjects who withdraw from treatment early will enter the 8-week Follow-up Period for continued assessments of efficacy, safety, PK, and immunogenicity, as specified in Table 4 above.

[0885] The visit schedule for subjects in the Follow-up Period is provided in Table 4 above.

[0886] Scientific Rationale for Studv Design

[0887] Study CSL324_2001 is designed to extend the clinical experience with anumigilimab and evaluate the safety and efficacy of anumigilimab administered SC to subjects with moderate to severe HS who have had an inadequate response to anti-TNF therapy. The study will evaluate a single dose level of anumigilimab against placebo- treated subjects. The blinded study design will allow for unbiased assessment of safety and efficacy. The study will employ a blinded examiner to assess efficacy variables that require clinical judgment to minimize bias that may arise if changes in ANC values are visible to the examiner. Treatment Period B is included in the study to evaluate the safety of longer- term treatment with anumigilimab and the durability of response with different maintenance dosing regimens (50 or 25 mg administered SC Q2W). The study design also allows the evaluation of the safety, PK, and PD of a second dose (25 mg administered SC Q2W) of anumigilimab in Treatment Period B. Study CSL324_2001 will investigate the safety profile of anumigilimab, including the risk of neutropenia and infection. Skin biopsies, guided by ultrasound imaging to locate the presence of dermal tunnels or any pus collection, may be collected from a subset of up to 30 subjects in sites that have the expertise and equipment. Exploratory objectives have been included to better understand efficacy (including patient-reported outcomes), PK, and PD of SC administered anumigilimab in subjects with moderate to severe HS who have had an inadequate response to anti-TNF therapy.

[0888] Dose Rationale

[0889] The dose to be tested in this study is 50 mg of anumigilimab or placebo Q2W (2: 1 randomization), administered SC for a total of 16 weeks (blinded Treatment Period A). Following that, subjects in the 50 mg anumigilimab or placebo arm will either receive a 25 mg SC or 50 mg SC anumigilimab Q2W for an additional 16 weeks in the blinded Treatment Period B. Dose selection for the Study CSE324_2001 is based primarily on the inventors’ predictive modelling described in Example 4 and PK and PD (ANC) data obtained in the phase 1 studies, CSE324_1001, CSE324_1002, and CSE324_1003, which are described in Table 9 below.

[0890] Table 9. CSL Phase I studies

[0891] Anumigilimab demonstrated nonlinear PK after single doses of 0.1 mg / kg to 1 mg / kg administered IV, with exposure (area under the concentration-time curve from time 0 to infinity [AUCo-inf]) increasing in a more than dose-proportional manner. Minimal to no accumulation of anumigilimab was observed after 3 doses of 0.6 mg / kg administered IV every 3 weeks, with a mean (standard deviation [SD]) terminal elimination half-life (T1 / ?) of 251 (55.2) hours. Following single or multiple doses of anumigilimab, G-CSF receptor occupancy occurred rapidly and reached approximately 100% occupancy, even at the lowest dose tested (0.1 mg / kg), and was sustained at this level for longer periods of time as the dose increased.

[0892] In Study CSE324_1001, transient moderate neutropenia (ANC of < 1.0 to > 0.5 x 109cells / E) was observed in 1 subject (1 event) following a single dose of 1 mg / kg and in

[0893] 4 subjects (7 events) following 3 repeated doses of 0.6 mg / kg. In Study CSE324_1003, 1

[0894] Japanese subject experienced moderate neutropenia following a single dose of 0.8 mg / kg of anumigilimab administered SC.

[0895] In the phase lb study CSE324_1002 in subjects with HS, where 2 IV dose levels of anumigilimab were investigated, both dose levels of 0.3 mg / kg and 0.6 mg / kg exhibited the desired pharmacological effects and signs of target engagement. Even though clinical responses, as measured by either HiSCR50 / HiSCR75 or IHS4, were numerically better for the 0.3 mg / kg dose group compared to the 0.6 mg / kg dose group, clinically meaningful effects were observed at both doses. Also, dose-dependent increases in drug exposure and effects on neutrophils were observed.

[0896] Study CSE324_1002 was conducted with every 3 weeks (Q3W) dosing, and data show that trough concentrations (Ctrough) are mostly below the lower limit of quantitation, suggesting some period of drug holiday between doses. This further suggests that the T1 / ? of 251 hours after 3 doses of 0.6 mg / kg IV given 3 weeks apart in Study CSE 324_1001 is likely an over-estimation. Additionally, mean T* after an SC dose of 0.8 mg / kg is 124 hours, with a range of 65 to 196 hours, which is shorter than a comparable IV dose. Hence, Q2W dosing is appropriate from a PK standpoint. Preliminary analysis suggests that higher exposures are associated with greater neutrophil decrease in the circulation, leading to better efficacy, as measured by IHS4 and HiSCR50 / HiSCR75. Because HS is an aggressive disease and due to the relatively short half-life of the drug and likely low bioavailability at the site of action [Li et al., 2021, 48:743] when administered SC, it is deemed appropriate to dose subjects more frequently. Hence, a Q2W regimen will be tested in Study CSL324_2001.

[0897] Study CSL324_2001 will test the efficacy and safety of a single SC dose, equivalent to 0.6 mg / kg IV in subjects with HS in Treatment Period A. Because the median body weight of subjects with HS in Study CSL324_1002 is about 90 kg, coupled with the average SC bioavailability of 73% based on NCA-derived exposure data (AUCo-inf) following cross-study analysis and a shorter half-life after SC administration, it appears that a dose of 50 mg Q2W will yield similar exposure to a 0.6 mg / kg Q3W IV dose, where safety and clinical responses were previously demonstrated. Similarly, a fixed SC dose of 25 mg Q2W is estimated to yield an exposure similar to a 0.3 mg / kg Q3W dose when administered IV.

[0898] The relationship between anumigilimab serum concentrations and circulating neutrophils following single and repeat dose administration in Studies CSL324_1001, CSL324_1002, and CSL324_1003 was adequately described by a semi-mechanistic model of the neutrophil lifecycle [Mangas-Sanjuan et al., 2015, 354:55]. Simulations from this model predicted that < 15% of subjects with HS in the current study may experience severe neutropenia when dosed SC with 50 mg anumigilimab Q2W for 16 weeks, and < 18% of subjects with HS in the current study may experience severe neutropenia when dosed SC with 50 mg anumigilimab Q2W for 32 weeks. However, these predictions assumed a continual time-dependent decline in ANC values and carry a degree of uncertainty due to its extrapolation of dosing duration beyond the clinical experience of Study CSL324_1002 (5 doses) and the small sample size. Additional risk mitigation measures as described in the section entitled “Risk Assessment” above, are implemented in this study to further minimize the incidence of moderate or severe neutropenia.

[0899] Model-based simulation was performed to explore total exposures (area under the concentration-time curve [AUC]) at different body weight cutoffs using fixed exponents for body weight on clearance and volume of distribution. A flat dose of 50 (or 25) mg SC provides comparable exposures, between 80% and 125% of the reference AUC (based on a body weight of 90 kg) across a wide range of body weights.

[0900] In summary, a dose of 50 mg administered to subjects SC Q2W provides a reasonable dose regimen based on both observed and simulated results of anumigilimab exposure and neutrophil profiles from 3 phase 1 studies. Start of Study Definition

[0901] The study start date is the date on which the clinical study will be open for recruitment of subjects. The first act of recruitment is the time point when the first site is activated and will be considered the study start date.

[0902] End of Study Definition

[0903] The end of the study is defined as the date of the last scheduled procedure shown in the section entitled “Schedule of Activities” above for the last subject in the study globally. A subject is considered to have completed the study if they completed all study periods, including the last scheduled procedure shown in the section entitled “Schedule of Activities” above.

[0904] Study sites will be closed upon study completion. A study site is considered closed when all required documents and study supplies have been collected and a study closeout visit has been performed.

[0905] STUDY POPULATION

[0906] Subjects must meet all inclusion criteria and none of the exclusion criteria to be eligible for enrolment into this study. Prospective approval of protocol deviations from eligibility criteria, also known as protocol waivers or exemptions, is not permitted.

[0907] Inclusion Criteria

[0908] To be enrolled into the study, subjects must meet all of the following inclusion criteria:

[0909] 1. Capable of providing written informed consent and willing and able to adhere to all protocol requirements.

[0910] 2. Male or female between > 18 and < 75 years of age at the time of providing written informed consent.

[0911] 3. Confirmed clinical diagnosis of HS at least 6 months before Screening and moderate to severe HS as per IHS4 guidelines (ie, IHS4 > 4) at the Screening Visit with at least 1 draining tunnel.

[0912] 4. Stable disease for at least 2 months before Screening, as determined by the investigator through review of medical history and subject recall.

[0913] 5. Inadequate response to at least a 3-month (90-day) trial of oral antibiotics for the treatment of HS (or demonstrated intolerance to, or had a contraindication to, oral antibiotics for treatment of their HS) as determined by the investigator through review of medical history and subject recall.

[0914] 6. Inadequate clinical response to at least 12 weeks of treatment with anti-TNF blocking agents as assessed by the investigator.

[0915] 7. Must agree to use antiseptic wash daily on HS-affected areas (eg, 4% chlorhexidine) during the study.

[0916] Exclusion Criteria

[0917] Subjects must not be enrolled into the study if they meet any of the following exclusion criteria:

[0918] 1. Treatment with any medications and therapies not permitted during the study.

[0919] 2. History of myeloproliferative disease.

[0920] 3. Malignancy within 5 years at Screening, with the exception of non-malignant skin cancer, carcinoma in situ, or prostate cancer not requiring treatment.

[0921] 4. Current or a recent clinically significant history of uncontrolled renal, hepatic (including currently active hepatitis B virus and / or hepatitis C virus), hematologic, endocrine, neurologic, pulmonary, psychiatric, or cardiac disease, assessed as potentially having an effect on subject safety or study outcomes as determined by the investigator and / or sponsor.

[0922] 5. Primary or secondary immunodeficiencies, including human immunodeficiency virus infection.

[0923] 6. Clinical signs of active infection and / or fever > 38°C during the 7 days before Day 1; OR history of chronic, serious, or recurrent infections; OR, in the 2 months before Screening, has been hospitalized or received IV antibiotic therapy for an infection. Study entry may be deferred and subject rescreened, if required, at the investigator’s and / or sponsor’s discretion.

[0924] Note: HS will not be considered an infection for this criterion. Antibiotic use to treat HS is described in Table 11.

[0925] 7. ANC < 2.0 x 109cells / L at the Screening Visit.

[0926] 8. Subjects with > 20 draining tunnels.

[0927] 9. Any other skin conditions that, in the opinion of the investigator, interfere with the assessment of HS lesions and disease severity.

[0928] 10. Known history of hypersensitivity reactions or suspected hypersensitivity to the active substance or to any of the excipients of anumigilimab or other monoclonal antibodies (mAbs). 11. Previously administered anumigilimab in another study.

[0929] 12. Female subject of childbearing potential or fertile male subject either not using or not willing to use an acceptable method of contraception to avoid pregnancy during the study until the End of Study Visit.

[0930] All female subjects are assumed to be of childbearing potential except: a. Subjects aged > 60 years. b. Subjects aged 45 to 60 years (inclusive) with amenorrhea for > 1 year with documented evidence of follicle-stimulating hormone (FSH) level > 30 IU / L. If the FSH value is not available before randomization, a urine pregnancy test is required. c. Subjects who are surgically sterile for at least 3 months before providing informed consent.

[0931] All male subjects are assumed fertile except subjects who are surgically sterile for at least 3 months before providing informed consent.

[0932] 13. Female subject who is pregnant, breastfeeding, or not willing to cease breastfeeding.

[0933] 14. Any issue including clinically significant abnormalities in physical examination findings, electrocardiogram (ECG) readings, and safety laboratory test results that, in the opinion of the investigator or sponsor, would render the subject unsuitable for participation in the study.

[0934] 15. Involved in the planning and / or conduct of the study (applies to CSE Behring [CSE] staff, staff at the study site, and third-party vendors).

[0935] 16. Participated in another interventional clinical study within 30 days (or 5 halflives, whichever is longer) before the first administration of anumigilimab.

[0936] Lifestyle Considerations

[0937] Subjects are expected to continue their normal lifestyle during the study and to avoid events and activities that would prevent them from following the protocol-specified study site visits in person.

[0938] Women of childbearing potential (WOCBP) (as defined in the section entitled “Definitions” below) and all male subjects who have partners considered to be WOCBP must use effective methods of contraception, from the first dose of IP until 3 months after the subject’s final study visit.

[0939] Screen Failure

[0940] A screen failure is defined as a subject who consents to participate in the clinical study but does not meet the eligibility criteria. If a subject is not eligible for the study (ie, screen failure), minimal information should be recorded in the electronic case report form (eCRF; i.e., demography, screen failure details, eligibility criteria, and any serious adverse events [SAEs]) to meet the Consolidated Standards of Reporting Trials (CONSORT) publishing requirements and to respond to queries from regulatory authorities.

[0941] Subjects who do not meet the criteria for participation in this study (screen failure) may be rescreened. Rescreening is permitted as follows:

[0942] • A subject may be rescreened after approval from the sponsor.

[0943] • A subject may be rescreened once.

[0944] • The retesting of abnormal screening tests, which are exclusionary and considered potentially erroneous, is allowed at an unscheduled visit during the Screening Period (to reassess eligibility). This may not be considered rescreening.

[0945] If a subject is rescreened, all screening assessments must be repeated.

[0946] If a subject is rescreened, a new informed consent form (ICF) must be signed (Section entitled “Informed Consent Process” below).

[0947] Rescreened subjects should retain their originally assigned subject number through every screening / re screening event.

[0948] STUDY INTERVENTIONS AND CONCOMITANT THERAPY

[0949] Investigational Products

[0950] The IP should only be dispensed or administered to subjects currently enrolled in the study.

[0951] Anumigilimab will be supplied as a sterile liquid formulation at a concentration of 50 mg / mL (in 2-mL vials; Table 10), formulated in a buffered solution containing stabilizing excipients. The liquid formulation consists of 50 mg / ml CSL324, 20 mM L- histidine buffer, 100 mM L-Arginine-HCl, 100 mM L-Proline, and 0.03% w / w PS80, at pH 5.7. Each excipient is of a grade that is suitable for pharmaceutical manufacture and has been used in currently marketed formulations of mAbs. Anumigilimab will be manufactured by the sponsor in accordance with International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) Good Manufacturing Practice guidelines and local regulatory requirements.

[0952] Anumigilimab has not yet been approved for the treatment of HS in any country. No auxiliary medicinal products as defined by Article 2(8) of Regulation (European Union [EU]) 536 / 2014 will be used in this study. Table 10. Investigational Product - Pharmaceutical Properties, Formulation, and

[0953] Administration Characteristics.

[0954] INN = international nonproprietary name; N / A = not applicable. Anumigilimab will be administered by the investigator at the study site on Day 1,

[0955] Day 15, Day 29, Day 43, Day 57, Day 71, Day 85, Day 99, and Day 112 in Treatment Period A and on Day 127, Day 141, Day 155, Day 169, Day 183, Day 197, and Day 211 in Treatment Period B. Detailed information on the preparation and administration of anumigilimab is provided in the Site Investigational Medicinal Product Manual. Anumigilimab will be supplied to the study sites by the sponsor or delegate.

[0956] Selection of Doses in the Study

[0957] The dose to be tested in this study is 50 mg of anumigilimab or placebo Q2W, administered SC for a total of 16 weeks (blinded Treatment Period A). Following that, subjects in the 50 mg anumigilimab or placebo arms will either receive a 25 mg SC or 50 mg SC anumigilimab Q2W for an additional 16 weeks in the blinded Treatment Period B (see Figure 6). Dose selection for the study is described in the section entitled “Dose Rationale” above.

[0958] Selection and Timing of Dose for Each Subject

[0959] Each subject in Treatment Period A will receive either a fixed dose of 50 mg anumigilimab or placebo Q2W for a total of 16 weeks. Completers from each arm in Treatment Period A will be (re-)randomized in Treatment Period B in a ratio of 1:1 to anumigilimab 50 mg SC Q2W and 25 mg SC Q2W arms. Each subject in Treatment Period B will either receive 25 mg SC or 50 mg SC anumigilimab Q2W for an additional 16 weeks.

[0960] Anumigilimab will be administered at the study site at the time points specified in the section entitled “Schedule of Activities” above. Detailed information on the preparation and administration of anumigilimab is provided in the Site Investigational Medicinal Product Manual.

[0961] Dose Modification

[0962] Dose modification is not allowed in this study.

[0963] Prior and Concomitant Therapy

[0964] Any medication or vaccine (including over-the-counter or prescription medicines, recreational drugs, vitamins, and / or herbal supplements) or other specific categories of interest that the subject is receiving up to 30 days before Screening or receives during the subject’s study participation must be recorded along with the following:

[0965] • Indication.

[0966] • Dates of administration including start and end dates, if applicable.

[0967] • Dosage information including dose and frequency.

[0968] Additionally, any anti-TNF therapy, biologies, or any disease-modifying agents given for the treatment of HS, regardless of how long ago given, should also be recorded. The reason for discontinuation should also be recorded, along with perceived benefit and loss of response as assessed by the investigator from the subject’s medical records or medical history based on the subject’s recollection.

[0969] The sponsor’s medical monitor should be contacted if there are any questions regarding prior or concomitant therapy.

[0970] Prohibited and permitted prior or concomitant therapies are summarised in Table 11 and Table 12 below, respectively.

[0971] Table 11. Permitted Prior and Concomitant Therapies.

[0972] Table 12. Prohibited Prior and Concomitant Therapies.

[0973] BCG = Bacillus Calmette Guerin; TNF = tumor necrosis factor.

[0974] SUBJECT WITHDRAWAL AND DISCONTINUATION OF INVESTIGATIONAL PRODUCT

[0975] Discontinuation of Investigational Product

[0976] Subjects may discontinue IP at any time at their own request, or at the discretion of the investigator or the sponsor for safety, behavioral, or administrative reasons. Additionally, further treatment with anumigilimab will be discontinued if any of the following criteria are met.

[0977] 1. An SAE related to the IP.

[0978] 2. Hypersensitivity reaction to anumigilimab or excipients. 3. Severe neutropenia (ANC < 0.5 x 109cells / L) that is confirmed on repeat measurement approximately 24 hours later.

[0979] 4. Moderate neutropenia (ANC < 1.0 x 109cells / L to > 0.5 x io9cells / L) that is confirmed by repeat measurement approximately 24 hours later and is sustained until the next planned administration of IP. Administration of anumigilimab can be resumed if ANC is > 1.0 x io9cells / L at the next originally planned dosing day. ANC threshold for resumption of IP administration can be revised, if necessary, by IDMC after review of safety data.

[0980] 5. Development of febrile neutropenia (moderate neutropenia [ANC < 1 x 109cells / L] and a single tympanic temperature [preferred] of > 38.3 °C or oral temperature

[0981] > 38.0°C sustained for > 1 hour). Administration of anumigilimab can be resumed if ANC is > 1.0 x 109cells / L and clinically significant infections have been ruled out or subject has recovered at the next originally planned dosing day.

[0982] 6. In the investigator’s or sponsor’s opinion, continued administration of IP could be detrimental to the subject’s wellbeing.

[0983] Subjects who permanently discontinue IP will undergo the assessments scheduled for the Day 225 (End of Treatment) Visit and will subsequently attend the Follow-up Visits (Day 252 and Day 280, 4 and 8 weeks after the last IP administration, respectively) as detailed in the section entitled “Schedule of Activities” above. Subjects discontinuing IP who decline further study procedures / visit participation will be withdrawn from the study (Section entitled “Subject Withdrawal From the Study” below).

[0984] Temporary Interruption

[0985] ANC will be checked within 2 days prior to each dosing of IP. If ANC is < 1 x 109cells / L at this time point, dosing will be interrupted. Administration of anumigilimab can be resumed if ANC is > 1.0 x 109cells / L at the next originally planned dosing day. ANC threshold for resumption of IP administration can be revised, if necessary, by the IDMC after review of safety data. Dosing should otherwise not be interrupted in the study unless discussed with the sponsor’s medical monitor.

[0986] Halting or Stopping Criteria for Investigational Product

[0987] If any of the following events occur during the study, recruitment will be halted and dosing for enrolled subjects suspended.

[0988] • An SAE considered related to the IP either by the investigator or sponsor that results in death. • One or more subject(s) develops any AE (including laboratory abnormality of clinical significance) considered related to the IP and deemed to pose an unacceptable risk to other subjects in the study as assessed by the sponsor.

[0989] If the safety concerns warrant stopping the study permanently, the IDMC will advise the CSL Global Safety Governance Committees, which in turn will conduct a risk assessment. The CSL Global Safety Governance Committees, in agreement with the IDMC, will recommend that the study be stopped if it is concluded that continued dosing poses an unacceptable risk to subjects and no further risk mitigation steps can be applied. Regulatory authorities will be notified of a final study stop.

[0990] Rechallenge

[0991] Restart of IP after temporary interruption due to ANC < 1 x 109cells / L will be performed if the subject’s ANC improves to > 1 x 109cells / L at the next dosing visit as described in the section entitled “Temporary Interruption” above.

[0992] STUDY ASSESSMENTS AND PROCEDURES

[0993] • Study assessments and procedures and their timing are summarized in the section entitled “Schedule of Activities” above. Protocol waivers or exemptions are not allowed.

[0994] • The investigator should discuss any safety concerns with the sponsor’s medical monitor or delegate immediately upon occurrence or when the investigator becomes aware of the safety concern to determine if the subject should continue or discontinue IP.

[0995] • Adherence to the study design requirements, including those specified in section entitled “Schedule of Activities” above, is essential and required for study conduct.

[0996] • All screening evaluations must be completed and reviewed to confirm that potential subjects meet all eligibility criteria. The investigator will maintain a screening log to record details of all subjects screened and to confirm eligibility or record reasons for screen failure, as applicable.

[0997] • Procedures conducted as part of the subject’s routine standard of care (eg, blood count) and obtained before signing of the ICF may be used for screening or baseline purposes provided the procedures met the protocol- specified criteria and were performed within the time frame defined in the section entitled “Schedule of Activities” above.

[0998] • ANC results will not be shared with the blinded examiner until the study has been unblinded.

[0999] • Repeat or unscheduled samples may be taken for safety reasons or for technical issues with the samples. Refer to the section entitled “Study Assessment and Procedure Considerations” below for a description of the assessments and visits which may be performed in a remote / off-site setting.

[1000] Screening / Baseline Assessments

[1001] Demographics

[1002] The following demographic information will be collected from subjects at the Screening Visit:

[1003] • Year of birth.

[1004] • Age.

[1005] • Sex.

[1006] • Ethnicity (where permitted).

[1007] • Smoking status.

[1008] Medical History

[1009] The following information on the subject’s medical history will be collected at the Screening Visit:

[1010] • Relevant medical history.

[1011] • Diagnosis and disease status.

[1012] • Contraceptive method used (if relevant).

[1013] Physical Examination

[1014] A physical examination will be performed per the study site’s standard procedure.

[1015] Dermatological Examination for HS Lesions by the Blinded Examiner

[1016] A blinded examiner will perform a baseline dermatological examination for HS lesions as detailed in the section entitled “Blinded Examiner” above.

[1017] Vital Signs, Weight, and Height

[1018] Vital signs and weight will be collected as indicated in Table 13. Height will only be collected at Screening.

[1019] Pregnancy Test

[1020] A serum pregnancy test will be performed as indicated in Table 13. Clinical Laboratory Assessments

[1021] Laboratory tests for urinalysis, hematology, biochemistry, and viral serology will be performed as indicated in Table 13. A test for serum FSH may be performed at Screening to confirm postmenopausal status is female subjects aged 45 to 60 years (inclusive) with amenorrhea for > 1 year.

[1022] Collection of Prior and Concomitant Therapies and Adverse Events

[1023] Collection of prior and concomitant therapies will be performed as indicated in the section entitled “Prior and Concomitant Therapy” above. Collection of AEs will be performed as indicated in the section entitled “Adverse Events and Serious Adverse Events: Definitions and Procedures for Recording, Evaluating, Follow-up, and Reporting” below.

[1024] Efficacv Assessments

[1025] 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.

[1026] Efficacy assessments will be performed at the time points specified in the section entitled “Schedule of Activities” above.

[1027] International Hidradenitis Suppurativa Severity Score System

[1028] The IHS4 is a dynamic scoring system developed and clinically validated to assess disease severity of HS [Zouboulis et al., 2017, 177: 1401]. 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],

[1029] 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],

[1030] Hidradenitis Suppurative Clinical Response

[1031] HiSCR, also referred to herein as HiSCR50, was developed and validated in 2014 [Kimball et al., 2014, 171: 1434], The HiSCR is a valid, responsive, and meaningful clinical endpoint of the inflammatory manifestations of HS that can be adapted to clinical research and daily practice. 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],

[1032] 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.

[1033] Patient-reported Outcomes

[1034] The patient-reported outcomes that will be assessed during the study are the DLQI, pain NRS score, and HiSQOL. The assessments will be done at the sites using an electronic clinical outcomes assessment solution. Patient-reported outcomes should be completed ahead of other study procedures, except for ANC determination, at each study visit. Dermatology Life Quality Index

[1035] The DLQI is a validated questionnaire consisting of 10 questions that assess the effect of skin conditions on the quality of life of an affected patient [Finlay and Khan, 1994, 19:210]. Use of the DLQI has been described in more than 1000 publications, including many multinational studies, for more than 40 types of skin conditions and in more than 80 countries, and is available in more than 90 languages. Each question is scored from 0 (not at all) to 3 (very much), with a recall period of 1 week, and scores are tallied to produce a total score. Total scores are graded as follows: 0 to 1 indicates no effect, 2 to 5 indicates a small effect, 6 to 10 indicates a moderate effect, 11 to 20 indicates a very large effect, and 21 to 30 (maximum score) indicates an extremely large effect on the patient’s life [Hongbo et al., 2005, 125:659]. Subjects are to complete DLQI at the study site, as indicated in the section entitled “Schedule of Activities” above.

[1036] Pain Numerical Rating Scale

[1037] The pain NRS is a unidimensional measure of pain intensity in adults, including those with chronic pain associated with skin conditions [Kimball et al., 2016b, 375:422]. The NRS is a segmented numeric version of the visual analog scale in which a respondent selects a whole number (from 0 to 10) that best reflects the intensity of their pain over the last 24 hours [Rodriguez, 2001, 2:38]. The common format is a horizontal bar or line that is anchored by terms describing pain severity extremes [Hawker et al., 2011, 63:S240]. The pain NRS score will be collected at the study site, as indicated in the section entitled “Schedule of Activities” above. This assessment will document a subject’s global assessment of skin pain over the previous 24 hours and the previous 7 days, capturing both pain at its worst and pain on average for the respective time periods.

[1038] Hidradenitis Suppurativa Quality of Life

[1039] The HiSQOL was developed to measure changes in health -related quality of life in patients with HS [Kirby et al., 2020]. The 17 -item HiSQOL included 4 symptom items, 8 activity adaptation items, and 5 psychosocial items. Each question is scored from 4 (extremely) to 0 (not at all), with a recall period of 1 week. The item scores are summed to create a total score ranging from 0 to 68, with higher scores indicating more severe impact on health -related quality of life. The subscale scores range from 0 to 16 for symptoms, 0 to 20 for psychosocial, and 0 to 32 for activities-adaptations [Kirby et al., 2020, 183:340]. Subjects are to complete the HiSQOL at the study site, as indicated in the section entitled “Schedule of Activities” above. Safety Assessments

[1040] Safety assessments will be performed at the time points specified in the section entitled “Schedule of Activities” above. Table 13. Safety Assessments.

[1041] AE = adverse event; ANC = absolute neutrophil count; ECG = electrocardiogram; FDA = Food and Drug Administration; HIV = human immunodeficiency virus; hs-CRP = high-sensitivity C-reactive protein; POC = point of care; QTcB = QT interval corrected with Bazett formula; QTcF = QT interval corrected with Fridericia formula; SAE = serious adverse event.aAnalysis will be conducted at a central laboratory. Additional details are provided in the Laboratory Manual.

[1042] Vital Signs

[1043] Vital signs will be collected as indicated in Table 13.

[1044] Electrocardiograms

[1045] Electrocardiogram parameters specified in Table 13 will be measured. Central reading of ECGs will be performed. Further details are provided in the respective manual. Clinical Safety Laboratory Assessments

[1046] • The investigator must review the laboratory report, document this review, and record any clinically significant changes occurring during the study as an AE. The laboratory reports must be filed with the source documents. • Abnormal laboratory findings associated with the underlying disease are not considered clinically significant unless judged by the investigator to be more severe than expected for the subject’s condition.

[1047] • All laboratory tests with values considered clinically significantly abnormal during participation in the study or within 8 weeks after th...

Claims

CLAIMS1. A method of treating a neutrophilic skin condition in a human subject suffering from the neutrophilic skin condition, the method comprising administering subcutaneously to the subject a fixed dose of 25 mg or 50 mg of an antibody that inhibits granulocyte colony stimulating factor (G-CSF) signaling every 13-15 days.

2. The method of claim 1, wherein:(i) the antibody is administered every 13-15 days during a first dosing period; and / or(ii) the antibody is administered every 13-15 days during a second dosing period.

3. A method of treating a neutrophilic skin condition in a human subject suffering from the neutrophilic skin condition, the method comprising:(i) administering subcutaneously to the subject a fixed dose of 25 mg or 50 mg of an antibody that inhibits granulocyte colony stimulating factor (G-CSF) signaling during a first dosing period; and(ii) subsequently administering subcutaneously to the subject a fixed dose of 25 mg or 50 mg of the antibody that inhibits G-CSF during a second dosing period, wherein the antibody that inhibits G-CSF is administered every 13-15 days during the first dosing period and / or the second dosing period.

4. The method of claim 2 or claim 3, wherein the first dosing period and / or the second dosing period is at least 16 weeks.

5. The method of any one of claims 1 to 4, wherein the subject has previously received treatment with an anti-tumour necrosis factor (TNF) therapy.

6. The method of claim 5, wherein the subject has had an inadequate clinical response to at least 12 weeks of treatment with the anti-TNF therapy.

7. The method of claims 5 or 6, wherein the anti-TNF therapy is an anti-TNFa antibody.

8. The method of claim 7, wherein the anti-TNFa antibody is adalimumab.

9. The method of any one of claims 1 to 8, 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 neutrophilic skin condition.

10. The method of any one of claims 1 to 9, wherein administration of the antibody does not cause sustained grade 3 or grade 4 neutropenia in the subject for greater than seven consecutive days.

11. The method of any one of claims 1 to 10, wherein administration of the antibody:(i) does not induce grade 4 neutropenia; or(ii) does not induce neutropenia or the antibody induces grade 2 or grade 3 neutropenia for two consecutive days or less; or(iii) does not induce neutropenia for more than 2 consecutive days or more than 1 day.

12. The method of claim 10 or claim 11, wherein the neutropenia is not associated with a fever.

13. The method of any one of claims 1 to 12, wherein the subject has an absolute neutrophil count (ANC) of > 2.0 x 109 / L prior to receiving treatment with the antibody that inhibits G-CSF signaling.

14. The method of any one of claims 1 to 13, wherein the subject does not have one or more of the following prior to receiving treatment with the antibody:(i) a history of myeloproliferative disease;(ii) malignancy within 5 years of receiving treatment with the antibody;(iii) a clinically significant history of uncontrolled renal, hepatic (including currently active hepatitis B virus and / or hepatitis C virus), hematologic, endocrine, neurologic, pulmonary, psychiatric and / or cardiac disease;(iv) a primary or secondary immunodeficiency;(v) clinical signs of active infection and / or fever > 38°C during the 7 days before receiving treatment with the antibody;(vi) a history of chronic, serious, or recurrent infections;(vii) has been hospitalized or received intravenous antibiotic therapy for an infection within 2 months of receiving treatment with the antibody; and(viii) has > 20 draining tunnels.

15. The method of any one of claims 1 to 14, wherein the neutrophilic skin condition is a neutrophilic dermatosis or a neutrophilic skin lesion.

16. The method of claim 15, wherein the neutrophilic dermatosis is selected from the group consisting of hidradenitis suppurativa (HS); palmoplantar pustulosis (PPP); amicrobial pustulosis of the folds (APF); psoriasis; plaque psoriasis; CARD14-mediated pustular psoriasis (CAMPS); cryopyrin associated periodic syndromes (CAPS); deficiency of interleukin- 1 receptor (DIRA); deficiency of interleukin- 36 receptor antagonist (DIRTA); pyogenic arthritis; pyoderma gangrenosum and acne (PAPA); pyoderma gangrenosum, acne, and hidradenitis suppurativa (PASH); pyoderma gangrenosum (PG); skin lesions of Behcet’s disease; Still’s disease; Sweet syndrome; subcorneal pustulosis (Sneddon-Wilkinson); pustular psoriasis; palmoplantar pustulosis; acute generalized exanthematic pustulosis; infantile acropustulosis; synovitis, acne, pustulosis; hyperostosis and osteitis (SAPHO) syndrome; bowel-associated dermatosis-arthritis syndrome (BAD AS); neutrophilic dermatosis of the dorsal hands; neutrophilic eccrine hidradenitis; erythema elevatum diutinum; and Pyoderma gangrenosum.

17. The method of claims 15 or 16, wherein the neutrophilic dermatosis is hidradenitis suppurativa (HS).

18. The method of claim 17, wherein the subject is suffering from moderate to severe HS and / or has an International Hidradenitis Suppurativa Severity Score System (IHS4) score of > 4.

19. A method of treating hidradenitis suppurativa (HS) in a subject, wherein the subject:(i) is suffering from moderate to severe HS and / or has an International Hidradenitis Suppurativa Severity Score System (IHS4) score of > 4; and(ii) has had an inadequate response to an anti-tumour necrosis factor (TNF) therapy, the method comprising:(i) administering subcutaneously to the subject a fixed dose of 25 mg or 50 mg of an antibody that inhibits granulocyte colony stimulating factor (G-CSF) signaling during a first dosing period; and(ii) subsequently administering subcutaneously to the subject a fixed dose of 25 mg or 50 mg of the antibody that inhibits G-CSF during a second dosing period, wherein the antibody that inhibits G-CSF is administered every 13-15 days during the first dosing period and / or the second dosing period.

20. A method of treating hidradenitis suppurativa (HS) in a subject, wherein the subject:(i) is suffering from moderate to severe HS and / or has an International Hidradenitis Suppurativa Severity Score System (IHS4) score of > 4; and(ii) has had an inadequate response to an anti-tumour necrosis factor (TNF) therapy, the method comprising:(i) administering subcutaneously to the subject a fixed dose of 25 mg or 50 mg of an antibody that inhibits granulocyte colony stimulating factor (G-CSF) signaling during a first dosing period; and(ii) subsequently administering subcutaneously to the subject a fixed dose of 25 mg or 50 mg of the antibody that inhibits G-CSF during a second dosing period, wherein the antibody that inhibits G-CSF is administered every 13-15 days during the first dosing period and the second dosing period, and wherein the first dosing period is at least 16 weeks.

21. The method of any one of claims 17 to 19, wherein the subject has stable HS for at least 2 months and / or at least 1 draining tunnel prior to receiving treatment.

22. The method of any one of claims 17 to 21, wherein the antibody that inhibits G-CSF is administered in an amount sufficient to reduce or prevent an increase in the subject’s IHS4 score.

23. The method of claim 22, wherein following 16 weeks of treatment the subject’s IHS4 score is(i) reduced by at least 2, at least 4, at least 6, at least 8 or at least 10; and / or(ii) reduced by at least 55%.

24. A method of treating hidradenitis suppurativa (HS) in a subject, wherein the subject:(i) is suffering from moderate to severe HS and / or has an International Hidradenitis Suppurativa Severity Score System (IHS4) score of > 4; and(ii) has had an inadequate response to an anti-tumour necrosis factor (TNF) therapy, the method comprising administering subcutaneously to the subject a fixed dose of 25 mg or 50 mg of an antibody that inhibits granulocyte colony stimulating factor (G-CSF) signaling every 13-15 days, and wherein an absolute reduction in the IHS4 score is achieved following a first dosing period of at least 16 weeks.

25. A method of reducing or preventing an increase in an International Hidradenitis Suppurativa Severity Score System (IHS4) score in a subject, wherein the subject is suffering from moderate to severe HS and / or has an IHS4 score of > 4, the method comprising administering subcutaneously to the subject a fixed dose of 25 mg or 50 mg of an antibody that inhibits granulocyte colony stimulating factor (G-CSF) signaling every 13-15 days, and wherein an absolute reduction in the IHS4 score is achieved following a first dosing period of at least 16 weeks.

26. The method of claim 25, the method further comprising subsequently administering subcutaneously to the subject a fixed dose of 25 mg or 50 mg of the antibody that inhibits G-CSF every 13-15 days during a second dosing period, wherein the reduction in the IHS4 score is maintained during the second dosing period.

27. A method for maintaining a reduction of an International Hidradenitis Suppurativa Severity Score System (IHS4) score in a subject suffering from Hidradenitis Suppurativa (HS) and who has achieved an absolute reduction in IHS4 score from baseline during a first dosing period, the method comprising administering subcutaneously to the subject a fixed dose of 25 mg or 50 mg of the antibody that inhibits G-CSF every 13-15 days during a second dosing period.

28. The method of any one of claims 22 to 27, wherein the IHS4 score is reduced by > 55% (IHS4-55) at the end of a dosing period of at least 16 weeks.

29. The method of any one of claims 22 to 28, wherein one or more of the following is achieved in the subject at the end of a first dosing period of at least 16 weeks:(i) a hidradenitis suppurativa clinical response (HiSCR) 50 or HiSCR 75 or modified HiSCR response;(ii) a reduction in a Dermatology Life Quality Index (DLQI) score from baseline;(iii) a reduction in the number of inflammatory lesions (AN counts) from baseline; and(iv) a reduction in Patient’s Global Assessment of Skin Pain including: a) pain at its worst and pain on average over the previous 24 hours and / or b) pain at its worst and pain on average over previous 7 days.

30. A method of treating hidradenitis suppurativa (HS) in a subject, wherein the subject:(i) is suffering from moderate to severe HS and / or has an International Hidradenitis Suppurativa Severity Score System (IHS4) score of > 4; and(ii) has had an inadequate response to an anti-tumour necrosis factor (TNF) therapy, the method comprising administering subcutaneously to the subject a fixed dose of 25 mg or 50 mg of an antibody that inhibits granulocyte colony stimulating factor (G-CSF) signaling every 13-15 days, and wherein following a first dosing period of at least 16 weeks one or more of the following is achieved:(i) at least a 55% reduction in the IHS4 score;(ii) a hidradenitis suppurativa clinical response (HiSCR) 50 or HiSCR 75 or modified HiSCR response;(iii) a reduction in a Dermatology Life Quality Index (DLQI) score from baseline;(iv) a reduction in the number of inflammatory lesions (AN counts) from baseline; and(v) a reduction in Patient’ s Global Assessment of Skin Pain including: a) pain at its worst and pain on average from baseline; b) pain at its worst and pain on average over the previous 24 hours and / or c) pain at its worst and pain on average over previous 7 days.

31. The method of claim 30, wherein the reduction in Patient’s Global Assessment of Skin Pain is at least a 30% reduction in numeric rating scale (NRS) and at least 1-unit reduction from baseline in worst skin pain.

32. The method of any one of claims 1 to 31, wherein the antibody binds to G-CSFR and inhibits G-CSF signalling.

33. The method of any one of claims 1 to 32, wherein the antibody binds to or specifically binds to granulocyte-colony stimulating factor receptor (G-CSFR) andcompetitively 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.

34. The method of any one of claims 1 to 33, wherein the antibody 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.

35. The method of any one of claims 1 to 34, wherein the antibody comprises:(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.

36. The method of any one of claims 1 to 35, wherein the antibody comprises:(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 SEQ ID NO: 28.

37. The method of any one of claims 1 to 36, wherein the antibody comprises:(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: 18 and a light chain comprising a sequence set forth in SEQ ID NO: 15.

38. The method of any one of claims 1 to 37, wherein the antibody is administered in a pharmaceutical formulation comprising the antibody at a concentration of 50 mg / mL, wherein the formulation has a pH of 5.5 to 5.9 and comprises 12 mM to 30 mM histidine buffer, 0.02 % to 0.04 % (w / v) polysorbate 80, 95 mM to 105 mM proline, and 90 mM to 110 mM arginine.

39. The method of claim 38, wherein the formulation comprises:(i) 20 mM histidine buffer;(ii) 0.03% (w / v) polysorbate 80;(iii) 100 mM arginine; and / or(iv) 100 mM proline.

40. The method of any one of claims 1 to 39, wherein the antibody is administered every 14 days.

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