Bispecific antibodies for use in the treatment of hidradenitis suppurativa - Patent Application 20070233334

Abstract

The present invention relates to bivalent, bispecific monoclonal antibodies (bbmAbs) or variants thereof for use in treating hidradenitis suppurativa in a subject or for use in alleviating the symptoms thereof.

Description

[Technical field]

[0001] The present invention relates to bivalent, bispecific monoclonal antibodies (bbmAbs) or variants thereof for use in treating patients suffering from hidradenitis suppurativa. The present disclosure also relates to methods and therapeutic regimens for treating hidradenitis suppurativa by using bispecific antibodies that simultaneously target both IL-1β and IL-18. [Background technology]

[0002] Hidradenitis suppurativa (HS), also called "acne inversa" or "maladie de Verneuilh", is a chronic, recurrent and debilitating inflammatory skin condition that typically presents as deep, inflammatory and painful lesions in areas of the body that contain apocrine glands. The most commonly affected areas are the axillae, groin and anogenital region (Jemec 2012; Fimmel and Zouboulis 2010).

[0003] HS is currently considered to be an inflammatory disease of the pilosebaceous follicle based on an immune system imbalance occurring in genetically predisposed individuals (Kelly et al., 2014). While thought to be primarily caused by follicular obstruction, HS is an inflammatory skin disease characterized by large numbers of neutrophils and macrophages in inflammatory foci (Lima et al., 2016; Shah et al., 2017). While the pathophysiology of HS remains largely unknown, tumor necrosis factor alpha (TNFα) blockade has been described to be beneficial in larger studies (Kimball et al., 2016). Evidence of efficacy of anti-IL1 treatment (Tzanetakou et al. 2016) and blockade of IL-17A (Thorlacius et al. 2017; Schuch et al. 2018; Giuseppe et al. 2018; Jorgensen et al. 2018) or anti-IL-23 treatment (Sharon et al. 2012; Blok et al. 2016) has also been found in smaller studies and / or case reports. More recently, research approaches using the oral PDE4 inhibitor apremilast (Weber et al. 2017) or anti-complement 5a compounds (Kanni et al. 2018) have been described.

[0004] The disease begins after puberty and affects women more frequently than men (3:1). Risk factors include obesity and smoking. Epidemiological prevalence estimates vary widely (0.03-4.3%; Jemec 2012; Jemec and Kimball 2015), and although geographic differences exist, a prevalence of approximately 0.1-1% is accepted by the scientific community (Garg et al. 2018).

[0005] Although the clinical manifestations of HS are heterogeneous, the disease tends to manifest with chronic, recurrent, deep, painful inflammatory skin lesions, mostly with inflammatory nodules and abscesses that may lead to drainage and suppuration. The inflammatory lesions may be complicated by sinus tract and fistula formation during disease progression, leading to hypertrophic scarring that may impact on functional use.

[0006] HS often has devastating psychosocial impacts, with pain, foul-smelling discharge from wounds and scarring. HS is a severely debilitating disease with a significant negative impact on quality of life (QoL), with multiple studies confirming that the impact is greater than that seen with other skin diseases (Deckers and Kimball 2016). HS patients also often suffer from depression, social isolation, which can impair sexual health and make it difficult for patients to perform their job functions (Esmann and Jemec 2011, Fimmel and Zouboulis 2010, Janse et al. 2017).

[0007] HS is difficult to treat. Official European treatment guidelines were only drawn up in 2015, which suggest that patients should be offered adjuvant medical and surgical treatment (Zouboulis et al. 2015).

[0008] While topical antibiotics may be used for mild cases, for moderate to severe HS a long course of multidrug systemic antimicrobial therapy, generally with a combination of tetracycline or clindamycin and rifampicin, is preferred, which may be followed by maintenance on chronic antibiotic treatment for months or even years (Bettoli et al. 2016; Dessinioti et al. 2016; Zouboulis et al. 2015).

[0009] However, it is widely recognized that HS is a chronic inflammatory state and not an infectious disease (Jemec 2012). Anti-inflammatory agents may therefore be an alternative and perhaps more appropriate approach than or complementary to antibiotics. Over time, if chronic recurrent inflammation is inappropriately treated, it results in irreversible fibrosis, which manifests as scarring and tunnels or sinus tracts and is often unresponsive to drug therapy. Once persistent anatomical changes occur, surgery becomes the only treatment option to reduce the volume of fibrotic tissue and improve function in areas of affected skin (Andersen and Jemec 2017). One of the goals of future treatment should be to reduce residual scarring and avoid surgery, which may be achieved by prevention of inflammatory lesions or may require specific procedures.

[0010] In 2015, adalimumab (Humira®), a recombinant human monoclonal immunoglobulin G1 (IgG1) antibody against soluble and membrane-bound TNF-α, received regulatory approval for the treatment of moderate to severe HS. Efficacy has been seen with adalimumab, resulting in response rates over placebo of approximately 16% (41.8% adalimumab vs. 26% placebo) and 31% (58.9% adalimumab vs. 27.6% placebo) in HiSCR (hidradenitis suppurativa clinical response), as reported in the PIONEER I and II studies, respectively (Kimball et al. 2016). When captured by adalimumab labeling, adalimumab is associated with increased safety risks for serious infections, including tuberculosis, invasive fungal infections, and other opportunistic infections. Increased incidence of malignancies has also been reported with adalimumab.

[0011] Thus, there is an unmet need for systemic treatments that effectively reduce inflammation while having a favorable safety profile for patients with moderate to severe HS. Summary of the Invention

[0012] In HS, both cytokines IL-1β and IL-18 are upregulated (Kelly et al. 2015) and therefore may be involved in its pathogenesis. An IL-1β signature is present in HS lesions and can be reversed by application of IL-1 receptor antagonists (Witte-Handel et al. 2019). Anakinra, a recombinant IL-1R antagonist, has shown promising clinical efficacy results compared to placebo in small trials (Tzanetakou et al. 2016), while case reports have confirmed these findings (Leslie et al. 2014, Zarchi et al. 2013, Andre et al. 2019). Case reports have shown that IL-1β blockade using only the anti-IL-1β antibody canakinumab may be beneficial in moderate to severe HS (Houriet et al. 2017) or for related syndromes such as PASH (pyoderma gangrenosum, acne and hidradenitis suppurativa) (Jaeger et al. 2013). However, the observed partial failure in HS with canakinumab (Sun et al. 2017; Tekin et al. 2017) and anakinra (van der Zee and Prens 2013; Russo and Alikhan 2016) suggests that perhaps only a portion of the population may be responsive to anti-IL-1 blockade or that this blockade alone may not be sufficient to achieve results in most patients.

[0013] Thus, it is an object of the present disclosure to target both inflammasome effector cytokines, IL-1β and IL-18, and thus provide superior clinical efficacy in (auto)-inflammatory conditions, such as HS, or when both IL-1β and IL-18 contribute independently to the pathophysiology of the disease. A further object of the present disclosure is that a bispecific anti-IL-1β / 18 antagonist can rapidly neutralize inflammasome-dependent as well as inflammasome-independent sources of IL-1β and IL-18.

[0014] Therefore, any antagonist capable of inhibiting both IL-1β and IL-18, such as a bispecific anti-IL-1β / 18 antibody or a fragment thereof, may be suitable for the treatment of HS.

[0015] Described herein is a bispecific antibody or functional fragment thereof that simultaneously targets both IL-1β and IL-18 for use in preventing or treating hidradenitis suppurativa (HS) in a subject. In a preferred embodiment, the anti-IL-1β / 18 antibody comprises a heavy chain CH3 mutation that suppresses ADCC activity, such as the so-called LALA mutation, which comprises L234A and L235A mutations in the IgG1 Fc amino acid sequence. In a preferred embodiment, the anti-IL-1β / 18 antibody comprises a first heavy chain with complementary heavy chain CH3 knob-in-hole mutations, e.g., S354C and T366W (according to EU numbering), knob-type mutations, and a second heavy chain with Y349C, T366S, L368A, Y407V, hole-type mutations. In a preferred embodiment, the anti-IL-1β / 18 antibody comprises a first light chain that preferentially associates with the first heavy chain, the first light chain being a kappa light chain, e.g., Vκ6, and a second light chain that preferentially associates with the second heavy chain, the second light chain being a lambda light chain, e.g., Vλ1. In an even more preferred embodiment, the anti-IL-1β / 18 antibody comprises i) one or more ADCC silencing mutations (e.g., LALA), ii) one or more knob-in-hole heavy chain modifications, iii) and / or a combination of kappa and lambda light chains. In an even more preferred embodiment, the anti-IL-1β / 18 antibody comprises all of the aforementioned characteristics i)-iii), preferably the antibody comprises Vκ6 and Vλ1 light chains.

[0016] Also described herein are methods of preventing or treating hidradenitis suppurativa (HS) by administering to a subject in need thereof a therapeutically effective amount of a bispecific antibody that simultaneously targets both IL-1β and IL-18.

[0017] Further provided herein are specific dosing regimens for the methods or uses of bispecific antibodies (eg, bbmAb1) that simultaneously target both IL-1β and IL-18 as described herein.

[0018] Further described herein are pharmaceutical combinations and compositions comprising a) a bispecific antibody (e.g., bbmAb1) that simultaneously targets both IL-1β and IL-18, and b) at least one further therapeutic agent, optionally in the presence of a pharma- ceutical acceptable carrier, for use in the treatment or prevention of HS. Further properties and advantages of the described methods and uses will become apparent from the following detailed description.

[0019] In a first aspect, the present disclosure relates to a method for the treatment or prevention of HS in a subject in need thereof comprising administering to said subject a therapeutically effective amount of a bispecific antibody, the antibody comprising: a. a first moiety that is an immunoglobulin with a first variable light chain (VL1) and a first variable heavy chain (VH1) that specifically binds to IL1β, and a first constant heavy chain (CH1) with a heterodimerization modification; b. a second moiety that is an immunoglobulin with a second variable light chain (VL2) and a second variable heavy chain (VH2) that specifically binds IL-18, and a second constant heavy chain (CH2) with a heterodimerization modification that is complementary to the heterodimerization modification of the first constant heavy chain; Includes.

[0020] In a second aspect, the disclosure relates to a method for slowing, inhibiting, or reducing the onset of HS in a subject in need thereof comprising administering to said subject a therapeutically effective amount of a bispecific antibody, the antibody comprising: a. a first moiety that is an immunoglobulin with a first variable light chain (VL1) and a first variable heavy chain (VH1) that specifically binds to IL1β, and a first constant heavy chain (CH1) with a heterodimerization modification; b. a second moiety that is an immunoglobulin with a second variable light chain (VL2) and a second variable heavy chain (VH2) that specifically binds IL-18, and a second constant heavy chain (CH2) with a heterodimerization modification that is complementary to the heterodimerization modification of the first constant heavy chain; In certain embodiments of the first and second aspects of the disclosure, the first and second constant heavy chains of the bispecific antibody are human IgA, IgD, IgE, IgG or IgM, preferably IgD, IgE or IgG, such as human IgG1, IgG2, IgG3 or IgG4, preferably IgG1.

[0021] In another embodiment of the present disclosure, the first and second constant heavy chains of the bispecific antibody are IgG1; a. the first constant heavy chain has a point mutation that generates a knob structure and the second constant heavy chain has a point mutation that generates a hole structure, or b. the first constant heavy chain has a point mutation that generates a hole structure and the second constant heavy chain has a point mutation that generates a knob structure, and optionally c. The first and second constant heavy chains have mutations that create disulfide bridges.

[0022] In a particularly preferred embodiment of the present disclosure, including the first and second aspect, the first immunoglobulin VH1 domain of the bispecific antibody comprises i. hypervariable regions CDR1, CDR2 and CDR3, wherein CDR1 has the amino acid sequence SEQ ID NO: 76, CDR2 has the amino acid sequence SEQ ID NO: 77 and CDR3 has the amino acid sequence SEQ ID NO: 78; or ii. hypervariable regions CDR1, CDR2 and CDR3, wherein CDR1 has the amino acid sequence SEQ ID NO: 79, CDR2 has the amino acid sequence SEQ ID NO: 80 and CDR3 has the amino acid sequence SEQ ID NO: 81. Includes; The first immunoglobulin VL1 domain of the bispecific antibody comprises iii. hypervariable regions CDR1, CDR2 and CDR3, wherein CDR1 has the amino acid sequence SEQ ID NO: 92, CDR2 has the amino acid sequence SEQ ID NO: 93 and CDR3 has the amino acid sequence SEQ ID NO: 94; or iv. Hypervariable regions CDR1, CDR2 and CDR3, wherein CDR1 has the amino acid sequence SEQ ID NO: 95, CDR2 has the amino acid sequence SEQ ID NO: 96 and CDR3 has the amino acid sequence SEQ ID NO: 97. Includes; The second immunoglobulin VH2 domain of the bispecific antibody comprises v. hypervariable regions CDR1, CDR2 and CDR3, wherein CDR1 has the amino acid sequence SEQ ID NO: 44, CDR2 has the amino acid sequence SEQ ID NO: 45 and CDR3 has the amino acid sequence SEQ ID NO: 46; or vi. Hypervariable regions CDR1, CDR2 and CDR3, wherein CDR1 has the amino acid sequence SEQ ID NO: 47, CDR2 has the amino acid sequence SEQ ID NO: 48 and CDR3 has the amino acid sequence SEQ ID NO: 49. Includes; The second immunoglobulin VL2 domain of the bispecific antibody comprises vii. hypervariable regions CDR1, CDR2 and CDR3, wherein CDR1 has the amino acid sequence SEQ ID NO: 60, CDR2 has the amino acid sequence SEQ ID NO: 61 and CDR3 has the amino acid sequence SEQ ID NO: 62; or viii. Hypervariable regions CDR1, CDR2 and CDR3, wherein CDR1 has the amino acid sequence SEQ ID NO: 63, CDR2 has the amino acid sequence SEQ ID NO: 64 and CDR3 has the amino acid sequence SEQ ID NO: 65. Includes.

[0023] In another preferred embodiment of the present disclosure, the antibody used in the methods, compositions and uses described herein is a. a first immunoglobulin VH1 domain of the amino acid sequence SEQ ID NO: 85; b. A first immunoglobulin VL1 domain of the amino acid sequence SEQ ID NO: 101; c. a second immunoglobulin VH2 domain of the amino acid sequence SEQ ID NO:53; and d. A second immunoglobulin VL2 domain of the amino acid sequence SEQ ID NO:69 Includes.

[0024] In another preferred embodiment of the present disclosure, the antibody used in the methods, compositions or uses described herein is e. a first immunoglobulin heavy chain of the amino acid sequence SEQ ID NO: 87; f. a first immunoglobulin light chain of the amino acid sequence SEQ ID NO: 103; g. a second immunoglobulin heavy chain of the amino acid sequence SEQ ID NO:55; and h. A second immunoglobulin light chain having the amino acid sequence SEQ ID NO:71 Includes.

[0025] In a particularly preferred embodiment of the present disclosure, the subject being treated has HS.

[0026] In a third aspect, the disclosure provides a method for treating or preventing HS in a subject in need thereof, comprising: a. a first moiety that is an immunoglobulin with a first variable light chain (VL1) and a first variable heavy chain (VH1) that specifically binds to IL1β, and a first constant heavy chain (CH1) with a heterodimerization modification; b. a second moiety that is an immunoglobulin with a second variable light chain (VL2) and a second variable heavy chain (VH2) that specifically binds IL-18, and a second constant heavy chain (CH2) with a heterodimerization modification that is complementary to the heterodimerization modification of the first constant heavy chain; The present invention relates to a bispecific antibody comprising:

[0027] In a fourth aspect, the disclosure relates to the methods and treatments of the first, second and third aspects, wherein about 1 mg / kg to about 35 mg / kg of a bispecific antibody that simultaneously targets both IL-1β and IL-18 is administered to the subject. In a preferred embodiment of the fourth aspect, about 10 mg / kg of the bispecific antibody is administered to the treated subject.

[0028] In one aspect of the disclosure, a bispecific antibody that simultaneously targets both IL-1β and IL-18 is administered to a subject intravenously or subcutaneously. In one embodiment, the route of administration is a combination of subcutaneous or intravenous, such as an intravenous loading dose followed by subcutaneous maintenance doses.

[0029] In one embodiment, a bispecific antibody that simultaneously targets both IL-1β and IL-18 is administered to a subject to be treated, e.g., intravenously, at a dose of about 1.5 mg to about 15 mg of active ingredient / kilogram human subject. In one embodiment, a bispecific antibody that simultaneously targets both IL-1β and IL-18 is administered to a subject to be treated, e.g., intravenously, at a dose of about 2, 4, or 5 mg of active ingredient / kilogram human subject. In one embodiment, a bispecific antibody that simultaneously targets both IL-1β and IL-18 is administered to a subject to be treated, e.g., intravenously, at a dose of about 4 mg of active ingredient / kilogram human subject. For example, a dose of about 1.5 to about 15 mg of active ingredient / kilogram human subject may be given weekly, every two weeks, or every four weeks, or a combination thereof. In a preferred embodiment, for example, a dose of about 1.5 to about 15 mg of active ingredient / kilogram human subject may be given every two weeks, or every four weeks, or a combination thereof.

[0030] In one embodiment, the dose is about 75 mg to about 600 mg of active ingredient, preferably about 150 mg to about 300 mg of active ingredient, or about 150 mg or 300 mg of active ingredient, preferably 300 mg of active ingredient. The dose may be given weekly, every two weeks, or every four weeks, or a combination thereof. In a preferred embodiment, the fixed dose (e.g., non-weight-based dose) is administered subcutaneously or intramuscularly, preferably subcutaneously.

[0031] In one embodiment, the dosage is about 300 mg of active ingredient.

[0032] In one preferred embodiment, the dose is 150 mg of active ingredient. In another preferred embodiment, the dose is 300 mg of active ingredient. In yet another preferred embodiment, the dose is 600 mg of active ingredient. In yet another embodiment, the dose is from about 150 mg of active ingredient to about 600 mg of active ingredient.

[0033] In one embodiment, the antibody is administered via a loading dose and a maintenance dose. In one embodiment, the loading dose is administered via one or more intravenous injections of a first dose, and the maintenance dose is administered via subcutaneous injections of a second dose. In one embodiment, the loading dose is administered via subcutaneous injection of a first dose, and the maintenance dose is administered via subcutaneous injection of a second dose.

[0034] In one embodiment, the loading dose is administered via subcutaneous injection of a first dosing regimen, and the maintenance dose is administered via subcutaneous injection of a second dosing regimen. The amount of the first dosing regimen can be the same as or greater than the amount of the second dosing regimen. The duration of the first dosing regimen can be the same as or more frequent than the duration of the second dosing regimen.

[0035] In one embodiment, the first dose is from about 150 mg to about 600 mg of active ingredient, such as about 300 mg of active ingredient, and the second dose is from about 150 mg to about 600 mg of active ingredient, such as about 300 mg of active ingredient.

[0036] In one embodiment, the first dose is 150 mg, 300 mg, or 600 mg of active ingredient and the second dose is 150 mg, 300 mg, or 600 mg of active ingredient.In one embodiment, the first dose is 300 mg of active ingredient and the second dose is 300 mg of active ingredient.

[0037] In one embodiment, the loading dose comprises at least two administrations and the maintenance doses consist of weekly (Q1W), preferably biweekly (Q2W) or monthly (Q4W) administrations. In one embodiment, the loading dose consists of two administrations. In another embodiment, the loading dose consists of three administrations. In another embodiment, the loading dose consists of four administrations. In some cases, the loading dose is a biweekly loading dose consisting of three administrations on days 1 through 29.

[0038] In one embodiment, the loading dose comprises at least two subcutaneous injections, preferably three subcutaneous injections, and the maintenance dose comprises weekly (Q1W), biweekly (Q2W) or preferably monthly (Q4W) subcutaneous injections. In one embodiment, the loading dose comprises two subcutaneous injections. In another embodiment, the loading dose comprises three subcutaneous injections, e.g., three 150 mg or 300 mg subcutaneous injections every other week (Q2W). In another embodiment, the loading dose comprises four subcutaneous injections. In some cases, the loading dose is a biweekly loading dose consisting of three subcutaneous injections administered on days 1-29. In some cases, the maintenance dose is a monthly (Q4W) maintenance dose including a subcutaneous Q4W injection administration starting on day 57, e.g., three biweekly (Q2W) 150 mg or 300 mg subcutaneous injections. In some cases, the loading dose is a biweekly loading dose consisting of three subcutaneous injections administered on days 1, 15, and 29, and the maintenance dose is a monthly (Q4W) maintenance dose with a subcutaneous Q4W injection dose starting on day 57. In some cases, the loading dose is a biweekly (e.g., 150 mg or 300 mg) loading dose consisting of three subcutaneous injections administered on days 1, 15, and 29, and the maintenance dose is a monthly (Q4W) (e.g., 150 mg or 300 mg) maintenance dose with a subcutaneous Q4W injection dose starting on day 57.

[0039] In one embodiment, the loading dose is a biweekly loading dose consisting of three administrations on days 1 through 29, where the dosage is about 1.5 mg to about 15 mg / kilogram of active ingredient, for example, where the loading dose is administered intravenously, subcutaneously, or a combination of intravenous or subcutaneous. In one embodiment, the loading dose is a biweekly loading dose consisting of three administrations on days 1 through 29, where the dosage is about 4 mg / kilogram of active ingredient, for example, where the loading dose is administered intravenously, subcutaneously, or a combination of intravenous or subcutaneous.

[0040] In one embodiment, the loading dose is a biweekly loading dose consisting of three administrations on days 1-29, the dosage being about 150 mg or about 300 mg of active ingredient, preferably 300 mg of active ingredient, e.g., the loading dose is administered subcutaneously or a combination of intravenous or subcutaneous. In one embodiment, the loading dose is a biweekly loading dose consisting of three administrations on days 1-29, the dosage being about 300 mg of active ingredient, and the loading dose is administered subcutaneously.

[0041] In some embodiments, the initial loading dose is a dose selected or expected to achieve a plasma concentration of at least about 20 μg / mL to about 60 μg / mL during the initial loading dosing period (e.g., within 1, 2, or 3 weeks or after 1, 2, or 3 initial loading dose injections). In some embodiments, the maintenance dose is a dose selected or expected to provide a sustained plasma concentration of about 20 μg / mL to about 60 μg / mL during the maintenance dosing period, or a substantial portion thereof (e.g., at least about day 29 to about day 85).

[0042] In one embodiment, the initial loading subcutaneous injections are of different doses, hi another embodiment, the initial loading subcutaneous injections are of the same dose.

[0043] HS patients may be selected according to one of the following criteria: The patient has moderate to severe HS; The patient is an adult; The patient is an adolescent; the patient has an HS-PGA score of ≥3 prior to treatment with an IL-1β and IL-18 antagonist (e.g., a bispecific antibody targeting both IL-1β and IL-18); The patient has at least three inflammatory lesions prior to treatment with an IL-1β and IL-18 antagonist (e.g., a bispecific antibody that targets both IL-1β and IL-18); or Prior to treatment with IL-1β and IL-18 antagonists (e.g., a bispecific antibody targeting both IL-1β and IL-18), patients had no extensive scarring (<10 fistulas) as a result of HS.

[0044] In one embodiment, by week 16 of treatment, the HS patient achieves at least one of the following: Simple HiSCR; Reduction of HS erythema; NRS30; A decrease of ≦6 as measured by the DLQI; and / or Improved DLQI.

[0045] In one embodiment, by week 16 of treatment, at least 40% of the patients achieve a simplified HiSCR; or at least 25% of the patients achieve an NRS30 response; or less than 15% of the patients experience HS erythema.

[0046] In one embodiment, the patient, as early as one week after the first dose of the IL-1β and IL-18 antagonist, has at least one of the following: Rapid relief of pain as measured by VAS or NRS, and A rapid decline in CRP as measured using a standard hsCRP assay.

[0047] In one embodiment, the patient achieves a sustained response three months after the end of treatment as measured by inflammatory lesion counts, HS Clinical Response (HiSCR), Numeric Rating Scale (NRS), Modified Sartorius HS score, Hidradenitis Suppurativa-Physician Global Assessment (HS-PGA) or Dermatology Life Quality Index (DLQI).

[0048] In one embodiment, the patient achieves a sustained response as measured by simple HiSCR (sHiSCR) three months after the end of treatment.

[0049] According to a second aspect, there is provided a method of treating HS in a human subject comprising administering to the human subject a therapeutically effective dose of an IL-1β and an IL-18 antagonist.

[0050] In another embodiment of the present disclosure, a bispecific antibody targeting both IL-1β and IL-18 is administered in combination with at least one additional therapeutic agent.

[0051] In another aspect, the disclosure relates to the use of a bispecific antibody that targets both IL-1β and IL-18 (e.g., bbmAb1) for the manufacture of a medicament for treating, slowing, arresting, or alleviating the onset of HS in a subject.

[0052] In one embodiment, provided herein is a method of treating HS in a subject in need thereof, comprising administering an effective amount of a bispecific antibody that targets both IL-1β and IL-18, e.g., bbmAb1. In one embodiment, provided herein is a bispecific antibody that targets both IL-1β and IL-18, e.g., bbmAb1, for use in treating HS in a subject in need thereof. In some embodiments, provided herein is the use of a bispecific antibody that targets both IL-1β and IL-18, e.g., bbmAb1, for the manufacture of a medicament for treating HS.

[0053] In one embodiment, provided herein is a method for treating, slowing, arresting or reducing the onset of HS in a subject in need thereof, comprising administering to said subject a therapeutically effective amount of a bispecific antibody that targets both IL-1β and IL-18, e.g., bbmAb1. In one embodiment, the therapeutically effective amount is an amount of a bispecific antibody that comprises: a. the first immunoglobulin VH1 domain of the bispecific antibody comprises the amino acid sequence SEQ ID NO:85; b. the first immunoglobulin VL1 domain of the bispecific antibody comprises the amino acid sequence SEQ ID NO:101; c. the second immunoglobulin VH2 domain of the bispecific antibody comprises the amino acid sequence SEQ ID NO:53; d. The second immunoglobulin VL2 domain of the bispecific antibody comprises the amino acid sequence SEQ ID NO:69.

[0054] According to another aspect, the disclosure provides a liquid pharmaceutical composition, a method or use of the liquid pharmaceutical composition comprising a bispecific antibody that specifically targets both IL-18 and IL-1β, the liquid pharmaceutical formulation comprising a buffer, a stabilizer and a solubilizer. In one embodiment, the disclosure provides or further provides a means for administering or subcutaneously administering the bispecific antibody to a patient with HS. In one embodiment, the use is for the manufacture of a medicament for the treatment of HS.

[0055] In a particularly preferred embodiment of the present disclosure, the first immunoglobulin VH1 domain of the bispecific antibody of the liquid pharmaceutical composition comprises: i. hypervariable regions CDR1, CDR2 and CDR3, wherein CDR1 has the amino acid sequence SEQ ID NO: 76, CDR2 has the amino acid sequence SEQ ID NO: 77 and CDR3 has the amino acid sequence SEQ ID NO: 78; or ii. hypervariable regions CDR1, CDR2 and CDR3, wherein CDR1 has the amino acid sequence SEQ ID NO: 79, CDR2 has the amino acid sequence SEQ ID NO: 80 and CDR3 has the amino acid sequence SEQ ID NO: 81. Includes; The first immunoglobulin VL1 domain of the bispecific antibody comprises iii. hypervariable regions CDR1, CDR2 and CDR3, wherein CDR1 has the amino acid sequence SEQ ID NO: 92, CDR2 has the amino acid sequence SEQ ID NO: 93 and CDR3 has the amino acid sequence SEQ ID NO: 94; or iv. Hypervariable regions CDR1, CDR2 and CDR3, wherein CDR1 has the amino acid sequence SEQ ID NO: 95, CDR2 has the amino acid sequence SEQ ID NO: 96 and CDR3 has the amino acid sequence SEQ ID NO: 97. Includes; The second immunoglobulin VH2 domain of the bispecific antibody comprises v. hypervariable regions CDR1, CDR2 and CDR3, wherein CDR1 has the amino acid sequence SEQ ID NO: 44, CDR2 has the amino acid sequence SEQ ID NO: 45 and CDR3 has the amino acid sequence SEQ ID NO: 46; or vi. Hypervariable regions CDR1, CDR2 and CDR3, wherein CDR1 has the amino acid sequence SEQ ID NO: 47, CDR2 has the amino acid sequence SEQ ID NO: 48 and CDR3 has the amino acid sequence SEQ ID NO: 49. Includes; The second immunoglobulin VL2 domain of the bispecific antibody comprises vii. hypervariable regions CDR1, CDR2 and CDR3, wherein CDR1 has the amino acid sequence SEQ ID NO: 60, CDR2 has the amino acid sequence SEQ ID NO: 61 and CDR3 has the amino acid sequence SEQ ID NO: 62; or viii. Hypervariable regions CDR1, CDR2 and CDR3, wherein CDR1 has the amino acid sequence SEQ ID NO: 63, CDR2 has the amino acid sequence SEQ ID NO: 64 and CDR3 has the amino acid sequence SEQ ID NO: 65. Includes.

[0056] In another preferred embodiment of the present disclosure, the antibody of the liquid pharmaceutical composition described herein comprises: a. a first immunoglobulin VH1 domain of the amino acid sequence SEQ ID NO: 85; b. A first immunoglobulin VL1 domain of the amino acid sequence SEQ ID NO: 101; c. a second immunoglobulin VH2 domain of the amino acid sequence SEQ ID NO:53; and d. A second immunoglobulin VL2 domain of the amino acid sequence SEQ ID NO:69 Includes.

[0057] In another preferred embodiment of the present disclosure, the antibody of the liquid pharmaceutical composition described herein comprises: e. a first immunoglobulin heavy chain of the amino acid sequence SEQ ID NO: 87; f. a first immunoglobulin light chain of the amino acid sequence SEQ ID NO: 103; g. a second immunoglobulin heavy chain of the amino acid sequence SEQ ID NO:55; and h. A second immunoglobulin light chain having the amino acid sequence SEQ ID NO:71 Includes.

[0058] In some embodiments, the liquid pharmaceutical composition comprises about 50 mg / mL to about 120 mg / mL of the bispecific antibody (e.g., bbmAb1), a buffer, a stabilizer (e.g., a non-ionic stabilizer such as a sugar), and a surfactant (e.g., polysorbate 20 or polysorbate 80). In some embodiments, the liquid pharmaceutical composition comprises about 50 mg / mL to about 120 mg / mL of the bispecific antibody in a buffer at a pH of about 5.5 to about 6.5, preferably at a pH of about 5.5 to about 6.0, preferably at a pH of about 6.0.

[0059] In some embodiments, the liquid pharmaceutical composition comprises about 100 mg / mL of the bispecific antibody (e.g., bbmAb1), a buffer, a stabilizer (e.g., a non-ionic stabilizer, such as a sugar), and a surfactant (e.g., polysorbate 20 or polysorbate 80). In some embodiments, the liquid pharmaceutical composition comprises about 50 mg / mL to about 120 mg / mL of the bispecific antibody (e.g., bbmAb1), about 1 mM to about 50 mM histidine / histidine-chloride, about 50 mM to about 400 mM sugar (e.g., sucrose, trehalose, or mannitol), about 0.01% to about 0.5% surfactant (e.g., a polysorbate, such as polysorbate 20), at a pH of about 5.5 to about 6.5, preferably about 5.5 to about 6.0, more preferably about 6.0. In some cases, the liquid pharmaceutical composition comprises about 50 mg / mL to about 120 mg / mL of bbmAb1 (preferably about 100 mg / mL of bbmAb1), about 20 mM histidine / histidine-chloride, about 220 mM sucrose, about 0.04% polysorbate 20 (w / v), at a pH of about 6.0. [Brief description of the drawings]

[0060] [Figure 1] FIG. 1 is a graphical representation of AIM2, NLRC4, NLRP7 and NLRP3 inflammasome mRNA levels in HS lesions compared to levels in non-lesioned or healthy tissue. [Diagram 2] Figure 2 is a TIBCO spotfire heatmap depiction of the mean expression levels of IL-18, IL-12 and IL-1β genes / members (alone or in combination) or CRS signaling signatures in healthy skin or lesional, perilesional and lesional skin biopsies from HS patients. Identification of genes / members of individual IL-18, IL-12 and IL-1β signaling signatures was performed in separate experiments using PBMCs from healthy donors stimulated for 6 h (restricted to genes induced by individual cytokines = UP) or adopted from publications (canakinumab response signature CRS; Brachat et al., 2017). [Diagram 3]FIG. 3 shows the effect of either bbmAb1 (100 μg / mL) or adalimumab (100 μg / mL) incubation compared to untreated controls on the levels of the analytes IFNγ, TNFα, IL-1β and IL-2 produced in the supernatants of HS skin biopsies after 24 hours of in vitro culture. Each dot represents one biopsy. The leftmost column for each specimen is the untreated control, the middle column for each specimen is bbmAb1, and the rightmost column for each specimen is adalimumab. Values ​​are shown from n=54, n=36 and n=26 individual biopsies for IFNγ, IL-1β and TNFα, and n=43, n=20 and n=10 biopsies for IL-2 from 9 different HS patients. [Figure 4] Figure 4 is a graphical representation of a prediction of the effect of bbmAb1 administration on IL-1β levels. The dotted line indicates the lower limit of quantification. The predictions are modeled assuming subcutaneous administration of 300 mg on days 1, 15, 29, 57 and 85. [Diagram 5] Figure 5 is a graphical representation of the prediction of the effect of bbmAb1 administration on IL-18 levels versus baseline change in healthy control subjects. The dotted line indicates the levels in healthy control subjects. Predictions are modeled assuming subcutaneous administration of 300 mg on days 1, 15, 29, 57 and 85. [Figure 6] Figure 6 is a graphical representation of the predicted pharmacokinetics of subcutaneously administered bbmAb1 compared to a single dose of 10 mg / kg iv. Predictions are modeled assuming subcutaneous administration of 300 mg on days 1, 15, 29, 57 and 85. [Figure 7] FIG. 7 shows the results of a test on aggregate formation. [Figure 8] FIG. 8 shows the results of a study of degradation product formation. [Figure 9] FIG. 9 shows the results of a study of acidic and basic variant formation. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0061] Described herein is an IgG-like bispecific monoclonal antibody (e.g., bbmAb1) that contains a heterodimeric Fc portion that simultaneously neutralizes the key inflammasome effector cytokines interleukin-1 beta (IL-1β) and interleukin-18 (IL-18). In certain embodiments, the bispecific antibody has picomolar (pM) affinity for both cytokines and inhibits IL-1β and IL-18 signaling at subnanomolar (nM) concentrations in cellular in vitro assays. IL-1β and IL-18 are two pro-inflammatory cytokines secreted following inflammasome activation in response to the recognition of damage-associated molecular patterns (DAMPs) and pathogen-associated molecular patterns (PAMPs). As described herein, combined inhibition of IL-1β and IL-18 can result in more effective down-regulation of inflammasome-driven pro-inflammatory responses compared to inhibition of either cytokine alone.

[0062] Therefore, any antagonist capable of simultaneously blocking IL-1β and IL-18 activity, such as a bispecific anti-IL-1β and anti-IL-18 antibody with suppressed ADCC activity, may be suitable for the treatment of HS.

[0063] Without wishing to be bound by theory, the inventors have identified that a loading regimen may be beneficial at the beginning of treatment and that a higher dose or more frequent dosing regimen is required at the beginning of treatment to at least partially or completely saturate the bispecific antibody binding sites (IL-1β and / or IL-18) in these patients in conditions where IL-1β and / or IL-18 levels are promoted. Thus, it is considered for therapeutic efficacy that a loading regimen provides rapid saturation of the antigen at the beginning of treatment (2-3 weeks), followed by a maintenance regimen to provide sustained therapeutic plasma concentrations throughout the treatment period in situations where IL-1β and / or IL-18 expression in affected tissues is promoted (severity of condition).

[0064] Suitable dosages will vary depending, for example, on the particular bispecific IL-1β and IL-18 antagonist, such as the bispecific anti-IL-1β and anti-IL-18 antibody or antigen-binding fragment thereof (e.g., bbmAb1) to be used, the subject of treatment, the mode of administration, and the nature and severity of the condition being treated, and on the nature of previous treatments the patient has undergone. Ultimately, the responsible healthcare provider will determine the amount of bispecific IL-1β and IL-18 antagonist with which to treat each individual patient. In some embodiments, the responsible healthcare provider may administer a low or equal single dose of the bispecific IL-1β and IL-18 antagonist and observe the patient's response. In other embodiments, the patient is administered a high initial dose of the bispecific IL-1β and IL-18 antagonist, which is then tapered until signs of relapse occur. Higher doses of the bispecific IL-1β and IL-18 antagonist may be administered until an optimal therapeutic effect is achieved for the patient, and the dosage is generally not further increased.

[0065] In one embodiment, the present disclosure relates to a bispecific anti-IL-1β and anti-IL-18 antibody or antigen-binding fragment thereof for use according to any of the aspects or embodiments of the present disclosure as described above, wherein the loading dose and maintenance dose of the bispecific anti-IL-1β and anti-IL-18 antibody or antigen-binding fragment thereof (e.g., bbmAb1) are adjusted to achieve therapeutic plasma or serum concentrations of the antibodies.

[0066] In some implementations of the disclosed methods of treatment or use, a therapeutically effective amount of a bispecific IL-1β and IL-18 antagonist, such as a bispecific anti-IL-1β and anti-IL-18 antibody or antigen-binding fragment thereof (e.g., bbmAb1), is administered to a patient, such as a mammal (e.g., a human). While it is understood that the disclosed methods provide for the treatment of HS patients with a bispecific IL-1β and IL-18 antagonist (e.g., bbmAb1), this does not exclude that such treatment is necessarily a monotherapy if the patient is ultimately to be treated with the antagonist. Indeed, if a patient is selected for treatment with a bispecific IL-1β and IL-18 antagonist, the antagonist (e.g., bbmAb1) may be administered according to the disclosed methods either alone or in combination with other drugs and treatments.

[0067] It will be appreciated that for certain HS patients, such as those who respond inadequately to treatment with a bispecific IL-1β and IL-18 antagonist, such as a bispecific antibody or antigen-binding fragment thereof (e.g., bbmAb1), modified dosing regimens may be appropriate. Thus, dosing may be more frequent than monthly, such as biweekly (every two weeks) or weekly.

[0068] Some patients may benefit from a loading regimen (e.g., weekly dosing for several weeks (e.g., 1-4 weeks, e.g., dosing at weeks 0, 1, 2 and / or 3, e.g., a loading regimen at weeks 1, 2 and 3, e.g., 3 weeks) followed by a maintenance regimen beginning at, e.g., week 5, 6 or 7, in which the bispecific antibody (e.g., bbmAb1) may be administered weekly, every other week, or preferably every 4 weeks for at least several weeks.

[0069] For example, a suitable dosing regimen for bbmAb1 may be every other week for several weeks [e.g., dosing for weeks 1-5, e.g., weeks 1, 3 and 5] followed by a maintenance dosing regimen of Q4W every month, e.g., in weeks 8 or 9, preferably week 9, where the bispecific antibody (e.g., bbmAb1) can be administered for at least several weeks.

[0070] It will also be appreciated that administration can be less frequent than monthly, such as every 6 weeks, every 8 weeks (every 2 months), four times a year (every 3 months), etc.

[0071] It will be appreciated that dose escalation may be appropriate for certain HS patients, for example those who respond poorly to treatment with a bispecific antagonist described herein, such as bbmAb1 or an antigen-binding fragment thereof, based on the severity of the disease. Thus, subcutaneous (sc) doses (loading or maintenance doses) may be greater than about 150 mg to about 900 mg sc, such as about 75 mg, about 100 mg, about 125 mg, about 175 mg, about 200 mg, about 250 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 600 mg, etc.; similarly, intravenous (iv) doses may be greater than about 5 mg / kg or 10 mg / kg, such as about 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, 11 mg / kg, 12 mg / kg, 15 mg / kg, 20 mg / kg, 25 mg / kg, 30 mg / kg, 35 mg / kg, etc. It will also be appreciated that dose reductions may be appropriate for certain HS patients, such as those who experience adverse events or reactions to treatment with a bispecific antagonist (e.g., bbmAb1 or an antigen-binding fragment thereof). Thus, the dosage of the antagonist may be from about 150 mg to less than about 900 mg sc, such as about 25 mg, about 50 mg, about 75 mg, about 100 mg, about 125 mg, about 175 mg, about 200 mg, about 250 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 600 mg, etc.

[0072] In some embodiments, the bispecific antagonist, such as bbmAb1 or an antigen-binding fragment thereof, may be administered to a patient at an initial dose of 300 mg delivered sc, and then the dose may be adjusted to 150 mg or 600 mg delivered sc as needed, as determined by the physician.

[0073] The bispecific antibody or antigen-binding fragment thereof may be bbmAb1, a functional derivative thereof or a bioanalog thereof.

[0074] As defined herein, a "unit dose" refers to a sc dose that may contain about 75 mg to about 900 mg, such as about 150 mg to about 600 mg, such as about 150 mg to about 600 mg, such as about 300 mg to about 600 mg, or for example about 150 mg to about 300 mg. For example, a unit sc dose may be about 75 mg, about 150 mg, about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 550 mg, about 600 mg.

[0075] The present disclosure is based inter alia on the unexpected finding that certain antibodies that simultaneously neutralize IL-1β and IL-18 more potently attenuate IFN-γ (and other pro-inflammatory cytokines) production compared to single IL-1β or IL-18 neutralization alone, which is believed by the inventors to be an effective treatment for HS.

[0076] 1.Definition For purposes of interpreting this specification, the following definitions will apply, and whenever appropriate, terms used in the singular will also include the plural and vice versa. Further definitions are set forth throughout the detailed description.

[0077] The term "about" with respect to a numerical value x means, for example, + / - 10%. When used before a range of numbers or a list of numbers, the term "about" applies to each number in the series, e.g., the phrase "about 1 to 5" should be interpreted as "about 1 to about 5", or, for example, the phrase "about 1, 2, 3, 4" should be interpreted as "about 1, about 2, about 3, about 4, etc."

[0078] The term "substantially" does not exclude "completely", e.g., a composition that is "substantially free" of Y may be completely free of Y. If desired, the term "substantially" may be omitted from the definition of the present disclosure.

[0079] The term "comprising" encompasses "comprising" as well as "consisting of," e.g., a composition "comprising" X may consist exclusively of X or may include some additional, e.g., X+Y.

[0080] The term "IL-18" is synonymous with IL-18 polypeptide, interleukin-18 polypeptide, IFN-gamma inducer or interferon-gamma-inducer or INF-γ inducer. The term "IL-18" refers to human IL-18, unless another species is specified. IL-18 is well known to those skilled in the art and is available, for example, from MBL® International Corporation under product number #B001-5. Throughout this application, the term IL-18 interchangeably encompasses both pro-IL-18 (the precursor of mature IL-18 before protease cleavage) and mature IL-18 (after protease cleavage), unless specified that the pro or mature form is meant.

[0081] The terms "IL-1β" or "IL-1b" are synonyms of IL-1β polypeptide and interleukin-1β polypeptide. The term "IL-1β" refers to human IL-1β, unless another species is specified. IL-1β is well known to those of skill in the art and is available, for example, from Sino Biological under product number #10139-HNAE-5.

[0082] The term "antibody" refers to an intact immunoglobulin or a functional fragment thereof. Natural antibodies generally comprise a tetramer, usually composed of at least two heavy (H) chains and at least two light (L) chains. Each heavy chain is composed of a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region, usually composed of three domains (CH1, CH2 and CH3). Heavy chains can be of any isotype, including IgG (IgG1, IgG2, IgG3 and IgG4 subtypes), IgA (IgA1 and IgA2 subtypes), IgM and IgE. Each light chain is composed of a light chain variable region (abbreviated herein as VL) and a light chain constant region (CL). Light chains include kappa (κ) and lambda (λ) chains. The heavy and light chain variable regions are generally responsible for antigen recognition, while the heavy and light chain constant regions may mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (Clq) of the classical complement system. The VH and VL regions may be further subclassified into regions of hypervariability called complementarity determining regions (CDRs) interspersed with more conserved regions called framework regions (FRs). Each VH and VL is composed of three CDRs and four FRs arranged from the amino terminus to the carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The heavy and light chain variable regions contain the binding domains that interact with antigens.

[0083] The term "antigen-binding portion" of an antibody (or simply "antigen portion"), as used herein, refers to a full-length or one or more fragments of an antibody that retains the ability to specifically bind to an IL-18 or IL-1β antigen. It has been shown that the antigen-binding function of an antibody can be performed by fragments of a full-length antibody. Examples of binding fragments encompassed within the term "antigen-binding portion" of an antibody include a Fab fragment, a monovalent fragment consisting of the VL, VH, CL and CH1 domains; a F(ab)2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; an Fd fragment consisting of the VH and CH1 domains; an Fv fragment consisting of the VL and VH domains of a single arm of an antibody; a dAb fragment consisting of the VH domain (Ward et al., (1989) Nature; 341:544-546); and an isolated complementarity determining region (CDR).

[0084] Furthermore, although the two domains of the Fv fragment, VL and VH, are encoded by separate genes, they can be linked using recombinant techniques by a flexible linker that allows them to be produced as a single protein chain in which the VL and VH regions pair to form a monovalent molecule (known as single-chain Fv (scFv); e.g., Bird et al., (1988) Science 242:423-426; and Huston et al., (1988) Proc Natl Acad Sc;.85:5879-5883). Such single-chain antibodies are also intended to be encompassed within the term "antigen-binding portion" of an antibody. These antibody fragments are obtained using conventional techniques known to those of skill in the art, and the fragments are screened for utility in the same manner as are intact antibodies.

[0085] The term "isolated" means throughout this application that an immunoglobulin, antibody or polynucleotide, as the case may be, may be present in a physical environment different from that in which it may occur in nature.

[0086] Throughout this application, complementarity determining regions ("CDRs") are defined according to the definition of Kabat, unless otherwise indicated. The precise amino acid sequence boundaries of a CDR can be determined using the Kabat numbering scheme, Kabat et al. (1991), "Sequences of Proteins of Immunological Interest," 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD ("Kabat" numbering scheme), Al-Lazikani et al., (1997) JMB 273, 927-948 ("Chothia" numbering scheme), and ImMunoGenTics (IMGT) numbering (Lefranc, M.-P., The Immunologist, 7, 132-136 (1999); Lefranc, M.-P. et al., Dev. Comp. Immunol., 27, 55-77 (2003) ("IMGT" numbering scheme). For example, in the classical format, under Kabat, the CDR amino acid residues in the heavy chain variable domain (VH) are numbered 31-35 (HCDR1), 50-65 (HCDR2) and 95-102 (HCDR3); and the CDR amino acid residues in the light chain variable domain (VL) are numbered 24-34 (LCDR1), 50-56 (LCDR2) and 89-97 (LCDR3). Under Chothia, the CDR amino acids in the VH are numbered 26-32 (HCDR1), 50-65 (HCDR2) and 95-102 (HCDR3). ), 52-56 (HCDR2) and 95-102 (HCDR3); amino acid residues in VL are numbered 26-32 (LCDR1), 50-52 (LCDR2) and 91-96 (LCDR3). Combining both Kabat and Chothia CDR definitions, the CDRs consist of amino acid residues 26-35 (HCDR1), 50-65 (HCDR2) and 95-102 (HCDR3) in human VH and amino acid residues 24-34 (LCDR1), 50-56 (LCDR2) and 89-97 (LCDR3) in human VL.Under IMGT, the CDR amino acid residues in VH are approximately 26-35 (CDR1), 51-57 (CDR2) and 93-102 (CDR3), and the CDR amino acid residues in VL are approximately 27-32 (CDR1), 50-52 (CDR2) and 89-97 (CDR3) (numbered according to "Kabat"). Under IMGT, the CDR regions of an antibody can be determined using the program IMGT / DomainGap Align.

[0087] By convention, the CDR regions in the heavy chain are commonly referred to as H-CDR1, H-CDR2 and H-CDR3, and in the light chain as L-CDR1, LCDR2 and L-CDR3, which are numbered sequentially from the amino terminus to the carboxy terminus.

[0088] The terms "monoclonal antibody" or "monoclonal antibody composition" as used herein refer to a preparation of antibody molecules of single molecular composition. A monoclonal antibody composition displays a single binding specificity and affinity for a particular epitope.

[0089] The term "human antibody," as used herein, is intended to include antibodies having variable regions in which both the framework and CDR regions are derived from sequences of human origin. Furthermore, if the antibody contains a constant region, the constant region may also be derived from such a human sequence, e.g., an antibody containing human germline sequences or mutated versions of human germline sequences or consensus framework sequences derived from human framework sequence analysis, e.g., as described in Knappik, et al., (2000) J Mol Biol; 296:57-86).

[0090] The human antibodies of the invention may include amino acid residues not encoded by human sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo). However, the term "human antibody", as used herein, is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, have been grafted onto human framework sequences.

[0091] The term "human monoclonal antibody" refers to antibodies displaying a single binding specificity which have variable regions in which both the framework and CDR regions are derived from human sequences.

[0092] The term "recombinant human antibody", as used herein, includes all human antibodies prepared, expressed, generated, or isolated by recombinant means, such as antibodies isolated from animals (e.g., mice) that are transgenic or transchromosomal for human immunoglobulin genes or hybridomas prepared therefrom, antibodies isolated from host cells that have been transformed to express human antibodies, e.g., from transfectomas, antibodies isolated from recombinant, combinatorial human antibody libraries, and antibodies prepared, expressed, generated, or isolated by any other means including splicing of all or a portion of a human immunoglobulin gene. Such recombinant human antibodies have variable regions in which the framework and CDR regions are derived from human germline immunoglobulin sequences. However, in certain embodiments, such recombinant human antibodies may be subjected to in vitro mutagenesis (or in vivo somatic mutagenesis, when animals transgenic for human Ig sequences are used) such that the amino acid sequences of the VH and VL regions of the recombinant antibodies, while derived from and related to human germline VH and VL sequences, are sequences that may not naturally occur within the in vivo human antibody germline repertoire.

[0093] The phrases "an antibody that recognizes an antigen" and "an antibody specific for an antigen" are used interchangeably herein with the term "an antibody that specifically binds to an antigen."

[0094] As used herein, a binding molecule that "specifically binds to IL-18" has a K D The term "IL-18 binding molecule" refers to a binding molecule that binds to human IL-18 at the

[0095] As used herein, a binding molecule that "specifically binds to IL-1β" has a K D The term "IL-1β binding molecule" refers to a binding molecule that binds to human IL-1β at the

[0096] As used herein, the term "antagonist" refers to a binding molecule that inhibits signal transduction activity in the presence of an activating compound.For example, in the case of IL-18, an IL-18 antagonist is a binding molecule that inhibits signal transduction activity in the presence of IL-18 in a human cell assay, such as an IL-18-dependent interferon-gamma (IFN-γ) production assay in human blood cells.An example of an IL-18-dependent IFN-γ production assay in human blood cells is detailed in the following examples.

[0097] The term bivalent bispecific antibody, or bivalent bispecific antibodies (plural), refers to an antibody binding to two different targets, such as IL-18 and IL-1β. Such bivalent bispecific antibodies are also referred to herein as bispecific antibodies.

[0098] Bispecific antibodies are "heterodimers," meaning that one portion is derived from a first antibody specific for a first target and another portion is derived from a second antibody specific for a second target. A "heterodimerizing modification" is a modification to one or both of the antibodies that form the heterodimeric bispecific antibody, or that promotes such formation. An example of a heterodimerization modification of the Fc domains of two IgG1 parts of an antibody intended to form a bispecific is one in which a "knob" with bulky amino acid (aa) side chains (S354C, T366W) in the first heavy chain and a "hole" with small aa side chains (Y349C, T366S, L368A, Y407V) are introduced in the second heavy chain, and an additional disulfide bridge in the CH3 region links both heavy chains (Merchant et al., Nat. Biotechnol., 16:677-681 (1998), page 678, table 1).

[0099] "K D" as used herein refers to the dissociation constant, which is K d and K. a (i.e., K d / K a ) and expressed as molar concentration (M). D Values ​​can be determined using methods well established in the art. D A method for determining is by using surface plasmon resonance, such as a Biacore® system.

[0100] As used herein, the term "affinity" refers to the strength of the interaction between a binding molecule and an antigen at a single antigenic site.

[0101] As used herein, the term "high affinity" for an antibody refers to an antibody that has a KD of 1 nM or less for a target antigen.

[0102] As used herein, the term "subject" includes any human subject who may exhibit symptoms of HS or who may be at risk for HS and who receives a bispecific anti-IL-18 and IL-1β antagonist as described herein.

[0103] As used herein, the term "optimized nucleotide sequence" means that a nucleotide sequence has been modified to encode an amino acid sequence using codons that are preferred in a production cell or organism, typically a eukaryotic cell, such as a Pichia pastoris cell, a Chinese hamster ovary cell (CHO) or a human cell. The optimized nucleotide sequence is modified to completely retain the amino acid sequence originally encoded by the starting nucleotide sequence, also known as the "parent" sequence. The optimized sequences herein have been modified to have codons that are preferred in CHO mammalian cells; however, optimized expression of these sequences in other eukaryotic cells is also contemplated herein.

[0104] The term "identity" refers to the similarity between at least two different sequences. This identity can be expressed as a percentage identity and is determined by standard alignment algorithms, such as the Basic Local Alignment Tool (BLAST) (the algorithm of Altshul et al., (1990) J MoI Biol; 215: 403-410; Needleman et al., (1970) J MoI Biol; 48: 444-453 or the algorithm of Meyers et al., (1988) Comput Appl Biosci; 4: 11-17). The set of parameters can be the Blosum 62 scoring matrix, with a gap penalty of 12, a gap extension penalty of 4, and a frameshift gap penalty of 5. The percent identity between two amino acid or nucleotide sequences can also be determined using the algorithm of E. Meyers and W. Miller (1989) CABIOS;4(1):1-17) as implemented in the ALIGN program (version 2.0), using a PAM120 weight residue table, a gap length of 12, and a gap penalty of 4. Percent identity is usually calculated by comparing sequences of similar length.

[0105] The term "immune response" refers to the action of, for example, lymphocytes, antigen-presenting cells, phagocytes, granulocytes and soluble macromolecules produced by such cells or the liver, including antibodies, cytokines and complement, that result in the selective damage to, destruction of, or elimination from the human body of invading pathogens, pathogen-infected cells or tissues, cancerous cells, or, in the case of autoimmunity or pathological inflammation, normal human cells or tissues.

[0106] "Signal transduction pathway" or "signal transduction activity" refers to a biochemical causal relationship that is generally initiated by a protein-protein interaction, such as the binding of a growth factor to a receptor, resulting in the transmission of a signal from one part of a cell to another part of the cell. Generally, transduction involves the specific phosphorylation of one or more tyrosine, serine, or threonine residues in one or more proteins in the signal transduction that triggers a series of reactions. The penultimate step generally involves a nuclear event that results in a change in gene expression.

[0107] The term "neutralize" and its grammatical variations, as used throughout this application, means to reduce, either completely or partially, the biological activity of a target, optionally in the presence of a binding protein or antibody.

[0108] The term "nucleic acid" or "polynucleotide" refers to deoxyribonucleic acid (DNA) or ribonucleic acid (RNA) and polymers thereof in either single- or double-stranded form. Unless specifically limited, the term encompasses nucleic acids containing known analogs of natural nucleotides that have similar binding properties as the reference nucleic acid and are metabolized in a manner similar to natural nucleotides. Unless otherwise indicated, a particular nucleic acid sequence also implicitly encompasses conservatively modified variants thereof (e.g., degenerate codon substitutions), alleles, orthologs, SNPs, and complementary sequences, as well as the sequence explicitly indicated. Specifically, degenerate codon substitutions can be achieved by generating sequences in which the third position of one or more selected (or all) codons is substituted with mixed-base and / or deoxyinosine residues (Batzer et al., Nucleic Acid Res. 19:5081 (1991); Ohtsuka et al., J. Biol. Chem. 260:2605-2608 (1985); and Rossolini et al., Mol. Cell. Probes 8:91-98 (1994)).

[0109] Nucleotides in a "polynucleotide" or "nucleic acid" may contain modifications, including base modifications such as bromouridine and inosine derivatives, ribose modifications such as phosphorothioate, phosphorodithioate, phosphoroselenoate, phosphorodiselenoate, phosphoroanilothioate, phosphoraniladate, and phosphoroamidate.

[0110] The term "vector" refers to any molecule or entity (e.g., nucleic acid, plasmid, bacteriophage or virus) that is suitable for transformation or transfection of a host cell and contains a nucleic acid sequence that directs and / or regulates (in conjunction with the host cell) the expression of one or more heterologous coding regions to which it is operably linked.

[0111] A "conservative variant" of a sequence encoding a binding molecule, antibody or fragment thereof refers to a sequence containing a conservative amino acid modification. A "conservative amino acid modification" is intended to refer to an amino acid modification that does not significantly affect or alter the binding characteristics of an antibody containing that amino acid sequence. Such conservative modifications include amino acid substitutions, additions and deletions. A conservative amino acid substitution is one in which an amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids with 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, tryptophan), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Modifications can be introduced into the binding proteins of the invention by standard techniques known in the art, such as site-directed mutagenesis and PCR-mediated mutagenesis. Conservative amino acid substitutions can also encompass unnatural amino acid residues that are generally incorporated by chemical peptide synthesis rather than synthesis in a biological system. Unnatural amino acids include, but are not limited to, inverted or peptidomimetic forms of the amino acid moiety.

[0112] The term "epitope" is the portion of an antigen that is recognized by the immune system, such as an antibody or fragment thereof. Within this application, the term "epitope" is used interchangeably for both conformational epitopes and linear epitopes. Conformational epitopes are composed of non-contiguous sections of the amino acid sequence of an antigen, while linear epitopes are formed by a contiguous sequence of amino acids from an antigen.

[0113] A human antibody or fragment thereof may comprise a heavy or light chain variable region or a full-length heavy or light chain that is the "product" of or "derived from" a particular germline sequence if the variable region or full-length chain of the antibody is obtained from a system that uses human germline immunoglobulin genes. Such systems include immunizing transgenic mice carrying human immunoglobulin genes with the antigen of interest or screening a human immunoglobulin gene library displayed on phage with the antigen of interest.

[0114] Human antibodies or fragments thereof that are the "product" or "derived" from human germline immunoglobulin sequences can be identified as such by comparing the amino acid sequence of a human antibody with that of a human germline immunoglobulin and selecting the human germline immunoglobulin sequence that is closest in sequence (i.e., greatest % identity) to the sequence of the human antibody. A human antibody that is the "product" or "derived" from a particular human germline immunoglobulin sequence may contain amino acid differences when compared to the germline sequence, due, for example, to natural somatic mutations or the deliberate introduction of site-specific mutations.

[0115] However, the human antibody selected will generally be at least 90% identical in amino acid sequence to the amino acid sequence encoded by a human germline immunoglobulin gene and will contain amino acid residues that identify the human antibody as human when compared to the germline immunoglobulin amino acid sequence of another species (e.g., mouse germline sequence). In certain cases, a human antibody may be at least 60%, 70%, 80%, 90% or at least 95% or even at least 96%, 97%, 98% or 99% identical in amino acid sequence to the amino acid sequence encoded by a germline immunoglobulin gene. Generally, a human antibody derived from a particular human germline sequence will have no more than 10 amino acid differences from the amino acid sequence encoded by a human germline immunoglobulin gene. In certain cases, a human antibody may exhibit no more than 5 or even no more than 4, 3, 2 or 1 amino acid differences from the amino acid sequence encoded by a germline immunoglobulin gene.

[0116] Human antibodies can be produced by a number of methods known to those skilled in the art. Human antibodies can be produced by the hybridoma method using human myeloma or mouse-human heteromyeloma cell lines (Kozbor, J Immunol; (1984) 133:3001; Brodeur, Monoclonal Isolated Antibody Production Techniques and Applications, pp51-63, Marcel Dekker Inc, 1987). Alternative methods include the use of phage libraries or transgenic mice, both of which utilize human variable region repertoires (Winter G; (1994) Annu Rev Immunol 12:433-455, Green LL, (1999) J Immunol Methods 231:11-23).

[0117] Several lines of transgenic mice are now available in which mouse immunoglobulin loci have been replaced with human immunoglobulin gene segments (Tomizuka K, (2000) Proc Natl Acad Sci, 97:722-727; Fishwild DM (1996) Nature Biotechnol 14:845-851; Mendez MJ, (1997) Nature Genetics 15:146-156). Upon antigen challenge, such mice are capable of producing a repertoire of human antibodies from which antibodies of interest can be selected. Of particular note are the Trimera™ system (Eren R et al, (1988) Immunology 93:154-161), in which human lymphocytes are transplanted into irradiated mice, the Selected Lymphocyte Isolated antibody System (SLAM, Babcook et al, Proc Natl Acad Sci (1996) 93:7843-7848), in which human (or other species) lymphocytes are effectively enrolled through a large number of pooled in vitro isolated antibody generation procedures, followed by deconvolution, limiting dilution and selection procedures, and the Xenomouse™ (Abgenix Inc). An alternative approach uses Morphodoma™ technology and is available from Morphotek Inc.

[0118] Phage display technology can be used to generate human antibodies and fragments thereof (McCafferty; (1990) Nature, 348:552-553 and Griffiths AD et al (1994) EMBO 13:3245-3260). According to this technology, isolated antibody variable domain genes are cloned in frame into either the large or small coat of protein genes of filamentous bacteriophage such as M13 or fd, and displayed (usually with the aid of a helper phage) as functional isolated antibody fragments on the surface of the phage particle. Selection based on the functional properties of the isolated antibodies results in the selection of genes encoding isolated antibodies displaying these properties. Phage display technology can be used to select antigen-specific antibodies from libraries made from human B cells taken from individuals suffering from a disease or disorder or alternatively from non-immunized human donors (Marks; J Mol Bio (1991) 222:581-591). If an intact human isolated antibody is desired to contain an Fc domain, it is necessary to reclone the phage display derived fragment into a mammalian expression vector containing the desired constant region and establish a stable expressing cell line.

[0119] The technique of affinity maturation (Marks; Biotechnol (1992) 10:779-783) can be used to provide improved binding affinity for primary human isolated antibodies by successively replacing heavy and light chain variable regions with natural variants and selecting on the basis of improved binding affinity. Variations of this technique are now available, such as "epitope imprinting" (WO 93 / 06213; Waterhouse; Nucl Acids Res (1993) 21:2265-2266).

[0120] The term "pure", when used in reference to purified bispecific antibodies, relates to the purity and identity of different bispecific antibody combinations and constructs after co-expression in selected cells under conditions in which the cells express the bispecific antibody, and after Protein-A purification using an intact UPLC-MS mass screening approach. Purity or purity refers to the relative quantification of hetero- and homodimeric bbmAbs formed. Using the methods of the invention, correctly formed heterodimeric bbmAb1 can be observed with a relative purity of greater than 85% based on intact mass signal intensity.

[0121] As used herein, the terms "inhibit," "inhibition," or "inhibiting" refer to the alleviation or suppression of a given condition, symptom, or disorder, or disease, or a significant decrease in the baseline activity of a biological activity or process.

[0122] As used herein, the terms "treat", "treating" or "treatment" of any disease or disorder, in one embodiment, refer to ameliorating the disease or disorder (i.e., slowing or arresting or reducing the onset or progression of the disease or at least one of its clinical symptoms). In another embodiment, "treat", "treating" or "treatment" refers to alleviating or ameliorating at least one physical parameter, including those that may not be discernible by the patient. In yet another embodiment, "treat", "treating" or "treatment" refers to modulating the disease or disorder either physically (e.g., stabilization of a discernible symptom), physiologically (e.g., stabilization of a physical parameter), or both. More specifically, the term "treating" the disease HS refers to treating inflammatory lesions in an HS patient (reducing their volume, reducing their size, in number or quality) and / or treating abscesses and inflammatory nodules and / or draining fistulas in an HS patient and / or reducing the amount of scarring and / or reducing the functional limitations associated with scarring. Treating the disease HS also refers to relieving pain, fatigue and / or pruritus associated with HS, reducing purulent discharge and reducing the odor associated with purulent discharge, and / or improving the quality of life and / or reducing work disability for an HS patient.

[0123] As used herein, the term "prevention" refers to delaying the onset or development or progression of a disease or disorder. More specifically, the term "preventing" the disease HS refers to preventing HS plaques and / or the appearance of new lesions; preventing scarring and the functional limitations associated with scarring and / or preventing surgical intervention, particularly for HS.

[0124] As used herein, a subject is "in need of" a treatment if such subject would benefit biologically, medically, or in quality of life from such treatment.

[0125] As used herein, a "therapeutically effective amount" refers to an amount of a bispecific antibody (e.g., bbmAb1) or antigen-binding fragment thereof that simultaneously targets both IL-1β and IL-18 that is effective upon single or multiple administration to a patient (such as a human) to treat, prevent, prevent the onset of, cure, delay, reduce the severity of, ameliorate, or prolong the survival of the patient beyond that expected in the absence of such treatment, of a disorder or at least one symptom of a recurrent disorder. When applied to an individual active ingredient (e.g., bbmAb1) administered alone, the term refers to that ingredient alone. When applied to a combination, the term refers to the combined amounts of the active ingredients that produce the therapeutic effect, whether administered in combination, serially or simultaneously.

[0126] The phrase "therapeutic regimen" refers to a dosing regimen used to treat a disease, e.g., a dosing protocol used during the treatment of HS. A therapeutic regimen can include a loading regimen (or loading dose) followed by a maintenance regimen (or maintenance dose).

[0127] The phrase "loading regimen" or "loading period" refers to a treatment regimen (or a portion of a treatment regimen) used for the initial treatment of a disease. In some embodiments, the disclosed methods, uses, kits, processes and dosing regimens (e.g., methods for treating HS) use a loading regimen (or loading dose). In some cases, the loading period is the period until maximum efficacy is achieved. The general goal of a loading regimen is to provide a high level of drug to the patient during the initial period of the treatment regimen. A loading regimen may involve the physician administering a higher dose of the drug than the dose used during a maintenance regimen, administering the drug more frequently than the physician administers the drug during a maintenance regimen, or both. Dose escalation may occur during or after the loading regimen.

[0128] The phrase "maintenance regimen" or "maintenance period" refers to a treatment regimen (or a portion of a treatment regimen) used for the maintenance of a patient during treatment of a disease, for example to keep the patient in remission for an extended period of time (months or years) after an initial loading regimen or period. In some embodiments, the disclosed methods, uses and dosing regimens use a maintenance dosing regimen. A maintenance dosing regimen may use continuous treatment (e.g., administering a drug at regular intervals, e.g., weekly, biweekly or monthly (every 4 weeks), annually, etc.) or intermittent treatment (e.g., intermittent treatment, intermittent treatment, treatment upon relapse, or treatment upon reaching a certain predetermined criterion (e.g., pain, disease manifestations, etc.). Dose escalation may be performed during a maintenance dosing regimen.

[0129] The phrase "means for administering" is used to refer to any available device for systemically administering a drug to a patient, including, but not limited to, prefilled syringes, vials and syringes, injection pens, autoinjectors, IV drips and bags, pumps, patch pumps, etc. Using such an item, a patient may self-administer a drug (i.e., administer the drug to himself) or a physician may administer the drug.

[0130] 2.IL-18 antibody Particularly preferred IL-18 antibodies or antigen-binding fragments thereof for use in the disclosed methods are human antibodies.

[0131] For ease of reference, the amino acid sequence of the hypervariable region of a specific IL-18 antibody, designated mAb1, and the V L and V H The domains and the complete heavy and light chains are provided in Table 1 below.

[0132] [Table 1]

[0133] In one embodiment, the IL-18 antibody or antigen-binding fragment thereof comprises at least one immunoglobulin heavy chain variable domain (VHV) comprising hypervariable regions CDR1, CDR2 and CDR3, where CDR1 has the amino acid sequence SEQ ID NO:1, CDR2 has the amino acid sequence SEQ ID NO:2 and CDR3 has the amino acid sequence SEQ ID NO:3. H In one embodiment, the IL-18 antibody or antigen-binding fragment thereof comprises at least one immunoglobulin heavy chain variable domain (VHV) comprising hypervariable regions CDR1, CDR2 and CDR3, in which CDR1 has the amino acid sequence SEQ ID NO:4, CDR2 has the amino acid sequence SEQ ID NO:5 and CDR3 has the amino acid sequence SEQ ID NO:6. H ).

[0134] In one embodiment, the IL-18 antibody or antigen-binding fragment thereof comprises at least one immunoglobulin light chain variable domain (VCH) comprising hypervariable regions CDR1, CDR2 and CDR3, in which CDR1 has the amino acid sequence SEQ ID NO:11, CDR2 has the amino acid sequence SEQ ID NO:12 and CDR3 has the amino acid sequence SEQ ID NO:13. L In one embodiment, the IL-18 antibody or antigen-binding fragment thereof comprises at least one immunoglobulin light chain variable domain (VCH) comprising hypervariable regions CDR1, CDR2 and CDR3, in which CDR1 has the amino acid sequence of SEQ ID NO: 14, CDR2 has the amino acid sequence of SEQ ID NO: 15 and CDR3 has the amino acid sequence of SEQ ID NO: 16. L ).

[0135] In one embodiment, the IL-18 antibody or antigen-binding fragment thereof binds at least one immunoglobulin V H domain and at least one immunoglobulin V L a) immunoglobulin V domain; HThe domain comprises (for example in the sequence): i) hypervariable regions CDR1, CDR2 and CDR3, where CDR1 has the amino acid sequence SEQ ID NO:1, CDR2 has the amino acid sequence SEQ ID NO:2 and CDR3 has the amino acid sequence SEQ ID NO:3; or ii) hypervariable regions CDR1, CDR2 and CDR3, where CDR1 has the amino acid sequence SEQ ID NO:4, CDR2 has the amino acid sequence SEQ ID NO:5 and CDR3 has the amino acid sequence SEQ ID NO:6; b) immunoglobulin V L The domain comprises (for example in the sequence): i) hypervariable regions CDR1, CDR2 and CDR3, where CDR1 has the amino acid sequence SEQ ID NO:11, CDR2 has the amino acid sequence SEQ ID NO:12 and CDR3 has the amino acid sequence SEQ ID NO:13 or ii) hypervariable regions CDR1, CDR2 and CDR3, where CDR1 has the amino acid sequence SEQ ID NO:14, CDR2 has the amino acid sequence SEQ ID NO:15 and CDR3 has the amino acid sequence SEQ ID NO:16.

[0136] In one embodiment, the IL-18 antibody or antigen-binding fragment thereof comprises: a) an immunoglobulin heavy chain variable domain (V) comprising the amino acid sequence set forth as SEQ ID NO:7. H ); b) an immunoglobulin light chain variable domain (V) comprising the amino acid sequence set forth as SEQ ID NO: 17 L ); c) an immunoglobulin V comprising the amino acid sequence shown as SEQ ID NO: 7; H Immunoglobulin V comprising the domain and the amino acid sequence shown as SEQ ID NO: 17 L domain; d) immunoglobulin V comprising the hypervariable regions shown as SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3 H domain; e) immunoglobulin V comprising the hypervariable regions shown as SEQ ID NO:11, SEQ ID NO:12 and SEQ ID NO:13 L f) immunoglobulin V domains comprising hypervariable regions shown as SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6 H domain; g) immunoglobulin V comprising the hypervariable regions shown as SEQ ID NO: 14, SEQ ID NO: 15 and SEQ ID NO: 16 L h) immunoglobulin V domains comprising the hypervariable regions shown as SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3 HImmunoglobulin V comprising the domains and hypervariable regions shown as SEQ ID NO:11, SEQ ID NO:12 and SEQ ID NO:13 L Domains; i) immunoglobulin V comprising the hypervariable regions shown as SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6 H Immunoglobulin V comprising the domains and hypervariable regions shown as SEQ ID NO:14, SEQ ID NO:15 and SEQ ID NO:16 L domain; j) a light chain comprising SEQ ID NO:19; k) a heavy chain comprising SEQ ID NO:9; or l) a light chain comprising SEQ ID NO:19 and a heavy chain comprising SEQ ID NO:9.

[0137] In some embodiments, an IL-18 antibody or antigen-binding fragment thereof (e.g., mAb1) comprises the three CDRs of SEQ ID NO: 7. In other embodiments, an IL-18 antibody or antigen-binding fragment thereof comprises the three CDRs of SEQ ID NO: 17. In other embodiments, an IL-18 antibody or antigen-binding fragment thereof comprises the three CDRs of SEQ ID NO: 7 and the three CDRs of SEQ ID NO: 17. In some embodiments, an IL-18 antibody or antigen-binding fragment thereof comprises the three CDRs of SEQ ID NO: 9. In other embodiments, an IL-18 antibody or antigen-binding fragment thereof comprises the three CDRs of SEQ ID NO: 19. In other embodiments, an IL-18 antibody or antigen-binding fragment thereof comprises the three CDRs of SEQ ID NO: 9 and the three CDRs of SEQ ID NO: 19.

[0138] In one embodiment, the IL-18 antibody or antigen-binding fragment thereof (e.g., mAb1) is selected from a human IL-18 antibody comprising at least: a) an immunoglobulin heavy chain or fragment thereof comprising a variable domain comprising hypervariable regions CDR1, CDR2 and CDR3 in sequence, wherein CDR1 has the amino acid sequence SEQ ID NO:1, CDR2 has the amino acid sequence SEQ ID NO:2 and CDR3 has the amino acid sequence SEQ ID NO:3, and a constant portion of a human heavy chain or a fragment thereof; and b) an immunoglobulin light chain or fragment thereof comprising a variable domain comprising hypervariable regions CDR1, CDR2 and CDR3 in sequence, wherein CDR1 has the amino acid sequence SEQ ID NO:11, CDR2 has the amino acid sequence SEQ ID NO:12 and CDR3 has the amino acid sequence SEQ ID NO:13, and a constant portion of a human light chain or a fragment thereof.

[0139] In one embodiment, the IL-18 antibody or antigen-binding fragment thereof (e.g., mAb1) is selected from a human IL-18 antibody comprising at least: a) an immunoglobulin heavy chain or fragment thereof comprising a variable domain comprising hypervariable regions CDR1, CDR2 and CDR3 in sequence, wherein CDR1 has the amino acid sequence SEQ ID NO:4, CDR2 has the amino acid sequence SEQ ID NO:5 and CDR3 has the amino acid sequence SEQ ID NO:6, and a constant portion of a human heavy chain or a fragment thereof; and b) an immunoglobulin light chain or fragment thereof comprising a variable domain comprising hypervariable regions CDR1, CDR2 and CDR3 in sequence, wherein CDR1 has the amino acid sequence SEQ ID NO:14, CDR2 has the amino acid sequence SEQ ID NO:15 and CDR3 has the amino acid sequence SEQ ID NO:16, and a constant portion of a human light chain or a fragment thereof.

[0140] In one embodiment, the IL-18 antibody or antigen-binding fragment thereof is selected from a single chain antibody or antigen-binding fragment thereof comprising an antigen-binding site comprising: a) a first domain comprising hypervariable regions CDR1, CDR2 and CDR3 in sequence, where CDR1 has the amino acid sequence SEQ ID NO:1, CDR2 has the amino acid sequence SEQ ID NO:2 and CDR3 has the amino acid sequence SEQ ID NO:3; b) a second domain comprising hypervariable regions CDR1, CDR2 and CDR3 in sequence, where CDR1 has the amino acid sequence SEQ ID NO:11, CDR2 has the amino acid sequence SEQ ID NO:12 and CDR3 has the amino acid sequence SEQ ID NO:13; and c) a peptide linker binding either to the N-terminus of the first domain and the C-terminus of the second domain or to the C-terminus of the first domain and the N-terminus of the second domain.

[0141] In one embodiment, the IL-18 antibody or antigen-binding fragment thereof (e.g., mAb1) is selected from a single chain antibody or antigen-binding fragment thereof comprising an antigen-binding site comprising: a) a first domain comprising hypervariable regions CDR1, CDR2 and CDR3, in which CDR1 has the amino acid sequence SEQ ID NO:4, CDR2 has the amino acid sequence SEQ ID NO:5 and CDR3 has the amino acid sequence SEQ ID NO:6; b) a second domain comprising hypervariable regions CDR1, CDR2 and CDR3, in which CDR1 has the amino acid sequence SEQ ID NO:14, CDR2 has the amino acid sequence SEQ ID NO:15 and CDR3 has the amino acid sequence SEQ ID NO:16; and c) a peptide linker binding either to the N-terminus of the first domain and the C-terminus of the second domain or to the C-terminus of the first domain and the N-terminus of the second domain.

[0142] V of the IL-18 antibody or antigen-binding fragment thereof used in the disclosed methods H Or V L The domain is represented by SEQ ID NOs: 7 and 17. H Or V L V, which is virtually identical to the domain H and / or V L The human IL-18 antibodies disclosed herein may comprise a heavy chain substantially identical to that set forth as SEQ ID NO:9 and / or a light chain substantially identical to that set forth as SEQ ID NO:19. The human IL-18 antibodies disclosed herein may comprise a heavy chain comprising SEQ ID NO:9 and a light chain comprising SEQ ID NO:19. The human IL-18 antibodies disclosed herein may comprise a) one heavy chain comprising a variable domain having substantially the same amino acid sequence as set forth in SEQ ID NO:7 and a constant portion of a human heavy chain, and b) one light chain comprising a variable domain having substantially the same amino acid sequence as set forth in SEQ ID NO:17 and a constant portion of a human light chain.

[0143] Other preferred IL-18 antagonists (e.g., antibodies) for use in the disclosed methods, kits and treatment regimens are those set forth in U.S. Pat. No. 9,376,489, which is incorporated by reference herein in its entirety.

[0144] 3.IL-1β antibody Particularly preferred IL-1β antibodies or antigen-binding fragments thereof for use in the disclosed methods are human antibodies.

[0145] For ease of reference, the hypervariable region and V of a specific IL-1β antibody, designated mAb2 based on the Kabat and Chothia definitions, are L and V H The domains and complete heavy and light chain amino acid sequences are provided in Table 2 below.

[0146] [Table 2]

[0147] In one embodiment, the IL-1β antibody or antigen-binding fragment thereof comprises at least one immunoglobulin heavy chain variable domain (VHV) comprising hypervariable regions CDR1, CDR2 and CDR3, in which CDR1 has the amino acid sequence SEQ ID NO:21, CDR2 has the amino acid sequence SEQ ID NO:22 and CDR3 has the amino acid sequence SEQ ID NO:23. H In one embodiment, the IL-1β antibody or antigen-binding fragment thereof comprises at least one immunoglobulin heavy chain variable domain (VHV) comprising hypervariable regions CDR1, CDR2 and CDR3, in which CDR1 has the amino acid sequence of SEQ ID NO:24, CDR2 has the amino acid sequence of SEQ ID NO:25 and CDR3 has the amino acid sequence of SEQ ID NO:26. H ).

[0148] In one embodiment, the IL-1β antibody or antigen-binding fragment thereof comprises at least one immunoglobulin light chain variable domain (VCH) comprising hypervariable regions CDR1, CDR2 and CDR3, wherein CDR1 has the amino acid sequence of SEQ ID NO:31, CDR2 has the amino acid sequence of SEQ ID NO:32 and CDR3 has the amino acid sequence of SEQ ID NO:33. LIn one embodiment, the IL-1β antibody or antigen-binding fragment thereof comprises at least one immunoglobulin light chain variable domain (VCH) comprising hypervariable regions CDR1, CDR2 and CDR3, in which CDR1 has the amino acid sequence of SEQ ID NO:34, CDR2 has the amino acid sequence of SEQ ID NO:35 and CDR3 has the amino acid sequence of SEQ ID NO:36. L ).

[0149] In one embodiment, the IL-1β antibody or antigen-binding fragment thereof binds to at least one immunoglobulin V H domain and at least one immunoglobulin V L a) immunoglobulin V domain; H The domain comprises (for example in sequence): i) hypervariable regions CDR1, CDR2 and CDR3, where CDR1 has the amino acid sequence SEQ ID NO:21, CDR2 has the amino acid sequence SEQ ID NO:22 and CDR3 has the amino acid sequence SEQ ID NO:23; or ii) hypervariable regions CDR1, CDR2 and CDR3, where CDR1 has the amino acid sequence SEQ ID NO:24, CDR2 has the amino acid sequence SEQ ID NO:25 and CDR3 has the amino acid sequence SEQ ID NO:26; b) immunoglobulin V L The domain comprises (for example in sequence): i) hypervariable regions CDR1, CDR2 and CDR3, where CDR1 has the amino acid sequence SEQ ID NO:31, CDR2 has the amino acid sequence SEQ ID NO:32 and CDR3 has the amino acid sequence SEQ ID NO:33 or ii) hypervariable regions CDR1, CDR2 and CDR3, where CDR1 has the amino acid sequence SEQ ID NO:34, CDR2 has the amino acid sequence SEQ ID NO:35 and CDR3 has the amino acid sequence SEQ ID NO:36.

[0150] In one embodiment, the IL-1β antibody, or antigen-binding fragment thereof, comprises: a) an immunoglobulin heavy chain variable domain (V) comprising the amino acid sequence set forth as SEQ ID NO:27. H ); b) an immunoglobulin light chain variable domain (V) comprising the amino acid sequence set forth as SEQ ID NO: 37 L ); c) an immunoglobulin V comprising the amino acid sequence shown as SEQ ID NO: 27 HImmunoglobulin V comprising the domain and the amino acid sequence shown as SEQ ID NO:37 L domain; d) immunoglobulin V comprising the hypervariable regions shown as SEQ ID NO:21, SEQ ID NO:22 and SEQ ID NO:23 H domain; e) immunoglobulin V comprising the hypervariable regions shown as SEQ ID NO: 31, SEQ ID NO: 32 and SEQ ID NO: 33 L domain; f) immunoglobulin V comprising the hypervariable regions shown as SEQ ID NO:24, SEQ ID NO:25 and SEQ ID NO:26 H domain; g) immunoglobulin V comprising the hypervariable regions shown as SEQ ID NO: 34, SEQ ID NO: 35 and SEQ ID NO: 36 L domain; h) immunoglobulin V comprising the hypervariable regions shown as SEQ ID NO:21, SEQ ID NO:22 and SEQ ID NO:23 H Immunoglobulin V comprising the domains and hypervariable regions shown as SEQ ID NO:31, SEQ ID NO:32 and SEQ ID NO:33 L Domains; i) immunoglobulin V comprising the hypervariable regions shown as SEQ ID NO:24, SEQ ID NO:25 and SEQ ID NO:26 H Immunoglobulin V comprising the domains and hypervariable regions shown as SEQ ID NO:34, SEQ ID NO:35 and SEQ ID NO:36 L domain; j) a light chain comprising SEQ ID NO:37; k) a heavy chain comprising SEQ ID NO:29; or l) a light chain comprising SEQ ID NO:39 and a heavy chain comprising SEQ ID NO:29.

[0151] In some embodiments, an IL-1β antibody or antigen-binding fragment thereof (e.g., mAb2) comprises the three CDRs of SEQ ID NO: 37. In other embodiments, an IL-1β antibody or antigen-binding fragment thereof comprises the three CDRs of SEQ ID NO: 27. In other embodiments, an IL-1β antibody or antigen-binding fragment thereof comprises the three CDRs of SEQ ID NO: 37 and the three CDRs of SEQ ID NO: 27. In some embodiments, an IL-1β antibody or antigen-binding fragment thereof comprises the three CDRs of SEQ ID NO: 39. In other embodiments, an IL-1β antibody or antigen-binding fragment thereof comprises the three CDRs of SEQ ID NO: 29. In other embodiments, an IL-1β antibody or antigen-binding fragment thereof comprises the three CDRs of SEQ ID NO: 39 and the three CDRs of SEQ ID NO: 29.

[0152] In one embodiment, the IL-1β antibody or antigen-binding fragment thereof (e.g., mAb2) is selected from a human IL-1β antibody comprising at least: a) an immunoglobulin heavy chain or a fragment thereof comprising a variable domain comprising hypervariable regions CDR1, CDR2 and CDR3 in their sequence, wherein CDR1 has the amino acid sequence SEQ ID NO:21, CDR2 has the amino acid sequence SEQ ID NO:22 and CDR3 has the amino acid sequence SEQ ID NO:23, and a constant portion of a human heavy chain or a fragment thereof; and b) an immunoglobulin light chain or a fragment thereof comprising a variable domain comprising hypervariable regions CDR1, CDR2 and CDR3 in their sequence, wherein CDR1 has the amino acid sequence SEQ ID NO:31, CDR2 has the amino acid sequence SEQ ID NO:32 and CDR3 has the amino acid sequence SEQ ID NO:33, and a constant portion of a human light chain or a fragment thereof.

[0153] In one embodiment, the IL-1β antibody or antigen-binding fragment thereof (e.g., mAb2) is selected from a human IL-1β antibody comprising at least: a) an immunoglobulin heavy chain or fragment thereof comprising a variable domain comprising hypervariable regions CDR1, CDR2 and CDR3 in sequence, wherein CDR1 has the amino acid sequence of SEQ ID NO:24, CDR2 has the amino acid sequence of SEQ ID NO:25, and CDR3 has the amino acid sequence of SEQ ID NO:26, and a constant portion of a human heavy chain or a fragment thereof; and b) an immunoglobulin light chain or fragment thereof comprising a variable domain comprising hypervariable regions CDR1, CDR2 and CDR3 in sequence, wherein CDR1 has the amino acid sequence of SEQ ID NO:34, CDR2 has the amino acid sequence of SEQ ID NO:35, and CDR3 has the amino acid sequence of SEQ ID NO:36, and a constant portion of a human light chain or a fragment thereof.

[0154] In one embodiment, the IL-1β antibody or antigen-binding fragment thereof is selected from a single chain antibody or antigen-binding fragment thereof comprising an antigen-binding site comprising: a) a first domain comprising hypervariable regions CDR1, CDR2 and CDR3, in which CDR1 has the amino acid sequence SEQ ID NO:21, CDR2 has the amino acid sequence SEQ ID NO:22 and CDR3 has the amino acid sequence SEQ ID NO:23; b) a second domain comprising hypervariable regions CDR1, CDR2 and CDR3, in which CDR1 has the amino acid sequence SEQ ID NO:31, CDR2 has the amino acid sequence SEQ ID NO:32 and CDR3 has the amino acid sequence SEQ ID NO:33; and c) a peptide linker binding either to the N-terminus of the first domain and the C-terminus of the second domain or to the C-terminus of the first domain and the N-terminus of the second domain.

[0155] In one embodiment, the IL-1β antibody or antigen-binding fragment thereof (e.g., mAb2) is selected from single chain antibodies or antigen-binding fragments thereof, comprising an antigen-binding site comprising: a) a first domain comprising hypervariable regions CDR1, CDR2 and CDR3, in which CDR1 has the amino acid sequence SEQ ID NO:24, CDR2 has the amino acid sequence SEQ ID NO:25 and CDR3 has the amino acid sequence SEQ ID NO:26; b) a second domain comprising hypervariable regions CDR1, CDR2 and CDR3, in which CDR1 has the amino acid sequence SEQ ID NO:34, CDR2 has the amino acid sequence SEQ ID NO:35 and CDR3 has the amino acid sequence SEQ ID NO:36; and c) a peptide linker binding either to the N-terminus of the first domain and the C-terminus of the second domain or to the C-terminus of the first domain and the N-terminus of the second domain.

[0156] V of the IL-1β antibody used in the disclosed methods H Or V L The domain or antigen-binding fragment thereof is H Or V L V, which is virtually identical to the domain H and / or V LThe human IL-1β antibodies disclosed herein may comprise a heavy chain substantially identical to that set forth as SEQ ID NO:29 and / or a light chain substantially identical to that set forth as SEQ ID NO:39. The human IL-1β antibodies disclosed herein may comprise a heavy chain comprising SEQ ID NO:29 and a light chain comprising SEQ ID NO:39. The human IL-1β antibodies disclosed herein may comprise: a) one heavy chain comprising a variable domain having substantially the same amino acid sequence as set forth in SEQ ID NO:27 and a constant portion of a human heavy chain; and b) one light chain comprising a variable domain having substantially the same amino acid sequence as set forth in SEQ ID NO:37 and a constant portion of a human light chain.

[0157] Other preferred IL-1β antagonists (e.g., antibodies) for use in the disclosed methods, kits, and treatment regimens are those set forth in U.S. Pat. Nos. 7,446,175 or 7,993,878 or 8,273,350, which are incorporated by reference in their entireties herein.

[0158] 4.Fc modification In addition to, or instead of, modifications made within the framework or CDR regions, the antibodies of the invention may be engineered to include modifications within the Fc region, generally to alter one or more functional properties of the antibody, such as serum half-life, complement binding, Fc receptor binding, and / or antigen-dependent cellular cytotoxicity. Further, an antibody of the invention may be chemically modified (e.g., one or more chemical moieties may be linked to the antibody) or its glycosylation may be altered, again to alter one or more functional properties of the antibody. Each of these embodiments is described in further detail below. The numbering of residues in the Fc region is that of the EU numbering scheme of Edelman et al., PNAS, 1969 May, 63(1):78-85.

[0159] In one embodiment, the hinge region of CH1 is modified such that the number of cysteine ​​residues in the hinge region is altered, e.g., increased or decreased. This approach is further described in U.S. Patent No. 5,677,425 by Bodmer et al. The number of cysteine ​​residues in the hinge region of CH1 is altered, e.g., to facilitate assembly of the light and heavy chains or to increase or decrease the stability of the antibody.

[0160] In another embodiment, mutations are made to the Fc hinge region of antibody to reduce the biological half-life of the antibody.More specifically, one or more amino acid mutations are introduced into the CH2-CH3 domain interface region of Fc-hinge fragment, so that the antibody is impaired in Staphylococcus protein A (SpA) binding compared to native Fc-hinge domain SpA binding.This approach is described in more detail in US Patent No. 6,165,745 by Ward et al.

[0161] In another embodiment, the antibody is modified to increase its biological half-life. Various approaches are possible. For example, one or more of the following mutations can be introduced: T252L, T254S, T256F, as described in US Pat. No. 6,277,375 to Ward. Alternatively, to increase biological half-life, the antibody can be modified in the CH1 or CL region to contain a salvage receptor binding epitope taken from two loops of the CH2 domain of the Fc region of IgG, as described in US Pat. Nos. 5,869,046 and 6,121,022 by Presta et al.

[0162] In yet another embodiment, the Fc region is modified by replacing at least one amino acid residue with a different amino acid residue to alter the effector function of the antibody. For example, one or more amino acids may be replaced with a different amino acid residue such that the affinity of the antibody to an effector ligand is altered but retains the antigen-binding ability of the parent antibody. The effector ligand to which the affinity is altered may be, for example, an Fc receptor or the C1 component of complement. This approach is described in further detail in U.S. Patent Nos. 5,624,821 and 5,648,260, both by Winter et al.

[0163] In another embodiment, one or more selected amino acid residues may be replaced with a different amino acid residue such that the antibody has altered C1q binding and / or reduced or eliminated complement dependent cytotoxicity (CDC). This approach is described in further detail in U.S. Patent No. 6,194,551 by Idusogie et al.

[0164] In another embodiment, one or more amino acid residues are altered to thereby alter the ability of the antibody to fix complement. This approach is further described in WO 94 / 29351 by Bodmer et al.

[0165] In yet another embodiment, the Fc region is modified to improve the ability of the antibody to mediate antibody-dependent cellular cytotoxicity and / or to improve the affinity of the antibody for the Fcγ receptor by modifying one or more amino acids. This approach is described further in WO 00 / 42072 by Presta. Additionally, the binding sites on human IgG1 for FcγRI, FcγRII, FcγRIII and FcRn have been mapped and mutants with improved binding have been described (see Shields, RL et al, (2001) J Biol Chem 276:6591-6604).

[0166] In certain embodiments, an Fc domain of IgG1 isotype is used. In some specific embodiments, mutants of IgG1 Fc fragments are used, such as silent IgG1 Fc, which reduce or eliminate the ability of the fusion polypeptide to mediate antibody-dependent cellular cytotoxicity (ADCC) and / or bind to Fcγ receptors. An example of an IgG1 isotype silent mutant in which leucine residues at amino acid positions 234 and 235 are replaced by alanine residues is described in Hezareh et al, J. Virol (2001); 75(24): 12161-8.

[0167] In certain embodiments, the Fc domain is mutant to prevent glycosylation at position 297 of the Fc domain. For example, the Fc domain contains an amino acid substitution of an asparagine residue at position 297. Examples of such amino acid substitutions are replacement of N297 with glycine or alanine.

[0168] Mutations in the Fc region of antibodies can result in suppression of effector functions and have been described in the art: LALA and N297A (Strohl, W., 2009, Curr. Opin. Biotechnol. vol. 20(6): 685-691); and D265A (Baudino et al., 2008, J. Immunol. 181: 6664-69; Strohl, W. supra); and DAPA (D265A and P329A) (Shields RL., J Biol Chem. 2001; 276(9): 6591-604; US Patent Publication No. 2015 / 0320880). Examples of silent Fc IgG1 antibodies include LALA mutants that contain the so-called L234A and L235A mutations in the IgG1 Fc amino acid sequence. Another example of a silent IgG1 antibody contains the D265A mutation. Another example of a silent IgG1 antibody is the so-called DAPA mutant, which contains D265A and P329A mutations relative to the IgG1 Fc amino acid sequence. Another silent IgG1 antibody contains a N297A mutation, resulting in an aglycosylated / aglycosylated antibody. Further Fc mutations to produce effector function suppression are described in WO2014 / 145806 (e.g., FIG. 7 of WO2014 / 145806), which is incorporated by reference in its entirety. One example from WO2014 / 145806 of a silent IgG1 antibody contains E233P, L234V, L235A and S267K mutations and a deletion of G236 (G236del). Another example of a silent IgG1 antibody from WO 2014 / 145806 includes the E233P, L234V and L235A mutations and a deletion of G236 (G236del). Another example of a silent IgG1 antibody from WO 2014 / 145806 includes the S267K mutation.

[0169] In yet another embodiment, the glycosylation of the antibody is modified. For example, an aglycosylated antibody (i.e., the antibody lacks glycosylation) can be generated. The glycosylation can be altered to, for example, improve the affinity of the antibody for an antigen. Such carbohydrate modifications can be accomplished, for example, by altering one or more sites of glycosylation within the antibody sequence. For example, one or more variable region framework glycosylation sites can be eliminated, thereby making one or more amino acid substitutions that eliminate glycosylation at that site. Such aglycosylation can improve the affinity of the antibody for an antigen. Such approaches are described in further detail in U.S. Pat. Nos. 5,714,350 and 6,350,861 by Co et al.

[0170] Additionally or alternatively, antibodies may be made with altered types of glycosylation, such as hypofucosylated antibodies with reduced amounts of fucosyl residues or antibodies with increased bisection of GlcNac structures. Such altered glycosylation patterns have been shown to improve the ADCC ability of antibodies. Such carbohydrate modifications may be accomplished, for example, by expressing the antibody in a host cell with altered glycosylation machinery. Cells with altered glycosylation machinery have been described in the art and may be used as host cells to express the recombinant antibodies of the invention, thereby producing antibodies with altered glycosylation. For example, EP 1,176,195 by Hang et al. describes cell lines with a functional disruption of the FUT8 gene, which encodes a fucosyltransferase, such that antibodies expressed in such cell lines exhibit hypofucosylation. Thus, in one embodiment, the antibodies of the invention are produced by recombinant expression in a cell line that exhibits a hypofucosylation pattern, such as a mammalian cell line in which expression of the FUT8 gene encoding a fucosyltransferase is insufficient. WO 03 / 035835 by Presta describes a mutant CHO cell line, Lecl3 cells, that has a reduced ability to attach fucose to Asn(297)-linked carbohydrates, which also results in hypofucosylation of antibodies expressed in the host cells (see also Shields, RR et al., 2002 J. Biol. Chem. 277:26733-26740). WO 99 / 54342 by Umana et al. describes cell lines engineered to express glycoprotein-modifying glycosyltransferases, such as beta(1,4)-N-acetylglucosaminyltransferase III (GnTIII), such that antibodies expressed in the engineered cell lines exhibit an increase in bisecting GlcNac structures that increase the ADCC activity of the antibodies (see also Umana et al., 1999 Nat. Biotech. 17:176-180).Alternatively, the antibodies of the invention can be produced in yeast or filamentous fungi that have been engineered for a mammalian-like glycosylation pattern and are capable of producing antibodies that lack fucose as a glycosylation pattern (see, for example, EP 1297172 B1).

[0171] Another modification of the antibodies herein contemplated by the present invention is pegylation. For example, the antibody may be pegylated to extend the biological (e.g. serum) half-life of the antibody. To pegylate an antibody, the antibody or a fragment thereof is generally reacted with polyethylene glycol (PEG), such as a reactive ester or aldehyde derivative of PEG, under conditions such that one or more PEG groups become linked to the antibody or antibody fragment. PEGylation may be performed with a reactive PEG molecule (or an analogous reactive water-soluble polymer) by acylation or alkylation reactions. As used herein, the term "polyethylene glycol" is intended to encompass any of the forms of PEG that have been used to derivatize other proteins, such as mono (C1-C10) alkoxy- or aryloxy-polyethylene glycol or polyethylene glycol-maleimide. In certain embodiments, the antibody to be pegylated is an aglycosylated antibody. Methods for pegylation of proteins are known in the art and may be applied to the antibodies of the present invention. See, for example, EP 0154316 to Nishimura et al. and EP 0401384 to Ishikawa et al.

[0172] Another modification of the antibodies contemplated by the present invention is the conjugation or protein fusion of at least the antigen-binding region of the antibody of the invention to a serum protein, such as human serum albumin or a fragment thereof, in order to increase the half-life of the resulting molecule. Such an approach is described, for example, in Ballance et al., EP 0 322 094.

[0173] Another modification of the antibody contemplated by the present invention is one or more modifications to increase the formation of heterodimeric bispecific antibodies.Various approaches available in the art can be used to promote the dimerization of the two heavy chain domains of a bispecific antibody, such as a bbmAb, as disclosed, for example, in EP1870459A1; U.S. Pat. No. 5,582,996; U.S. Pat. No. 5,731,168; U.S. Pat. No. 5,910,573; U.S. Pat. No. 5,932,448; U.S. Pat. No. 6,833,441; U.S. Pat. No. 7,183,076; U.S. Patent Application Publication No. 2006204493A1; and WO2009 / 089004A1, the contents of which are incorporated herein in their entirety.

[0174] The generation of bispecific antibodies using knobs-into-holes is disclosed, for example, in WO 1996 / 027011, Ridgway et al., (1996), and Merchant et al. (1998).

[0175] In some implementations of the disclosed methods of treatment or use, a therapeutically effective amount of a bispecific antibody that simultaneously targets both IL-1β and IL-18, such as bbmAb1, must be administered to a subject in need thereof. It is understood that variations in dosing schedule may be appropriate for certain patients. Thus, dosing (e.g., of bbmAb1) may be more frequent, such as daily, biweekly, or weekly.

[0176] Some patients may benefit from a loading dosing regimen (e.g., daily dosing for several days / [e.g., dosing for 1-4 days, e.g., on days 0, 1, 2 and / or 3], where bbmAb1 may be administered weekly, biweekly or every 4 weeks for several weeks, followed by a maintenance dosing regimen beginning, e.g., on week 3 or 4. In some embodiments, the duration of administration of a bispecific antibody that simultaneously targets both IL-1β and IL-18, e.g., bbmAb1, may be 1 day, 2 days, 3 days, In some embodiments, the period of administration of a bispecific antibody that simultaneously targets both IL-1β and IL-18, e.g., bbmAb1, is 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months or more.

[0177] It will be appreciated that dose escalation may be appropriate for certain patients, e.g., patients based on disease severity, e.g., patients who show an inadequate response to treatment with bbmAb1. Thus, the dosage (intravenous (iv)) may be greater than about 10 mg / kg, e.g., about 11 mg / kg, 12 mg / kg, 15 mg / kg, 20 mg / kg, 25 mg / kg, 30 mg / kg, 35 mg / kg, etc. Additionally, the subcutaneous (sc) dosage (loading or maintenance dose) may be greater than about 50 mg to about 900 mg sc, e.g., about 75 mg, about 100 mg, about 125 mg, about 175 mg, about 200 mg, about 250 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 600 mg, etc.

[0178] It will also be appreciated that dose reductions may be appropriate for certain patients, such as those who experience adverse events or reactions to treatment with bbmAb1. Thus, the dosage may be less than about 10 mg / kg, e.g., about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, or about 9 mg / kg. In some embodiments, the bbmAB1 dose may be adjusted as determined by a physician.

[0179] In some embodiments, the bbmAB1 antibody may be administered to a patient as a single dose of 10 mg / kg delivered iv, which dose may be adjusted to a higher or lower dose, e.g., about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, about 4 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg or about 9 mg / kg, or for example about 11 mg / kg, 12 mg / kg, 15 mg / kg, 20 mg / kg, 25 mg / kg, 30 mg / kg, 35 mg / kg, etc., if necessary, as determined by a physician.

[0180] In some embodiments, the bbmAB1 antibody may be administered to a patient at an initial dose of 10 mg / kg delivered iv, and then the dose may be adjusted to a higher or lower dose as needed, as determined by the physician.

[0181] In a specific embodiment, 10 mg / kg bbmAB1 is administered on day 1.

[0182] In a specific embodiment, 10 mg / kg bbmAB1 is administered on day 1 (D1) and on days 2 (D2), D3, D4, D5, D6, D7, D8, D9, D10, D11, D12, D13 and / or D14.

[0183] In another specific embodiment, 10 mg / kg bbmAB1 is administered iv on day 1. EXAMPLES

[0184] Example 1: Multiple inflammasomes appear to be involved in HS. Analysis of HS transcriptomes from patients shows elevated mRNA levels of AIM2, NLRC4, NLRP7 and NLRP3 inflammasomes in HS lesions compared to levels in non-lesional or healthy tissues (Figure 1). The expression levels of all these inflammasome genes were obviously higher in HS lesion samples than in HS non-lesional and healthy samples, indicating that multiple inflammasomes are likely involved in HS pathophysiology.

[0185] Example 2: Both IL-1β and IL-18 signaling pathways are active in HS. (a) Transcriptomics of skin biopsies Transcriptomic analysis of 18 lesional HS biopsies, 6 perilesional biopsies, and 7 nonlesional biopsies versus 8 biopsies from healthy skin donors was obtained as follows: Homogenates were prepared from flash-frozen skin tissues using the recommended buffer from the Qiagen RNeasy mini kit. Cellular total RNA was extracted according to the manufacturer's protocol. cDNA was prepared for the samples from the same starting amount of RNA using the High Capacity cDNA Reverse Transcription Kit (Applied Biosystems). Samples were processed by CiToxLAB France on Affymetrix HG_U133_Plus2 microarrays. RMA-normalized data were analyzed using GeneSpring 11.5.1 (Agilent Technologies, Santa Clara, CA). Data were first subjected to standard QC controls by CiToxLAB and in GeneSpring (PCA, hybridization controls). This was then filtered by expression level for probe sets that were above the 20th percentile in 100% of the samples in any one of the conditions before further analysis. The dataset was deposited in the NCBI GEO database (GSE148027). The results were visualized using TIBCO Sporfire Analyst.

[0186] (b) Transcriptomics of cytokine-stimulated PBMCs (from healthy donors). PBMCs were stimulated with recombinant cytokines at 7x106 PBMCs / well (in 12-well plates) in a final volume of 1.5mL in RPMI medium. Recombinant cytokines were added at the following final concentrations: 10ng / mL recombinant IL-1β, 3nM recombinant IL-18 and 1ng / mL recombinant IL-12. Cells were harvested 6 hours after stimulation in cell culture at 37°C and 5% CO2.

[0187] For RNA isolation, cells were pelleted and the pellet was dissolved in 350 μL of Qiagen RTL buffer with 2% β-mercaptoethanol and frozen at -20 °C or -80 °C until all test samples were collected. RNA isolation was performed using Qiagen standard protocols. The different RNA samples were processed by CiToxLAB France on Affymetrix HG_U133_Plus2 microarrays as described above. Entities (probe sets) with at least 100% of the samples having values ​​above the 20th percentile in any one of the experimental conditions were kept. Differentially expressed genes (DEGs) were identified using the "Filter on Volcano Plot" property in GeneSpring. Probe sets with corrected p-values ​​below 0.05 and fold changes above 2.0 were considered as differentially expressed using filtered genes (20.0-100.0th percentile expression) together with an unpaired T-test. Benjamini-Hochberg multiple testing correction was used when possible.

[0188] The resulting gene list was used as a "cytokine signaling signature" and the expression levels of the defined signature were examined in the HS transcriptome dataset without using any associated values ​​from the original experiments.

[0189] (c) Creation of spotfire transcriptome heatmap For every single skin biopsy, the average expression levels of differentially upregulated genes from cytokine-stimulated PBMCs were calculated, and color coding with increasing color intensity was due to increased average expression levels of individual PBMC signatures (Figure 2).

[0190] We provide evidence that both IL-1b and IL-18 signaling are present and active in the lesional skin of HS patients and that these two cytokines likely play a role in HS pathophysiology.

[0191] Example 3: The production of bbmAb1 is described in detail in Examples 1-5 of patent application WO 2018 / 229612. Example 1 of WO 2018 / 229612 is incorporated herein by reference in its entirety, including (1) vector construction, (2) host cell lines and gene transfer, (3) cell selection and sorting, (4) cell growth, (5) clonal stability, (6) production, (7) analytical characterization and purity assessment, and (8) analytical results.

[0192] The resulting bbmAb1 is a bispecific IgG1 with LALA silencing mutations that simultaneously binds two distinct targets, IL-1β and IL-18. The antibody combines two distinct antigen-binding arms (Fab fragments), while the Fab for IL-1β is based on mAb2 and contains a kappa light chain (Vk6). The Fab for IL-18 is based on mAb1 and is composed of a lambda light chain (Vλ1). To drive heterodimerization of the Fc domains during expression, a "knob" with bulky amino acid (aa) side chains (S354C and T366W) in the mAb1 heavy chain and a "hole" with small aa side chains (Y349C, T366S, L368A, Y407V) were introduced in the mAb2 heavy chain.

[0193] For ease of reference, the hypervariable regions and V domains of bbmAb1 according to the Kabat and Chothia definitions are L and V HThe domains and complete heavy and light chain amino acid sequences are provided in Table 3 below.

[0194] [Table 3]

[0195] [Table 4]

[0196] In one embodiment, the IL-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS comprises a first immunoglobulin heavy chain variable domain (VCH) comprising hypervariable regions CDR1, CDR2 and CDR3. H1 ), wherein said CDR1 has the amino acid sequence SEQ ID NO: 76, said CDR2 has the amino acid sequence SEQ ID NO: 77 and said CDR3 has the amino acid sequence SEQ ID NO: 78. In one embodiment, the IL-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS comprises a first immunoglobulin heavy chain variable domain (VHV) comprising hypervariable regions CDR1, CDR2 and CDR3. H1 ), wherein said CDR1 has the amino acid sequence SEQ ID NO: 79, said CDR2 has the amino acid sequence SEQ ID NO: 80 and said CDR3 has the amino acid sequence SEQ ID NO: 81. In one embodiment, (i) the IL-18 / IL-1β bispecific antibody for use in the disclosed treatment or prevention of HS comprises a first immunoglobulin heavy chain variable domain (VHV) comprising hypervariable regions CDR1, CDR2 and CDR3. H1 ), wherein said CDR1 has the amino acid sequence SEQ ID NO:82, said CDR2 has the amino acid sequence SEQ ID NO:83, and said CDR3 has the amino acid sequence SEQ ID NO:84.

[0197] In one embodiment, the IL-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS comprises a second immunoglobulin heavy chain variable domain (VHc) comprising hypervariable regions CDR1, CDR2 and CDR3, wherein CDR1 has the amino acid sequence SEQ ID NO: 44, CDR2 has the amino acid sequence SEQ ID NO: 45 and CDR3 has the amino acid sequence SEQ ID NO: 46.H2 In one embodiment, the IL-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS comprises a second immunoglobulin heavy chain variable domain (VHc) comprising hypervariable regions CDR1, CDR2 and CDR3, wherein CDR1 has the amino acid sequence SEQ ID NO: 47, CDR2 has the amino acid sequence SEQ ID NO: 48 and CDR3 has the amino acid sequence SEQ ID NO: 49. H2 In one embodiment, the IL-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS comprises a second immunoglobulin heavy chain variable domain (VHc) comprising hypervariable regions CDR1, CDR2 and CDR3, wherein CDR1 has the amino acid sequence SEQ ID NO: 50, CDR2 has the amino acid sequence SEQ ID NO: 51 and CDR3 has the amino acid sequence SEQ ID NO: 52. H2 ).

[0198] In one embodiment, the IL-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS comprises a first immunoglobulin light chain variable domain (VCH) comprising hypervariable regions CDR1, CDR2 and CDR3, wherein CDR1 has the amino acid sequence SEQ ID NO: 92, CDR2 has the amino acid sequence SEQ ID NO: 93 and CDR3 has the amino acid sequence SEQ ID NO: 94. L1 In one embodiment, the IL-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS comprises a first immunoglobulin light chain variable domain (VCH) comprising hypervariable regions CDR1, CDR2 and CDR3, wherein CDR1 has the amino acid sequence SEQ ID NO: 95, CDR2 has the amino acid sequence SEQ ID NO: 96 and CDR3 has the amino acid sequence SEQ ID NO: 97. L1 In one embodiment, the IL-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS comprises a first immunoglobulin light chain variable domain (V) comprising hypervariable regions CDR1, CDR2 and CDR3, in which CDR1 has the amino acid sequence SEQ ID NO: 98, CDR2 has the amino acid sequence SEQ ID NO: 99 and CDR3 has the amino acid sequence SEQ ID NO: 100. L1 ).

[0199] In one embodiment, the IL-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS comprises a second immunoglobulin light chain variable domain (VCH) comprising hypervariable regions CDR1, CDR2 and CDR3, wherein CDR1 has the amino acid sequence SEQ ID NO: 60, CDR2 has the amino acid sequence SEQ ID NO: 61 and CDR3 has the amino acid sequence SEQ ID NO: 62. L2 In one embodiment, the IL-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS comprises a second immunoglobulin light chain variable domain (V) comprising hypervariable regions CDR1, CDR2 and CDR3, in which CDR1 has the amino acid sequence SEQ ID NO: 63, CDR2 has the amino acid sequence SEQ ID NO: 64 and CDR3 has the amino acid sequence SEQ ID NO: 65. L2 In one embodiment, the IL-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS comprises a second immunoglobulin light chain variable domain (V) comprising hypervariable regions CDR1, CDR2 and CDR3, in which CDR1 has the amino acid sequence SEQ ID NO: 66, CDR2 has the amino acid sequence SEQ ID NO: 67 and CDR3 has the amino acid sequence SEQ ID NO: 68. L2 ).

[0200] In one embodiment, the IL-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS comprises a first immunoglobulin V H1 Domain and first immunoglobulin V L1 a) a first immunoglobulin V domain; H1The domain comprises (for example in the sequence): i) hypervariable regions CDR1, CDR2 and CDR3, where CDR1 has the amino acid sequence SEQ ID NO: 76, CDR2 has the amino acid sequence SEQ ID NO: 77 and CDR3 has the amino acid sequence SEQ ID NO: 78; or ii) hypervariable regions CDR1, CDR2 and CDR3, where CDR1 has the amino acid sequence SEQ ID NO: 79, CDR2 has the amino acid sequence SEQ ID NO: 80 and CDR3 has the amino acid sequence SEQ ID NO: 81; or iii) hypervariable regions CDR1, CDR2 and CDR3, where CDR1 has the amino acid sequence SEQ ID NO: 82, CDR2 has the amino acid sequence SEQ ID NO: 83 and CDR3 has the amino acid sequence SEQ ID NO: 84; and b) a first immunoglobulin V L1 The domain comprises (for example in sequence): i) hypervariable regions CDR1, CDR2 and CDR3, where CDR1 has the amino acid sequence SEQ ID NO: 92, CDR2 has the amino acid sequence SEQ ID NO: 93 and CDR3 has the amino acid sequence SEQ ID NO: 94; or ii) hypervariable regions CDR1, CDR2 and CDR3, where CDR1 has the amino acid sequence SEQ ID NO: 95, CDR2 has the amino acid sequence SEQ ID NO: 96 and CDR3 has the amino acid sequence SEQ ID NO: 97; or iii) hypervariable regions CDR1, CDR2 and CDR3, where CDR1 has the amino acid sequence SEQ ID NO: 98, CDR2 has the amino acid sequence SEQ ID NO: 99 and CDR3 has the amino acid sequence SEQ ID NO: 100.

[0201] In one embodiment, the IL-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS comprises a second immunoglobulin V H2 Domain and second immunoglobulin V L2 domain, wherein: a) a second immunoglobulin V H2The domain comprises (for example in sequence): i) hypervariable regions CDR1, CDR2 and CDR3, where CDR1 has the amino acid sequence SEQ ID NO: 44, CDR2 has the amino acid sequence SEQ ID NO: 45 and CDR3 has the amino acid sequence SEQ ID NO: 46; or ii) hypervariable regions CDR1, CDR2 and CDR3, where CDR1 has the amino acid sequence SEQ ID NO: 47, CDR2 has the amino acid sequence SEQ ID NO: 48 and CDR3 has the amino acid sequence SEQ ID NO: 49; or iii) hypervariable regions CDR1, CDR2 and CDR3, where CDR1 has the amino acid sequence SEQ ID NO: 50, CDR2 has the amino acid sequence SEQ ID NO: 51 and CDR3 has the amino acid sequence SEQ ID NO: 52; and b) a second immunoglobulin V L2 The domain comprises (for example in sequence): i) hypervariable regions CDR1, CDR2 and CDR3, where CDR1 has the amino acid sequence SEQ ID NO: 60, CDR2 has the amino acid sequence SEQ ID NO: 61 and CDR3 has the amino acid sequence SEQ ID NO: 62; or ii) hypervariable regions CDR1, CDR2 and CDR3, where CDR1 has the amino acid sequence SEQ ID NO: 63, CDR2 has the amino acid sequence SEQ ID NO: 64 and CDR3 has the amino acid sequence SEQ ID NO: 65; or iii) hypervariable regions CDR1, CDR2 and CDR3, where CDR1 has the amino acid sequence SEQ ID NO: 66, CDR2 has the amino acid sequence SEQ ID NO: 67 and CDR3 has the amino acid sequence SEQ ID NO: 68.

[0202] In one embodiment, the IL-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS comprises: a) a first immunoglobulin heavy chain variable domain (V) comprising the amino acid sequence set forth as SEQ ID NO: 85. H1 ); b) a first immunoglobulin light chain variable domain (V) comprising the amino acid sequence set forth as SEQ ID NO: 101. L1 ); c) a first immunoglobulin V comprising the amino acid sequence shown as SEQ ID NO: 85. H1 A first immunoglobulin V comprising the domain and the amino acid sequence shown as SEQ ID NO: 101. L1 domain; d) a first immunoglobulin V comprising the hypervariable regions shown as SEQ ID NO: 76, SEQ ID NO: 77 and SEQ ID NO: 78; H1domain; e) a first immunoglobulin V comprising hypervariable regions as shown in SEQ ID NO: 92, SEQ ID NO: 93 and SEQ ID NO: 94; L1 domain; f) a first immunoglobulin V comprising hypervariable regions as shown in SEQ ID NO: 79, SEQ ID NO: 80 and SEQ ID NO: 81 H1 domain; g) a first immunoglobulin V comprising hypervariable regions as shown in SEQ ID NO: 95, SEQ ID NO: 96 and SEQ ID NO: 97; L1 domain; h) a first immunoglobulin V comprising the hypervariable regions shown as SEQ ID NO: 76, SEQ ID NO: 77 and SEQ ID NO: 78; H1 A first immunoglobulin V comprising the domains and hypervariable regions shown as SEQ ID NO:92, SEQ ID NO:93 and SEQ ID NO:94. L1 Domains; i) a first immunoglobulin V comprising hypervariable regions as shown in SEQ ID NO: 79, SEQ ID NO: 80 and SEQ ID NO: 81 H1 A first immunoglobulin V comprising the domains and hypervariable regions set forth in SEQ ID NO: 95, SEQ ID NO: 96 and SEQ ID NO: 97. L1 domain; j) a first light chain comprising SEQ ID NO:103; k) a first heavy chain comprising SEQ ID NO:87; or l) a first light chain comprising SEQ ID NO:103 and a first heavy chain comprising SEQ ID NO:87.

[0203] In one embodiment, the IL-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS comprises a) a second immunoglobulin heavy chain variable domain (V) comprising the amino acid sequence set forth as SEQ ID NO: 53. H2 ); b) a second immunoglobulin light chain variable domain (V) comprising the amino acid sequence set forth as SEQ ID NO: 69. L2 ); c) a second immunoglobulin V comprising the amino acid sequence shown as SEQ ID NO: 53. H2 A second immunoglobulin V comprising the domain and the amino acid sequence shown as SEQ ID NO:69. L2 domain; d) a second immunoglobulin V comprising the hypervariable regions shown as SEQ ID NO: 44, SEQ ID NO: 45 and SEQ ID NO: 46. H2 domain; e) a second immunoglobulin V comprising the hypervariable regions shown as SEQ ID NO: 60, SEQ ID NO: 61 and SEQ ID NO: 62. L2domain; f) a second immunoglobulin V comprising the hypervariable regions shown as SEQ ID NO: 47, SEQ ID NO: 48 and SEQ ID NO: 49. H2 domain; g) a second immunoglobulin V comprising the hypervariable regions shown as SEQ ID NO:63, SEQ ID NO:64 and SEQ ID NO:65. L2 domain; h) a second immunoglobulin V comprising the hypervariable regions shown as SEQ ID NO: 44, SEQ ID NO: 45 and SEQ ID NO: 46. H2 A second immunoglobulin V comprising the domains and hypervariable regions shown as SEQ ID NO:60, SEQ ID NO:61 and SEQ ID NO:62. L2 Domains; i) a second immunoglobulin V comprising hypervariable regions as shown in SEQ ID NO: 47, SEQ ID NO: 48 and SEQ ID NO: 49 H2 A second immunoglobulin V comprising the domains and hypervariable regions shown as SEQ ID NO:63, SEQ ID NO:64 and SEQ ID NO:65. L2 domain; j) a second light chain comprising SEQ ID NO:81; k) a second heavy chain comprising SEQ ID NO:55; or l) a second light chain comprising SEQ ID NO:81 and a second heavy chain comprising SEQ ID NO:55.

[0204] In some embodiments, an IL-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS comprises the three CDRs of SEQ ID NO: 53. In other embodiments, an IL-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS comprises the three CDRs of SEQ ID NO: 69. In other embodiments, an IL-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS comprises the three CDRs of SEQ ID NO: 53 and the three CDRs of SEQ ID NO: 69. In some embodiments, an IL-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS comprises the three CDRs of SEQ ID NO: 85. In other embodiments, an IL-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS comprises the three CDRs of SEQ ID NO: 101. In other embodiments, an IL-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS comprises the three CDRs of SEQ ID NO: 85 and the three CDRs of SEQ ID NO: 101.

[0205] In some embodiments, an IL-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS comprises the three CDRs of SEQ ID NO: 85. In other embodiments, an IL-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS comprises the three CDRs of SEQ ID NO: 101. In other embodiments, an IL-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS comprises the three CDRs of SEQ ID NO: 85 and the three CDRs of SEQ ID NO: 101. In some embodiments, an IL-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS comprises the three CDRs of SEQ ID NO: 53. In other embodiments, an IL-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS comprises the three CDRs of SEQ ID NO: 69. In other embodiments, an IL-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS comprises the three CDRs of SEQ ID NO: 53 and the three CDRs of SEQ ID NO: 69. In one embodiment, an L-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS comprises the three CDRs of SEQ ID NO: 85, the three CDRs of SEQ ID NO: 101, the three CDRs of SEQ ID NO: 53 and the three CDRs of SEQ ID NO: 69.

[0206] In one embodiment, the first part of the IL-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS is selected from a human IL-18 antibody comprising at least: a) an immunoglobulin heavy chain or a fragment thereof comprising a variable domain comprising hypervariable regions CDR1, CDR2 and CDR3 in their sequence, wherein CDR1 has the amino acid sequence SEQ ID NO: 76, CDR2 has the amino acid sequence SEQ ID NO: 77 and CDR3 has the amino acid sequence SEQ ID NO: 78, and a constant part of a human heavy chain or a fragment thereof; and b) an immunoglobulin light chain or a fragment thereof comprising a variable domain comprising hypervariable regions CDR1, CDR2 and CDR3 in their sequence, wherein CDR1 has the amino acid sequence SEQ ID NO: 92, CDR2 has the amino acid sequence SEQ ID NO: 93 and CDR3 has the amino acid sequence SEQ ID NO: 94, and a constant part of a human light chain or a fragment thereof. Furthermore, the second part of the IL-18 / IL-1β bispecific antibody is selected from a human IL-1β antibody comprising at least: a) an immunoglobulin heavy chain or a fragment thereof comprising a variable domain comprising in its sequence the hypervariable regions CDR1, CDR2 and CDR3, wherein CDR1 has the amino acid sequence SEQ ID NO: 44, CDR2 has the amino acid sequence SEQ ID NO: 45 and CDR3 has the amino acid sequence SEQ ID NO: 46, and a constant part of a human heavy chain or a fragment thereof; and b) an immunoglobulin light chain or a fragment thereof comprising a variable domain comprising in its sequence the hypervariable regions CDR1, CDR2 and CDR3, wherein CDR1 has the amino acid sequence SEQ ID NO: 60, CDR2 has the amino acid sequence SEQ ID NO: 61 and CDR3 has the amino acid sequence SEQ ID NO: 62, and a constant part of a human light chain or a fragment thereof.

[0207] In one embodiment, the first part of the IL-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS is selected from a human IL-18 antibody comprising at least: a) an immunoglobulin heavy chain or a fragment thereof comprising a variable domain comprising hypervariable regions CDR1, CDR2 and CDR3 in their sequence, wherein CDR1 has the amino acid sequence SEQ ID NO: 76, CDR2 has the amino acid sequence SEQ ID NO: 77 and CDR3 has the amino acid sequence SEQ ID NO: 78, and a constant part of a human heavy chain or a fragment thereof; and b) an immunoglobulin light chain or a fragment thereof comprising a variable domain comprising hypervariable regions CDR1, CDR2 and CDR3 in their sequence, wherein CDR1 has the amino acid sequence SEQ ID NO: 92, CDR2 has the amino acid sequence SEQ ID NO: 93 and CDR3 has the amino acid sequence SEQ ID NO: 94, and a constant part of a human light chain or a fragment thereof. Furthermore, the second part of the IL-18 / IL-1β bispecific antibody is selected from a human IL-1β antibody comprising at least: a) an immunoglobulin heavy chain or a fragment thereof comprising a variable domain comprising in its sequence the hypervariable regions CDR1, CDR2 and CDR3, wherein CDR1 has the amino acid sequence SEQ ID NO: 44, CDR2 has the amino acid sequence SEQ ID NO: 45 and CDR3 has the amino acid sequence SEQ ID NO: 46, and a constant part of a human heavy chain or a fragment thereof; and b) an immunoglobulin light chain or a fragment thereof comprising a variable domain comprising in its sequence the hypervariable regions CDR1, CDR2 and CDR3, wherein CDR1 has the amino acid sequence SEQ ID NO: 60, CDR2 has the amino acid sequence SEQ ID NO: 61 and CDR3 has the amino acid sequence SEQ ID NO: 62, and a constant part of a human light chain or a fragment thereof.

[0208] The first V of the IL-18 / IL-1β bispecific antibody used in the disclosed method H1 Or V L1 The domain is represented by SEQ ID NOs: 85 and 101. H Or V L A first V that is substantially identical to the domain H1 and / or the first V L1As disclosed herein, an IL-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS may comprise a first heavy chain substantially identical to that set forth as SEQ ID NO: 87 and / or a first light chain substantially identical to that set forth as SEQ ID NO: 103. As disclosed herein, an IL-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS may comprise a first heavy chain comprising SEQ ID NO: 87 and a first light chain comprising SEQ ID NO: 103. An IL-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS may comprise: a) a first heavy chain comprising a variable domain having an amino acid sequence substantially identical to that set forth in SEQ ID NO: 85 and a constant portion of a human heavy chain having a heterodimerization modification; and b) a first light chain comprising a variable domain having an amino acid sequence substantially identical to that set forth in SEQ ID NO: 101 and a constant portion of a human light chain. The constant portion of the human heavy chain may be an IgG1. In one embodiment, the IgG1 is a human IgG1 without effector mutations. In one embodiment, the human heavy chain IgG1 comprises a silencing mutation N297A, D265A or a combination of L234A and L235A. In a specific embodiment, the human heavy chain IgG1 comprises a silencing mutation that is a combination of L234A and L235A according to SEQ ID NO:87.

[0209] The second V of the IL-18 / IL-1β bispecific antibody used in the disclosed methods H2 Or V L2 The domain is represented by SEQ ID NOs: 53 and 69. H Or V L A second V that is substantially identical to the domain H2 and / or the first V L2As disclosed herein, an IL-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS may comprise a second heavy chain substantially identical to that set forth as SEQ ID NO: 55 and / or a second light chain substantially identical to that set forth as SEQ ID NO: 71. As disclosed herein, an IL-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS may comprise a second heavy chain comprising SEQ ID NO: 53 and a second light chain comprising SEQ ID NO: 69. As disclosed herein, an IL-18 / IL-1β bispecific antibody for use in the treatment or prevention of HS may comprise a) a second heavy chain comprising a variable domain having an amino acid sequence substantially identical to that set forth in SEQ ID NO: 53 and a constant portion of a human heavy chain having a heterodimerization modification that is complementary to the heterodimerization of the first heavy chain, and b) a second light chain comprising a variable domain having an amino acid sequence substantially identical to that set forth in SEQ ID NO: 69 and a constant portion of a human light chain. The constant portion of the human heavy chain may be IgG1. In one embodiment, the IgG1 is a human IgG1 without effector mutations. In one embodiment, the human heavy chain IgG1 comprises a silencing mutation N297A, D265A or a combination of L234A and L235A. In a specific embodiment, the human heavy chain IgG1 comprises a silencing mutation that is a combination of L234A and L235A according to SEQ ID NO:55.

[0210] Other preferred IL-18 antagonists (e.g., antibodies) for use as the first part of a bispecific antibody in the disclosed methods, kits, and dosing regimens are those set forth in U.S. Pat. No. 9,376,489, which is incorporated by reference herein in its entirety.

[0211] Other preferred IL-1β antagonists (e.g., antibodies) for use as the bispecific second moiety in the disclosed methods, kits, and dosing regimens are those set forth in U.S. Pat. Nos. 7,446,175 or 7,993,878 or 8,273,350, which are incorporated by reference in their entireties.

[0212] Example 4: In vitro activity of bbmAb1 The binding activity of bbmAb1 was tested in a variety of different cellular assays.

[0213] (1) Materials and methods (a) Solution equilibrium titration (SET) assay The following materials were used: Recombinant human IL-18, biotinylated (BTP25828) Recombinant cynomolgus IL-1β (Novartis) Anti-human IgG antibody, sulfo-TAG labeled (Meso Scale discovery (MSD) #R32AJ-5) Goat anti-human Fab specific, conjugated with MSD Sulfo-TAG NHS ester (Jackson Immuno Research #109-005-097, MSD #R91AN-1) BSA (Sigma #A-9647) MSD Lead Buffer with Detergent (MSD#R92TC-1) Phosphate-buffered saline (PBS) 10x (Teknova #P0195) Tris-buffered saline, pH 7.5 (TBS) 10x (Teknova #T1680) Tween-20 (Fluka #93773) Polypropylene microtiter plates (MTP) (Greiner #781280) 384-well plate, standard (MSD#L21XA)

[0214] (b) Cell assay and SET assay mAb2 as described in the IL-1β antibody section. mAb1 as described in the IL-18 antibody section. bbmAb1 as described in Example 1. Recombinant human IL-18 (BTP25829) purchased from MBL Int. Corp. (#B001-5) Recombinant marmoset IL-1β (Novartis) Recombinant marmoset IL-18 (Novartis) Recombinant human IL-12 (#573008) was purchased from Biolegend. KG-1 cell line (ATCC#CCL-246) Normal human dermal fibroblasts (#CC-2509) were purchased from Lonza. Marmoset skin fibroblasts (#42637F(510)) HEK-Blue™ IL-18 / IL-1β cells (#hkb-il18) were purchased from InvivoGen. PBMCs were isolated from buffy coats and obtained from the Blutspendezentrum Bern. Marmoset blood was obtained from SILABE, Niederhausbergen. IL-6 ELISA: human (BioLegend, #430503); marmoset (U-CyTech biosciences, CT974-5) IFNγ ELISA: Human (BD555142) and marmoset (U-CyTech biosciences #CT340A) QUANTI-Blue™ assay (#rep-qb1) for detection of SEAP was purchased from InvivoGen. Cell culture medium: RPMI 1640 (Invitrogen #31870) supplemented with 10% fetal bovine serum (Invitrogen #10108-157), 1% L-glutamine (Invitrogen #25030-03), 1% penicillin / streptomycin (Invitrogen #15140-148), 10 μM 2-mercaptoethanol (Gibco #31350-010), 5 mM Hepes (Gibco #15630-080) Round bottom, tissue culture treated 96 well plates (Costar #3799) Flat bottom, tissue culture treated 96 well plates (Costar #3596) Ficoll-Pacque™ Plus (GE Healthcare Life Sciences #17-1440-02) PBS 1X, calcium and magnesium free (Gibco #14190094) Leucosep tubes with porous septa, 50 mL, polypropylene (Greiner bio-one #227290) Falcon 15mL polypropylene conical tube (BD#352096) Falcon 50mL polypropylene conical tube (BD#352070)

[0215] (c) Affinity measurement by SET SET individual target binding assays Twenty-two serial 1.6n dilutions of antigen (maximum concentrations: huIL-18, 5 nM; marIL-18, 10 nM; huIL-1β, 0.5 nM; marIL-1β, 0.5 nM) were prepared in sample buffer (PBS containing 0.5% bovine serum albumin (BSA) and 0.02% Tween-20) and constant concentrations of antibody were added (10 pM for IL-18 readings, 1 pM for IL-1β readings). A volume of 60 μL / well of each antigen-antibody mixture was dispensed in duplicate into a 384-well polypropylene microtiter plate (MTP). Sample buffer served as a negative control and samples containing only antibody served as positive controls (maximum electrochemiluminescence signal without antigen, B max ). The plates were sealed and incubated overnight (o / n, at least 16 hours) at room temperature (RT) on a shaker.

[0216] IL-18 readout: Streptavidin-coated 384-well MSD array MTPs were coated with 30 μL / well of biotinylated huIL-18 (0.1 μg / mL, PBS) and incubated for 1 hour at RT on a shaker.

[0217] IL-1β read: Standard 384-well MSD array MTPs were coated with 30 μL / well of huIL-1 (3 μg / mL, PBS) diluted in PBS as supplement and incubated overnight at 4°C.

[0218] Plates were blocked with 50 μL / well of blocking buffer (PBS containing 5% BSA) for 1 hour (h) at room temperature (RT). After washing (TBST, TBS containing 0.05% Tween 20), a volume of 30 μL / well of the equilibrated antigen-antibody mixture was transferred from the polypropylene MTP to the coated MSD plate and incubated for 20 min at RT. After a further washing step, 30 μL of sulfo-tag labeled anti-IgG detection antibody (0.5 μg / mL) diluted in sample buffer was added to each well and incubated for 30 min at RT on a shaker. The MSD plate was washed and 35 μL / well MSD lead buffer was added and incubated for 5 min at RT. Electrochemiluminescence (ECL) signal was developed and measured by an MSD Sector Imager 6000.

[0219] SET simultaneous target binding assay SET assays were performed as described above, except for assay A: K D Assay B: Equilibration steps (antibody / antigen mixtures) were performed in the presence of an excess of one target (either IL18 or IL-1β at 500 pM) while assessing the free antibody concentration of the IL18- and IL-1β-coated plates. Assay B: Equilibration steps (antibody / antigen mixtures) were performed with both targets in serial dilutions in one mixture at the same time (constant concentration of antibody 10 pM, maximum antigen concentration, see above). The same mixtures were then analyzed for their free antibody concentration on IL18 and IL-1β coated plates as described above.

[0220] SET data were exported to Xlfit, an MS Excel add-in software. The mean ECL-signal was calculated from duplicate measurements within each assay. Data were baseline adjusted by subtracting the lowest value from all data points and plotted against the corresponding antigen concentration to generate titration curves. K was calculated by fitting the plot with D The values ​​were determined:

[0221] 1:2 binding model for monospecific Abs

[0222]

number

number

[0223] During the ceremony, y: ECL signal subtracted from blank B max : Maximum ECL signal at zero antigen concentration [IgG]: applied antibody concentration [Fab]: total Fab concentration applied K D :dissociation equilibrium constant x: antigen concentration applied

[0224] (d) Cell culture 2x10 5 ~1x10 6 KG-1 cells were grown in RPMI 1640 supplemented with 10% fetal bovine serum, 1% L-glutamine and 1% penicillin / streptomycin at a density of 10 viable cells / mL.

[0225] Normal human fibroblasts and marmoset fibroblasts were grown in FBM (Clonetics, CC-3131) containing bFGF (1 ng / mL, CC-4065), insulin (5 μg / mL, CC-4021) and 2% FCS (CC-4101). Fibroblast basal medium (LONZA#CC-3131) was used as the starvation medium.

[0226] HEK-Blue™ IL-18 / IL-1β cells were grown in growth medium (DMEM, 4.5 g / L glucose, 10% (v / v) fetal bovine serum, 50 U / mL penicillin, 50 mg / mL streptomycin, 100 mg / mL Normocin™, 2 mM L-glutamine) supplemented with 30 μg / mL blasticidin, 200 μg / mL HygroGold™ and 100 μg / mL Zeocin™.

[0227] Human peripheral blood mononuclear cells (PBMCs) were freshly isolated from buffy coats using LeucoSep tubes according to the manufacturer's instructions. Briefly, 13 mL of Ficoll-Paque was pre-loaded into a 14 mL LeucoSep tube by centrifugation at 1,000xg for 30 seconds. Heparinized whole blood samples were diluted with an equal volume of PBS, and 25 mL of diluted blood was added to the LeucoSep tube. The cell separation tube was centrifuged at 800xg for 15 minutes at room temperature without disruption. The cell suspension layer was collected and the cells were washed twice with PBS (two consecutive washes at 640 and 470xg for 10 minutes each) and resuspended in medium before counting.

[0228] Marmoset blood was collected in heparinized tubes and filtered using a 70 μm cell strainer (BD Biosciences #352350).

[0229] (e) IL-1β neutralization assay IL-1β-induced IL-6 production assay in fibroblasts was performed essentially as described (Gram2000) with minor modifications. Briefly, fibroblasts were seeded at a density of 5x103 cells / well (in 100μL) in 96-well flat-bottom tissue culture plates. The next day, cells were starved in starvation medium for 5 hours before the addition of recombinant IL-1β / compound solution mixture (IL-1β concentrations are shown in the table). IL-1β / compound solution mixtures were prepared in advance by incubating recombinant IL-1β with a range of compound concentrations for 30 minutes at 37°C. After o / n incubation at 37°C, cell supernatants were harvested and the amount of released IL-6 was determined by ELISA. IL-1β-induced IL-6 production assay in PBMCs was performed as follows. 3x10 cells were seeded at a density of 5x103 cells / well (in 100μL). The next day, cells were starved in starvation medium for 5 hours before the addition of recombinant IL-1β / compound solution mixture (IL-1β concentrations are shown in the table). IL-1β / compound solution mixtures were prepared in advance by incubating recombinant IL-1β with a range of compound concentrations for 30 minutes at 37°C. After o / n incubation at 37°C, cell supernatants were harvested and the amount of released IL-6 was determined by ELISA. IL-1β-induced IL-6 production assay in PBMCs was performed as follows. 5PBMCs were seeded in 96-well tissue culture plates at 100 μL / well and incubated with recombinant IL-1β / compound solution mixtures for 24 hours at 37° C. (IL-1β concentrations are shown in the table). IL-1β / compound solution mixtures were prepared in advance by incubating recombinant IL-1β with a range of compound concentrations for 30 minutes at 37° C. Cell supernatants were harvested 24 hours after stimulation and the amount of released IL-6 was determined by ELISA.

[0230] (f) IL-18 neutralization assay The assay was essentially performed as follows: 3x10 5 KG-1 cells (previously starved for 1 h in PBS+1% FCS) or PBMCs were seeded in round-bottom 96-well cell culture plates at a density of 100 μL / well and incubated with a solution mixture of recombinant IL-18 / IL-12 together with a range of compound concentrations (IL-18 / IL-12 concentrations are shown in the table). After 24 h of incubation at 37° C., supernatants were harvested and the amount of released IFNγ was determined by ELISA. For assays with marmoset blood, 85 μL / well of blood was used.

[0231] (g) Dual IL1β / IL-18 neutralization in HEK-Blue™ cells The assay was performed essentially as described in the manufacturer's operating instructions. Briefly, 4x10 cells were cultured in 96-well cell culture plates. 4 HEK-Blue™ cells were seeded at a density of 1000 μg / well and incubated with a solution mixture of recombinant IL-1β and IL-18 together with a range of compound concentrations (to generate a 1:1 SEAP signal). After 24 hours of incubation at 37° C., supernatants were harvested and the amount of released SEAP was determined by using the QUANTI-Blue™ method according to the manufacturer's instructions.

[0232] All data was exported to EXCEL software and IC50 values ​​were calculated by plotting dose-response curves against a logarithmic curve fitting function using either EXCEL / XLfit4 or GraphPad Prism software.

[0233] (2) Results (a) Affinity for recombinant human and marmoset IL1β and IL-18 The binding affinity of bbmAb1 to human and marmoset recombinant IL-1β and IL-18 proteins was measured by solution equilibrium titration (SET) titration, and the resulting K D Values ​​were compared to mAb2 for IL-1β and to mAb1 for IL-18 binding.

[0234] Comparing binding affinities in individual target binding assays, bbmAb1 showed similar average KD compared to mAb1 for human and marmoset IL-18 (Table 7). For human IL-1β binding, the average KD values ​​were slightly higher for bbmAb1 (2.6 pM) compared to mAb2 (0.6 pM), but in the same low pM range. Subsequent measurements in a simultaneous dual target binding assay (Table 8) confirmed that bbmAb1 binding KD values ​​for IL-1β were similar to those of mAb2 in preclinical and clinical grade material. Thus, bbmAb1 retains binding affinity for both targets in human and marmoset similar to mAb2 and mAb1, respectively.

[0235] [Table 5]

[0236] In addition to the individual target binding results, the K D The simultaneous dual target binding affinity of bbmAb1 was investigated by applying either one target in excess during the assessment of binding (Assay A) or by applying a mixture of both targets during serial dilution (Assay B) (Table 8). The simultaneous IL-1β / IL-18 affinity determinations showed no significant differences between Assay A (one antigen in excess) and Assay B (mixture of both antigens in serial dilution), proving that both targets are bound simultaneously without affecting the binding of the other target. Furthermore, the K values ​​obtained by the simultaneous dual binding assay D Values ​​are the K values ​​obtained in standard assays.D The values ​​were similar to those in Table 7 (in the absence of a second antigen), demonstrating that bbmAb1 can bind both antigens independently. Thus, bbmAb1 binds both human IL-1β and IL-18 simultaneously and independently, and is fully cross-reactive with the corresponding marmoset proteins.

[0237] [Table 6]

[0238] (b) Neutralizing activity of bbmAb1 in human and marmoset cell assays. The neutralizing activity of bbmAb1 against both cytokines (IL1β and IL-18) was evaluated (mAb2mAb1). Furthermore, the potency of bbmAb1 for neutralizing marmoset IL-1β and IL-18 using a marmoset cell assay system was evaluated (see section d).

[0239] (c) Individual and simultaneous IL-1β and IL-18 neutralization in human cells The neutralizing activity of bbmAb1 on IL-1β was evaluated by inhibition of recombinant IL-1β-induced IL-6 production in human dermal fibroblasts (IL-1β at 6 pM) and in human PBMCs (IL-1β at 60 pM). The neutralizing activity of bbmAb1 on IL-18 was measured by inhibition of recombinant IL-18-induced IFN-γ production in KG-1 cells and human PBMCs (both cells were activated with 3 nM recombinant human IL-18 together with 1 ng / mL of recombinant human IL-12). The inhibitory potency of bbmAb1 on IL-1β and IL-18 was always compared to that of either mAb2 or mAb1, respectively. Depending on the assay, the average IC50 values ​​of bbmAb1 were in the sub-nM or single-digit nM range and were up to 2-4 times higher in direct comparisons mAb2 (for IL-1β) and mAb1 (for IL-18), respectively (Tables 9 and 10). The monovalent format of bbmAb1 compared to the bivalent format of mAb2 / mAb1, and potentially also the KiH mutation, may be the reason for this slight difference in potency of bbmAb1.

[0240] [Table 7]

[0241] [Table 8]

[0242] bbmAb1 was able to simultaneously neutralize the bioactivity of both IL-1β and IL-18 as demonstrated in HEK Blue™ reporter cells that produce SEAP in response to 1+1 stimulation with recombinant IL-1β and IL-18 (Table 11). Similar inhibition of SEAP in this assay system was only achievable with the combination of mAb2 and mAb1, but not with the use of the individual antibodies.

[0243] [Table 9]

[0244] (d) Neutralizing activity of bbmAb1 on marmoset IL-1β and marmoset IL-18 in marmoset cell assays. To determine the inhibitory activity of bbmAb1 in marmosets, a similar in vitro assay with marmoset cells was performed as in human cells but using recombinant marmoset IL-1β and IL-18 for stimulation. When evaluating inhibition of recombinant marmoset IL-1β-induced IL-6 production in marmoset skin fibroblasts, bbmAb1 demonstrated sub-nM potency with IC50 values ​​2-3 fold higher compared to mAb2 (Table 12). Testing of bbmAb1 with human skin fibroblasts stimulated with marmoset IL-1β yielded a similar inhibition profile as with human IL-6.

[0245] [Table 10]

[0246] Single- to double-digit nM IC50 values ​​for bbmAb1 confirmed its neutralizing activity against marmoset IL-18 as tested in an IFNγ production assay using marmoset blood cells (Table 13). Testing of bbmAb1 with human PBMC stimulated with marmoset IL-18 yielded a similar inhibition profile as measured by human IFNγ production.

[0247] Thus, bbmAb1 was shown to be fully cross-reactive with marmoset IL-1β and marmoset IL-18 in functional assays using marmoset responder cells.

[0248] [Table 11]

[0249] In a variety of different cellular assays, bbmAb1, a KiH-format IL-1β / IL-18 bispecific mAb, was found to retain high affinity binding and cytokine neutralization potency for the two individual targets IL-1β and IL-18 when compared to mAb, mAb2, and mAb1. The dual IL-1β and IL-18 neutralization properties of bbmAb1 were not only demonstrated for human cytokines / cells, but also for the corresponding marmoset cytokines / cells facilitating proper toxicology studies. The up to 2-4 fold higher IC50 values ​​generated in some of the cellular assays for IL-1β and IL-18 neutralization may be the result of monovalent binding of bbmAb1 as opposed to bivalent binding of mAb2 and mAb1, respectively. Nevertheless, dual cytokine neutralization by bbmAb1 may result in additive or synergistic inhibitory activity in vivo that may not be accurately represented in our in vitro cell systems.

[0250] Example 5: Stimulatory and blocking effects of IL-1β and IL-18 combinations in PBMCs Inflammasome activation-dependent cleavage of the effector cytokines IL-1β and IL-18 leads to induction of secondary pro-inflammatory mediators, promoting immune cell recruitment / activation not only systemically but also at the site of inflammation. In two different mouse models for lethal systemic inflammation (a) LPS injection model and (b) FCAS mice (activating missense mutation in NLRP3), simultaneous absence / inhibition of both IL-1β and IL-18 was more protective against lethality compared to single IL-1β or single IL-18 absence / inhibition, revealing additive or synergistic mechanisms for immune activation (Brydges 2013, van den Berghe 2014). bbmAb1 is a human / marmoset IL-1β / IL-18-reactive bispecific mAb with no cross-reactivity with rodents and therefore cannot be tested in mouse models. Therefore, to determine the additive or synergistic inhibitory effect of the IL-1β / IL-18 neutralization combination with bbmAb1, we mimicked inflammasome-dependent pathway activation in vitro upon stimulation of human PBMCs with LPS / IL-12 and performed unbiased gene expression analysis using microarrays. As a complementary activity, we also compared gene expression profiles of PBMCs from different donors stimulated with either a combination of recombinant IL-1β and recombinant IL-18 or with a single cytokine alone.

[0251] (3) Materials and methods (a) Cell culture and ELISA RPMI 1640 (Invitrogen #31870 or Gibco #61870-010) supplemented with 10% fetal bovine serum (Invitrogen #10108-157), 1% L-glutamine (Invitrogen #25030-03), 1% penicillin / streptomycin (Invitrogen #15140-148), 10 μM 2-mercaptoethanol (Gibco #31350-010), 5 mM Hepes (Gibco #15630-080). Recombinant human IL-1β was purchased from Sino Biological Inc. (#10139-HNAE-5). Recombinant human IL-18 was purchased from MBL (#B001-5). Recombinant human IL-12 was purchased from Biolegend (#573008). IFNγ ELISA: MAX Standard Set, BioLegend, #430103 or BD OptEIA Human IFNγ ELISA Set, BD#555142 IL-6 ELISA:MAX Standard Set, BioLegend, #430503 IL-26 ELISA:Cloud Clone Corp #SEB695Hu mAb2 as described in the IL-1β antibody section. mAb1 as described in the IL-18 antibody section. bbmAb1 as described in Example 1. LPS from Salmonella enterica serovar enteritidis, Sigma #L7770 PBMCs were isolated from buffy coats obtained from Blutspendezentrum Bern. Round bottom tissue culture treated 96 well plates (Costar #3799) Flat bottom tissue culture treated 96 well plates (Costar #3596) Ficoll-Pacque™ Plus (GE Healthcare Life Sciences #17-1440-02) PBS 1X, without calcium and magnesium (Gibco #14190094) Falcon 15mL polypropylene conical tube (BD#352096) Falcon 50mL polypropylene conical tube (BD#352070) LeucosepTM tube with porous septum, 50mL, Greiner bio-one #227290 Cell strainer 70μM, BD Biosciences #352350 Trypan blue, Sigma #T8154 RNA isolation, quantification and quality determination and qPCR: Nuclease-free water, Ambion #AM9938 Rnase Zap, Ambion #AM9780 1.5mL Eppendorf tube, sterile, Rnase- and Dnase-free RLT buffer, Qiagen#1015762 Rneasy Mini Kit, Qiagen#74104 RNase-Free DNase Set, Qiagen#79254 Agilent RNA 6000 Nano Kit, Agilent#5067-1511 Chip Priming Station, Agilent #5065-4401 IKA Vortex Mixer RNaseZAP®, Ambion #9780 Agilent 2100 Bioanalyzer High Capacity cDNA Reverse Transcription Kit, Applied Biosystems, #PN4374966 Nase-free, thin wall, forsted lid 0.2mL PCR tubes, Ambion #AM12225 MicroAmp Optical 384-well reaction plate, Applied Biosystems #4309849 TaqMan GenEx Master Mix, Applied Biosystems#4369514 PCR primers (Applied Biosystems)

[0252] [Table 12]

[0253] PBMC preparation: PBMCs were isolated from buffy coats by Ficoll-Paque gradient centrifugation in Leucosep tubes according to the manufacturer's instructions. Briefly, 15 mL of Histopaque was placed in a 50 mL Leucosep™ tube and centrifuged at 1300 rpm for 30 seconds at RT. Using a pipette, 30 mL of the diluted suspension of buffy coat was added on top of the Histopaque solution and centrifuged at 1000 g for 15 minutes at RT without disruption. Plasma was discarded (approximately 20 mL) and the interface ring was collected (=human PBMCs) and transferred to a 50 mL Falcon tube. The tube was filled with 50 mL of sterile PBS and centrifuged once at 1200 rpm for 5 minutes at RT. This centrifugation was repeated twice. The supernatant was gently discarded and the cells were resuspended in 50 mL of PBS with 2% FCS and 2 mM EDTA. The cell suspension was filtered using a 70 μm cell strainer and cells were counted using trypan blue staining (500 μL trypan blue + 200 μL cells + 300 μL PBS).

[0254] LPS / IL-12 stimulation of PBMC: Cytokine production in supernatants was prepared as follows: 250000 cells / well in 100 μL final volume were distributed in 96-well round-bottom plates. LPS was used at concentrations between 0.3 μg / mL and 3000 μg / mL together with recombinant IL-12 at 10 ng / mL. Supernatants were harvested after 24 hours at 37°C and 10% CO2.

[0255] RNA extraction from cell pellets was performed as follows: 3x10 cells in 1000 μL final volume 6 Cells / well were dispensed in flat-bottom 24-well plates. LPS was used at 3 μg / mL together with recombinant IL-12 at 10 ng / mL. Cells were harvested after 24 hours at 37° C. and 10% CO2.

[0256] Stimulation of PBMCs with recombinant cytokines: 7x10 cells per well of a 12-well plate in 1.5mL final complete RPMI medium. 6PBMCs were used. Recombinant cytokines were added at the following final concentrations: 10 ng / mL recombinant IL-1β, 3 nM recombinant IL-18, 1 ng / mL recombinant IL-12. Both supernatants and cells were harvested after 4 and 24 hours at 37°C and 10% CO2.

[0257] RNA isolation, quantification and quality assessment: Cells were pelleted and the pellet was dissolved in 350 μL of Qiagen RTL buffer with 2% β-mercaptoethanol and frozen at −20° C. or −80° C. until all samples for the study were collected. RNA isolation was performed using Qiagen standard protocols. Briefly, 350 μL of 70% ethanol was added in all samples before transfer to the RNeasy spin column and centrifuged at 8000 g for 15 s. After discarding the flow-through, 350 μL of Buffer RW1 was added and the column was centrifuged at 8000 g for 15 s to wash the spin column membrane. DNase I incubation mix was prepared according to the manufacturer's instructions, added to the RNeasy spin column and incubated at RT for 15 min. After washing with 350 μL and 500 μL of Buffer RW1, the RNeasy spin column was placed in a new 2 mL collection tube and centrifuged at full speed for 1 min. RNA was finally collected by adding 35 μL RNase-free water directly to the spin column membrane and centrifuging at 8000 g for 1 min to elute the RNA. The amount of RNA was measured using a Nanodrop ND-1000 and the RNA was stored at -20°C. RIN measurements were performed for RNA quality assessment according to the manufacturer's instructions. Briefly, 1 μL of RNA or ladder was pipetted onto an Agilent RNA 6000 Nano chip and measured by using an Agilent 2100 Bioanalyser.

[0258] Cytokine gene expression analysis by qPCR: The method was performed according to the manufacturer's instructions. Briefly, 400ng of RNA was reverse transcribed using the High-Capacity cDNA Reverse Transcription Kit according to the instructions. The cDNA solution was diluted 1 / 10 in RNA / DNA free water, 1 μL of cDNA was transferred to a 384-well reaction plate, and then mixed with 1 μL of 20X TaqMan® Gene Expression Assay target FAM gene and 10 μL of 2x TaqMan® Gene Expression Master Mix and 10 μL of RNA / DNA free water. The plate was placed on an Applied Biosystems ViiA™ 7 Real-Time PCR System, and the following instrument settings were used:

[0259] [Table 13]

[0260] The housekeeping genes used for this study were HPRT1 and RLP27. The following formula was used to calculate the relative expression levels of the target genes: 1)Ct[Ref]=(Ct[HPRT1]+Ct[RLP27]) / 2 2) dCt[Ref]=40-Ct[Ref] 3)dCt[Target]=Ct[Target]-Ct[Ref] 4) ddCt = dCt[Ref] - dCt[Target] 5) Relative target gene expression = 2^ddCt

[0261] Microarrays were performed as follows: Samples were processed by CiToxLAB France on Affymetrix HG_U133_Plus2 microarrays. They were RMA normalized and analyzed with GeneSpring 11.5.1 (Agilent Technologies, Santa Clara, CA). Pathway analysis was performed using Ingenuity Pathway Analysis (IPA) and Nextbio (Illumina). Two datasets were processed independently.

[0262] First, the data was subjected to standard quality control (QC) by CiToxLAB, in-house QC by using an R script (MA_AffyQC.R) in Rstudio suite and GeneSpring (PCA, hybridization controls). It was then filtered to remove unreliable expression levels: entities (probe sets) for which at least 100 percent of the samples in any one of the experimental conditions had values ​​above the 20th percentile were kept.

[0263] Differentially expressed genes (DEGs) were identified using the "filter on Volcano plot" feature in GeneSpring. Using filtered genes (20.0-100.0th percentile expression) along with an independent t-test, probe sets with corrected p-values ​​below 0.05 and fold changes above 2.0 were considered as differentially expressed. Benjamini-Hochberg multiple testing correction was used where possible, i.e., in testing with LPS (NUID-0000-0202-4150).

[0264] For cytokine stimulation experiments, synergy was calculated using the following formula: signal A+B / (signal A+signal B-control)≧1.5.

[0265] Individual signatures (or DEG lists) were used to calculate p-values ​​by Fisher's exact test, which represents the statistical significance of observing overlap between the signatures and "disease gene lists" (pathological vs. non-pathological) of public datasets. To do so, lists were uploaded to the Illumina Base Space Correlation Engine (formerly Nextbio) and compared using meta-analysis profiles for diseases and keyword searches.

[0266] All data were exported to EXCEL software and IC values ​​were calculated by plotting dose-response curves against a logistic curve fitting function using either EXCEL / XLfit4 or GraphPad Prism software. 50 Values ​​were calculated. Differences between treatment groups were analyzed by one-way ANOVA followed by Dunnett's multiple comparisons using GraphPad Prism software, and results were considered statistically significant at p<0.05.

[0267] (4) Results (a) bbmAb1 is highly effective in inhibiting LPS / IL-12-induced IFNγ production in whole blood Exposure of human whole blood to LPS supplemented with 10 ng / mL IL-12 results in an IFNγ response that is largely, but not exclusively, dependent on "native" IL-18 produced by blood cells. Addition of IL-12 enhances the LPS-induced IFNγ response, presumably by upregulating IL-18 receptors on responding cells.

[0268] In the experimental conditions used, IL-18 neutralization with mAb1 only led to an incomplete inhibition of IFNγ production, while IL-1β blockade (using mAb2) had only a minor effect on the IFNγ response. Interestingly, combined inhibition of IL-1β and IL-18 by either bbmAb1 or a combination of mAb2 and mAb1 more profoundly and completely inhibited IFNγ production compared to single cytokine neutralization. Inhibition of LPS (0.3 μg / mL) / IL-12-induced IFNγ in whole blood by bbmAb1, mAb2, mAb1 or combined mAb2 and mAb1 (Combo).

[0269] Apart from IFNγ, none of the other cytokines tested (IL-2, -4, -6, -8, -10, -13 and TNFα) were additively inhibited by the combination of IL-1β and IL-18 neutralization in our cellular assays (. The potency of bbmAb1 was in the same range as the combination of mAb2 and mAb1 (combo), taking into account the monovalent format of the bispecific molecule.

[0270] (b) IFNγ was additively inhibited by bbmAb1 (i.e., combined IL-1β / IL-18 inhibition) compared to single IL-1β or IL-18 inhibition in LPS / IL-12-activated human PBMCs To reveal further additive effects of combined IL-1β / IL-18 inhibition using bbmAb1 (apart from IFNγ), an unbiased transcriptomic evaluation was required. Since whole blood is not optimal for transcriptomic analysis, we adapted the LPS / IL-12 stimulation assay conditions described in the Materials and Methods section above to human PBMC samples. By using PBMCs from a total of nine donors, we could confirm that bbmAb1 additively inhibited IFNγ protein secretion into the supernatant of PBMCs. Relative to whole blood experiments, approximately 10-fold lower concentrations of the individual mAbs were used to inhibit IFNγ production. Importantly, a similar pattern of inhibition was revealed at the mRNA level for IFNγ, confirming the suitability of the samples for unbiased microarray-based gene expression analysis. Inhibition of LPS (0.3 μg / mL) / IL-12-induced IFNγ protein production and IFNγ gene expression by bbmAb1, mAb2 and mAb1 (each at a concentration of 10 nM) in human PBMC was demonstrated.

[0271] Affymetrix microarrays were performed on n=5 donors from PBMC sampled from the LPS / IL-12 stimulation experiment described in the Materials and Methods section above. Unfortunately, global evaluation of gene expression profiles revealed a strong LPS / IL-12 stimulation effect, and PCA showed clustering by donor, rather than by compound within stimulated or unstimulated groups. Nevertheless, comparison of LPS / IL-12 stimulated samples with stimulation+bbmAb1 for differentially expressed genes revealed a shortlist of genes downregulated by combined IL-1β / IL-18 blockade with bbmAb1 (Table 14). Apart from the strong downregulation of IFNγ genes, which revalidated our microarray data, the IL-26 gene was also a further cytokine gene additively inhibited by bbmAb1 compared to single IL-1β inhibition (by mAb2) or IL-18 inhibition (by mAb1).

[0272] [Table 14]

[0273] (c) IL-26 is another pro-inflammatory cytokine that is additively inhibited by bbmAb1 in LPS / IL-12-stimulated PBMCs. To further confirm that LPS / IL-12-driven IL-26 gene expression and protein production was most effectively inhibited by combined IL-1β / IL-18 blockade using bbmAb1, we extended the study to a total of n=9 PBMC donors and examined IL-26 gene expression by qPCR and IL-26 protein production by ELISA. ELISA largely confirmed the inhibition of IL-26 gene expression obtained using the microarray approach. Interestingly, IL-26 protein levels in the supernatant were only partially reduced at 24 hours by addition of mAb. The reason for this difference is unclear but may be related to the dynamic differences between IL-26 gene expression and protein production as well as the difference in consumption of IL-26 compared to IFNγ. Nevertheless, bbmAb1 was superior in reducing IL-26 protein levels in PBMC supernatants compared to mAb2 and mAb1.

[0274] (d) IL1β / IL18 signaling signature correlates with disease Previously established PBMC culture conditions, where recombinant IL-1β stimulation results in IL-6 production or recombinant IL-18 / IL-12 stimulation results in IFNγ production, were combined to reveal additive or synergistic downstream target genes or signatures (data not shown). PBMCs from n=4 donors sampled at two different time points (6 hours and 24 hours) were used for Affymetrix microarray evaluation for unbiased evaluation of gene expression profiles. Combinations of IL-1β and IL-18 stimulation revealed synergistically upregulated genes at 6 hours and 24 hours (data not shown). Addition of IL-12 to the IL-1β / IL-18 combination greatly enhanced the synergistic effect on the set of upregulated genes. The generated signaling signatures of single or combined IL-1β / IL-18 pathway stimulation (upregulated genes only) were used to interrogate patient-derived datasets across several autoimmune diseases. For example, correlations against public sarcoidosis data sets are identified. P-values ​​(calculated with Fisher's exact test) indicate significant correlation against several published studies comparing diseased and healthy tissues from sarcoidosis patients. Tissues include skin as well as lung, lacrimal gland and anterior orbit. Across all databases, the combination of IL1β / IL18 signaling showed the best correlation to disease, followed by IL-1β and IL-18. IL-1β / IL-18 differentially upregulated genes (DEGs) in PBMCs compared to DEGs from five different sarcoidosis tissues "disease vs. healthy."

[0275] (e) Conclusion LPS and recombinant IL-12 were used to mimic pathogen-associated molecular pattern (PAMP)-dependent NLRP3 inflammasome activation within the first 24 h of in vitro culture. It was found that the combination of IL-1β and IL-18 inhibition by using bbmAb1 acts additively to reduce / inhibit IFNγ production in LPS / IL-12-stimulated PBMCs. IL-12 was previously described to act synergistically with IL-18 to induce IFNγ production in T, B, NK cells, macrophages and dendritic cells (as reviewed by Nakanishi, 2001), but now an additional stimulatory effect of IL-1β on IFNγ production could be revealed under the experimental conditions used. Thus, co-incubation of PBMCs with LPS / IL-12 effectively drives the production of “native” IL-1β and IL-18, both of which contribute to a strong IFNγ response. By using unbiased microarray transcriptomics, additional genes were identified that were additively downregulated by combined IL-1β / IL-18 neutralization versus single IL-1β or IL-18 blockade. IL-26 was among the members of the IL-20 cytokine subfamily (IL-19, IL-20, IL-22, IL-24, and IL-26) that is conserved in most vertebrate species but absent in most rodent lineages (including mouse and rat) (Donnelly 2010). It signals through a heterodimeric receptor complex composed of IL-20R1 and IL-10R2 chains. The IL-26 receptor is predominantly expressed on non-hematological cell types, especially epithelial cells. Elevated levels of IL-26 were reported in the serum and especially in the synovial fluid of RA patients, which may act as a factor to promote Th17 cell proliferation and differentiation. Unfortunately, discovery of additional genes / pathways induced by combined blockade of IL-1β and IL-18 was hampered by the potent effects of LPS / IL-12 stimulation of PBMC samples.Nevertheless, both IFNγ and IL-26, and to some extent IL-22, were among the genes synergistically upregulated by combined stimulation with recombinant IL-1β and IL-18 in PBMCs, confirming these two factors as downstream effectors in this activation pathway. Thus, it appears that the IL-20 subfamily of cytokines (including IL-26 and IL-22) is highly dependent on simultaneous signals from IL-1β and IL-18. With due caution regarding the selectivity of individual signaling signatures and potential blocking effects, these comparisons are useful to demonstrate that individual pathways are active in diseases such as sarcoidosis.

[0276] Example 6: Inhibition of spontaneous IFNγ, TNFα and IL-2 production by bbmAb1 Punch biopsies (2 mm) were taken from surgically excised skin from nine different HS patients and cultured in 80 μL of medium (Iscove's modified Dulbecco's medium supplemented with 10% KnockOut serum replacement (Gibco) and 1% penicillin-streptomycin) in 96-well cell culture plates (flat bottom, tissue culture treated; Costar) for 24 h at 37° C. and 5% CO2, either in medium as an untreated control (FIG. 3, far left column) or in the presence of either bbmAb1 (FIG. 3, middle column) or adalimumab (FIG. 3, far right column) at a concentration of 100 μg / mL. After centrifugation of the plates, supernatants were collected from each HS biopsy and subjected to multiplex MSD (Meso Scale Discovery Platform) cytokine pro-inflammatory panel 1 (protein) arrays using an MSD plate reader according to the manufacturer's protocol. Data were normalized to individual biopsy weights and exported to GraphPad Prims software for blotting plots. Figure 3 shows that bbmAb1 reduces spontaneous IFNγ, TNFα and IL-2 production in supernatants of HS biopsies.

[0277] Example 7: Toxicity Testing The bispecific antibody bbmAb1 was well tolerated when administered sc to marmoset monkeys up to 100 mg / kg twice weekly for 26 weeks (no observed effect level (NOEL) 100 mg / kg; Cmax, ss of 3,110 μg / mL, AUC0-72h, ss of 218,000 μ·h / mL) and showed no effects on safety pharmacology (central nervous system, cardiovascular and respiratory systems), toxicology (including male reproductive system and sperm motility) or local tolerability. No effects were seen after intravenous (iv) administration at 100 mg / kg twice weekly for 26 weeks (Cmax, ss of 4570 μg / mL, AUC0-72h, ss of 261,000 μg·h / mL). Furthermore, no treatment-related effects were observed on immune cells in peripheral blood and on primary and secondary humoral immune responses upon foreign antigen challenge. In a single ascending dose (SAD) study of bbmAb1, data from the first six cohorts (A1-B1) at ascending doses of 0.1, 0.3, 1, 3, 10 mg / kg iv and 100 mg sc in a total of 48 subjects (of which 12 were placebo-treated) showed that bbmAb1 was generally well tolerated at doses up to 10 mg / kg. The Cmax and AUC of bbmAb1 increased proportionally with dose across the entire range of iv dosing (0.1 mg / kg to 10 mg / kg). The mean half-life of bbmAb1 was approximately 21-26 days. Bioavailability of the sc dose was estimated to be 70%.

[0278] Example 8: Therapeutic Uses A Randomized, Subject- and Physician-Blinded, Placebo-Controlled, and Multicenter Platform Study to Evaluate the Efficacy and Safety of bbmAb1 in Patients with Moderate-to-Severe Hidradenitis Suppurativa Details of the clinical trial design to demonstrate the efficacy of the bispecific anti-IL-1β anti-IL-18 antibody bbmAb1 are provided below.

[0279] Subject and investigator blinding allows for unbiased assessment of subjective readouts such as HS lesion counts or total HS-PGA scores, as well as adverse events.

[0280] A randomized, subject- and physician-blinded, placebo-controlled, multicenter, and parallel-group study is conducted to evaluate the efficacy, safety, and tolerability of several active treatment compounds, such as the bispecific anti-IL-1β anti-IL-18 antibody bbmAb1, in subjects with moderate to severe HS. After a screening period of approximately 5 weeks, a treatment period of 16 weeks is planned, followed by approximately 12 weeks of safety follow-up. Subjects will receive bbmAb1, 300 mg (3 injections of 1.5 mL; biweekly on days 1, 15, and 29, then monthly (Q4W) on days 57 and 85) sc or its corresponding placebo (2x1.5 mL) sc.

[0281] Subjects included in this study are adult male and female subjects aged 18-65 years (inclusive) with moderate to severe HS diagnosed with recurrent inflammatory lesions for at least 12 months. At the time of randomization (prior to dosing on Day 1), subjects must have at least 5 inflammatory lesions (abscesses and nodules) in at least 2 anatomical regions to be included. Randomization to cohorts and individual groups will be performed using a centralized Interactive Response Technology (IRT) system.

[0282] The primary clinical endpoint was the simplified HiSCR (Hidradenitis Suppurativa Clinical Response) after 16 weeks of treatment.

[0283] On Day 113 (Week 17), after safety and other assessments have been performed, all subjects will enter a follow-up period during which they will not receive any further study medication. Subjects may receive previously prohibited medications during this follow-up period if medically justified and if no potential safety concerns have been identified (after discussion with the sponsor).

[0284] Safety and efficacy assessments will be performed at follow-up visits as specified in the evaluation schedule. Pharmacokinetic (PK), pharmacodynamic (PD) and biomarker samples will also be collected. An end-of-study (EOS) visit will occur on Day 197 (Week 29) and will include a study completion evaluation followed by discharge from the study. Investigator and subject blinding will be maintained until study completion.

[0285] Approximately 40 subjects will be randomized; 30 subjects will receive the study treatment and 10 subjects will receive a matching placebo. - On Day 1, 300 mg bbmAb1 or its matching placebo (2 injections of 1.5 mL) will be administered by subcutaneous (sc) injection by trained medical staff. Clinical evaluations will be performed, as well as PK, target engagement, immunogenicity, pathway and disease biomarkers and safety assessments. Subjects will be discharged from the study site the same day after completion of all evaluations if there are no safety concerns. After the first dose, subjects should be observed at the site for immediate injection site reactions for at least 1 hour, or longer at the discretion of the investigator. - Subjects return to the study site during the loading phase of the study (Day 1 (Week 1) through Day 29 (Week 5)) to receive bbmAb1 sc every other week (Q2W; 3 doses). - bbmAb1 will then be administered at 300 mg sc every 4 weeks (Q4W; 2 doses) during the maintenance phase of the study (Day 29 (Week 5) - Day 85 (Week 13)).

[0286] During these visits, safety and select efficacy assessments will be performed and PK, target engagement, immunogenicity and pathway / disease biomarker samples will be collected.

[0287] The primary goal is to show preliminary efficacy of treatment with bispecific antibody bbmAb1 in HS subjects after 16 weeks of treatment in comparison to placebo. After the 16-week treatment period, a 12-week follow-up period will be included to observe whether the durability of effect can be maintained or increased after 16 weeks of treatment.

[0288] For this study, we selected the simplified HiSCR (modified from Kimball 2014) as the primary endpoint, defined as a 50% reduction in the total number of abscesses + inflammatory nodules without any increase in draining fistulas.

[0289] Inflammatory lesions in HS are counted as individual lesions (inflammatory nodules, abscesses, and draining fistulas) in typical anatomical areas. In addition to counting, a global assessment scale (Hidradenitis Suppurativa-Physician Global Assessment or HS-PGA) as well as a composite score (Hidradenitis Suppurativa Severity Assessment Score or SAHS) are used.

[0290] Several patient-reported outcomes will be used, including the Dermatology Life Quality Index (DLQI) and finally, a numeric rating scale (NRS) for pain will be included, as skin-related pain was the most important symptom in the subjects' perspective.

[0291] For further information on clinical evaluation: HS-PGA (Hidradenitis Suppurativa-Physician Global Assessment): This score was used exploratory purposes to assess HS and was used and described in Kimball AB, Kerdel F, Adams D, et al. (2012).

[0292] The SAHS score is a composite score (Hessam S, Scholl L, Sand M, et al (2018)) derived from collected information on inflammatory lesion count, fistula number and NRS pain. In addition, anatomical area and new or flared existing swelling are collected in both cohorts.

[0293] Cutaneous Pain-NRS (Numerical Rating Scale for Pain): The NRS for skin-related pain is used because it was used in adalimumab trials (Kimball et al. (2016)) and skin or HS-related pain is one of the greatest burdens for patients (Matusiak et al (2017)). Pain that is HS-related is recorded as the average in the last 24 hours and at its worst (in the last 24 hours).

[0294] Other patient-reported outcomes (PROs) include dimensions such as pruritus, fatigue and work disability, as well as the Dermatology Life Quality Index (DLQI) and Dermatology-Related Quality of Life (QoL) tools, with validated scores available in many countries and languages, including Patient Global Assessment.

[0295] [Table 15]

[0296] [Table 16]

[0297] Selection criteria: Male and female subjects, aged 18-65 years (inclusive), with clinically diagnosed HS at least 12 months prior to screening Patients with moderate to severe HS as assessed at screening and randomization (pre-dose on Day 1): A total of at least 5 inflammatory lesions, i.e. abscesses and / or inflammatory nodules, and Not more than 15 fistulas, and At least two anatomical regions must contain HS lesions Minimum weight at screening: 50kg (inclusive) Able to communicate well with the investigator, understand and comply with the requirements of the study, and be able and willing to complete study visits according to the study schedule.

[0298] Exclusion criteria: Use of other investigational drugs at screening or within 30 days or 5 half-lives of randomization, whichever is longer; or for a longer period if required by local regulations. WOCBP (defined as all women of physiological childbearing potential) are asked to consistently use highly effective contraception from at least 3 months before the first drug dose through 5 months after the last dose (days 225-253), if a pregnancy test is performed. Women who are pregnant or nursing (breastfeeding) at screening or randomization, where pregnancy is defined as the female's status after conception and through termination of pregnancy as confirmed by a positive hCG clinical test.

[0299] Study treatment and duration Patients assigned to the bbmAb1 arm will receive bbmAb1, 300 mg, sc or matching placebo as follows: every other week (Q2W) from day 1 (Week 1) to day 29 (Week 5) and then monthly (Q4W) until and including day 85 (Week 13).

[0300] Efficacy evaluation -Evaluation of the simplified and original Hidradenitis Suppurativa Clinical Response (HiSCR) International Hidradenitis Suppurativa Severity Score System (IHS4) Hidradenitis Suppurativa-Physician Global Assessment (HS-PGA) score and responder rate HS Inflammatory Lesion Count Hidradenitis Suppurativa Severity Rating (SAHS) For bbmAb1, the drug dosage form supplied is a "ready-to-use" buffered sterile aqueous solution. This solution contains 100 mg / mL bbmAb1 and the excipients L-histidine, sucrose and polysorbate 20, pH 6.0. The placebo control selected for this study is a solution with a matching composition of inactive excipients.

[0301] Predicted effects of bbmAb1 administration on IL-1β and free IL-18 The models used to predict the dynamics of the anti-IL-1β / IL-18 bispecific antibody and its target in serum consisted of a general competitive binding model for the IL-18 group (Yan et al., 2012) and a previously published model of canakinumab adjusted for bbmAb1 for the IL-1β group (Chakraborty et al., 2012), together with a new model describing free and total IL-18 dynamics. To predict the dynamics of IL-1β in response to application of bbmAb1, we used a model established in clinical canakinumab studies (Chakraborty et al., 2012) and updated the bbmAb1 specific PK parameters and binding affinity accordingly. To adjust IL-1β concentrations in CAPS patients, we used the synthesis and clearance parameters of this interleukin enumerated in the canakinumab clinical trials (Chakraborty et al., 2012). The model is based on in-house in vitro and published human data from free and total IL-18 serum concentrations from patients across several autoimmune diseases (Weiss et al., 2018).

[0302] On this basis, a dosing schedule of 300 mg Q2W / Q4W sc is predicted to result in a concomitant reduction in both IL-1β and IL-18 levels in serum, and effective neutralization of IL-1β as well as IL-18 is predicted (Figures 4 and 5).

[0303] Pharmacokinetics of bbmAb1 bbmAb1 is evaluated in an FiH single-dose ascending study up to 10 mg / kg iv in healthy volunteers with no drug-related SAEs. The pharmacokinetics (PK) of bbmAb1 in humans is as expected for a typical IgG1 antibody binding to a soluble ligand cytokine target, mirroring human expectations based on marmoset monkey data. bbmAb1 demonstrated a dose-linear increase in exposure (evaluated up to 10 mg / kg iv) consistent with expected human PK. Predicted human PK parameters for bbmAb1 include clearance (CL) = 0.158 L / d, volume of distribution (V d ) = 5.586 L (for a 70 kg human subject) or 0.08 L / kg; half-life (T 1 / 2 )=24.5 days. Preliminary analysis of the PK profile from the FiH study provides no evidence of accelerated clearance of bbmAb1 due to the formation of anti-drug antibodies (ADA).

[0304] Based on recent subcutaneous data from the FiH study, the bioavailability and absorption rate constants were adjusted for these findings and used in predicting subcutaneous PK.

[0305] A bbmAb1 dose of 300 mg Q2W / Q4W sc is predicted to lead to rapid and simultaneous neutralization of all systemic free IL-1β and IL-18. Exposure following 300 mg sc exceeds the in vitro IC90 for IL-1β and IL-18 for over 100 days following a single dose (Figure 6).

[0306] References Akdogan N,Dogan S,Incel-Uysal P,et al(2020)Serum amyloid A and C-reactive protein levels and erythrocyte sedimentation rate are important indicators in hidradenitis suppurativa.Arch Dermatol Res.;312(4):255-262. Andersen RK,Jemec GB(2017)Treatments for hidradenitis suppurativa.Clin.Dermatol.p.218-224. Andre R,Marescassier H,Gabay C,et al(2019)Long-term therapy with anakinra in hidradenitis suppurativa in three patients.Int J Dermatol;58(11):e208-e209. Bettoli V,Zauli S,Virgili A(2016)Oral clindamycin and rifampicin in the treatment of hidradenitis suppurativa-acne inversa:can some factors influence the response to the treatment? G Ital Dermatol Venereol p.216-8. Blok JL,Li K,Brodmerkel C,et al(2016)Ustekinumab in hidradenitis suppurativa:clinical results and a search for potential biomarkers in serum.Br.J.Dermatol.p.839-46. Byrd AS,Kerns ML,Williams DW,et al(2018)Collagen deposition in chronic hidradenitis suppurativa:potential role for CD163+ macrophages.Br.J.Dermatol.;179(3):792-794. Boergeson E,Docherty NG,Murphy M,et al(2011)Lipoxin A4 and benzo-lipoxin A4 attenuate experimental renal fibrosis.FASEB J.p.2967-79. Casseres RG,Prussick L,Zancanaro P,et al(2020)Secukinumab in the treatment of moderate to severe hidradenitis suppurativa:Results of an open-label trial.J Am Acad Dermatol.;82(6):1524-1526 Celtaxsys Inc.Press-release(2018)Celtaxsys announces results of phase 2 trial showing clinically meaningful improvement in pulmonary exacerbations in cystic fibrosis patients. Chakraborty,A,Tannenbaum,S,Rordorf,C.et al(2012)Pharmacokinetic and pharmacodynamic properties of canakinumab,a human anti-interleukin-1β monoclonal antibody.Clinical pharmacokinetics;51(6):e1-e18. Christmas P,Fox JW,Ursino SR,et al(1999)Differential localization of 5- and 15-lipoxygenases to the nuclear envelope in RAW macrophages.J.Biol.Chem.p.25594-8. Deckers IE,Kimball AB(2016)The Handicap of Hidradenitis Suppurativa.Dermatol Clin p.17-22. Dessinioti C,Zisimou C,Tzanetakou V,et al(2016)Oral clindamycin and rifampicin combination therapy for hidradenitis suppurativa:a prospective study and 1-year follow-up.Clin.Exp.Dermatol.p.852-857. Esmann S,Jemec GB(2011)Psychosocial impact of hidradenitis suppurativa:a qualitative study.Acta Derm.Venereol.p.328-32. Fimmel S,Zouboulis CC(2010)Comorbidities of hidradenitis suppurativa(acne inversa).Dermatoendocrinol;2(1):9-16.. Finlay AY,Khan GK(1994)Dermatology Life Quality Index(DLQI)--a simple practical measure for routine clinical use.Clin.Exp.Dermatol;19:210-216. Garg A,Wertenteil S,Baltz R,et al(2018)Prevalence Estimates for Hidradenitis Suppurativa among Children and Adolescents in the United States:A Gender-and Age-Adjusted Population Analysis.J.Invest.Dermatol.;138(10):2152-2156. Giamarellos-Bourboulis EJ,Argyropoulou M,Kanni T,et al(2020).Clinical efficacy of complement C5a inhibition by IFX-1 in hidradenitis suppurativa:an open-label single-arm trial in patients not eligible for adalimumab.Br J Dermatol.;183(1):176-178. Giuseppe P,Nicola P,Valentina C,et al(2018)A Case of Moderate Hidradenitis Suppurativa and Psoriasis Treated with Secukinumab.Ann Dermatol;30(4):462-464. Gottlieb A,Natsis NE,Kerdel F,et al(2020)A Phase II Open-Label Study of Bermekimab in Patients with Hidradenitis Suppurativa Shows Resolution of Inflammatory Lesions and Pain.J Invest Dermatol;140(8):1538-1545. Hessam S,Scholl L,Sand M,et al(2018)A Novel Severity Assessment Scoring System for Hidradenitis Suppurativa.JAMA Dermatol;154(3):330-335. Hessam S,Sand M,Gambichler T,et al(2015)Correlation of inflammatory serum markers with disease severity in patients with hidradenitis suppurativa(HS).J Am Acad Dermatol.;73(6):998-1005. Houriet C,Seyed Jafari SM,Thomi R,et al(2017)Canakinumab for Severe Hidradenitis Suppurativa:Preliminary Experience in 2 Cases.JAMA Dermatol.;1;153(11):1195-1197. Hunger RE,Surovy AM,Hassan AS,et al(2008)Toll-like receptor 2 is highly expressed in lesions of acne inversa and colocalizes with C-type lectin receptor.Br.J.Dermatol.;158(4):691-7. Ingram JR,Woo PN,Chua SL,et al(2016)Interventions for hidradenitis suppurativa:a Cochrane systematic review incorporating GRADE assessment of evidence quality.Br.J.Dermatol.;174(5):.970-8. International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human Use(ICH)(2016)ICH E6(R2)GCP Guideline(2016). Jaeger T,Andres C,Grosber M,et al(2013)Pyoderma gangrenosum and concomitant hidradenitis suppurativa-rapid response to canakinumab(anti-IL-1β).Eur J Dermatol.;23(3):408-410. Janse IC,Deckers IE,van der Maten AD,et al(2017)Sexual health and quality of life are impaired in hidradenitis suppurativa:a multicentre cross-sectional study.Br.J.Dermatol.;176(4):1042-1047. Jemec GB(2012)Clinical practice.Hidradenitis suppurativa.N.Engl.J.Med.;366(2):158-64. Jemec GB,Kimball AB(2015)Hidradenitis suppurativa:Epidemiology and scope of the problem.J.Am.Acad.Dermatol.;73(5 Suppl 1):S4-7. Jorgensen AR,Yao Y,Thomsen SF(2018)Therapeutic Response to Secukinumab in a 36-Year-Old Woman with Hidradenitis Suppurativa.Case Rep Dermatol Med;(Apr 16):8685136.. Kanni T,Zenker O,Habel M,et al(2018)Complement activation in hidradenitis suppurativa:a new pathway of pathogenesis? Br.J.Dermatol.;179(2):413-419 Kelly G,Sweeney CM,Tobin AM,et al(2014)Hidradenitis suppurativa:the role of immune dysregulation.Int.J.Dermatol.;53(10):1186-96. Kelly G,Hughes R,McGarry T,et al(2015)Dysregulated cytokine expression in lesional and nonlesional skin in hidradenitis suppurativa.Br J Dermatol.;173(6):1431-1439. Kimball AB,Ganguli A,Fleischer A(2018)Reliability of the hidradenitis suppurativa clinical response in the assessment of patients with hidradenitis suppurativa.J Eur Acad Dermatol Venereol.;32(12):2254-2256. Kimball AB,Jemec GB,Yang M,et al(2014)Assessing the validity,responsiveness and meaningfulness of the Hidradenitis Suppurativa Clinical Response(HiSCR)as the clinical endpoint for hidradenitis suppurativa treatment.Br.J.Dermatol.;171(6):1434-42. Kimball AB,Kerdel F,Adams D,et al(2012)Adalimumab for the treatment of moderate to severe Hidradenitis suppurativa:a parallel randomized trial.Ann.Intern.Med.;157:846-855. Kimball AB,Okun MM,Williams DA,et al(2016)Two Phase 3 Trials of Adalimumab for Hidradenitis Suppurativa.N.Engl.J.Med.;375(5):422-34. Leslie KS,Tripathi SV,Nguyen TV,et al(2014)An open-label study of anakinra for the treatment of moderate to severe hidradenitis suppurativa.J Am Acad Dermatol.;70(2):243-251. Lima AL,Karl I,Giner T,et al(2016)Keratinocytes and neutrophils are important sources of proinflammatory molecules in hidradenitis suppurativa.Br.J.Dermatol.;174(3):514-21. Ljungman P,Boeckh M,Hirsch HH,et al(2017)Disease Definitions Working Group of the Cytomegalovirus Drug Development Forum.Definitions of Cytomegalovirus Infection and Disease in Transplant Patients for Use in Clinical Trials.Clin Infect Dis.;64(1):87-91. Musilova J,Moran B,Sweeney CM,et al(2020)Enrichment of Plasma Cells in the Peripheral Blood and Skin of Patients with Hidradenitis Suppurativa.J Invest Dermatol;140(5):1091-1094 Penno CA,Jager P,Laguerre C et al(2020)Lipidomics Profiling of Hidradenitis Suppurativa Skin Lesions Reveals Lipoxygenase Pathway Dysregulation and Accumulation of Pro-Inflammatory Leukotriene B4.J Invest Dermatol.;140(12):2421-2432.. Reguiai Z,Fougerousse AC,Maccari F,et al(2020)Effectiveness of secukinumab in hidradenitis suppurativa:an open study(20 cases).J Eur Acad Dermatol Venereol.;34(11):e750-e751. Revuz J(2009)Hidradenitis suppurativa.J European Acad Dermatol Venereol;23:985-998. Riis PT,Soeby K,Saunte DM,et al(2015)Patients with hidradenitis suppurativa carry a higher systemic inflammatory load than other dermatological patients.Arch Dermatol Res.;307(10):885-9. Russo V,Alikhan A(2016).Failure of Anakinra in a Case of Severe Hidradenitis Suppurativa.J Drugs Dermatol.;15(6):772-774. Schuch A,Fischer T,Boehner A,et al(2018)Successful Treatment of Severe Recalcitrant Hidradenitis Suppurativa with the Interleukin-17A Antibody Secukinumab.Acta Derm.Venereol.;98(1):151-152. Serhan CN,Chiang N,Van Dyke TE(2008)Resolving inflammation:dual anti-inflammatory and pro-resolution lipid mediators.Nat.Rev.Immunol.;8(5):349-61. Shah A,Alhusayen R,Amini-Nik S(2017)The critical role of macrophages in the pathogenesis of hidradenitis suppurativa.Inflamm.Res.;66(11):931-945. Sharon VR,Garcia MS,Bagheri S,et al(2012)Management of recalcitrant hidradenitis suppurativa with ustekinumab.Acta Derm.Venereol.;92(3):320-1. Singer EA(2004)The necessity and the value of placebo.Sci Eng Ethics;10(1):51-6. Sun NZ,Ro T,Jolly P,et al(2017)Non-response to Interleukin-1 Antagonist Canakinumab in Two Patients with Refractory Pyoderma Gangrenosum and Hidradenitis Suppurativa.J Clin Aesthet Dermatol.;10(9):36-38. Tekin B,Salman A,Ergun T(2017)Hidradenitis suppurativa unresponsive to canakinumab treatment:A case report.Indian J Dermatol Venereol Leprol.;83(5):615-617. Thorlacius L,Theut Riis P,Jemec GBE(2018)Severe hidradenitis suppurativa responding to treatment with secukinumab:a case report.Br.J.Dermatol.;179(1):182-185. Tzanetakou V,Kanni T,Giatrakou S,et al(2016)Safety and Efficacy of Anakinra in Severe Hidradenitis Suppurativa:A Randomized Clinical Trial.JAMA Dermatol;152(1):52-59. Van der Zee HH,Laman JD,de Ruiter L,et al(2012)Adalimumab(antitumour necrosis factor-α)treatment of hidradenitis suppurativa ameliorates skin inflammation:an in situ and ex vivo study.Br.J.Dermatol;166(2):298-305. van der Zee HH,Prens EP(2013)Failure of anti-interleukin-1 therapy in severe hidradenitis suppurativa:a case report.Dermatology;226(2):97-100.Weber P,Seyed Jafari SM,Yawalkar N,et al(2017)Apremilast in the treatment of moderate to severe hidradenitis suppurativa:A case series of 9 patients.J.Am.Acad.Dermatol.;76(6):1189-1191. Weiss,ES,Girard-Guyonvarc’h,C,Holzinger,D,et al(2018)Interleukin-18 diagnostically distinguishes and pathogenically promotes human and murine macrophage activation syndrome.Blood,The Journal of the American Society of Hematology;131(13):1442-1455.8 Witte-Handel E,Wolk K,Tsaousi A,et al(2019)The IL-1 Pathway Is Hyperactive in Hidradenitis Suppurativa and Contributes to Skin Infiltration and Destruction.J Invest Dermatol.;139(6):1294-1305. Woodcock J,LaVange LM(2017)Master Protocols to Study Multiple Therapies,Multiple Diseases,or Both.N.Engl.J.Med.;377:62-70. Yan,X,Chen,Y and Krzyzanski,W(2012).Methods of solving rapid binding target-mediated drug disposition model for two drugs competing for the same receptor.Journal of pharmacokinetics and pharmacodynamics;39(5):543-560. Zarchi K,Dufour DN,Jemec GB(2013)Successful treatment of severe hidradenitis suppurativa with anakinra.JAMA Dermatol.;149(10):1192-1194. Zouboulis CC(2009)Zileuton,a new efficient and safe systemic anti-acne drug.Dermatoendocrinol;1(3):188-92. Zouboulis CC,Desai N,Emtestam L,et al(2015)European S1 guideline for the treatment of hidradenitis suppurativa / acne inversa.J Eur Acad Dermatol Venereol;29(4):619-44. Zouboulis CC,Nestoris S,Adler YD,et al(2003)A new concept for acne therapy:a pilot study with zileuton,an oral 5-lipoxygenase inhibitor.Arch Dermatol;139(5):668-70. Zouboulis CC, Tzellos T, Kyrgidis A, et al on behalf of European Hidradenitis Suppurativa Foundation Investigator Group(2017).Development and validation of the International Hidradenitis Suppurativa Severity Score System(IHS4),a novel dynamic scoring system to assess HS severity.Br.J.Dermatol.;177(5):1401-9.

[0307] Example 9: Development of a high concentration formulation (100-120 mg / mL) of bbmAb1 Eight different formulations were initially tested in a high-throughput plate-based assay. · F1 (120 mg / mL bbmAb1, 20 mM sodium succinate, pH 5.0, 220 mM sucrose, 0.04% polysorbate 20); · F2 (120 mg / mL bbmAb1, 20 mM histidine / histidine-Cl, pH 5.0, 220 mM sucrose, 0.04% polysorbate 20); · F3 (120 mg / mL bbmAb1, 20 mM histidine / histidine-Cl, pH 5.5, 220 mM sucrose, 0.04% polysorbate 20); · F4 (120 mg / mL bbmAb1, 20 mM histidine / histidine-Cl, pH 6.0, 220 mM sucrose, 0.04% polysorbate 20); · F5 (120 mg / mL bbmAb1, 20 mM histidine / histidine-Cl, pH 6.5, 220 mM sucrose, 0.04% polysorbate 20); · F6 (120 mg / mL bbmAb1, 20 mM potassium phosphate, pH 7.0, 220 mM sucrose, 0.04% polysorbate 20); · F7 (50 mg / mL bbmAb1, 20 mM histidine / histidine-Cl, pH 5.5, 220 mM sucrose, 0.04% polysorbate 20); · F8 (50 mg / mL bbmAb1, 20 mM potassium phosphate, pH 7.0, 220 mM sucrose, 0.04% polysorbate 20).

[0308] Formulations F1-F8 were tested for aggregate formation by size exclusion chromatography after 2 or 4 weeks at 40° C. The results showed a trend towards increased aggregation with increasing pH from 5 to 7, with formulations F6 and F8 showing significant aggregation at pH 7.0. See FIG. 7.

[0309] Formulations F1-F8 were also tested for degradation product formation by size exclusion chromatography. The results showed that the protein was stable in all formulations tested under the conditions tested (25° C., 4 weeks). However, LabChip analysis under non-reducing conditions showed clear fluorescence of degradation product formation at lower pH, with formulations at pH 5.0 showing significant degradation product formation. See FIG. 8.

[0310] Formulations F1-F8 were tested for acidic and basic variant formation in response to heat stress (25°C, 4W) by charge zone electrophoresis (CZE). Significant acidic variant formation was observed at high pH (F8) and significant basic variant formation was observed at low pH (F1 and F2). See Figures 9A and 9B.

[0311] The above formulations were also tested for resistance to freeze-thaw and mechanical stress (agitation), and the results further support a good stable formulation at pH 5.5, preferably 6.0, containing 50-120 mg / mL bbmAb1 (preferably 100 mg / mL bbmAb1) along with excipients (e.g., sugars such as sucrose) and surfactants (e.g., polysorbates such as polysorbate 20).

[0312] Further testing modalities included visual, turbidity (A405 nm), dynamic light scattering, microflow imaging and LysC peptide mapping by liquid chromatography mass spectrometry.

[0313] Sequence Listing Amino acid and nucleotide sequences useful for practicing the present invention are disclosed in Table 15.

[0314] [Table 17]

[0315] [Table 18]

[0316] [Table 19]

[0317] [Table 20]

[0318] [Table 21]

[0319] [Table 22]

[0320] [Table 23]

[0321] [Table 24]

[0322]

Table 25

[0323]

Table 26

[0324]

Table 27

[0325]

Table 28

[0326]

Table 29

[0327]

Table 30

[0328]

Table 31

[0329]

Table 32

[0330]

Table 33

[0331]

Table 34

[0332]

Table 35

[0333] [Table 36]

[0334] [Table 37]

[0335] [Table 38]

[0336] [Table 39]

[0337] Throughout the text of this application, in the event of any discrepancy between the text of the specification (eg, Table 15) and the Sequence Listing, the text of the specification shall control.

[0338] All patents and publications mentioned herein are hereby incorporated by reference in their entirety and for all purposes. Various embodiments of the present invention are presented below. 1. A method for the treatment or prevention of hidradenitis suppurativa (HS) in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a bispecific antibody antagonist that specifically binds to and inhibits the activity of IL-18 and IL-1β. 2. The antibody, a. a first moiety that is an immunoglobulin with a first variable light chain (VL1) and a first variable heavy chain (VH1) that specifically binds to IL1β, and a first constant heavy chain (CH1) with a heterodimerization modification; b. a second moiety that is an immunoglobulin with a second variable light chain (VL2) and a second variable heavy chain (VH2) that specifically binds IL-18, and a second constant heavy chain (CH2) with a heterodimerization modification that is complementary to the heterodimerization modification of the first constant heavy chain; 2. The method according to claim 1, comprising: 3. A method for slowing, preventing or reducing the onset of HS in a subject in need thereof comprising administering to said subject a therapeutically effective amount of a bispecific antibody antagonist that specifically binds to and inhibits the activity of IL-18 and IL-1β. 4. The antibody, a. a first moiety that is an immunoglobulin with a first variable light chain (VL1) and a first variable heavy chain (VH1) that specifically binds to IL1β, and a first constant heavy chain (CH1) with a heterodimerization modification; b. a second moiety that is an immunoglobulin with a second variable light chain (VL2) and a second variable heavy chain (VH2) that specifically binds IL-18, and a second constant heavy chain (CH2) with a heterodimerization modification that is complementary to the heterodimerization modification of the first constant heavy chain; 4. The method according to claim 3, comprising: 5. The method according to claim 2 or 4 above, wherein the first and second constant heavy chains of said bispecific antibody are human IgA, IgD, IgE, IgG or IgM, preferably IgD, IgE or IgG, such as human IgG1, IgG2, IgG3 or IgG4, preferably IgG1. 6. The first and second constant heavy chains of the bispecific antibody are IgG1, a. the first constant heavy chain has a point mutation that generates a knob structure and the second constant heavy chain has a point mutation that generates a hole structure; or b. the first constant heavy chain has a point mutation that generates a hole structure and the second constant heavy chain has a point mutation that generates a knob structure, and optionally c. the first and second constant heavy chains have a mutation that creates a disulfide bridge; 5. The method according to claim 2 or 4. 7.a. the first immunoglobulin VH1 domain of the bispecific antibody i. hypervariable regions CDR1, CDR2 and CDR3, wherein CDR1 has the amino acid sequence SEQ ID NO: 76, CDR2 has the amino acid sequence SEQ ID NO: 77 and CDR3 has the amino acid sequence SEQ ID NO: 78; or ii. hypervariable regions CDR1, CDR2 and CDR3, wherein CDR1 has the amino acid sequence SEQ ID NO: 79, CDR2 has the amino acid sequence SEQ ID NO: 80 and CDR3 has the amino acid sequence SEQ ID NO: 81; Includes; b. the first immunoglobulin VL1 domain of the bispecific antibody iii. hypervariable regions CDR1, CDR2 and CDR3, wherein CDR1 has the amino acid sequence SEQ ID NO: 92, CDR2 has the amino acid sequence SEQ ID NO: 93 and CDR3 has the amino acid sequence SEQ ID NO: 94; or iv. Hypervariable regions CDR1, CDR2 and CDR3, wherein CDR1 has the amino acid sequence SEQ ID NO: 95, CDR2 has the amino acid sequence SEQ ID NO: 96 and CDR3 has the amino acid sequence SEQ ID NO: 97; Includes; c. the second immunoglobulin VH2 domain of the bispecific antibody v. hypervariable regions CDR1, CDR2 and CDR3, wherein CDR1 has the amino acid sequence SEQ ID NO: 44, CDR2 has the amino acid sequence SEQ ID NO: 45 and CDR3 has the amino acid sequence SEQ ID NO: 46; or vi. Hypervariable regions CDR1, CDR2 and CDR3, wherein CDR1 has the amino acid sequence SEQ ID NO: 47, CDR2 has the amino acid sequence SEQ ID NO: 48 and CDR3 has the amino acid sequence SEQ ID NO: 49 Including, d. The second immunoglobulin VL2 domain of the bispecific antibody comprises: vii. hypervariable regions CDR1, CDR2 and CDR3, wherein CDR1 has the amino acid sequence SEQ ID NO: 60, CDR2 has the amino acid sequence SEQ ID NO: 61 and CDR3 has the amino acid sequence SEQ ID NO: 62; or viii. Hypervariable regions CDR1, CDR2 and CDR3, wherein CDR1 has the amino acid sequence SEQ ID NO: 63, CDR2 has the amino acid sequence SEQ ID NO: 64 and CDR3 has the amino acid sequence SEQ ID NO: 65. 7. The method according to any one of 1 to 6 above, 8.a. the first immunoglobulin VH1 domain of the bispecific antibody comprises the amino acid sequence SEQ ID NO:85; b. the first immunoglobulin VL1 domain of the bispecific antibody comprises the amino acid sequence SEQ ID NO:101; c. the second immunoglobulin VH2 domain of the bispecific antibody comprises the amino acid sequence SEQ ID NO:53; d. The method according to any one of 1 to 7 above, wherein the second immunoglobulin VL2 domain of the bispecific antibody comprises the amino acid sequence of SEQ ID NO: 69. 9.a. the first immunoglobulin heavy chain of the bispecific antibody comprises the amino acid sequence SEQ ID NO:87; b. the first immunoglobulin light chain of the bispecific antibody comprises the amino acid sequence SEQ ID NO: 103; c. the second immunoglobulin heavy chain of the bispecific antibody comprises the amino acid sequence SEQ ID NO:55; d. The method according to any one of 1 to 8 above, wherein the second immunoglobulin light chain of the bispecific antibody comprises the amino acid sequence of SEQ ID NO: 71. 10. The method according to any one of 1 to 9 above, wherein the administration route is subcutaneous or intravenous, or a combination of subcutaneous and intravenous. 11. The method according to any one of 1 to 10 above, wherein the dosage is from about 1.5 mg to about 15 mg of active ingredient per kilogram of human subject. 12. The method according to any one of 1 to 11 above, wherein the dosage is about 5 mg or 10 mg of active ingredient per kilogram of human subject. 13. The method according to any one of 1 to 12 above, wherein the dosage is about 150 mg to about 600 mg of the active ingredient, for example about 300 mg of the active ingredient. 14. The method according to any one of 1 to 13 above, wherein the antibody is administered via a loading dose and a maintenance dose. 15. The method according to any one of 1 to 14 above, wherein the loading dose is administered via a subcutaneous injection of a first dose, and the maintenance dose is administered via a subcutaneous injection of a second dose. 16. The method according to claim 15, wherein the first dose is from about 150 mg to about 600 mg of active ingredient, such as about 300 mg of active ingredient, and the second dose is from about 150 mg to about 600 mg of active ingredient, such as about 300 mg of active ingredient. 17. The method of claim 15 or 16, wherein the first dose is 150 mg, 300 mg or 600 mg of active ingredient and the second dose is 150 mg, 300 mg or 600 mg of active ingredient. 18. The method of any of claims 17 to 18, wherein the loading dose comprises at least three biweekly subcutaneous injections on days 1, 15, and 29, and the maintenance dose consists of a monthly (Q4W) subcutaneous injection starting on day 57. 19. The patient with hidradenitis suppurativa meets the following criteria: a. the patient has moderate to severe HS; b. the patient is an adult; c. the patient is an adolescent; d. the patient has an HS-PGA score of ≧3 prior to treatment with the CD40 antagonist; e. the patient has at least three inflammatory lesions prior to treatment with the CD40 antagonist; or f. The patient does not have extensive scarring as a result of HS (<10 fistulas) prior to treatment with a CD40 antagonist. 19. The method according to any one of 1 to 18 above, wherein the method is selected according to one of the following: 20. By the 16th week of treatment, the patient with hidradenitis suppurativa has: a. Simple HiSCR; b. Reduction of HS erythema; c.NRS30; d. A decrease of ≦6 as measured by DLQI; and / or e.Improvement of DLQI 20. The method according to any one of 1 to 19 above, which achieves at least one of the above. 21. Any of the methods described above in 1 to 19, wherein by week 16 of treatment, at least 40% of the patients achieve a simplified HiSCR; or at least 25% of the patients achieve an NRS30 response; or less than 15% of the patients experience HS erythema. 22. The patient receives, as early as one week after the first administration of the bispecific IL-18 and IL-1β antagonist, one of the following: a. Rapid relief of pain as measured by VAS or NRS; and b. A rapid decline in CRP as measured using a standard CRP assay 20. An antibody for use according to any one of 1 to 19 above, having at least one of the following: 23. The method according to any of 1 to 19 above, wherein the patient achieves a sustained response 3 months after the end of the treatment, as measured by inflammatory lesion count, Hidradenitis Suppurativa Clinical Response (HiSCR), Numeric Rating Scale (NRS), Modified Sartorius HS score, Hidradenitis Suppurativa-Physician Global Assessment (HS-PGA) or Dermatology Life Quality Index (DLQI). 24. A method according to any one of 1 to 19 above, wherein the patient achieves a sustained response 3 months after the end of the treatment, as measured by simple HiSCR (sHiSCR). 25. A pharmaceutical composition comprising a therapeutically effective amount of a bispecific anti-IL-18 and anti-IL-1β antibody (e.g., bbmAb1) and one or more pharma- ceutically acceptable carriers.

Claims

1. 1. A composition comprising a bispecific antibody antagonist that specifically binds to and inhibits activity of IL-18 and IL-1β for use in a method for treating or preventing hidradenitis suppurativa (HS) in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the bispecific antibody antagonist; a. the first immunoglobulin heavy chain of the bispecific antibody comprises the amino acid sequence SEQ ID NO:87; b. the first immunoglobulin light chain of the bispecific antibody comprises the amino acid sequence SEQ ID NO: 103; c. the second immunoglobulin heavy chain of the bispecific antibody comprises the amino acid sequence SEQ ID NO:55; d. the second immunoglobulin light chain of the bispecific antibody comprises the amino acid sequence SEQ ID NO:71; The route of administration is subcutaneous; the antibody is administered via a loading dose and a maintenance dose; the loading dose is administered at a loading dose and the maintenance dose is administered at a maintenance dose; wherein said loading dose is about 300 mg of active ingredient, said maintenance dose is about 300 mg of active ingredient, said loading dose comprising at least three biweekly subcutaneous injections on days 1, 15 and 29, and said maintenance dose consisting of a monthly (Q4W) subcutaneous injection starting on day 57.

2. The patient with hidradenitis suppurativa meets the following criteria: a. the patient has moderate to severe HS; b. the patient is an adult; c. the patient is an adolescent; d. the patient has an HS-PGA score of ≧3 prior to treatment with the bispecific antibody antagonist; e. the patient has at least three inflammatory lesions prior to treatment with the bispecific antibody antagonist; or f. The patient does not have extensive scarring as a result of HS (<10 fistulas) prior to treatment with the bispecific antibody antagonist. The composition of claim 1 , wherein the composition is selected according to one of the following:

3. 3. The composition of claim 1 or 2, wherein the hidradenitis suppurativa patient achieves an accelerated HiSCR by the 16th week of treatment.

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