Il-18 mimetics

Abstract

The present disclosure relates to novel, VHH-based binders of IL-18Rα and / or IL-18Rβ with favorable characteristics. Moreover, the present disclosure relates to pharmaceutical compositions comprising such a compound.

Description

[0001] IL-18 MIMETICS

[0002] FIELD OF THE INVENTION

[0003] The present disclosure relates to novel, VHH-based binders of IL-18Ra and / or IL-18RP with favorable characteristics. Moreover, the present disclosure relates to pharmaceutical compositions comprising such a compound.

[0004] BACKGROUND OF THE INVENTION

[0005] Cytokines are potent immunomodulatory proteins comprising huge therapeutic potential. Consequently, several different molecules were granted marketing approval for the treatment of different diseases (Propper and Balkwill, 2022; Pires et al., 2021; Berraondo et al., 2019).

[0006] However, the pleiotropic mechanism of action of many cytokines, their short half-life combined with often dose-limiting toxicities when administered systemically hamper their therapeutic applicability (Pires et al., 2021; Amin et al., 2014).

[0007] To address these inherent limitations, "next-generation cytokines" were generated (Zheng et al., 2022). For instance, muteins of IL-2 were engineered, which do not target the a-subunit of the IL-2 receptor (CD25) in order to eliminate the intrinsic bias of this cytokine for activating regulatory T cells (Tregs) (Klein et al., 2017; Dolgin, 2022). To optimize the poor pharmacokinetics of cytokines, fusions with the (effector silenced or attenuated) Fc portion of IgGs were engineered (Jazayeri and Carroll, 2008) as well as conjugates with polyethylene glycol (Eliason, 2001). Besides, bifunctional antibody cytokine fusion proteins were constructed aiming at accumulating the immunomodulatory function of cytokines at the site of disease (Murer and Neri, 2019; Neri and Sondel, 2016).

[0008] Another interesting approach regarding "next-generation cytokines" relies on exploiting antibody-derivatives that mimic the function of cytokines, referred to as cytokine mimetics or surrogate cytokines (Saxton et al., 2023). In this respect, bispecific antibody derivatives (diabodies) have been developed that dimerize the Erythropoietin receptor (EpoR) (Moraga et al., 2015). Intriguingly, different diabodies induced differential EpoR phosphorylation ranging from weak and partial agonism to full agonism. In addition to this, single domain antibody (sdAb)-based bispecifics were generated that activate signalling through the heterodimeric IL- 2 receptor Py (IL-2RPy). These bispecific antibodies (bsAbs) mimic the function of IL-2 but without preferential activation of Tregs via IL-2 receptor a binding (Harris et al., 2021). Garcia and co-workers recently described the engineering of VHH-derived surrogate agonists targeting IL-2RPy as well as type I IFN-mimicking bsAbs (Yen et al., 2022). Importantly, the group constructed surrogate agonists displaying functional diversification in terms of receptor downstream signalling compared to the natural cytokine. Moreover, also bsAbs were constructed that trigger agonistic activity through binding to IL-2RP and IL-10RP, a receptor heterodimer that naturally does not exist. This is clearly demonstrating that the modular assembly of bsAbs enables the generation of cytokine mimetics with tailor-made functionalities which might be versatile building blocks for drug discovery.

[0009] IL-18 is a proinflammatory cytokine belonging to the IL-1 family of cytokines that mediates signalling through heterodimerization of the receptor subunits IL-18Ra and IL-18RP (Yasuda et al., 2019; Dinarello et al., 2013). IL-18 stimulates IFN-y production in innate lymphoid cells as well as antigen-experienced T cells in synergy with IL-12 (Nakamura et al., 2020). Recombinant (r)IL-18 has been assessed in clinical trials and demonstrated a favorable toxicity profile but limited efficacy as monotherapy (Atallah-Yunes and Robertson, 2022; Robertson et al., 2006; Tarhini et al., 2009). Physiologically, the activity of IL-18 is balanced via the high- affinity neutralizing and naturally occurring IL-18 binding protein (IL-18BP) (Dinarello et al., 2013; Nakamura et al., 2020). After treatment with (r)IL-18, substantially elevated levels of IL- 18BP were found in the serum of patients (Robertson et al., 2006; Robertson et al. 2008). Ring and colleagues were able to show that IL18BP is expressed in the tumor microenvironment of several tumor types (Zhou et al., 2020). Moreover, the authors found increased IL-18BP concentrations in the serum of patients with non-small cell lung cancer which were further elevated post PD-1 or PD-L1 treatment. In the same study, the group engineered a decoyresistant IL-18 mutein that still triggered IL-18 receptor activation. This next-generation IL-18 derivative showed superior antitumor efficacy in preclinical models as monotherapy as well as in combination with immune checkpoint inhibition (Zhou et al., 2020).

[0010] IL-18 is a proinflammatory cytokine promoting NK cell activation as well as effector T cell maturation and function (Holder et al., 2022). Consequently, recombinant human (rh) IL-18 emerged as promising potential therapeutic inducing antitumor immunity, (rh) IL-18 was assessed in several clinical trials either as single agent or as combination therapy (see e.g. Tarhini et al., 2009; Robertson et al., 2008; Robertson et al., 2013; Simpkins et al., 2013; Robertson et al., 2018). (rh) IL-18 therapy has been well tolerated, however, clinical efficacy has been limited. A possible explanation for the lack of therapeutic efficacy relies in the inhibition of IL-18 by the IL-18BP decoy receptor (Dinarello et al., 2013). IL-18BP binds to an overlapping site on IL-18 with IL-18Ra albeit with much higher affinities. Accordingly, it antagonizes signalling of IL- 18 by blocking the IL-18Ra interaction with IL-18. Recently, Ring and colleagues described the generation of a decoy resistant mutein of IL-18 showing great promise for anti-cancer therapy in preclinical models (Zhou et al., 2020). This entity is currently in clinical investigations (NCT04787042).

[0011] Despite the relevance of IL- 18 in the formation of tumors and the potential in targeting IL- 18 for cancer treatment, at present approaches that allow to mimic IL-18 and its functions in a taylor-made fashion are still lacking.

[0012] Accordingly, there is a need in the art for improved IL-18 receptor agonists. Moreover, there is a need in the art for improved ways to mimic the function of IL-18 in a taylor-made fashion, e.g. specifically with respect to different potencies and magnitudes of activation of IL-18R receptor activation, different potencies and magnitudes for the release of IFN-y or resistance to inhibition by IL-18BP.

[0013] The present disclosure overcomes the above-described problems and addresses the abovedescribed needs.

[0014] SUMMARY OF THE INVENTION

[0015] The present disclosure addresses the needs described above in the section "Background of the Invention" by the different aspects and embodiments described below.

[0016] The present invention is, in part, based on the surprising observation that novel bispecific surrogate agonists can be obtained that are based on sdAbs (sdAb: single-domain antibody, also referred to herein as VHH or VHH), target both IL-18Ra and IL-18RP and mimic the functionality of IL-18. Moreover, it was found that it is possible to obtain such IL-18 mimics that at the same time are resistant to the decoy receptor IL-18BP. Moreover, it was surprisingly observed that molecules in which the overall design architecture with respect to valencies as well as to the spatial orientation of individual VHH-based paratopes is engineered within the molecule allow to obtain IL-18 mimetics in which the agonistic activities are tailor-made, e.g. with respect to their potency, the magnitude of the induced activation of IL-18R and IFN-y release or the tolerance to inhibition by IL-18BP receptor decoy.

[0017] In an aspect, the present disclosure relates to a VHH antibody domain or a fragment thereof, wherein

[0018] (a) said VHH antibody domain or fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of one VHH selected from the group consisting of VHH1, VHH2, VHH3, VHH4, VHH5, VHH6, VHH8, VHH9, VHH10 and VHH11 as shown in the Table of CDRs;

[0019] (b) said VHH antibody domain or fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 as defined in (a) with modification, wherein the modification is that the sequence of at least one of CDR1, CDR2 and CDR3 is humanized; or

[0020] (c) said VHH antibody domain or fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 as defined in (a) with modification, wherein the modification is

[0021] - the replacement, addition or deletion of up to three amino acids in CDR1,

[0022] - the replacement, addition or deletion of up to three amino acids in CDR2 and / or

[0023] - the replacement, addition or deletion of up to three amino acids in CDR3; Table of CDRs:

[0024]

[0025] In another aspect, the present disclosure relates to a VHH antibody domain or a fragment thereof, wherein

[0026] (A) said VHH antibody domain comprises the amino acid sequence of a VHH selected from the group consisting of VHH1, VHH2, VHH3, VHH4, VHH5, VHH6, VHH8, VHH9, VHHIO and VHH11 as shown in the Table of VHH Sequences;

[0027] (B) said VHH antibody domain comprises a VHH sequence as defined in (A) with modification, wherein the modification is that said sequence is humanized;

[0028] (C) said VHH antibody domain comprises a VHH sequence as defined in (A) with modification, wherein the modification is the replacement, addition or deletion of up to 25 amino acids; or

[0029] (D) said VHH antibody domain comprises a VHH sequence that is at least 75% identical to a VHH sequence referred to in (A);

[0030] Table of VHH Sequences:

[0031] In another aspect, aspect, the present disclosure relates to a VHH antibody domain or a fragment thereof, wherein

[0032] (A) said VHH antibody domain consists of the amino acid sequence of a VHH selected from the group consisting of VHH1, VHH2, VHH3, VHH4, VHH5, VHH6, VHH8, VHH9, VHH10 and VHH11 as shown in the Table of VHH Sequences;

[0033] (B) said VHH antibody domain consists of a VHH sequence as defined in (A) with modification, wherein the modification is that said sequence is humanized;

[0034] (C) said VHH antibody domain consists of a VHH sequence as defined in (A) with modification, wherein the modification is the replacement, addition or deletion of up to 25 amino acids; or

[0035] (D) said VHH antibody domain consists of a VHH sequence that is at least 75% identical to a VHH sequence referred to in (A). In another aspect, aspect, the present disclosure relates to a VHH antibody domain or a fragment thereof, wherein

[0036] (d) said VHH antibody domain or fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of one VHH selected from the group consisting of VHH12, VHH13, VHH14, VHH15, VHH16, VHH17, VHH18, VHH20, VHH21 and VHH22 as shown in the Table of CDRs;

[0037] (e) said VHH antibody domain or fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 as defined in (d) with modification, wherein the modification is that the sequence of at least one of CDR1, CDR2 and CDR3 is humanized; or

[0038] (f) said VHH antibody domain or fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 as defined in (d) with modification, wherein the modification is

[0039] - the replacement, addition or deletion of up to three amino acids in CDR1,

[0040] - the replacement, addition or deletion of up to three amino acids in CDR2 and / or

[0041] - the replacement, addition or deletion of up to three amino acids in CDR3.

[0042] In another aspect, aspect, the present disclosure relates to a VHH antibody domain or a fragment thereof, wherein

[0043] (E) said VHH antibody domain comprises the amino acid sequence of a VHH selected from the group consisting of VHH12, VHH13, VHH14, VHH15, VHH16, VHH17, VHH18, VHH20, VHH21 and VHH22 as shown in the Table of VHH Sequences;

[0044] (F) said VHH antibody domain comprises a VHH sequence as defined in (E) with modification, wherein the modification is that said sequence is humanized;

[0045] (G) said VHH antibody domain comprises a VHH sequence as defined in (E) with modification, wherein the modification is the replacement, addition or deletion of up to 25 amino acids; or

[0046] (H) said VHH antibody domain comprises a VHH sequence that is at least 75% identical to a VHH sequence referred to in (E).

[0047] In another aspect, aspect, the present disclosure relates to a VHH antibody domain or a fragment thereof, wherein (E) said VHH antibody domain consists of the amino acid sequence of a VHH selected from the group consisting of VHH12, VHH13, VHH14, VHH15, VHH16, VHH17, VHH18, VHH20, VHH21 and VHH22 as shown in the Table of VHH Sequences;

[0048] (F) said VHH antibody domain consists of a VHH sequence as defined in (E) with modification, wherein the modification is that said sequence is humanized;

[0049] (G) said VHH antibody domain consists of a VHH sequence as defined in (E) with modification, wherein the modification is the replacement, addition or deletion of up to 25 amino acids; or

[0050] (H) said VHH antibody domain consists of a VHH sequence that is at least 75% identical to a VHH sequence referred to in (E).

[0051] In another aspect, aspect, the present disclosure relates to a compound comprising

[0052] - a VHH antibody domain or fragment thereof according to any one of embodiments 1 to

[0053] 188.

[0054] In another aspect, aspect, the present disclosure relates to a compound comprising

[0055] - a VHH antibody domain or fragment thereof according to any one of claims 189 to 374.

[0056] In another aspect, aspect, the present disclosure relates to a compound comprising

[0057] - a first binding module which is a VHH antibody domain or fragment thereof, wherein

[0058] (a) said VHH antibody domain or fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of one VHH selected from the group consisting of VHH1, VHH2, VHH3, VHH4, VHH5, VHH6, VHH8, VHH9, VHH10 and VHH11 as shown in the Table of CDR;

[0059] (b) said VHH antibody domain or fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 as defined in (a) with modification, wherein the modification is that the sequence of at least one of CDR1, CDR2 and CDR3 is humanized; or

[0060] (c) said VHH antibody domain or fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 as defined in (a) with modification, wherein the modification is

[0061] - the replacement, addition or deletion of up to three amino acids in CDR1, - the replacement, addition or deletion of up to three amino acids in CDR2 and / or

[0062] - the replacement, addition or deletion of up to three amino acids in CDR3;

[0063] - a second binding module which is a VHH antibody domain or a fragment thereof, wherein

[0064] (d) said VHH antibody domain or fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of one VHH selected from the group consisting of VHH12, VHH13, VHH14, VHH15, VHH16, VHH17, VHH18, VHH20, VHH21 and VHH22 as shown in the Table of CDRs;

[0065] (e) said VHH antibody domain or fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 as defined in (d) with modification, wherein the modification is that the sequence of at least one of CDR1, CDR2 and CDR3 is humanized; or

[0066] (f) said VHH antibody domain or fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 as defined in (d) with modification, wherein the modification is

[0067] - the replacement, addition or deletion of up to three amino acids in CDR1,

[0068] - the replacement, addition or deletion of up to three amino acids in CDR2 and / or

[0069] - the replacement, addition or deletion of up to three amino acids in CDR3.

[0070] In another aspect, aspect, the present disclosure relates to a compound comprising

[0071] - a first binding module which is a VHH antibody domain or fragment thereof, wherein

[0072] (A) said VHH antibody domain comprises the amino acid sequence of a VHH selected from the group consisting of VHH1, VHH2, VHH3, VHH4, VHH5, VHH6, VHH8, VHH9, VHH10 and VHH11 as shown in the Table of VHH Sequences;

[0073] (B) said VHH antibody domain comprises a VHH sequence as defined in (A) with modification, wherein the modification is that said sequence is humanized; 258 (C) said VHH antibody domain comprises a VHH sequence as defined in (A) with

[0074] 259 modification, wherein the modification is the replacement, addition or

[0075] 260 deletion of up to 25 amino acids; or

[0076] 261 (D) said VHH antibody domain comprises a VHH sequence that is at least 75%

[0077] 262 identical to a VHH sequence referred to in (A);

[0078] 263 - a second binding module which is a VHH antibody domain or a fragment thereof, wherein

[0079] 264 (E) said VHH antibody domain comprises the amino acid sequence of a VHH

[0080] 265 selected from the group consisting of VHH12, VHH13, VHH14, VHH15,

[0081] 266 VHH16, VHH17, VHH18, VHH20, VHH21 and VHH22 as shown in the

[0082] 267 Table of VHH Sequences;

[0083] 268 (F) said VHH antibody domain comprises a VHH sequence as defined in (E) with

[0084] 269 modification, wherein the modification is that said sequence is humanized;

[0085] 270 (G) said VHH antibody domain comprises a VHH sequence as defined in (E) with

[0086] 271 modification, wherein the modification is the replacement, addition or

[0087] Til deletion of up to 25 amino acids; or

[0088] 273 (H) said VHH antibody domain comprises a VHH sequence that is at least 75%

[0089] 274 identical to a VHH sequence referred to in (E).

[0090] U

[0091] 276 In another aspect, aspect, the present disclosure relates to a compound comprising

[0092] 277 - a first binding module which is a VHH antibody domain or fragment thereof, wherein

[0093] 278 (A) said VHH antibody domain consists of the amino acid sequence of a VHH

[0094] 279 selected from the group consisting of VHH1, VHH2, VHH3, VHH4, VHH5,

[0095] 280 VHH6, VHH8, VHH9, VHH10 and VHH11 as shown in the Table of VHH

[0096] 281 Sequences;

[0097] 282 (B) said VHH antibody domain consists of a VHH sequence as defined in (A)

[0098] 283 with modification, wherein the modification is that said sequence is

[0099] 284 humanized;

[0100] 285 (C) said VHH antibody domain consists of a VHH sequence as defined in (A)

[0101] 286 with modification, wherein the modification is the replacement, addition or

[0102] 287 deletion of up to 25 amino acids; or

[0103] 288 (D) said VHH antibody domain consists of a VHH sequence that is at least 75%

[0104] 289 identical to a VHH sequence referred to in (A);

[0105] 290 - a second binding module which is a VHH antibody domain or a fragment thereof, wherein (E) said VHH antibody domain consists of the amino acid sequence of a VHH selected from the group consisting of VHH12, VHH13, VHH14, VHH15, VHH16, VHH17, VHH18, VHH20, VHH21 and VHH22 as shown in the Table of VHH Sequences;

[0106] (F) said VHH antibody domain consists of a VHH sequence as defined in (E) with modification, wherein the modification is that said sequence is humanized;

[0107] (G) said VHH antibody domain consists of a VHH sequence as defined in (E) with modification, wherein the modification is the replacement, addition or deletion of up to 25 amino acids; or

[0108] (H) said VHH antibody domain consists of a VHH sequence that is at least 75% identical to a VHH sequence referred to in (E).

[0109] In another aspect, the present disclosure relates to a compound which is a bispecific antibody molecule comprising an IL-18Ra-binding VHH with an amino acid sequence as defined in the present disclosure below (First binding module) and an IL18RP-binding VHH with an amino acid sequence as defined in the present disclosure below (Second binding module).

[0110] In another aspect, the present disclosure relates to a bispecific antibody molecule prepared by the SEED (strand-exchange engineered domain) technology, said molecule comprising

[0111] - an AG chain linked to an IL-18Ra-binding VHH, wherein said AG chain linked to said IL-18Ra-binding VHH has an amino acid sequence as defined in the present disclosure below ("VHH-SEED-AG-1") and

[0112] - a GA chain linked to an IL18RP-binding VHH, wherein said GA chain linked to said

[0113] IL18RP-binding VHH has an amino acid sequence as defined in the present disclosure below ("VHH-SEED-GA-1").

[0114] In another aspect, the present disclosure relates to a pharmaceutical composition comprising the compound or bispecific antibody molecule according to the present disclosure.

[0115] In another aspect, the present disclosure relates to the use of a compound or bispecific antibody molecule according to the present disclosure for activating IL-18R receptor. In another aspect, the present disclosure relates to the use of a compound or bispecific antibody molecule according to the present disclosure for inducing IL-y release from cells carrying an IL-18R receptor.

[0116] BRIEF DESCRIPTION OF THE FIGURES

[0117] In the following, reference is made to the figures. All methods referred to in the figure descriptions below were carried out as described in detail in the examples.

[0118] Figure 1 summarizes the overall strategy applied in the present disclosure for the generation of tailor-made cytokine mimetics based on sdAb-derived bispecifics and YSD-enabled antibody discovery. (A) Bottom / Left panel: IL-18 (blue, §) triggers IL-18R downstream signalling by consecutive binding to IL-18Ra (yellow, *; the two "lobes" of IL-18Ra are each labelled with *) followed by IL-18RP (lime, #) recruitment. IL-18BP (dark grey, $) inhibits IL-18 by high- affinity binding and hence, by blocking the IL-18 / IL-18Ra interaction. Top / Right panel: Upon reformatting camelid-derived sdAbs with IL-18Ra-targeting functionality (orange, &) and IL- 18RP-targeting functionality (green, J) into an IgG-like bispecific, resulting cytokine mimetics cross-link the IL18R subunits IL-18Ra (yellow, *; the two "lobes" of IL-18Ra are each labelled with *) and IL-18RP (lime, #) and hence elicit down-stream signalling. Essentially, generated IL-18 mimetics are resistant to inhibition by IL-18BP (dark grey, $). Generated with PyMOL software version 2.3.0 based on PDB entries YYY and ZZZ and modified using www.biorender.com (B) Immunization of cam elids followed by YSD facilitate the enrichment of sdAbs specific to (rh) IL-18Ra and (rh) IL-18RP. One sublibrary was generated for each specimen (huarizo and llama) and sorted separately. In the first round of selection, a mixture of both (rh) receptor subunits was exploited at a concentration of 250 nM. In the subsequent second sorting round, enriched libraries were sorted separately against each antigen at 250 nM. A two-dimensional sorting strategy was applied to select for full-length VHH display in addition to antigen binding. Percentage of cells in sorting gates are shown. Plots show 5xl04events. (C) Graphical alignments of 55 unique sdAb clones addressing IL-18Ra (top) as well as 101 independent clones targeting IL-18RP (bottom) retrieved from YSD library sorting. Complementarity-determining regions (CDRs) are highlighted. Red bars indicate high sequence diversity at the amino acid level and green bars represent high sequence conservation at a given position. Alignment conducted with MUSCLE alignment tool using Geneious Prime 2021.1.1.

[0119] Figure 2 shows data obtained with a reporter assay, confirming that combinatorial reformatting of monospecific (1+0) SEEDbodies into strictly monovalent (1+1) bsAbs enables the identification of IL- 18 mimetics with attenuated capacities to trigger NFKB reporter activity on IL-18 reporter cells. (A) HEK-Blue™ reporter cells were incubated with increasing conentrations of reformatted bsAbs, as exemplarily shown for IL18R_VHHa2pi5, IL18R_VHHa8pi5, IL18R_VHHa2pi7 and IL18R_VHHa8pi7. Secreted embryonic alkaline phosphatase activity was monitored by determining the OD640. Reporter activity was normalized to maximal IL-18 read-out. (B) Heatmap of NFKB reporter activitation elicited by combinatorial reformatted (1+1) bsAbs. Molecules failing initial quality control (target monomer peak in SEC < 96% post protein A purification given in dark grey, functionally inactive bsAbs shown in red (J), minimally active surrogate agonists (NFKB reporter activitation < 50% compared to (rh) IL-18at 1 nM or EC50 > 0.1 nM) in yellow (#) and moderately active IL-18 mimetics (NFKB reporter activitation > 50% compared to (rh) IL-18at 1 nM or EC50 < 0.1 nM) depicted in green (*).

[0120] Figure 3 depicts data from an IFN-y release assay, confirming that bispecific (1+1) surrogate agonists trigger IFN-y release on PBMCs isolated from healthy donors. (A) IFN-y production of PBMCs stimulated with bsAbs at a fixed concentration of 100 nM or with (rh) IL-18 at 1 nM. Experiments were performed in the presence of 10 ng / ml (rh) IL-12. Graph shows box and whisker plots as superimpositions with dot plots of IFN-y release of ten different donors. ****p < 0.0001,***p < 0.001, **p < 0.01, *p < 0.05. IL18R_VHHaipi6 was used as negative control (given in red). Four leading candidates used for further characterization shown in green, purple, blue and orange. (B) TOP4 candidates evoke a dose dependent IFN-y read-out on PBMCs in the presence of low dose (rh) IL-12 (10 ng / ml). IL18R_VHHaipi6 as negative control shown in red was used at a fixed concentration of 1 pM. Mean values ± SEM of ten independent experiments are shown.

[0121] Figure 4 shows schematic depictions of the architecture of antibodies prepared and summarizes data to characterize such antibodies. These data confirm that antibody engineering enables the generation of IL-18 mimetics with augmented agonism capacities. (A) Schematic depiction of main different bispecific antibody architectures that were constructed within this work. Fusion of an anti-IL18Ra VHHa2 to the hinge region of the AG chain of the SEEDbody as well as engraftment of an anti-IL18Rp VHHP15 onto the GA chain results in the initially generated (1+1) format IL18R_VHHa2pi5. Replacing the VH and VLz. of an effector silenced IgG by VHHa2 and VHHpi5, respectively facilitates the generation of the IL18R_sdIgGa2pi5 architecture (2+2). Within the IL18R_tanVHHa2pi5 design (2+2), VHHa2 and VHHP15 are arranged in tandem (from A-terminus to C-terminus) and separated by a five amino acid Gly4Ser linker. The tandem is fused to the hinge region of an effector silenced IgGl Fc fragment. Of note, also the opposite orientation was constructed (VHHpi5 followed by VHHa2, IL18R_tanVHHpi5a2). In addition, all four molecules were also produced harboring the E430G mutation for on-target hexamerization. Structure models generated with PyMOL software version 2.3.0. (B) Distinct surrogate agonist formats of the same paratopes (VHHa2 and VHHP15) display differential properties in eliciting a functional IFN-y response on human PBMCs isolated from healthy donors at fixed concentrations. Experiments were performed at two different concentrations (10 nM and 1 nM) in the presence of 10 ng / ml (rh) IL-12. Graph shows box and whisker plots as superimpositions with dot plots of IFN-y release of six different donors. ****p < 0.0001,***p < 0.001, **p < 0.01, *p < 0.05. (C) Surrogate agonists arranged in tandem (IL18R_tanVHHa2pi5 and IL18R_tanVHHpi5a2) elicit enhanced IFN-y production in terms of potencies and magnitude on PBMCs isolated from healthy donors, resulting in a variant with increased potencies compared with (rh) IL-18. All experiments were performed in the presence of low dose (rh) IL-12 (10 ng / ml). IL18R_VHHaipi6 as negative control shown in red was used at a fixed concentration of 1 pM. Mean values ± SEM of 13 independent experiments are shown. (D) Potency augmented tandem IL- 18 mimetics are resistant to inhibition by (rh) IL-18BP, whereas (rh) IL-18 is efficiently blocked from signalling. PBMCs of healthy human donors were stimulated either with (rh) IL- 18 or IL18R_tanVHHa2pi5 and IL18R_tanVHHpi5a2 at a fixed concentration of 0.5 nM in the presence of (rh) IL-12 (10 ng / ml) and different concentrations of (rh) IL-18BP. Five independent experiments were performed and mean values ± SEM are shown.

[0122] Figure 5 summarizes the sorting procedure applied to obtain sdABs. YSD enables the enrichment of sdAbs targeting (rh) IL-18Ra and (rh) IL-18RP. Sorting output after two sorting rounds of each library against (rh) IL-18Ra and (rh) IL-18RP. A two-dimensional sorting strategy was applied do detect full-length VHH display simultaneous to the binding functionality at a concentration of 250 nM. Plots show 5xl03events of the corresponding sorting output to visualize enrichment.

[0123] Figure 6 depicts the results of binding expreiments examining if monospecific (1+0) VHH SEEDbodies show specific binding to (rh) IL-18Ra (red) or IL-18RP (green). (A) Schematic depiction of monospecific VHHs targeting (rh) IL-18Ra (red) or (rh) IL-18RP (green) engrafted onto the hinge region of the SEED backbone. sdAbs targeting IL-18Ra were fused to the hinge region of the AG chain (msVHHaX), while VHHs addressing the IL-18RP subunit were grafted onto the GA chain (msVHHpX). (B) Binding of respective monospecific VHH SEEDbodies to (rh) IL-18Ra (red) or (rh) IL-18RP (green) as determined by BLI. Either (rh) IL-18Ra or (rh) IL-18RP were loaded to HIS IK biosensors at a concentraion of 3 pg / ml followed by a first association of monospecific VHH SEEDbodies at 100 nM for 300 s. Subsequently, dissociation was measured for 100 s in kinetics buffer. Schemes generated via www.biorender.com.

[0124] Figure 7 presents data from a reporter assay suggesting that combinatorial reformatting of monospecific (1+0) SEEDbodies into strictly monovalent (1+1) bsAbs enables the identification of IL- 18 mimetics with attenuated capacities to trigger NFKB reporter activity on IL-18 reporter cells. HEK-Blue™ reporter cells were incubated with increasing conentrations of reformatted bsAbs or (rh) IL-18 and secreted embryonic alkaline phosphatase activity was monitored by determining the OD640. Reporter activity was normalized to maximal IL-18 readout. (A) Exemplary dose-dependent IL-18R agonism triggered by surrogate agonists IL18R_VHHa2pi5, IL18R_VHHa8pi5, IL18R_VHHa2pi7 and IL18R_VHHa8pi7 in direct comparison to (rh) IL-18. (B) Dose-dependent receptor agonism mediated by all 44 cytokine mimetics.

[0125] Figure 8 depicts data from an IFN-y release assay, showing that bispecific (1+1) surrogate agonists do not elicit IFN-y release on PBMCs isolated from healthy donors without the presence of low dose (rh) IL-12. IFN-y production of PBMCs stimulated with bsAbs at a fixed concentration of 100 nM or with (rh) IL-18 at 1 nM in the absence of (rh)IL-12. Graph dot plots of IFN-y release of eight different donors. IL18R_VHHaipi6 was used as negative control (given in red). Four leading candidates used for further characterization shown in green, purple, blue and orange. Figure 9 shows binding kinetics data obtained with bispecific (1+1) TOP4 surrogate agonists against receptor subunits IL-18Ra and IL-18RP. IL-18 mimetics were loaded on AHC biosensor tips at a concentration of 5 pg / ml. Interactions against both receptor subunits were measured at a concentration of 100 nM, 50 nM, 25 nM and 12.5 nM for 180 s followed by a dissociation in kinetics buffer for 300 s.

[0126] Figure 10 summarizes analytical size exclusion chromatography profiles of the generated surrogate agonist formats, indicating adequate purities following downstream purification. SEC-HPLC profiles of different formats as well as the corresponding target monomer peaks are shown. Absorbance at 214 nm was used for purity determination. (A) SEC profiles of all eight formats post protein A purification, indicating purities below 90% target species for IL18R_VHHa2pi5_E430G and IL18R_sdIgGa2pi5_E430G. (B) Analysis of IL18R_VHHa2pi5_E430G and IL18R_sdIgGa2pi5_E430G following second step SEC purification.

[0127] Figure 11 depicts data obtained by differential scanning fluorimetry to characterize the thermal unfolding of IL-18R agonistic bsAbs in different formats. Overlays of the melting curves for different formats of IL-18R agonists were recorded utilizing a temperature gradient from 20°C to 95°C at a slope of l°C / min. First derivatives of 350 nm / 330 nm curves are shown.

[0128] Figure 12 shows data obtained upon stimulation of PBMCs with the full dose-range of compound concentrations, indicating a strong hooking effect for (rh) IL 18 while for the different surrogate agonist formats it is not as pronounced. IFN-y read-out on PBMCs in the presence of low dose (rh) IL-12 (10 ng / ml). IL18R_VHHaipi6 as negative control shown in red was used at a fixed concentration of 1 pM. Mean values ± SEM of 13 independent experiments are shown.

[0129] Figure 13 shows data examining the binding of engineered tandem IL-18 mimetics. Such engineered tandem IL-18 mimetics exhibit strong binding to both receptor subunits in an avidity driven setting, (rh) IL-18Ra or (rh) IL-18RP were loaded onto HIS1K sensor tips at 5 pg / ml. A first association step was performed with different surrogate agonist formats at 100 nM, 33.3 nM and 11.1 nM for 180 s followed by a dissociation step in kinetics buffer fpr 300 s. Figure 14 shows biolayer interferometry (BLI) binding data for the binding of bispecific surrogate agonists to (rh)IL-18BP. Bispecific surrogate agonists do not bind to (rh)IL-18BP as determined, whereas (rh) IL-18BP binds to (rh)IL-18 with high affinities. BsAbs were loaded on AHC biosensor tips at 5 pg / ml. Subsequently, association was monitored using 1 pM IL- 18BP for 180 s, followed by dissociation in kinetics buffer for 180 s.

[0130] Figure 15 summarizes binding data showing that bispecific cytokine mimetics compete with (rh)IL-18 for binding to (rh)IL-18Ra. Surrogate agonists were loaded on AHC biosensor tips, followed by association using IL-18Ra at 200 nM for 300 s. Subsequently, a second association step was performed for 180 s using either (rh)IL-18 at 100 nM (red) or kinetics buffer (black).

[0131] SUMMARY OF SEQUENCES (in case of any difference between the below table and the cofiled WIPO ST.26 compliant sequence listing, the information of the below table shall prevail)

[0132]

[0133] SEQ ID NO: 1 to 11 provide the amino acid sequences of VHH1 to VHH11 (corresponding to al to al 1 according to the present disclosure). The CDRs of these VHHs are provided in SEQ ID NO: 23 to 55.

[0134] SEQ ID NO: 12 to 22 provide the amino acid sequences of VHH12 to VHH22 (corresponding to pi2 to P22 according to the present disclosure). The CDRs of these VHHs are provided in SEQ ID NO: 56 to 88.

[0135] SEQ ID NO: 89 to 99 provide the amino acid sequences of SEED AG chains based on VHH1 to VHH11 (al to al 1) according to the present disclosure. SEQ ID NO: 100 to 110 provide the amino acid sequences of SEED GA chains based on VHH12 to VHH22 (P 12 to P22) according to the present disclosure.

[0136] DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS

[0137] Although the present disclosure is described in detail above and below, it is to be understood that this disclosure is not limited to the particular methodologies, protocols and reagents described by the present disclosure, as these may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present disclosure which will be limited only by the appended claims. Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art.

[0138] In the following, certain elements of the present disclosure will be described in more detail, including the description of specific embodiments. However, the variously described examples and preferred embodiments should not be construed to limit the present disclosure to only the explicitly described embodiments. This description should be understood to support and encompass embodiments which combine the explicitly described embodiments with any number of the disclosed and / or preferred elements and in any manner. Furthermore, any permutations and combinations of all described elements in this application should be considered disclosed by the description of the present application except for where this leads to logical contradictions or the context indicates otherwise.

[0139] Unless defined otherwise herein, scientific and technical terms used in connection with the present disclosure shall have the meanings that are commonly understood by those of ordinary skill in the art. Generally, nomenclatures and techniques referred to in the present disclosure, e.g. nomenclatures and techniques of organic chemistry, chemical synthesis, biology, medicinal and pharmaceutical chemistry, medicine, pharmacology or toxicology, are those well-known and commonly used in the art. The methods and techniques of the present disclosure are generally performed according to conventional methods well-known in the art and as described in the references cited and discussed throughout the present disclosure unless otherwise indicated.

[0140] First aspect of the present disclosure (also referred to as "Embodiment 1"): According to a first aspect, the present disclosure relates to a VHH antibody domain or a fragment thereof, wherein

[0141] (a) said VHH antibody domain or fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of one VHH selected from the group consisting of VHH1, VHH2, VHH3, VHH4, VHH5, VHH6, VHH8, VHH9, VHH10 and VHH11 as shown in the Table of CDRs;

[0142] (b) said VHH antibody domain or fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 as defined in (a) with modification, wherein the modification is that the sequence of at least one of CDR1, CDR2 and CDR3 is humanized; or

[0143] (c) said VHH antibody domain or fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 as defined in (a) with modification, wherein the modification is

[0144] - the replacement, addition or deletion of up to three amino acids in CDR1,

[0145] - the replacement, addition or deletion of up to three amino acids in CDR2 and / or

[0146] - the replacement, addition or deletion of up to three amino acids in CDR3;

[0147] Table of CDRs:

[0148]

[0149] An "antibody" is a polypeptide substantially encoded by an immunoglobulin gene or immunoglobulin genes, or antigen binding fragment thereof, which specifically binds and recognizes an analyte (antigen). Immunoglobulin genes include the kappa, lambda, alpha, gamma, delta, epsilon and mu constant region genes, as well as the myriad immunoglobulin variable region genes.

[0150] In primates such as humans, a heavy and the light chain variable domain of an antibody combine to specifically bind the antigen. Generally, a naturally occurring primate (e.g., human) or murine immunoglobulin has heavy (H) chains and light (L) chains interconnected by disulfide bonds. There are two types of light chain, lambda ( ) and kappa (K). There are five main heavy chain classes (or isotypes) which determine the functional activity of an antibody molecule: IgM, IgD, IgG, IgA and IgE. Primate antibodies can be class switched.

[0151] Certain IgG antibodies from members of the camel and dromedary (Camelus bactrianus and Calelus dromaderius) family including new world members such as llama species (Lama paccos. Lama glama and Lama vicugna) of mammals as found in nature lack light chains, and are thus structurally distinct from the typical four chain quaternary structure having two heavy and two light chains, for antibodies from other animals. See PCT / EP93 / 02214 (WO 94 / 04678 published 3 March 1994). Such IgG subtypes of the llamas lack the light chains and the CHI domain and are called heavy chain antibodies. These naturally occurring camelid antibodies consisting of only a heavy chain are functional and stable in the absence of light chain. The antigen-binding site of these heavy chain antibodies is formed only by a single domain, referred to as "VHH" (Kbnning et al., 2017) or, used synonymously herein, "VHH antibody domain".

[0152] Each light and heavy chain of an antibody, contains constant domains and variable domains. References to "VH" or "VH" refer to the variable region of an immunoglobulin heavy chain, including that of an antibody fragment. References to "VL" or "VL" refer to the variable region of an immunoglobulin light chain, such as in a primate antibody. The variable domain of a heavy chain antibody is called VHH. The VHH is composed of only one polypeptide chain of 15 kDa and is considered the smallest known natural domain with full antigen-binding capacity. Light and heavy chain variable domains contain a "framework" region interrupted by three hypervariable regions, also called "complementarity determining regions" or "CDRs" (see, e.g., Kabat et al., Sequences of Proteins of Immunological Interest, U.S. Department of Health and Human Services, 1991). The sequences of the framework regions of different light or heavy chains are relatively conserved within a species. The framework region of an antibody, that is the combined framework regions of the constituent light and heavy chains, serves to position and align the CDRs in three-dimensional space. The CDRs are primarily responsible for antigen binding.

[0153] The CDRs are typically referred to as CDR1, CDR2, and CDR3 (from the N-terminus to C- terminus), and are also typically identified by the chain in which the particular CDR is located. Thus, a VH CDR3 is located in the variable domain of the heavy chain of the antibody in which it is found, whereas a VL CDR1 is the CDR1 from the variable domain of the light chain of the antibody in which it is found. Light chain CDRs are sometimes referred to as CDR LI, CDR L2, and CDR L3. Heavy chain CDRs are sometimes referred to as CDR Hl, CDR H2, and CDR H3. VHH monoclonal antibodies have only a heavy chain, and thus include only one CDR1, CDR2 and CDR3. Generally, the CDR3 is primarily responsible for antigen specificity.

[0154] A VHH includes in an N- to C- direction, the following structural regions: N - FR1 - CDR1 - FR2 - CDR2 - FR3 - CDR3 - FR4 - C, wherein FR denotes a framework region amino acid sequence and CDR denotes a complementary determining region amino acid sequence (see, e.g., Kabat et al., Sequences of Proteins of Immunological Interest, U.S. Department of Health and Human Services, 1991).

[0155] As used in the present disclosure, a "VHH antibody domain" refers to the domain formed by the complete VHH, including (in an N- to C- direction) N - FR1 - CDR1 - FR2 - CDR2 - FR3 - CDR3 - FR4 - C. A VHH antibody domain can be part of a larger molecule to which the VHH antibody domain is covalently linked. Alternative, a VHH antibody domain can be a molecule of its own that is not covalently linked to any other molecular structure.

[0156] Since resulting paratopes are devoid of light chains, sdAbs afford the benefit of multiple reformatting options in a ‘plug-and-play’ manner involving beads-on-string assemblies or facile combinations with existing Fab-based paratopes for the generation of bispecifics (Lipinski et al., 2023; Chanier and Chames, 2019). The extent of the framework region and CDRs have been defined (see, Kabat et al., Sequences of Proteins of Immunological Interest, U.S. Department of Health and Human Services, 1991). The CDRs of the heavy chain variable domain are located at residues 31-35 (CDR-H1), residues 50-65 (CDR-H2) and residues 95-102 (CDR-H3) according to the Kabat numbering system. In antibodies (such as primate antibodies) that include a light chain, such as a primate antibody, the CDRs of the light chain variable domain are located at residues 24-34 (CDR-L1), residues 50-56 (CDR-L2) and residues 89-97 (CDR-L3) according to the Kabat numbering system. The Kabat database is now maintained online. The location of camelid CDRs can also be determined (see, for example, Sircar et al., J. Immunol. 186: 6357-6367, 2011); a program to determine camelid antibody structure, the Rosetta Antibody program, is available on the internet.

[0157] A "monoclonal antibody" is an antibody produced by a single clone of B-lymphocytes or by a cell into which the heavy chain gene (and optionally a light chain gene, such as for a primate antibody) of a single antibody have been transfected. Monoclonal antibodies may be obtained using a variety of techniques known to those skilled in the art, including standard hybridoma technology (see e.g. Kohler and Milstein, Eur. J. Immunol. (1976), vol. 5, p. 511-519; Antibodies: A Laboratory Manual, 2nd edition (2014), editor Greenfield, Cold Spring Harbor Laboratory Press (USA); Immunobiology, 5th ed. (2001), editors Janeway et al., Garland Publishing (USA)) and e.g. expression from a eukaryotic host cell transfected with a DNA molecule coding for the homogeneous antibody or from a prokaryotic host cell transfected with a DNA molecule coding for the homogeneous antibody.

[0158] VHH antibody domains can be obtained by genetic engineering to yield a small protein having high affinity for a target, resulting in a low molecular weight antibody derived protein. See e.g. Sellmann et al., 2020; U.S. Patent No. 5,759,808, issued June 2, 1998; see also Dumoulin et al., (2003); Pleschberger et al., (2003); Cortez-Retamozo et al., (2002); and Lauwereys et al., (1998).

[0159] In some embodiments, the VHH molecules can be produced as recombinant monoclonal antibodies or antigen binding fragments in different expression platforms, avoiding the use of hybridomas and mice. VHH monoclonal antibodies can be humanized monoclonal antibodies. In some embodiments, monoclonal antibodies can be chimeric antibodies. Without being bound by theory, a VHH monoclonal antibody has a molecular weight approximately one-tenth that of a human IgG molecule, and the protein has a physical diameter of only a few nanometers.

[0160] One consequence of the small size is the ability of the VHH monoclonal antibody to bind to antigenic sites that are functionally invisible to larger antibody proteins, such that VHH monoclonal antibodies are useful as reagents to detect antigens that are otherwise cryptic using classical immunological techniques, and thus are of use as therapeutic agents. Thus, yet another consequence of small size is that a camelid VHH monoclonal antibody can inhibit as a result of binding to a specific site in a groove or narrow cleft of a target protein, and hence can serve in a capacity that more closely resembles the function of a classical low molecular weight drug than that of a classical antibody.

[0161] Without being bound by theory, low molecular weight and compact size further result in camelid VHH monoclonal antibodies being extremely thermostable, stable to extreme pH and to proteolytic digestion, and poorly antigenic. Further, these molecules can be fully expressed in prokaryotic cells such as E. coli and are expressed as fusion proteins with bacteriophage and are functional.

[0162] "Humanizing" an antib ody / antibody sequence, as used herein, refers to the process of "germlining" where a non-human (such as camelid, llama or synthetic) antibody sequence is adapted to be more similar to a human antibody sequence by replacing one or more individual amino acids with the corresponding amino acids of a human antibody sequence. Typically, as human antibody sequence will be selected that is particularly close (i.e. has a high degree of sequence homology) to the non-human sequence. Such a human antibody sequence can be identified e.g. by a BLAST search. The corresponding amino acids can then be identified by a pairwise sequence alignment between the selected human antibody sequence and the non- human antibody sequence to be humanized. After humanization, the humanized antibody still binds to the same antigen as the original non-human antibody before humanization. Humanized immunoglobulins can be constructed by means of genetic engineering. A VHH antibody domain is easily humanized based on the human VH domain, which has a sequence that is highly homologous to the sequence of the VHH antibody domain. Furthermore, a VHH sequence can be adapted to reduce sequence liabilities. This may comprise, e.g., replacing amino acids susceptible for glycosylation, deamination, oxidization or isomerization by exchanging the respective amino acid(s) in the VHH sequence with the corresponding amino acid in the closest human germline sequence or with another amino acid (e.g. alanine). Similarly, the hydrophobic patches at the cell surface can be reduced by exchanging the respective amino acid(s) with less hydrophobic amino acid(s). Again, the amino acid replacements are chosen such that after the adaption the VHH sequence still binds to the same antigen and with similar characteristics as the VHH sequence before adaption.

[0163] VHHs can be used as modular building blocks for generating multivalent and / or multispecific antibody constructs, whereby "multivalent" means that the construct encompasses more than one single domain antibody and "multispecific" means that it encompasses single domain antibodies of more than one binding specificity.

[0164] At some occasions, the present disclosure states that a certain protein / amino acid sequence A is a "fragment" of another protein / amino acid sequence B. This means that, compared to protein / amino acid B, the protein / amino acid sequence A lacks one or more amino acids at the N-terminus and / or one or more amino acids at the C-terminus. Whether a protein / amino acid sequence lacks, compared to another protein / amino acid sequence, one or more amino acids at the N-terminus and / or one or more amino acids at the C-terminus can for example readily be determined upon forming a sequence alignment e.g. with the BLAST family of programs.

[0165] As the skilled person understands, when the present disclosure refers to an "VHH antibody domain or a fragment thereof', said fragment is an antigen-binding fragment. Thus, said fragment binds to the same antigen as the "full-length" VHH antibody domain according to the present disclosure from which said fragment is derived (i.e. to IL-18Ra resp. IL-18RP). Preferably, said fragment of said VHH antibody domain is a C-terminal fragment. This means that compared to the "complete" VHH antibody domain sequence said fragment lacks amino acids at the N-terminus.

[0166] When the present disclosure indicates that a VHH antibody domain or fragment thereof "of one VHH selected from the group consisting of VHH1, VHH2 and VHH3 as shown in the Table of CDRs" (or a corresponding wording), this means that said VHH antibody domain comprises the combination of CDRs or either VHH1 or VHH2 or VHH3, but not a mixture of CDRs selected from different of the listed VHHs. Thus, said VHH antibody domain or fragment thereof includes e.g. the combination of CDR1, CDR2 and CDR3 of VHH1 (SEQ ID NO: 23, 24 and 25), the combination of CDR1, CDR2 and CDR3 of VHH2 (SEQ ID NO: 26, 27, and 28), or the combination of CDR1, CDR2 and CDR3 of VHH3 (SEQ ID NO: 29, 30, and 31) etc., but not the combination of CDR1 and CDR2 of VHH1 and CDR3 of VHH2 (SEQ ID NO: 23, 24 and 28).

[0167] When the present disclosure defines that a VHH antibody domain or fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 as defined in e.g. (a) with modification, wherein the modification is e.g. that the sequence of at least one of CDR1, CDR2 and CDR3 is humanized, the skilled person is aware that this humanization exists compared to the corresponding sequence in the Table of CDRs that provides the combinations of CDR sequences without modification.

[0168] If the present disclosure indicates the existence of a modification which is "replacement, addition or deletion" of a certain number of amino acids (e.g. up to three), the skilled person understands that this is an individual replacement, addition or deletion of the indicated number of amino acids. Thus, the replaced, added or deleted amino acids may be at neighboring positions or at independent, isolated positions within the amino acid sequence. Moreover, as above, the skilled person is aware that this definition indicates the replacement, addition or deletion compared to the unmodified sequence in the Table of CDRs.

[0169] Embodiment 2: The VHH antibody domain or fragment thereof according to embodiment 1, wherein the modification in (b) is that the sequence of CDR1 and / or CDR2, but not the sequence of CDR3 is humanized.

[0170] Embodiment 3: The VHH antibody domain or fragment thereof according to any one of embodiments 1 or 2, wherein the modification in (b) is that the sequence of CDR1 is humanized, but not the sequence of CDR2 and CDR3.

[0171] Embodiment 4: The VHH antibody domain or fragment thereof according to any one of embodiments 1 or 2, wherein the modification in (b) is that the sequence of CDR2 is humanized, but not the sequence of CDR1 and CDR3. Embodiment 5: The VHH antibody domain or fragment thereof according to any one of embodiments 1 to 4, wherein the modification in (b) is that the sequence of one, but not more than one of CDR1, CDR2 and CDR3 is humanized.

[0172] Embodiment 6: The VHH antibody domain or fragment thereof according to any one of embodiments 1 to 5, wherein said humanization of said CDR(s) is by replacing at least one amino acid in the sequence of said CDR by the corresponding amino acid of a human VH domain.

[0173] Embodiment 7: The VHH antibody domain or fragment thereof according to any one of embodiments 1 to 6, wherein said humanization of said CDR(s) is by replacing up to three amino acids in the sequence of said CDR by the corresponding amino acid of a human VH domain.

[0174] Embodiment 8: The VHH antibody domain or fragment thereof according to any one of embodiments 1 to 6, wherein said humanization of said CDR(s) is by replacing up to three amino acids in the sequence of CDR1 and / or CDR2 and up to one amino acid in the sequence of CDR3 by the corresponding amino acid of a human VH domain.

[0175] Embodiment 9: The VHH antibody domain or fragment thereof according to any one of embodiments 1 to 6, wherein said humanization of said CDR(s) is by replacing up to two amino acids in the sequence of said CDR by the corresponding amino acid of a human VH domain.

[0176] Embodiment 10: The VHH antibody domain or fragment thereof according to any one of embodiments 1 to 6, wherein said humanization of said CDR(s) is by replacing up to two amino acids in the sequence of CDR1 and / or CDR2 and up to one amino acid in the sequence of CDR3 by the corresponding amino acid of a human VH domain.

[0177] Embodiment 11 : The VHH antibody domain or fragment thereof according to any one of embodiments 1 to 10, wherein said humanization of said CDR(s) is by replacing one amino acid in the sequence of said CDR by the corresponding amino acid of a human VH domain.

[0178] Embodiment 12: The VHH antibody domain or fragment thereof according to any one of embodiments 1 to 11, wherein the modification in (c) is - the replacement, addition or deletion of up to three amino acids in CDR1,

[0179] - the replacement, addition or deletion of up to three amino acids in CDR2 and / or

[0180] - the replacement, addition or deletion of up to one amino acid in CDR3.

[0181] Embodiment 13: The VHH antibody domain or fragment thereof according to any one of embodiments 1 to 11, wherein the modification in (c) is

[0182] - the replacement, addition or deletion of up to two amino acids in CDR1,

[0183] - the replacement, addition or deletion of up to two amino acids in CDR2 and / or

[0184] - the replacement, addition or deletion of up to two amino acids in CDR3;

[0185] Embodiment 14: The VHH antibody domain or fragment thereof according to any one of embodiments 1 to 11, wherein the modification in (c) is

[0186] - the replacement, addition or deletion of up to two amino acids in CDR1;

[0187] - the replacement, addition or deletion of up to two amino acids in CDR2; and / or

[0188] - the replacement, addition or deletion of up to one amino acid in CDR3.

[0189] Embodiment 15: The VHH antibody domain or fragment thereof according to any one of embodiments 1 to 11, wherein the modification in (c) is

[0190] - the replacement, addition or deletion of up to two amino acids in CDR1 and / or

[0191] - the replacement, addition or deletion of up to two amino acids in CDR2; wherein the sequence of CDR3 is unmodified.

[0192] As a skilled person understands, the indication that the "sequence of CDR3 is unmodified" means that the sequence is unmodified compared to the sequence provided for CDR3 for the VHH at issue in the Table of CDRs.

[0193] Embodiment 16: The VHH antibody domain or fragment thereof according to any one of embodiments 1 to 11, wherein the modification in (c) is

[0194] - the replacement, addition or deletion of up to two amino acids in CDR1, wherein the sequence of CDR2 and CDR3 is unmodified.

[0195] Embodiment 17: The VHH antibody domain or fragment thereof according to any one of embodiments 1 to 11, wherein the modification in (c) is

[0196] - the replacement, addition or deletion of up to two amino acids in CDR2, wherein the sequence of CDR1 and CDR3 is unmodified.

[0197] Embodiment 18: The VHH antibody domain or fragment thereof according to any one of embodiments 1 to 11, wherein the modification in (c) is

[0198] - the replacement, addition or deletion of up to one amino acid in CDR1;

[0199] - the replacement, addition or deletion of up to one amino acid in CDR2; and / or

[0200] - the replacement, addition or deletion of up to one amino acid in CDR3.

[0201] Embodiment 19: The VHH antibody domain or fragment thereof according to any one of embodiments 1 to 11, wherein the modification in (c) is

[0202] - the replacement, addition or deletion of one amino acid in CDR1 and / or

[0203] - the replacement, addition or deletion of one amino acid in CDR2; wherein the sequence of CDR3 is unmodified.

[0204] Embodiment 20: The VHH antibody domain or fragment thereof according to any one of embodiments 1 to 11, wherein the modification in (c) is

[0205] - the replacement, addition or deletion of one amino acid in CDR1, wherein the sequence of CDR2 and CDR3 is unmodified.

[0206] Embodiment 21 : The VHH antibody domain or fragment thereof according to any one of embodiments 1 to 11, wherein the modification in (c) is

[0207] - the replacement, addition or deletion of one amino acid in CDR2, wherein the sequence of CDR1 and CDR3 is unmodified.

[0208] Embodiment 22: The VHH antibody domain or fragment thereof according to any one of embodiments 1 to 21, wherein the modification in (c) comprises only the replacement, but not the addition or deletion of amino acids.

[0209] Embodiment 23: The VHH antibody domain or fragment thereof according to any one of embodiments 1 to 22, wherein said replacement is a conservative amino acid replacement.

[0210] As used herein, a "conservative amino acid replacement" refers to the replacement of an amino acid by another, biologically similar amino acid. Conservative replacements are not likely to change the shape or characteristics of a protein / amino acid sequence. Examples of conservative replacements include the replacement of one hydrophobic residue such as isoleucine, valine, leucine or methionine for another, or the substitution of one polar residue for another, such as the substitution of arginine for lysine, glutamic for aspartic acid, or glutamine for asparagine.

[0211] Second aspect of the present disclosure (also referred to as "Embodiment 24"): According to a second aspect, the present disclosure relates to a VHH antibody domain or a fragment thereof, wherein

[0212] (A) said VHH antibody domain comprises the amino acid sequence of a VHH selected from the group consisting of VHH1, VHH2, VHH3, VHH4, VHH5, VHH6, VHH8, VHH9, VHH10 and VHH11 as shown in the Table of VHH Sequences;

[0213] (B) said VHH antibody domain comprises a VHH sequence as defined in (A) with modification, wherein the modification is that said sequence is humanized;

[0214] (C) said VHH antibody domain comprises a VHH sequence as defined in (A) with modification, wherein the modification is the replacement, addition or deletion of up to 25 amino acids; or

[0215] (D) said VHH antibody domain comprises a VHH sequence that is at least 75% identical to a VHH sequence referred to in (A);

[0216] Table of VHH Sequences:

[0217] Third aspect of the present disclosure (also referred to as "Embodiment 25"): According to a third aspect, the present disclosure relates to a VHH antibody domain or a fragment thereof, wherein

[0218] (A) said VHH antibody domain consists of the amino acid sequence of a VHH selected from the group consisting of VHH1, VHH2, VHH3, VHH4, VHH5, VHH6, VHH8, VHH9, VHH10 and VHH11 as shown in the Table of VHH Sequences;

[0219] (B) said VHH antibody domain consists of a VHH sequence as defined in (A) with modification, wherein the modification is that said sequence is humanized;

[0220] (C) said VHH antibody domain consists of a VHH sequence as defined in (A) with modification, wherein the modification is the replacement, addition or deletion of up to 25 amino acids; or

[0221] (D) said VHH antibody domain consists of a VHH sequence that is at least 75% identical to a VHH sequence referred to in (A). To the second and third aspects of the present disclosure and the embodiments referring thereto, the same explanations and definitions apply accordingly as for the first aspect of the present disclosure and the embodiments referring thereto.

[0222] As the skilled person is aware, if the present disclosure indicates that a VHH antibody domain comprises e.g. "the VHH sequence of a VHH selected from the group consisting of VHH1, VHH2, VHH3, VHH4, VHH5, VHH6, VHH8, VHH9, VHH10 and VHH11 as shown in the Table of VHH Sequences", this means that said VHH antibody domain comprises one and (not multiple or all) of the sequences listed in the Table of VHH Sequences.

[0223] Thus, if the present disclosure indicates that a VHH antibody domain or fragment thereof comprises / consists e.g. of a VHH sequence as defined in (A) "with modification, wherein the modification is that said sequence is humanized", the skilled person is aware that this means that this humanization exists compared to the corresponding sequence in the Table of VHH Sequences that provides the sequences without modification.

[0224] If the present disclosure states that a certain sequence A "is at least x % identical" to another sequence B, this is synonymous to the statement that sequence A "has x % identity" to sequence B. The statement reflects a relationship between the two polypeptide sequences A and B determined by comparing the sequences. In general, identity refers to an exact amino acid to amino acid correspondence of the two polypeptide sequences, respectively, over the length of the sequences being compared. For sequences where there is not an exact correspondence, a percentage to which the two sequences are identical may be determined. In general, the two sequences to be compared are aligned to give a maximum correlation between the sequences. This may include inserting "gaps" in either one or both sequences, to enhance the degree of alignment. A % identity may be determined over the whole length of each of the sequences being compared (so-called global alignment), that is particularly suitable for sequences of the same or very similar length, or over shorter, defined lengths (so-called local alignment), that is more suitable for sequences of unequal length.

[0225] Methods for comparing the identity of two or more sequences are well known in the art. Thus, for instance, programs available in the Wisconsin Sequence Analysis Package, version 9.1 (Devereux J et al., 1984), for example the programs BESTFIT and GAP, may be used to determine the % identity between two polynucleotides and the % identity between two polypeptide sequences. BESTFIT uses the "local homology" algorithm of Smith and Waterman (1981) and finds the best single region of similarity between two sequences. Other programs for determining identity sequences are also known in the art, for instance the BLAST family of programs (Altschul S F et al, 1990, Altschul S F et al, 1997, accessible through the home page of the NCBI at www.ncbi.nlm.nih.gov) and FASTA (Pearson WR, 1990). Preferably, % identity according to the present disclosure is determined according to the BLAST family of programs (Altschul S F et al, 1990, Altschul S F et al, 1997, accessible through the home page of the NCBI at www.ncbi.nlm.nih.gov).

[0226] Embodiment 26: The VHH antibody domain or fragment thereof according to any one of embodiments 24 or 25, wherein said fragment of said VHH antibody domain comprises at least 75% of the amino acids of the sequence of said VHH antibody domain. As the skilled person understands, this means that that fragment lacks up to a quarter of the total number of amino acids of said VHH antibody domain, wherein, compared to the "complete" VHH antibody domain sequence said amino acids are lacking either at the N-terminus or at the C-terminus.

[0227] Embodiment 27: The VHH antibody domain or fragment thereof according to any one of embodiments 24 or 25, wherein said fragment of said VHH antibody domain comprises at least 80% of the amino acids of the sequence of said VHH antibody domain.

[0228] Embodiment 28: The VHH antibody domain or fragment thereof according to any one of embodiments 24 or 25, wherein said fragment of said VHH antibody domain comprises at least 85% of the amino acids of the sequence of said VHH antibody domain.

[0229] Embodiment 29: The VHH antibody domain or fragment thereof according to any one of embodiments 24 or 25, wherein said fragment of said VHH antibody domain comprises at least 90% of the amino acids of the sequence of said VHH antibody domain.

[0230] Embodiment 30: The VHH antibody domain or fragment thereof according to any one of embodiments 24 or 25, wherein said fragment of said VHH antibody domain comprises at least 95% of the amino acids of the sequence of said VHH antibody domain. 973 Embodiment 31 : The VHH antibody domain or fragment thereof according to any one of

[0231] 974 embodiments 24 or 25, wherein said fragment of said VHH antibody domain comprises at least

[0232] 975 98% of the amino acids of the sequence of said VHH antibody domain.

[0233] 976

[0234] 977 Embodiment 32: The VHH antibody domain or fragment thereof according to any one of

[0235] 978 embodiments 24 or 25, wherein said fragment of said VHH antibody domain comprises at least

[0236] 979 99% of the amino acids of the sequence of said VHH antibody domain.

[0237] 980

[0238] 981 Embodiment 33: The VHH antibody domain or fragment thereof according to any one of

[0239] 982 embodiments 24 to 32, wherein said fragment of said VHH antibody domain comprises the

[0240] 983 complementarity determining regions CDR1, CDR2 and CDR3.

[0241] 984

[0242] 985 Embodiment 34: The VHH antibody domain or fragment thereof according to any one of

[0243] 986 embodiments 24 to 33, wherein said fragment of said VHH antibody domain comprises at least

[0244] 987 the sequence from the N-terminus of CDR1 to the C-terminus of CDR3 of said VHH antibody

[0245] 988 domain.

[0246] 989

[0247] 990 Embodiment 35: The VHH antibody domain or fragment thereof according to any one of

[0248] 991 embodiments 24 to 34, wherein in (A) said fragment of said VHH antibody domain comprises

[0249] 992 all the complementarity determining regions (CDRs) of said VHH antibody domain.

[0250] 993

[0251] 994 Embodiment 36: The VHH antibody domain or fragment thereof according to any one of

[0252] 995 embodiments 24 to 35, wherein in (B) said humanization of said sequence is by replacing at

[0253] 996 least one amino acid of said sequence by the corresponding amino acid of a human VH (variable

[0254] 997 heavy) domain.

[0255] 998

[0256] 999 Embodiment 37: The VHH antibody domain or fragment thereof according to any one of

[0257] 1000 embodiments 24 to 36, wherein in (B) said humanization of said sequence is by (individually)

[0258] 1001 replacing up to 25 amino acids of said sequence by the corresponding amino acids of a human

[0259] 1002 VH domain.

[0260] 1003

[0261] 1004 As the skilled person understands, if the present disclosure refers to the replacement of an amino

[0262] 1005 acid of a certain sequence / domain A by the "corresponding" amino acid of a certain

[0263] 1006 sequence / domain B, this designates that said amino acid of sequence / domain A is replaced by 1007 the amino acid in sequence / domain B that in an alignment of the two sequences aligns with said

[0264] 1008 amino acid of sequence / domain A.

[0265] 1009

[0266] 1010 Embodiment 38: The VHH antibody domain or fragment thereof according to any one of

[0267] 1011 embodiments 24 to 36, wherein in (B) said humanization of said sequence is by replacing up to

[0268] 1012 20 amino acids of said sequence by the corresponding amino acids of a human VH domain.

[0269] 1013

[0270] 1014 Embodiment 39: The VHH antibody domain or fragment thereof according to any one of

[0271] 1015 embodiments 24 to 36, wherein in (B) said humanization of said sequence is by replacing up to

[0272] 1016 15 amino acids of said sequence by the corresponding amino acids of a human VH domain.

[0273] 1017

[0274] 1018 Embodiment 40: The VHH antibody domain or fragment thereof according to any one of

[0275] 1019 embodiments 24 to 36, wherein in (B) said humanization of said sequence is by replacing up to

[0276] 1020 10 amino acids of said sequence by the corresponding amino acids of a human VH domain.

[0277] 1021

[0278] 1022 Embodiment 41 : The VHH antibody domain or fragment thereof according to any one of

[0279] 1023 embodiments 24 to 36, wherein in (B) said humanization of said sequence is by replacing up to

[0280] 1024 5 amino acids of said sequence by the corresponding amino acids of a human VH domain.

[0281] 1025

[0282] 1026 Embodiment 42: The VHH antibody domain or fragment thereof according to any one of

[0283] 1027 embodiments 24 to 36, wherein in (B) said humanization of said sequence is by replacing up to

[0284] 1028 3 amino acids of said sequence by the corresponding amino acids of a human VH domain.

[0285] 1029

[0286] 1030 Embodiment 43: The VHH antibody domain or fragment thereof according to any one of

[0287] 1031 embodiments 24 to 36, wherein in (B) said humanization of said sequence is by replacing up to

[0288] 1032 2 amino acids of said sequence by the corresponding amino acids of a human VH domain.

[0289] 1033

[0290] 1034 Embodiment 44: The VHH antibody domain or fragment thereof according to any one of

[0291] 1035 embodiments 24 to 36, wherein in (B) said humanization of said sequence is by replacing one

[0292] 1036 amino acid of said sequence by the corresponding amino acid of a human VH domain.

[0293] 1037

[0294] 1038 Embodiment 45: The VHH antibody domain or fragment thereof according to any one of

[0295] 1039 embodiments 24 to 44, wherein in (B) said humanization is within the framework regions of

[0296] 1040 said VHH antibody domain and / or within the CDRs of said VHH antibody domain. 1041

[0297] 1042 Embodiment 46: The VHH antibody domain or fragment thereof according to any one of

[0298] 1043 embodiments 24 to 44, wherein in (B) said humanization is within the framework regions of

[0299] 1044 said VHH antibody domain, but not within the CDRs of said VHH antibody domain.

[0300] 1045

[0301] 1046 Embodiment 47: The VHH antibody domain or fragment thereof according to any one of

[0302] 1047 embodiments 24 to 36, wherein in (B) said humanization is within the CDRs of said VHH

[0303] 1048 antibody domain, but not within the framework regions of said VHH antibody domain.

[0304] 1049

[0305] 1050 Embodiment 48: The VHH antibody domain or fragment thereof according to any one of

[0306] 1051 embodiments 24 to 45 or 47, wherein in (B) said humanization within the CDRs of said VHH

[0307] 1052 antibody domain is within CDR1, CDR2 and / or CDR3.

[0308] 1053

[0309] 1054 Embodiment 49: The VHH antibody domain or fragment thereof according to any one of

[0310] 1055 embodiments 24 to 45 or 47 to 48, wherein in (B) said humanization within the CDRs of said

[0311] 1056 VHH antibody domain is within CDR1 and / or CDR2.

[0312] 1057

[0313] 1058 Embodiment 50: The VHH antibody domain or fragment thereof according to any one of

[0314] 1059 embodiments 24 to 45 or 47 to 49, wherein in (B) said humanization within the CDRs of said

[0315] 1060 VHH antibody domain is within CDR1.

[0316] 1061

[0317] 1062 Embodiment 51: The VHH antibody domain or fragment thereof according to any one of

[0318] 1063 embodiments 24 to 45 or 47 to 50, wherein in (B) said humanization within the CDRs of said

[0319] 1064 VHH antibody domain is within CDR2.

[0320] 1065

[0321] 1066 Embodiment 52: The VHH antibody domain or fragment thereof according to any one of

[0322] 1067 embodiments 24 to 51, wherein in (B) said humanization within the CDRs of said VHH

[0323] 1068 antibody domain is not within CDR3.

[0324] 1069

[0325] 1070 Embodiment 53: The VHH antibody domain or fragment thereof according to any one of

[0326] 1071 embodiments 24 to 52, wherein in (C) the modification is the replacement, addition or deletion

[0327] 1072 of up to 20 amino acids.

[0328] 1073 1074 Embodiment 54: The VHH antibody domain or fragment thereof according to any one of

[0329] 1075 embodiments 24 to 52, wherein in (C) the modification is the replacement, addition or deletion

[0330] 1076 of up to 15 amino acids.

[0331] 1077

[0332] 1078 Embodiment 55: The VHH antibody domain or fragment thereof according to any one of

[0333] 1079 embodiments 24 to 52, wherein in (C) the modification is the replacement, addition or deletion

[0334] 1080 of up to 10 amino acids.

[0335] 1081

[0336] 1082 Embodiment 56: The VHH antibody domain or fragment thereof according to any one of

[0337] 1083 embodiments 24 to 52, wherein in (C) the modification is the replacement, addition or deletion

[0338] 1084 of up to 5 amino acids.

[0339] 1085

[0340] 1086 Embodiment 57: The VHH antibody domain or fragment thereof according to any one of

[0341] 1087 embodiments 24 to 52, wherein in (C) the modification is the replacement, addition or deletion

[0342] 1088 of up to 3 amino acids.

[0343] 1089

[0344] 1090 Embodiment 58: The VHH antibody domain or fragment thereof according to any one of

[0345] 1091 embodiments 24 to 52, wherein in (C) the modification is the replacement, addition or deletion

[0346] 1092 of up to 2 amino acids.

[0347] 1093

[0348] 1094 Embodiment 59: The VHH antibody domain or fragment thereof according to any one of

[0349] 1095 embodiments 24 to 52, wherein in (C) the modification is the replacement, addition or deletion

[0350] 1096 of one amino acid.

[0351] 1097

[0352] 1098 Embodiment 60: The VHH antibody domain or fragment thereof according to any one of

[0353] 1099 embodiments 24 to 59, wherein the modification in (C) comprises only the replacement, but not

[0354] 1100 the addition or deletion of amino acids.

[0355] 1101

[0356] 1102 Embodiment 61 : The VHH antibody domain or fragment thereof according to any one of

[0357] 1103 embodiments 24 to 60, wherein in (A) said VHH antibody domain or fragment thereof

[0358] 1104 comprises the complementarity determining regions CDR1, CDR2 and CDR3 of one VHH

[0359] 1105 selected from the group consisting of VHH1, VHH2, VHH3, VHH4, VHH5, VHH6, VHH8,

[0360] 1106 VHH9, VHH10 and VHH11 as shown in the Table of CDRs.

[0361] 1107 1108 Embodiment 62: The VHH antibody domain or fragment thereof according to any one of

[0362] 1109 embodiments 24 to 61, wherein in (B) to (D)

[0363] 1110 (a) said VHH antibody domain or fragment thereof comprises the complementarity

[0364] 1111 determining regions CDR1, CDR2 and CDR3 of one VHH selected from the group

[0365] 1112 consisting of VHH1, VHH2, VHH3, VHH4, VHH5, VHH6, VHH8, VHH9, VHH10

[0366] 1113 and VHH11 as shown in the Table of CDRs;

[0367] 1114 (b) said VHH antibody domain or fragment thereof comprises the complementarity

[0368] 1115 determining regions CDR1, CDR2 and CDR3 as defined in (a) with modification,

[0369] 1116 wherein the modification is that the sequence of at least one of CDR1, CDR2 and

[0370] 1117 CDR3 is humanized;

[0371] 1118 (c) said VHH antibody domain or fragment thereof comprises the complementarity

[0372] 1119 determining regions CDR1, CDR2 and CDR3 as defined in (a) with modification,

[0373] 1120 wherein the modification is

[0374] 1121 - the replacement, addition or deletion of up to three amino acids in CDR1,

[0375] 1122 - the replacement, addition or deletion of up to three amino acids in CDR2

[0376] 1123 and / or

[0377] 1124 - the replacement, addition or deletion of up to three amino acids in CDR3.

[0378] 1125

[0379] 1126 Embodiment 63 : The VHH antibody domain or fragment thereof according to embodiment 62,

[0380] 1127 wherein the modification in (b) is that the sequence of CDR1 and / or CDR2, but not the sequence

[0381] 1128 of CDR3 is humanized.

[0382] 1129

[0383] 1130 Embodiment 64: The VHH antibody domain or fragment thereof according to any one of

[0384] 1131 embodiments 62 or 63, wherein the modification in (b) is that the sequence of CDR1 is

[0385] 1132 humanized, but not the sequence of CDR2 and CDR3.

[0386] 1133

[0387] 1134 Embodiment 65: The VHH antibody domain or fragment thereof according to any one of

[0388] 1135 embodiments 62 or 63, wherein the modification in (b) is that the sequence of CDR2 is

[0389] 1136 humanized, but not the sequence of CDR1 and CDR3.

[0390] 1137

[0391] 1138 Embodiment 66: The VHH antibody domain or fragment thereof according to any one of

[0392] 1139 embodiments 62 to 65, wherein the modification in (b) is that the sequence of one, but not more

[0393] 1140 than one of CDR1, CDR2 and CDR3 is humanized.

[0394] 1141 1142 Embodiment 67: The VHH antibody domain or fragment thereof according to any one of

[0395] 1143 embodiments 62 to 66, wherein said humanization of said CDR(s) is by replacing at least one

[0396] 1144 amino acid in the sequence of said CDR by the corresponding amino acid of a human VH

[0397] 1145 domain.

[0398] 1146

[0399] 1147 Embodiment 68: The VHH antibody domain or fragment thereof according to any one of

[0400] 1148 embodiments 62 to 67, wherein said humanization of said CDR(s) is by replacing up to three

[0401] 1149 amino acids in the sequence of said CDR by the corresponding amino acid of a human VH

[0402] 1150 domain.

[0403] 1151

[0404] 1152 Embodiment 69: The VHH antibody domain or fragment thereof according to any one of

[0405] 1153 embodiments 62 to 67, wherein said humanization of said CDR(s) is by replacing up to three

[0406] 1154 amino acids in the sequence of CDR1 and / or CDR2 and up to one amino acid in the sequence

[0407] 1155 of CDR3 by the corresponding amino acid of a human VH domain.

[0408] 1156

[0409] 1157 Embodiment 70: The VHH antibody domain or fragment thereof according to any one of

[0410] 1158 embodiments 62 to 67, wherein said humanization of said CDR(s) is by replacing up to two

[0411] 1159 amino acids in the sequence of said CDR by the corresponding amino acid of a human VH

[0412] 1160 domain.

[0413] 1161

[0414] 1162 Embodiment 71 : The VHH antibody domain or fragment thereof according to any one of

[0415] 1163 embodiments 62 to 67, wherein said humanization of said CDR(s) is by replacing up to two

[0416] 1164 amino acids in the sequence of CDR1 and / or CDR2 and up to one amino acid in the sequence

[0417] 1165 of CDR3 by the corresponding amino acid of a human VH domain.

[0418] 1166

[0419] 1167 Embodiment 72: The VHH antibody domain or fragment thereof according to any one of

[0420] 1168 embodiments 62 to 71, wherein said humanization of said CDR(s) is by replacing one amino

[0421] 1169 acid in the sequence of said CDR by the corresponding amino acid of a human VH domain.

[0422] 1170

[0423] 1171 Embodiment 73: The VHH antibody domain or fragment thereof according to any one of

[0424] 1172 embodiments 62 to 72, wherein the modification in (c) is

[0425] 1173 - the replacement, addition or deletion of up to three amino acids in CDR1,

[0426] 1174 - the replacement, addition or deletion of up to three amino acids in CDR2 and / or

[0427] 1175 - the replacement, addition or deletion of up to two amino acids in CDR3. 1176

[0428] 1177 Embodiment 74: The VHH antibody domain or fragment thereof according to any one of

[0429] 1178 embodiments 62 to 72, wherein the modification in (c) is

[0430] 1179 - the replacement, addition or deletion of up to three amino acids in CDR1,

[0431] 1180 - the replacement, addition or deletion of up to three amino acids in CDR2 and / or

[0432] 1181 - the replacement, addition or deletion of up to one amino acid in CDR3.

[0433] 1182

[0434] 1183 Embodiment 75: The VHH antibody domain or fragment thereof according to any one of

[0435] 1184 embodiments 62 to 72, wherein the modification in (c) is

[0436] 1185 - the replacement, addition or deletion of up to three amino acids in CDR1, and / or

[0437] 1186 - the replacement, addition or deletion of up to three amino acids in CDR2,

[0438] 1187 wherein the sequence of CDR3 is unmodified.

[0439] 1188

[0440] 1189 Embodiment 76: The VHH antibody domain or fragment thereof according to any one of

[0441] 1190 embodiments 62 to 72, wherein the modification in (c) is

[0442] 1191 - the replacement, addition or deletion of up to two amino acids in CDR1,

[0443] 1192 - the replacement, addition or deletion of up to two amino acids in CDR2 and / or

[0444] 1193 - the replacement, addition or deletion of up to two amino acids in CDR3;

[0445] 1194

[0446] 1195 Embodiment 77: The VHH antibody domain or fragment thereof according to any one of

[0447] 1196 embodiments 62 to 72, wherein the modification in (c) is

[0448] 1197 - the replacement, addition or deletion of up to two amino acids in CDR1;

[0449] 1198 - the replacement, addition or deletion of up to two amino acids in CDR2; and / or

[0450] 1199 - the replacement, addition or deletion of up to one amino acid in CDR3.

[0451] 1200

[0452] 1201 Embodiment 78: The VHH antibody domain or fragment thereof according to any one of

[0453] 1202 embodiments 62 to 72, wherein the modification in (c) is

[0454] 1203 - the replacement, addition or deletion of up to two amino acids in CDR1 and / or

[0455] 1204 - the replacement, addition or deletion of up to two amino acids in CDR2;

[0456] 1205 wherein the sequence of CDR3 is unmodified.

[0457] 1206

[0458] 1207 As the skilled person is aware, "unmodified" means unmodified compared to the sequence in

[0459] 1208 the Table of CDRs.

[0460] 1209 1210 Embodiment 79: The VHH antibody domain or fragment thereof according to any one of

[0461] 1211 embodiments 62 to 72, wherein the modification in (c) is

[0462] 1212 - the replacement, addition or deletion of up to two amino acids in CDR1,

[0463] 1213 wherein the sequence of CDR2 and CDR3 is unmodified.

[0464] 1214

[0465] 1215 Embodiment 80: The VHH antibody domain or fragment thereof according to any one of

[0466] 1216 embodiments 62 to 72, wherein the modification in (c) is

[0467] 1217 - the replacement, addition or deletion of up to two amino acids in CDR2,

[0468] 1218 wherein the sequence of CDR1 and CDR3 is unmodified.

[0469] 1219

[0470] 1220 Embodiment 81 : The VHH antibody domain or fragment thereof according to any one of

[0471] 1221 embodiments 62 to 72, wherein the modification in (c) is

[0472] 1222 - the replacement, addition or deletion of up to one amino acid in CDR1;

[0473] 1223 - the replacement, addition or deletion of up to one amino acid in CDR2; and / or

[0474] 1224 - the replacement, addition or deletion of up to one amino acid in CDR3.

[0475] 1225

[0476] 1226 Embodiment 82: The VHH antibody domain or fragment thereof according to any one of

[0477] 1227 embodiments 62 to 72, wherein the modification in (c) is

[0478] 1228 - the replacement, addition or deletion of one amino acid in CDR1 and / or

[0479] 1229 - the replacement, addition or deletion of one amino acid in CDR2;

[0480] 1230 wherein the sequence of CDR3 is unmodified.

[0481] 1231

[0482] 1232 Embodiment 83: The VHH antibody domain or fragment thereof according to any one of

[0483] 1233 embodiments 62 to 72, wherein the modification in (c) is

[0484] 1234 - the replacement, addition or deletion of one amino acid in CDR1,

[0485] 1235 wherein the sequence of CDR2 and CDR3 is unmodified.

[0486] 1236

[0487] 1237 Embodiment 84: The VHH antibody domain or fragment thereof according to any one of

[0488] 1238 embodiments 62 to 72, wherein the modification in (c) is

[0489] 1239 - the replacement, addition or deletion of one amino acid in CDR2,

[0490] 1240 wherein the sequence of CDR1 and CDR3 is unmodified.

[0491] 1241 1242 Embodiment 85: The VHH antibody domain or fragment thereof according to any one of

[0492] 1243 embodiments 62 to 84, wherein the modification in (c) comprises only the replacement, but not

[0493] 1244 the addition or deletion of amino acids.

[0494] 1245

[0495] 1246 Embodiment 86: The VHH antibody domain or fragment thereof according to any one of

[0496] 1247 embodiments 62 to 85, wherein said replacement is a conservative amino acid replacement.

[0497] 1248

[0498] 1249 Embodiment 87: The VHH antibody domain or fragment thereof according to any one of

[0499] 1250 embodiments 1 to 86, wherein said fragment consists of at least 100 amino acids.

[0500] 1251

[0501] 1252 Embodiment 88: The VHH antibody domain or fragment thereof according to any one of

[0502] 1253 embodiments 1 to 86, wherein said fragment consists of at least 105 amino acids.

[0503] 1254

[0504] 1255 Embodiment 89: The VHH antibody domain or fragment thereof according to any one of

[0505] 1256 embodiments 1 to 86, wherein said fragment consists of at least 110 amino acids.

[0506] 1257

[0507] 1258 Embodiment 90: The VHH antibody domain or fragment thereof according to any one of

[0508] 1259 embodiments 1 to 86, wherein said fragment consists of at least 115 amino acids.

[0509] 1260

[0510] 1261 Embodiment 91 : The VHH antibody domain or fragment thereof according to any one of

[0511] 1262 embodiments 1 to 90, wherein said VHH antibody domain is an anti-IL-18Ra VHH antibody

[0512] 1263 domain.

[0513] 1264

[0514] 1265 Embodiment 92: The VHH antibody domain or fragment thereof according to any one of

[0515] 1266 embodiments 1 to 91, wherein said VHH antibody domain is specific for IL-18Ra.

[0516] 1267

[0517] 1268 Embodiment 93: The VHH antibody domain or fragment thereof according to any one of

[0518] 1269 embodiments 1 to 92, wherein said VHH antibody domain or fragment thereof binds to IL-

[0519] 1270 18Ra.

[0520] 1271

[0521] 1272 Embodiment 94: The VHH antibody domain or fragment thereof according to any one of

[0522] 1273 embodiments 1 to 93, wherein said VHH antibody domain or fragment thereof binds

[0523] 1274 specifically to IL-18Ra.

[0524] 1275 1276 Embodiment 95: The VHH antibody domain or fragment thereof according to any one of

[0525] 1277 embodiments 1 to 94, wherein said VHH antibody domain or fragment thereof does not bind to

[0526] 1278 IL-18RP.

[0527] 1279

[0528] 1280 Embodiment 96: The VHH antibody domain or fragment thereof according to any one of

[0529] 1281 embodiments 1 to 95, wherein said VHH antibody domain or fragment thereof is capable of

[0530] 1282 specifically binding to IL-18Ra ECD.

[0531] 1283

[0532] 1284 In some embodiments, if the present disclosure states that a first molecule / molecular group (e.g.

[0533] 1285 an antibody / antibody component) "is capable of specifically binding" / " specifically binds" to a

[0534] 1286 second molecule / molecular group (e.g. an antigen of interest), this means that the first

[0535] 1287 molecule / molecular group (in this example the antibody) binds to said second

[0536] 1288 molecule / molecular group (in this example the antigen of interest) with an affinity that is at

[0537] 1289 least ten-fold greater than its affinity for other molecules / molecular groups, in particular other

[0538] 1290 molecule / molecular group in the human body (in this example at least ten-fold greater than its

[0539] 1291 affinity for binding to non-specific antigens (e.g., BSA, casein) other than said antigen of

[0540] 1292 interest (or closely related antigens)). In a preferred embodiment, a first molecule / molecular

[0541] 1293 group (e.g. an antibody / antibody component) that "specifically binds" to a second

[0542] 1294 molecule / molecular group (e.g. an antigen of interest) binds to said antigen with an affinity that

[0543] 1295 is at least 100-fold greater than its affinity for other molecules / molecular groups, in particular

[0544] 1296 other molecule / molecular group in the human body (in this example at least 100-fold greater

[0545] 1297 than its affinity for binding to non-specific antigens other than said antigen of interest (or

[0546] 1298 closely related antigens)). Typically said binding will be determined under physiological

[0547] 1299 conditions. A first molecule / molecular group that "specifically binds" to a second

[0548] 1300 molecule / molecular group may bind to that second molecule / molecular group with a KD of

[0549] 1301 1 * 10'7M or stronger.

[0550] 1302

[0551] 1303 Embodiment 97: The VHH antibody domain or fragment thereof according to any one of

[0552] 1304 embodiments 1 to 96, wherein said VHH antibody domain or fragment thereof binds to

[0553] 1305 recombinant human IL-18Ra ECD with a KD of 1x1 O'6M or stronger.

[0554] 1306

[0555] 1307 Embodiment 98: The VHH antibody domain or fragment thereof according to any one of

[0556] 1308 embodiments 1 to 97, wherein said VHH antibody domain or fragment thereof binds to

[0557] 1309 recombinant human IL-18Ra ECD with a KD of 1x1 O'7M or stronger. 1310

[0558] 1311 Embodiment 99: The VHH antibody domain or fragment thereof according to any one of

[0559] 1312 embodiments 1 to 97, wherein said VHH antibody domain or fragment thereof binds to

[0560] 1313 recombinant human IL-18Ra ECD with a KD of 5x1 O'8M or stronger.

[0561] 1314

[0562] 1315 Embodiment 100: The VHH antibody domain or fragment thereof according to any one of

[0563] 1316 embodiments 1 to 97, wherein said VHH antibody domain or fragment thereof binds to

[0564] 1317 recombinant human IL-18Ra ECD with a KD of 1.5x1 O'8M or stronger.

[0565] 1318

[0566] 1319 Embodiment 101 : The VHH antibody domain or fragment thereof according to any one of

[0567] 1320 embodiments 1 to 96, wherein said VHH antibody domain or fragment thereof binds to

[0568] 1321 recombinant human IL-18Ra ECD with a KD of 1x1 O'8M or stronger.

[0569] 1322

[0570] 1323 Embodiment 102: The VHH antibody domain or fragment thereof according to any one of

[0571] 1324 embodiments 1 to 96, wherein said VHH antibody domain or fragment thereof binds to

[0572] 1325 recombinant human IL-18Ra ECD with a KD of 5x1 O'9M or stronger.

[0573] 1326

[0574] 1327 Embodiment 103: The VHH antibody domain or fragment thereof according to any one of

[0575] 1328 embodiments 1 to 96, wherein said VHH antibody domain or fragment thereof binds to

[0576] 1329 recombinant human IL-18Ra ECD with a KD of 1x1 O'9M or stronger.

[0577] 1330

[0578] 1331 Embodiment 104: The VHH antibody domain or fragment thereof according to any one of

[0579] 1332 embodiments 1 to 103, wherein said fragment of said VHH antibody domain binds to

[0580] 1333 recombinant human IL-18Ra ECD with at least 90% of the affinity (as determined by KD value)

[0581] 1334 with which said VHH antibody domain binds to recombinant human IL-18Ra ECD.

[0582] 1335

[0583] 1336 Embodiment 105: The VHH antibody domain or fragment thereof according to any one of

[0584] 1337 embodiments 1 to 104, wherein said fragment of said VHH antibody domain binds to IL-18Ra

[0585] 1338 ECD with an affinity (as determined from the KD value) that is at least equal to the affinity

[0586] 1339 with which VHH2 (SEQ ID NO: 2) binds to IL-18Ra ECD or stronger.

[0587] 1340

[0588] 1341 Such binding can be determined by in vitro binding experiments as described in Example 1

[0589] 1342 below (by biolayer interferometry).

[0590] 1343 1344 Embodiment 106: The VHH antibody domain or fragment thereof according to any one of

[0591] 1345 embodiments 1 to 23 or 62 to 105, wherein in (b) and (c) the VHH antibody domain or fragment

[0592] 1346 thereof binds to recombinant human IL-18Ra ECD with an affinity (KD value) that is by not

[0593] 1347 more than a factor of 10 weaker than the binding of the corresponding VHH antibody domain

[0594] 1348 without modification.

[0595] 1349

[0596] 1350 Embodiment 107: The VHH antibody domain or fragment thereof according to any one of

[0597] 1351 embodiments 1 to 23 or 62 to 105, wherein in (b) and (c) the VHH antibody domain or fragment

[0598] 1352 thereof binds to recombinant human IL-18Ra ECD with an affinity (KD value) that is by not

[0599] 1353 more than a factor of 5 weaker than the binding of the corresponding VHH antibody domain

[0600] 1354 without modification.

[0601] 1355

[0602] 1356 Embodiment 108: The VHH antibody domain or fragment thereof according to any one of

[0603] 1357 embodiments 1 to 23 or 62 to 105, wherein in (b) and (c) the VHH antibody domain or fragment

[0604] 1358 thereof binds to recombinant human IL-18Ra ECD with an affinity (KD value) that is by not

[0605] 1359 more than a factor of 2 weaker than the binding of the corresponding VHH antibody domain

[0606] 1360 without modification.

[0607] 1361

[0608] 1362 Embodiment 109: The VHH antibody domain or fragment thereof according to any one of

[0609] 1363 embodiments 1 to 23 or 62 to 105, wherein in (b) and (c) the VHH antibody domain or fragment

[0610] 1364 thereof binds to recombinant human IL-18Ra ECD with an affinity (KD value) that is by not

[0611] 1365 more than a factor of 1.5 weaker than the binding of the corresponding VHH antibody domain

[0612] 1366 without modification.

[0613] 1367

[0614] 1368 Embodiment 110: The VHH antibody domain or fragment thereof according to any one of

[0615] 1369 embodiments 1 to 23 or 62 to 109, wherein in (b) and (c) the VHH antibody domain or fragment

[0616] 1370 thereof binds to recombinant human IL-18Ra ECD with an affinity (KD value) that is by not

[0617] 1371 more than a factor of 10 stronger than the binding of the corresponding VHH antibody domain

[0618] 1372 without modification.

[0619] 1373

[0620] 1374 Embodiment 111 : The VHH antibody domain or fragment thereof according to any one of

[0621] 1375 embodiments 1 to 23 or 62 to 109, wherein in (b) and (c) the VHH antibody domain or fragment

[0622] 1376 thereof binds to recombinant human IL-18Ra ECD with an affinity (KD value) that is by not 1377 more than a factor of 5 stronger than the binding of the corresponding VHH antibody domain

[0623] 1378 without modification.

[0624] 1379

[0625] 1380 Embodiment 112: The VHH antibody domain or fragment thereof according to any one of

[0626] 1381 embodiments 1 to 23 or 62 to 109, wherein in (b) and (c) the VHH antibody domain or fragment

[0627] 1382 thereof binds to recombinant human IL-18Ra ECD with an affinity (KD value) that is by not

[0628] 1383 more than a factor of 2 stronger than the binding of the corresponding VHH antibody domain

[0629] 1384 without modification.

[0630] 1385

[0631] 1386 Embodiment 113: The VHH antibody domain or fragment thereof according to any one of

[0632] 1387 embodiments 1 to 23 or 62 to 109, wherein in (b) and (c) the VHH antibody domain or fragment

[0633] 1388 thereof binds to recombinant human IL-18Ra ECD with an affinity (KD value) that is by not

[0634] 1389 more than a factor of 1.5 stronger than the binding of the corresponding VHH antibody domain

[0635] 1390 without modification.

[0636] 1391

[0637] 1392 Embodiment 114: The VHH antibody domain or fragment thereof according to any one of

[0638] 1393 embodiments 24 to 113, wherein in (B) and (C) the VHH antibody domain binds to recombinant

[0639] 1394 human IL-18Ra ECD with an affinity (KD value) that is by not more than a factor of 10 weaker

[0640] 1395 than the binding of the corresponding VHH antibody domain without modification.

[0641] 1396

[0642] 1397 Embodiment 115: The VHH antibody domain or fragment thereof according to any one of

[0643] 1398 embodiments 24 to 113, wherein in (B) and (C) the VHH antibody domain binds to recombinant

[0644] 1399 human IL-18Ra ECD with an affinity (KD value) that is by not more than a factor of 5 weaker

[0645] 1400 than the binding of the corresponding VHH antibody domain without modification.

[0646] 1401

[0647] 1402 Embodiment 116: The VHH antibody domain or fragment thereof according to any one of

[0648] 1403 embodiments 24 to 113, wherein in (B) and (C) the VHH antibody domain binds to recombinant

[0649] 1404 human IL-18Ra ECD with an affinity (KD value) that is by not more than a factor of 2 weaker

[0650] 1405 than the binding of the corresponding VHH antibody domain without modification.

[0651] 1406

[0652] 1407 Embodiment 117: The VHH antibody domain or fragment thereof according to any one of

[0653] 1408 embodiments 24 to 113, wherein in (B) and (C) the VHH antibody domain binds to recombinant

[0654] 1409 human IL-18Ra ECD with an affinity (KD value) that is by not more than a factor of 1.5 weaker

[0655] 1410 than the binding of the corresponding VHH antibody domain without modification. 1411

[0656] 1412 Embodiment 118: The VHH antibody domain or fragment thereof according to any one of

[0657] 1413 embodiments 24 to 117, wherein in (B) and (C) the VHH antibody domain binds to recombinant

[0658] 1414 human IL- 18Ra ECD with an affinity (KD value) that i s by not more than a factor of 10 stronger

[0659] 1415 than the binding of the corresponding VHH antibody domain without modification.

[0660] 1416

[0661] 1417 Embodiment 119: The VHH antibody domain or fragment thereof according to any one of

[0662] 1418 embodiments 24 to 117, wherein in (B) and (C) the VHH antibody domain binds to recombinant

[0663] 1419 human IL-18Ra ECD with an affinity (KD value) that is by not more than a factor of 5 stronger

[0664] 1420 than the binding of the corresponding VHH antibody domain without modification.

[0665] 1421

[0666] 1422 Embodiment 120: The VHH antibody domain or fragment thereof according to any one of

[0667] 1423 embodiments 24 to 117, wherein in (B) and (C) the VHH antibody domain binds to recombinant

[0668] 1424 human IL-18Ra ECD with an affinity (KD value) that is by not more than a factor of 2 stronger

[0669] 1425 than the binding of the corresponding VHH antibody domain without modification.

[0670] 1426

[0671] 1427 Embodiment 121 : The VHH antibody domain or fragment thereof according to any one of

[0672] 1428 embodiments 24 to 117, wherein in (B) and (C) the VHH antibody domain binds to recombinant

[0673] 1429 human IL-18Ra ECD with an affinity (KD value) that is by not more than a factor of 1.5

[0674] 1430 stronger than the binding of the corresponding VHH antibody domain without modification.

[0675] 1431

[0676] 1432 Embodiment 122: The VHH antibody domain or fragment thereof according to any one of

[0677] 1433 embodiments 24 to 121, wherein in (D) the affinity (KD value) of the binding of the VHH

[0678] 1434 antibody domain to recombinant human IL-18RaECD is by not more than a factor of 10 weaker

[0679] 1435 than the affinity (KD value) of the binding to recombinant human IL-18Ra ECD of a VHH

[0680] 1436 antibody domain consisting of the sequence from the Table of VHH Sequences that has the

[0681] 1437 highest degree of sequence identity with the sequence of said VHH antibody domain of (D).

[0682] 1438 The degree of sequence identity can be determined by sequence alignment.

[0683] 1439

[0684] 1440 Embodiment 123: The VHH antibody domain or fragment thereof according to any one of

[0685] 1441 embodiments 24 to 121, wherein in (D) the affinity (KD value) of the binding of the VHH

[0686] 1442 antibody domain to recombinant human IL-18Ra ECD is by not more than a factor of 5 weaker

[0687] 1443 than the affinity (KD value) of the binding to recombinant human IL-18Ra ECD of a VHH

[0688] 1444 antibody domain consisting of the sequence from the Table of VHH Sequences that has the 1445 highest degree of sequence identity with the sequence of said VHH antibody domain of (D).

[0689] 1446 The degree of sequence identity can be determined by sequence alignment.

[0690] 1447

[0691] 1448 Embodiment 124: The VHH antibody domain or fragment thereof according to any one of

[0692] 1449 embodiments 24 to 121, wherein in (D) the affinity (KD value) of the binding of the VHH

[0693] 1450 antibody domain to recombinant human IL-18Ra ECD is by not more than a factor of 2 weaker

[0694] 1451 than the affinity (KD value) of the binding to recombinant human IL-18Ra ECD of a VHH

[0695] 1452 antibody domain consisting of the sequence from the Table of VHH Sequences that has the

[0696] 1453 highest degree of sequence identity with the sequence of said VHH antibody domain of (D).

[0697] 1454

[0698] 1455 Embodiment 125: The VHH antibody domain or fragment thereof according to any one of

[0699] 1456 embodiments 24 to 121, wherein in (D) the affinity (KD value) of the binding of the VHH

[0700] 1457 antibody domain to recombinant human IL-18Ra ECD is by not more than a factor of 1.5

[0701] 1458 weaker than the affinity (KD value) of the binding to recombinant human IL-18Ra ECD of a

[0702] 1459 VHH antibody domain consisting of the sequence from the Table of VHH Sequences that has

[0703] 1460 the highest degree of sequence identity with the sequence of said VHH antibody domain of (D).

[0704] 1461

[0705] 1462 Embodiment 126: The VHH antibody domain or fragment thereof according to any one of

[0706] 1463 embodiments 24 to 125, wherein in (D) the affinity (KD value) of the binding of the VHH

[0707] 1464 antibody domain to recombinant human IL-18Ra ECD is by not more than a factor of 10

[0708] 1465 stronger than the affinity (KD value) of the binding to recombinant human IL-18Ra ECD of a

[0709] 1466 VHH antibody domain consisting of the sequence from the Table of VHH Sequences that has

[0710] 1467 the highest degree of sequence identity with the sequence of said VHH antibody domain of (D).

[0711] 1468

[0712] 1469 Embodiment 127: The VHH antibody domain or fragment thereof according to any one of

[0713] 1470 embodiments 24 to 125, wherein in (D) the affinity (KD value) of the binding of the VHH

[0714] 1471 antibody domain to recombinant human IL-18Ra ECD is by not more than a factor of 5 stronger

[0715] 1472 than the affinity (KD value) of the binding to recombinant human IL-18Ra ECD of a VHH

[0716] 1473 antibody domain consisting of the sequence from the Table of VHH Sequences that has the

[0717] 1474 highest degree of sequence identity with the sequence of said VHH antibody domain of (D).

[0718] 1475

[0719] 1476 Embodiment 128: The VHH antibody domain or fragment thereof according to any one of

[0720] 1477 embodiments 24 to 125, wherein in (D) the affinity (KD value) of the binding of the VHH

[0721] 1478 antibody domain to recombinant human IL-18Ra ECD is by not more than a factor of 2 stronger 1479 than the affinity (KD value) of the binding to recombinant human IL-18Ra ECD of a VHH

[0722] 1480 antibody domain consisting of the sequence from the Table of VHH Sequences that has the

[0723] 1481 highest degree of sequence identity with the sequence of said VHH antibody domain of (D).

[0724] 1482

[0725] 1483 Embodiment 129: The VHH antibody domain or fragment thereof according to any one of

[0726] 1484 embodiments 24 to 125, wherein in (D) the affinity (KD value) of the binding of the VHH

[0727] 1485 antibody domain to recombinant human IL-18Ra ECD is by not more than a factor of 1.5

[0728] 1486 stronger than the affinity (KD value) of the binding to recombinant human IL-18Ra ECD of a

[0729] 1487 VHH antibody domain consisting of the sequence from the Table of VHH Sequences that has

[0730] 1488 the highest degree of sequence identity with the sequence of said VHH antibody domain of (D).

[0731] 1489

[0732] 1490 Embodiment 130: The VHH antibody domain or fragment thereof according to any one of

[0733] 1491 embodiments 91 to 129, wherein said KD value is determined in in vitro binding experiments

[0734] 1492 through biolayer interferometry.

[0735] 1493

[0736] 1494 Embodiment 131 : The VHH antibody domain or fragment thereof according to any one of

[0737] 1495 embodiments 91 to 130, wherein said KD value is measured by kinetic measurements by

[0738] 1496 biolayer interferometry at 25°C and 1000 rpm in KB Buffer (PBS + 0.1 % Tween-20 +

[0739] 1497 1% BSA).

[0740] 1498

[0741] 1499 Embodiment 132: The VHH antibody domain or fragment thereof according to any one of

[0742] 1500 embodiments 24 to 131, wherein said VHH antibody domain of (A) comprises one VHH

[0743] 1501 sequence selected from the group consisting of VHH1, VHH2, VHH3, VHH4, VHH5, VHH6,

[0744] 1502 VHH8, VHH9 and VHH10 shown in the Table of VHH Sequences.

[0745] 1503

[0746] 1504 Embodiment 133: The VHH antibody domain or fragment thereof according to any one of

[0747] 1505 embodiments 24 to 131, wherein said VHH antibody domain of (A) comprises one VHH

[0748] 1506 sequence selected from the group consisting of VHH1, VHH2, VHH5, VHH6, VHH8, VHH9

[0749] 1507 and VHH10 shown in the Table of VHH Sequences.

[0750] 1508

[0751] 1509 Embodiment 134: The VHH antibody domain or fragment thereof according to any one of

[0752] 1510 embodiments 24 to 131, wherein said VHH antibody domain of (A) comprises one VHH

[0753] 1511 sequence selected from the group consisting of VHH2, VHH6, VHH8 and VHH10 shown in

[0754] 1512 the Table of VHH Sequences. 1513

[0755] 1514 Embodiment 135: The VHH antibody domain or fragment thereof according to any one of

[0756] 1515 embodiments 24 to 131, wherein said VHH antibody domain of (A) comprises one VHH

[0757] 1516 sequence selected from the group consisting of VHH6 and VHHl 1 shown in the Table of VHH

[0758] 1517 Sequences.

[0759] 1518

[0760] 1519 Embodiment 136: The VHH antibody domain or fragment thereof according to any one of

[0761] 1520 embodiments 24 to 131, wherein said VHH antibody domain of (A) comprises one VHH

[0762] 1521 sequence selected from the group consisting of VHHl, VHH2, VHH5, VHH6, VHH8 and

[0763] 1522 VHH10 shown in the Table of VHH Sequences.

[0764] 1523

[0765] 1524 Embodiment 137: The VHH antibody domain or fragment thereof according to any one of

[0766] 1525 embodiments 24 to 131, wherein said VHH antibody domain of (A) comprises one VHH

[0767] 1526 sequence selected from the group consisting of VHH2, VHH6, VHH8 and VHH10 shown in

[0768] 1527 the Table of VHH Sequences.

[0769] 1528

[0770] 1529 Embodiment 138: The VHH antibody domain or fragment thereof according to any one of

[0771] 1530 embodiments 24 to 131, wherein said VHH antibody domain of (A) comprises one VHH

[0772] 1531 sequence selected from the group consisting of VHH2 and VHH8 shown in the Table of VHH

[0773] 1532 Sequences.

[0774] 1533

[0775] 1534 Embodiment 139: The VHH antibody domain or fragment thereof according to any one of

[0776] 1535 embodiments 24 to 131, wherein said VHH antibody domain of (A) comprises the VHH

[0777] 1536 sequence VHHl shown in the Table of VHH Sequences.

[0778] 1537

[0779] 1538 Embodiment 140: The VHH antibody domain or fragment thereof according to any one of

[0780] 1539 embodiments 24 to 131, wherein said VHH antibody domain of (A) comprises the VHH

[0781] 1540 sequence VHH2 shown in the Table of VHH Sequences.

[0782] 1541

[0783] 1542 Embodiment 141 : The VHH antibody domain or fragment thereof according to any one of

[0784] 1543 embodiments 24 to 131, wherein said VHH antibody domain of (A) comprises the VHH

[0785] 1544 sequence VHH3 shown in the Table of VHH Sequences.

[0786] 1545 1546 Embodiment 142: The VHH antibody domain or fragment thereof according to any one of

[0787] 1547 embodiments 24 to 131, wherein said VHH antibody domain of (A) comprises the VHH

[0788] 1548 sequence VHH4 shown in the Table of VHH Sequences.

[0789] 1549

[0790] 1550 Embodiment 143: The VHH antibody domain or fragment thereof according to any one of

[0791] 1551 embodiments 24 to 131, wherein said VHH antibody domain of (A) comprises the VHH

[0792] 1552 sequence VHH5 shown in the Table of VHH Sequences.

[0793] 1553

[0794] 1554 Embodiment 144: The VHH antibody domain or fragment thereof according to any one of

[0795] 1555 embodiments 24 to 131, wherein said VHH antibody domain of (A) comprises the VHH

[0796] 1556 sequence VHH6 shown in the Table of VHH Sequences.

[0797] 1557

[0798] 1558 Embodiment 145: The VHH antibody domain or fragment thereof according to any one of

[0799] 1559 embodiments 24 to 131, wherein said VHH antibody domain of (A) comprises the VHH

[0800] 1560 sequence VHH8 shown in the Table of VHH Sequences.

[0801] 1561

[0802] 1562 Embodiment 146: The VHH antibody domain or fragment thereof according to any one of

[0803] 1563 embodiments 24 to 131, wherein said VHH antibody domain of (A) comprises the VHH

[0804] 1564 sequence VHH9 shown in the Table of VHH Sequences.

[0805] 1565

[0806] 1566 Embodiment 147: The VHH antibody domain or fragment thereof according to any one of

[0807] 1567 embodiments 24 to 131, wherein said VHH antibody domain of (A) comprises the VHH

[0808] 1568 sequence VHH10 shown in the Table of VHH Sequences.

[0809] 1569

[0810] 1570 Embodiment 148: The VHH antibody domain or fragment thereof according to any one of

[0811] 1571 embodiments 24 to 131, wherein said VHH antibody domain of (A) comprises the VHH

[0812] 1572 sequence VHH11 shown in the Table of VHH Sequences.

[0813] 1573

[0814] 1574 Embodiment 149: The VHH antibody domain or fragment thereof according to any one of

[0815] 1575 embodiments 1 to 23 or 62 to 148, wherein in (a) said VHH antibody domain or fragment

[0816] 1576 thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of one

[0817] 1577 VHH selected from the group consisting of VHH1, VHH2, VHH3, VHH4, VHH5, VHH6,

[0818] 1578 VHH8, VHH9 and VHH10 as shown in the Table of CDRs.

[0819] 1579 1580 Embodiment 150: The VHH antibody domain or fragment thereof according to any one of

[0820] 1581 embodiments 1 to 23 or 62 to 148, wherein in (a) said VHH antibody domain or fragment

[0821] 1582 thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of one

[0822] 1583 VHH selected from the group consisting of VHH1, VHH2, VHH5, VHH6, VHH8, VHH9 and

[0823] 1584 VHH10 as shown in the Table of CDRs.

[0824] 1585

[0825] 1586 Embodiment 151 : The VHH antibody domain or fragment thereof according to any one of

[0826] 1587 embodiments 1 to 23 or 62 to 148, wherein in (a) said VHH antibody domain or fragment

[0827] 1588 thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of

[0828] 1589 VHH2, VHH6, VHH8 and VHH10 as shown in the Table of CDRs.

[0829] 1590

[0830] 1591 Embodiment 152: The VHH antibody domain or fragment thereof according to any one of

[0831] 1592 embodiments 1 to 23 or 62 to 148, wherein in (a) said VHH antibody domain or fragment

[0832] 1593 thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of one

[0833] 1594 VHH selected from the group consisting of VHH6 and VHH11.

[0834] 1595

[0835] 1596 Embodiment 153: The VHH antibody domain or fragment thereof according to any one of

[0836] 1597 embodiments 1 to 23 or 62 to 148, wherein in (a) said VHH antibody domain or fragment

[0837] 1598 thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of one

[0838] 1599 VHH selected from the group consisting of VHH1, VHH2, VHH5, VHH6, VHH8 and VHH10

[0839] 1600 as shown in the Table of CDRs.

[0840] 1601

[0841] 1602 Embodiment 154: The VHH antibody domain or fragment thereof according to any one of

[0842] 1603 embodiments 1 to 23 or 62 to 148, wherein in (a) said VHH antibody domain or fragment

[0843] 1604 thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of one

[0844] 1605 VHH selected from the group consisting of VHH2, VHH6, VHH8, VHH10 as shown in the

[0845] 1606 Table of CDRs.

[0846] 1607

[0847] 1608 Embodiment 155: The VHH antibody domain or fragment thereof according to any one of

[0848] 1609 embodiments 1 to 23 or 62 to 148, wherein in (a) said VHH antibody domain or fragment

[0849] 1610 thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of one

[0850] 1611 VHH selected from the group consisting of VHH2 or VHH8 as shown in the Table of CDRs.

[0851] 1612 1613 Embodiment 156: The VHH antibody domain or fragment thereof according to any one of

[0852] 1614 embodiments 1 to 23 or 62 to 148, wherein in (a) said VHH antibody domain or fragment

[0853] 1615 thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of VHH1

[0854] 1616 as shown in the Table of CDRs.

[0855] 1617

[0856] 1618 Embodiment 157: The VHH antibody domain or fragment thereof according to any one of

[0857] 1619 embodiments 1 to 23 or 62 to 148, wherein in (a) said VHH antibody domain or fragment

[0858] 1620 thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of VHH2

[0859] 1621 as shown in the Table of CDRs.

[0860] 1622

[0861] 1623 Embodiment 158: The VHH antibody domain or fragment thereof according to any one of

[0862] 1624 embodiments 1 to 23 or 62 to 148, wherein in (a) said VHH antibody domain or fragment

[0863] 1625 thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of VHH3

[0864] 1626 as shown in the Table of CDRs.

[0865] 1627

[0866] 1628 Embodiment 159: The VHH antibody domain or fragment thereof according to any one of

[0867] 1629 embodiments 1 to 23 or 62 to 148, wherein in (a) said VHH antibody domain or fragment

[0868] 1630 thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of VHH4

[0869] 1631 as shown in the Table of CDRs.

[0870] 1632

[0871] 1633 Embodiment 160: The VHH antibody domain or fragment thereof according to any one of

[0872] 1634 embodiments 1 to 23 or 62 to 148, wherein in (a) said VHH antibody domain or fragment

[0873] 1635 thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of VHH5

[0874] 1636 as shown in the Table of CDRs.

[0875] 1637

[0876] 1638 Embodiment 161 : The VHH antibody domain or fragment thereof according to any one of

[0877] 1639 embodiments 1 to 23 or 62 to 148, wherein in (a) said VHH antibody domain or fragment

[0878] 1640 thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of VHH6

[0879] 1641 as shown in the Table of CDRs.

[0880] 1642

[0881] 1643 Embodiment 162: The VHH antibody domain or fragment thereof according to any one of

[0882] 1644 embodiments 1 to 23 or 62 to 148, wherein in (a) said VHH antibody domain or fragment

[0883] 1645 thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of VHH8

[0884] 1646 as shown in the Table of CDRs. 1647

[0885] 1648 Embodiment 163: The VHH antibody domain or fragment thereof according to any one of

[0886] 1649 embodiments 1 to 23 or 62 to 148, wherein in (a) said VHH antibody domain or fragment

[0887] 1650 thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of VHH9

[0888] 1651 as shown in the Table of CDRs.

[0889] 1652

[0890] 1653 Embodiment 164: The VHH antibody domain or fragment thereof according to any one of

[0891] 1654 embodiments 1 to 23 or 62 to 148, wherein in (a) said VHH antibody domain or fragment

[0892] 1655 thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of

[0893] 1656 VHH10 as shown in the Table of CDRs.

[0894] 1657

[0895] 1658 Embodiment 165: The VHH antibody domain or fragment thereof according to any one of

[0896] 1659 embodiments 1 to 23 or 62 to 148, wherein in (a) said VHH antibody domain or fragment

[0897] 1660 thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of

[0898] 1661 VHH11 as shown in the Table of CDRs.

[0899] 1662

[0900] 1663 Embodiment 166: The VHH antibody domain or fragment thereof according to any one of

[0901] 1664 embodiments 1 to 165, wherein said VHH antibody domain or fragment thereof competes with

[0902] 1665 VHH2 for binding to recombinant human IL-18Ra ECD.

[0903] 1666

[0904] 1667 Embodiment 167: The VHH antibody domain or fragment thereof according to any one of

[0905] 1668 embodiments 1 to 165, wherein said VHH antibody domain or fragment thereof partially

[0906] 1669 competes with VHH2 for binding to recombinant human IL-18Ra ECD.

[0907] 1670

[0908] 1671 Embodiment 168: The VHH antibody domain or fragment thereof according to any one of

[0909] 1672 embodiments 1 to 167, wherein said VHH antibody domain or fragment thereof does not

[0910] 1673 compete with VHH15 for binding to recombinant human IL-18RP ECD.

[0911] 1674

[0912] 1675 Embodiment 169: The VHH antibody domain or fragment thereof according to any one of

[0913] 1676 embodiments 1 to 168, wherein said humanization is by germlining.

[0914] 1677

[0915] 1678 Embodiment 170: The VHH antibody domain or fragment thereof according to embodiment

[0916] 1679 169, wherein said germlining involves 1680 - identifying the human VH germline sequence that is closest to the sequence to be

[0917] 1681 humanized with respect to its sequence similarity; and

[0918] 1682 - replacing amino acid(s) in the sequence to be humanized by the amino acid(s) at the

[0919] 1683 corresponding sequence position(s) in the closest human VH germline sequence.

[0920] 1684

[0921] 1685 Sequence similarity and corresponding sequence positions can be determined by sequence

[0922] 1686 alignment as described above.

[0923] 1687

[0924] 1688 Embodiment 171 : The VHH antibody domain or fragment thereof according to embodiment

[0925] 1689 169, wherein said germlining consists of

[0926] 1690 - identifying the human VH germline sequence that is closest to the sequence to be

[0927] 1691 humanized with respect to its sequence similarity; and

[0928] 1692 - replacing amino acid(s) in the sequence to be humanized by the amino acid(s) at the

[0929] 1693 corresponding sequence position(s) in the closest human VH germline sequence.

[0930] 1694

[0931] 1695 Embodiment 172: The VHH antibody domain or fragment thereof according to any one of

[0932] 1696 embodiments 1 to 171, wherein said VHH antibody domain (resp. fragment thereof) with

[0933] 1697 humanization binds to human IL-18Ra ECD with an affinity (KD value) that is not weaker by

[0934] 1698 a factor of more than 10 compared to the binding of a corresponding VHH antibody domain

[0935] 1699 (resp. fragment thereof) without humanization to human IL-18Ra ECD.

[0936] 1700

[0937] 1701 As a skilled person understands, the term "corresponding VHH antibody domain without

[0938] 1702 humanization" refers to a VHH antibody domain that differs only by the amino acid changes

[0939] 1703 introduced in order to carry out said humanization, but that is otherwise identical.

[0940] 1704

[0941] 1705 Embodiment 173: The VHH antibody domain or fragment thereof according to any one of

[0942] 1706 embodiments 1 to 171, wherein said VHH antibody domain (resp. fragment thereof) with

[0943] 1707 humanization binds to human IL-18Ra ECD with an affinity (KD value) that is not weaker by

[0944] 1708 a factor of more than 5 compared to the binding of a corresponding VHH antibody domain

[0945] 1709 (resp. fragment thereof) without humanization to human IL-18Ra ECD.

[0946] 1710

[0947] 1711 Embodiment 174: The VHH antibody domain or fragment thereof according to any one of

[0948] 1712 embodiments 1 to 171, wherein said VHH antibody domain (resp. fragment thereof) with

[0949] 1713 humanization binds to human IL-18Ra ECD with an affinity (KD value) that is not weaker by 1714 a factor of more than 2 compared to the binding of a corresponding VHH antibody domain

[0950] 1715 (resp. fragment thereof) without humanization to human IL-18Ra ECD.

[0951] 1716

[0952] 1717 Embodiment 175: The VHH antibody domain or fragment thereof according to any one of

[0953] 1718 embodiments 1 to 171, wherein said VHH antibody domain (resp. fragment thereof) with

[0954] 1719 humanization binds to human IL-18Ra ECD with an affinity (KD value) that is not weaker than

[0955] 1720 the binding of a corresponding VHH antibody domain (resp. fragment thereof) without

[0956] 1721 humanization to human IL-18Ra ECD.

[0957] 1722

[0958] 1723 Embodiment 176: The VHH antibody domain or fragment thereof according to any one of

[0959] 1724 embodiments 172 to 175, wherein said affinity is determined by KD measurement.

[0960] 1725

[0961] 1726 Embodiment 177: The VHH antibody domain or fragment thereof according to any one of

[0962] 1727 embodiments 24 to 176, wherein said VHH antibody domain comprises a VHH antibody

[0963] 1728 domain according to (A), (B) or (C) / said fragment of a VHH antibody domain comprises a

[0964] 1729 fragment of a VHH antibody domain according to (A), (B) or (C).

[0965] 1730

[0966] 1731 Embodiment 178: The VHH antibody domain or fragment thereof according to any one of

[0967] 1732 embodiments 24 to 176, wherein said VHH antibody domain comprises a VHH antibody

[0968] 1733 domain according to (A), (B) or (D) / said fragment of a VHH antibody domain comprises a

[0969] 1734 fragment of a VHH antibody domain according to (A), (B) or (D).

[0970] 1735

[0971] 1736 Embodiment 179: The VHH antibody domain or fragment thereof according to any one of

[0972] 1737 embodiments 24 to 176, wherein said VHH antibody domain comprises a VHH antibody

[0973] 1738 domain according to (A), (C) or (D) / said fragment of a VHH antibody domain comprises a

[0974] 1739 fragment of a VHH antibody domain according to (A), (C) or (D).

[0975] 1740

[0976] 1741 Embodiment 180: The VHH antibody domain or fragment thereof according to any one of

[0977] 1742 embodiments 24 to 176, wherein said VHH antibody domain comprises a VHH antibody

[0978] 1743 domain according to (A) or (B) / said fragment of a VHH antibody domain comprises a fragment

[0979] 1744 of a VHH antibody domain according to (A) or (B).

[0980] 1745

[0981] 1746 Embodiment 181 : The VHH antibody domain or fragment thereof according to any one of

[0982] 1747 embodiments 24 to 176, wherein said VHH antibody domain comprises a VHH antibody 1748 domain according to (A) or (C) / said fragment of a VHH antibody domain comprises a fragment

[0983] 1749 of a VHH antibody domain according to (A) or (C).

[0984] 1750

[0985] 1751 Embodiment 182: The VHH antibody domain or fragment thereof according to any one of

[0986] 1752 embodiments 24 to 176, wherein said VHH antibody domain comprises a VHH antibody

[0987] 1753 domain according to (A) or (D) / said fragment of a VHH antibody domain comprises a fragment

[0988] 1754 of a VHH antibody domain according to (A) or (D).

[0989] 1755

[0990] 1756 Embodiment 183: The VHH antibody domain or fragment thereof according to any one of

[0991] 1757 embodiments 24 to 176, wherein said VHH antibody domain comprises a VHH antibody

[0992] 1758 domain according to (A) / said fragment of a VHH antibody domain comprises a fragment of a

[0993] 1759 VHH antibody domain according to (A).

[0994] 1760

[0995] 1761 Embodiment 184: The VHH antibody domain or fragment thereof according to any one of

[0996] 1762 embodiments 24 to 176, wherein said VHH antibody domain comprises a VHH antibody

[0997] 1763 domain according to (B) / said fragment of a VHH antibody domain comprises a fragment of a

[0998] 1764 VHH antibody domain according to (B).

[0999] 1765

[1000] 1766 Embodiment 185: The VHH antibody domain or fragment thereof according to any one of

[1001] 1767 embodiments 1 to 23 or 62 to 184, wherein said VHH antibody domain or fragment thereof

[1002] 1768 comprises complementarity determining regions according to (a) or (b).

[1003] 1769

[1004] 1770 Embodiment 186: The VHH antibody domain or fragment thereof according to any one of

[1005] 1771 embodiments 1 to 23 or 62 to 184, wherein said VHH antibody domain or fragment thereof

[1006] 1772 comprises complementarity determining regions according to (a) or (c).

[1007] 1773

[1008] 1774 Embodiment 187: The VHH antibody domain or fragment thereof according to any one of

[1009] 1775 embodiments 1 to 23 or 62 to 184, wherein said VHH antibody domain or fragment thereof

[1010] 1776 comprises complementarity determining regions according to (a).

[1011] 1777

[1012] 1778 Embodiment 188: The VHH antibody domain or fragment thereof according to any one of

[1013] 1779 embodiments 1 to 23 or 62 to 184, wherein said VHH antibody domain or fragment thereof

[1014] 1780 comprises complementarity determining regions according to (b).

[1015] 1781 1782 Fourth aspect of the present disclosure (also referred to as "Embodiment 189"): According to a

[1016] 1783 fourth aspect, the present disclosure relates to a VHH antibody domain or a fragment thereof,

[1017] 1784 wherein

[1018] 1785 (d) said VHH antibody domain or fragment thereof comprises the complementarity

[1019] 1786 determining regions CDR1, CDR2 and CDR3 of one VHH selected from the group

[1020] 1787 consisting of VHH12, VHH13, VHH14, VHH15, VHH16, VHH17, VHH18,

[1021] 1788 VHH20, VHH21 and VHH22 as shown in the Table of CDRs;

[1022] 1789 (e) said VHH antibody domain or fragment thereof comprises the complementarity

[1023] 1790 determining regions CDR1, CDR2 and CDR3 as defined in (d) with modification,

[1024] 1791 wherein the modification is that the sequence of at least one of CDR1, CDR2 and

[1025] 1792 CDR3 is humanized; or

[1026] 1793 (f) said VHH antibody domain or fragment thereof comprises the complementarity

[1027] 1794 determining regions CDR1, CDR2 and CDR3 as defined in (d) with modification,

[1028] 1795 wherein the modification is

[1029] 1796 - the replacement, addition or deletion of up to three amino acids in CDR1,

[1030] 1797 - the replacement, addition or deletion of up to three amino acids in CDR2

[1031] 1798 and / or

[1032] 1799 - the replacement, addition or deletion of up to three amino acids in CDR3;

[1033] 1800

[1034] 1801 Table of CDRs:

[1035] 1802

[1036] 1803

[1037] 1804 To the fourth aspect of the present disclosure and the embodiments referring thereto, the same

[1038] 1805 explanations and definitions apply accordingly as for the first to third aspects of the present

[1039] 1806 disclosure and the embodiments referring thereto.

[1040] 1807

[1041] 1808 Embodiment 190: The VHH antibody domain or fragment thereof according to embodiment

[1042] 1809 189, wherein the modification in (e) is that the sequence of CDR1 and / or CDR2, but not the

[1043] 1810 sequence of CDR3 is humanized.

[1044] 1811 1812 Embodiment 191 : The VHH antibody domain or fragment thereof according to any one of

[1045] 1813 embodiments 189 or 190, wherein the modification in (e) is that the sequence of CDR1 is

[1046] 1814 humanized, but not the sequence of CDR2 and CDR3.

[1047] 1815

[1048] 1816 Embodiment 192: The VHH antibody domain or fragment thereof according to any one of

[1049] 1817 embodiments 189 or 190, wherein the modification in (e) is that the sequence of CDR2 is

[1050] 1818 humanized, but not the sequence of CDR1 and CDR3.

[1051] 1819

[1052] 1820 Embodiment 193: The VHH antibody domain or fragment thereof according to any one of

[1053] 1821 embodiments 189 to 192, wherein the modification in (e) is that the sequence of one, but not

[1054] 1822 more than one of CDR1, CDR2 and CDR3 is humanized.

[1055] 1823

[1056] 1824 Embodiment 194: The VHH antibody domain or fragment thereof according to any one of

[1057] 1825 embodiments 189 to 193, wherein said humanization of said CDR(s) is by replacing at least one

[1058] 1826 amino acid in the sequence of said CDR by the corresponding amino acid of a human VH

[1059] 1827 domain.

[1060] 1828

[1061] 1829 Embodiment 195: The VHH antibody domain or fragment thereof according to any one of

[1062] 1830 embodiments 189 to 194, wherein said humanization of said CDR(s) is by replacing up to three

[1063] 1831 amino acids in the sequence of said CDR by the corresponding amino acid of a human VH

[1064] 1832 domain.

[1065] 1833

[1066] 1834 Embodiment 196: The VHH antibody domain or fragment thereof according to any one of

[1067] 1835 embodiments 189 to 194, wherein said humanization of said CDR(s) is by replacing up to three

[1068] 1836 amino acids in the sequence of CDR1 and / or CDR2 and up to one amino acid in the sequence

[1069] 1837 of CDR3 by the corresponding amino acid of a human VH domain.

[1070] 1838

[1071] 1839 Embodiment 197: The VHH antibody domain or fragment thereof according to any one of

[1072] 1840 embodiments 189 to 194, wherein said humanization of said CDR(s) is by replacing up to two

[1073] 1841 amino acids in the sequence of said CDR by the corresponding amino acid of a human VH

[1074] 1842 domain.

[1075] 1843

[1076] 1844 Embodiment 198: The VHH antibody domain or fragment thereof according to any one of

[1077] 1845 embodiments 189 to 194, wherein said humanization of said CDR(s) is by replacing up to two 1846 amino acids in the sequence of CDR1 and / or CDR2 and up to one amino acid in the sequence

[1078] 1847 of CDR3 by the corresponding amino acid of a human VH domain.

[1079] 1848

[1080] 1849 Embodiment 199: The VHH antibody domain or fragment thereof according to any one of

[1081] 1850 embodiments 189 to 198, wherein said humanization of said CDR(s) is by replacing one amino

[1082] 1851 acid in the sequence of said CDR by the corresponding amino acid of a human VH domain.

[1083] 1852

[1084] 1853 Embodiment 200: The VHH antibody domain or fragment thereof according to any one of

[1085] 1854 embodiments 189 to 199, wherein the modification in (f) is

[1086] 1855 - the replacement, addition or deletion of up to three amino acids in CDR1,

[1087] 1856 - the replacement, addition or deletion of up to three amino acids in CDR2 and / or

[1088] 1857 - the replacement, addition or deletion of up to one amino acid in CDR3.

[1089] 1858

[1090] 1859 Embodiment 201 : The VHH antibody domain or fragment thereof according to any one of

[1091] 1860 embodiments 189 to 199, wherein the modification in (f) is

[1092] 1861 - the replacement, addition or deletion of up to two amino acids in CDR1,

[1093] 1862 - the replacement, addition or deletion of up to two amino acids in CDR2 and / or

[1094] 1863 - the replacement, addition or deletion of up to two amino acids in CDR3;

[1095] 1864

[1096] 1865 Embodiment 202: The VHH antibody domain or fragment thereof according to any one of

[1097] 1866 embodiments 189 to 199, wherein the modification in (f) is

[1098] 1867 - the replacement, addition or deletion of up to two amino acids in CDR1;

[1099] 1868 - the replacement, addition or deletion of up to two amino acids in CDR2; and / or

[1100] 1869 - the replacement, addition or deletion of up to one amino acid in CDR3.

[1101] 1870

[1102] 1871 Embodiment 203: The VHH antibody domain or fragment thereof according to any one of

[1103] 1872 embodiments 189 to 199, wherein the modification in (f) is

[1104] 1873 - the replacement, addition or deletion of up to two amino acids in CDR1 and / or

[1105] 1874 - the replacement, addition or deletion of up to two amino acids in CDR2;

[1106] 1875 wherein the sequence of CDR3 is unmodified.

[1107] 1876

[1108] 1877 As a skilled person understands, the indication that the "sequence of CDR3 is unmodified"

[1109] 1878 means that the sequence is unmodified compared to the sequence provided for CDR3 for the

[1110] 1879 VHH at issue in the Table of CDRs. 1880

[1111] 1881 Embodiment 204: The VHH antibody domain or fragment thereof according to any one of

[1112] 1882 embodiments 189 to 199, wherein the modification in (f) is

[1113] 1883 - the replacement, addition or deletion of up to two amino acids in CDR1,

[1114] 1884 wherein the sequence of CDR2 and CDR3 is unmodified.

[1115] 1885

[1116] 1886 Embodiment 205: The VHH antibody domain or fragment thereof according to any one of

[1117] 1887 embodiments 189 to 199, wherein the modification in (f) is

[1118] 1888 - the replacement, addition or deletion of up to two amino acids in CDR2,

[1119] 1889 wherein the sequence of CDR1 and CDR3 is unmodified.

[1120] 1890

[1121] 1891 Embodiment 206: The VHH antibody domain or fragment thereof according to any one of

[1122] 1892 embodiments 189 to 199, wherein the modification in (f) is

[1123] 1893 - the replacement, addition or deletion of up to one amino acid in CDR1;

[1124] 1894 - the replacement, addition or deletion of up to one amino acid in CDR2; and / or

[1125] 1895 - the replacement, addition or deletion of up to one amino acid in CDR3.

[1126] 1896

[1127] 1897 Embodiment 207: The VHH antibody domain or fragment thereof according to any one of

[1128] 1898 embodiments 189 to 199, wherein the modification in (f) is

[1129] 1899 - the replacement, addition or deletion of one amino acid in CDR1 and / or

[1130] 1900 - the replacement, addition or deletion of one amino acid in CDR2;

[1131] 1901 wherein the sequence of CDR3 is unmodified.

[1132] 1902

[1133] 1903 Embodiment 208: The VHH antibody domain or fragment thereof according to any one of

[1134] 1904 embodiments 189 to 199, wherein the modification in (f) is

[1135] 1905 - the replacement, addition or deletion of one amino acid in CDR1,

[1136] 1906 wherein the sequence of CDR2 and CDR3 is unmodified.

[1137] 1907

[1138] 1908 Embodiment 209: The VHH antibody domain or fragment thereof according to any one of

[1139] 1909 embodiments 189 to 199, wherein the modification in (f) is

[1140] 1910 - the replacement, addition or deletion of one amino acid in CDR2,

[1141] 1911 wherein the sequence of CDR1 and CDR3 is unmodified.

[1142] 1912 1913 Embodiment 210: The VHH antibody domain or fragment thereof according to any one of

[1143] 1914 embodiments 189 to 209, wherein the modification in (f) comprises only the replacement, but

[1144] 1915 not the addition or deletion of amino acids.

[1145] 1916

[1146] 1917 Embodiment 211 : The VHH antibody domain or fragment thereof according to any one of

[1147] 1918 embodiments 189 to 210, wherein said replacement is a conservative amino acid replacement.

[1148] 1919

[1149] 1920 Fifth aspect of the present disclosure (also referred to as "Embodiment 212"): According to a

[1150] 1921 fifth aspect, the present disclosure relates to a VHH antibody domain or a fragment thereof,

[1151] 1922 wherein

[1152] 1923 (E) said VHH antibody domain comprises the amino acid sequence of a VHH selected

[1153] 1924 from the group consisting of VHH12, VHH13, VHH14, VHH15, VHH16, VHH17,

[1154] 1925 VHH18, VHH20, VHH21 and VHH22 as shown in the Table of VHH Sequences;

[1155] 1926 (F) said VHH antibody domain comprises a VHH sequence as defined in (E) with

[1156] 1927 modification, wherein the modification is that said sequence is humanized;

[1157] 1928 (G) said VHH antibody domain comprises a VHH sequence as defined in (E) with

[1158] 1929 modification, wherein the modification is the replacement, addition or deletion of up

[1159] 1930 to 25 amino acids; or

[1160] 1931 (H) said VHH antibody domain comprises a VHH sequence that is at least 75% identical

[1161] 1932 to a VHH sequence referred to in (E);

[1162] 1933

[1163] 1934 Table of VHH Sequences:

[1164] 1935

[1165] 1936

[1166] 1937 Sixth aspect of the present disclosure (also referred to as "Embodiment 213"): According to a

[1167] 1938 sixth aspect, the present disclosure relates to a VHH antibody domain or a fragment thereof,

[1168] 1939 wherein

[1169] 1940 (E) said VHH antibody domain consists of the amino acid sequence of a VHH selected

[1170] 1941 from the group consisting of VHH12, VHH13, VHH14, VHH15, VHH16, VHH17,

[1171] 1942 VHH18, VHH20, VHH21 and VHH22 as shown in the Table of VHH Sequences;

[1172] 1943 (F) said VHH antibody domain consists of a VHH sequence as defined in (E) with

[1173] 1944 modification, wherein the modification is that said sequence is humanized;

[1174] 1945 (G) said VHH antibody domain consists of a VHH sequence as defined in (E) with

[1175] 1946 modification, wherein the modification is the replacement, addition or deletion of up

[1176] 1947 to 25 amino acids; or 1948 (H) said VHH antibody domain consists of a VHH sequence that is at least 75% identical

[1177] 1949 to a VHH sequence referred to in (E).

[1178] 1950

[1179] 1951 To the fifth and sixth aspects of the present disclosure and the embodiments referring thereto,

[1180] 1952 the same explanations and definitions apply accordingly as for the first to fourth aspects of the

[1181] 1953 present disclosure and the embodiments referring thereto.

[1182] 1954

[1183] 1955 Embodiment 214: The VHH antibody domain or fragment thereof according to any one of

[1184] 1956 embodiments 212 or 213, wherein said fragment of said VHH antibody domain comprises at

[1185] 1957 least 75% of the amino acids of the sequence of said VHH antibody domain. As the skilled

[1186] 1958 person understands, this means that that fragment lacks up to a quarter of the total number of

[1187] 1959 amino acids of said VHH antibody domain, wherein, compared to the "complete" VHH

[1188] 1960 antibody domain sequence said amino acids are lacking either at the N-terminus or at the C-

[1189] 1961 terminus.

[1190] 1962

[1191] 1963 Embodiment 215: The VHH antibody domain or fragment thereof according to any one of

[1192] 1964 embodiments 212 or 213, wherein said fragment of said VHH antibody domain comprises at

[1193] 1965 least 80% of the amino acids of the sequence of said VHH antibody domain.

[1194] 1966

[1195] 1967 Embodiment 216: The VHH antibody domain or fragment thereof according to any one of

[1196] 1968 embodiments 212 or 213, wherein said fragment of said VHH antibody domain comprises at

[1197] 1969 least 85% of the amino acids of the sequence of said VHH antibody domain.

[1198] 1970

[1199] 1971 Embodiment 217: The VHH antibody domain or fragment thereof according to any one of

[1200] 1972 embodiments 212 or 213, wherein said fragment of said VHH antibody domain comprises at

[1201] 1973 least 90% of the amino acids of the sequence of said VHH antibody domain.

[1202] 1974

[1203] 1975 Embodiment 218: The VHH antibody domain or fragment thereof according to any one of

[1204] 1976 embodiments 212 or 213, wherein said fragment of said VHH antibody domain comprises at

[1205] 1977 least 95% of the amino acids of the sequence of said VHH antibody domain.

[1206] 1978

[1207] 1979 Embodiment 219: The VHH antibody domain or fragment thereof according to any one of

[1208] 1980 embodiments 212 or 213, wherein said fragment of said VHH antibody domain comprises at

[1209] 1981 least 98% of the amino acids of the sequence of said VHH antibody domain. 1982

[1210] 1983 Embodiment 220: The VHH antibody domain or fragment thereof according to any one of

[1211] 1984 embodiments 212 or 213, wherein said fragment of said VHH antibody domain comprises at

[1212] 1985 least 99% of the amino acids of the sequence of said VHH antibody domain.

[1213] 1986

[1214] 1987 Embodiment 221 : The VHH antibody domain or fragment thereof according to any one of

[1215] 1988 embodiments 212 to 220, wherein said fragment of said VHH antibody domain comprises the

[1216] 1989 complementarity determining regions CDR1, CDR2 and CDR3.

[1217] 1990

[1218] 1991 Embodiment 222: The VHH antibody domain or fragment thereof according to any one of

[1219] 1992 embodiments 212 to 221, wherein said fragment of said VHH antibody domain comprises at

[1220] 1993 least the sequence from the N-terminus of CDR1 to the C-terminus of CDR3 of said VHH

[1221] 1994 antibody domain.

[1222] 1995

[1223] 1996 Embodiment 223: The VHH antibody domain or fragment thereof according to any one of

[1224] 1997 embodiments 212 to 222, wherein in (E) said fragment of said VHH antibody domain comprises

[1225] 1998 all the complementarity determining regions (CDRs) of said VHH antibody domain.

[1226] 1999

[1227] 2000 Embodiment 224: The VHH antibody domain or fragment thereof according to any one of

[1228] 2001 embodiments 212 to 223, wherein in (F) said humanization of said sequence is by replacing at

[1229] 2002 least one amino acid of said sequence by the corresponding amino acid of a human VH (variable

[1230] 2003 heavy) domain.

[1231] 2004

[1232] 2005 Embodiment 225: The VHH antibody domain or fragment thereof according to any one of

[1233] 2006 embodiments 212 to 224, wherein in (F) said humanization of said sequence is by (individually)

[1234] 2007 replacing up to 25 amino acids of said sequence by the corresponding amino acids of a human

[1235] 2008 VH domain.

[1236] 2009

[1237] 2010 Embodiment 226: The VHH antibody domain or fragment thereof according to any one of

[1238] 2011 embodiments 212 to 224, wherein in (F) said humanization of said sequence is by replacing up

[1239] 2012 to 20 amino acids of said sequence by the corresponding amino acids of a human VH domain.

[1240] 2013 2014 Embodiment 227: The VHH antibody domain or fragment thereof according to any one of

[1241] 2015 embodiments 212 to 224, wherein in (F) said humanization of said sequence is by replacing up

[1242] 2016 to 15 amino acids of said sequence by the corresponding amino acids of a human VH domain.

[1243] 2017

[1244] 2018 Embodiment 228: The VHH antibody domain or fragment thereof according to any one of

[1245] 2019 embodiments 212 to 224, wherein in (F) said humanization of said sequence is by replacing up

[1246] 2020 to 10 amino acids of said sequence by the corresponding amino acids of a human VH domain.

[1247] 2021

[1248] 2022 Embodiment 229: The VHH antibody domain or fragment thereof according to any one of

[1249] 2023 embodiments 212 to 224, wherein in (F) said humanization of said sequence is by replacing up

[1250] 2024 to 5 amino acids of said sequence by the corresponding amino acids of a human VH domain.

[1251] 2025

[1252] 2026 Embodiment 230: The VHH antibody domain or fragment thereof according to any one of

[1253] 2027 embodiments 212 to 224, wherein in (F) said humanization of said sequence is by replacing up

[1254] 2028 to 3 amino acids of said sequence by the corresponding amino acids of a human VH domain.

[1255] 2029

[1256] 2030 Embodiment 231 : The VHH antibody domain or fragment thereof according to any one of

[1257] 2031 embodiments 212 to 224, wherein in (F) said humanization of said sequence is by replacing up

[1258] 2032 to 2 amino acids of said sequence by the corresponding amino acids of a human VH domain.

[1259] 2033

[1260] 2034 Embodiment 232: The VHH antibody domain or fragment thereof according to any one of

[1261] 2035 embodiments 212 to 224, wherein in (F) said humanization of said sequence is by replacing one

[1262] 2036 amino acid of said sequence by the corresponding amino acid of a human VH domain.

[1263] 2037

[1264] 2038 Embodiment 233: The VHH antibody domain or fragment thereof according to any one of

[1265] 2039 embodiments 212 to 232, wherein in (F) said humanization is within the framework regions of

[1266] 2040 said VHH antibody domain and / or within the CDRs of said VHH antibody domain.

[1267] 2041

[1268] 2042 Embodiment 234: The VHH antibody domain or fragment thereof according to any one of

[1269] 2043 embodiments 212 to 232, wherein in (F) said humanization is within the framework regions of

[1270] 2044 said VHH antibody domain, but not within the CDRs of said VHH antibody domain.

[1271] 2045 2046 Embodiment 235: The VHH antibody domain or fragment thereof according to any one of

[1272] 2047 embodiments 212 to 224, wherein in (F) said humanization is within the CDRs of said VHH

[1273] 2048 antibody domain, but not within the framework regions of said VHH antibody domain.

[1274] 2049

[1275] 2050 Embodiment 236: The VHH antibody domain or fragment thereof according to any one of

[1276] 2051 embodiments 212 to 233 or 235, wherein in (F) said humanization within the CDRs of said

[1277] 2052 VHH antibody domain is within CDR1, CDR2 and / or CDR3.

[1278] 2053

[1279] 2054 Embodiment 237: The VHH antibody domain or fragment thereof according to any one of

[1280] 2055 embodiments 212 to 233 or 235 to 236, wherein in (F) said humanization within the CDRs of

[1281] 2056 said VHH antibody domain is within CDR1 and / or CDR2.

[1282] 2057

[1283] 2058 Embodiment 238: The VHH antibody domain or fragment thereof according to any one of

[1284] 2059 embodiments 212 to 233 or 235 to 237, wherein in (F) said humanization within the CDRs of

[1285] 2060 said VHH antibody domain is within CDR1.

[1286] 2061

[1287] 2062 Embodiment 239: The VHH antibody domain or fragment thereof according to any one of

[1288] 2063 embodiments 212 to 233 or 235 to 238, wherein in (F) said humanization within the CDRs of

[1289] 2064 said VHH antibody domain is within CDR2.

[1290] 2065

[1291] 2066 Embodiment 240: The VHH antibody domain or fragment thereof according to any one of

[1292] 2067 embodiments 212 to 239, wherein in (F) said humanization within the CDRs of said VHH

[1293] 2068 antibody domain is not within CDR3.

[1294] 2069

[1295] 2070 Embodiment 241: The VHH antibody domain or fragment thereof according to any one of

[1296] 2071 embodiments 212 to 240, wherein in (G) the modification is the replacement, addition or

[1297] 2072 deletion of up to 20 amino acids.

[1298] 2073

[1299] 2074 Embodiment 242: The VHH antibody domain or fragment thereof according to any one of

[1300] 2075 embodiments 212 to 240, wherein in (G) the modification is the replacement, addition or

[1301] 2076 deletion of up to 15 amino acids.

[1302] 2077 2078 Embodiment 243: The VHH antibody domain or fragment thereof according to any one of

[1303] 2079 embodiments 212 to 240, wherein in (G) the modification is the replacement, addition or

[1304] 2080 deletion of up to 10 amino acids.

[1305] 2081

[1306] 2082 Embodiment 244: The VHH antibody domain or fragment thereof according to any one of

[1307] 2083 embodiments 212 to 240, wherein in (G) the modification is the replacement, addition or

[1308] 2084 deletion of up to 5 amino acids.

[1309] 2085

[1310] 2086 Embodiment 245: The VHH antibody domain or fragment thereof according to any one of

[1311] 2087 embodiments 212 to 240, wherein in (G) the modification is the replacement, addition or

[1312] 2088 deletion of up to 3 amino acids.

[1313] 2089

[1314] 2090 Embodiment 246: The VHH antibody domain or fragment thereof according to any one of

[1315] 2091 embodiments 212 to 240, wherein in (G) the modification is the replacement, addition or

[1316] 2092 deletion of up to 2 amino acids.

[1317] 2093

[1318] 2094 Embodiment 247: The VHH antibody domain or fragment thereof according to any one of

[1319] 2095 embodiments 212 to 240, wherein in (G) the modification is the replacement, addition or

[1320] 2096 deletion of one amino acid.

[1321] 2097

[1322] 2098 Embodiment 248: The VHH antibody domain or fragment thereof according to any one of

[1323] 2099 embodiments 212 to 247, wherein the modification in (G) comprises only the replacement, but

[1324] 2100 not the addition or deletion of amino acids.

[1325] 2101

[1326] 2102 Embodiment 249: The VHH antibody domain or fragment thereof according to any one of

[1327] 2103 embodiments 212 to 248, wherein in (E) said VHH antibody domain or fragment thereof

[1328] 2104 comprises the complementarity determining regions CDR1, CDR2 and CDR3 of one VHH

[1329] 2105 selected from the group consisting of VHH12, VHH13, VHH14, VHH15, VHH16, VHH17,

[1330] 2106 VHH18, VHH20, VHH21 and VHH22 as shown in the Table of CDRs.

[1331] 2107

[1332] 2108 Embodiment 250: The VHH antibody domain or fragment thereof according to any one of

[1333] 2109 embodiments 212 to 249, wherein in (F) to (H)

[1334] 2110 (d) said VHH antibody domain or fragment thereof comprises the complementarity

[1335] 2111 determining regions CDR1, CDR2 and CDR3 of one VHH selected from the group 2112 consisting of VHH12, VHH13, VHH14, VHH15, VHH16, VHH17, VHH18,

[1336] 2113 VHH20, VHH21 and VHH22 as shown in the Table of CDRs;

[1337] 2114 (e) said VHH antibody domain or fragment thereof comprises the complementarity

[1338] 2115 determining regions CDR1, CDR2 and CDR3 as defined in (d) with modification,

[1339] 2116 wherein the modification is that the sequence of at least one of CDR1, CDR2 and

[1340] 2117 CDR3 is humanized;

[1341] 2118 (f) said VHH antibody domain or fragment thereof comprises the complementarity

[1342] 2119 determining regions CDR1, CDR2 and CDR3 as defined in (d) with modification,

[1343] 2120 wherein the modification is

[1344] 2121 - the replacement, addition or deletion of up to three amino acids in CDR1,

[1345] 2122 - the replacement, addition or deletion of up to three amino acids in CDR2

[1346] 2123 and / or

[1347] 2124 - the replacement, addition or deletion of up to three amino acids in CDR3.

[1348] 2125

[1349] 2126 Embodiment 251 : The VHH antibody domain or fragment thereof according to embodiment

[1350] 2127 250, wherein the modification in (e) is that the sequence of CDR1 and / or CDR2, but not the

[1351] 2128 sequence of CDR3 is humanized.

[1352] 2129

[1353] 2130 Embodiment 252: The VHH antibody domain or fragment thereof according to any one of

[1354] 2131 embodiments 250 or 251, wherein the modification in (e) is that the sequence of CDR1 is

[1355] 2132 humanized, but not the sequence of CDR2 and CDR3.

[1356] 2133

[1357] 2134 Embodiment 253: The VHH antibody domain or fragment thereof according to any one of

[1358] 2135 embodiments 250 or 251, wherein the modification in (e) is that the sequence of CDR2 is

[1359] 2136 humanized, but not the sequence of CDR1 and CDR3.

[1360] 2137

[1361] 2138 Embodiment 254: The VHH antibody domain or fragment thereof according to any one of

[1362] 2139 embodiments 250 to 253, wherein the modification in (e) is that the sequence of one, but not

[1363] 2140 more than one of CDR1, CDR2 and CDR3 is humanized.

[1364] 2141

[1365] 2142 Embodiment 255: The VHH antibody domain or fragment thereof according to any one of

[1366] 2143 embodiments 250 to 254, wherein said humanization of said CDR(s) is by replacing at least one

[1367] 2144 amino acid in the sequence of said CDR by the corresponding amino acid of a human VH

[1368] 2145 domain. 2146

[1369] 2147 Embodiment 256: The VHH antibody domain or fragment thereof according to any one of

[1370] 2148 embodiments 250 to 255, wherein said humanization of said CDR(s) is by replacing up to three

[1371] 2149 amino acids in the sequence of said CDR by the corresponding amino acid of a human VH

[1372] 2150 domain.

[1373] 2151

[1374] 2152 Embodiment 257: The VHH antibody domain or fragment thereof according to any one of

[1375] 2153 embodiments 250 to 255, wherein said humanization of said CDR(s) is by replacing up to three

[1376] 2154 amino acids in the sequence of CDR1 and / or CDR2 and up to one amino acid in the sequence

[1377] 2155 of CDR3 by the corresponding amino acid of a human VH domain.

[1378] 2156

[1379] 2157 Embodiment 258: The VHH antibody domain or fragment thereof according to any one of

[1380] 2158 embodiments 250 to 255, wherein said humanization of said CDR(s) is by replacing up to two

[1381] 2159 amino acids in the sequence of said CDR by the corresponding amino acid of a human VH

[1382] 2160 domain.

[1383] 2161

[1384] 2162 Embodiment 259: The VHH antibody domain or fragment thereof according to any one of

[1385] 2163 embodiments 250 to 255, wherein said humanization of said CDR(s) is by replacing up to two

[1386] 2164 amino acids in the sequence of CDR1 and / or CDR2 and up to one amino acid in the sequence

[1387] 2165 of CDR3 by the corresponding amino acid of a human VH domain.

[1388] 2166

[1389] 2167 Embodiment 260: The VHH antibody domain or fragment thereof according to any one of

[1390] 2168 embodiments 250 to 259, wherein said humanization of said CDR(s) is by replacing one amino

[1391] 2169 acid in the sequence of said CDR by the corresponding amino acid of a human VH domain.

[1392] 2170

[1393] 2171 Embodiment 261 : The VHH antibody domain or fragment thereof according to any one of

[1394] 2172 embodiments 250 to 260, wherein the modification in (f) is

[1395] 2173 - the replacement, addition or deletion of up to three amino acids in CDR1,

[1396] 2174 - the replacement, addition or deletion of up to three amino acids in CDR2 and / or

[1397] 2175 - the replacement, addition or deletion of up to two amino acids in CDR3.

[1398] 2176

[1399] 2177 Embodiment 262: The VHH antibody domain or fragment thereof according to any one of

[1400] 2178 embodiments 250 to 260, wherein the modification in (f) is

[1401] 2179 - the replacement, addition or deletion of up to three amino acids in CDR1, 2180 - the replacement, addition or deletion of up to three amino acids in CDR2 and / or

[1402] 2181 - the replacement, addition or deletion of up to one amino acid in CDR3.

[1403] 2182

[1404] 2183 Embodiment 263: The VHH antibody domain or fragment thereof according to any one of

[1405] 2184 embodiments 250 to 260, wherein the modification in (f) is

[1406] 2185 - the replacement, addition or deletion of up to three amino acids in CDR1, and / or

[1407] 2186 - the replacement, addition or deletion of up to three amino acids in CDR2,

[1408] 2187 wherein the sequence of CDR3 is unmodified.

[1409] 2188

[1410] 2189 Embodiment 264: The VHH antibody domain or fragment thereof according to any one of

[1411] 2190 embodiments 250 to 260, wherein the modification in (f) is

[1412] 2191 - the replacement, addition or deletion of up to two amino acids in CDR1,

[1413] 2192 - the replacement, addition or deletion of up to two amino acids in CDR2 and / or

[1414] 2193 - the replacement, addition or deletion of up to two amino acids in CDR3;

[1415] 2194

[1416] 2195 Embodiment 265: The VHH antibody domain or fragment thereof according to any one of

[1417] 2196 embodiments 250 to 260, wherein the modification in (f) is

[1418] 2197 - the replacement, addition or deletion of up to two amino acids in CDR1;

[1419] 2198 - the replacement, addition or deletion of up to two amino acids in CDR2; and / or

[1420] 2199 - the replacement, addition or deletion of up to one amino acid in CDR3.

[1421] 2200

[1422] 2201 Embodiment 266: The VHH antibody domain or fragment thereof according to any one of

[1423] 2202 embodiments 250 to 260, wherein the modification in (f) is

[1424] 2203 - the replacement, addition or deletion of up to two amino acids in CDR1 and / or

[1425] 2204 - the replacement, addition or deletion of up to two amino acids in CDR2;

[1426] 2205 wherein the sequence of CDR3 is unmodified.

[1427] 2206

[1428] 2207 As the skilled person is aware, "unmodified" means unmodified compared to the sequence in

[1429] 2208 the Table of CDRs.

[1430] 2209

[1431] 2210 Embodiment 267: The VHH antibody domain or fragment thereof according to any one of

[1432] 2211 embodiments 250 to 260, wherein the modification in (f) is

[1433] 2212 - the replacement, addition or deletion of up to two amino acids in CDR1,

[1434] 2213 wherein the sequence of CDR2 and CDR3 is unmodified. 2214

[1435] 2215 Embodiment 268: The VHH antibody domain or fragment thereof according to any one of

[1436] 2216 embodiments 250 to 260, wherein the modification in (f) is

[1437] 2217 - the replacement, addition or deletion of up to two amino acids in CDR2,

[1438] 2218 wherein the sequence of CDR1 and CDR3 is unmodified.

[1439] 2219

[1440] 2220 Embodiment 269: The VHH antibody domain or fragment thereof according to any one of

[1441] 2221 embodiments 250 to 260, wherein the modification in (f) is

[1442] 2222 - the replacement, addition or deletion of up to one amino acid in CDR1;

[1443] 2223 - the replacement, addition or deletion of up to one amino acid in CDR2; and / or

[1444] 2224 - the replacement, addition or deletion of up to one amino acid in CDR3.

[1445] 2225

[1446] 2226 Embodiment 270: The VHH antibody domain or fragment thereof according to any one of

[1447] 2227 embodiments 250 to 260, wherein the modification in (f) is

[1448] 2228 - the replacement, addition or deletion of one amino acid in CDR1 and / or

[1449] 2229 - the replacement, addition or deletion of one amino acid in CDR2;

[1450] 2230 wherein the sequence of CDR3 is unmodified.

[1451] 2231

[1452] 2232 Embodiment 271 : The VHH antibody domain or fragment thereof according to any one of

[1453] 2233 embodiments 250 to 260, wherein the modification in (f) is

[1454] 2234 - the replacement, addition or deletion of one amino acid in CDR1,

[1455] 2235 wherein the sequence of CDR2 and CDR3 is unmodified.

[1456] 2236

[1457] 2237 Embodiment 272: The VHH antibody domain or fragment thereof according to any one of

[1458] 2238 embodiments 250 to 260, wherein the modification in (f) is

[1459] 2239 - the replacement, addition or deletion of one amino acid in CDR2,

[1460] 2240 wherein the sequence of CDR1 and CDR3 is unmodified.

[1461] 2241

[1462] 2242 Embodiment 273: The VHH antibody domain or fragment thereof according to any one of

[1463] 2243 embodiments 250 to 272, wherein the modification in (f) comprises only the replacement, but

[1464] 2244 not the addition or deletion of amino acids.

[1465] 2245

[1466] 2246 Embodiment 274: The VHH antibody domain or fragment thereof according to any one of

[1467] 2247 embodiments 250 to 273, wherein said replacement is a conservative amino acid replacement. 2248

[1468] 2249 Embodiment 275: The VHH antibody domain or fragment thereof according to any one of

[1469] 2250 embodiments 189 to 274, wherein said fragment consists of at least 100 amino acids.

[1470] 2251

[1471] 2252 Embodiment 276: The VHH antibody domain or fragment thereof according to any one of

[1472] 2253 embodiments 189 to 274, wherein said fragment consists of at least 105 amino acids.

[1473] 2254

[1474] 2255 Embodiment 277: The VHH antibody domain or fragment thereof according to any one of

[1475] 2256 embodiments 189 to 274, wherein said fragment consists of at least 110 amino acids.

[1476] 2257

[1477] 2258 Embodiment 278: The VHH antibody domain or fragment thereof according to any one of

[1478] 2259 embodiments 189 to 274, wherein said fragment consists of at least 115 amino acids.

[1479] 2260

[1480] 2261 Embodiment 279: The VHH antibody domain or fragment thereof according to any one of

[1481] 2262 embodiments 189 to 278, wherein said VHH antibody domain is an anti-IL-18RP VHH

[1482] 2263 antibody domain.

[1483] 2264

[1484] 2265 Embodiment 280: The VHH antibody domain or fragment thereof according to any one of

[1485] 2266 embodiments 189 to 279, wherein said VHH antibody domain is specific for IL-18RP.

[1486] 2267

[1487] 2268 Embodiment 281 : The VHH antibody domain or fragment thereof according to any one of

[1488] 2269 embodiments 189 to 280, wherein said VHH antibody domain or fragment thereof binds to IL-

[1489] 2270 18RP.

[1490] 2271

[1491] 2272 Embodiment 282: The VHH antibody domain or fragment thereof according to any one of

[1492] 2273 embodiments 189 to 281, wherein said VHH antibody domain or fragment thereof binds

[1493] 2274 specifically to IL-18Rp.

[1494] 2275

[1495] 2276 Embodiment 283: The VHH antibody domain or fragment thereof according to any one of

[1496] 2277 embodiments 189 to 282, wherein said VHH antibody domain or fragment thereof does not

[1497] 2278 bind to IL-18Ra.

[1498] 2279 2280 Embodiment 284: The VHH antibody domain or fragment thereof according to any one of

[1499] 2281 embodiments 189 to 283, wherein said VHH antibody domain or fragment thereof is capable

[1500] 2282 of specifically binding to IL-18RP ECD.

[1501] 2283

[1502] 2284 Embodiment 285: The VHH antibody domain or fragment thereof according to any one of

[1503] 2285 embodiments 189 to 284, wherein said VHH antibody domain or fragment thereof binds to

[1504] 2286 recombinant human IL-18RP ECD with a KD of 1x1 O'6M or stronger.

[1505] 2287

[1506] 2288 Embodiment 286 The VHH antibody domain or fragment thereof according to any one of

[1507] 2289 embodiments 189 to 284, wherein said VHH antibody domain or fragment thereof binds to

[1508] 2290 recombinant human IL-18RP ECD with a KD of 1x1 O'7M or stronger.

[1509] 2291

[1510] 2292 Embodiment 287: The VHH antibody domain or fragment thereof according to any one of

[1511] 2293 embodiments 189 to 284, wherein said VHH antibody domain or fragment thereof binds to

[1512] 2294 recombinant human IL-18RP ECD with a KD of 5x1 O'8M or stronger.

[1513] 2295

[1514] 2296 Embodiment 288: The VHH antibody domain or fragment thereof according to any one of

[1515] 2297 embodiments 189 to 284, wherein said VHH antibody domain or fragment thereof binds to

[1516] 2298 recombinant human IL-18RP ECD with a KD of 1.5x1 O'8M or stronger.

[1517] 2299

[1518] 2300 Embodiment 289: The VHH antibody domain or fragment thereof according to any one of

[1519] 2301 embodiments 189 to 284, wherein said VHH antibody domain or fragment thereof binds to

[1520] 2302 recombinant human IL-18RP ECD with a KD of 1x1 O'8M or stronger.

[1521] 2303

[1522] 2304 Embodiment 290: The VHH antibody domain or fragment thereof according to any one of

[1523] 2305 embodiments 189 to 284, wherein said VHH antibody domain or fragment thereof binds to

[1524] 2306 recombinant human IL-18RP ECD with a KD of 5x1 O'9M or stronger.

[1525] 2307

[1526] 2308 Embodiment 291 : The VHH antibody domain or fragment thereof according to any one of

[1527] 2309 embodiments 189 to 284, wherein said VHH antibody domain or fragment thereof binds to

[1528] 2310 recombinant human IL-18RP ECD with a KD of 1x1 O'9M or stronger.

[1529] 2311

[1530] 2312 Embodiment 292: The VHH antibody domain or fragment thereof according to any one of

[1531] 2313 embodiments 189 to 291, wherein said fragment of said VHH antibody domain binds to 2314 recombinant human IL-18RP ECD with at least 90% of the affinity (as determined by KD value)

[1532] 2315 with which said VHH antibody domain binds to recombinant human IL-18RP ECD.

[1533] 2316

[1534] 2317 Embodiment 293: The VHH antibody domain or fragment thereof according to any one of

[1535] 2318 embodiments 189 to 292, wherein said fragment of said VHH antibody domain binds to IL-

[1536] 2319 18Ra ECD with an affinity (as determined from the KD value) that is at least equal to the

[1537] 2320 affinity with which VHH2 (SEQ ID NO: 2) binds to IL-18Ra ECD or stronger. Such binding

[1538] 2321 can be determined by in vitro binding experiments as described in Example 1 below (by

[1539] 2322 biolayer interferometry).

[1540] 2323

[1541] 2324 Embodiment 294: The VHH antibody domain or fragment thereof according to any one of

[1542] 2325 embodiments 189 to 211 or 250 to 293, wherein in (e) and (f) the VHH antibody domain or

[1543] 2326 fragment thereof binds to recombinant human IL-18RP ECD with an affinity (KD value) that

[1544] 2327 is by not more than a factor of 10 weaker than the binding of the corresponding VHH antibody

[1545] 2328 domain without modification.

[1546] 2329

[1547] 2330 Embodiment 295: The VHH antibody domain or fragment thereof according to any one of

[1548] 2331 embodiments 189 to 211 or 250 to 293, wherein in (e) and (f) the VHH antibody domain or

[1549] 2332 fragment thereof binds to recombinant human IL-18RP ECD with an affinity (KD value) that

[1550] 2333 is by not more than a factor of 5 weaker than the binding of the corresponding VHH antibody

[1551] 2334 domain without modification.

[1552] 2335

[1553] 2336 Embodiment 296: The VHH antibody domain or fragment thereof according to any one of

[1554] 2337 embodiments 189 to 211 or 250 to 293, wherein in (e) and (f) the VHH antibody domain or

[1555] 2338 fragment thereof binds to recombinant human IL-18RP ECD with an affinity (KD value) that

[1556] 2339 is by not more than a factor of 2 weaker than the binding of the corresponding VHH antibody

[1557] 2340 domain without modification.

[1558] 2341

[1559] 2342 Embodiment 297: The VHH antibody domain or fragment thereof according to any one of

[1560] 2343 embodiments 189 to 211 or 250 to 293, wherein in (e) and (f) the VHH antibody domain or

[1561] 2344 fragment thereof binds to recombinant human IL-18RP ECD with an affinity (KD value) that

[1562] 2345 is by not more than a factor of 1.5 weaker than the binding of the corresponding VHH antibody

[1563] 2346 domain without modification.

[1564] 2347 2348 Embodiment 298: The VHH antibody domain or fragment thereof according to any one of

[1565] 2349 embodiments 189 to 211 or 250 to 297, wherein in (e) and (f) the VHH antibody domain or

[1566] 2350 fragment thereof binds to recombinant human IL-18RP ECD with an affinity (KD value) that

[1567] 2351 is by not more than a factor of 10 stronger than the binding of the corresponding VHH antibody

[1568] 2352 domain without modification.

[1569] 2353

[1570] 2354 Embodiment 299: The VHH antibody domain or fragment thereof according to any one of

[1571] 2355 embodiments 189 to 211 or 250 to 297, wherein in (e) and (f) the VHH antibody domain or

[1572] 2356 fragment thereof binds to recombinant human IL-18RP ECD with an affinity (KD value) that

[1573] 2357 is by not more than a factor of 5 stronger than the binding of the corresponding VHH antibody

[1574] 2358 domain without modification.

[1575] 2359

[1576] 2360 Embodiment 300: The VHH antibody domain or fragment thereof according to any one of

[1577] 2361 embodiments 189 to 211 or 250 to 297, wherein in (e) and (f) the VHH antibody domain or

[1578] 2362 fragment thereof binds to recombinant human IL-18RP ECD with an affinity (KD value) that

[1579] 2363 is by not more than a factor of 2 stronger than the binding of the corresponding VHH antibody

[1580] 2364 domain without modification.

[1581] 2365

[1582] 2366 Embodiment 301 : The VHH antibody domain or fragment thereof according to any one of

[1583] 2367 embodiments 189 to 211 or 250 to 297, wherein in (e) and (f) the VHH antibody domain or

[1584] 2368 fragment thereof binds to recombinant human IL-18RP ECD with an affinity (KD value) that

[1585] 2369 is by not more than a factor of 1.5 stronger than the binding of the corresponding VHH antibody

[1586] 2370 domain without modification.

[1587] 2371

[1588] 2372 Embodiment 302: The VHH antibody domain or fragment thereof according to any one of

[1589] 2373 embodiments 212 to 301, wherein in (F) and (G) the VHH antibody domain binds to

[1590] 2374 recombinant human IL-18RP ECD with an affinity (KD value) that is by not more than a factor

[1591] 2375 of 10 weaker than the binding of the corresponding VHH antibody domain without

[1592] 2376 modification.

[1593] 2377

[1594] 2378 Embodiment 303: The VHH antibody domain or fragment thereof according to any one of

[1595] 2379 embodiments 212 to 301, wherein in (F) and (G) the VHH antibody domain binds to

[1596] 2380 recombinant human IL-18RP ECD with an affinity (KD value) that is by not more than a factor

[1597] 2381 of 5 weaker than the binding of the corresponding VHH antibody domain without modification. 2382

[1598] 2383 Embodiment 304: The VHH antibody domain or fragment thereof according to any one of

[1599] 2384 embodiments 212 to 301, wherein in (F) and (G) the VHH antibody domain binds to

[1600] 2385 recombinant human IL-18RP ECD with an affinity (KD value) that is by not more than a factor

[1601] 2386 of 2 weaker than the binding of the corresponding VHH antibody domain without modification.

[1602] 2387

[1603] 2388 Embodiment 305: The VHH antibody domain or fragment thereof according to any one of

[1604] 2389 embodiments 189 to 301, wherein in (F) and (G) the VHH antibody domain binds to

[1605] 2390 recombinant human IL-18RP ECD with an affinity (KD value) that is by not more than a factor

[1606] 2391 of 1.5 weaker than the binding of the corresponding VHH antibody domain without

[1607] 2392 modification.

[1608] 2393

[1609] 2394 Embodiment 306: The VHH antibody domain or fragment thereof according to any one of

[1610] 2395 embodiments 212 to 305, wherein in (F) and (G) the VHH antibody domain binds to

[1611] 2396 recombinant human IL-18RP ECD with an affinity (KD value) that is by not more than a factor

[1612] 2397 of 10 stronger than the binding of the corresponding VHH antibody domain without

[1613] 2398 modification.

[1614] 2399

[1615] 2400 Embodiment 307: The VHH antibody domain or fragment thereof according to any one of

[1616] 2401 embodiments 212 to 305, wherein in (F) and (G) the VHH antibody domain binds to

[1617] 2402 recombinant human IL-18RP ECD with an affinity (KD value) that is by not more than a factor

[1618] 2403 of 5 stronger than the binding of the corresponding VHH antibody domain without

[1619] 2404 modification.

[1620] 2405

[1621] 2406 Embodiment 308: The VHH antibody domain or fragment thereof according to any one of

[1622] 2407 embodiments 212 to 305, wherein in (F) and (G) the VHH antibody domain binds to

[1623] 2408 recombinant human IL-18RP ECD with an affinity (KD value) that is by not more than a factor

[1624] 2409 of 2 stronger than the binding of the corresponding VHH antibody domain without

[1625] 2410 modification.

[1626] 2411

[1627] 2412 Embodiment 309: The VHH antibody domain or fragment thereof according to any one of

[1628] 2413 embodiments 212 to 305, wherein in (F) and (G) the VHH antibody domain binds to

[1629] 2414 recombinant human IL-18RP ECD with an affinity (KD value) that is by not more than a factor 2415 of 1.5 stronger than the binding of the corresponding VHH antibody domain without

[1630] 2416 modification.

[1631] 2417

[1632] 2418 Embodiment 310: The VHH antibody domain or fragment thereof according to any one of

[1633] 2419 embodiments 212 to 309, wherein in (H) the affinity (KD value) of the binding of the VHH

[1634] 2420 antibody domain to recombinant human IL-18RP ECD is by not more than a factor of 10 weaker

[1635] 2421 than the affinity (KD value) of the binding to recombinant human IL-18RP ECD of a VHH

[1636] 2422 antibody domain consisting of the sequence from the Table of VHH Sequences that has the

[1637] 2423 highest degree of sequence identity with the sequence of said VHH antibody domain of (H).

[1638] 2424 The degree of sequence identity can be determined by sequence alignment.

[1639] 2425

[1640] 2426 Embodiment 311 : The VHH antibody domain or fragment thereof according to any one of

[1641] 2427 embodiments 212 to 309, wherein in (H) the affinity (KD value) of the binding of the VHH

[1642] 2428 antibody domain to recombinant human IL-18RP ECD is by not more than a factor of 5 weaker

[1643] 2429 than the affinity (KD value) of the binding to recombinant human IL-18RP ECD of a VHH

[1644] 2430 antibody domain consisting of the sequence from the Table of VHH Sequences that has the

[1645] 2431 highest degree of sequence identity with the sequence of said VHH antibody domain of (H).

[1646] 2432 The degree of sequence identity can be determined by sequence alignment.

[1647] 2433

[1648] 2434 Embodiment 312: The VHH antibody domain or fragment thereof according to any one of

[1649] 2435 embodiments 212 to 309, wherein in (H) the affinity (KD value) of the binding of the VHH

[1650] 2436 antibody domain to recombinant human IL-18RP ECD is by not more than a factor of 2 weaker

[1651] 2437 than the affinity (KD value) of the binding to recombinant human IL-18RP ECD of a VHH

[1652] 2438 antibody domain consisting of the sequence from the Table of VHH Sequences that has the

[1653] 2439 highest degree of sequence identity with the sequence of said VHH antibody domain of (H).

[1654] 2440

[1655] 2441 Embodiment 313: The VHH antibody domain or fragment thereof according to any one of

[1656] 2442 embodiments 212 to 309, wherein in (H) the affinity (KD value) of the binding of the VHH

[1657] 2443 antibody domain to recombinant human IL-18RP ECD is by not more than a factor of 1.5

[1658] 2444 weaker than the affinity (KD value) of the binding to recombinant human IL-18RP ECD of a

[1659] 2445 VHH antibody domain consisting of the sequence from the Table of VHH Sequences that has

[1660] 2446 the highest degree of sequence identity with the sequence of said VHH antibody domain of (H).

[1661] 2447 2448 Embodiment 314 The VHH antibody domain or fragment thereof according to any one of

[1662] 2449 embodiments 212 to 313, wherein in (H) the affinity (KD value) of the binding of the VHH

[1663] 2450 antibody domain to recombinant human IL-18RP ECD is by not more than a factor of 10

[1664] 2451 stronger than the affinity (KD value) of the binding to recombinant human IL-18RP ECD of a

[1665] 2452 VHH antibody domain consisting of the sequence from the Table of VHH Sequences that has

[1666] 2453 the highest degree of sequence identity with the sequence of said VHH antibody domain of (H).

[1667] 2454

[1668] 2455 Embodiment 315: The VHH antibody domain or fragment thereof according to any one of

[1669] 2456 embodiments 212 to 313, wherein in (H) the affinity (KD value) of the binding of the VHH

[1670] 2457 antibody domain to recombinant human IL-18RP ECD is by not more than a factor of 5 stronger

[1671] 2458 than the affinity (KD value) of the binding to recombinant human IL-18RP ECD of a VHH

[1672] 2459 antibody domain consisting of the sequence from the Table of VHH Sequences that has the

[1673] 2460 highest degree of sequence identity with the sequence of said VHH antibody domain of (H).

[1674] 2461

[1675] 2462 Embodiment 316: The VHH antibody domain or fragment thereof according to any one of

[1676] 2463 embodiments 212 to 313, wherein in (H) the affinity (KD value) of the binding of the VHH

[1677] 2464 antibody domain to recombinant human IL-18RP ECD is by not more than a factor of 2 stronger

[1678] 2465 than the affinity (KD value) of the binding to recombinant human IL-18RP ECD of a VHH

[1679] 2466 antibody domain consisting of the sequence from the Table of VHH Sequences that has the

[1680] 2467 highest degree of sequence identity with the sequence of said VHH antibody domain of (H).

[1681] 2468

[1682] 2469 Embodiment 317: The VHH antibody domain or fragment thereof according to any one of

[1683] 2470 embodiments 212 to 313, wherein in (H) the affinity (KD value) of the binding of the VHH

[1684] 2471 antibody domain to recombinant human IL-18RP ECD is by not more than a factor of 1.5

[1685] 2472 stronger than the affinity (KD value) of the binding to recombinant human IL-18RP ECD of a

[1686] 2473 VHH antibody domain consisting of the sequence from the Table of VHH Sequences that has

[1687] 2474 the highest degree of sequence identity with the sequence of said VHH antibody domain of (H).

[1688] 2475

[1689] 2476 Embodiment 318: The VHH antibody domain or fragment thereof according to any one of

[1690] 2477 embodiments 279 to 317, wherein said KD value is determined in in vitro binding experiments

[1691] 2478 through biolayer interferometry.

[1692] 2479

[1693] 2480 Embodiment 319: The VHH antibody domain or fragment thereof according to any one of

[1694] 2481 embodiments 279 to 318, wherein said KD value is measured by kinetic measurements by 2482 biolayer interferometry at 25°C and 1000 rpm in KB Buffer (PBS + 0.1 % Tween-20 +

[1695] 2483 1% BSA).

[1696] 2484

[1697] 2485 Embodiment 320: The VHH antibody domain or fragment thereof according to any one of

[1698] 2486 embodiments 212 to 319, wherein said VHH antibody domain of (E) comprises one VHH

[1699] 2487 sequence selected from the group consisting of VHH12, VHH13, VHH14, VHH15, VHH17,

[1700] 2488 VHH18, VHH20, VHH21 and VHH22 shown in the Table of VHH Sequences.

[1701] 2489

[1702] 2490 Embodiment 321 : The VHH antibody domain or fragment thereof according to any one of

[1703] 2491 embodiments 212 to 319, wherein said VHH antibody domain of (E) comprises one VHH

[1704] 2492 sequence selected from the group consisting of VHH12, VHH14, VHH15, VHH17 and VHH22

[1705] 2493 shown in the Table of VHH Sequences.

[1706] 2494

[1707] 2495 Embodiment 322: The VHH antibody domain or fragment thereof according to any one of

[1708] 2496 embodiments 212 to 319, wherein said VHH antibody domain of (E) comprises one VHH

[1709] 2497 sequence selected from the group consisting of VHH6 and VHH11 shown in the Table of VHH

[1710] 2498 Sequences.

[1711] 2499

[1712] 2500 Embodiment 323: The VHH antibody domain or fragment thereof according to any one of

[1713] 2501 embodiments 212 to 319, wherein said VHH antibody domain of (E) comprises one VHH

[1714] 2502 sequence selected from the group consisting of VHH13, VHH16 and VHH22 shown in the

[1715] 2503 Table of VHH Sequences.

[1716] 2504

[1717] 2505 Embodiment 324: The VHH antibody domain or fragment thereof according to any one of

[1718] 2506 embodiments 212 to 319, wherein said VHH antibody domain of (E) comprises one VHH

[1719] 2507 sequence selected from the group consisting of VHH12, VHH13, VHH15, VHH17, VHH18

[1720] 2508 and VHH21 shown in the Table of VHH Sequences.

[1721] 2509

[1722] 2510 Embodiment 325: The VHH antibody domain or fragment thereof according to any one of

[1723] 2511 embodiments 212 to 319, wherein said VHH antibody domain of (E) comprises one VHH

[1724] 2512 sequence selected from the group consisting of VHH15 and VHH17 shown in the Table of

[1725] 2513 VHH Sequences.

[1726] 2514 2515 Embodiment 326: The VHH antibody domain or fragment thereof according to any one of

[1727] 2516 embodiments 212 to 319, wherein said VHH antibody domain of (E) comprises the VHH

[1728] 2517 sequence VHH12 shown in the Table of VHH Sequences.

[1729] 2518

[1730] 2519 Embodiment 327: The VHH antibody domain or fragment thereof according to any one of

[1731] 2520 embodiments 212 to 319, wherein said VHH antibody domain of (E) comprises the VHH

[1732] 2521 sequence VHH13 shown in the Table of VHH Sequences.

[1733] 2522

[1734] 2523 Embodiment 328: The VHH antibody domain or fragment thereof according to any one of

[1735] 2524 embodiments 212 to 319, wherein said VHH antibody domain of (E) comprises the VHH

[1736] 2525 sequence VHH14 shown in the Table of VHH Sequences.

[1737] 2526

[1738] 2527 Embodiment 329: The VHH antibody domain or fragment thereof according to any one of

[1739] 2528 embodiments 212 to 319, wherein said VHH antibody domain of (E) comprises the VHH

[1740] 2529 sequence VHH15 shown in the Table of VHH Sequences.

[1741] 2530

[1742] 2531 Embodiment 330: The VHH antibody domain or fragment thereof according to any one of

[1743] 2532 embodiments 212 to 319, wherein said VHH antibody domain of (E) comprises the VHH

[1744] 2533 sequence VHH16 shown in the Table of VHH Sequences.

[1745] 2534

[1746] 2535 Embodiment 331 : The VHH antibody domain or fragment thereof according to any one of

[1747] 2536 embodiments 212 to 319, wherein said VHH antibody domain of (E) comprises the VHH

[1748] 2537 sequence VHH17 shown in the Table of VHH Sequences.

[1749] 2538

[1750] 2539 Embodiment 332: The VHH antibody domain or fragment thereof according to any one of

[1751] 2540 embodiments 212 to 319, wherein said VHH antibody domain of (E) comprises the VHH

[1752] 2541 sequence VHH18 shown in the Table of VHH Sequences.

[1753] 2542

[1754] 2543 Embodiment 333: The VHH antibody domain or fragment thereof according to any one of

[1755] 2544 embodiments 212 to 319, wherein said VHH antibody domain of (E) comprises the VHH

[1756] 2545 sequence VHH20 shown in the Table of VHH Sequences.

[1757] 2546 2547 Embodiment 334: The VHH antibody domain or fragment thereof according to any one of

[1758] 2548 embodiments 212 to 319, wherein said VHH antibody domain of (E) comprises the VHH

[1759] 2549 sequence VHH21 shown in the Table of VHH Sequences.

[1760] 2550

[1761] 2551 Embodiment 335: The VHH antibody domain or fragment thereof according to any one of

[1762] 2552 embodiments 212 to 319, wherein said VHH antibody domain of (E) comprises the VHH

[1763] 2553 sequence VHH22 shown in the Table of VHH Sequences.

[1764] 2554

[1765] 2555 Embodiment 336: The VHH antibody domain or fragment thereof according to any one of

[1766] 2556 embodiments 189 to 211 or 250 to 335, wherein in (d) said VHH antibody domain or fragment

[1767] 2557 thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of one

[1768] 2558 VHH selected from the group consisting of VHH12, VHH13, VHH14, VHH15, VHH17,

[1769] 2559 VHH18, VHH20, VHH21 or VHH22 as shown in the Table of CDRs.

[1770] 2560

[1771] 2561 Embodiment 337: The VHH antibody domain or fragment thereof according to any one of

[1772] 2562 embodiments 189 to 211 or 250 to 335, wherein in (d) said VHH antibody domain or fragment

[1773] 2563 thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of one

[1774] 2564 VHH selected from the group consisting of VHH12, VHH14, VHH15, VHH17 or VHH22 as

[1775] 2565 shown in the Table of CDRs.

[1776] 2566

[1777] 2567 Embodiment 338: The VHH antibody domain or fragment thereof according to any one of

[1778] 2568 embodiments 189 to 211 or 250 to 335, wherein in (d) said VHH antibody domain or fragment

[1779] 2569 thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of VHH6

[1780] 2570 or VHH11 as shown in the Table of CDRs.

[1781] 2571

[1782] 2572 Embodiment 339: The VHH antibody domain or fragment thereof according to any one of

[1783] 2573 embodiments 189 to 211 or 250 to 335, wherein in (d) said VHH antibody domain or fragment

[1784] 2574 thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of one

[1785] 2575 VHH selected from the group consisting of VHH13, VHH16 or VHH22 as shown in the Table

[1786] 2576 of CDRs.

[1787] 2577

[1788] 2578 Embodiment 340: The VHH antibody domain or fragment thereof according to any one of

[1789] 2579 embodiments 189 to 211 or 250 to 335, wherein in (d) said VHH antibody domain or fragment

[1790] 2580 thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of one 2581 VHH selected from the group consisting of VHH 12, VHH13, VHH15, VHH17, VHH18 or

[1791] 2582 VHH21 as shown in the Table of CDRs.

[1792] 2583

[1793] 2584 Embodiment 341 : The VHH antibody domain or fragment thereof according to any one of

[1794] 2585 embodiments 189 to 211 or 250 to 335, wherein in (d) said VHH antibody domain or fragment

[1795] 2586 thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of one

[1796] 2587 VHH selected from the group consisting of VHH15 or VHH17 as shown in the Table of CDRs.

[1797] 2588

[1798] 2589 Embodiment 342: The VHH antibody domain or fragment thereof according to any one of

[1799] 2590 embodiments 189 to 211 or 250 to 335, wherein in (d) said VHH antibody domain or fragment

[1800] 2591 thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of

[1801] 2592 VHH12 as shown in the Table of CDRs.

[1802] 2593

[1803] 2594 Embodiment 343: The VHH antibody domain or fragment thereof according to any one of

[1804] 2595 embodiments 189 to 211 or 250 to 335, wherein in (d) said VHH antibody domain or fragment

[1805] 2596 thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of

[1806] 2597 VHH13 as shown in the Table of CDRs.

[1807] 2598

[1808] 2599 Embodiment 344: The VHH antibody domain or fragment thereof according to any one of

[1809] 2600 embodiments 189 to 211 or 250 to 335, wherein in (d) said VHH antibody domain or fragment

[1810] 2601 thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of

[1811] 2602 VHH14 as shown in the Table of CDRs.

[1812] 2603

[1813] 2604 Embodiment 345: The VHH antibody domain or fragment thereof according to any one of

[1814] 2605 embodiments 189 to 211 or 250 to 335, wherein in (d) said VHH antibody domain or fragment

[1815] 2606 thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of

[1816] 2607 VHH15 as shown in the Table of CDRs.

[1817] 2608

[1818] 2609 Embodiment 346: The VHH antibody domain or fragment thereof according to any one of

[1819] 2610 embodiments 189 to 211 or 250 to 335, wherein in (d) said VHH antibody domain or fragment

[1820] 2611 thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of

[1821] 2612 VHH16 as shown in the Table of CDRs.

[1822] 2613 2614 Embodiment 347: The VHH antibody domain or fragment thereof according to any one of

[1823] 2615 embodiments 189 to 211 or 250 to 335, wherein in (d) said VHH antibody domain or fragment

[1824] 2616 thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of

[1825] 2617 VHH17 as shown in the Table of CDRs.

[1826] 2618

[1827] 2619 Embodiment 348: The VHH antibody domain or fragment thereof according to any one of

[1828] 2620 embodiments 189 to 211 or 250 to 335, wherein in (d) said VHH antibody domain or fragment

[1829] 2621 thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of

[1830] 2622 VHH18 as shown in the Table of CDRs.

[1831] 2623

[1832] 2624 Embodiment 349: The VHH antibody domain or fragment thereof according to any one of

[1833] 2625 embodiments 189 to 211 or 250 to 335, wherein in (d) said VHH antibody domain or fragment

[1834] 2626 thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of

[1835] 2627 VHH20 as shown in the Table of CDRs.

[1836] 2628

[1837] 2629 Embodiment 350: The VHH antibody domain or fragment thereof according to any one of

[1838] 2630 embodiments 189 to 211 or 250 to 335, wherein in (d) said VHH antibody domain or fragment

[1839] 2631 thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of

[1840] 2632 VHH21 as shown in the Table of CDRs.

[1841] 2633

[1842] 2634 Embodiment 351: The VHH antibody domain or fragment thereof according to any one of

[1843] 2635 embodiments 189 to 211 or 250 to 335, wherein in (d) said VHH antibody domain or fragment

[1844] 2636 thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of

[1845] 2637 VHH22 as shown in the Table of CDRs.

[1846] 2638

[1847] 2639 Embodiment 352: The VHH antibody domain or fragment thereof according to any one of

[1848] 2640 embodiments 189 to 351, wherein said VHH antibody domain or fragment thereof competes

[1849] 2641 with VHH15 for binding to recombinant human IL-18RP ECD.

[1850] 2642

[1851] 2643 Embodiment 353: The VHH antibody domain or fragment thereof according to any one of

[1852] 2644 embodiments 189 to 351, wherein said VHH antibody domain or fragment thereof partially

[1853] 2645 competes with VHH15 for binding to recombinant human IL-18RP ECD.

[1854] 2646 2647 Embodiment 354: The VHH antibody domain or fragment thereof according to any one of

[1855] 2648 embodiments 189 to 353, wherein said VHH antibody domain or fragment thereof does not

[1856] 2649 compete with VHH2 for binding to recombinant human IL-18Ra ECD.

[1857] 2650

[1858] 2651 Embodiment 355: The VHH antibody domain or fragment thereof according to any one of

[1859] 2652 embodiments 189 to 354, wherein said humanization is by germlining.

[1860] 2653

[1861] 2654 Embodiment 356: The VHH antibody domain or fragment thereof according to embodiment

[1862] 2655 355, wherein said germlining involves

[1863] 2656 - identifying the human VH germline sequence that is closest to the sequence to be

[1864] 2657 humanized with respect to its sequence similarity; and

[1865] 2658 - replacing amino acid(s) in the sequence to be humanized by the amino acid(s) at the

[1866] 2659 corresponding sequence position(s) in the closest human VH germline sequence.

[1867] 2660

[1868] 2661 Sequence similarity and corresponding sequence positions can be determined by sequence

[1869] 2662 alignment as described above.

[1870] 2663

[1871] 2664 Embodiment 357: The VHH antibody domain or fragment thereof according to embodiment

[1872] 2665 355, wherein said germlining consists of

[1873] 2666 - identifying the human VH germline sequence that is closest to the sequence to be

[1874] 2667 humanized with respect to its sequence similarity; and

[1875] 2668 - replacing amino acid(s) in the sequence to be humanized by the amino acid(s) at the

[1876] 2669 corresponding sequence position(s) in the closest human VH germline sequence.

[1877] 2670

[1878] 2671 Embodiment 358: The VHH antibody domain or fragment thereof according to any one of

[1879] 2672 embodiments 189 to 357, wherein said VHH antibody domain (resp. fragment thereof) with

[1880] 2673 humanization binds to human IL-18RP ECD with an affinity (KD value) that is not weaker by

[1881] 2674 a factor of more than 10 compared to the binding of a corresponding VHH antibody domain

[1882] 2675 (resp. fragment thereof) without humanization to human IL-18RP ECD.

[1883] 2676

[1884] 2677 As a skilled person understands, the term "corresponding VHH antibody domain without

[1885] 2678 humanization" refers to a VHH antibody domain that differs only by the amino acid changes

[1886] 2679 introduced in order to carry out said humanization, but that is otherwise identical.

[1887] 2680 2681 Embodiment 359: The VHH antibody domain or fragment thereof according to any one of

[1888] 2682 embodiments 189 to 357, wherein said VHH antibody domain (resp. fragment thereof) with

[1889] 2683 humanization binds to human IL-18RP ECD with an affinity (KD value) that is not weaker by

[1890] 2684 a factor of more than 5 compared to the binding of a corresponding VHH antibody domain

[1891] 2685 (resp. fragment thereof) without humanization to human IL-18RP ECD.

[1892] 2686

[1893] 2687 Embodiment 360: The VHH antibody domain or fragment thereof according to any one of

[1894] 2688 embodiments 189 to 357, wherein said VHH antibody domain (resp. fragment thereof) with

[1895] 2689 humanization binds to human IL-18RP ECD with an affinity (KD value) that is not weaker by

[1896] 2690 a factor of more than 2 compared to the binding of a corresponding VHH antibody domain

[1897] 2691 (resp. fragment thereof) without humanization to human IL-18RP ECD.

[1898] 2692

[1899] 2693 Embodiment 361 : The VHH antibody domain or fragment thereof according to any one of

[1900] 2694 embodiments 189 to 357, wherein said VHH antibody domain (resp. fragment thereof) with

[1901] 2695 humanization binds to human IL-18RP ECD with an affinity (KD value) that is not weaker than

[1902] 2696 the binding of a corresponding VHH antibody domain (resp. fragment thereof) without

[1903] 2697 humanization to human IL-18RP ECD.

[1904] 2698

[1905] 2699 Embodiment 362: The VHH antibody domain or fragment thereof according to any one of

[1906] 2700 embodiments 358 to 361, wherein said affinity is determined by KD measurement.

[1907] 2701

[1908] 2702 Embodiment 363: The VHH antibody domain or fragment thereof according to any one of

[1909] 2703 embodiments 212 to 362, wherein said VHH antibody domain comprises a VHH antibody

[1910] 2704 domain according to (E), (F) or (G) / said fragment of a VHH antibody domain comprises a

[1911] 2705 fragment of a VHH antibody domain according to (E), (F) or (G).

[1912] 2706

[1913] 2707 Embodiment 364: The VHH antibody domain or fragment thereof according to any one of

[1914] 2708 embodiments 212 to 362, wherein said VHH antibody domain comprises a VHH antibody

[1915] 2709 domain according to (E), (F) or (H) / said fragment of a VHH antibody domain comprises a

[1916] 2710 fragment of a VHH antibody domain according to (E), (F) or (H).

[1917] 2711

[1918] 2712 Embodiment 365: The VHH antibody domain or fragment thereof according to any one of

[1919] 2713 embodiments 212 to 362, wherein said VHH antibody domain comprises a VHH antibody 2714 domain according to (E), (G) or (H) / said fragment of a VHH antibody domain comprises a

[1920] 2715 fragment of a VHH antibody domain according to (E), (G) or (H).

[1921] 2716

[1922] 2717 Embodiment 366: The VHH antibody domain or fragment thereof according to any one of

[1923] 2718 embodiments 212 to 362, wherein said VHH antibody domain comprises a VHH antibody

[1924] 2719 domain according to (E) or (F) / said fragment of a VHH antibody domain comprises a fragment

[1925] 2720 of a VHH antibody domain according to (E) or (F).

[1926] 2721

[1927] 2722 Embodiment 367: The VHH antibody domain or fragment thereof according to any one of

[1928] 2723 embodiments 212 to 362, wherein said VHH antibody domain comprises a VHH antibody

[1929] 2724 domain according to (E) or (G) / said fragment of a VHH antibody domain comprises a fragment

[1930] 2725 of a VHH antibody domain according to (E) or (G).

[1931] 2726 1 1 Embodiment 368: The VHH antibody domain or fragment thereof according to any one of

[1932] 2728 embodiments 212 to 362, wherein said VHH antibody domain comprises a VHH antibody

[1933] 2729 domain according to (E) or (H) / said fragment of a VHH antibody domain comprises a fragment

[1934] 2730 of a VHH antibody domain according to (E) or (H).

[1935] 2731

[1936] 2732 Embodiment 369: The VHH antibody domain or fragment thereof according to any one of

[1937] 2733 embodiments 212 to 362, wherein said VHH antibody domain comprises a VHH antibody

[1938] 2734 domain according to (E) / said fragment of a VHH antibody domain comprises a fragment of a

[1939] 2735 VHH antibody domain according to (E).

[1940] 2736

[1941] 2737 Embodiment 370: The VHH antibody domain or fragment thereof according to any one of

[1942] 2738 embodiments 212 to 362, wherein said VHH antibody domain comprises a VHH antibody

[1943] 2739 domain according to (F) / said fragment of a VHH antibody domain comprises a fragment of a

[1944] 2740 VHH antibody domain according to (F).

[1945] 2741

[1946] 2742 Embodiment 371 : The VHH antibody domain or fragment thereof according to any one of

[1947] 2743 embodiments 189 to 211 or 250 to 370, wherein said VHH antibody domain or fragment thereof

[1948] 2744 comprises complementarity determining regions according to (d) or (e).

[1949] 2745 2746 Embodiment 372: The VHH antibody domain or fragment thereof according to any one of

[1950] 2747 embodiments 189 to 211 or 250 to 370, wherein said VHH antibody domain or fragment thereof

[1951] 2748 comprises complementarity determining regions according to (d) or (f).

[1952] 2749

[1953] 2750 Embodiment 373: The VHH antibody domain or fragment thereof according to any one of

[1954] 2751 embodiments 189 to 211 or 250 to 370, wherein said VHH antibody domain or fragment thereof

[1955] 2752 comprises complementarity determining regions according to (d).

[1956] 2753

[1957] 2754 Embodiment 374: The VHH antibody domain or fragment thereof according to any one of

[1958] 2755 embodiments 189 to 211 or 250 to 370, wherein said VHH antibody domain or fragment thereof

[1959] 2756 comprises complementarity determining regions according to (e).

[1960] 2757

[1961] 2758 Seventh aspect of the present disclosure (also referred to as "Embodiment 375"): According to

[1962] 2759 a seventh aspect, the present disclosure relates to a compound comprising

[1963] 2760 - a VHH antibody domain or fragment thereof according to any one of embodiments 1 to

[1964] 2761 188.

[1965] 2762

[1966] 2763 Eighth aspect of the present disclosure (also referred to as "Embodiment 376"): According to a

[1967] 2764 eighth aspect, the present disclosure relates to a compound comprising

[1968] 2765 - a VHH antibody domain or fragment thereof according to any one of claims 189 to 374.

[1969] 2766

[1970] 2767 Nineth aspect of the present disclosure (also referred to as "Embodiment 377"): According to a

[1971] 2768 nineth aspect, the present disclosure relates to a compound comprising

[1972] 2769 - a first binding module which is a VHH antibody domain or fragment thereof, wherein

[1973] 2770 (a) said VHH antibody domain or fragment thereof comprises the

[1974] 2771 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH

[1975] 2772 selected from the group consisting of VHH1, VHH2, VHH3, VHH4, VHH5,

[1976] 2773 VHH6, VHH8, VHH9, VHH10 and VHH11 as shown in the Table of CDR;

[1977] 2774 (b) said VHH antibody domain or fragment thereof comprises the

[1978] 2775 complementarity determining regions CDR1, CDR2 and CDR3 as defined in

[1979] 2776 (a) with modification, wherein the modification is that the sequence of at least

[1980] 2777 one of CDR1, CDR2 and CDR3 is humanized; or 2778 (c) said VHH antibody domain or fragment thereof comprises the

[1981] 2779 complementarity determining regions CDR1, CDR2 and CDR3 as defined in

[1982] 2780 (a) with modification, wherein the modification is

[1983] 2781 - the replacement, addition or deletion of up to three amino acids in

[1984] 2782 CDR1,

[1985] 2783 - the replacement, addition or deletion of up to three amino acids in

[1986] 2784 CDR2 and / or

[1987] 2785 - the replacement, addition or deletion of up to three amino acids in

[1988] 2786 CDR3;

[1989] 2787 - a second binding module which is a VHH antibody domain or a fragment thereof, wherein

[1990] 2788 (d) said VHH antibody domain or fragment thereof comprises the

[1991] 2789 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH

[1992] 2790 selected from the group consisting of VHH12, VHH13, VHH14, VHH15,

[1993] 2791 VHH16, VHH17, VHH18, VHH20, VHH21 and VHH22 as shown in the

[1994] 2792 Table of CDRs;

[1995] 2793 (e) said VHH antibody domain or fragment thereof comprises the

[1996] 2794 complementarity determining regions CDR1, CDR2 and CDR3 as defined in

[1997] 2795 (d) with modification, wherein the modification is that the sequence of at least

[1998] 2796 one of CDR1, CDR2 and CDR3 is humanized; or

[1999] 2797 (f) said VHH antibody domain or fragment thereof comprises the

[2000] 2798 complementarity determining regions CDR1, CDR2 and CDR3 as defined in

[2001] 2799 (d) with modification, wherein the modification is

[2002] 2800 - the replacement, addition or deletion of up to three amino acids in

[2003] 2801 CDR1,

[2004] 2802 - the replacement, addition or deletion of up to three amino acids in

[2005] 2803 CDR2 and / or

[2006] 2804 - the replacement, addition or deletion of up to three amino acids in

[2007] 2805 CDR3.

[2008] 2806

[2009] 2807 To the seventh to nineth aspect of the present disclosure and the embodiments referring thereto,

[2010] 2808 the same explanations and definitions apply accordingly as for the first to sixth aspects of the

[2011] 2809 present disclosure and the embodiments referring thereto.

[2012] 2810 2811 For embodiment 377 and embodiments referring thereto, with respect to said first binding

[2013] 2812 module further embodiments of said VHH antibody domain or fragment thereof of said first

[2014] 2813 binding module are as defined in embodiments 1 to 188.

[2015] 2814

[2016] 2815 For embodiment 377 and embodiments referring thereto, with respect to said second binding

[2017] 2816 module further embodiments of said VHH antibody domain or fragment thereof of said second

[2018] 2817 binding module are as defined in embodiments 189 to 374.

[2019] 2818

[2020] 2819 Tenth aspect of the present disclosure (also referred to as "Embodiment 378"): According to a

[2021] 2820 tenth aspect, the present disclosure relates to a compound comprising

[2022] 2821 - a first binding module which is a VHH antibody domain or fragment thereof, wherein

[2023] 2822 (A) said VHH antibody domain comprises the amino acid sequence of a VHH

[2024] 2823 selected from the group consisting of VHH1, VHH2, VHH3, VHH4, VHH5,

[2025] 2824 VHH6, VHH8, VHH9, VHH10 and VHH11 as shown in the Table of VHH

[2026] 2825 Sequences;

[2027] 2826 (B) said VHH antibody domain comprises a VHH sequence as defined in (A) with

[2028] 2827 modification, wherein the modification is that said sequence is humanized;

[2029] 2828 (C) said VHH antibody domain comprises a VHH sequence as defined in (A) with

[2030] 2829 modification, wherein the modification is the replacement, addition or

[2031] 2830 deletion of up to 25 amino acids; or

[2032] 2831 (D) said VHH antibody domain comprises a VHH sequence that is at least 75%

[2033] 2832 identical to a VHH sequence referred to in (A);

[2034] 2833 - a second binding module which is a VHH antibody domain or a fragment thereof, wherein

[2035] 2834 (E) said VHH antibody domain comprises the amino acid sequence of a VHH

[2036] 2835 selected from the group consisting of VHH12, VHH13, VHH14, VHH15,

[2037] 2836 VHH16, VHH17, VHH18, VHH20, VHH21 and VHH22 as shown in the

[2038] 2837 Table of VHH Sequences;

[2039] 2838 (F) said VHH antibody domain comprises a VHH sequence as defined in (E) with

[2040] 2839 modification, wherein the modification is that said sequence is humanized;

[2041] 2840 (G) said VHH antibody domain comprises a VHH sequence as defined in (E) with

[2042] 2841 modification, wherein the modification is the replacement, addition or

[2043] 2842 deletion of up to 25 amino acids; or

[2044] 2843 (H) said VHH antibody domain comprises a VHH sequence that is at least 75%

[2045] 2844 identical to a VHH sequence referred to in (E). 2845

[2046] 2846 To the tenth aspect of the present disclosure and the embodiments referring thereto, the same

[2047] 2847 explanations and definitions apply accordingly as for the first to nineth aspects of the present

[2048] 2848 disclosure and the embodiments referring thereto.

[2049] 2849

[2050] 2850 For embodiment 378 and embodiments referring thereto, with respect to said first binding

[2051] 2851 module further embodiments of said VHH antibody domain or fragment thereof of said first

[2052] 2852 binding module are as defined in embodiments 1 to 188.

[2053] 2853

[2054] 2854 For embodiment 378 and embodiments referring thereto, with respect to said second binding

[2055] 2855 module further embodiments of said VHH antibody domain or fragment thereof of said second

[2056] 2856 binding module are as defined in embodiments 189 to 374.

[2057] 2857

[2058] 2858 Eleventh aspect of the present disclosure (also referred to as "Embodiment 379"): According to

[2059] 2859 an eleventh aspect, the present disclosure relates to a compound comprising

[2060] 2860 - a first binding module which is a VHH antibody domain or fragment thereof, wherein

[2061] 2861 (A) said VHH antibody domain consists of the amino acid sequence of a VHH

[2062] 2862 selected from the group consisting of VHH1, VHH2, VHH3, VHH4, VHH5,

[2063] 2863 VHH6, VHH8, VHH9, VHH10 and VHH11 as shown in the Table of VHH

[2064] 2864 Sequences;

[2065] 2865 (B) said VHH antibody domain consists of a VHH sequence as defined in (A)

[2066] 2866 with modification, wherein the modification is that said sequence is

[2067] 2867 humanized;

[2068] 2868 (C) said VHH antibody domain consists of a VHH sequence as defined in (A)

[2069] 2869 with modification, wherein the modification is the replacement, addition or

[2070] 2870 deletion of up to 25 amino acids; or

[2071] 2871 (D) said VHH antibody domain consists of a VHH sequence that is at least 75%

[2072] 2872 identical to a VHH sequence referred to in (A);

[2073] 2873 - a second binding module which is a VHH antibody domain or a fragment thereof, wherein

[2074] 2874 (E) said VHH antibody domain consists of the amino acid sequence of a VHH

[2075] 2875 selected from the group consisting of VHH12, VHH13, VHH14, VHH15,

[2076] 2876 VHH16, VHH17, VHH18, VHH20, VHH21 and VHH22 as shown in the

[2077] 2877 Table of VHH Sequences; 2878 (F) said VHH antibody domain consists of a VHH sequence as defined in (E)

[2078] 2879 with modification, wherein the modification is that said sequence is

[2079] 2880 humanized;

[2080] 2881 (G) said VHH antibody domain consists of a VHH sequence as defined in (E)

[2081] 2882 with modification, wherein the modification is the replacement, addition or

[2082] 2883 deletion of up to 25 amino acids; or

[2083] 2884 (H) said VHH antibody domain consists of a VHH sequence that is at least 75%

[2084] 2885 identical to a VHH sequence referred to in (E).

[2085] 2886

[2086] 2887 To the eleventh aspect of the present disclosure and the embodiments referring thereto, the same

[2087] 2888 explanations and definitions apply accordingly as for the first to tenth aspects of the present

[2088] 2889 disclosure and the embodiments referring thereto.

[2089] 2890 For embodiment 379 and embodiments referring thereto, with respect to said first binding

[2090] 2891 module further embodiments of said VHH antibody domain or fragment thereof of said first

[2091] 2892 binding module are as defined in embodiments 1 to 188.

[2092] 2893

[2093] 2894 For embodiment 379 and embodiments referring thereto, with respect to said second binding

[2094] 2895 module further embodiments of said VHH antibody domain or fragment thereof of said second

[2095] 2896 binding module are as defined in embodiments 189 to 374.

[2096] 2897

[2097] 2898 "Compound", as used in the present disclosure, is not particularly limited and refers to a

[2098] 2899 chemical entity of any chemical class, provided that it includes the binding module(s) as defined

[2099] 2900 above. Thus, the compound can e.g. be an organic compound or a compound composed of an

[2100] 2901 organic and an inorganic part, it can be a protein composed of a single amino acid chain, a

[2101] 2902 protein composed of multiple amino acid chains that are either non-covalently or covalently

[2102] 2903 associated, or a non-covalent complex including an inorganic component. The compound can

[2103] 2904 consist of the amino acid sequence of the binding module(s) referred to in the above

[2104] 2905 embodiments alone or it can in addition include further amino acid(s) that may be covalently

[2105] 2906 or non-covalently attached, or it can be associated with inorganic components. Preferably, the

[2106] 2907 compound is a molecule.

[2107] 2908

[2108] 2909 For example, in embodiment 375, with optional modifications from embodiments 1 to 188 and

[2109] 2910 further embodiments referring to embodiment 375, (resp. in embodiment 376, with optional

[2110] 2911 modifications from embodiments 189 to 374 and further embodiments referring to embodiment 2912 376), the compound can be e.g. a monovalent, monospecific antibody molecule in which an

[2111] 2913 anti-IL-18Ra VHH according to the present disclosure (resp. an anti-IL-18RP VHH according

[2112] 2914 to the present disclosure) is linked covalently to an Fc region of an IgG. Alternatively, it could

[2113] 2915 be a bispecific molecule in which an anti-IL-18Ra VHH according to the present disclosure

[2114] 2916 (resp. an anti-IL-18RP VHH according to the present disclosure) is linked covalently to an IgGl

[2115] 2917 antibody lacking one of its "arms", such that the molecule has two binding modules, namely

[2116] 2918 the anti-IL-18Ra VHH according to the present disclosure (resp. anti-IL-18Rp VHH according

[2117] 2919 to the present disclosure) and the remaining "original" binding module of the "one-armed" IgGl

[2118] 2920 antibody. Or in another example, the compound can be a bispecific molecule comprising the

[2119] 2921 anti-IL-18Ra VHH according to the present disclosure (resp. anti-IL-18RP VHH according to

[2120] 2922 the present disclosure) and a targeting module.

[2121] 2923

[2122] 2924 For example, in embodiments 377 to 379 (with optional modifications for the first binding

[2123] 2925 module defined in embodiments 1 to 188 and optional modifications for the second binding

[2124] 2926 module defined in embodiments 189 to 374, with further optional modifications defined in the

[2125] 2927 embodiments referring to embodiments 377 to 379), the compound may e.g. be an antibody

[2126] 2928 with two binding modules (an anti-IL-18Ra VHH according to the present disclosure and an

[2127] 2929 anti-IL-18RP VHH according to the present disclosure) prepared in the SEED format, resulting

[2128] 2930 in a bispecific antibody with a structure e.g. as shown in Fig. 4A (left). The compound of

[2129] 2931 embodiment 377 to 379 may in addition comprise further molecular components, e.g. further

[2130] 2932 copies of the first and second bindings modules as exemplified in Fig. 4A (middle and right)

[2131] 2933 where molecules with two copies of the first binding module and two copies of the second

[2132] 2934 binding module in different molecular arrangements are shown. As the skilled person will

[2133] 2935 understand, many other formats of the compound are possible, provided that the resulting

[2134] 2936 compound format does not interfere with the function of the VHH antibody domain(s) or

[2135] 2937 fragment(s) thereof according to the invention (i.e. in the case of the compound of embodiment

[2136] 2938 377 to 379: the molecular format does not interfere with the binding of the first binding module

[2137] 2939 to IL18Ra and the binding of the second binding module to IL18RP, as described in the present

[2138] 2940 disclosure).

[2139] 2941

[2140] 2942 The compound of the present disclosure can be prepared by standard methods of genetic

[2141] 2943 engineering and recombinant protein technology known to the skilled person (see e.g. Green

[2142] 2944 and Sambrook, "Molecular Cloning: A Laboratory Manual", 2014; Coligan et al., "Current 2945 Protocols in Protein Science", 1997). Exemplary methods are also described in the Examples

[2143] 2946 section of the present disclosure.

[2144] 2947

[2145] 2948 In cases where the compound cannot be expressed in a single piece, individual parts can be

[2146] 2949 prepared individually and later either covalently coupled, for example by a chemical reaction

[2147] 2950 with appropriate reactive groups (e.g. linkage by maleimide chemistry) or by enzymatic linkage

[2148] 2951 (e.g. transglutaminase-catalyzed linkage). For example, the VHH antibody domain(s) or

[2149] 2952 fragment(s) thereof can be prepared by recombinant protein expression and subsequently linked

[2150] 2953 to an antibody or antibody fragment, resulting in a bispecific antibody compound as described

[2151] 2954 in the Examples section.

[2152] 2955

[2153] 2956 If the compound comprises components that are not biomolecules (such as a peptide mimetics

[2154] 2957 or a small molecule), these components may be obtained e.g. by standard methods of synthetic

[2155] 2958 organic chemistry.

[2156] 2959

[2157] 2960 Embodiment 380: The compound according to embodiment 377,

[2158] 2961 - wherein said VHH antibody domain or fragment thereof defined in (a) comprises the

[2159] 2962 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH selected

[2160] 2963 from the group consisting of VHH1, VHH2, VHH5, VHH6, VHH8, VHH9 and VHH10

[2161] 2964 as shown in the Table of CDRs and

[2162] 2965 - wherein said VHH antibody domain or fragment thereof defined in (d) comprises the

[2163] 2966 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH selected

[2164] 2967 from the group consisting of VHH12, VHH13, VHH14, VHH15, VHH17, VHH18,

[2165] 2968 VHH20, VHH21 and VHH22 as shown in the Table of CDRs, wherein said VHH

[2166] 2969 antibody domain or fragment thereof defined in (d) is selected as follows:

[2167] 2970 if the VHH antibody domain of (a) or the fragment thereof comprises the

[2168] 2971 complementarity determining regions CDR1, CDR2 and CDR3 of VHH1, then

[2169] 2972 the VHH antibody domain of (d) or the fragment thereof comprises the

[2170] 2973 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH

[2171] 2974 selected from the group consisting of VHH12, VHH13, VHH15, VHH17,

[2172] 2975 VHH20, VHH21 and VHH22 as shown in the Table of CDRs;

[2173] 2976 if the VHH antibody domain of (a) or the fragment thereof comprises the

[2174] 2977 complementarity determining regions CDR1, CDR2 and CDR3 of VHH2, then

[2175] 2978 the VHH antibody domain of (d) or the fragment thereof comprises the 2979 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH

[2176] 2980 selected from the group consisting of VHH12, VHH13, VHH15, VHH17,

[2177] 2981 VHH18, VHH20, VHH21 and VHH22 as shown in the Table of CDRs;

[2178] 2982 if the VHH antibody domain of (a) or the fragment thereof comprises the

[2179] 2983 complementarity determining regions CDR1, CDR2 and CDR3 of VHH5, then

[2180] 2984 the VHH antibody domain of (d) or the fragment thereof comprises the

[2181] 2985 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH

[2182] 2986 selected from the group consisting of VHH13 and VHH14 as shown in the Table

[2183] 2987 of CDRs;

[2184] 2988 if the VHH antibody domain of (a) or the fragment thereof comprises the

[2185] 2989 complementarity determining regions CDR1, CDR2 and CDR3 of VHH6, then

[2186] 2990 the VHH antibody domain of (d) or the fragment thereof comprises the

[2187] 2991 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH

[2188] 2992 selected from the group consisting of VHH17, VHH20, VHH21 and VHH22 as

[2189] 2993 shown in the Table of CDRs;

[2190] 2994 if the VHH antibody domain of (a) or the fragment thereof comprises the

[2191] 2995 complementarity determining regions CDR1, CDR2 and CDR3 of VHH8, then

[2192] 2996 the VHH antibody domain of (d) or the fragment thereof comprises the

[2193] 2997 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH

[2194] 2998 selected from the group consisting of VHH12, VHH13, VHH17, VHH18, VHH20

[2195] 2999 and VHH21 as shown in the Table of CDRs;

[2196] 3000 if the VHH antibody domain of (a) or the fragment thereof comprises the

[2197] 3001 complementarity determining regions CDR1, CDR2 and CDR3 of VHH9, then

[2198] 3002 the VHH antibody domain of (d) or the fragment thereof comprises the

[2199] 3003 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH

[2200] 3004 selected from the group consisting of VHH12, VHH13, VHH15, VHH17,

[2201] 3005 VHH18, VHH20, VHH21 and VHH22 as shown in the Table of CDRs;

[2202] 3006 if the VHH antibody domain of (a) or the fragment thereof comprises the

[2203] 3007 complementarity determining regions CDR1, CDR2 and CDR3 of VHHIO, then

[2204] 3008 the VHH antibody domain of (d) or the fragment thereof comprises the

[2205] 3009 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH

[2206] 3010 selected from the group consisting of VHH13, VHH20 and VHH21 as shown in

[2207] 3011 the Table of CDRs.

[2208] 3012 3013 Thus, according this embodiment, the compound of embodiment 377 (and the embodiments

[2209] 3014 referring thereto) can comprise any of the combinations VHHa and VHHP shown in Fig. 4B in

[2210] 3015 green (surrogate agonists triggering either a normalized NFKB reporter activation of more than

[2211] 3016 50% relative to IL18 or displaying potencies of less than 0.1 nM).

[2212] 3017

[2213] 3018 Embodiment 381 : The compound according to embodiment 377,

[2214] 3019 - wherein said VHH antibody domain or fragment thereof defined in (a) comprises the

[2215] 3020 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH selected

[2216] 3021 from the group consisting of VHH1, VHH2, VHH3, VHH4, VHH5, VHH6, VHH8,

[2217] 3022 VHH9 and VHH10 as shown in the Table of CDRs and

[2218] 3023 - wherein said VHH antibody domain or fragment thereof defined in (d) comprises the

[2219] 3024 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH selected

[2220] 3025 from the group consisting of VHH12, VHH13, VHH14, VHH15, VHH16, VHH17,

[2221] 3026 VHH18, VHH20, VHH21 and VHH22 as shown in the Table of CDRs, wherein said

[2222] 3027 VHH antibody domain or fragment thereof defined in (d) is selected as follows:

[2223] 3028 if the VHH antibody domain of (a) or the fragment thereof comprises the

[2224] 3029 complementarity determining regions CDR1, CDR2 and CDR3 of VHH1, then

[2225] 3030 the VHH antibody domain of (d) or the fragment thereof comprises the

[2226] 3031 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH

[2227] 3032 selected from the group consisting of VHH14, VHH16 and VHH18 as shown in

[2228] 3033 the Table of CDRs;

[2229] 3034 if the VHH antibody domain of (a) or the fragment thereof comprises the

[2230] 3035 complementarity determining regions CDR1, CDR2 and CDR3 of VHH2, then

[2231] 3036 the VHH antibody domain of (d) or the fragment thereof comprises the

[2232] 3037 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH

[2233] 3038 selected from the group consisting of VHH14 and VHH16 as shown in the Table

[2234] 3039 of CDRs;

[2235] 3040 if the VHH antibody domain of (a) or the fragment thereof comprises the

[2236] 3041 complementarity determining regions CDR1, CDR2 and CDR3 of VHH3, then

[2237] 3042 the VHH antibody domain of (d) or the fragment thereof comprises the

[2238] 3043 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH

[2239] 3044 selected from the group consisting of VHH12, VHH13, VHH14, VHH15,

[2240] 3045 VHH16, VHH17, VHH18, VHH20, VHH21 and VHH22 as shown in the Table

[2241] 3046 of CDRs; 3047 if the VHH antibody domain of (a) or the fragment thereof comprises the

[2242] 3048 complementarity determining regions CDR1, CDR2 and CDR3 of VHH4, then

[2243] 3049 the VHH antibody domain of (d) or the fragment thereof comprises the

[2244] 3050 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH

[2245] 3051 selected from the group consisting of VHH12, VHH13, VHH14, VHH15,

[2246] 3052 VHH16, VHH17, VHH18, VHH20, VHH21 and VHH22 as shown in the Table

[2247] 3053 of CDRs;

[2248] 3054 if the VHH antibody domain of (a) or the fragment thereof comprises the

[2249] 3055 complementarity determining regions CDR1, CDR2 and CDR3 of VHH5, then

[2250] 3056 the VHH antibody domain of (d) or the fragment thereof comprises the

[2251] 3057 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH

[2252] 3058 selected from the group consisting of of VHH12, VHH15, VHH16, VHH17,

[2253] 3059 VHH18, VHH20, VHH21 and VHH22 as shown in the Table of CDRs;

[2254] 3060 if the VHH antibody domain of (a) or the fragment thereof comprises the

[2255] 3061 complementarity determining regions CDR1, CDR2 and CDR3 of VHH6, then

[2256] 3062 the VHH antibody domain of (d) or the fragment thereof comprises the

[2257] 3063 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH

[2258] 3064 selected from the group consisting of VHH14, VHH16 and VHH18 as shown in

[2259] 3065 the Table of CDRs;

[2260] 3066 if the VHH antibody domain of (a) or the fragment thereof comprises the

[2261] 3067 complementarity determining regions CDR1, CDR2 and CDR3 of VHH8, then

[2262] 3068 the VHH antibody domain of (d) or the fragment thereof comprises the

[2263] 3069 complementarity determining regions CDR1, CDR2 and CDR3 of VHH16 as

[2264] 3070 shown in the Table of CDRs;

[2265] 3071 if the VHH antibody domain of (a) or the fragment thereof comprises the

[2266] 3072 complementarity determining regions CDR1, CDR2 and CDR3 of VHH9, then

[2267] 3073 the VHH antibody domain of (d) or the fragment thereof comprises the

[2268] 3074 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH

[2269] 3075 selected from the group consisting of VHH14, VHH16 and VHH18 as shown in

[2270] 3076 the Table of CDRs;

[2271] 3077 if the VHH antibody domain of (a) or the fragment thereof comprises the

[2272] 3078 complementarity determining regions CDR1, CDR2 and CDR3 of VHH10, then

[2273] 3079 the VHH antibody domain of (d) or the fragment thereof comprises the

[2274] 3080 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH 3081 selected from the group consisting of VHH14, VHH15, VHH16 and VHH18 as

[2275] 3082 shown in the Table of CDRs.

[2276] 3083

[2277] 3084 Thus, according to this embodiment, the compound of embodiment 377 (and the embodiments

[2278] 3085 referring thereto) can comprise any of the combinations VHHa and VHHP shown in Fig. 4B in

[2279] 3086 red.

[2280] 3087

[2281] 3088 Embodiment 382: The compound according to embodiment 377,

[2282] 3089 - wherein said VHH antibody domain or fragment thereof defined in (a) comprises the

[2283] 3090 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH selected

[2284] 3091 from the group consisting of VHH6 and VHH11 as shown in the Table of CDRs and

[2285] 3092 - wherein said VHH antibody domain or fragment thereof defined in (d) comprises the

[2286] 3093 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH selected

[2287] 3094 from the group consisting of VHH12, VHH13, VHH14, VHH15, VHH16, VHH17,

[2288] 3095 VHH18, VHH20, VHH21 and VHH22 as shown in the Table of CDRs, wherein said

[2289] 3096 VHH antibody domain or fragment thereof defined in (d) is selected as follows:

[2290] 3097 if the VHH antibody domain of (a) or the fragment thereof comprises the

[2291] 3098 complementarity determining regions CDR1, CDR2 and CDR3 of VHH6, then

[2292] 3099 the VHH antibody domain of (d) or the fragment thereof comprises the

[2293] 3100 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH

[2294] 3101 selected from the group consisting of VHH12 and VHH13 as shown in the Table

[2295] 3102 of CDRs;

[2296] 3103 if the VHH antibody domain of (a) or the fragment thereof comprises the

[2297] 3104 complementarity determining regions CDR1, CDR2 and CDR3 of VHH11, then

[2298] 3105 the VHH antibody domain of (d) or the fragment thereof comprises the

[2299] 3106 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH

[2300] 3107 selected from the group consisting of VHH12, VHH13, VHH14, VHH15,

[2301] 3108 VHH16, VHH17, VHH18, VHH20, VHH21 and VHH22 as shown in the Table

[2302] 3109 of CDRs.

[2303] 3110

[2304] 3111 Thus, according to this embodiment, the compound of embodiment 377 (and the embodiments

[2305] 3112 referring thereto) can comprise any of the combinations VHHa and VHHP shown in Fig. 4B in

[2306] 3113 black. 3115 Embodiment 383: The compound according to embodiment 377,

[2307] 3116 - wherein said VHH antibody domain or fragment thereof defined in (a) comprises the

[2308] 3117 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH selected

[2309] 3118 from the group consisting of VHH2, VHH6, VHH8 and VHH10 as shown in the Table

[2310] 3119 of CDRs and

[2311] 3120 - wherein said VHH antibody domain or fragment thereof defined in (d) comprises the

[2312] 3121 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH selected

[2313] 3122 from the group consisting of VHH 12, VHH 14, VHH 15, VHH 17 and VHH22 as shown

[2314] 3123 in the Table of CDRs, wherein said VHH antibody domain or fragment thereof defined

[2315] 3124 in (d) is selected as follows:

[2316] 3125 if the VHH antibody domain of (a) or the fragment thereof comprises the

[2317] 3126 complementarity determining regions CDR1, CDR2 and CDR3 of VHH2, then

[2318] 3127 the VHH antibody domain of (d) or the fragment thereof comprises the

[2319] 3128 complementarity determining regions CDR1, CDR2 and CDR3 of VHH22 as

[2320] 3129 shown in the Table of CDRs;

[2321] 3130 if the VHH antibody domain of (a) or the fragment thereof comprises the

[2322] 3131 complementarity determining regions CDR1, CDR2 and CDR3 of VHH6, then

[2323] 3132 the VHH antibody domain of (d) or the fragment thereof comprises the

[2324] 3133 complementarity determining regions CDR1, CDR2 and CDR3 of VHH15 as

[2325] 3134 shown in the Table of CDRs;

[2326] 3135 if the VHH antibody domain of (a) or the fragment thereof comprises the

[2327] 3136 complementarity determining regions CDR1, CDR2 and CDR3 of VHH8, then

[2328] 3137 the VHH antibody domain of (d) or the fragment thereof comprises the

[2329] 3138 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH

[2330] 3139 selected from the group consisting of VHH14, VHH15 and VHH22 as shown in

[2331] 3140 the Table of CDRs;

[2332] 3141 if the VHH antibody domain of (a) or the fragment thereof comprises the

[2333] 3142 complementarity determining regions CDR1, CDR2 and CDR3 of VHH10, then

[2334] 3143 the VHH antibody domain of (d) or the fragment thereof comprises the

[2335] 3144 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH

[2336] 3145 selected from the group consisting of VHH12, VHH17 and VHH22 as shown in

[2337] 3146 the Table of CDRs.

[2338] 3147 3148 Thus, according to this embodiment, the compound of embodiment 377 (and the embodiments

[2339] 3149 referring thereto) can comprise any of the combinations VHHa and VHHP shown in Fig. 4B in

[2340] 3150 orange (active surrogate agonists eliciting less than 50% of NFKB reporter activation

[2341] 3151 normalized to (rh) IL-18 at 1 nM or EC50 higher than 0.1 nM).

[2342] 3152

[2343] 3153 Embodiment 384: The compound according to embodiment 377, wherein said VHH antibody

[2344] 3154 domain or fragment thereof defined in (a) comprises the complementarity determining regions

[2345] 3155 CDR1, CDR2 and CDR3 of one VHH selected from the group consisting of VHH2 and VHH8

[2346] 3156 as shown in the Table of CDRs and wherein said VHH antibody domain or fragment thereof

[2347] 3157 defined in (d) comprises the complementarity determining regions CDR1, CDR2 and CDR3 of

[2348] 3158 one VHH selected from the group consisting of VHH15 and VHH17 as shown in the Table of

[2349] 3159 CDRs.

[2350] 3160

[2351] 3161 Embodiment 385: The compound according to embodiment 377,

[2352] 3162 - wherein said VHH antibody domain or fragment thereof defined in (a) comprises the

[2353] 3163 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH selected

[2354] 3164 from the group consisting of VHH2 and VHH8 as shown in the Table of CDRs and

[2355] 3165 - wherein said VHH antibody domain or fragment thereof defined in (d) comprises the

[2356] 3166 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH selected

[2357] 3167 from the group consisting of VHH15 and VHH17 as shown in the Table of CDRs,

[2358] 3168 wherein said VHH antibody domain or fragment thereof defined in (d) is selected as

[2359] 3169 follows:

[2360] 3170 if the VHH antibody domain of (a) or the fragment thereof comprises the

[2361] 3171 complementarity determining regions CDR1, CDR2 and CDR3 of VHH2, then

[2362] 3172 the VHH antibody domain of (d) or the fragment thereof comprises the

[2363] 3173 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH

[2364] 3174 selected from the group consisting of VHH15 and VHH17 as shown in the Table

[2365] 3175 of CDRs;

[2366] 3176 if the VHH antibody domain of (a) or the fragment thereof comprises the

[2367] 3177 complementarity determining regions CDR1, CDR2 and CDR3 of VHH8, then

[2368] 3178 the VHH antibody domain of (d) or the fragment thereof comprises the

[2369] 3179 complementarity determining regions CDR1, CDR2 and CDR3 of VHH17 as

[2370] 3180 shown in the Table of CDRs.

[2371] 3181 3182 Embodiment 386: The compound according to embodiment 377, wherein said VHH antibody

[2372] 3183 domain or fragment thereof defined in (a) comprises the complementarity determining regions

[2373] 3184 CDR1, CDR2 and CDR3 of VHH2 as shown in the Table of CDRs and wherein said VHH

[2374] 3185 antibody domain or fragment thereof defined in (d) comprises the complementarity determining

[2375] 3186 regions CDR1, CDR2 and CDR3 of VHH15 as shown in the Table of CDRs.

[2376] 3187

[2377] 3188 Embodiment 387: The compound according to embodiment 377,

[2378] 3189 - wherein said VHH antibody domain or fragment thereof defined in (a) comprises the

[2379] 3190 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH selected

[2380] 3191 from the group consisting of VHH1, VHH2, VHH5, VHH6, VHH8 and VHH 10 as

[2381] 3192 shown in the Table of CDRs and

[2382] 3193 - wherein said VHH antibody domain or fragment thereof defined in (d) comprises the

[2383] 3194 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH selected

[2384] 3195 from the group consisting of VHH13, VHH16 and VHH22 as shown in the Table of

[2385] 3196 CDRs, wherein said VHH antibody domain or fragment thereof defined in (d) is selected

[2386] 3197 as follows:

[2387] 3198 if the VHH antibody domain of (a) or the fragment thereof comprises the

[2388] 3199 complementarity determining regions CDR1, CDR2 and CDR3 of VHH1, then

[2389] 3200 the VHH antibody domain of (d) or the fragment thereof comprises the

[2390] 3201 complementarity determining regions CDR1, CDR2 and CDR3 of VHH16 as

[2391] 3202 shown in the Table of CDRs;

[2392] 3203 if the VHH antibody domain of (a) or the fragment thereof comprises the

[2393] 3204 complementarity determining regions CDR1, CDR2 and CDR3 of VHH2, then

[2394] 3205 the VHH antibody domain of (d) or the fragment thereof comprises the

[2395] 3206 complementarity determining regions CDR1, CDR2 and CDR3 of VHH22 as

[2396] 3207 shown in the Table of CDRs;

[2397] 3208 if the VHH antibody domain of (a) or the fragment thereof comprises the

[2398] 3209 complementarity determining regions CDR1, CDR2 and CDR3 of VHH5, then

[2399] 3210 the VHH antibody domain of (d) or the fragment thereof comprises the

[2400] 3211 complementarity determining regions CDR1, CDR2 and CDR3 of VHH13 as

[2401] 3212 shown in the Table of CDRs;

[2402] 3213 if the VHH antibody domain of (a) or the fragment thereof comprises the

[2403] 3214 complementarity determining regions CDR1, CDR2 and CDR3 of VHH6, then

[2404] 3215 the VHH antibody domain of (d) or the fragment thereof comprises the 3216 complementarity determining regions CDR1, CDR2 and CDR3 of VHH22 as

[2405] 3217 shown in the Table of CDRs;

[2406] 3218 if the VHH antibody domain of (a) or the fragment thereof comprises the

[2407] 3219 complementarity determining regions CDR1, CDR2 and CDR3 of VHH8, then

[2408] 3220 the VHH antibody domain of (d) or the fragment thereof comprises the

[2409] 3221 complementarity determining regions CDR1, CDR2 and CDR3 of VHH22 as

[2410] 3222 shown in the Table of CDRs;

[2411] 3223 if the VHH antibody domain of (a) or the fragment thereof comprises the

[2412] 3224 complementarity determining regions CDR1, CDR2 and CDR3 of VHH10, then

[2413] 3225 the VHH antibody domain of (d) or the fragment thereof comprises the

[2414] 3226 complementarity determining regions CDR1, CDR2 and CDR3 of VHH22 as

[2415] 3227 shown in the Table of CDRs.

[2416] 3228

[2417] 3229 Embodiment 388: The compound according to embodiment 377,

[2418] 3230 - wherein said VHH antibody domain or fragment thereof defined in (a) comprises the

[2419] 3231 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH selected

[2420] 3232 from the group consisting of VHH2, VHH6, VHH8 and VHH10 as shown in the Table

[2421] 3233 of CDRs and

[2422] 3234 - wherein said VHH antibody domain or fragment thereof defined in (d) comprises the

[2423] 3235 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH selected

[2424] 3236 from the group consisting of VHH12 and VHH15 as shown in the Table of CDRs, wherein

[2425] 3237 said VHH antibody domain or fragment thereof defined in (d) is selected as follows:

[2426] 3238 if the VHH antibody domain of (a) or the fragment thereof comprises the

[2427] 3239 complementarity determining regions CDR1, CDR2 and CDR3 of VHH2, then the

[2428] 3240 VHH antibody domain of (d) or the fragment thereof comprises the

[2429] 3241 complementarity determining regions CDR1, CDR2 and CDR3 of VHH12 as

[2430] 3242 shown in the Table of CDRs;

[2431] 3243 if the VHH antibody domain of (a) or the fragment thereof comprises the

[2432] 3244 complementarity determining regions CDR1, CDR2 and CDR3 of VHH6, then the

[2433] 3245 VHH antibody domain of (d) or the fragment thereof comprises the

[2434] 3246 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH

[2435] 3247 selected from the group consisting of VHH12 and VHH15 as shown in the Table of

[2436] 3248 CDRs; 3249 if the VHH antibody domain of (a) or the fragment thereof comprises the

[2437] 3250 complementarity determining regions CDR1, CDR2 and CDR3 of VHH8, then the

[2438] 3251 VHH antibody domain of (d) or the fragment thereof comprises the

[2439] 3252 complementarity determining regions CDR1, CDR2 and CDR3 of one VHH

[2440] 3253 selected from the group consisting of VHH13 and VHH18 as shown in the Table of

[2441] 3254 CDRs;

[2442] 3255 if the VHH antibody domain of (a) or the fragment thereof comprises the

[2443] 3256 complementarity determining regions CDR1, CDR2 and CDR3 of VHH10, then

[2444] 3257 the VHH antibody domain of (d) or the fragment thereof comprises the

[2445] 3258 complementarity determining regions CDR1, CDR2 and CDR3 of VHH12 as

[2446] 3259 shown in the Table of CDRs.

[2447] 3260

[2448] 3261 Embodiment 389: The compound according to any one of embodiments 377 or 380 to 388,

[2449] 3262 wherein in the first binding module said VHH antibody domain or fragment thereof comprises

[2450] 3263 complementarity determining regions according to (a) or (b).

[2451] 3264

[2452] 3265 If the present disclosure states that "in the first binding module said VHH antibody domain or

[2453] 3266 fragment thereof comprises complementarity determining regions according to (a) or (b)", then

[2454] 3267 this is meant to designate that in this embodiment only (a) and (b) are considered as options for

[2455] 3268 the VHH antibody domain (or fragment thereof) of the first binding module, whereas option (c)

[2456] 3269 is excluded. If the present disclosure states that "in the first binding module said VHH antibody

[2457] 3270 domain or fragment thereof comprises complementarity determining regions according to (a)

[2458] 3271 or (c)", then this is meant to designate that in this embodiment only (a) and (c) are considered

[2459] 3272 as options for the VHH antibody domain (or fragment thereof) of the first binding module,

[2460] 3273 whereas option (b) is excluded. The alternative wording below is to be understood accordingly.

[2461] 3274 If the present disclosure states that "in the first binding module said VHH antibody domain or

[2462] 3275 fragment thereof comprises complementarity determining regions according to (a)", then this

[2463] 3276 is meant to designate that in this embodiment only (a) is considered as option for the VHH

[2464] 3277 antibody domain (or fragment thereof) of the first binding module, whereas options (b) and (c)

[2465] 3278 are excluded. Further variations of this wording below are to be understood accordingly.

[2466] 3279

[2467] 3280 Embodiment 390: The compound according to any one of embodiments 377 or 380 to 388,

[2468] 3281 wherein in the first binding module said VHH antibody domain or fragment thereof comprises

[2469] 3282 complementarity determining regions according to (a) or (c). 3283

[2470] 3284 Embodiment 391 : The compound according to any one of embodiments 377 or 380 to 388,

[2471] 3285 wherein in the first binding module said VHH antibody domain or fragment thereof comprises

[2472] 3286 complementarity determining regions according to (a).

[2473] 3287

[2474] 3288 Embodiment 392: The compound according to any one of embodiments 377 or 380 to 388,

[2475] 3289 wherein in the first binding module said VHH antibody domain or fragment thereof comprises

[2476] 3290 complementarity determining regions according to (b).

[2477] 3291

[2478] 3292 Embodiment 393: The compound according to any one of embodiments 377 or 380 to 392,

[2479] 3293 wherein in the second binding module said VHH antibody domain or fragment thereof

[2480] 3294 comprises complementarity determining regions according to (d) or (e).

[2481] 3295

[2482] 3296 Embodiment 394: The compound according to any one of embodiments 377 or 380 to 392,

[2483] 3297 wherein in the second binding module said VHH antibody domain or fragment thereof

[2484] 3298 comprises complementarity determining regions according to (d) or (f).

[2485] 3299

[2486] 3300 Embodiment 395: The compound according to any one of embodiments 377 or 380 to 392,

[2487] 3301 wherein in the second binding module said VHH antibody domain or fragment thereof

[2488] 3302 comprises complementarity determining regions according to (d).

[2489] 3303

[2490] 3304 Embodiment 396: The compound according to any one of embodiments 377 or 380 to 392,

[2491] 3305 wherein in the second binding module said VHH antibody domain or fragment thereof

[2492] 3306 comprises complementarity determining regions according to (e).

[2493] 3307

[2494] 3308 Embodiment 397: The compound according to any one of embodiments 378 or 379,

[2495] 3309 - wherein said VHH antibody domain of (A) comprises the amino acid sequence of one

[2496] 3310 VHH selected from the group consisting of VHH1, VHH2, VHH5, VHH6, VHH8,

[2497] 3311 VHH9 and VHH10 shown in the Table of VHH Sequences and

[2498] 3312 - wherein said VHH antibody domain of (E) comprises the amino acid sequence of one

[2499] 3313 VHH selected from the group consisting of VHH12, VHH13, VHH14, VHH15,

[2500] 3314 VHH17, VHH18, VHH20, VHH21 and VHH22 shown in the Table of VHH Sequences,

[2501] 3315 wherein said VHH antibody domain of (E) is selected as follows: 3316 if the VHH antibody domain of (A) comprises the amino acid sequence of VHH1,

[2502] 3317 then the VHH antibody domain of (E) comprises the amino acid sequence of one

[2503] 3318 VHH selected from the group consisting of VHH12, VHH13, VHH15, VHH17,

[2504] 3319 VHH20, VHH21 and VHH22 shown in the Table of VHH Sequences;

[2505] 3320 if the VHH antibody domain of (A) comprises the amino acid sequence of VHH2,

[2506] 3321 then the VHH antibody domain of (E) comprises the amino acid sequence of one

[2507] 3322 VHH selected from the group consisting of VHH12, VHH13, VHH15, VHH17,

[2508] 3323 VHH18, VHH20, VHH21 and VHH22 shown in the Table of VHH Sequences;

[2509] 3324 if the VHH antibody domain of (A) comprises the amino acid sequence of VHH5,

[2510] 3325 then the VHH antibody domain of (E) comprises the amino acid sequence of one

[2511] 3326 VHH selected from the group consisting of VHH13 and VHH14 shown in the

[2512] 3327 Table of VHH Sequences;

[2513] 3328 if the VHH antibody domain of (A) comprises the amino acid sequence of VHH6,

[2514] 3329 then the VHH antibody domain of (E) comprises the amino acid sequence of one

[2515] 3330 VHH selected from the group consisting of VHH17, VHH20, VHH21 and VHH22

[2516] 3331 shown in the Table of VHH Sequences;

[2517] 3332 if the VHH antibody domain of (A) comprises the amino acid sequence of VHH8,

[2518] 3333 then the VHH antibody domain of (E) comprises the amino acid sequence of one

[2519] 3334 VHH selected from the group consisting of VHH12, VHH13, VHH17, VHH18,

[2520] 3335 VHH20 and VHH21 shown in the Table of VHH Sequences;

[2521] 3336 if the VHH antibody domain of (A) comprises the amino acid sequence of VHH9,

[2522] 3337 then the VHH antibody domain of (E) comprises the amino acid sequence of one

[2523] 3338 VHH selected from the group consisting of VHH12, VHH13, VHH15, VHH17,

[2524] 3339 VHH18, VHH20, VHH21 and VHH22 shown in the Table of VHH Sequences;

[2525] 3340 if the VHH antibody domain of (A) comprises the amino acid sequence of VHH10,

[2526] 3341 then the VHH antibody domain of (E) comprises the amino acid sequence of one

[2527] 3342 VHH selected from the group consisting of VHH13, VHH20 and VHH21 shown

[2528] 3343 in the Table of VHH Sequences.

[2529] 3344

[2530] 3345 Thus, according to this embodiment, the compound of embodiments 378 to 379 (and the

[2531] 3346 embodiments referring thereto) can comprise any of the combinations VHHa and VHHP shown

[2532] 3347 in Fig. 4B in green (surrogate agonists triggering either a normalized NFKB reporter activation

[2533] 3348 of more than 50% relative to IL18 or displaying potencies of less than 0.1 nM).

[2534] 3349 3350 Embodiment 398: The compound according to any one of embodiments 378 or 379,

[2535] 3351 - wherein said VHH antibody domain of (A) comprises the amino acid sequence of one

[2536] 3352 VHH selected from the group consisting of VHH1, VHH2, VHH3, VHH4, VHH5,

[2537] 3353 VHH6, VHH8, VHH9 and VHH10 shown in the Table of VHH Sequences and

[2538] 3354 - wherein said VHH antibody domain of (E) comprises the amino acid sequence of one

[2539] 3355 VHH selected from the group consisting of VHH12, VHH13, VHH14, VHH15,

[2540] 3356 VHH16, VHH17, VHH18, VHH20, VHH21 and VHH22 shown in the Table of VHH

[2541] 3357 Sequences, wherein said VHH antibody domain of (E) is selected as follows:

[2542] 3358 if the VHH antibody domain of (A) comprises the amino acid sequence of VHH1,

[2543] 3359 then the VHH antibody domain of (E) comprises the amino acid sequence of one

[2544] 3360 VHH selected from the group consisting of VHH14, VHH16 and VHH18 shown

[2545] 3361 in the Table of VHH Sequences;

[2546] 3362 if the VHH antibody domain of (A) comprises the amino acid sequence of VHH2,

[2547] 3363 then the VHH antibody domain of (E) comprises the amino acid sequence of one

[2548] 3364 VHH selected from the group consisting of VHH14 and VHH16 shown in the

[2549] 3365 Table of VHH Sequences;

[2550] 3366 if the VHH antibody domain of (A) comprises the amino acid sequence of VHH3,

[2551] 3367 then the VHH antibody domain of (E) comprises the amino acid sequence of one

[2552] 3368 VHH selected from the group consisting of VHH12, VHH13, VHH14, VHH15,

[2553] 3369 VHH16, VHH17, VHH18, VHH20, VHH21 and VHH22 shown in the Table of

[2554] 3370 VHH Sequences;

[2555] 3371 if the VHH antibody domain of (A) comprises the amino acid sequence of VHH4,

[2556] 3372 then the VHH antibody domain of (E) comprises the amino acid sequence of one

[2557] 3373 VHH selected from the group consisting of VHH12, VHH13, VHH14, VHH15,

[2558] 3374 VHH16, VHH17, VHH18, VHH20, VHH21 and VHH22 shown in the Table of

[2559] 3375 VHH Sequences;

[2560] 3376 if the VHH antibody domain of (A) comprises the amino acid sequence of VHH5,

[2561] 3377 then the VHH antibody domain of (E) comprises the amino acid sequence of one

[2562] 3378 VHH selected from the group consisting of VHH12, VHH15, VHH16, VHH17,

[2563] 3379 VHH18, VHH20, VHH21 and VHH22 shown in the Table of VHH Sequences;

[2564] 3380 if the VHH antibody domain of (A) comprises the amino acid sequence of VHH6,

[2565] 3381 then the VHH antibody domain of (E) comprises the amino acid sequence of one

[2566] 3382 VHH selected from the group consisting of VHH14, VHH16 and VHH18 shown

[2567] 3383 in the Table of VHH Sequences; 3384 if the VHH antibody domain of (A) comprises the amino acid sequence of VHH8,

[2568] 3385 then the VHH antibody domain of (E) comprises the amino acid sequence of

[2569] 3386 VHH16 shown in the Table of VHH Sequences;

[2570] 3387 if the VHH antibody domain of (A) comprises the amino acid sequence of VHH9,

[2571] 3388 then the VHH antibody domain of (E) comprises the amino acid sequence of one

[2572] 3389 VHH selected from the group consisting of VHH14, VHH16 and VHH18 shown

[2573] 3390 in the Table of VHH Sequences;

[2574] 3391 if the VHH antibody domain of (A) comprises the amino acid sequence of VHH10,

[2575] 3392 then the VHH antibody domain of (E) comprises the amino acid sequence of one

[2576] 3393 VHH selected from the group consisting of VHH14, VHH15, VHH16 and VHH18

[2577] 3394 shown in the Table of VHH Sequences.

[2578] 3395

[2579] 3396 Thus, according to this embodiment, the compound of embodiments 378 to 379 (and the

[2580] 3397 embodiments referring thereto) can comprise any of the combinations VHHa and VHHP shown

[2581] 3398 in Fig. 4B in red.

[2582] 3399

[2583] 3400 Embodiment 399: The compound according to any one of embodiments 378 or 379,

[2584] 3401 - wherein said VHH antibody domain of (A) comprises the amino acid sequence of one

[2585] 3402 VHH selected from the group consisting of VHH6 and VHH11 shown in the Table of

[2586] 3403 VHH Sequences and

[2587] 3404 - wherein said VHH antibody domain of (E) comprises the amino acid sequence of one

[2588] 3405 VHH selected from the group consisting of VHH12, VHH13, VHH14, VHH15,

[2589] 3406 VHH16, VHH17, VHH18, VHH20, VHH21 and VHH22 shown in the Table of VHH

[2590] 3407 Sequences, wherein said VHH antibody domain of (E) is selected as follows:

[2591] 3408 if the VHH antibody domain of (A) comprises the amino acid sequence of VHH6,

[2592] 3409 then the VHH antibody domain of (E) comprises the amino acid sequence of one

[2593] 3410 VHH selected from the group consisting of VHH12 and VHH13 shown in the

[2594] 3411 Table of VHH Sequences;

[2595] 3412 if the VHH antibody domain of (A) comprises the amino acid sequence of VHH11,

[2596] 3413 then the VHH antibody domain of (E) comprises the amino acid sequence of one

[2597] 3414 VHH selected from the group consisting of VHH12, VHH13, VHH14, VHH15,

[2598] 3415 VHH16, VHH17, VHH18, VHH20, VHH21 and VHH22 shown in the Table of

[2599] 3416 VHH Sequences.

[2600] 3417 3418 Thus, according to this embodiment, the compound of embodiments 378 to 379 (and the

[2601] 3419 embodiments referring thereto) can comprise any of the combinations VHHa and VHHP shown

[2602] 3420 in Fig. 4B in black.

[2603] 3421

[2604] 3422 Embodiment 400: The compound according to any one of embodiments 378 or 379,

[2605] 3423 - wherein said VHH antibody domain of (A) comprises the amino acid sequence of one

[2606] 3424 VHH selected from the group consisting of VHH2, VHH6, VHH8 and VHH10 shown

[2607] 3425 in the Table of VHH Sequences and

[2608] 3426 - wherein said VHH antibody domain of (E) comprises the amino acid sequence of one

[2609] 3427 VHH selected from the group consisting of VHH 12, VHH14, VHH15, VHH17 and

[2610] 3428 VHH22 shown in the Table of VHH Sequences, wherein said VHH antibody domain of

[2611] 3429 (E) is selected as follows:

[2612] 3430 if the VHH antibody domain of (A) comprises the amino acid sequence of VHH2,

[2613] 3431 then the VHH antibody domain of (E) comprises the amino acid sequence of

[2614] 3432 VHH22 shown in the Table of VHH Sequences;

[2615] 3433 if the VHH antibody domain of (A) comprises the amino acid sequence of VHH6,

[2616] 3434 then the VHH antibody domain of (E) comprises the amino acid sequence of

[2617] 3435 VHH15 shown in the Table of VHH Sequences;

[2618] 3436 if the VHH antibody domain of (A) comprises the amino acid sequence of VHH8,

[2619] 3437 then the VHH antibody domain of (E) comprises the amino acid sequence of one

[2620] 3438 VHH selected from the group consisting of VHH14, VHH15 and VHH22 shown

[2621] 3439 in the Table of VHH Sequences;

[2622] 3440 if the VHH antibody domain of (A) comprises the amino acid sequence of VHH10,

[2623] 3441 then the VHH antibody domain of (E) comprises the amino acid sequence of one

[2624] 3442 VHH selected from the group consisting of VHH12, VHH17 and VHH22 shown

[2625] 3443 in the Table of VHH Sequences.

[2626] 3444

[2627] 3445 Thus, according to this embodiment, the compound of embodiments 378 to 379 (and the

[2628] 3446 embodiments referring thereto) can comprise any of the combinations VHHa and VHHP shown

[2629] 3447 in Fig. 4B in orange (active surrogate agonists eliciting less than 50% of NFKB reporter

[2630] 3448 activation normalized to (rh) IL-18 at 1 nM or EC50 higher than 0.1 nM).

[2631] 3449

[2632] 3450 Embodiment 401 : The compound according to any one of embodiments 378 or 379, wherein

[2633] 3451 said VHH antibody domain of (A) comprises the amino acid sequence of one VHH selected 3452 from the group consisting of VHH2 and VHH8 shown in the Table of VHH Sequences and

[2634] 3453 wherein said VHH antibody domain of (E) comprises the amino acid sequence of one VHH

[2635] 3454 selected from the group consisting of VHH15 and VHH17 shown in the Table of VHH

[2636] 3455 Sequences.

[2637] 3456

[2638] 3457 Embodiment 402: The compound according to any one of embodiments 378 or 379,

[2639] 3458 - wherein said VHH antibody domain of (A) comprises the amino acid sequence of one

[2640] 3459 VHH selected from the group consisting of VHH2 and VHH8 shown in the Table of

[2641] 3460 VHH Sequences and

[2642] 3461 - wherein said VHH antibody domain of (E) comprises the amino acid sequence of one

[2643] 3462 VHH selected from the group consisting of VHH15 and VHH17 shown in the Table of

[2644] 3463 VHH Sequences, wherein said VHH antibody domain of (E) is selected as follows:

[2645] 3464 if the VHH antibody domain of (A) comprises the amino acid sequence of VHH2,

[2646] 3465 then the VHH antibody domain of (E) comprises the amino acid sequence of one

[2647] 3466 VHH selected from the group consisting of VHH15 and VHH17 shown in the

[2648] 3467 Table of VHH Sequences;

[2649] 3468 if the VHH antibody domain of (A) comprises the amino acid sequence of VHH8,

[2650] 3469 then the VHH antibody domain of (E) comprises the amino acid sequence of

[2651] 3470 VHH17 shown in the Table of VHH Sequences.

[2652] 3471

[2653] 3472 Embodiment 403: The compound according to any one of embodiments 378 or 379, wherein

[2654] 3473 said VHH antibody domain of (A) comprises the amino acid sequence of VHH2 shown in the

[2655] 3474 Table of VHH Sequences and wherein said VHH antibody domain of (E) comprises the amino

[2656] 3475 acid sequence of VHH15 shown in the Table of VHH Sequences.

[2657] 3476

[2658] 3477 Embodiment 404: The compound according to any one of embodiments 378 or 379,

[2659] 3478 - wherein said VHH antibody domain of (A) comprises the amino acid sequence of one

[2660] 3479 VHH selected from the group consisting of VHH1, VHH2, VHH5, VHH6, VHH8 and

[2661] 3480 VHH10 shown in the Table of VHH Sequences and

[2662] 3481 - wherein said VHH antibody domain of (E) comprises the amino acid sequence of one

[2663] 3482 VHH selected from the group consisting of VHH13, VHH16 and VHH22 shown in the

[2664] 3483 Table of VHH Sequences, wherein said VHH antibody domain of (E) is selected as

[2665] 3484 follows: 3485 if the VHH antibody domain of (A) comprises the amino acid sequence of VHH1,

[2666] 3486 then the VHH antibody domain of (E) comprises the amino acid sequence of

[2667] 3487 VHH16 shown in the Table of VHH Sequences;

[2668] 3488 if the VHH antibody domain of (A) comprises the amino acid sequence of VHH2,

[2669] 3489 then the VHH antibody domain of (E) comprises the amino acid sequence of

[2670] 3490 VHH22 shown in the Table of VHH Sequences;

[2671] 3491 if the VHH antibody domain of (A) comprises the amino acid sequence of VHH5,

[2672] 3492 then the VHH antibody domain of (E) comprises the amino acid sequence of

[2673] 3493 VHH13 shown in the Table of VHH Sequences;

[2674] 3494 if the VHH antibody domain of (A) comprises the amino acid sequence of VHH6,

[2675] 3495 then the VHH antibody domain of (E) comprises the amino acid sequence of

[2676] 3496 VHH22 shown in the Table of VHH Sequences;

[2677] 3497 if the VHH antibody domain of (A) comprises the amino acid sequence of VHH8,

[2678] 3498 then the VHH antibody domain of (E) comprises the amino acid sequence of

[2679] 3499 VHH22 shown in the Table of VHH Sequences;

[2680] 3500 if the VHH antibody domain of (A) comprises the amino acid sequence of VHH10,

[2681] 3501 then the VHH antibody domain of (E) comprises the amino acid sequence of

[2682] 3502 VHH22 shown in the Table of VHH Sequences.

[2683] 3503

[2684] 3504 Embodiment 405: The compound according to any one of embodiments 378 or 379,

[2685] 3505 - wherein said VHH antibody domain of (A) comprises the amino acid sequence of one

[2686] 3506 VHH selected from the group consisting of VHH2, VHH6, VHH8 and VHH10 shown

[2687] 3507 in the Table of VHH Sequences and

[2688] 3508 - wherein said VHH antibody domain of (E) comprises the amino acid sequence of one

[2689] 3509 VHH selected from the group consisting of VHH12, VHH13, VHH15 and VHH18

[2690] 3510 shown in the Table of VHH Sequences, wherein said VHH antibody domain of (E) is

[2691] 3511 selected as follows:

[2692] 3512 if the VHH antibody domain of (A) comprises the amino acid sequence of VHH2,

[2693] 3513 then the VHH antibody domain of (E) comprises the amino acid sequence of

[2694] 3514 VHH12 shown in the Table of VHH Sequences;

[2695] 3515 if the VHH antibody domain of (A) comprises the amino acid sequence of VHH6,

[2696] 3516 then the VHH antibody domain of (E) comprises the amino acid sequence of one

[2697] 3517 VHH selected from the group consisting of VHH15 and VHH12 shown in the

[2698] 3518 Table of VHH Sequences; 3519 if the VHH antibody domain of (A) comprises the amino acid sequence of VHH8,

[2699] 3520 then the VHH antibody domain of (E) comprises the amino acid sequence of one

[2700] 3521 VHH selected from the group consisting of VHH13 and VHH18 shown in the

[2701] 3522 Table of VHH Sequences;

[2702] 3523 if the VHH antibody domain of (A) comprises the amino acid sequence of VHH10,

[2703] 3524 then the VHH antibody domain of (E) comprises the amino acid sequence of

[2704] 3525 VHH12 shown in the Table of VHH Sequences.

[2705] 3526

[2706] 3527 Embodiment 406: The compound according to any one of embodiments 378 to 379 or 397 to

[2707] 3528 405, wherein in the first binding module said VHH antibody domain or fragment thereof

[2708] 3529 comprises a VHH antibody domain or fragment thereof according to (A), (B) or (C).

[2709] 3530

[2710] 3531 If the present disclosure states that " in the first binding module said VHH antibody domain or

[2711] 3532 fragment thereof comprises a VHH antibody domain or fragment thereof according to (A), (B)

[2712] 3533 or (C)", then this is meant to designate that in this embodiment only (A), (B) and (C) are

[2713] 3534 considered as options for the VHH antibody domain (or fragment thereof) of the first binding

[2714] 3535 module, whereas the option (D) is excluded. If the present disclosure states that " in the first

[2715] 3536 binding module said VHH antibody domain or fragment thereof comprises a VHH antibody

[2716] 3537 domain or fragment thereof according to (A), (C) or (D)", then this is meant to designate that

[2717] 3538 in this embodiment only (A), (C) and (D) are considered as options for the VHH antibody

[2718] 3539 domain (or fragment thereof) of the first binding module, whereas the option (B) is excluded.

[2719] 3540 If the present disclosure states that " in the first binding module said VHH antibody domain or

[2720] 3541 fragment thereof comprises a VHH antibody domain or fragment thereof according to (A)",

[2721] 3542 then this is meant to designate that in this embodiment only (A) is considered as option for the

[2722] 3543 VHH antibody domain (or fragment thereof) of the first binding module, whereas the options

[2723] 3544 (B), (C) and (D) are excluded. Further variations of this wording below are to be understood

[2724] 3545 accordingly.

[2725] 3546

[2726] 3547 Embodiment 407: The compound according to any one of embodiments 378 to 379 or 397 to

[2727] 3548 405, wherein in the first binding module said VHH antibody domain or fragment thereof

[2728] 3549 comprises a VHH antibody domain or fragment thereof according to (A), (B) or (D).

[2729] 3550 3551 Embodiment 408: The compound according to any one of embodiments 378 to 379 or 397 to

[2730] 3552 405, wherein in the first binding module said VHH antibody domain or fragment thereof

[2731] 3553 comprises a VHH antibody domain or fragment thereof according to (A), (C) or (D).

[2732] 3554

[2733] 3555 Embodiment 409: The compound according to any one of embodiments 378 to 379 or 397 to

[2734] 3556 405, wherein in the first binding module said VHH antibody domain or fragment thereof

[2735] 3557 comprises a VHH antibody domain or fragment thereof according to (A) or (B).

[2736] 3558

[2737] 3559 Embodiment 410: The compound according to any one of embodiments 378 to 379 or 397 to

[2738] 3560 405, wherein in the first binding module said VHH antibody domain or fragment thereof

[2739] 3561 comprises a VHH antibody domain or fragment thereof according to (A) or (C).

[2740] 3562

[2741] 3563 Embodiment 411 : The compound according to any one of embodiments 378 to 379 or 397 to

[2742] 3564 405, wherein in the first binding module said VHH antibody domain or fragment thereof

[2743] 3565 comprises a VHH antibody domain or fragment thereof according to (A) or (D).

[2744] 3566

[2745] 3567 Embodiment 412: The compound according to any one of embodiments 378 to 379 or 397 to

[2746] 3568 405, wherein in the first binding module said VHH antibody domain or fragment thereof

[2747] 3569 comprises a VHH antibody domain or fragment thereof according to (A).

[2748] 3570

[2749] 3571 Embodiment 413: The compound according to any one of embodiments 378 to 379 or 397 to

[2750] 3572 405, wherein in the first binding module said VHH antibody domain or fragment thereof

[2751] 3573 comprises a VHH antibody domain or fragment thereof according to (B).

[2752] 3574

[2753] 3575 Embodiment 414: The compound according to any one of embodiments 378 to 379 or 397 to

[2754] 3576 413, wherein in the second binding module said VHH antibody domain or fragment thereof

[2755] 3577 comprises a VHH antibody domain or fragment thereof according to (E), (F) or (G).

[2756] 3578

[2757] 3579 Embodiment 415: The compound according to any one of embodiments 378 to 379 or 397 to

[2758] 3580 413, wherein in the second binding module said VHH antibody domain or fragment thereof

[2759] 3581 comprises a VHH antibody domain or fragment thereof according to (E), (F) or (H).

[2760] 3582 3583 Embodiment 416: The compound according to any one of embodiments 378 to 379 or 397 to

[2761] 3584 413, wherein in the second binding module said VHH antibody domain or fragment thereof

[2762] 3585 comprises a VHH antibody domain or fragment thereof according to (E), (G) or (H).

[2763] 3586

[2764] 3587 Embodiment 417: The compound according to any one of embodiments 378 to 379 or 397 to

[2765] 3588 413, wherein in the second binding module said VHH antibody domain or fragment thereof

[2766] 3589 comprises a VHH antibody domain or fragment thereof according to (E) or (F).

[2767] 3590

[2768] 3591 Embodiment 418: The compound according to any one of embodiments 378 to 379 or 397 to

[2769] 3592 413, wherein in the second binding module said VHH antibody domain or fragment thereof

[2770] 3593 comprises a VHH antibody domain or fragment thereof according to (E) or (G).

[2771] 3594

[2772] 3595 Embodiment 419: The compound according to any one of embodiments 378 to 379 or 397 to

[2773] 3596 413, wherein in the second binding module said VHH antibody domain or fragment thereof

[2774] 3597 comprises a VHH antibody domain or fragment thereof according to (E) or (H).

[2775] 3598

[2776] 3599 Embodiment 420: The compound according to any one of embodiments 378 to 379 or 397 to

[2777] 3600 413, wherein in the second binding module said VHH antibody domain or fragment thereof

[2778] 3601 comprises a VHH antibody domain or fragment thereof according to (E).

[2779] 3602

[2780] 3603 Embodiment 421 : The compound according to any one of embodiments 378 to 379 or 397 to

[2781] 3604 413, wherein in the second binding module said VHH antibody domain or fragment thereof

[2782] 3605 comprises a VHH antibody domain or fragment thereof according to (F).

[2783] 3606

[2784] 3607 Embodiment 422: The compound according to any one of embodiments 375 to 421, wherein

[2785] 3608 said compound is a molecule.

[2786] 3609

[2787] 3610 Embodiment 423: The compound according to any one of embodiments 375 to 422, wherein

[2788] 3611 said compound comprises or is a protein.

[2789] 3612

[2790] 3613 By stating that the compound "comprises" a protein, the present disclosure designates that the

[2791] 3614 compound includes a part within its chemical structure that is a protein. A compound that

[2792] 3615 comprises a protein may or may not comprise a part that is not a protein.

[2793] 3616 3617 Embodiment 424: The compound according to any one of embodiments 375 to 422, wherein

[2794] 3618 said compound is a protein.

[2795] 3619

[2796] 3620 By stating that the compound "is" a protein, the present disclosure designates that the compound

[2797] 3621 consists only of protein and does not comprise a part that is not a protein.

[2798] 3622

[2799] 3623 Embodiment 425: The compound according to any one of embodiments 375 to 424, wherein

[2800] 3624 said compound further comprises a targeting moiety.

[2801] 3625

[2802] 3626 As used herein, the term "targeting moiety" refers to a moiety (i.e. a molecular group or

[2803] 3627 chemical structure) that is (typically covalently) associated with said compound and that binds

[2804] 3628 a target site, wherein said binding allows to recruit the compound to said target site (e.g. a

[2805] 3629 moiety that specifically binds to EGFR and thus targets the compound to cells expressing EGFR

[2806] 3630 at their cell surface). The target site will typically be a biological molecule or a certain part of

[2807] 3631 a biological molecule. An example of a targeting moiety is an antigen-binding antibody

[2808] 3632 fragment that is covalently linked to an IL-18Ra-binding and / or IL-18RP-binding VHH

[2809] 3633 antibody domain to form a compound according to the present disclosure, wherein the antigen¬

[2810] 3634 binding fragment binds to a certain receptor present at the surface of a certain cell type (its

[2811] 3635 antigen), and wherein binding of the antigen-binding fragment to this receptor results in

[2812] 3636 recruitment of the compound to this cell.

[2813] 3637

[2814] 3638 Non-targeted drugs typically reach their site of action by whole-body distribution and passive

[2815] 3639 diffusion. In contrast, targeted compounds are not distributed evenly across the whole body.

[2816] 3640 Due to the interaction of targeting moiety with its target molecule, a compound including a

[2817] 3641 targeting moiety is concentrated preferentially at its site target site. Therefore, e.g. therapeutic

[2818] 3642 compounds with a targeting moiety require lower dosages to be therapeutically effective, thus

[2819] 3643 improving the therapeutic window.

[2820] 3644

[2821] 3645 Embodiment 426: The compound according to embodiment 425, wherein said targeting moiety

[2822] 3646 is a protein, a peptide, a peptide mimetic, a nucleic acid, an oligonucleotide or a small molecule.

[2823] 3647

[2824] 3648 As used herein, the term "peptide mimetic" refers to a peptide-like chain which is designed to

[2825] 3649 mimic a peptide. An example of a peptide mimetic is a D-peptide mimetic containing a D- 3650 amino acid, but is not limited thereto. As used herein, a "small molecule", is a molecule with a

[2826] 3651 molecular weight < 1000 Da.

[2827] 3652

[2828] 3653 Embodiment 427: The compound according to any one of embodiments 375 to 426, wherein

[2829] 3654 said compound is a monospecific molecule.

[2830] 3655

[2831] 3656 Embodiment 428: The compound according to any one of embodiments 375 to 426, wherein

[2832] 3657 said compound is a bispecific molecule.

[2833] 3658

[2834] 3659 Embodiment 429: The compound according to any one of embodiments 375 to 426 or 428,

[2835] 3660 wherein said compound is a bispecific antibody molecule.

[2836] 3661

[2837] 3662 The term "bispecific antibody molecule", as used in the present disclosure, refers to an antibody

[2838] 3663 molecule that is capable of specifically binding to two different epitopes at the same time. This

[2839] 3664 does not exclude the possibility that said bispecific antibody molecule is linked to further

[2840] 3665 domains or moieties.

[2841] 3666

[2842] 3667 The terms "epitope" or "antigenic determinant" are used interchangeably herein and refer to the

[2843] 3668 portion of an antigen that is recognized and specifically bound by a particular antibody. When

[2844] 3669 the antigen is a polypeptide, epitopes can be formed both from contiguous amino acids and

[2845] 3670 noncontiguous amino acids juxtaposed by tertiary folding of a protein. Epitopes formed from

[2846] 3671 contiguous amino acids are typically retained upon protein denaturing, whereas epitopes

[2847] 3672 formed by tertiary folding are typically lost upon protein denaturing. An epitope typically

[2848] 3673 includes at least 3, and more usually, at least 5 or 8-10 amino acids in a unique spatial

[2849] 3674 conformation.

[2850] 3675

[2851] 3676 Typically, in the compounds of the present disclosure, one of the two epitopes to which the

[2852] 3677 compound of the present disclosure binds is an epitope on IL-18Ra whereas the other of the

[2853] 3678 two epitopes to which the compound of the present disclosure binds is an epitope on IL-18Rp.

[2854] 3679

[2855] 3680 Methods for making bispecific antibodies are known in the art. For example, bispecific

[2856] 3681 antibodies can be produced recombinantly using the co-expression of two immunoglobulin

[2857] 3682 heavy chain / light chain pairs (see e.g. Milstein et al., Nature (1983), vol. 305, p. 537-539).

[2858] 3683 Alternatively, bispecific antibodies can be prepared using chemical linkage (see e.g. Brennan 3684 et al., Science (1985), vol. 229, p. 81). A bispecific antibody can also for example be prepared

[2859] 3685 by the SEED technology (see below).

[2860] 3686

[2861] 3687 Embodiment 430: The compound according to any one of embodiments 375 to 426 or 428 to

[2862] 3688 429, wherein one binding site of said bispecific antibody molecule is formed by a VHH

[2863] 3689 antibody domain or fragment thereof according to the present disclosure that is specific for IL-

[2864] 3690 18Ra, and one binding site of said bispecific antibody is formed by a VHH antibody domain or

[2865] 3691 fragment thereof according to the present disclosure that is specific for IL-18Rp.

[2866] 3692

[2867] 3693 Embodiment 431: The compound according to any one of embodiments 375 to 426 or 428 to

[2868] 3694 429, wherein one binding site of said bispecific antibody molecule is a VHH antibody domain

[2869] 3695 or fragment thereof according to the present disclosure and the other binding site of said

[2870] 3696 bispecific antibody molecule is selected from the group consisting of a Fab, a Fab', a (Fab')2, a

[2871] 3697 Fv, a scFv, a diabody and a VHH.

[2872] 3698

[2873] 3699 "Fab" fragments are obtained by papain digestion of an antibody, which produces two identical

[2874] 3700 antigen-binding fragments, called "Fab" fragments, and a residual "Fc" fragment, a designation

[2875] 3701 reflecting the ability to crystallize readily. The Fab fragment consists of an entire L chain along

[2876] 3702 with the variable region domain of the H chain (VH), and the first constant domain of one heavy

[2877] 3703 chain (CHI). Each Fab fragment is monovalent with respect to antigen binding, i.e., it has a

[2878] 3704 single antigen-binding site.

[2879] 3705

[2880] 3706 "F(ab')2" fragments are obtained by pepsin treatment of an antibody, which yields a single large

[2881] 3707 F(ab')2 fragment which roughly corresponds to two disulfide linked Fab fragments having

[2882] 3708 different antigen-binding activity and is still capable of cross-linking antigen.

[2883] 3709

[2884] 3710 "Fab1" fragments differ from Fab fragments by having a few additional residues at the carboxy

[2885] 3711 terminus of the CHI domain including one or more cysteines from the antibody hinge region.

[2886] 3712 Fab'-SH is the designation for Fab' in which the cysteine residue(s) of the constant domains

[2887] 3713 bear a free thiol group. F(ab')2 antibody fragments originally were produced as pairs of Fab'

[2888] 3714 fragments which have hinge cysteines between them. Other chemical couplings of antibody

[2889] 3715 fragments are also known.

[2890] 3716 3717 The Fc fragment comprises the carboxy-terminal portions of both H chains held together by

[2891] 3718 disulfides. The effector functions of antibodies are determined by sequences in the Fc region,

[2892] 3719 the region which is also recognized by Fc receptors (FcR) found on certain types of cells.

[2893] 3720

[2894] 3721 "Fv" is the minimum antibody fragment which contains a complete antigen-recognition and -

[2895] 3722 binding site. This fragment consists of a dimer of one heavy- and one light-chain variable region

[2896] 3723 domain in tight, non-covalent association. From the folding of these two domains emanate six

[2897] 3724 hypervariable loops (3 loops each from the H and L chain) that contribute the amino acid

[2898] 3725 residues for antigen binding and confer antigen binding specificity to the antibody. However,

[2899] 3726 even a single variable domain (or half of an Fv comprising only three HVRs specific for an

[2900] 3727 antigen) has the ability to recognize and bind antigen, although at a lower affinity than the entire

[2901] 3728 binding site.

[2902] 3729

[2903] 3730 "Single-chain Fv", also abbreviated as "scFv", are antibody fragments that comprise the VH

[2904] 3731 and VL antibody domains connected into a single polypeptide chain. Preferably, the scFv

[2905] 3732 polypeptide further comprises a polypeptide linker between the VH and VL domains which

[2906] 3733 enables the scFv to form the desired structure for antigen binding. For a review of the scFv, see

[2907] 3734 Pluckthun, in: The Pharmacology of Monoclonal Antibodies, vol. 113 (1994), editors

[2908] 3735 Rosenburg and Moore, Springer-Verlag (New York), p. 269-315.

[2909] 3736

[2910] 3737 The term "diabody" refers to a small antibody fragment prepared by constructing scFv

[2911] 3738 fragments (see preceding paragraph) with short linkers (about 5-25 residues) between the VH

[2912] 3739 and VL domains such that inter-chain but not intra-chain pairing of the V domains is achieved,

[2913] 3740 thereby resulting in a bivalent fragment, i.e., a fragment having two antigen-binding sites.

[2914] 3741 Bispecific diabodies are heterodimers of two "crossover" scFv fragments in which the VH and

[2915] 3742 VL domains of the two antibodies are present on different polypeptide chains. Diabodies are

[2916] 3743 described in greater detail in, for example, EP 0404097; WO 93 / 11161; Hollinger et al., Proc.

[2917] 3744 Natl. Acad. Sci. USA (1993), vol. 90, p. 6444-6448.

[2918] 3745

[2919] 3746 Embodiment 432: The compound according to any one of embodiments 375 to 426 or 430 to

[2920] 3747 431, wherein said compound is a multispecific molecule.

[2921] 3748

[2922] 3749 Embodiment 433: The compound according to any one of embodiments 375 to 426 or 428 to

[2923] 3750 430, wherein said compound is a bispecific molecule that is specific for IL-18Ra and IL-18Rp. 3751

[2924] 3752 Embodiment 434: The compound according to any one of embodiments 375 to 426 or 428 to

[2925] 3753 430 or 433, wherein said compound is a bispecific molecule that binds to IL-18Ra and IL-

[2926] 3754 18R .

[2927] 3755

[2928] 3756 Embodiment 435: The compound according to any one of embodiments 375 to 426 or 428 to

[2929] 3757 430 or 433 to 434, wherein said compound is a bispecific molecule directed to IL-18Ra and IL-

[2930] 3758 18R .

[2931] 3759

[2932] 3760 Embodiment 436: The compound according to any one of embodiments 375 to 426 or 428 to

[2933] 3761 430 or 433 to 435, wherein said compound is a bispecific antibody molecule that is specific for

[2934] 3762 IL-18Ra and IL-18Rp.

[2935] 3763

[2936] 3764 If the present disclosure refers to "IL-18Ra" without indicating a specific organism from which

[2937] 3765 said IL-18Ra originates, then this refers to human IL-18Ra. If the present disclosure refers to

[2938] 3766 "IL-18RP" without indicating a specific organism from which said IL-18RP originates, then

[2939] 3767 this refers to human IL-18RP.

[2940] 3768

[2941] 3769 According to a twelfth aspect, the present disclosure relates to any of the following (To the

[2942] 3770 twelfth aspect of the present disclosure and the embodiments referring thereto, the same

[2943] 3771 explanations and definitions apply accordingly as for the first to eleventh aspects of the present

[2944] 3772 disclosure and the embodiments referring thereto.):

[2945] 3773

[2946] 3774 Embodiment 437: A compound which is a bispecific antibody molecule comprising an IL-

[2947] 3775 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 1 (First binding module) and

[2948] 3776 an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 12 (Second binding

[2949] 3777 module).

[2950] 3778

[2951] 3779 Embodiment 438: A compound which is a bispecific antibody molecule comprising an IL-

[2952] 3780 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 1 (First binding module) and

[2953] 3781 an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 13 (Second binding

[2954] 3782 module).

[2955] 3783 3784 Embodiment 439: A compound which is a bispecific antibody molecule comprising an IL-

[2956] 3785 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 1 (First binding module) and

[2957] 3786 an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 15 (Second binding

[2958] 3787 module).

[2959] 3788

[2960] 3789 Embodiment 440: A compound which is a bispecific antibody molecule comprising an IL-

[2961] 3790 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 1 (First binding module) and

[2962] 3791 an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 17 (Second binding

[2963] 3792 module).

[2964] 3793

[2965] 3794 Embodiment 441: A compound which is a bispecific antibody molecule comprising an IL-

[2966] 3795 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 1 (First binding module) and

[2967] 3796 an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 20 (Second binding

[2968] 3797 module).

[2969] 3798

[2970] 3799 Embodiment 442: A compound which is a bispecific antibody molecule comprising an IL-

[2971] 3800 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 1 (First binding module) and

[2972] 3801 an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 21 (Second binding

[2973] 3802 module).

[2974] 3803

[2975] 3804 Embodiment 443: A compound which is a bispecific antibody molecule comprising an IL-

[2976] 3805 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 1 (First binding module) and

[2977] 3806 an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 22 (Second binding

[2978] 3807 module).

[2979] 3808

[2980] 3809 Embodiment 444: A compound which is a bispecific antibody molecule comprising an IL-

[2981] 3810 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 2 (First binding module) and

[2982] 3811 an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 12 (Second binding

[2983] 3812 module).

[2984] 3813

[2985] 3814 Embodiment 445: A compound which is a bispecific antibody molecule comprising an IL-

[2986] 3815 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 2 (First binding module) and

[2987] 3816 an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 13 (Second binding

[2988] 3817 module). 3818

[2989] 3819 Embodiment 446: A compound which is a bispecific antibody molecule comprising an IL-

[2990] 3820 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 2 (First binding module) and

[2991] 3821 an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 15 (Second binding

[2992] 3822 module).

[2993] 3823

[2994] 3824 Embodiment 447: A compound which is a bispecific antibody molecule comprising an IL-

[2995] 3825 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 2 (First binding module) and

[2996] 3826 an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 17 (Second binding

[2997] 3827 module).

[2998] 3828

[2999] 3829 Embodiment 448: A compound which is a bispecific antibody molecule comprising an IL-

[3000] 3830 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 2 (First binding module) and

[3001] 3831 an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 18 (Second binding

[3002] 3832 module).

[3003] 3833

[3004] 3834 Embodiment 449: A compound which is a bispecific antibody molecule comprising an IL-

[3005] 3835 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 2 (First binding module) and

[3006] 3836 an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 20 (Second binding

[3007] 3837 module).

[3008] 3838

[3009] 3839 Embodiment 450: A compound which is a bispecific antibody molecule comprising an IL-

[3010] 3840 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 2 (First binding module) and

[3011] 3841 an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 21 (Second binding

[3012] 3842 module).

[3013] 3843

[3014] 3844 Embodiment 451: A compound which is a bispecific antibody molecule comprising an IL-

[3015] 3845 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 5 (First binding module) and

[3016] 3846 an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 13 (Second binding

[3017] 3847 module).

[3018] 3848

[3019] 3849 Embodiment 452: A compound which is a bispecific antibody molecule comprising an IL-

[3020] 3850 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 5 (First binding module) and 3851 an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 14 (Second binding

[3021] 3852 module).

[3022] 3853

[3023] 3854 Embodiment 453: A compound which is a bispecific antibody molecule comprising an IL-

[3024] 3855 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 6 (First binding module) and

[3025] 3856 an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 17 (Second binding

[3026] 3857 module).

[3027] 3858

[3028] 3859 Embodiment 454: A compound which is a bispecific antibody molecule comprising an IL-

[3029] 3860 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 6 (First binding module) and

[3030] 3861 an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 20 (Second binding

[3031] 3862 module).

[3032] 3863

[3033] 3864 Embodiment 455: A compound which is a bispecific antibody molecule comprising an IL-

[3034] 3865 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 6 (First binding module) and

[3035] 3866 an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 21 (Second binding

[3036] 3867 module).

[3037] 3868

[3038] 3869 Embodiment 456: A compound which is a bispecific antibody molecule comprising an IL-

[3039] 3870 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 6 (First binding module) and

[3040] 3871 an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 22 (Second binding

[3041] 3872 module).

[3042] 3873

[3043] 3874 Embodiment 457: A compound which is a bispecific antibody molecule comprising an IL-

[3044] 3875 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 8 (First binding module) and

[3045] 3876 an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 12 (Second binding

[3046] 3877 module).

[3047] 3878

[3048] 3879 Embodiment 458: A compound which is a bispecific antibody molecule comprising an IL-

[3049] 3880 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 8 (First binding module) and

[3050] 3881 an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 13 (Second binding

[3051] 3882 module).

[3052] 3883 3884 Embodiment 459: A compound which is a bispecific antibody molecule comprising an IL-

[3053] 3885 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 8 (First binding module) and

[3054] 3886 an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 17 (Second binding

[3055] 3887 module).

[3056] 3888

[3057] 3889 Embodiment 460: A compound which is a bispecific antibody molecule comprising an IL-

[3058] 3890 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 8 (First binding module) and

[3059] 3891 an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 18 (Second binding

[3060] 3892 module).

[3061] 3893

[3062] 3894 Embodiment 461: A compound which is a bispecific antibody molecule comprising an IL-

[3063] 3895 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 8 (First binding module) and

[3064] 3896 an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 20 (Second binding

[3065] 3897 module).

[3066] 3898

[3067] 3899 Embodiment 462: A compound which is a bispecific antibody molecule comprising an IL-

[3068] 3900 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 8 (First binding module) and

[3069] 3901 an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 21 (Second binding

[3070] 3902 module).

[3071] 3903

[3072] 3904 Embodiment 463: A compound which is a bispecific antibody molecule comprising an IL-

[3073] 3905 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 9 (First binding module) and

[3074] 3906 an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 12 (Second binding

[3075] 3907 module).

[3076] 3908

[3077] 3909 Embodiment 464: A compound which is a bispecific antibody molecule comprising an IL-

[3078] 3910 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 9 (First binding module) and

[3079] 3911 an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 13 (Second binding

[3080] 3912 module).

[3081] 3913

[3082] 3914 Embodiment 465: A compound which is a bispecific antibody molecule comprising an IL-

[3083] 3915 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 9 (First binding module) and

[3084] 3916 an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 15 (Second binding

[3085] 3917 module). 3918

[3086] 3919 Embodiment 466: A compound which is a bispecific antibody molecule comprising an IL-

[3087] 3920 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 9 (First binding module) and

[3088] 3921 an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 17 (Second binding

[3089] 3922 module).

[3090] 3923

[3091] 3924 Embodiment 467: A compound which is a bispecific antibody molecule comprising an IL-

[3092] 3925 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 9 (First binding module) and

[3093] 3926 an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 20 (Second binding

[3094] 3927 module).

[3095] 3928

[3096] 3929 Embodiment 468: A compound which is a bispecific antibody molecule comprising an IL-

[3097] 3930 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 9 (First binding module) and

[3098] 3931 an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 21 (Second binding

[3099] 3932 module).

[3100] 3933

[3101] 3934 Embodiment 469: A compound which is a bispecific antibody molecule comprising an IL-

[3102] 3935 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 9 (First binding module) and

[3103] 3936 an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 22 (Second binding

[3104] 3937 module).

[3105] 3938

[3106] 3939 Embodiment 470: A compound which is a bispecific antibody molecule comprising an IL-

[3107] 3940 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 10 (First binding module)

[3108] 3941 and an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 13 (Second binding

[3109] 3942 module).

[3110] 3943

[3111] 3944 Embodiment 471 : A compound which is a bispecific antibody molecule comprising an IL-

[3112] 3945 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 10 (First binding module)

[3113] 3946 and an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 20 (Second binding

[3114] 3947 module).

[3115] 3948

[3116] 3949 Embodiment 472: A compound which is a bispecific antibody molecule comprising an IL-

[3117] 3950 18Ra-binding VHH with the amino acid sequence of SEQ ID NO: 10 (First binding module) 3951 and an IL18RP-binding VHH with the amino acid sequence of SEQ ID NO: 21 (Second binding

[3118] 3952 module).

[3119] 3953

[3120] 3954 Embodiment 473: The compound according to any one of embodiments 375 or 377 to 472,

[3121] 3955 wherein said compound is monovalent for binding to IL-18Ra.

[3122] 3956

[3123] 3957 Embodiment 474: The compound according to any one of embodiments 375 or 377 to 472,

[3124] 3958 wherein said compound is bivalent for binding to IL-18Ra.

[3125] 3959

[3126] 3960 Embodiment 475: The compound according to any one of embodiments 375 or 377 to 472,

[3127] 3961 wherein said compound is multivalent for binding to IL-18Ra.

[3128] 3962

[3129] 3963 Embodiment 476: The compound according to any one of embodiments 376 to 475, wherein

[3130] 3964 said compound is monovalent for binding to IL-18RP.

[3131] 3965

[3132] 3966 Embodiment 477: The compound according to any one of embodiments 376 to 475, wherein

[3133] 3967 said compound is bivalent for binding to IL-18RP.

[3134] 3968

[3135] 3969 Embodiment 478: The compound according to any one of embodiments 376 to 475, wherein

[3136] 3970 said compound is multivalent for binding to IL-18RP.

[3137] 3971

[3138] 3972 Embodiment 479: The compound according to any one of embodiments 377 to 478, wherein

[3139] 3973 said compound comprises one copy of said first binding module.

[3140] 3974

[3141] 3975 Embodiment 480: The compound according to any one of embodiments 377 to 478, wherein

[3142] 3976 said compound comprises two copies of said first binding module.

[3143] 3977

[3144] 3978 Embodiment 481: The compound according to any one of embodiments 377 to 478, wherein

[3145] 3979 said compound comprises two or more copies of said first binding module.

[3146] 3980

[3147] 3981 Embodiment 482: The compound according to any one of embodiments 377 to 479, wherein

[3148] 3982 said compound comprises not more than one copy of said first binding module.

[3149] 3983 3984 Embodiment 483: The compound according to any one of embodiments 377 to 478 or 480,

[3150] 3985 wherein said compound comprises not more than two copies of said first binding module.

[3151] 3986

[3152] 3987 Embodiment 484: The compound according to any one of embodiments 377 to 483, wherein

[3153] 3988 said compound comprises one copy of said second binding module.

[3154] 3989

[3155] 3990 Embodiment 485: The compound according to any one of embodiments 377 to 483, wherein

[3156] 3991 said compound comprises two copies of said second binding module.

[3157] 3992

[3158] 3993 Embodiment 486: The compound according to any one of embodiments 377 to 483, wherein

[3159] 3994 said compound comprises two or more copies of said second binding module.

[3160] 3995

[3161] 3996 Embodiment 487: The compound according to any one of embodiments 377 to 484, wherein

[3162] 3997 said compound comprises not more than one copy said second binding module.

[3163] 3998

[3164] 3999 Embodiment 488: The compound according to any one of embodiments 377 to 483 or 485,

[3165] 4000 wherein said compound comprises not more than two copies of said second binding module.

[3166] 4001

[3167] 4002 Embodiment 489: The compound according to any one of embodiments 375 to 488, wherein

[3168] 4003 said compound comprises only one molecular structure that binds to IL-18Ra.

[3169] 4004

[3170] 4005 Embodiment 490: The compound according to any one of embodiments 375 to 489, wherein

[3171] 4006 said compound comprises only one molecular structure that binds to IL-18RP.

[3172] 4007

[3173] 4008 Embodiment 491: The compound according to any one of embodiments 375 to 490, wherein

[3174] 4009 all components of said compound are covalently linked.

[3175] 4010

[3176] 4011 Embodiment 492: The compound according to any one of embodiments 377 to 491, wherein

[3177] 4012 said first binding module is a VHH antibody domain. As the skilled person understands, by

[3178] 4013 stating that the first binding module "is a VHH antibody domain", the present disclosure

[3179] 4014 indicates that the first binding module is a full VHH antibody domain, not just a fragment

[3180] 4015 thereof.

[3181] 4016 4017 Embodiment 493: The compound according to any one of embodiments 377 to 492, wherein

[3182] 4018 said second binding module is a VHH antibody domain.

[3183] 4019

[3184] 4020 Embodiment 494: The compound according to any one of embodiments 377 to 491, wherein

[3185] 4021 said first binding module consists of a fragment of a VHH antibody domain. As the skilled

[3186] 4022 person understands, by stating that the first binding module "consists of a fragment of a VHH

[3187] 4023 antibody domain", the present disclosure indicates that the first binding module is a fragment

[3188] 4024 of a VHH antibody domain, not a full-length VHH antibody domain.

[3189] 4025

[3190] 4026 Embodiment 495: The compound according to any one of embodiments 377 to 491 or 494,

[3191] 4027 wherein said second binding module consists of a fragment of a VHH antibody domain.

[3192] 4028

[3193] 4029 Embodiment 496: The compound according to any one of embodiments 375 to 495, wherein

[3194] 4030 said compound comprises an antibody Fc region.

[3195] 4031

[3196] 4032 As used herein, the term "antibody Fc region" refers to the portion of a native immunoglobulin

[3197] 4033 formed by the Fc domains of its two heavy chains (which includes a heavy chain constant region

[3198] 4034 1 (CHI), a heavy chain constant region 2 (CH2) and a heavy chain constant region 3 (CH3) of

[3199] 4035 an immunoglobulin, but does not include variable regions of the heavy and light chains and a

[3200] 4036 light chain constant region 1 (CL1) of an immunoglobulin). A native Fc region is homodimeric.

[3201] 4037 In some embodiments, the term includes variant Fc regions with one or more alterations relative

[3202] 4038 to a native Fc region. An Fc region may be altered by amino acid substitutions, additions and / or

[3203] 4039 deletions, linkage of additional moieties, and / or alteration of the native glycans. The term

[3204] 4040 encompasses Fc regions wherein each of the constituent Fc domains is different. Examples of

[3205] 4041 heterodimeric Fc regions include, without limitation, Fc regions made using the "knobs into

[3206] 4042 holes" technology as described in, for example US Patent No. 8,216,805 or by the SEED

[3207] 4043 technology as described in WO 2016 / 087650.

[3208] 4044

[3209] 4045 Embodiment 497: The compound according to any one of embodiments 375 to 495, wherein

[3210] 4046 said compound comprises an antibody Fc region that is competent in Fc receptor binding.

[3211] 4047

[3212] 4048 An antibody Fc region is "competent in FC receptor binding" if said antibody Fc region is

[3213] 4049 capable of binding to at least one of the Fc receptors (FcyRI, FcyRII, and FcyRIII subclasses,

[3214] 4050 including allelic variants and alternatively spliced forms of these receptors). 4051

[3215] 4052 Embodiment 498: The compound according to any one of embodiments 375 to 495, wherein

[3216] 4053 said compound comprises an antibody Fc region that is not competent in Fc receptor binding.

[3217] 4054

[3218] 4055 Embodiment 499: The compound according to any one of embodiments 375 to 495, wherein

[3219] 4056 said compound does not comprise an effector-competent antibody Fc region.

[3220] 4057

[3221] 4058 An "effector-competent" Fc region is an Fc region having the functional ability to bind proteins

[3222] 4059 and / or cells of the immune system and mediate biological effects normally induced following

[3223] 4060 the binding of an antibody to a corresponding antigen. Such biological effects include e.g. the

[3224] 4061 ability to bind a complement protein (e.g. Clq), resulting in activation of the classical

[3225] 4062 complement system leading to the opsonisation and lysis of cell pathogens (complement¬

[3226] 4063 dependent cytotoxicity, CDCC). Other biological effects are endocytosis of immune

[3227] 4064 complexes, engulfment and destruction of antibody-coated particles or microorganisms (also

[3228] 4065 called antibody-dependent phagocytosis, or ADCP), clearance of immune complexes, lysis of

[3229] 4066 antibody-coated target cells by killer cells (called antibody-dependent cell-mediated

[3230] 4067 cytotoxicity, or ADCC), release of inflammatory mediators, regulation of immune system cell

[3231] 4068 activation or control of immunoglobulin production.

[3232] 4069

[3233] 4070 Embodiment 500: The compound according to any one of embodiments 375 to 497, wherein

[3234] 4071 said compound comprises an effector-competent antibody Fc region.

[3235] 4072

[3236] 4073 Embodiment 501 : The compound according to any one of embodiments 375 to 495 or 498 to

[3237] 4074 499, wherein said compound does not comprise an antibody Fc region capable of inducing

[3238] 4075 ADCC (antibody-dependent cellular cytotoxicity).

[3239] 4076

[3240] 4077 Embodiment 502: The compound according to any one of embodiments 375 to 497 or 500,

[3241] 4078 wherein said compound comprises an antibody Fc region capable of inducing ADCC.

[3242] 4079

[3243] 4080 Embodiment 503: The compound according to any one of embodiments 375 to 499, wherein

[3244] 4081 said compound comprises an antibody Fc region that is not capable of inducing ADCC.

[3245] 4082

[3246] 4083 Embodiment 504: The compound according to any one of embodiments 375 to 503, wherein

[3247] 4084 that antibody Fc region carries a mutation E430G. 4085

[3248] 4086 This mutation is described in de Jong et al., 2016 and has been reported to drive antibody

[3249] 4087 hexamerization upon target cell binding.

[3250] 4088

[3251] 4089 Embodiment 505: The compound according to any one of embodiments 377 to 504, wherein

[3252] 4090 said compound is a bispecific antibody molecule prepared by the SEED technology (i.e. a

[3253] 4091 SEEDbody).

[3254] 4092

[3255] 4093 The SEED (strand-exchange engineered domain) technology is an approach for generation of

[3256] 4094 bispecific antibodies in which structurally related sequences within the conserved CH3 domains

[3257] 4095 of human IgA and IgG are exchanged to form two asymmetric but complementary domains

[3258] 4096 (see the Examples section). For details about the SEED technology are described in WO

[3259] 4097 2016 / 087650 and Davis et al., 2010. A bispecific antibody prepared by the SEED technology

[3260] 4098 is herein also referred to as "SEEDbody".

[3261] 4099

[3262] 4100 Embodiment 506: The compound according to any one of embodiments 377 to 505, wherein

[3263] 4101 the VHH antibody domain or fragment thereof of the first binding module is formed by a

[3264] 4102 sequence selected from the group consisting of SEQ ID NO: 89, SEQ ID NO: 90, SEQ ID NO:

[3265] 4103 91, SEQ ID NO: 92, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 96, SEQ ID NO: 97, SEQ

[3266] 4104 ID NO: 98 and SEQ ID NO: 99,

[3267] 4105 and wherein the VHH antibody domain or fragment thereof of the second binding module is

[3268] 4106 formed by a sequence selected from the group consisting of SEQ ID NO: 100, SEQ ID NO:

[3269] 4107 101, SEQ ID NO: 102, SEQ ID NO: 103, SEQ ID NO: 104, SEQ ID NO: 105, SEQ ID NO:

[3270] 4108 106, SEQ ID NO: 107, SEQ ID NO: 108, SEQ ID NO: 109 and SEQ ID NO: 110.

[3271] 4109

[3272] 4110 SEQ ID NO: 89 to 99 define the amino acid sequences of SEED AG chains based on VHH1 to

[3273] 4111 VHH11 (al to pi l). SEQ ID NO: 100 to 110 define the amino acid sequences of SEED GA

[3274] 4112 chains based on VHH12 to VHH22 (pi2 to P22).

[3275] 4113

[3276] 4114 Embodiment 507: The compound according to any one of embodiments 377 to 506, wherein

[3277] 4115 said compound is a bispecific antibody with a strictly monovalent (1+1) bispecific sdAb (single¬

[3278] 4116 domain antibody) architecture.

[3279] 4117 4118 Embodiment 508: The compound according to any one of embodiments 377 to 507, wherein

[3280] 4119 said compound comprises two VHHs of different binding specificity, and each of these VHHs

[3281] 4120 is present in said compound in only one copy. As the skilled person understands, if the

[3282] 4121 compound is a bispecific antibody comprising a first and a second binding module, then

[3283] 4122 typically one VHH forms the first binding module of said compound whereas the other VHH

[3284] 4123 forms the second binding module of said compound.

[3285] 4124

[3286] 4125 Embodiment 509: The compound according to any one of embodiments 377 to 504 or 506 to

[3287] 4126 508, wherein said compound is a single domain-based IgG (sdlgG).

[3288] 4127

[3289] 4128 Embodiment 510: The compound according to embodiment 509, wherein a sdlgG is a

[3290] 4129 conventional IgG antibody in which the two VH regions are replaced by VHH domains of one

[3291] 4130 binding specificity, and the two VL regions are replaced by VHH domains of a different binding

[3292] 4131 specificity, resulting in a bispecific, tetravalent antibody.

[3293] 4132

[3294] 4133 The single domain-based IgG (sdlgG) format is described in detail in Pekar et al., 2020.

[3295] 4134

[3296] 4135 Embodiment 511: The compound according to any one of embodiments 509 to 510, wherein

[3297] 4136 said sdlgG is a conventional IgG antibody in which each of the two VH regions is replaced by

[3298] 4137 a copy of said first binding module, and each of the two VL regions is replaced by a copy of

[3299] 4138 said second binding module, resulting in a bispecific, tetravalent antibody.

[3300] 4139

[3301] 4140 Embodiment 512: The compound according to embodiment 511, wherein the first binding

[3302] 4141 module is a VHH that has been grafted onto the CHI domain of the heavy chain of an IgGl and

[3303] 4142 the second binding module is a VHH that has been grafted onto the constant region of the

[3304] 4143 lambda light chain of an IgGl .

[3305] 4144

[3306] 4145 Embodiment 513: The compound according to any one of embodiments 510 to 512, wherein

[3307] 4146 said IgG / IgGl is a human IgGl.

[3308] 4147

[3309] 4148 Embodiment 514: The compound according to any one of embodiments 377 to 504 or 507 to

[3310] 4149 to 513, wherein the IL-18Ra-specific VHH2 (SEQ ID NO: 2) has been grafted onto the CHI

[3311] 4150 domain of the heavy chain of an IgGl and the IL-18RP-specific VHH15 (SEQ ID NO: 15) has

[3312] 4151 been grafted onto the constant region of the lambda light chain of an IgGl. 4152

[3313] 4153 Embodiment 515: The compound according to any one of embodiments 377 to 514, wherein

[3314] 4154 said heavy chain of said IgG is effector-silenced.

[3315] 4155

[3316] 4156 Embodiment 516: The compound according to any one of embodiments 377 to 504 or 515,

[3317] 4157 wherein said compound consists of a IgG Fc region to which two copies of a tandem-VHH

[3318] 4158 arrangement are covalently linked at the hinge-region, wherein said tandem-VHH arrangement

[3319] 4159 consists of a first VHH antibody domain that is covalently linked to a second VHH antibody

[3320] 4160 domain, wherein said second VHH antibody domain is covalently linked to the hinge region of

[3321] 4161 said Fc region, resulting in a bispecific, tetravalent compound.

[3322] 4162

[3323] 4163 Embodiment 517: The compound according to embodiment 516, wherein said IgG Fc region is

[3324] 4164 an effector-silenced IgGl Fc region.

[3325] 4165

[3326] 4166 Embodiment 518: The compound according to any one of embodiments 516 to 517, wherein

[3327] 4167 said first binding module forms said first VHH antibody domain and said second binding

[3328] 4168 module forms said second VHH antibody domain.

[3329] 4169

[3330] 4170 Embodiment 519: The compound according to any one of embodiments 516 to 518, wherein

[3331] 4171 said first VHH antibody domain and said second VHH antibody domain are separated by a

[3332] 4172 linker.

[3333] 4173

[3334] 4174 Embodiment 520: The compound according to any one of embodiments 516 to 519, wherein

[3335] 4175 said first VHH antibody domain and said second VHH antibody domain are separated by a five

[3336] 4176 amino acid Gly4Ser-linker.

[3337] 4177

[3338] 4178 Embodiment 521: The compound according to any one of embodiments 516 to 520, wherein

[3339] 4179 said first VHH antibody domain is specific for IL-18Ra and said second VHH antibody domain

[3340] 4180 is specific for IL-18RP.

[3341] 4181

[3342] 4182 Embodiment 522: The compound according to any one of embodiments 516 to 521, wherein

[3343] 4183 said first VHH antibody domain has the amino acid sequence of VHH2 (SEQ ID NO: 2) and

[3344] 4184 said second VHH antibody domain has the amino acid sequence of VHH15 (SEQ ID NO: 15).

[3345] 4185 4186 Embodiment 523: The compound according to any one of embodiments 516 to 520, wherein

[3346] 4187 said first VHH antibody domain is specific for IL-18RP and said second VHH antibody domain

[3347] 4188 is specific for IL-18Ra.

[3348] 4189

[3349] 4190 Embodiment 524: The compound according to any one of embodiments 516 to 520 or 523,

[3350] 4191 wherein said first VHH antibody domain has the amino acid sequence of VHH15 (SEQ ID NO:

[3351] 4192 15) and said second VHH antibody domain has the amino acid sequence of VHH2 (SEQ ID

[3352] 4193 NO: 2).

[3353] 4194

[3354] 4195 Embodiment 525: The compound according to any one of embodiments 516 to 524, wherein

[3355] 4196 the orientation of the first and second VHH antibody domains is the first VHH antibody

[3356] 4197 domains followed by the second VHH antibody domain (from N-terminus to C-terminus).

[3357] 4198

[3358] 4199 Embodiment 526: The compound according to any one of embodiments 516 to 525, wherein

[3359] 4200 the orientation of the first and second VHH antibody domains is the second VHH antibody

[3360] 4201 domains followed by the first VHH antibody domain (from N-terminus to C-terminus).

[3361] 4202

[3362] 4203 Embodiment 527: The compound according to any one of embodiments 375 to 526, wherein

[3363] 4204 said compound is an IL-18 mimetic.

[3364] 4205

[3365] 4206 As used herein, the term "mimetic" refers to a compound that imitates certain aspects of the

[3366] 4207 function of another compound. Provided herein are, among other things, IL-18 mimetics. IL-

[3367] 4208 18 is a proinflammatory cytokine belonging to the IL-1 family of cytokines that mediates

[3368] 4209 signalling through heterodimerization of the receptor subunits IL-18Ra and IL-18RP (Yasuda

[3369] 4210 et al., 2019; Dinarello et al., 2013). IL-18 stimulates IFN-y production in innate lymphoid cells

[3370] 4211 as well as antigen-experienced T cells in synergy with IL-12. The aspects of the function of

[3371] 4212 IL 18 that such IL- 18 mimetics imitate is the induction of heterodimerization of IL-18R (from

[3372] 4213 IL-18Ra and IL-18RP) in the presence of IL-12 (or IL-15). Without being bound by theory, it

[3373] 4214 is speculated that by binding to both IL-18Ra and IL-18RP, the IL-18 mimetics of the present

[3374] 4215 disclosure crosslink IL-18Ra and IL-18RP, thus leading to dimerization and activation of IL-

[3375] 4216 18R in the presence of IL-12 (or IL-15). Thus, the IL-18 mimetics of the present disclosure

[3376] 4217 imitate the biological function of IL-18, for example the function of IL-18 to induce IFN-y

[3377] 4218 release (in the presence of IL-12 or IL-15).

[3378] 4219 4220 Embodiment 528: The compound according to embodiment 527, wherein said IL- 18 mimetic

[3379] 4221 is not affected from inhibition by IL-18BP (IL-18 binding protein).

[3380] 4222

[3381] 4223 Embodiment 529: The compound according to any one of embodiments 375 to 528, wherein

[3382] 4224 said compound is a IL-18 receptor (IL-18R) agonist.

[3383] 4225

[3384] 4226 Embodiment 530: The compound according to any one of embodiments 375 to 529, wherein

[3385] 4227 said compound is a cytokine mimetic targeting IL-18Ra and IL-18RP.

[3386] 4228

[3387] 4229 Embodiment 531: The compound according to any one of embodiments 375 to 530, wherein

[3388] 4230 said compound binds to IL-18Ra.

[3389] 4231

[3390] 4232 Embodiment 532: The compound according to any one of embodiments 375 to 531, wherein

[3391] 4233 said compound specifically binds to IL-18Ra.

[3392] 4234

[3393] 4235 Embodiment 533: The compound according to any one of embodiments 375 to 532, wherein

[3394] 4236 said compound is capable of binding to IL-18Ra with an affinity that is at least equal to the

[3395] 4237 affinity with which VHH1 (SEQ ID NO: 1), VHH2 (SEQ ID NO: 2), VHH3 (SEQ ID NO: 3),

[3396] 4238 VHH4 (SEQ ID NO: 4), VHH5 (SEQ ID NO: 5), VHH6 (SEQ ID NO: 6), VHH8 (SEQ ID NO:

[3397] 4239 8), VHH9 (SEQ ID NO: 9) or VHH10 (SEQ ID NO: 10) binds to IL-18Ra (or stronger).

[3398] 4240

[3399] 4241 Embodiment 534: The compound according to any one of embodiments 375 to 533, wherein

[3400] 4242 said compound binds to IL-18RP.

[3401] 4243

[3402] 4244 Embodiment 535: The compound according to any one of embodiments 375 to 534, wherein

[3403] 4245 said compound specifically binds to IL-18RP.

[3404] 4246

[3405] 4247 Embodiment 536: The compound according to any one of embodiments 375 to 535, wherein

[3406] 4248 said compound is capable of binding to IL-18RP with an affinity that is at least equal to the

[3407] 4249 affinity with which VHH12 (SEQ ID NO: 12), VHH13 (SEQ ID NO: 13), VHH14 (SEQ ID

[3408] 4250 NO: 14), VHH15 (SEQ ID NO: 15), VHH16 (SEQ ID NO: 16), VHH17 (SEQ ID NO: 17),

[3409] 4251 VHH18 (SEQ ID NO: 18), VHH20 (SEQ ID NO: 20), VHH21 (SEQ ID NO: 21) or VHH22

[3410] 4252 (SEQ ID NO: 22) binds to IL-18RP (or stronger).

[3411] 4253 4254 Embodiment 537: The compound according to any one of embodiments 375 to 536, wherein

[3412] 4255 said compound binds to IL-18Ra and IL-18RP.

[3413] 4256

[3414] 4257 Embodiment 538: The compound according to any one of embodiments 375 to 537, wherein

[3415] 4258 said compound specifically binds to IL-18Ra and IL-18Rp.

[3416] 4259

[3417] 4260 Embodiment 539: The compound according to any one of embodiments 375 to 538, wherein

[3418] 4261 said compound is capable of crosslinking IL-18Ra and IL-18RP.

[3419] 4262

[3420] 4263 Embodiment 540: The compound according to any one of embodiments 375 to 539, wherein

[3421] 4264 said compound is capable of inducing heterodimerization of IL-18Ra and IL-18RP.

[3422] 4265

[3423] 4266 Such binding / crosslinking / heterodimerization can be determined by in vitro binding

[3424] 4267 experiments as described in Example 1 below (by biolayer interferometry).

[3425] 4268

[3426] 4269 Embodiment 541: The compound according to any one of embodiments 375 to 540, wherein

[3427] 4270 said compound binds to IL-18Ra ECD with a KD of 100 nM or stronger.

[3428] 4271

[3429] 4272 Embodiment 542: The compound according to any one of embodiments 375 to 540, wherein

[3430] 4273 said compound binds to IL-18Ra ECD with a KD of 50 nM or stronger.

[3431] 4274

[3432] 4275 Embodiment 543: The compound according to any one of embodiments 375 to 540, wherein

[3433] 4276 said compound binds to IL-18Ra ECD with a KD of 25 nM or stronger.

[3434] 4277

[3435] 4278 Embodiment 544: The compound according to any one of embodiments 375 to 540, wherein

[3436] 4279 said compound binds to IL-18Ra ECD with a KD of 10 nM or stronger.

[3437] 4280

[3438] 4281 Embodiment 545: The compound according to any one of embodiments 375 to 540, wherein

[3439] 4282 said compound binds to IL-18Ra ECD with a KD of 1 nM or stronger.

[3440] 4283

[3441] 4284 Embodiment 546: The compound according to any one of embodiments 375 to 540, wherein

[3442] 4285 said c...

Claims

CLAIMS1. A compound comprising- a first binding module which is a VHH antibody domain or fragment thereof, wherein(a) said VHH antibody domain or fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of one VHH selected from the group consisting of VHH1, VHH2, VHH5, VHH6, VHH8, VHH9 and VHH10 as shown in the Table of CDRs;(b) said VHH antibody domain or fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 as defined in (a) with modification, wherein the modification is that the sequence of at least one of CDR1, CDR2 and CDR3 is humanized; or(c) said VHH antibody domain or fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 as defined in (a) with modification, wherein the modification is- the replacement, addition or deletion of up to three amino acids in CDR1,- the replacement, addition or deletion of up to three amino acids in CDR2 and / or- the replacement, addition or deletion of up to three amino acids in CDR3;- a second binding module which is a VHH antibody domain or a fragment thereof, wherein(d) said VHH antibody domain or fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of one VHH selected from the group consisting of VHH12, VHH13, VHH14, VHH15, VHH17, VHH18, VHH20, VHH21 and VHH22 as shown in the Table of CDRs, wherein said VHH antibody domain or fragment thereof defined in (d) is selected as follows: if the VHH antibody domain of (a) or the fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of VHH1, then the VHH antibody domain of (d) or the fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of one VHH selected from the group consisting of VHH12,VHH13, VHH15, VHH17, VHH20, VHH21 and VHH22 as shown in the Table of CDRs; if the VHH antibody domain of (a) or the fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of VHH2, then the VHH antibody domain of (d) or the fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of one VHH selected from the group consisting of VHH12, VHH13, VHH15, VHH17, VHH18, VHH20, VHH21 and VHH22 as shown in the Table of CDRs; if the VHH antibody domain of (a) or the fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of VHH5, then the VHH antibody domain of (d) or the fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of one VHH selected from the group consisting of VHH13 and VHH14 as shown in the Table of CDRs; if the VHH antibody domain of (a) or the fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of VHH6, then the VHH antibody domain of (d) or the fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of one VHH selected from the group consisting of VHH17, VHH20, VHH21 and VHH22 as shown in the Table of CDRs; if the VHH antibody domain of (a) or the fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of VHH8, then the VHH antibody domain of (d) or the fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of one VHH selected from the group consisting of VHH12, VHH13, VHH17, VHH18, VHH20 and VHH21 as shown in the Table of CDRs; if the VHH antibody domain of (a) or the fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of VHH9, then the VHH antibody domain of (d) or the fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of one VHH selected from the group consisting of VHH12,VHH13, VHH15, VHH17, VHH18, VHH20, VHH21 and VHH22 as shown in the Table of CDRs; if the VHH antibody domain of (a) or the fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of VHH10, then the VHH antibody domain of (d) or the fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 of one VHH selected from the group consisting of VHH13, VHH20 and VHH21 as shown in the Table of CDRs;(e) said VHH antibody domain or fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 as defined in (d) with modification, wherein the modification is that the sequence of at least one of CDR1, CDR2 and CDR3 is humanized; or(f) said VHH antibody domain or fragment thereof comprises the complementarity determining regions CDR1, CDR2 and CDR3 as defined in (d) with modification, wherein the modification is- the replacement, addition or deletion of up to one amino acid in CDR1 ,- the replacement, addition or deletion of up to one amino acid in CDR2 and / or- the replacement, addition or deletion of up to one amino acid in CDR3;Table of CDRs:

2. The compound according to claim 1, wherein said VHH antibody domain or fragment thereof defined in (a) comprises the complementarity determining regions CDR1, CDR2 and CDR3 of one VHH selected from the group consisting of VHH2 and VHH8 as shown in the Table of CDRs and wherein said VHH antibody domain or fragment thereof defined in (d) comprises the complementarity determining regions CDR1, CDR2 and CDR3 of one VHH selected from the group consisting of VHH15 and VHH17 as shown in the Table of CDRs.

3. The compound according to claim 1, wherein said VHH antibody domain or fragment thereof defined in (a) comprises the complementarity determining regions CDR1, CDR2 and CDR3 of VHH2 as shown in the Table of CDRs and wherein said VHH antibody domain or fragment thereof defined in (d) comprises the complementarity determining regions CDR1, CDR2 and CDR3 of VHH15 as shown in the Table of CDRs.

4. A compound comprising- a first binding module which is a VHH antibody domain or fragment thereof, wherein(A) said VHH antibody domain comprises the amino acid sequence of a VHH selected from the group consisting of VHH1, VHH2, VHH5, VHH6, VHH8, VHH9 and VHH10 as shown in the Table of VHH Sequences;(B) said VHH antibody domain comprises a VHH sequence as defined in (A) with modification, wherein the modification is that said sequence is humanized;(C) said VHH antibody domain comprises a VHH sequence as defined in (A) with modification, wherein the modification is the replacement, addition or deletion of up to 25 amino acids; or(D) said VHH antibody domain comprises a VHH sequence that is at least 75% identical to a VHH sequence referred to in (A);- a second binding module which is a VHH antibody domain or a fragment thereof, wherein(E) said VHH antibody domain comprises the amino acid sequence of a VHH selected from the group consisting of VHH12, VHH13, VHH14, VHH15, VHH17, VHH18, VHH20, VHH21 and VHH22 as shown in the Table of VHH Sequences, wherein said VHH antibody domain of (E) is selected as follows: if the VHH antibody domain of (A) comprises the amino acid sequence of VHH1, then the VHH antibody domain of (E) comprises the amino acid sequence of one VHH selected from the group consisting of VHH12, VHH13, VHH15, VHH17, VHH20, VHH21 and VHH22 shown in the Table of VHH Sequences; if the VHH antibody domain of (A) comprises the amino acid sequence of VHH2, then the VHH antibody domain of (E) comprises the amino acid sequence of one VHH selected from the group consisting of VHH12,VHH13, VHH15, VHH17, VHH18, VHH20, VHH21 and VHH22 shown in the Table of VHH Sequences; if the VHH antibody domain of (A) comprises the amino acid sequence of VHH5, then the VHH antibody domain of (E) comprises the amino acid sequence of one VHH selected from the group consisting of VHH13 and VHH14 shown in the Table of VHH Sequences; if the VHH antibody domain of (A) comprises the amino acid sequence of VHH6, then the VHH antibody domain of (E) comprises the amino acid sequence of one VHH selected from the group consisting of VHH17, VHH20, VHH21 and VHH22 shown in the Table of VHH Sequences; if the VHH antibody domain of (A) comprises the amino acid sequence of VHH8, then the VHH antibody domain of (E) comprises the amino acid sequence of one VHH selected from the group consisting of VHH12, VHH13, VHH17, VHH18, VHH20 and VHH21 shown in the Table of VHH Sequences; if the VHH antibody domain of (A) comprises the amino acid sequence of VHH9, then the VHH antibody domain of (E) comprises the amino acid sequence of one VHH selected from the group consisting of VHH12, VHH13, VHH15, VHH17, VHH18, VHH20, VHH21 and VHH22 shown in the Table of VHH Sequences; if the VHH antibody domain of (A) comprises the amino acid sequence of VHH10, then the VHH antibody domain of (E) comprises the amino acid sequence of one VHH selected from the group consisting of VHH13, VHH20 and VHH21 shown in the Table of VHH Sequences;(F) said VHH antibody domain comprises a VHH sequence as defined in (E) with modification, wherein the modification is that said sequence is humanized;(G) said VHH antibody domain comprises a VHH sequence as defined in (E) with modification, wherein the modification is the replacement, addition or deletion of up to 10 amino acids; or(H) said VHH antibody domain comprises a VHH sequence that is at least 90% identical to a VHH sequence referred to in (E);Table of VHH Sequences:

5. The compound according to claim 4, wherein said VHH antibody domain of (A) comprises the amino acid sequence of one VHH selected from the group consisting of VHH2 and VHH8 shown in the Table of VHH Sequences and wherein said VHH antibody domain of (E) comprises the amino acid sequence of one VHH selected from the group consisting of VHH15 and VHH17 shown in the Table of VHH Sequences.

6. The compound according to claim 4, wherein said VHH antibody domain of (A) comprises the amino acid sequence of VHH2 shown in the Table of VHH Sequences and wherein said VHH antibody domain of (E) comprises the amino acid sequence of VHH15 shown in the Table of VHH Sequences.

7. The compound according to any one of claims 1 to 6, wherein said compound is a bispecific antibody molecule prepared by the SEED technology.

8. The compound according to any one of claims 1 to 6, wherein said compound is a single domain-based IgG (sdlgG), wherein said sdlgG is a conventional IgG antibody in which each of the two VH regions is replaced by a copy of said first binding module, and each of the two VL regions is replaced by a copy of said second binding module, resulting in a bispecific, tetravalent antibody.

9. The compound according to any one of claims 1 to 6, wherein said compound consists of a IgG Fc region to which two copies of a tandem -VHH arrangement are covalently linked at the hinge-region, wherein said tandem-VHH arrangement consists of a first VHH antibody domain that is covalently linked to a second VHH antibody domain, wherein said second VHH antibody domain is covalently linked to the hinge region of said Fc region, resulting in a bispecific, tetravalent compound.

10. The compound according to any one of claims 1 to 9, wherein said compound binds to IL-18Ra ECD with a KD of 100 nM or stronger.

11. The compound according to any one of claims 1 to 10, wherein said compound binds to IL-18RP ECD with a KD of 100 nM or stronger.

12. The compound according to any one of claims 1 to 11, wherein said compound is capable of activating the IL-18 receptor IL-18R by binding to IL-18Ra and IL-18RP.

13. The compound according to any one of claims 1 to 12, wherein said compound is capable of triggering dose-dependent IL-18R downstream signaling on IL- 18 reporter cells.

14. The compound according to any one of claims 1 to 13, wherein said compound is capable of inducing IFN-y release by peripheral blood mononuclear cells (PBMCs) in the presence of low-dose IL-12 (10 ng / ml).

15. A pharmaceutical composition comprising the compound according to any one of claims 1 to 14.

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