CXCL8 inhibitors for treatment of ocular mucosal pemphigoids and / or oral mucosal pemphigoids

Abstract

The present invention relates to CXCL8 inhibitors useful for the prevention and / or treatment of ocular mucosal pemphigoids and / or oral mucosal pemphigoids.

Description

Technical Field

[0001] The present invention relates to a CXCL8 inhibitor for preventing and / or treating ocular mucous membrane pemphigoid (OcMMP) and / or oral mucous membrane pemphigoid. Background Art

[0002] Mucous membrane pemphigoid (MMP) is a systemic, cicatricial autoimmune disease that primarily affects the orificial mucous membranes (eg, conjunctiva, nasal cavity, oropharynx, and genitalia).

[0003] Approximately 75% of MMP patients develop antibodies against BP180 (type XVII collagen) and 25% develop antibodies against laminin 332. In less than 5% of MMP patients, antibodies against type VII collagen or α6β4 integrin are detected (Domloge-Hultsch, N., et al., J Clin Invest, 1992. 90(4): p. 1628-33; Oyama, N., et al., Br J Dermatol, 2006. 154(1): p. 90-8; Schmidt, E., et al., Br J Dermatol, 2001. 145(5): p. 778-83).

[0004] Ocular involvement occurs in approximately 70% of all MMP cases, and ocular MMP is the leading cause of cicatricial conjunctivitis in developed countries.

[0005] Linear immunoglobulin A disease, mucosal-predominant epidermolysis bullosa acquisita, and anti-laminin 332 / anti-integrin / anti-laminin 5 pemphigoid fall under the category of ocular MMPs (OcMMPs).

[0006] The progressive inflammatory and scarring nature of ocular MMPs leads to severe visual impairment in 30% of affected eyes and bilateral blindness in 20%.

[0007] Ocular MMPs are often associated with oral mucosal lesions including exfoliative gingivitis, vesicles, pseudomembrane-covered erosions, and ulcers.

[0008] In some cases of MMP, only the oral mucosa is involved.

[0009] The underlying pathophysiology of this disease is a type 2 hypersensitivity reaction to the basal epithelial membrane of the conjunctiva.

[0010] In particular, conjunctival involvement is critical, as autoantibody-induced inflammation can lead to conjunctival scarring that can progress even after the inflammatory process has ceased and cause visual impairment and blindness.

[0011] Early diagnosis and appropriate treatment are crucial to avoid inflammatory and infectious complications and possible vision loss.

[0012] Ocular MMP management aims to control immune-mediated inflammatory diseases and prevent fibrosis and disease progression.

[0013] Georgoudis, P. et al., "Ocular Mucous Membrane Pemphigoid: Current State of Pathophysiology, Diagnostics and Treatment", Ophthalmol. Ther. (2019) 8, 5–17, discloses a stepladder approach to select immunosuppressants based on disease severity (mild, moderate, severe) and gradually intensify treatment. The drugs used are dapsone, sulfapyridine, sulfasalazine, azathioprine (AZA), methotrexate (MTX), mycophenolate mofetil (MMF), cyclophosphamide, and short-term oral steroids.

[0014] CD20 monoclonal antibodies, TNFα inhibitors, and intravenous immunoglobulin (IVIg) are used to treat the disease in patients who do not respond to conventional immunosuppressants.

[0015] The mainstay of treatment is high-dose systemic corticosteroids, supplemented with potential corticosteroid-sparing agents such as azathioprine, mycophenolate mofetil, dapsone, antibiotics with anti-inflammatory activity (e.g., doxycycline), high-dose intravenous immunoglobulin, and the anti-CD20 antibody rituximab.

[0016] Current treatments are often ineffective and associated with serious adverse events.

[0017] Therefore, there remains a great need for effective and safe treatments.

[0018] Several possible mechanisms have been proposed to explain the development of antibody-mediated disease MMPs.

[0019] CXCL8 (interleukin-8, IL-8) is an endogenous chemokine produced by most nucleated cells such as fibroblasts, macrophages, endothelial cells and epithelial cells.

[0020] As reported, the biological activity of CXCL8 is mediated by the interaction of CXCL8 with CXCR1 and CXCR2 membrane receptors, which belong to a family of seven transmembrane receptors and are expressed on the surface of human neutrophils and a variety of T cells (L. Xu et al., J. Leukocyte Biol., 57, 335, 1995). Although CXCR1 activation is known to play a key role in CXCL8-mediated chemotaxis, it has been speculated that CXCR2 activation may also play a pathophysiological role in chronic inflammatory diseases.

[0021] Various CXCL8 inhibitors have been developed and are known to those skilled in the art.

[0022] WO2000 / 024710 discloses N-(2-aryl-propionyl)-sulfonamides having inhibitory activity on neutrophil chemotaxis and degranulation induced by interleukin-8, and their use in preventing and treating tissue damage caused by excessive recruitment of polymorphonuclear neutrophils (PMN) at inflammatory sites, in particular in the treatment of psoriasis, rheumatoid arthritis, ulcerative colitis, acute respiratory insufficiency, idiopathic fibrosis, and glomerulonephritis.

[0023] WO2005 / 090295 discloses (R)-2-[4-(trifluoromethanesulfonyloxy)phenyl]propionic acid derivatives as inhibitors of polymorphonuclear cell and monocyte chemotaxis, particularly in the treatment of neutrophil-dependent pathological conditions. The compounds are also disclosed for use in the treatment of psoriasis, ulcerative colitis, melanoma, angiogenesis, chronic obstructive pulmonary disease (COPD), bullous pemphigoid, rheumatoid arthritis, idiopathic fibrosis, glomerulonephritis, and in the prevention and treatment of injury caused by ischemia and reperfusion.

[0024] WO2010 / 031835 discloses 2-aryl-propionic acids and derivatives substituted at the 4-position with a 2-aminoheterocycle as potent inhibitors of CXCL8-induced chemotaxis, which can be used to prevent and treat tissue damage caused by excessive recruitment of polymorphonuclear neutrophils (PMNs) at inflammatory sites. The compounds are also disclosed for use in treating transient cerebral ischemia, injury caused by ischemia and reperfusion, bullous pemphigoid, rheumatoid arthritis, idiopathic fibrosis, and glomerulonephritis.

[0025] The present invention aims to provide effective treatments for MMPs, particularly ocular and oral MMPs. Summary of the Invention

[0026] The present invention relates to CXCL8 inhibitors for use in preventing and / or treating ocular mucosal pemphigoid (OcMMP) and / or oral mucosal pemphigoid in a subject.

[0027] The present invention also relates to a pharmaceutical composition comprising a CXCL8 inhibitor and at least one pharmaceutically acceptable excipient or carrier for preventing and / or treating ocular mucosal pemphigoid and / or oral mucosal pemphigoid in a subject.

[0028] The present invention also relates to a method for preventing and / or treating ocular mucosal pemphigoid and / or oral mucosal pemphigoid in a subject, comprising administering to a subject in need thereof an effective amount of one or more CXCL8 inhibitor compounds of the present invention.

[0029] The present invention also relates to the use of the claimed CXCL8 inhibitor in the preparation of a medicament for preventing and / or treating ocular mucosal pemphigoid and / or oral mucosal pemphigoid in a subject. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 The effect of treatment with DF2156A or reparixin by topical ocular administration or methylprednisolone (MP) by intraperitoneal injection is shown in an experimental MMP mouse model on day 12 after the initial anti-mLAMα3 IgG injection, compared to control animals treated with vehicle by topical ocular administration (vehicle). Animals treated (sc injection) with IgG isolated from normal rabbit serum (NR IgG) were used as negative controls. In detail:

[0031] Panel A shows the effect of treatment with DF2156A, reparixin, or methylprednisolone on the formation of palpebral conjunctival splits (expressed as conjunctival scores) at day 12.

[0032] Data are expressed as mean + / - standard deviation. At day 12, there was a statistically significant difference between the vehicle group and DF2156A (p = 0.0016). Except for NR IgG (n = 10), data are based on 12 to 14 mice per group. Asterisks indicate statistical significance (ANOVA with Holm Sidak method for multiple comparisons).

[0033] Panel B shows semi-quantification of subepithelial inflammatory infiltrates based on hematoxylin and eosin (H&E) stained biopsies of the palpebral conjunctiva in mice treated with DF2156A, Reparixin or methylprednisolone. On day 12, there was a statistically significant difference between the vehicle group and Reparixin (p=0.0474) and MP (p=0.0419). Animals treated (sc injection) with IgG isolated from normal rabbit serum (NR IgG) were used as negative controls. Data are expressed as mean + / - standard deviation. Except for NR IgG (n=3), data are based on 7 to 10 mice per group. Asterisks indicate statistical significance (ANOVA with Holm Sidak method for multiple comparisons).

[0034] Panel C shows the macroscopically affected ocular area evaluated at days 0, 4, 8, and 12 in mice treated with DF2156A or methylprednisolone (MP). The extent of the macroscopically affected ocular area is expressed as a percentage. Data are expressed as mean + / - standard deviation. On day 12, there were statistically significant differences between the vehicle group and DF2156A (p = 0.0165) and MP (p < 0.0001). Animals treated (sc injection) with IgG isolated from normal rabbit serum (NR IgG) were used as negative controls. Data are based on 12 to 14 mice per group (except NR IgG (n = 10)) and are expressed as mean + / - standard deviation. Mixed effects analysis with Dunnett's method for multiple comparisons.

[0035] Figure 2 Shown are representative clinical manifestations of the eyes and H&E-stained sections of the palpebral conjunctiva obtained 12 days after the initial anti-mLAMα3 IgG injection in mice treated with DF2156A or Reparixin by topical ocular administration or methylprednisolone (MP) by intraperitoneal injection.

[0036] Figure 3 Shown is the reduction of cleft formation in the palpebral conjunctiva in experimental MMP mice treated with DF2156A or Reparixin by topical ocular administration or with methylprednisolone (MP) by intraperitoneal injection, 28 days after the initial anti-mLAMα3 IgG injection. In detail:

[0037] Panel A shows the effect of treatment with DF2156A, reparixin, or methylprednisolone compared to vehicle on palpebral conjunctival cleft formation (expressed as conjunctival score) at day 28.

[0038] Animals treated (sc injection) with IgG isolated from normal rabbit serum (NR IgG) were used as negative controls. Data are presented as mean + / - standard deviation. At day 28, there were statistically significant differences between the vehicle group and DF2156A (p = 0.0229) and Reparixin (p = 0.0412). With the exception of NR IgG (n = 9), data are based on 11 to 14 mice per group. Asterisks indicate statistical significance (ANOVA with Holm-Sidak method for multiple comparisons).

[0039] Figure B shows a semi-quantitative analysis of subepithelial inflammatory infiltrates based on hematoxylin and eosin (H&E) stained biopsies of the palpebral conjunctiva in mice treated with DF2156A, Reparixin, or methylprednisolone (MP) compared to vehicle. Animals treated (sc injection) with IgG isolated from normal rabbit serum (NR IgG) were used as negative controls. Data are expressed as mean + / - standard deviation. On day 28, there was a statistically significant difference between the vehicle group and DF2156A (p = 0.0031). In addition, Reparixin significantly reduced inflammatory infiltrates (p = 0.0280) when compared to vehicle-treated mice only. Except for NRIgG (n = 5), data are based on 7 to 9 mice per group. Asterisks indicate statistical significance based on ANOVA with Dunnett's method for multiple comparisons, and open circles indicate statistical significance by ANOVA with the uncorrected Fishers LSD method (not corrected for multiple comparisons).

[0040] Panel C shows the extent of the affected eye area over 28 days in mice treated with MP or DF2156A compared to vehicle, expressed as the area under the curve (AUC). Animals treated (sc injection) with IgG isolated from normal rabbit serum (NRIgG) were used as negative controls. Data are expressed as mean + / - standard deviation, and except for NR IgG (n = 9), data are based on 11 to 14 mice per group. On day 28, there were statistically significant differences between the vehicle group and DF2156A (p = 0.0113) and MP (p < 0.0001). Asterisks indicate statistical significance (ANOVA with Dunnett's method for multiple comparisons).

[0041] Figure 4Shown are representative clinical manifestations of eyes and H&E-stained sections of palpebral conjunctiva obtained 28 days after initial anti-mLAMα3 IgG injection in mice treated with DF2156A or Reparixin by topical ocular administration or methylprednisolone (MP) by intraperitoneal injection.

[0042] Figure 5 The effects of DF2156A and MP treatment on the severity of oral lesions (expressed as oral scores) at day 28 are shown. Data are expressed as mean + / - standard deviation. For day 28, there was a statistically significant difference between the vehicle group and DF2156A (p = 0.0152). MP treatment significantly improved the oral score compared to the vehicle-treated group (p = 0.0136). Except for NR IgG (n = 9), the data are based on 11 to 14 mice per group. Diamonds represent statistical significance by ANOVA with uncorrected Fishers LSD method (not corrected for multiple comparisons). DETAILED DESCRIPTION

[0043] It has been unexpectedly discovered that CXCL8 inhibitors are effective in preventing and / or treating ocular mucosal pemphigoid (OcMMP) and oral mucosal pemphigoid.

[0044] Therefore, the present invention relates to CXCL8 inhibitors for use in preventing and / or treating ocular mucosal pemphigoid (OcMMP) and / or oral mucosal pemphigoid in a subject.

[0045] According to a preferred embodiment, the present invention relates to a CXCL8 inhibitor for use in preventing and / or treating ocular mucosal pemphigoid in a subject.

[0046] According to another preferred embodiment, the present invention relates to a CXCL8 inhibitor for use in preventing and / or treating oral mucosal pemphigoid in a subject.

[0047] According to another preferred embodiment, the present invention relates to a CXCL8 inhibitor for use in preventing and / or treating ocular mucosal pemphigoid and oral mucosal pemphigoid in a subject.

[0048] As used herein, the terms "treating" and "preventing" refer to eradication / amelioration of the disease being treated or one or more symptoms associated therewith, or prevention / delay of its onset, respectively, notwithstanding the fact that the patient may still have the underlying disease.

[0049] The term "CXCL8 inhibitor" according to the present invention means any compound that is capable of inhibiting the biological activity of CXCL8.

[0050] Methods for determining inhibition of CXCL8 biological activity and for classifying compounds as “CXCL8 inhibitors” are known in the art and are described, for example, in Moriconi et al., J. Med. Chem. 2007, 50, 3984-4002, and Brandolini et al., Scientific Reports (2019) 9: 11729.

[0051] Preferably, the CXCL8 inhibitor according to the present invention is a CXCL8 receptor inhibitor.

[0052] Preferably, the CXCL8 receptor inhibitor is a CXCR1 inhibitor or a CXCR1 / 2 inhibitor, which inhibits the activity of CXCL8 mediated by the CXCR1 receptor or by both the CXCR1 receptor and the CXCR2 receptor.

[0053] The CXCL8 receptor inhibitor preferably inhibits the binding of CXCL8 to the CXCR1 receptor (CXCR1 receptor inhibitor) or to both the CXCR1 receptor and the CXCR2 receptor (dual CXCR1 and CXCR2 receptor inhibitor), or prevents or blocks intracellular signaling activated by the binding of CXCL8 to the CXCR1 receptor (CXCR1 receptor inhibitor) or to both the CXCR1 receptor and the CXCR2 receptor (dual CXCR1 and CXCR2 receptor inhibitor).

[0054] According to a preferred embodiment, the CXCL8 receptor inhibitor is an antagonist of the CXCR1 receptor or an antagonist of both the CXCR1 receptor and the CXCR2 receptor.

[0055] According to another preferred embodiment, the CXCL8 receptor inhibitor is an allosteric inhibitor or orthosteric antagonist of the CXCR1 receptor or both the CXCR1 receptor and the CXCR2 receptor.

[0056] Alternatively, a CXCL8 receptor inhibitor preferentially binds to CXCL8, thereby preventing it from binding to its receptor.

[0057] The CXCL8 receptor inhibitor is preferably capable of inhibiting PMN chemotaxis induced by 1 nM CXCL8 in an in vitro assay at a concentration equal to or lower than 500 nM, preferably lower than 100 nM by at least 60%, preferably at least 70%, more preferably at least 80%, even more preferably at least 90%.

[0058] More preferably, the IC of the CXCL8 receptor inhibitor against the CXCR1 receptor according to the present invention is 50 The values are in the low nanomolar range, preferably below 10 nanomolar, more preferably between 0.02 and 5 nanomolar.

[0059] According to another preferred embodiment, the CXCL8 inhibitor is selected from the group consisting of small molecules, peptides and antibodies, more preferably it is a small molecule.

[0060] The term "small molecule" refers to an organic compound having a molecular weight of 900 Daltons or less.

[0061] CXCL8 inhibitors, in particular CXCL8 receptor inhibitors as defined above, are well known in the art.

[0062] To date, several CXCL8 inhibitors, such as small molecules, peptides and antibodies, have been disclosed, many of which are currently undergoing clinical trials or used for treatment (Jie Jack, Expert Opinion Ther. Patents, 2001, 11(12), Chao J. et al., Bioorganic & Medicinal Chemistry Letters 17, 2007, p.3778-3783, Busch-Petersen J. Current Topics in Medicinal Chemistry, 2006, 6, p.1345-135, Allegretti et al, Immunology Letters 2012, Vol. 145, p.68-78).

[0063] Preferably, the CXCL8 inhibitor according to the present invention is selected from the group comprising (or selected from) the following:

[0064] - Anti-CXCL-8 antibodies ABCream, BMS-986253, and ABX-IL-8;

[0065] -RP-72, PAC-G-31-P, SCH-N;

[0066] -Navarixin, which has the following formula:

[0067]

[0068] -SX-517, which has the following formula:

[0069]

[0070] -SX-576, which has the following formula:

[0071]

[0072] -SX-682, which has the following formula:

[0073]

[0074] - a compound having the formula:

[0075]

[0076] as well as

[0077] -5-[3-(2-fluorophenyl)ureido]-1-(2-hydroxypropyl)-1H-pyrazole-4-carboxylic acid ethyl ester

[0078] -5-[3-(3-fluorophenyl)ureido]-1-(2-hydroxypropyl)-1H-pyrazole-4-carboxylic acid ethyl ester

[0079] -3-[2-[1(R)-(4-bromofuran-2-yl)propylamino]-3,4-dioxo-1-cyclobutenylamino]-2-hydroxy-N,N-dimethylbenzamide

[0080] -3-[2-[1(R)-(4-chlorofuran-2-yl)propylamino]-3,4-dioxo-1-cyclobutenylamino]-2-hydroxy-N,N-dimethylbenzamide

[0081] -4-[1(R)-(N-isopropylcarbamoyl)ethyl]phenyl trifluoromethanesulfonate

[0082] -2-hydroxy-3-[4-[1(R)-(4-isopropylfuran-2-yl)propylamino]-1-oxo-1,2,5-thiadiazol-3-ylamino]-N,N-dimethylbenzamide

[0083] -3-(2-chlorophenylamino)-7-nitro-4H-1,2,4-benzothiadiazin-5-ol 1,1-dioxide

[0084] -1-[3-[4-[3-(4-fluorophenyl)iso oxazol-5-yl]phenoxy]propyl]-4-methylpiperazine

[0085] -N-(2-[(2,3-difluorobenzyl)sulfanyl]-6-[[(2R,3S)-3,4-dihydroxybutan-2-yl]oxy]pyrimidin-4-yl)azetidine-1-sulfonamide; and

[0086] - Compounds of formula (I) and formula (II) described below.

[0087] According to a preferred embodiment, the CXCL8 inhibitor has the general formula (I) or a pharmaceutically acceptable salt thereof:

[0088]

[0089] in

[0090] R1 Selected from linear or branched C1-C6 alkyl, benzoyl, phenoxy and trifluoromethanesulfonyloxy;

[0091] R 2 is selected from hydrogen and linear or branched C1-C3 alkyl; and

[0092] R 3 It is a linear or branched C1-C6 alkyl group or a trifluoromethyl group.

[0093] According to the present invention, "C1-C6 alkyl" means a linear or branched alkyl chain containing 1 to 6 carbon atoms.

[0094] R 1 Preferably selected from benzoyl, isobutyl and trifluoromethanesulfonyloxy. 1 Preferably, it is connected to the benzene ring at the 3-position or the 4-position. 1 It is 3-benzoyl, 4-isobutyl or 4-trifluoromethanesulfonyloxy.

[0095] R 2 Preferably it is selected from hydrogen or methyl.

[0096] R 3 Preferably, R is selected from a linear or branched C1-C6 alkyl group, more preferably selected from a linear or branched C1-C3 alkyl group. 3 It's methyl.

[0097] The chiral carbon of the compound of formula (I) is in the RS or R configuration, more preferably it is in the R configuration.

[0098] Particularly preferred compounds of formula (I) according to the present invention are selected from:

[0099] -2-(4-isobutylphenyl)propionylmethanesulfonamide, preferably R-(-)-2-(4-isobutylphenyl)propionylmethanesulfonamide (also known as Reparixin) and pharmaceutically acceptable salts thereof, preferably lysine salts thereof, and

[0100] -2-[(4-trifluoromethanesulfonyloxy)phenyl]-N-methanesulfonylpropionamide, preferably R(-)-2-[(4-trifluoromethanesulfonyloxy)phenyl]-N-methanesulfonylpropionamide and pharmaceutically acceptable salts thereof, especially the sodium salt thereof (also known as ladarixin or DF2156A).

[0101] Compounds of formula (I) are described in WO 2000 / 024710 A1 and WO 2005 / 090295 A2, which also disclose their synthesis.

[0102] According to a preferred embodiment, the CXCL8 inhibitor has the general formula (II) or a pharmaceutically acceptable salt thereof:

[0103]

[0104] in:

[0105] R1 is hydrogen or CH3;

[0106] X is OH;

[0107] R2 is hydrogen or a linear C1-C4 alkyl group,

[0108] Y is a heteroatom selected from S, O and N,

[0109] Z is selected from a linear or branched C1-C4 alkyl group, a linear or branched C1-C4 alkoxy group, a halogenated C1-C3 alkyl group and a halogenated C1-C3 alkoxy group.

[0110] Preferably, the chiral carbon of the compound of formula (II) is in the R or S configuration, more preferably it is in the S configuration.

[0111] A particularly preferred compound of formula (II) according to the present invention is 2-(4-{[4-(trifluoromethyl)-1,3-thiazol-2-yl]amino}phenyl)propanoic acid, preferably (2S)-2-(4-{[4-(trifluoromethyl)-1,3-thiazol-2-yl]amino}phenyl)propanoic acid or its sodium salt.

[0112] Another particularly preferred compound of formula (II) according to the present invention is 2-methyl-2-(4-{[4-(trifluoromethyl)-1,3-thiazol-2-yl]amino}phenyl)propanoic acid (DF2726Y) or a pharmaceutically acceptable salt thereof, preferably the sodium salt (DF2726A).

[0113] Compounds of formula (II) are described in WO 2010 / 031835 A2, which also discloses methods for their synthesis.

[0114] The preferred CXCL8 inhibitor according to the present invention is DF2156A or Reparixin, more preferably the CXCL8 inhibitor is DF2156A.

[0115] According to a preferred embodiment, the present invention relates to a CXCL8 inhibitor for preventing and / or treating ocular mucosal pemphigoid, wherein the CXCL8 inhibitor is selected from:

[0116] -2-(4-isobutylphenyl)propionylmethanesulfonamide, preferably R-(-)-2-(4-isobutylphenyl)propionylmethanesulfonamide (also known as Reparixin) and pharmaceutically acceptable salts thereof, preferably lysine salts thereof, and -2-[(4-trifluoromethanesulfonyloxy)phenyl]-N-methanesulfonylpropionamide, preferably R(-)-2-[(4-trifluoromethanesulfonyloxy)phenyl]-N-methanesulfonylpropionamide and pharmaceutically acceptable salts thereof, in particular sodium salts thereof (also known as ladarixin or DF2156A).

[0117] According to another preferred embodiment, the present invention relates to a CXCL8 inhibitor for preventing and / or treating oral mucosal pemphigoid, wherein the CXCL8 inhibitor is 2-[(4-trifluoromethanesulfonyloxy)phenyl]-N-methylsulfonylpropionamide, preferably R(-)-2-[(4-trifluoromethanesulfonyloxy)phenyl]-N-methylsulfonylpropionamide or a pharmaceutically acceptable salt thereof, in particular its sodium salt (also known as ladarixin or DF2156A).

[0118] Preferably, the CXCL8 inhibitor for use according to the present invention is topically administered to the surface of the subject's eye. Preferably, the CXCL8 inhibitor for use according to the present invention is formulated in the form of eye drops.

[0119] As will be discussed in the experimental section, the inventors have shown that topical administration of a CXCL8 inhibitor to the surface of the eye results in improvements in both ocular and oral MMPs.

[0120] The present invention also relates to a pharmaceutical composition comprising the CXCL8 inhibitor as described above and at least one pharmaceutically acceptable excipient or carrier, for preventing and / or treating ocular mucosal pemphigoid and / or oral mucosal pemphigoid in a subject.

[0121] Preferably, the pharmaceutical composition is an ophthalmic composition suitable for topical administration to the surface of the eye.

[0122] Therefore, the present invention also relates to ophthalmic compositions comprising a therapeutically effective amount of a CXCL8 inhibitor as described above and at least one ophthalmologically acceptable excipient or carrier.

[0123] According to a preferred embodiment, the present invention also relates to an ophthalmic composition comprising a therapeutically effective amount of a CXCL8 inhibitor as described above and at least one ophthalmologically acceptable excipient or carrier, for preventing and / or treating ocular mucosal pemphigoid and / or oral mucosal pemphigoid in a subject.

[0124] An "ophthalmically acceptable excipient" is an inert excipient that allows the drug to be delivered to the eye and / or eyelid to treat an ocular disease or disorder without causing deleterious effects to the eye.

[0125] According to one embodiment, the ophthalmic composition may be a liquid eye drop composition for topical administration to the anterior segment of the eye.

[0126] The liquid composition may be in the form of a solution, emulsion or suspension.The liquid composition may comprise micelles.

[0127] In one embodiment, the liquid composition is an aqueous composition.

[0128] Preferably, the liquid composition is an aqueous eye drop composition.

[0129] Preferably, the liquid composition comprises an ophthalmically acceptable excipient selected from the group consisting of an ophthalmically acceptable viscosity enhancer, a permeation enhancer, a buffer, an osmolarity regulator, a preservative, and a surfactant.

[0130] The viscosity enhancer has the function of increasing the viscosity of the composition and improving its retention in the conjunctival sac, and is preferably selected from cellulose derivatives, preferably hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylmethylcellulose, methylcellulose; polyvinylpyrrolidone and gelling agents, preferably gellan gum, xanthan gum and carbopol-974.

[0131] The penetration enhancer has the function of increasing the permeability of the drug across the ocular membrane and is preferably selected from cyclodextrins, chelating agents, crown ethers, bile acids and bile salts.

[0132] The buffer has the function of providing and maintaining the correct pH for the formulation to be compatible with use in the eye, preferably a pH of 6 to 8. The preferred buffer is a phosphate buffer, but other buffers capable of maintaining the pH within the desired range are also included, especially those suitable for ophthalmic use.

[0133] Osmolarity regulators are salts that make liquid compositions isotonic with ocular fluids. A preferred salt is sodium chloride (NaCl), but other biologically acceptable salts such as potassium chloride (KCl), calcium chloride (CaCl2), and magnesium chloride (MgCl2), and mixtures thereof, may also be used.

[0134] Preservatives inhibit microbial activity. Suitable preservatives include, for example, quaternary ammonium compounds such as benzalkonium chloride, cetyltrimethylammonium bromide, and cetylpyridinium chloride.

[0135] The surfactant has the function of stabilizing the composition and is preferably selected from polysorbates such as Tween 80, poloxamers (such as Pluronic F68) or proteins (such as serum albumin).

[0136] The liquid eye drop composition may be part of a kit comprising the composition, a container for holding the composition, and a drop dispenser.

[0137] According to the present invention, a "therapeutically effective amount" is an amount sufficient to achieve treatment or prevention of a disease. Determination of an effective amount is well within the capabilities of those skilled in the art based on the achievement of the desired effect. The effective amount will depend on factors including, but not limited to, the subject's weight and / or the extent of the disease or undesirable condition the subject suffers from.

[0138] The present invention also relates to the use of the above-mentioned CXCL8 inhibitor in the preparation of a medicament for preventing and / or treating ocular mucosal pemphigoid and / or oral mucosal pemphigoid.

[0139] The present invention also relates to a method for preventing and / or treating ocular mucosal pemphigoid and / or oral mucosal pemphigoid, comprising administering to a subject in need thereof an effective amount of one or more CXCL8 inhibitors of the present invention.

[0140] The present invention is further illustrated by the following examples.

[0141] Example

[0142] A study was designed to investigate and demonstrate the effects of pharmacological inhibition of CXCR1 / 2 by the compounds DF2156A and Reparixin in a mouse model of MMP.

[0143] Materials and methods

[0144] Test compound

[0145] -Ladarixin sodium salt (DF2156A) (Dompé S.pA)

[0146] -Reparixin lysine salt (Dompé S.pA)

[0147] -Vehicle (Solvent, Dompé S.pA)-(Positive control)

[0148] -Methylprednisolone (MP) (from UKSH, Lübeck - supplier Sanofi) - (reference treatment)

[0149] Animal Model - Mouse

[0150] Adult C57Bl / 6 (B6) mice (male and female) at least 6 weeks old were used. Animals were maintained on a 12-hour light-dark cycle in the animal facility of the University of Lübeck. Mice were kept under SPF conditions and had free access to acidified drinking water and a standard chow diet. The protocol was approved by the Animal Ethics Committee of the Ministry of Agriculture and the Environment of Schleswig-Holstein.

[0151] Production, Isolation, and Characterization of Anti-mLAMα3 IgG

[0152] To generate anti-mLAMα3 IgG, New Zealand white rabbits were subcutaneously immunized with 250 μg of an equimolar mixture of two purified recombinant proteins (aa1656-1985 and aa2756-3330 of the murine laminin α3 chain produced in Escherichia coli (E. coli) as disclosed in Heppe, EN, et al., J Invest Dermatol, 2017, 137, 1709-1718) suspended in complete Freund's adjuvant. Animals were boosted twice with the same protein preparation in incomplete Freund's adjuvant. Immune sera were obtained at regular intervals and characterized by IF microscopy on frozen sections of mouse skin. IgG from rabbits immunized with recombinant fragments of murine mLAMα3 and IgG from unimmunized rabbits were affinity purified using protein G agarose affinity chromatography (Amersham Biosciences, Heidelberg, Germany). The reactivity of the IgG fraction on mouse skin was analyzed by IF microscopy (Sitaru et al., J Immunol 2006, 177: 3461-8). In addition, each batch of anti-mLAMα3 IgG was characterized in vivo for its ability to induce experimental MMP in C57Bl / 6 (B6) mice (WP1.1). From this experiment, the amount of antibody that causes moderate experimental MMP was determined; that is, the amount required to induce moderate conjunctival involvement in the MMP model induced by antibody transfer (i.e., a score of 1 to 3 in up to 50% of the animals on day 12). For this purpose, the intensity of conjunctival separation was determined histologically. The affected body surface area should be 3% to 8% at the concentration used. The conjunctival score was set histologically after death and ranged from 0 to 4, depending on the intensity of conjunctival separation. The required amount was determined to be 5 mg / injected anti-mLAMα3 IgG.

[0153] Study Design

[0154] Experimental MMP induction by repeated injections of anti-mLAMα3 IgG and treatment regimen

[0155] To test the effect of 0.5% DF2156A and 0.5% Reparixin on the severity of conjunctival involvement in experimental MMP, the disease was induced by repeated subcutaneous (sc) injections of anti-mLAMα3 IgG (5 mg / mouse) every other day (0, 2, (...) and 10) into adult B6 mice, which were treated twice a day by eye drops throughout the experiment. Three drops of eye drops were given to each eye twice a day. Mice treated with vehicle by eye drops or methylprednisolone (MP) by ip injection were used as treatment controls. Mice injected sc with IgG isolated from normal rabbit serum (NR IgG) were used as negative controls.

[0156] Primary endpoint - evaluation of conjunctival lesions

[0157] The primary endpoint of the experiment was the extent of conjunctival lesions determined by lesion histopathology (H&E staining) on days 12 and 28 following an established scoring system (Heppe, EN, et al., J Invest Dermatol, 2017, 137, 1709-1718). Specifically, biopsies were taken from the palpebral conjunctiva on days 12 and 28 and embedded in paraffin. 4.5 μm thick sections were cut from three different depths of the biopsies in triplicate and H&E stained for further quantification. In the H&E stained histology, those with palpebral conjunctiva-epithelium greater than 1000 μm were used for quantification. For this score, the length of the cleft formation (cleft = separation of the epithelium from the underlying dermal structure) was measured. The length of the crack was scored from 0 to 4, with no crack = 0, less than 100 μm = 1, less than 200 μm = 2, less than 300 μm = 3, and greater than or equal to 300 μm = 4. Cracks occurring at the ends of the tissue were excluded as they were likely artifacts of the cutting process. The final score was determined by the longest crack among the 9 possible sections.

[0158] Endpoints were analyzed on day 12 in the following groups:

[0159] Normal rabbit IgG (n=10)

[0160] ■ Anti-mLAMα3 IgG + solvent (positive control) (n=14)

[0161] ■ Anti-mLAMα3 IgG+MP (reference treatment) (n=13)

[0162] ■Anti-mLAMα3IgG+DF2156A (n=13)

[0163] ■ Anti-mLAMα3 IgG + Reparixin (n=14)

[0164] Endpoints were analyzed on Day 28 in the following groups:

[0165] Normal rabbit IgG (n=9)

[0166] ■ Anti-mLAMα3 IgG + solvent (positive control) (n=11)

[0167] ■ Anti-mLAMα3 IgG+MP (reference treatment) (n=11)

[0168] ■Anti-mLAMα3IgG+DF2156A (n=14)

[0169] ■ Anti-mLAMα3 IgG + Reparixin (n=12)

[0170] The experiments were performed at two independent time points, on day 12 and day 28 respectively, and included 7 mice / group and 5 mice / normal rabbit IgG.

[0171] Severity of oral lesions

[0172] The extent of oral lesions was determined by endoscopic examination (Videomed, Munich, Germany) on day 28 using an established scoring system (Heppe, EN, et al., J Invest Dermatol, 2017, 137, 1709-1718). Specifically, if lesions / blistering / scabs / erosions were present in each oral quarter of the mouse, one point was assigned to each affected quarter. The quarters were defined as: left buccal mucosa, right buccal mucosa, hypopharynx, and tongue. The maximum score was 4.

[0173] Statistical analysis

[0174] Statistical analysis was performed using GraphPad Prism (version 8.4.3). ANOVA was used to compare treatment effects across multiple groups. Dunnett's or Holm-Sidaks' multiple comparisons were used, as appropriate, to isolate one or more groups that differed from the others.

[0175] result

[0176] 1. Topical administration of DF2156A and reparixin reduces cleft formation in the palpebral conjunctiva in experimental MMP

[0177] Injection of rabbit anti-mLAMα3 IgG into adult B6 mice resulted in the induction of experimental MMPs continuously for 4 to 8 days after the first IgG injection.

[0178] After 12 days of topical drug administration as eye drops twice daily, it was determined that conjunctival cleft formation was significantly reduced in the DF2156A-treated group compared to mice receiving vehicle ( Figure 1 This was also demonstrated by the reduction in subepithelial inflammatory cell infiltration as determined by semi-quantification of H&E-stained palpebral conjunctival biopsies obtained on day 12 from mice treated with DF2156A compared to mice receiving vehicle ( Figure 1 B).

[0179] The effects of topically administered DF2156A and systemically administered MP (ip, once daily) on the extent of the affected eye area were also measured in parallel. On day 12, a reduction in the affected eye area was observed for both treatments ( Figure 1 C). On day 12, no conjunctival lesions were observed in mice injected with NR-IgG compared with the other groups ( Figure 2 ).

[0180] When administered topically via eye drops, twice-daily treatment for 28 days resulted in a significant reduction in palpebral conjunctival cleft formation in mice receiving DF2156A or reparixin ( Figure 3 A). This was also demonstrated by the reduction of subepithelial inflammatory infiltrate in semiquantitative H&E-stained biopsies of the palpebral conjunctiva ( Figure 3 B). The effects of topically administered DF2156A and systemically administered MP (ip, once daily) on the extent of the affected ocular area were also measured in parallel. On day 28, a reduction in the area was observed for both treatments ( Figure 3 C). On day 28, mice that received only NR-IgG from healthy rabbits showed no conjunctival lesions compared to the other groups ( Figure 4 ).

[0181] in conclusion

[0182] The data obtained show that:

[0183] - DF2156A treatment resulted in improvements in conjunctival cleft formation that were already present on day 12 and persisted until day 28, as well as improvements in inflammatory infiltrates on day 28;

[0184] - Reparixin treatment reduced inflammatory infiltrates in the palpebral conjunctiva after 12 and 28 days and also reduced conjunctival scores after 28 days.

[0185] Based on the data obtained, twice-daily topical administration of both DF2156A and Reparixin produced an overall benefit in diseased mice, with Reparixin's therapeutic effect having a slower onset ( Figure 2 and 4 ).

[0186] 2. Topical application of DF2156A reduces oral lesions in experimental MMP

[0187] Treatment with DF2156A reduced the severity of oral involvement 28 days after eye drop application ( Figure 5 ), where there was a statistically significant difference between the vehicle group and DF2156A (p=0.0152).

Claims

1. A CXCL8 inhibitor for use in preventing and / or treating ocular mucosal pemphigoid and / or oral mucosal pemphigoid in a subject.

2. The CXCL8 inhibitor for use according to claim 1, wherein the CXCL8 inhibitor is topically administered to the surface of the eye of the subject.

3. The CXCL8 inhibitor for use according to claim 1 or claim 2, which is a CXCL8 receptor inhibitor selected from a CXCR1 inhibitor or a CXCR1 / 2 inhibitor. The CXCL8 inhibitor for use according to any one of claims 1 to 3, which is an antagonist of the CXCR1 receptor or an antagonist of both the CXCR1 receptor and the CXCR2 receptor. 5 . The CXCL8 inhibitor for use according to claim 1 , which is a CXCL8 receptor inhibitor selected from an allosteric inhibitor or an orthosteric antagonist of CXCR1 receptor or both CXCR1 receptor and CXCR2 receptor.

6. The CXCL8 inhibitor for use according to any one of claims 1 to 5, which has the general formula (I) or a pharmaceutically acceptable salt thereof: in R 1 Selected from linear or branched C1-C6 alkyl, benzoyl, phenoxy and trifluoromethanesulfonyloxy; R 2 is selected from hydrogen and linear or branched C1-C3 alkyl; and R 3 It is a linear or branched C1-C6 alkyl group or a trifluoromethyl group.

7. The CXCL8 inhibitor for use according to claim 6, wherein the chiral carbon of the compound of formula (I) is in the R configuration.

8. The CXCL8 inhibitor for use according to any one of claims 1 to 5, which has the general formula (II) or a pharmaceutically acceptable salt thereof: in: R1 is hydrogen or CH3; X is OH; R2 is hydrogen or a linear C1-C4 alkyl group, Y is a heteroatom selected from S, O and N, Z is selected from a linear or branched C1-C4 alkyl group, a linear or branched C1-C4 alkoxy group, a halogenated C1-C3 alkyl group and a halogenated C1-C3 alkoxy group.

9. The CXCL8 inhibitor for use according to claim 8, wherein the chiral carbon of the compound of formula (II) is in S configuration.

10. The CXCL8 inhibitor for use according to any one of claims 1 to 9, wherein the inhibitor is selected from: - Anti-CXCL8 antibodies ABCream, BMS-986253, and ABX-IL-8; -RP-72, PAC-G-31-P, SCH-N; -Navarixin, which has the following formula: -SX-517, which has the following formula: -SX-576, which has the following formula: -SX-682, which has the following formula: - a compound having the formula: as well as -5-[3-(2-fluorophenyl)ureido]-1-(2-hydroxypropyl)-1H-pyrazole-4-carboxylic acid ethyl ester -5-[3-(3-fluorophenyl)ureido]-1-(2-hydroxypropyl)-1H-pyrazole-4-carboxylic acid ethyl ester -3-[2-[1(R)-(4-bromofuran-2-yl)propylamino]-3,4-dioxo-1-cyclobutenylamino]-2-hydroxy-N,N-dimethylbenzamide -3-[2-[1(R)-(4-chlorofuran-2-yl)propylamino]-3,4-dioxo-1-cyclobutenylamino]-2-hydroxy-N,N-dimethylbenzamide -4-[1(R)-(N-isopropylcarbamoyl)ethyl]phenyl trifluoromethanesulfonate -2-hydroxy-3-[4-[1(R)-(4-isopropylfuran-2-yl)propylamino]-1-oxo-1,2,5-thiadiazol-3-ylamino]-N,N-dimethylbenzamide -3-(2-chlorophenylamino)-7-nitro-4H-1,2,4-benzothiadiazin-5-ol 1,1-dioxide -1-[3-[4-[3-(4-fluorophenyl)iso oxazol-5-yl]phenoxy]propyl]-4-methylpiperazine -N-(2-[(2,3-difluorobenzyl)sulfanyl]-6-[[(2R,3S)-3,4-dihydroxybutan-2-yl]oxy]pyrimidin-4-yl)azetidine-1-sulfonamide; and - Compounds of formula (I) and (II) as defined in claims 6 to 9.

11. The CXCL8 inhibitor for use according to claim 6 or 7, having a general formula (I) selected from: -2-(4-isobutylphenyl)propionylmethanesulfonamide, preferably R-(-)-2-(4-isobutylphenyl)propionylmethanesulfonamide and pharmaceutically acceptable salts thereof, preferably lysine salts thereof, and -2-[(4-trifluoromethanesulfonyloxy)phenyl]-N-methanesulfonylpropionamide, preferably R(-)-2-[(4-trifluoromethanesulfonyloxy)phenyl]-N-methanesulfonylpropionamide and its pharmaceutically acceptable salts, especially its sodium salt. 12 . The CXCL8 inhibitor for use according to claim 11 , which is the sodium salt of R(-)-2-[(4-trifluoromethanesulfonyloxy)phenyl]-N-methanesulfonylpropionamide.

13. The CXCL8 inhibitor for use according to claim 8 or 9, which has the general formula (II), which is 2-(4-{[4-(trifluoromethyl)-1,3-thiazol-2-yl]amino}phenyl)propanoic acid, preferably (2S)-2-(4-{[4-(trifluoromethyl)-1,3-thiazol-2-yl]amino}phenyl)propanoic acid or its sodium salt.

14. An ophthalmic composition comprising a therapeutically effective amount of a CXCL8 inhibitor as defined in claims 2 to 13 and at least one ophthalmologically acceptable excipient or carrier.

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