CXCL8 inhibitors for use in the treatment of ocular mucous membrane pemphigoid and / or oral mucous membrane pemphigoid

Abstract

The present invention relates to a CXCL8 inhibitor that is useful for the prevention and / or treatment of ocular mucous membrane pemphigoid and / or oral mucous membrane pemphigoid.

Description

[Technical Field]

[0001] The present invention relates to a CXCL8 inhibitor for use in the prevention and / or treatment of ocular mucous membrane pemphigoid (OcMMP) and / or oral mucous membrane pemphigoid. [Background technology]

[0002] Mucous membrane pemphigoid (MMP) is a systemic, scarring autoimmune disease that primarily affects the mucous membranes of orifices, such as the conjunctiva, nasal passages, oropharynx, and genitalia. Approximately 75% of MMP patients develop antibodies against BP180 (type XVII collagen), and 25% develop antibodies against laminin 332. Antibodies against type VII collagen or α6β4 integrin are detected in less than 5% of MMP patients (Domloge-Hultsch, N., et al., J Clin Invest, 1992.90(4):pp.1628-33; Oyama, N., et al., Br J Dermatol, 2006.154(1):pp.90-8; Schmidt, E., et al., Br J Dermatol, 2001.145(5):pp.778-83).

[0003] Ocular involvement occurs in approximately 70% of all MMP cases, and ocular MMP is the leading cause of cicatricial conjunctivitis in developed countries. Linear immunoglobulin A disease, mucosal-predominant epidermolysis bullosa acquisita, and antilaminin 332 / anti-epiligrin / anti-laminin 5 pemphigoid are included in the ocular MMPs (OcMMPs).

[0004] The progressive inflammation and scarring characteristic of ocular MMPs leads to severe visual impairment in 30% of affected eyes and bilateral blindness in 20%. Ocular MMP is often accompanied by oral mucosal lesions such as desquamative gingivitis, vesicles (small blisters), pseudomembrane-covered sores, and ulcers.

[0005] Some cases of MMP involve only the oral mucosa. The pathophysiological mechanism underlying this disease is a type 2 hypersensitivity reaction to the epidermal basement membrane of the conjunctiva.

[0006] Conjunctival involvement is particularly significant because autoantibody-induced inflammation leads to conjunctival scarring that can progress even after the inflammatory process has ceased, leading to visual impairment and blindness. Early diagnosis and appropriate treatment are crucial to avoid inflammatory and infectious complications and possible vision loss.

[0007] Management of ocular MMPs aims to control immune-mediated inflammatory disease and prevent fibrosis and disease progression. Georgoudis, P. et al. ["Ocular Mucous Membrane Pemphigoid: Current State of Pathophysiology, Diagnostics and Treatment," Ophthalmol. Ther. (2019) 8, 5-17] disclose that immunosuppressants are selected and treatment is intensified using a stepladder approach depending on the severity of the disease (mild, moderate, severe). The medications used are dapsone, sulfapyridine, sulfasalazine, azathioprine (AZA), methotrexate (MTX), mycophenolate mofetil (MMF), cyclophosphamide, and short-term oral steroids.

[0008] CD20 monoclonal antibodies, TNFα inhibitors, and intravenous immunoglobulin (IVIg) are used to treat disease in patients refractory to conventional immunosuppressants. The mainstay of treatment is high-dose systemic corticosteroids, supplemented with corticosteroid-sparing drugs such as azathioprine, mycophenolate, and dapsone, antibiotics with anti-inflammatory activity such as doxycycline, high-dose intravenous immunoglobulin, and the anti-CD20 antibody rituximab.

[0009] Current treatment regimes are often ineffective and associated with serious adverse events. Therefore, there remains a high need for effective and safe treatments. Several possible mechanisms underlying the development of the antibody-mediated disease MMP have been proposed.

[0010] CXCL8 (interleukin-8, IL-8) is an endogenous chemotactic factor produced by most nucleated cells, including fibroblasts, macrophages, endothelial cells, and epithelial cells.

[0011] It has been reported that the biological activity of CXCL8 is mediated by its interaction with the CXCR1 and CXCR2 membrane receptors, which belong to the seven-transmembrane receptor family and are expressed on the surface of human neutrophils and several types of T cells (L. Xu et al., J. Leukocyte Biol., 57, 335, 1995). CXCR1 activation is known to play an important role in CXCL8-mediated chemotaxis, but CXCR2 activation may also play a pathophysiological role in chronic inflammatory diseases.

[0012] A variety of CXCL8 inhibitors have been developed and are well known to those skilled in the art. WO2000 / 024710 discloses N-(2-aryl-propionyl)-sulfonamides having inhibitory activity against neutrophil chemotaxis and degranulation induced by interleukin-8, and their use in the prevention and treatment of tissue damage caused by the aggravated recruitment of polymorphonuclear neutrophils (white blood cells, PMN) at inflammatory sites, particularly in the treatment of psoriasis, rheumatoid arthritis, ulcerative colitis, acute respiratory failure, idiopathic fibrosis, and glomerulonephritis.

[0013] WO2005 / 090295 discloses (R)-2-[4-(trifluoromethanesulfonyloxy)phenyl]propionic acid derivatives as inhibitors of polymorphonuclear and monocyte chemotaxis, particularly for use in the treatment of neutrophil-dependent pathologies (lesions). The use of these compounds is also disclosed for 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 damage caused by ischemia and reperfusion.

[0014] WO2010 / 031835 discloses 2-arylpropionic acids and derivatives substituted with 2-aminoheterocycles at position 4 as effective inhibitors of CXCL8-induced chemotaxis, useful for preventing and treating tissue damage caused by increased recruitment of polymorphonuclear neutrophils (PMN leukocytes) at inflammatory sites. The use of the compounds in the treatment of transient cerebral ischemia, damage caused by ischemia and reperfusion, bullous pemphigoid, rheumatoid arthritis, idiopathic fibrosis, and glomerulonephritis is also disclosed.

[0015] The present invention aims to provide an effective treatment for MMPs, particularly ocular and oral MMPs. [Prior art documents] [Patent documents]

[0016] [Patent Document 1] International Patent Publication No. 2000 / 024710 [Patent Document 2] International Patent Publication No. 2005 / 090295 [Patent Document 3] International Patent Publication No. 2010 / 031835 [Non-patent literature]

[0017] [Non-Patent Document 1] Domloge-Hultsch, N., et al., J Clin Invest, 1992.90(4):p.1628-33 [Non-patent document 2] Oyama, N., et al.,Br J Dermatol,2006.154(1):p.90-8 [Non-patent document 3] Schmidt, E., et al.,Br J Dermatol,2001.145(5):p.778-83 [Non-patent document 4] Georgoudis,P.et al.,Ophthalmol.Ther.(2019)8,5-17 [Non-Patent Document 5] L. Xu et al., J. Leukocyte Biol., 57, 335, 1995 Summary of the Invention

[0018] The present invention is directed to a CXCL8 inhibitor for use in the prevention and / or treatment of ocular mucous membrane pemphigoid (OcMMP) and / or oral mucous membrane pemphigoid in a subject. The present invention is also directed to a pharmaceutical composition comprising a CXCL8 inhibitor and at least one pharmaceutically acceptable excipient or carrier for use in the prevention and / or treatment of ocular mucous membrane pemphigoid and / or oral mucous membrane pemphigoid in a subject.

[0019] The present invention is also directed to a method for preventing and / or treating ocular mucous membrane pemphigoid and / or oral mucous membrane pemphigoid in a subject, the method comprising administering to a subject in need thereof an effective amount of one or more CXCL8 inhibitor compounds of the present invention.

[0020] The present invention is also directed to the use of the claimed CXCL8 inhibitors in the manufacture of a medicament for the prevention and / or treatment of ocular mucous membrane pemphigoid and / or oral mucous membrane pemphigoid in a subject. [Brief explanation of the drawings]

[0021] [Figure 1] Figure 1 shows the effects of treatment with DF2156A or reparixin administered by topical ocular administration, or treatment with methylprednisolone (MP) administered intraperitoneally, in an experimental MMP mouse model 12 days after the first anti-mLAMα3 IgG injection, compared to vehicle-treated control animals administered by topical ocular administration (vehicle). Animals treated with subcutaneous injections of IgG isolated from normal rabbit serum (NR IgG) were used as negative controls. In detail: Panel A shows the effect of treatment with DF2156A, reparixin, or methylprednisolone on palpebral conjunctival fissure formation on day 12, as measured by conjunctival score.

[0022] Data are shown as mean ± standard deviation. On day 12, there was a statistically significant difference between the vehicle group and DF2156A (p = 0.0016). Data are based on 12–14 mice per group, except for NR IgG (n = 10). Asterisks indicate statistically significant differences (ANOVA with Holm-Sidaks method for multiple comparisons).

[0023] Panel B shows semiquantification 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, statistically significant differences were observed between the vehicle group and reparixin (p=0.0474) and MP (p=0.0419). Animals treated with subcutaneous injections of IgG isolated from normal rabbit serum (NR IgG) served as negative controls. Data are shown as mean ± standard deviation. Data are based on 7–10 mice per group, except for NR IgG (n=3). Asterisks indicate statistical significance (ANOVA with Holm-Sidaks method for multiple comparisons).

[0024] Panel C shows the gross ocular area assessed on days 0, 4, 8, and 12 in mice treated with DF2156A or methylprednisolone (MP). The extent of the gross ocular area is expressed as a percentage. Data are presented as mean ± standard deviation. On day 12, statistically significant differences were observed between the vehicle group and DF2156A (p = 0.0165) and MP (p < 0.0001). Animals treated with subcutaneous injections of IgG isolated from normal rabbit serum (NR IgG) were used as negative controls. Data are based on 12–14 mice per group, except for NR IgG (n = 10), and are presented as mean ± standard deviation. Mixed-effects analysis using Dunnett's method for multiple comparisons. [Figure 2] Figure 2 shows representative clinical images of eyes and H&E-stained sections of the palpebral conjunctiva obtained 12 days after the first anti-mLAMα3 IgG injection in mice treated with DF2156A or reparixin by topical ocular administration or methylprednisolone (MP) by intraperitoneal injection. [Figure 3] Figure 3 shows the reduction in palpebral conjunctival fissure formation 28 days after the first anti-mLAMα3 IgG injection in experimental MMP mice treated with DF2156A or reparixin by topical ocular administration, or methylprednisolone (MP) by intraperitoneal injection. In detail: Panel A shows the effect of treatment with DF2156A, reparixin, or methylprednisolone on palpebral conjunctival fissure formation on day 28, expressed as conjunctival score, compared to vehicle.

[0025] Animals treated with subcutaneous injections of IgG isolated from normal rabbit serum (NR IgG) served as negative controls. Data are shown as mean ± standard deviation. At day 28, statistically significant differences were observed between the vehicle group and DF2156A (p = 0.0229) and reparixin (p = 0.0412). Data are based on 11–14 mice per group, except for NR IgG (n = 9). Asterisks indicate statistical significance (ANOVA with Holm-Sidaks method for multiple comparisons).

[0026] Panel B shows semiquantification 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 with vehicle. Animals treated with subcutaneous injections of IgG isolated from normal rabbit serum (NR IgG) were used as negative controls. Data are shown as mean ± standard deviation. At day 28, there is a statistically significant difference between the vehicle group and DF2156A (p = 0.0031). Furthermore, reparixin significantly reduced inflammatory infiltrates when compared with vehicle-treated animals alone (p = 0.0280). Data are based on 7–9 mice per group, except for NR IgG (n = 5). Asterisks indicate statistically significant differences based on ANOVA with Dunnett's method for multiple comparisons, and open circles indicate statistically significant differences based on ANOVA with the uncorrected Fisher's LSD method (no correction for multiple comparisons).

[0027] Panel C shows the extent of the affected eye area, expressed as area under the curve (AUC), over 28 days in mice treated with MP or DF2156A compared with vehicle. Animals treated with subcutaneous injections of IgG isolated from normal rabbit serum (NR IgG) were used as negative controls. Data are presented as mean ± standard deviation and are based on 11–14 mice per group, except for NR IgG (n = 9). At day 28, statistically significant differences were observed between the vehicle group and DF2156A (p = 0.0113) and MP (p < 0.0001). Asterisks indicate statistically significant differences (ANOVA with Dunnett's method for multiple comparisons). [Figure 4] Figure 4 shows representative clinical images of eyes and H&E-stained sections of the palpebral conjunctiva obtained 28 days after the first anti-mLAMα3 IgG injection in mice treated with DF2156A or reparixin by topical ocular administration or methylprednisolone (MP) by intraperitoneal injection. [Figure 5] Figure 5 shows the effect of DF2156A and MP treatment on the severity of oral lesions, as measured by oral score, on day 28. Data are presented as mean ± standard deviation. On day 28, there was a statistically significant difference between the vehicle group and DF2156A (p = 0.0152). MP treatment significantly increased oral score compared to the vehicle-treated group (p = 0.0136). Data are based on 11–14 mice per group, except for NR IgG (n = 9). Diamonds indicate statistically significant differences by ANOVA with uncorrected Fisher's LSD (uncorrected for multiple comparisons). DETAILED DESCRIPTION OF THE INVENTION

[0028] Surprisingly, it has been found that CXCL8 inhibitors are effective in the prevention and / or treatment of ocular mucous membrane pemphigoid (OcMMP) and oral mucous membrane pemphigoid. Thus, the present invention is directed to a CXCL8 inhibitor for use in the prevention and / or treatment of ocular mucous membrane pemphigoid (OcMMP) and / or oral mucous membrane pemphigoid in a subject.

[0029] According to a preferred embodiment, the present invention is directed to a CXCL8 inhibitor for use in the prevention and / or treatment of ocular mucous membrane pemphigoid in a subject. According to a further preferred aspect, the present invention is directed to a CXCL8 inhibitor for use in the prevention and / or treatment of oral mucous membrane pemphigoid in a subject.

[0030] According to a further preferred aspect, the present invention is directed to a CXCL8 inhibitor for use in the prevention and / or treatment of ocular mucous membrane pemphigoid and oral mucous membrane pemphigoid in a subject. As used herein, the terms "treatment" and "prevention" refer to the eradication / amelioration, or prevention / delay of onset, respectively, of the disorder being treated or one or more of its associated symptoms, regardless of the fact that the patient may still be afflicted with the underlying disease.

[0031] The term "CXCL8 inhibitor" according to the present invention means any compound capable of inhibiting the biological activity of CXCL8. Methods for measuring inhibition of the biological activity of CXCL8 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.

[0032] Preferably, the CXCL8 inhibitor according to the present invention is a CXCL8 receptor inhibitor. 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 and CXCR2 receptors.

[0033] The CXCL8 receptor inhibitor preferably inhibits the binding of CXCL8 to the CXCR1 receptor (CXCR1 receptor inhibitor) or to both the CXCR1 and CXCR2 receptors (CXCR1 and CXCR2 receptor dual 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 and CXCR2 receptors (CXCR1 and CXCR2 receptor dual inhibitor).

[0034] In a preferred embodiment, the CXCL8 receptor inhibitor is an antagonist of the CXCR1 receptor or an antagonist of both the CXCR1 and CXCR2 receptors. In another preferred embodiment, the CXCL8 receptor inhibitor is an allosteric inhibitor or orthosteric antagonist of the CXCR1 receptor or of both the CXCR1 and CXCR2 receptors.

[0035] Alternatively, the CXCL8 receptor inhibitor preferably binds to CXCL8, thereby preventing it from binding to its receptor. The CXCL8 receptor inhibitor is preferably capable of inhibiting PMN chemotaxis induced by 1 nM CXCL8 in an in vitro assay by at least 60%, preferably at least 70%, more preferably at least 80%, and even more preferably at least 90% at a concentration of 500 nM or less, preferably less than 100 nM.

[0036] More preferably, the CXCL8 receptor inhibitor according to the present invention has an IC50 activity against the CXCR1 receptor in the low nanomolar range, preferably below 10 nanomolar, more preferably in the range of 0.02 to 5 nanomolar. 50 It has a value.

[0037] According to a further preferred embodiment, said CXCL8 inhibitor is selected from a small molecule, a peptide and an antibody, more preferably it is a small molecule. The term "small molecule" refers to an organic compound having a molecular weight below 900 daltons.

[0038] CXCL8 inhibitors, in particular CXCL8 receptor inhibitors, as defined above are well known in the art. To date, several types of CXCL8 inhibitors, including small molecules, peptides, and antibodies, have been disclosed, many of which are currently undergoing clinical trials or are being used therapeutically (Jie Jack, Expert Opinion Ther. Patents, 2001, 11(12); Chao J. et al., Bioorganic & Medicinal Chemistry Letters 17, 2007, pp. 3778-3783; Busch-Petersen J. Current Topics in Medicinal Chemistry, 2006, 6, pp. 1345-135; Allegretti et al., Immunology Letters 2012, Vol. 145, pp. 68-78).

[0039] Preferably, the CXCL8 inhibitor according to the present invention is selected from the group comprising (or consisting of): - anti-CXCL8 antibodies ABCream, BMS-986253, and ABX-IL-8; - RP-72, PAC-G-31-P, SCH-N, - Formula:

[0040] [ka]

[0041] Navarixin, which has - Formula:

[0042] [ka]

[0043] SX-517 with - Formula:

[0044] [ka]

[0045] SX-576 with - Formula:

[0046] [ka]

[0047] SX-682 with - Formula:

[0048] [ka]

[0049] or

[0050] [ka]

[0051] or

[0052] [ka]

[0053] A compound having and - 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 - Trifluoromethanesulfonic acid 4-[1(R)-(N-isopropylcarbamoyl)ethyl]phenyl ester - 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)isoxazol-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 shown below.

[0054] According to one preferred embodiment, the CXCL8 inhibitor has the general formula (I):

[0055] [ka]

[0056] [In the formula, R 1 is selected from linear or branched C1-C6 alkyl, benzoyl, phenoxy, and trifluoromethanesulfonyloxy; R 2 is selected from hydrogen and straight-chain or branched C1-C3 alkyl; and R 3 is a straight chain or branched C1-C6 alkyl or trifluoromethyl; or a pharmaceutically acceptable salt thereof.

[0057] According to the present invention, "C1-C6 alkyl" denotes a straight or branched alkyl chain containing from 1 to 6 carbon atoms. R 1 is preferably selected from benzoyl, isobutyl, and trifluoromethanesulfonyloxy. 1 is preferably attached to the phenyl ring at the 3 or 4 position. According to the most preferred embodiment, R 1 is 3-benzoyl, 4-isobutyl or 4-trifluoromethanesulfonyloxy.

[0058] R 2 is preferably selected from hydrogen or methyl. R 3 is preferably selected from linear or branched C1-C6 alkyl, more preferably linear or branched C1-C3 alkyl. 3 is methyl.

[0059] The chiral carbon in the compounds of formula (I) may be in the RS or R configuration, more preferably it is in the R configuration. Particularly preferred compounds of formula (I) according to the invention are - 2-(4-isobutylphenyl)propionylmethanesulfonamide, preferably R-(-)-2-(4-isobutylphenyl)propionylmethanesulfonamide (also known as reparixin) and pharmaceutically acceptable salts thereof, preferably the lysine salt thereof, and 2-[(4-trifluoromethanesulfonyloxy)phenyl]-N-methanesulfonylpropionamide, preferably R(−)-2-[(4-trifluoromethanesulfonyloxy)phenyl]-N-methanesulfonylpropionamide and its pharmaceutically acceptable salts, in particular its sodium salt (also known as ladarixin or DF2156A).

[0060] Compounds of formula (I) are described in WO2000 / 024710A1 and WO2005 / 090295A2, and methods for their synthesis are also disclosed. According to one preferred embodiment, the CXCL8 inhibitor has the general formula (II):

[0061] [ka]

[0062] [In the formula, R1 is hydrogen or CH3; X is OH; R2 is hydrogen or linear C1-C4 alkyl; Y is a heteroatom selected from S, O and N; Z is selected from linear or branched C1-C4 alkyl, linear or branched C1-C4 alkoxy, haloC1-C3 alkyl, and haloC1-C3 alkoxy. or a pharmaceutically acceptable salt thereof.

[0063] 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. 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.

[0064] Another particularly preferred compound of formula (II) according to the invention is 2-methyl-2-(4-{[4-(trifluoromethyl)-1,3-thiazol-2-yl]amino}phenyl)propanoic acid (DF2726Y) or a pharmaceutically acceptable salt thereof, in particular the sodium salt (DF2726A).

[0065] Compounds of formula (II) are described in WO2010 / 031835A2, which also discloses methods for their synthesis. A preferred CXCL8 inhibitor according to the present invention is DF2156A or reparixin, more preferably the CXCL8 inhibitor is DF2156A.

[0066] According to a preferred aspect, the present invention is directed to a CXCL8 inhibitor for use in the prevention and / or treatment of ocular mucous membrane pemphigoid, the CXCL8 inhibitor comprising: - 2-(4-isobutylphenyl)propionylmethanesulfonamide, preferably R-(-)-2-(4-isobutylphenyl)propionylmethanesulfonamide (also known as reparixin) and pharmaceutically acceptable salts thereof, preferably the lysine salt thereof, and 2-[(4-trifluoromethanesulfonyloxy)phenyl]-N-methanesulfonylpropionamide, preferably R(−)-2-[(4-trifluoromethanesulfonyloxy)phenyl]-N-methanesulfonylpropionamide and its pharmaceutically acceptable salts, in particular its sodium salt (also known as ladarixin or DF2156A).

[0067] According to a further preferred aspect, the present invention is directed to a CXCL8 inhibitor for use in the prevention and / or treatment of oral mucous membrane pemphigoid, wherein the CXCL8 inhibitor is 2-[(4-trifluoromethanesulfonyloxy)phenyl]-N-methanesulfonylpropionamide, preferably R(-)-2-[(4-trifluoromethanesulfonyloxy)phenyl]-N-methanesulfonylpropionamide or a pharmaceutically acceptable salt thereof, in particular the sodium salt thereof (also known as ladarixin or DF2156A).

[0068] Preferably, the CXCL8 inhibitor for use according to the present invention is administered topically to the ocular surface of a subject. Preferably, the CXCL8 inhibitor for use according to the present invention is formulated in the form of eye drops.

[0069] As discussed in the experimental section, the inventors have shown that topical application of a CXCL8 inhibitor to the ocular surface results in improvements in both ocular and oral MMPs. The present invention is also directed to a pharmaceutical composition comprising the above-mentioned CXCL8 inhibitor and at least one pharmaceutically acceptable excipient or carrier for use in the prevention and / or treatment of ocular mucous membrane pemphigoid and / or oral mucous membrane pemphigoid in a subject.

[0070] Preferably, the pharmaceutical composition is an ophthalmic composition suitable for topical application to the ocular surface. Accordingly, the present invention is further directed to an ophthalmic composition comprising a therapeutically effective amount of the above-described CXCL8 inhibitor and at least one ophthalmologically acceptable excipient or carrier.

[0071] According to a preferred embodiment, the present invention is further directed to an ophthalmic composition comprising a therapeutically effective amount of the above-mentioned CXCL8 inhibitor and at least one ophthalmologically acceptable excipient or carrier for use in the prevention and / or treatment of ocular mucous membrane pemphigoid and / or oral mucous membrane pemphigoid in a subject.

[0072] An "ophthalmologically acceptable excipient" is an inert excipient that allows for delivery of a medication to the eye and / or eyelid to treat an ocular disease or condition without adverse effects on the eye. According to one embodiment, the ophthalmic composition may be a liquid eye drop composition for topical administration to the anterior portion of the eye.

[0073] The liquid composition may be in the form of a solution, emulsion or suspension. The liquid composition may also comprise micelles. In one embodiment, the liquid composition is an aqueous composition.

[0074] Preferably, the liquid composition is an aqueous eye drop composition. Preferably, the liquid composition comprises an ophthalmologically acceptable excipient selected from an ophthalmologically acceptable viscosity enhancer, penetration enhancer, buffer, osmolality adjuster, preservative, and surfactant.

[0075] The viscosity enhancing agent has the function of increasing the viscosity of the composition to improve its retention in the conjunctival sac and is preferably selected from cellulose derivatives, preferably hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylmethylcellulose, methylcellulose; polyvinylpyrrolidone and gelling agents, preferably gellan, xanthan gum and carbopol-974.

[0076] The penetration enhancer has the function of promoting the permeation of the drug through the ocular membrane, and is preferably selected from cyclodextrin, chelating agent, crown ether, bile acid and bile salt. The buffering agent functions to provide and maintain the proper pH of the formulation for compatibility with ophthalmic use, preferably between pH 6 and 8. A preferred buffering agent is phosphate buffer, although other buffering agents capable of maintaining the pH within the desired range are also included, particularly buffers suitable for ophthalmic use.

[0077] The tonicity adjusting agent is a salt capable of making the liquid composition isotonic with ocular fluid. A preferred salt is sodium chloride (NaCl), although other biologically acceptable salts, such as potassium chloride (KCl), calcium chloride (CaCl), and magnesium chloride (MgCl), and mixtures thereof, can also be used.

[0078] Preservatives inhibit microbial activity. Suitable preservatives include, for example, quaternary ammonium compounds, such as benzalkonium chloride, cetyltrimethylammonium bromide, and cetylpyridinium chloride.

[0079] The surfactant has the function of stabilizing the composition and is preferably selected from polysorbates such as Tween 80, poloxamers such as Pluronics F68, or proteins such as serum albumin.

[0080] The liquid eye drop composition may be part of a kit that includes the composition, a container for containing the composition, and an eye drop dispenser. A "therapeutically effective amount" according to the present invention means an amount sufficient to achieve treatment or prevention of a disease. Determining an effective amount is well within the capabilities of one skilled in the art based on achieving the desired effect. The effective amount will vary depending on factors such as, but not limited to, the weight of the subject and / or the severity of the disease or undesired symptoms from which the subject is suffering.

[0081] The present invention also relates to the use of the above-mentioned CXCL8 inhibitor in the manufacture of a medicament for the prevention and / or treatment of ocular mucous membrane pemphigoid and / or oral mucous membrane pemphigoid. The present invention is also directed to a method for preventing and / or treating ocular mucous membrane pemphigoid and / or oral mucous membrane pemphigoid, which method comprises administering to a subject in need thereof an effective amount of one or more CXCL8 inhibitors of the present invention.

[0082] The present invention is further illustrated by the following examples. [Example]

[0083] A study was designed to investigate and demonstrate the pharmacological inhibitory effects of CXCR1 / 2 exerted by compounds DF2156A and reparixin in an MMP mouse model. Materials and Methods Test Compound - Ladarixin sodium salt (DF2156A) (Dompe SpA) - Reparixin Lysine Salt (Dompe SpA) - Vehicle (solvent, Dompe SpA) - (positive control) - Methylprednisolone (MP) (supplier UKSH, Lübeck - supplier Sanofi) - (reference treatment) Animal Model - Mouse Adult C57Bl / 6 (B6) mice (male and female) aged 6 weeks or older were used. Animals were housed in the animal facility of the University of Lübeck under a 12-h light-dark cycle. Mice were housed under specific-pathogen-free conditions and provided with acidified drinking water and standard chow ad libitum. The protocol was approved by the Animal Rights Committee of the Ministry of Agriculture and Environment (Schleswig-Holstein).

[0084] Generation, isolation, and characterization of anti-mLAMα3 IgG To generate anti-mLAMα3 IgG, New Zealand White rabbits were subcutaneously inoculated with 250 μg of an equimolar mixture of two purified recombinant proteins (aa1656-1985 and aa2756-3330 of mouse laminin alpha 3 chain produced in E. coli as described in Heppe, E.N., 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 collected periodically and characterized by IF microscopy on frozen sections of mouse skin. IgG from rabbits immunized with and unimmunized against the recombinant fragment of mouse mLAMα3 was purified by affinity chromatography using protein G Sepharose (Amersham Biosciences, Heidelberg, Germany). The reactivity of the IgG fraction was analyzed by IF microscopy in mouse skin (Sitaru et al., J Immunol 2006, 177:3461-8). Each batch of anti-mLAMα3 IgG was further characterized in vivo for its ability to induce experimental MMP in C57Bl / 6 (B6) mice (WP1.1). From this experiment, the antibody dose required to induce moderate experimental MMP was determined. That is, the dose required to induce moderate conjunctival lesions (i.e., a score of 1–3 in up to 50% of animals by day 12) in the antibody transfer-induced MMP model. To this end, the intensity of conjunctival separation was determined histologically. The concentration used should affect 3–8% of the body surface area. Conjunctival scores were determined histologically after death and ranged from 0–4 depending on the intensity of conjunctival separation. The required dose was determined to be 5 mg / injection of anti-mLAMα3 IgG.

[0085] Study design Induction of experimental MMP by repeated injections of anti-mLAMα3 IgG and treatment protocol To test the effects of 0.5% DF2156A and 0.5% reparixin on the severity of experimental MMP-induced conjunctival lesions, adult B6 mice were induced with repeated subcutaneous (SC) injections of anti-mLAMα3 IgG (5 mg / mouse) every other day (days 0, 2, 3, 4, and 10). Mice were treated with eye drops twice daily throughout the experiment. Three drops of eye drops were administered twice daily to each eye. Mice treated with vehicle by eye drops or methylprednisolone by intraperitoneal (ip) injection served as treatment controls. Mice subcutaneously injected with IgG isolated from normal rabbit serum (NR IgG) served as negative controls.

[0086] Primary endpoint - Assessment of conjunctival lesions The primary endpoint of this study was the extent of conjunctival lesions, determined by histopathology (H&E staining) of the lesions on days 12 and 28 according to 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. Triplicate 4.5 μm-thick sections were cut from the biopsies at three different depths and stained with H&E for further quantification. H&E-stained tissues containing more than 1000 μm of palpebral conjunctival epithelium were used for quantification. The length of the split formation (split = epithelial detachment from the underlying skin structures) was measured for this score. The length of the split was graded from 0 to 4. No cracks = 0, less than 100 μm = 1, less than 200 μm = 2, less than 300 μm = 3, greater than 300 μm = 4. Cracks occurring at the edges of the tissue were excluded as they were considered to be of an artificial nature due to the cutting. The longest crack out of nine possible sections determines the final score.

[0087] Endpoints were analyzed on day 12 in the following groups: Normal rabbit IgG (n=10) Anti-mLAMα3 IgG + vehicle (positive control) (n=14) ·Anti-mLAMα3 IgG + MP (reference treatment) (n=13) ·Anti-mLAMα3 IgG + DF2156A (n=13) Anti-mLAMα3 IgG + reparixin (n=14) Endpoints were analyzed at day 28 in the following groups: Normal rabbit IgG (n=9) Anti-mLAMα3 IgG + vehicle (positive control) (n=11) ·Anti-mLAMα3 IgG + MP (reference treatment) (n=11) ·Anti-mLAMα3 IgG + DF2156A (n=14) Anti-mLAMα3 IgG + reparixin (n=12) The experiment included 7 mice / group and 5 mice / normal rabbit IgG and was carried out at two independent time points, 12 and 28 days, respectively.

[0088] Severity of oral lesions The extent of oral lesions was determined by endoscopy (Videomed, Munich, Germany) on day 28 according to an established scoring system (Heppe, EN, et al., J Invest Dermatol, 2017, 137, 1709-1718). Specifically, if lesions / blisters / crusts / erosions were present, each of the affected oral quadrants of the mice was counted as 1 point. The quadrants were defined as the left buccal mucosa, right buccal mucosa, hypopharynx, and tongue. The maximum score was 4.

[0089] statistical analysis Statistical analysis was performed using GraphPad Prism (version 8.4.3). ANOVA was used to compare treatment effects across groups. Dunnett's or Holm Sidaks' multiple comparisons were used, where appropriate, to separate distinct group(s).

[0090] result 1. Topical application of DF2156A and reparixin reduced palpebral conjunctival fissure formation in experimental MMP Injection of rabbit anti-mLAMα3 IgG into adult B6 mice results in sustained induction of experimental MMP within 4–8 days after the first IgG injection.

[0091] After topical application of the drug twice daily as eye drops for 12 days, DF2156A treatment significantly reduced conjunctival fissure formation compared with vehicle-treated mice (Fig. 1A), as reflected by semiquantification of H&E-stained palpebral conjunctival biopsies taken on day 12 from DF2156A-treated mice, which showed reduced subepithelial inflammatory cell infiltration compared with biopsies from vehicle-treated mice (Fig. 1B).

[0092] The effect of topically administered DF2156A on the extent of the affected eye area was also measured in parallel with that of systemically administered MP (intraperitoneally once daily). Both treatments resulted in a reduction in the affected eye area on day 12 (Figure 1C). Mice injected with NR-IgG showed no conjunctival lesions on day 12 compared with the other groups (Figure 2).

[0093] Treatment with topical eye drops applied twice daily for 28 days resulted in a significant reduction in palpebral conjunctival fissure formation in mice receiving DF2156A or reparixin (Figure 3A). This was reflected by a reduction in subepithelial inflammatory infiltrates, as determined by semiquantification of H&E-stained biopsies of the palpebral conjunctiva (Figure 3B). The effect of topically administered DF2156A on the extent of the affected ocular area was also measured in parallel with that of systemically administered MP (intraperitoneally administered once daily). A reduction in the area was observed with both treatments at day 28 (Figure 3C). Mice receiving only NR-IgG from healthy rabbits showed no conjunctival lesions at day 28 compared to the other groups (Figure 4).

[0094] conclusion The data obtained showed the following: - DF2156A treatment resulted in an improvement in conjunctival fissure formation already from day 12 to day 28, and in inflammatory infiltrates at day 28. - Reparixin treatment reduced the inflammatory infiltrate of the palpebral conjunctiva after 12 and 28 days, and also reduced the conjunctival score after 28 days.

[0095] Based on the data obtained, both DF2156A and reparixin administered topically twice daily provided overall benefit to diseased mice, with reparixin having a slower onset of therapeutic effect (Figures 2 and 4). 2. Topical application of DF2156A reduces oral lesions in experimental MMP Treatment with DF2156A reduced the severity of oral lesions after 28 days of eye drop application, with a statistically significant difference between the vehicle group and DF2156A (p=0.0152) (FIG. 5).

Claims

1. A pharmaceutical composition comprising a CXCL8 inhibitor for use in the prevention and / or treatment of ocular mucosal pemphigoid and / or oral mucosal pemphigoid in a subject.

2. The pharmaceutical composition according to claim 1, which is administered topically to the surface of the eye of a target.

3. The pharmaceutical composition according to claim 1 or claim 2, wherein the CXCL8 inhibitor is a CXCL8 receptor inhibitor selected from a CXCR1 inhibitor or a CXCR1 / 2 inhibitor.

4. The pharmaceutical composition according to claim 1 or 2, wherein the CXCL8 inhibitor is an antagonist of the CXCR1 receptor or an antagonist of both the CXCR1 and CXCR2 receptors.

5. The pharmaceutical composition according to claim 3, wherein the CXCL8 receptor inhibitor is selected from allosteric inhibitors or orthosteric antagonists of the CXCR1 receptor or both the CXCR1 and CXCR2 receptors.

6. The CXCL8 inhibitor is a general formula (I): 【Chemistry 1】 [In the formula, R １ C is either linear or branched. １ -C ６ Selected from alkyl, benzoyl, phenoxy, and trifluoromethanesulfonyloxy; R ２ This includes hydrogen and linear or branched carbon atoms. １ -C ３ Selected from alkyl; and R ３ is a straight-chain or branched C １ -C ６ alkyl or trifluoromethyl], or a pharmaceutically acceptable salt thereof, the pharmaceutical composition according to claim 1 or claim 2.

7. The pharmaceutical composition according to claim 6, wherein the chiral carbon of the compound of formula (I) is in an R configuration.

8. The CXCL8 inhibitor is of general formula (II): 【Chemistry 2】 [In the formula, R1 is hydrogen or CH ３ And; X is OH; R2 is hydrogen or linear carbon. １ -C ４ It is alkyl, Y is a heteroatom selected from S, O, and N. Z is a linear or branched C １ -C ４ Alkyl, linear, or branched C １ -C ４ Alkoxy, Halo C １ -C ３ Alkyl and Halo C １ -C ３ The pharmaceutical composition according to claim 1 or claim 2, having [selected from alkoxys] or a pharmaceutically acceptable salt thereof.

9. The pharmaceutical composition according to claim 8, wherein the chiral carbon of the compound of formula (II) is in an S configuration.

10. The CXCL8 inhibitor mentioned above, - Anti-CXCL8 antibodies ABCream, BMS-986253, and ABX-IL-8; - RP-72, PAC-G-31-P, SCH-N, - Formula: 【Transformation 3】 Navalixin, which contains - Formula: 【Chemistry 4】 SX-517 - Formula: 【Transformation 5】 SX-576 - Formula: 【Transformation 6】 SX-682 - Formula: 【Transformation 7】 or 【Transformation 8】 or 【Chemistry 9】 Compounds containing and - 5-[3-(2-fluorophenyl)ureido]-1-(2-hydroxypropyl)-1H-pyrazole-4-carboxylate ethyl ester - 5-[3-(3-fluorophenyl)ureido]-1-(2-hydroxypropyl)-1H-pyrazole-4-carboxylate 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 - Trifluoromethanesulfonic acid 4-[1(R)-(N-isopropylcarbamoyl)ethyl]phenyl ester - 2-hydroxy-3-[4-[1(R)-(4-isopropylfuran-2-yl)propylamino]-1-oxo-1,2,5-thiadiazole-3-ylamino]-N,N-dimethylbenzamide - 3-(2-chlorophenylamino)-7-nitro-4H-1,2,4-benzothiadiazine-5-ol 1,1-dioxide - 1-[3-[4-[3-(4-fluorophenyl)isoxazole-5-yl]phenoxy]propyl]-4-methylpiperazine - N-(2-[(2,3-difluorobenzyl)sulfanyl]-6-[[(2R,3S)-3,4-dihydroxybutan-2-yl]oxy]pyrimidine-4-yl)azetidine-1-sulfonamide; and - Compound of formula (I) 【Chemistry 10】 [In the formula, R1 is selected from linear or branched C1-C6 alkyl, benzoyl, phenoxy, and trifluoromethanesulfonyloxy; R2 is selected from hydrogen and a linear or branched C1-C3 alkyl group; and R3 is a linear or branched C1-C6 alkyl or trifluoromethyl group, or a pharmaceutically acceptable salt thereof. and (II) compounds 【Chemistry 11】 [In the formula, 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 linear or branched C1-C4 alkyl, linear or branched C1-C4 alkoxy, halo C1-C3 alkyl and halo C1-C3 alkoxy, or a pharmaceutically acceptable salt thereof. A pharmaceutical composition according to claim 1 or claim 2, selected from the group consisting of the following.

11. A CXCL8 inhibitor having general formula (I) - 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 pharmaceutically acceptable salts thereof, particularly the sodium salt thereof. A pharmaceutical composition according to claim 6, selected from the above.

12. The pharmaceutical composition according to claim 11, wherein the CXCL8 inhibitor having general formula (I) is the sodium salt of R(-)-2-[(4-trifluoromethanesulfonyloxy)phenyl]-N-methanesulfonylpropionamide.

13. The pharmaceutical composition according to claim 8, wherein the CXCL8 inhibitor having general formula (II) is 2-(4-{[4-(trifluoromethyl)-1,3-thiazole-2-yl]amino}phenyl)propanoic acid, preferably (2S)-2-(4-{[4-(trifluoromethyl)-1,3-thiazole-2-yl]amino}phenyl)propanoic acid or its sodium salt.

14. A CXCR1 inhibitor or a CXCR1 / 2 inhibitor selected from: An ophthalmic composition comprising a therapeutically effective amount of a CXCL8 inhibitor and at least one ophthalmologically acceptable excipient or carrier.

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