Anethole trithione for the treatment of vasculitis

Abstract

The present invention relates to anethole trithione (ATT) for use in preventing, treating and / or reducing the severity or progression of vasculitis, particularly in Kawasaki Disease and Kawasaki Disease-like diseases, in a subject in need thereof.

Description

[Technical field]

[0001] The present invention relates to anethole trithione (ATT) for use in preventing, treating and / or reducing the severity or progression of vasculitis, particularly of Kawasaki Disease and Kawasaki Disease-like diseases such as multisystem inflammatory syndrome, in a subject in need thereof. [Background technology]

[0002] Systemic vasculitis is a heterogeneous group of diseases characterized by damage to the vessel wall that can result in occlusion and / or aneurysm formation.

[0003] According to the Chapel Hill Consensus Conference on the Nomenclature of Vasculitis (CHCC2012), vasculitides can be classified according to: a) Blood vessel size: Large vessel vasculitis (e.g. giant cell arteritis, Takayasu arteritis, etc.) Medium-sized vessel vasculitis (e.g. giant cell arteritis, Takayasu's arteritis, polyarteritis nodosa, Kawasaki disease, etc.) Small vessel vasculitis (e.g. antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis, microscopic polyangiitis, Wegener's disease, Churg-Strauss syndrome, antiglomerular basement membrane (GBM) disease, Henoch-Schönlein purpura, cryoglobulinemic vasculitis, hypocomplementemic urticarial vasculitis, thromboangiitis obliterans, etc.) Vasculitis affecting various blood vessels (Behçet's disease, Cogan's disease, etc.) b) Organ- or tissue-related: Single-organ vasculitis (e.g., cutaneous leukocytoclastic vasculitis, cutaneous arteritis, primary central nervous system vasculitis, focal aortitis, etc.) c) Systemic disease-related: Systemic disease-associated vasculitis (e.g. lupus vasculitis, rheumatoid vasculitis, sarcoid vasculitis, etc.) Vasculitis with a presumed etiology (e.g., hepatitis C virus-associated cryoglobulinemic vasculitis, hepatitis B virus-associated vasculitis, syphilis-associated aortitis, drug-associated immune complex vasculitis, drug-associated ANCA-associated vasculitis, cancer-associated vasculitis, etc.)

[0004] Nearly all primary vasculitides can target the heart. Even if cardiac manifestations are rarely the predominant one, they can be life-threatening and therefore require specific diagnostic and therapeutic strategies. This complication occurs more frequently in children (Weiss, 2012. Pediatr Clin North Am. 59(2):407-23), adults with systemic inflammatory conditions (Sy et al., 2016. Semin Arthritis Rheum. 45(4):475-82) or patients with cardiovascular comorbidities. Cardiac complications of vasculitides such as myocarditis, coronary arteritis, coronary artery disease, occurrence of ventricular arrhythmias and valvular disease can lead to congestive heart failure and represent poor prognostic factors requiring aggressive treatment.

[0005] Various pathogenic mechanisms have been implicated in the induction of vasculitis, including cell-mediated inflammation, immune complex-mediated inflammation, and autoantibody-mediated inflammation.

[0006] The contribution of innate immune cells, in particular macrophages, neutrophils, natural killer (NK) cells and γδ T cells, to the development of various types of vasculitis has been described in the art.

[0007] Macrophages in particular are resident phagocytes that play a key role in host defense as well as tissue repair and healing. Very recent evidence has placed macrophages at the center of steady-state tissue homeostasis dealing with waste and tissue regeneration. Macrophages are equipped with pattern recognition receptors (PRRs) that interact with pathogen-associated molecular patterns, enabling them to efficiently phagocytose pathogens and infected cells, and secrete defense-related mediators and inflammatory cytokines. Macrophages have an additional role as antigen-presenting cells, effectively bridging innate and adaptive immunity. In a feed-forward loop, activation of antigen-specific T cells results in the amplification of macrophage responses.

[0008] Macrophages are located in all organs to detect, ingest and process debris, dead cells and foreign bodies, and are abundant in chronically inflamed, non-healing lesions such as atherosclerotic plaques. Macrophages are now also recognized as central players in vasculitis, and in several vasculitic syndromes (giant cell arteritis, Takayasu's arteritis, Kawasaki disease, ANCA-associated vasculitis or primary central nervous system vasculitis, to name just a few), granuloma formation can occur due to highly activated tissue-destructive macrophages and surrounding T cells forming complex lymphoid fine structures.

[0009] It is well known that macrophages can survive for long periods in healthy tissues, and multiple subsets of tissue-resident macrophages have been identified, such as microglia, dermal macrophages, and splenic marginal zone and metallophilic macrophages. In contrast to such resident macrophages, vasculitic conditions can recruit circulating monocytes and allow them to develop into macrophages.

[0010] The current paradigm holds that macrophages differentiate from monocytes upon migration from the circulation into tissues. The process controlling monocyte entry into specialized tissue sites in the arterial wall is independent of the cell source and relies on the upregulation of molecules that mediate the arrest of circulating monocytes by the leukocyte adhesion cascade to activated endothelial cells (ECs).

[0011] The pathogenic role of macrophages in vasculitis lies in their ability to secrete soluble factors such as cytokines, chemokines, growth factors and enzymes. The secretion of these soluble factors may represent a major amplification system in vasculitis. Furthermore, macrophages form granulomatous infiltrates in the vessel wall, which leads to mural neovascularization, loss of medial smooth muscle cells, destruction of elastic membranous lamellae and elastin fibers in the medial layer, and neointima proliferation narrowing the lumen.

[0012] Kawasaki disease (KD), a vasculitic form of KD, was first described in Japan by Tomisaku Kawasaki in 1967 and is associated with cardiovascular symptoms and complications, especially left ventricular (LV) dysfunction, which occurs in 70%–50% of patients in the acute phase (Yu et al., 2010. Pediatr Cardiol. 31(6):807-12; Dionne et al., 2018. Int J Rheum Dis. 21(1):45-49), representing a major contributor to morbidity and mortality.

[0013] The annual incidence of hospitalizations for US patients with KD (19 / 100,000 children aged 5 years or younger) has not changed significantly over the last 20 years (Uehara & Belay, 2012. J Epidemiol. 22(2):79-85). Asian and black Americans are 2.5 and 1.5 times more likely to develop KD than whites, respectively, suggesting a genetic link (Uehara & Belay, 2012. J Epidemiol. 22(2):79-85; Onouchi, 2012. Circ J. 76(7):1581-6). Approximately 75%-80% of cases in the US occur in children aged 5 years or younger, with a median age at diagnosis of 1.5 years and a male:female ratio of approximately 1.5:1.4. Peak occurrence occurred between January and March, suggesting an environmental contribution as well (Uehara & Belay, 2012. J Epidemiol. 22(2):79-85; Burns et al., 2013. PLoS One. 8(9):e74529).

[0014] Although the overall inducer of KD remains unknown, infiltration of the coronary artery wall by immune cells has also been described (Noval Rivas &Arditi,2020.Nat Rev Rheumatol.16(7):391-405). Immunohistochemical analysis of human postmortem tissues in particular has shown that monocytes, macrophages and neutrophils accumulate in the arterial wall (Takahashi et al.,2018.Int J Rheum Dis.21(1):31-35). These cells synthesize and secrete various inflammatory cytokines and chemokines, including tumor necrosis factor (TNF)-α, interferon (IFN)-γ and interleukin-6 (IL-6), which activate endothelial cells and cause vasculitis syndromes.

[0015] Intravenous immunoglobulin (IVIg) and high-dose aspirin have traditionally been the cornerstone of KD management, although the role of aspirin has been questioned (Terai et al., 1997. J Pediatr. 131(6):888-93; Lee et al., 2013. Korean Circ J. 43(3):182-6). Corticosteroids have also been evaluated for the treatment of acute and refractory KD. A meta-analysis (n=1,011) found that the use of IVIg plus corticosteroids as initial treatment significantly reduced the risk of coronary artery abnormalities compared with IVIg alone (odds ratio [OR]=0.3; 95% [CI]:0.20-0.46) (Chen et al., 2013. Heart. 99(2):76-82). IVIg dose-dependently prevents the development of coronary artery aneurysms. Its mechanism of action is unknown, but the effect may be due to its modulation of cytokine production, effects on T cell activity, and inhibition of antibody synthesis (Newburger et al., 2016. J Am Coll Cardiol. 67(14):1738-49). However, coronary artery aneurysms continue to develop in 25%-29% of affected children and are the leading cause of acquired pediatric heart disease in developed countries (Dionne et al., 2019. Pediatrics. 143(6):e20183341).

[0016] Because available treatments such as IVIg in combination with corticosteroids only modestly reduce the occurrence of coronary artery aneurysms (Dionne et al., 2019. Pediatrics. 143(6):e20183341), there remains great interest in identifying new compounds that can reduce the occurrence of vasculitis and especially KD.

[0017] Here, the inventors surprisingly demonstrate that anethole trithione (also referred to as ATT, AOL or 5-(4-methoxyphenyl)-3H-1,2-dithiole-3-thione) is able (i) to reduce the contractility and maintain relaxation of arterial vessels with endothelial dysfunction (in other words, to prevent vasoconstriction of the vessels or to promote vasodilation of the vessels), and (ii) to inhibit the secretion of soluble factors, in particular TNF-α, by macrophages, thus paving the way for new therapies to prevent, treat and / or reduce the severity or progression of vasculitis regardless of vessel size. Summary of the Invention

[0018] The present invention relates to anethole trithione (ATT) or a structural analogue thereof for use in preventing, treating, and / or reducing the severity or progression of Kawasaki Disease or Kawasaki Disease-like illness in a subject in need thereof.

[0019] In one embodiment, preventing, treating, and / or reducing the severity or progression of Kawasaki Disease or Kawasaki Disease-like illness comprises reducing or inhibiting macrophage activation.

[0020] In one embodiment, reducing or inhibiting macrophage activation comprises decreasing the release of tumor necrosis factor (TNF)-α by said macrophages.

[0021] In one embodiment, preventing, treating, and / or reducing the severity or progression of Kawasaki Disease or Kawasaki Disease-like illness comprises reducing the contractility of arterial blood vessels having endothelial dysfunction, and / or preventing vasoconstriction of arterial blood vessels, and / or promoting vasodilation of arterial blood vessels.

[0022] In one embodiment, preventing, treating, and / or reducing the severity or progression of Kawasaki Disease or Kawasaki Disease-like illness comprises reducing the contractility of arterial blood vessels with impaired endothelial function.

[0023] In one embodiment, the disease is Kawasaki disease.

[0024] In one embodiment, the disease is a Kawasaki Disease-like disease. In one embodiment, the disease is a Kawasaki Disease-like disease associated with a viral infection. In one embodiment, the disease is a Kawasaki Disease-like disease associated with a viral infection selected from the group consisting of SARS-CoV-2 infection, cytomegalovirus infection, Epstein-Barr virus infection, and human immunodeficiency virus infection.

[0025] In one embodiment, the disease is Kawasaki-like illness in children with COVID-19.

[0026] In one embodiment, ATT or a structural analog thereof can be administered prior to, simultaneously with, or after an additional therapeutic agent.

[0027] In one embodiment, the additional therapeutic agent is selected from the group consisting of intravenous immunoglobulin (IVIg), aspirin, a corticosteroid, an immunosuppressant, and an alkylating agent.

[0028] In one embodiment, the additional therapeutic agent is IVIg.

[0029] In one embodiment, the subject is a child or teenager.

[0030] In one embodiment, the structural analog of ATT has formula (I): [ka] (In the formula, X represents S, O or NHOH, preferably X is S or O, more preferably X is S; Y represents CH, C or N, preferably Y is CH or N, more preferably Y is CH; R 1 , R 2 , R 4 and R 5 each independently represents hydrogen, hydroxy, halo, amino, alkylsulfonyl, aminosulfonyl, cyano, nitro, carboxy, aryl, alkoxy, haloalkyl, alkylamino, aminoalkyl, nitrooxyalkyl, or carboxyalkyl; R 3 is methoxy or hydroxy, or R 3 and R 2 together with the carbon atom to which they are attached form a five-membered heteroaryl moiety, where -R 3 -R 2 -A-CR 6 =B- or -B=CR 6 -A-, wherein A is O, S or NR 7 Here, R 7 represents hydrogen, C1-C8 alkyl or alkyloxycarbonyl; B represents CH or N, and R 6 represents hydrogen, hydroxy, halo, amino, alkylsulfonyl, aminosulfonyl, cyano, nitro, carboxy, aryl, alkoxy, haloalkyl, alkylamino, aminoalkyl, nitrooxyalkyl, or carboxyalkyl) It is a compound of the formula:

[0031] The present invention relates to anethole trithione (ATT) or a structural analogue thereof for use in preventing, treating, and / or reducing the severity or progression of vasculitis in a subject in need thereof, the method comprising the steps of: (i) reducing or inhibiting macrophage activation, in particular reducing the release of tumor necrosis factor (TNF)-α by said macrophages, and / or (ii) reducing the contractility of arterial vessels with endothelial dysfunction; and / or (iii) preventing vasoconstriction of arterial blood vessels; and / or (iv) promoting vasodilation of arterial blood vessels; and / or (v) Suppressing the contractility of arterial blood vessels with impaired endothelial function The present invention also relates to anethole trithione (ATT) or structural analogs thereof, including

[0032] In one embodiment, the vasculitis is selected from the group consisting of Kawasaki disease, Kawasaki disease-like disease, giant cell arteritis, Takayasu's arteritis, polyarteritis nodosa, antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis, microscopic polyangiitis, Wegener's disease, Churg-Strauss syndrome, antiglomerular basement membrane (GBM) disease, Henoch-Schönlein purpura, cryoglobulinemic vasculitis, hypocomplementemic urticarial vasculitis, thromboangiitis obliterans, Behcet's disease, Cogan's disease, cutaneous leukocytoclastic vasculitis, cutaneous arteritis, primary central nervous system vasculitis, focal aortitis, lupus vasculitis, rheumatoid vasculitis, sarcoid vasculitis, hepatitis C virus-associated cryoglobulinemic vasculitis, hepatitis B virus-associated vasculitis, syphilis-associated aortitis, drug-associated immune complex vasculitis, drug-associated ANCA-associated vasculitis, and cancer-associated vasculitis.

[0033] In one embodiment, the vasculitis is granulomatous vasculitis, hi one embodiment, the granulomatous vasculitis is selected from the group consisting of Kawasaki disease, Takayasu's arteritis, ANCA-associated vasculitis, Churg-Strauss syndrome, and primary central nervous system vasculitis.

[0034] The present invention also relates to anethole trithione (ATT) or a structural analogue thereof for use in preventing, treating, and / or reducing the severity or progression of a vasoconstrictive disorder in a subject in need thereof.

[0035] In one embodiment, the vasoconstrictive disorder is selected from the group consisting of coronary vasospasm, vasospastic angina, diabetic neuropathy, diabetic gangrene, vascular headache, cerebral vasospasm, reversible cerebral vasoconstriction syndrome (RCVS), Raynaud's disease, acrocyanosis, Buerger's disease, complex regional pain syndrome (CRPS), livedo reticularis, stroke, and post-traumatic dystrophy.

[0036] definition "Anethole trithione" or "ATT" or "AOL" or "5-(4-methoxyphenyl)-3H-1,2-dithiole-3-thione" all have the following formula: [ka] It refers to a substituted dithiolethione having the formula:

[0037] "Kawasaki disease", also known as "mucocutaneous lymph node syndrome", refers to a form of vasculitis that primarily affects children under the age of five (according to the NHS, 72% of children with Kawasaki disease in the UK between 1998 and 2003 were under the age of five). Typical symptoms include a high fever lasting five days or more, and possibly one or more of the following: a rash, swollen lymph nodes in the neck, dry red cracked lips, a swollen, bumpy red tongue (informally called "strawberry tongue"), redness in the mouth and back of the throat, swollen red hands and feet, and red eyes. Without proper early treatment, complications such as moderate inflammation of the blood vessels, especially the coronary arteries, can develop. These arterial inflammations can lead to cardiac complications and ultimately acquired heart disease, with death occurring in 2-3% of cases. Kawasaki disease can be subdivided into non-shock Kawasaki disease versus Kawasaki disease shock syndrome (KDSS) by specific diagnostic criteria of the presence of a systolic blood pressure consistently less than 20% of the mean systolic blood pressure of healthy individuals of the same age or signs of reduced perfusion in the peripheral circulation. The literature has also coined the terms "atypical Kawasaki disease," "Kawasaki disease-like illness," and most recently "multisystem inflammatory syndrome" or "MIS" to describe patients with coronary artery abnormalities who cannot meet the classical Kawasaki disease case definition but have compatible laboratory findings. Kawasaki disease-like illness has typically been observed in subjects suffering from viral infections such as infection with SARS-CoV-2, cytomegalovirus (CMV), Epstein-Barr virus (EBV), or human immunodeficiency virus (HIV).

[0038] "Reducing the severity or progression" and all conjugations thereof when referring to vasculitis refers to the partial alleviation, inhibition or amelioration of vasculitis and / or its associated symptoms or complications.

[0039] "Pharmaceutically acceptable excipient" refers to any inactive ingredient required for the formulation of active agents in a suitable dosage form without causing side effects, allergic reactions or other adverse reactions when administered to animals, preferably humans.It includes any and all solvents, diluents, carriers, fillers, bulking agents, binders, disintegrants, polymers, lubricants, glidants, surfactants, isotonicity agents, thickening or emulsifying agents, stabilizers, absorption enhancers, flavoring agents, preservatives, antioxidants, buffers or any combination thereof.For administration to humans, the formulation must meet the sterility, pyrogenicity, general safety and purity standards required by regulatory authorities, such as the FDA or EMA.

[0040] "Prevention" and all conjugations thereof when referring to vasculitis refers to the reduction or elimination of the occurrence of vasculitis and / or its associated symptoms or complications.

[0041] "Salt" of the compounds of the present invention is used herein to describe their acid addition and base addition salts. Suitable acid addition salts are formed from acids which form non-toxic salts. Non-limiting examples include acetate, trifluoroacetate, adipate, aspartate, benzoate, besylate, bicarbonate / carbonate, bisulfate / sulfate, borate, tetrafluoroborate, camsylate, citrate, cyclamate, edisylate, esylate, formate, fumarate, gluceptate, gluconate, glucuronate, hexafluorophosphate, hybenzate, hydrochloride / chloride, hydrobromide / bromide, iod ... The salts include hydrochloride / iodide, isethionate, lactate, malate, maleate, malonate, mesylate, methylsulfate, naphthylate, 2-napsylate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate / hydrogen phosphate / dihydrogen phosphate, pyroglutamate, saccharate, stearate, succinate, tannate, tartrate, tosylate, trifluoroacetate and xinofoate. Suitable base salts are formed from bases which form non-toxic salts. Non-limiting examples include aluminum, arginine, benzathine, calcium, choline, diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olamine, potassium, sodium, tromethamine, 2-(diethylamino)ethanol, ethanolamine, morpholine, 4-(2-hydroxyethyl)morpholine and zinc salts. Hemisalts of acids and bases may also be formed, such as hemisulfate and hemicalcium salts. Preferred pharma-ceutically acceptable salts include hydrochloride / chloride, hydrobromide / bromide, bisulfate / sulfate, nitrate, citrate and acetate salts.

[0042] "Solvate" is used herein to describe a compound in the present invention that contains stoichiometric or substoichiometric amounts of one or more pharma- ceutically acceptable solvent molecules, such as ethanol or water. The term "hydrate" refers to when said solvent is water.

[0043] "Subject" refers to an animal, including a human. In the sense of the present invention, a subject may be a "patient", i.e., a person undergoing medical care, undergoing or having undergone medical treatment, or being monitored for the development of a disease.

[0044] "Symptoms associated with vasculitis" and all its variants refer to clinical signs and characteristics that are suggestive of a diagnosis of vasculitis. These clinical signs and characteristics vary depending on which blood vessels and therefore which organ systems are affected. However, common symptoms experienced by many people with vasculitis include, but are not limited to, fever, fatigue, loss of appetite, unintentional weight loss, muscle and joint pain, and neurological problems such as numbness or weakness. Symptoms associated with some specific types of vasculitis include: Behcet's syndrome: mouth and genital ulcers, eye inflammation, acne-like skin lesions Churg-Strauss syndrome: Asthma Cryoglobulinemic vasculitis: Purpura in the lower extremities, arthritis, weakness, nerve damage (neuropathy) Giant cell arteritis: Headache, scalp tenderness, jaw pain when chewing, blurred or double vision, blindness Henoch-Schönlein purpura: abdominal pain, hematuria, joint pain, purpura on the buttocks, legs and feet Kawasaki disease: fever, skin rash, eye inflammation Microscopic polyangiitis: skin lesions, fever, unintentional weight loss, glomerulonephritis, nerve damage, Polyarteritis nodosa: Purpura, skin ulcers, muscle and joint pain, abdominal pain, renal dysfunction Takayasu's arteritis: Numbness or coldness in the extremities, slow or absent pulse, high blood pressure, headache, visual disturbances Thromboangiitis obliterans: pain in the hands, arms, feet and legs, ulcers on the fingers and toes Wegener's disease: stuffy nose, chronic sinus infections, nosebleeds

[0045] "Therapeutically effective amount" means a level or amount of an agent intended to (1) delay or prevent the onset of vasculitis, (2) slow or halt the progression, worsening or aggravation of one or more symptoms of vasculitis, (3) bring about remission of symptoms of vasculitis, (4) reduce the severity or incidence of vasculitis, or (5) cure vasculitis, without causing significant adverse or deleterious side effects on the target. A therapeutically effective amount may be administered prior to the onset of vasculitis for prophylactic treatment. Alternatively or additionally, a therapeutically effective amount may be administered after the onset of vasculitis for therapeutic treatment.

[0046] "Treatment" and all conjugations thereof when referring to vasculitis refers to the complete reduction, inhibition, delay in onset or cure of vasculitis and / or its associated symptoms or complications. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0047] The present invention relates to anethole trithione (ATT) for use in preventing, treating, and / or reducing the severity or progression of vasculitis in a subject in need thereof.The present invention also relates to a method for preventing, treating, and / or reducing the severity or progression of vasculitis in a subject in need thereof, comprising administering anethole trithione (ATT) to said subject.The present invention also relates to the use of anethole trithione (ATT) for the manufacture of a medicament for preventing, treating, and / or reducing the severity or progression of vasculitis in a subject in need thereof.

[0048] In the context of the present invention, the type of vasculitis to be prevented or treated includes, but is not limited to: Large vessel vasculitis (including but not limited to giant cell arteritis, Takayasu's arteritis), Medium-sized vasculitis (including but not limited to giant cell arteritis, Takayasu's arteritis, polyarteritis nodosa, and Kawasaki disease); Small vessel vasculitis (including but not limited to antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis, microscopic polyangiitis, Wegener's disease, Churg-Strauss syndrome, antiglomerular basement membrane (GBM) disease, Henoch-Schonlein purpura, cryoglobulinemic vasculitis, hypocomplementemic urticarial vasculitis, and thromboangiitis obliterans); Vasculitis affecting various blood vessels (including but not limited to Behçet's disease and Cogan's disease), Single-organ vasculitis (including but not limited to cutaneous leukocytoclastic vasculitis, cutaneous arteritis, primary central nervous system vasculitis, and focal aortitis), Systemic disease-associated vasculitis (including but not limited to lupus vasculitis, rheumatoid vasculitis, and sarcoid vasculitis); or Vasculitis with a presumed etiology (including but not limited to Hepatitis C virus-associated cryoglobulinemic vasculitis, Hepatitis B virus-associated vasculitis, syphilis-associated aortitis, drug-associated immune complex vasculitis, drug-associated ANCA-associated vasculitis, and cancer-associated vasculitis) It may be any type of vasculitis.

[0049] In one embodiment, the vasculitis is selected from the group consisting of large vessel vasculitis, medium-sized vessel vasculitis, small vessel vasculitis, vasculitis affecting multiple blood vessels, single organ vasculitis, systemic disease-associated vasculitis, and vasculitis with a putative etiology.

[0050] In one embodiment, the vasculitis comprises or is selected from the group consisting of Kawasaki disease, giant cell arteritis, Takayasu's arteritis, polyarteritis nodosa, ANCA-associated vasculitis, microscopic polyangiitis, Wegener's disease, Churg-Strauss syndrome, antiglomerular basement membrane (GBM) disease, Henoch-Schönlein purpura, cryoglobulinemic vasculitis, hypocomplementemic urticarial vasculitis, thromboangiitis obliterans, Behcet's disease, Cogan's disease, cutaneous leukocytoclastic vasculitis, cutaneous arteritis, primary central nervous system vasculitis, focal aortitis, lupus vasculitis, rheumatoid vasculitis, sarcoid vasculitis, hepatitis C virus-associated cryoglobulinemic vasculitis, hepatitis B virus-associated vasculitis, syphilis-associated aortitis, drug-associated immune complex vasculitis, drug-associated ANCA-associated vasculitis, and cancer-associated vasculitis.

[0051] In one embodiment, the vasculitis is selected from the group consisting of Kawasaki disease, Takayasu's arteritis, polyarteritis nodosa, ANCA-associated vasculitis, microscopic polyangiitis, Churg-Strauss syndrome, antiglomerular basement membrane (GBM) disease, Henoch-Schönlein purpura, cryoglobulinemic vasculitis, hypocomplementemic urticarial vasculitis, thromboangiitis obliterans, Cogan's disease, cutaneous leukocytoclastic vasculitis, cutaneous arteritis, primary central nervous system vasculitis, focal aortitis, lupus vasculitis, rheumatoid vasculitis, sarcoid vasculitis, hepatitis C virus-associated cryoglobulinemic vasculitis, hepatitis B virus-associated vasculitis, syphilis-associated aortitis, drug-associated immune complex vasculitis, drug-associated ANCA-associated vasculitis, and cancer-associated vasculitis.

[0052] In one embodiment, the vasculitis is selected from the group including or consisting of Kawasaki disease, ANCA-associated vasculitis, giant cell arteritis, Takayasu's arteritis, and Behcet's disease.

[0053] In one embodiment, the vasculitis is selected from the group including or consisting of Kawasaki disease, ANCA-associated vasculitis, and Takayasu's arteritis.

[0054] In one embodiment, the vasculitis is granulomatous vasculitis.

[0055] In one embodiment, the granulomatous vasculitis includes or is selected from the group consisting of Kawasaki disease, giant cell arteritis, Takayasu's arteritis, ANCA-associated vasculitis, Wegener's disease, Churg-Strauss syndrome, and primary central nervous system vasculitis.

[0056] In one embodiment, the granulomatous vasculitis is selected from the group consisting of Kawasaki disease, Takayasu's arteritis, ANCA-associated vasculitis, Churg-Strauss syndrome, and primary central nervous system vasculitis.

[0057] In one embodiment, the vasculitis is Kawasaki disease.

[0058] In one embodiment, the vasculitis is Kawasaki-like disease.

[0059] An example of a Kawasaki disease-like illness is Kawa-COVID-19. Kawa-COVID-19 (also known as “Childhood COVID-19-associated multisystem inflammatory syndrome” [MIS-C], “Child and adolescent multisystem inflammatory syndrome [MIS] temporally associated with COVID-19,” “Pediatric multisystem inflammatory syndrome [PIMS] temporally associated with SARS-CoV-2 infection” [PIMS-TS], “COVID-19-associated systemic inflammatory syndrome” [SISCoV] or “Kawasaki disease-like illness in children with COVID-19”) is a Kawasaki disease-like illness affecting young subjects (typically children and teenagers, particularly those aged 5 years and younger) infected with the SARS-CoV-2 virus, the cause of the 2019-2021 Covid-19 pandemic (Pouletty et al., 2020. Ann Rheum Dis. 79(8):999-1006; Dhar et al., 2021. Pediatr Res. Published online ahead of print).

[0060] In the context of the present invention, the term anethole trithione refers to 5-(4-methoxyphenyl)-3H-1,2-dithiole-3-thione, but is also intended to encompass pharma- ceutically acceptable salts or solvates thereof, as well as structural analogues such as the compounds of formula (I), (II) or (III) described in WO2018162581, which compounds are incorporated herein by reference.

[0061] In one embodiment, the structural analog of anethole trithione has formula (I): [ka] (In the formula, X represents S, O or NHOH, preferably X is S or O, more preferably X is S; Y represents CH, C or N, preferably Y is CH or N, more preferably Y is CH; R 1 , R 2 , R 4 and R 5 each independently represents hydrogen, hydroxy, halo, amino, alkylsulfonyl, aminosulfonyl, cyano, nitro, carboxy, aryl, alkoxy, haloalkyl, alkylamino, aminoalkyl, nitrooxyalkyl, or carboxyalkyl; R 3 is methoxy or hydroxy, or R 3 and R 2 together with the carbon atom to which they are attached form a five-membered heteroaryl moiety, where -R 3 -R 2 -A-CR 6 =B- or -B=CR 6 -A-, wherein A is O, S or NR 7 Here, R 7 represents hydrogen, C1-C8 alkyl or alkyloxycarbonyl; B represents CH or N, and R 6represents hydrogen, hydroxy, halo, amino, alkylsulfonyl, aminosulfonyl, cyano, nitro, carboxy, aryl, alkoxy, haloalkyl, alkylamino, aminoalkyl, nitrooxyalkyl, or carboxyalkyl) or a pharma- ceutically acceptable tautomer, salt or solvate thereof.

[0062] In one embodiment, the structural analogues of anethole trithione are 5-(4-hydroxyphenyl)-3H-1,2-dithiol-3-thione, 5-(4-hydroxyphenyl)-3H-1,2-dithiol-3-one, 5-(4-hydroxyphenyl)-3H-1,2-dithiol-3-one oxime, 5-(4-hydroxyphenyl)-3H-1,2,4-dithiazole-3-thione, 4-(4-hydroxyphenyl)-3H-1,2-dithiol-3-thione , 5-(2-hydroxybenzo[d]oxazol-5-yl)-3H-1,2-dithiol-3-thione, 5-(2-hydroxybenzo[d]thiazol-6-yl)-3H-1,2-dithiol-3-thione, 5-(benzofuran-5-yl)-3H-1,2-dithiol-3-thione and methyl 5-(3-thioxo-3H-1,2-dithiol-5-yl)-1H-indole-1-carboxylate.

[0063] In one embodiment, the structural analog of anethole trithione is 5-(4-hydroxyphenyl)-3H-1,2-dithiole-3-thione.

[0064] The present invention also encompasses a method for preventing, treating, and / or reducing the severity or progression of vasculitis in a subject in need thereof, comprising administering to said subject a composition comprising anethole trithione (ATT), and the use of such a composition comprising anethole trithione (ATT) for preventing, treating, and / or reducing the severity or progression of vasculitis in a subject in need thereof.

[0065] In one embodiment, the composition comprising anethole trithione (ATT) is a pharmaceutical composition and further comprises a pharma- ceutically acceptable excipient.

[0066] In one embodiment, the anethole trithione is formulated for administration to a subject in need thereof.

[0067] In one embodiment, anethole trithione or a composition comprising same may be administered systemically or locally.

[0068] In one embodiment, anethole trithione or a composition comprising same can be administered by injection, oral, topical, nasal, buccal, rectal, vaginal, intratracheal, endoscopic, transmucosal or transdermal administration.

[0069] In one embodiment, anethole trithione or a composition comprising same may be injected, preferably systemically injected.

[0070] Examples of systemic injections include, but are not limited to, intravenous (iv), subcutaneous (sq), intradermal (id), intramuscular (im), intra-arterial, intraparenteral, intranodal, intralymphatic, intraperitoneal (ip), intracranial, intracardiac, intralesional, intraprostatic, intravaginal, intrarectal, intrathecal, intranasal, intratumoral (it), intravesicular, and perfusion.

[0071] Examples of formulations of anethole trithione suitable for systemic injection include those with sulfobutylether-β-cyclodextrin, which are described in WO2020049166, incorporated herein by reference.

[0072] In one embodiment, anethole trithione or a composition comprising same may be administered orally.

[0073] Examples of formulations of anethole trithione suitable for oral administration include Felviten, Halpen, Hepasulfol, Heporal, Mucinol (Sanofi Aventis), Sialor (Paladin Laboratories, Pharmascience, Solvay, Zuoz Pharma), Sonicur (Solvay), Sulfarlem (Solvay, Agüettant, Edward Keller, Sanofi-Aventis), Sulfarlem S (EG Labo), Tiopropen, and Tiotrifar.

[0074] In one embodiment, anethole trithione or a composition comprising the same may be used with a delivery system that facilitates the delivery of anethole trithione to the central nervous system. For example, various blood-brain barrier (BBB) ​​permeability enhancers may be used to temporarily and reversibly increase the permeability of the blood-brain barrier to drugs. Such BBB permeability enhancers include, but are not limited to, leukotrienes, bradykinin agonists, histamine, tight junction disrupting agents (e.g., zonulin, zot), hyperosmotic solutions (e.g., mannitol), cytoskeleton contracting agents, and short-chain alkyl glycerols (e.g., 1-O-pentylglycerol). Oral, sublingual, parenteral, implant, nasal, and inhalation routes can provide delivery of anethole trithione to the central nervous system. In some embodiments, anethole trithione or a composition comprising the same can be administered to the central nervous system of a subject with minimal effects on the peripheral nervous system.

[0075] Of course, other suitable routes of administration are contemplated by the present invention, and the mode of administration will ultimately be determined by the attending physician within the scope of sound medical judgment. In addition to administration by injection (iv, ip, im, etc.), other routes such as aerosol or subcutaneous administration may be utilized.

[0076] The dosage administered to a subject is ultimately determined by the attending physician within the scope of sound medical judgment and is tailored to each subject individually.It should also be understood that the specific therapeutically effective amount for any particular subject depends on various factors, such as the specific condition being treated and the severity of said condition, the specific composition used, the subject's age, weight, general health, sex and diet, administration time, administration route, treatment duration, the drug used in combination or simultaneously with anethole trithione or the composition containing it, and similar factors well known in the medical field.

[0077] In one embodiment, anethole trithione can be administered prior to, simultaneously with, or after the additional therapeutic agent.

[0078] In one embodiment, anethole trithione can be administered as an add-on therapy after a further therapeutic agent.

[0079] As used herein, the term "add-on therapy" refers to a therapy given to support or enhance the therapeutic effect of a previous therapy (additional therapeutic agent) and / or to reduce side effects associated with the previous therapy (additional therapeutic agent).

[0080] Suitable additional therapeutic agents include, but are not limited to, intravenous immunoglobulin (IVIg), aspirin, corticosteroids, immunosuppressants, and alkylating agents.

[0081] As used herein, "intravenous immunoglobulin" or "IVIg" refers to immunoglobulin therapy using a mixture of "normal human immunoglobulins" (NHIG). IVIg is typically prepared from a pool of at least 1000 human plasma donations, which contains immunoglobulin G (IgG) antibodies against many of the viruses prevalent in the general population. IVIg is commercially available under several trade names, including Aragam® (Oxbridge), Flebogamma® DIF (Grifols), Gammagard S / D® (Baxter), Gammaplex® (BPL), Gamunex® (Grifols), Intratect® (Biotest UK), Kiovig® (Baxter), Octagam® (Octapharma), Privigen® (CSL Behring) and Vigam® (BPL).

[0082] Suitable examples of corticosteroids include those described in subgroup H02 of the Anatomical Therapeutic Classification of Antibodies.

[0083] Further suitable examples of corticosteroids include, but are not limited to, (i) For example, a. Natural glucocorticoids, including cortisone, cortodoxone, desoxycortone, hydrocortisone, prebediolone, and pregnenolone b. Chloroprednisone, cloprednol, difluprednate, fludrocortisone, flugestone acetate, fluocinolone, fluorometholone, fluperolone, fluprednisolone, loteprednol, medrysone, methylprednisolone, prednicarbate, prednisolone, prednisone, tixocortol, alclometasone, beclomethasone, betamethasone, clobetasol, clobetasone, clocortolone, cortivazol, desoximetasone, dexamethasone, diflorasone, diflucortolone, fluclorolone, flumethasone, fluocortol Synthetic glucocorticoids, including fluocortolone, fluprednidene, fluticasone, halometasone, meprednisone, mometasone, paramethasone, prednylidene, rimexolone, triamcinolone, urobetasol, amcinonide, budesonide, ciclesonide, deflazacort, desonide, fluclonide, fludroxycortide, flunisolide, fluocinolone acetonide, fluocinonide, formocortal, halcinonide, and triamcinolone acetonide. Glucocorticoids such as (ii) Mineralocorticoids, such as desoxycortone, hydrocortisone, fludrocortisone, methylprednisolone, prednisolone and prednisone. Examples include:

[0084] Suitable examples of immunosuppressants include those described in subgroup L04 of the Anatomical Therapeutic Chemical Classification.

[0085] Further suitable examples of immunosuppressants include, but are not limited to, the following: (i) For example, a. Antifolates, including aminopterin, methotrexate, pemetrexed, pralatrexate, pteropterin, raltitrexed, denopterin, trimetrexate, and pemetrexed; b. Purine analogs, including azathioprine, mycophenolic acid, mycophenolate mofetil, pentostatin, cladribine, clofarabine, fludarabine, nelarabine, thioguanine, and mercaptopurine, etc. c. Pyrimidine analogs, including leflunomide, teriflunomide, fluorouracil, capecitabine, doxifluridine, tegafur, tegafur / gimeracil / oteracil, carmofur, floxuridine, cytarabine, gemcitabine, azacitidine, and decitabine, and the like; D. Hydroxycarbamide Metabolic antagonists such as (ii) Macrolides such as, for example, tacrolimus, cyclosporine, pimecrolimus, abetimus and gusperimus. (iii) Immunomodulatory imide drugs, such as lenalidomide, pomalidomide, thalidomide and apremilast. (iv) IL-1 receptor antagonists, such as anakinra (v) mTOR inhibitors, such as, for example, sirolimus, everolimus, ridafololimus, temsirolimus, umirolimus, and zotarolimus. (vi) serum targeting antibodies such as, for example, anti-complement component 5 antibodies (e.g., eculizumab), anti-TNF antibodies (e.g., adalimumab, afelimomab, certolizumab pegol, etanercept, golimumab, infliximab, nerelimomab, pegsnercept), anti-interleukin 5 antibodies (e.g., mepolizumab), anti-immunoglobulin E antibodies (e.g., omalizumab), anti-interferon antibodies (e.g., faralimomab), anti-interleukin 6 antibodies (e.g., elsilimomab, filgotinib), anti-interleukin 12 / interleukin 23 antibodies (e.g., lebrikizumab, ustekinumab) and anti-interleukin 17A antibodies (e.g., secukinumab). (vii) For example, anti-CD3 antibodies (e.g., muromonab-CD3, otelixizumab, teplizumab, visilizumab), anti-CD4 antibodies (e.g., clenoliximab, keliximab, zanolimumab), anti-CD11a antibodies (e.g., efalizumab), anti-CD18 antibodies (e.g., erlizumab), anti-CD20 antibodies (e.g., obinutuzumab, rituximab, ocrelizumab, pascolizumab), anti-CD23 antibodies (e.g., gomiliximab, lumiliximab), anti-CD40 antibodies (e.g., teneliximab, toralizumab), anti-CD62L antibodies (e.g., acelizumab, Cell-targeting antibodies such as anti-CD80 antibodies (e.g., galiximab), anti-CD147 antibodies (e.g., gavilimomab), anti-CD154 antibodies (e.g., ruplizumab), anti-BLysS antibodies (e.g., belimumab, blisibimod), anti-CTLA-4 antibodies (e.g., abatacept, belatacept), anti-interleukin 2 receptor antibodies (e.g., basiliximab, daclizumab, inolimomab), anti-interleukin 6 receptor antibodies (e.g., tocilizumab) and anti-integrin antibodies (e.g., natalizumab, vedolizumab).

[0086] Suitable examples of alkylating agents include, but are not limited to, nitrogen mustards (e.g., chlormethine, cyclophosphamide, ifosfamide, trofosfamide, chlorambucil, melphalan, prednimustine, bendamustine, and uramustine), nitrosoureas (e.g., carmustine, lomustine, semustine, fotemustine, nimustine, ranimustine, and streptozocin), alkylsulfonates (e.g., busulfan, mannosulfan, and the like), and the like. and treosulfan), aziridines (such as carboquone, thiotepa, triaziquone, triethylenemelamine, benzodopa, meturedopa, and uredopa), hydrazines (such as procarbazine), triazenes (such as dacarbazine and temozolomide), altretamine, mitobronitol, pipobroman, actinomycin, bleomycin, mitomycin, and plicamycin.

[0087] In one embodiment, anethole trithione can be administered before, simultaneously with, or after intravenous immunoglobulin (IVIg).

[0088] In one embodiment, the subject is an animal, preferably a mammal, more preferably a primate, and even more preferably a human.

[0089] In one embodiment, the subject is male. In one embodiment, the subject is female.

[0090] In one embodiment, the subject is a child, i.e., a young human below the age of puberty or below the age of adulthood. In one embodiment, the child is a newborn, i.e., a young human less than 28 days old. In one embodiment, the child is an infant, i.e., a young human between 1 and 12 months of age. In one embodiment, the child is a toddler, i.e., a young human between 12 and 36 months of age. In one embodiment, the subject is a teenager, i.e., a young human between the ages of puberty and adulthood, such as 18, 19, 20, or 21 years of age.

[0091] In one embodiment, the subject is a child up to 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months of age. In one embodiment, the subject is a child 2, 3, 4, 5, 6, 7, 8, 9 or 10 years of age.

[0092] In one embodiment, the subject is a child between the ages of 3 and 12 months, hi one embodiment, the subject is a child up to the age of 5 years.

[0093] In one embodiment, the subject is an adult, ie, a human over the age of adulthood (eg, over 18, 19, 20, or 21 years of age), or alternatively, a human over the age of puberty.

[0094] In one embodiment, the subject is / has been diagnosed with vasculitis. In one embodiment, the subject is / has been diagnosed with Kawasaki Disease. In one embodiment, the subject is / has been diagnosed with Kawa-COVID-19.

[0095] The present invention also relates to a method for reducing the contractility of arterial blood vessels having endothelial dysfunction and maintaining relaxation of arterial blood vessels in a subject in need thereof, the method comprising administering anethole trithione (ATT) or a composition comprising same to the subject, and the use of anethole trithione (ATT) or a composition comprising same for reducing the contractility of arterial blood vessels having endothelial dysfunction and maintaining relaxation of arterial blood vessels.

[0096] "Maintaining arterial blood vessel relaxation" means preventing vasoconstriction of the arterial blood vessels and / or promoting vasodilation of the arterial blood vessels.

[0097] In one embodiment, the endothelial dysfunction is a decrease in endothelial function of arterial blood vessels.

[0098] The present invention therefore also has application in the prevention, treatment and / or reducing the severity or progression of diseases associated with or characterised by hypercontractility of arterial vessels with endothelial dysfunction, in particular arterial vessels with reduced endothelial function, and / or diseases associated with or characterised by vasoconstriction of arterial vessels.

[0099] Such disorders, also referred to as "vasoconstrictive disorders" or "vasospastic diseases", include, but are not limited to, coronary vasospasm, vasospastic angina, heart attack, diabetic neuropathy, diabetic gangrene, vascular headache, cerebral vasospasm, reversible cerebral vasoconstriction syndrome (RCVS), Raynaud's disease, acrocyanosis, Buerger's disease, complex regional pain syndrome (CRPS), livedo reticularis, stroke, and post-traumatic dystrophy.

[0100] The present invention also relates to a method for inhibiting the secretion of soluble factors, particularly TNF-α, by macrophages using anethole trithione (ATT), and to the use of anethole trithione (ATT) for inhibiting the secretion of soluble factors, particularly TNF-α, by macrophages.

[0101] Thus, the present invention also has application in the prevention, treatment and / or reduction of the severity or progression of diseases associated with or characterised by the secretion of soluble factors, particularly TNF-α, by macrophages. [Brief description of the drawings]

[0102] [Figure 1]Figure 1A-1C are a set of three graphs showing the effect of ATT on cytokine production by macrophages. Figure 1A shows ATT inhibition of TNF-α production in LPS-activated macrophages. Dunnett's multiple comparison test: **p<0.01; ***p<0.001. Figure 1B shows the reduction in production of TNF-α by LPS-activated human macrophages cultured with ATT. Dunnett's multiple comparison test: *p<0.05; **p<0.01. Figure 1C shows the reduction in production of IL-10 by LPS-activated human macrophages cultured with ATT. Dunnett's multiple comparison test: **p<0.01; ****p<0.0001. [Diagram 2] Figures 2A-B are a set of two graphs showing the effect of ATT on 5-HT reactivity of pulmonary artery rings in the presence of endothelial dysfunction mediated by CHAPS or L-NAME. Figure 2A shows 5-HT reactivity in the presence of endothelial dysfunction mediated by CHAPS. Dunnett's multiple comparison test: **p<0.01; ns: not significant. Figure 2B shows 5-HT reactivity in the presence of endothelial dysfunction mediated by L-NAME. Dunnett's multiple comparison test: *p<0.05; **p<0.01; ns: not significant. [Diagram 3] Graph showing the relaxant effect of ATT on phenylephrine-induced contraction of pulmonary artery rings in the presence of endothelial dysfunction mediated by CHAPS or L-NAME. Tension % is normalized to control tension % for each group (ATT vs. control, L-NAME+ATT vs. L-NAME, CHAPS+ATT vs. CHAPS). Dunnett's multiple comparison test: *p<0.05; **p<0.01; ns: not significant. [Figure 4] FIG. 1 is a graph showing the effect of acute or prophylactic treatment with ATT on 5-HT responsiveness of pulmonary artery rings from rats with PH. Data are expressed as percentage of control contractions. Statistical analysis: two-way ANOVA and Bonferroni post-hoc test. *p<0.05; **p<0.01; ***p<0.001; ns: not significant. [Diagram 5]FIG. 1 is a scheme of the in vivo protocol used to evaluate the effect of ATT in the LCWE-induced Kawasaki disease mouse model. [Figure 6] Figures 6A-6F are a set of six graphs showing the results of a human primary cardiomyocyte-based assay after 12 hours of exposure to serum from Kawasaki disease patients [KD serum], serum from Kawasaki disease patients with 1 μM ATT added [KD serum + ATT 1 μM], serum from Wegener's disease patients [WD serum], or serum from Kawasaki disease patients in the convalescent phase, i.e., at least one year after disease remission [convalescent serum]. Figure 6A shows the peak height (in μm) under four conditions. Figure 6B shows the % sarcomere shortening under four conditions. Figure 6C shows the maximum sarcomere departure velocity (in μm / sec) under four conditions. Figure 6D shows the maximum sarcomere return velocity (in μm / sec) under four conditions. Figure 6E shows the decay time (in seconds) to 90% relaxation under four conditions. Figure 6F shows the decay time (in seconds) to 30% relaxation under four conditions. *p<0.05; **p<0.01 (one-way ANOVA). EXAMPLES

[0103] The present invention is further illustrated by the following examples.

[0104] Example 1: Evaluation of ATT on cytokine production by macrophages material and method CD14 isolated from healthy donors + Human macrophages were cultured with LPS (1 μg / mL) and ATT (10 μM) for 48 h. At the end of the culture, supernatants were collected and cytokine production was assessed.

[0105] TNF-α production was measured by ELISA. TNF-α and IL-10 production was measured using multiplex detection techniques.

[0106] result Vasculitis is characterized by activation of macrophages and lymphocytes that leads to the release of inflammatory cytokines. Because macrophage activation results in the release of inflammatory cytokines, the aim of this study was to evaluate the effect of ATT (10 μM) on its ability to inhibit the production of these cytokines.

[0107] As shown in Figure 1A, ATT significantly reduced TNF-α production in LPS-activated human macrophages (-88% vs. LPS-activated conditions; Dunnett's multiple comparison test: p = 0.001).

[0108] The multiplex assay confirmed the ELISA results, i.e., TNF-α levels were reduced (-51.2%) in LPS-activated human macrophages in the presence of ATT (Fig. 1B). Furthermore, ATT significantly reduced IL-10 production (-60.3%) (Fig. 1C).

[0109] conclusion TNF-α is a pleiotropic molecule known to be involved in many pathological conditions, including vasculitis such as Kawasaki disease (Shen et al., 2013. Biochem Biophys Res Commun. 437(2):250-5).

[0110] The present results demonstrate that ATT can reduce cytokine release from activated human macrophages, clearly suggesting the possible therapeutic effect of ATT on vasculitis and Kawasaki disease in particular.

[0111] Example 2: Evaluation of ATT on vascular reactivity in the presence of endothelial dysfunction material and method The first order intrapulmonary artery (IPA1) of Wistar rats (male, 300 g) was divided into short tubular segments with an outer diameter of approximately 1.5-2 mm and used for isometric contraction measurements.

[0112] The arterial rings were placed in an isolated organ bath system (EMKA TECHNOLOGIES) containing Krebs solution (118 mM NaCl, 4.7 mM KCl, 1.2 mM MgSO, 2.5 mM NaHCO, 1.2 mM KHPO, 2.5 mM CaCl, and 11 mM D-glucose, pH 7.4) at 37°C and bubbled continuously with 15% O / 5% CO.

[0113] Arterial rings were stretched according to their diameter to a basal tension of 0.8-1.0 g. The tissue was equilibrated in Krebs solution for 1 h and washed out every 15 min. A high KCl solution (80 mM) was applied to obtain a reference contraction used to normalize subsequent contractile responses.

[0114] At the end of the experiment, endothelial function was tested for each ring by examining relaxation induced by carbamylcholine, a potent vasorelaxant (100 μM; Sigma), on precontracted pulmonary artery rings induced by 1 μM or 30 nM Phe. Passive and active mechanical properties were assessed using a transducer system connected to IOX2 software (v2.10.8, EMKA TECHNOLOGIES, Paris, France) to facilitate data acquisition and analysis. Data were then analyzed using two-way ANOVA with Bonferroni post-hoc tests (Prism 5 v.5.01).

[0115] Contractile function Contractile responses were tested by constructing cumulative concentration-response curves (CCRCs) to 5-hydroxytryptamine [5-HT] (10 nM–100 μM; Sigma). When indicated, tissues were preincubated with ATT (10 μM) for 30 min, and then CCRCs to 5-HT were constructed in the presence and absence of the drug.

[0116] N to inhibit nitric oxide-dependent relaxation ωAssessment of the role of the endothelium on the ATT effect was performed using ‐nitro‐L‐arginine methyl ester [L‐NAME] (100 μM, 30 min preincubation; Sigma) or 3‐[(3-cholamidopropyl)dimethylammonio]‐1‐propanesulfonate [CHAPS] (0.3% in water, perfused before mounting; Sigma) to physically damage the endothelium.

[0117] Relaxation of blood vessels Relaxation was assessed by injecting ATT (10 μM) into pulmonary artery rings precontracted with phenylephrine [Phe] (30 nM; Sigma) when contraction had reached its steady state.

[0118] The role of the endothelium on the ATT relaxation effect was evaluated as described previously by L-NAME preincubation or CHAPS perfusion. Endothelial function was tested for each ring by examining relaxation induced by 100 μM carbamylcholine (Sigma) on precontracted pulmonary artery rings induced by 1 μM or 30 nM Phe. Passive and active mechanical properties were evaluated using a transducer system coupled to IOX2 software (v2.10.8, EMKA technologies, Paris, France) to facilitate data acquisition and analysis. Data were then analyzed using two-way ANOVA with Bonferroni post-hoc tests (Prism 5v.5.01).

[0119] chronic endothelial dysfunction To examine the protective effect of ATT against chronic endothelial injury, rats were housed in a hypobaric hypoxic chamber (50 kPa or 10% O2) for 3 weeks.

[0120] During total hypoxia, continuous therapy was induced by a minipump placed intraperitoneally to deliver ATT (0.7 mg / mL) or its placebo (sulfobutylether-β-cyclodextrin).

[0121] After euthanasia of the animals, pulmonary artery rings were placed in the organ bath and vascular reactivity was examined by using cumulative concentration-response curves with 5-HT (10 nM to 100 μM). This ex vivo part of the study was performed without adding any ATT to the organ bath, meaning that the effects are only relevant to chronic in vivo treatment with the minipump.

[0122] result Protective effect of ATT on contractile function in the presence of endothelial dysfunction As shown in Figure 2A, the concentration-response curve in the presence of CHAPS-induced endothelial dysfunction was shifted to the left, indicating that the rings were more sensitive to 5-HT activation. Moreover, the maximum 5-HT availability was significantly enhanced by CHAPS incubation.

[0123] Surprisingly, the concentration-response curves and maximum efficacy in the presence of ATT and CHAPS were not significantly different from those obtained in the presence of normal endothelial function, thus demonstrating that ATT was able to prevent endothelial damage induced by CHAPS and restore normal contractility of the rings.

[0124] As shown in FIG. 2B, the concentration-response curve in the presence of L-NAME-induced endothelial dysfunction was also shifted to the left, meaning that the rings were more sensitive to 5-HT activation.

[0125] In the presence of ATT, the concentration-response curve with L-NAME was significantly shifted and the maximal effect was significantly reduced, demonstrating that ATT could prevent the alteration of the nitric oxide relaxation pathway induced by L-NAME and suppress the excessive contraction of the rings induced by 5-HT.

[0126] Protective effect of ATT on vascular relaxation in the presence of endothelial dysfunction The amplitude of relaxation to ATT alone or in the presence of L-NAME or CHAPS was expressed as a percentage of the maximal contraction to 30 nM phenylephrine normalized to 100% in control conditions or in the presence of L-NAME or CHAPS, respectively. As shown in Figure 3, the amplitude of relaxation was similar for groups of rings treated with ATT or CHAPS+ATT.

[0127] Surprisingly, pretreatment with L-NAME or CHAPS to induce endothelial dysfunction failed to affect the relaxation mediated by ATT (Figure 3, L-NAME+ATT and CHAPS+ATT, respectively). On the contrary, the relaxation induced by carbamylcholine under similar conditions was completely abolished by endothelial dysfunction induced by L-NAME or CHAPS (data not shown).

[0128] Taken together, these results demonstrate that ATT-induced vasorelaxation is not affected by endothelial dysfunction induced by CHAPS or L-NAME.

[0129] In vivo protective effect of ATT on contractile function in the presence of chronic endothelial dysfunction As shown in Figure 4, hypoxia-induced chronic endothelial dysfunction resulted in 5-HT-induced vascular hyperreactivity. Chronic treatment with ATT completely abolished this 5-HT-induced vascular hyperactivity, thereby confirming the protective effect of ATT against endothelial dysfunction in an in vivo model.

[0130] conclusion Vasculitic syndromes, including Kawasaki disease, are known to be associated with cardiovascular symptoms and complications, particularly arterial vascular insufficiency.

[0131] The present results demonstrate that ATT can suppress hyperconstriction and maintain vascular relaxation in arterial vessels with endothelial dysfunction, thus clearly suggesting the possible therapeutic effect of ATT on vasculitis and Kawasaki disease in particular.

[0132] Example 3: Evaluation of ATT on an in vivo mouse model of Kawasaki Disease Lactobacillus casei is a Gram-positive bacterium that colonizes the gastrointestinal and genitourinary tracts of both humans and animals. Cardiovascular lesions induced in mice by Lactobacillus casei cell wall extract (LCWE) are histologically similar to those observed in human vasculitis syndromes. LCWE-induced Kawasaki disease is characterized by infiltration of innate immune cells in the aortic root, development of necrotizing arteritis in the coronary arteries, and subsequent luminal obstruction that may lead to complete coronary stenosis.

[0133] Thus, the LCWE mouse model closely mimics the histopathological and immunopathological features of cardiovascular lesions in KD and has been used in evaluating the efficacy of drug candidates currently being tested in clinical trials (Lau et al., 2009. Clin Exp Immunol. 157(2):300-9; Gorelik et al., 2019. Clin Exp Immunol. 198(1):101-110).

[0134] Figure 5 shows the protocol performed on the LCWE mouse model to evaluate the effect of ATT on LCWE-induced KD.

[0135] Example 4: Evaluation of ATT on an in vitro human model of Kawasaki Disease material and method Adult human primary ventricular cardiomyocytes were isolated from two healthy donors by AnaBios (San Diego, USA) using their proprietary protocol, CardioPRIME™.

[0136] These isolated cardiomyocytes were exposed for 12 h to serum obtained from severely ill (2–6 days) Kawasaki disease patients with or without the addition of 1 μM ATT. In parallel, cardiomyocytes were exposed under the same conditions to serum from Wegener's disease patients or convalescent Kawasaki disease patients.

[0137] The CardioPRIME™ protocol implemented on the IonOptix MultiCell platform allows several parameters to be measured: upon electrical field stimulation to induce contraction (tracked by shape changes in individual cardiomyocytes under optical bright field microscopy), contractile transients could be analyzed for changes in contraction under different conditions (sarcomere shortening amplitude, peak height, maximum sarcomere departure velocity, maximum sarcomere return velocity and decay times to 30% and 90% relaxation).

[0138] result As shown in Figures 6A-6F, exposure to serum from Kawasaki disease patients for 12 hours reduced the systolic function and slowed the rate of relaxation of human cardiomyocytes.

[0139] ATT was found to prevent this loss of contractility (FIGS. 6A-6C), and also the loss of relaxation (FIGS. 6D-6F).

[0140] These parameters were consistent with the contraction and relaxation parameters of cardiomyocytes exposed to serum from convalescent Kawasaki disease patients, in which the patient's macrophages are no longer considered activated.

[0141] Conversely, when exposed to serum from Wegener's disease patients, cardiomyocytes did not show any specific changes during contraction and relaxation, suggesting different physiopathological characteristics of Kawasaki disease compared with Wegener's disease.

[0142] Overall, these data suggest therapeutic and prophylactic interest of ATT for vasculitis involving macrophage activation, hyperconstriction of arterial vessels with endothelial dysfunction (especially arterial vessels with reduced endothelial function), and / or vasoconstriction of arterial vessels, such as Kawasaki disease.

Claims

1. A pharmaceutical composition for use in preventing, treating, and / or reducing the severity or progression of Kawasaki disease or Kawasaki disease-like disorders in a subject in need thereof, the pharmaceutical composition comprising anethole trithione (ATT) or a structural analog thereof.

2. The pharmaceutical composition according to claim 1, wherein the preventing, treating, and / or reducing the severity or progression comprises reducing or inhibiting macrophage activation.

3. The pharmaceutical composition according to claim 1, wherein the preventing, treating, and / or reducing the severity or progression comprises reducing or inhibiting macrophage activation, and reducing or inhibiting macrophage activation comprises reducing the release of tumor necrosis factor (TNF)-α by macrophages.

4. The pharmaceutical composition according to claim 1, wherein the preventing, treating, and / or reducing the severity or progression comprises suppressing the contractility of arterial blood vessels having endothelial dysfunction, and / or preventing vasoconstriction of arterial blood vessels, and / or promoting vasodilation of arterial blood vessels.

5. The pharmaceutical composition according to claim 1, wherein the preventing, treating, and / or reducing the severity or progression comprises suppressing the contractility of arterial blood vessels with reduced endothelial function.

6. The pharmaceutical composition according to claim 1, wherein the disease is Kawasaki disease, Kawasaki disease-like disorder, or Kawasaki disease-like disorder associated with viral infection.

7. The pharmaceutical composition according to claim 1, wherein the disease is Kawasaki disease-like disorder associated with viral infection, and the viral infection is selected from the group consisting of SARS-CoV-2 infection, cytomegalovirus infection, Epstein-Barr virus infection, and human immunodeficiency virus infection.

8. The pharmaceutical composition according to claim 1, wherein the disease is Kawasaki disease-like disorder in a pediatric patient with COVID-19.

9. The pharmaceutical composition according to claim 1, wherein ATT is administered before, simultaneously with, or after a further therapeutic agent.

10. The pharmaceutical composition according to claim 1, wherein ATT is administered before, simultaneously with, or after a further therapeutic agent, and the further therapeutic agent is selected from the group consisting of intravenous immunoglobulin (IVIg), aspirin, corticosteroid, immunosuppressant, and alkylating agent.

11. The pharmaceutical composition according to claim 1, wherein the ATT is administered before, simultaneously with, or after a further therapeutic agent, and the further therapeutic agent is IVIg.

12. The pharmaceutical composition according to claim 1, wherein the subject is a pediatric or adolescent.

13. The structural analog of the ATT is of formula (I): 【Chemical 1】 (wherein, X represents S, O or NHOH, Y represents CH, C or N, R1, R ２ , R ４ and R ５ each independently represents hydrogen, hydroxy, halo, amino, alkylsulfonyl, aminosulfonyl, cyano, nitro, carboxy, aryl, alkoxy, haloalkyl, alkylamino, aminoalkyl, nitrooxyalkyl or carboxyalkyl, R ３ is methoxy or hydroxy, or R ３ and R ２ together with the carbon atom to which they are attached form a 5-membered heteroaryl moiety, where -R ３ -R ２ - is -A-CR ６ =B- or -B=CR ６ -A-, in the formula, A represents O, S or NR ７ wherein R ７ represents hydrogen, C １ to C ８ alkyl or alkyloxycarbonyl, B represents CH or N, and R ６ represents hydrogen, hydroxy, halo, amino, alkylsulfonyl, aminosulfonyl, cyano, nitro, carboxy, aryl, alkoxy, haloalkyl, alkylamino, aminoalkyl, nitrooxyalkyl or carboxyalkyl), is a compound, and the pharmaceutical composition according to any one of claims 1 to 12.

14. A pharmaceutical composition comprising anethole trithione (ATT) or a structural analog thereof for use in preventing, treating, and / or reducing the severity or progression of vasculitis in a subject in need thereof, wherein preventing, treating, and / or reducing the severity or progression of the disease comprises: (i) reducing or inhibiting macrophage activation, particularly reducing the release of tumor necrosis factor (TNF)-α by the macrophages, and / or (ii) suppressing the contractility of arterial vessels with endothelial dysfunction, and / or (iii) preventing vasoconstriction of arterial vessels, and / or (iv) promoting vasodilation of arterial vessels, and / or (v) suppressing the contractility of arterial vessels with reduced endothelial function.

15. The pharmaceutical composition according to claim 14, wherein the vasculitis is selected from the group consisting of Kawasaki disease, Kawasaki-like disease, giant cell arteritis, Takayasu arteritis, polyarteritis nodosa, antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis, microscopic polyangiitis, Wegener's disease, Churg-Strauss syndrome, anti-glomerular basement membrane (GBM) disease, Henoch-Schönlein purpura, cryoglobulinemic vasculitis, hypocomplementemic urticarial vasculitis, thrombotic occlusive vasculitis, Behçet's disease, Cogan's disease, cutaneous leukocytoclastic vasculitis, cutaneous arteritis, primary central nervous system vasculitis, localized arteritis, lupus vasculitis, rheumatoid vasculitis, sarcoid vasculitis, hepatitis C virus-related cryoglobulinemic vasculitis, hepatitis B virus-related vasculitis, syphilis-related aortitis, drug-related immune complex vasculitis, drug-related ANCA-associated vasculitis, and cancer-related vasculitis.

16. The pharmaceutical composition according to claim 14, wherein the vasculitis is granulomatous vasculitis. ​

17. The vasculitis is granulomatous vasculitis, and the granulomatous vasculitis is selected from the group consisting of Kawasaki disease, Takayasu arteritis, ANCA-associated vasculitis, Churg-Strauss syndrome, and primary central nervous system vasculitis. The pharmaceutical composition according to claim 14.

18. A pharmaceutical composition for use in preventing, treating, and / or reducing the severity or progression of a vasoconstrictive disorder in a subject in need thereof, the pharmaceutical composition comprising anethole trithione (ATT) or a structural analog thereof.

19. The vasoconstrictive disorder is selected from the group consisting of coronary artery vasospasm, vasospastic angina, diabetic neuropathy, diabetic gangrene, vascular headache, cerebral vasospasm, reversible cerebral vasospasm syndrome (RCVS), Raynaud's disease, acrocyanosis, Burger's disease, complex regional pain syndrome (CRPS), livedo reticularis, seizures, and post-traumatic dystrophy. The pharmaceutical composition according to claim 18.

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