CS585 (5-((4-((5,6-diphenylpyrazin-2-yl)(isopropyl)amino)butyl)thio)-2,4-dihydro-3h-1,2,4-triazol-3-one) for use in treating inflammatory and autoimmune disorders

Abstract

Disclosed are methods of treating or preventing an inflammatory or autoimmune disorder, such as antiphospholipid syndrome (''APS''), in a patient, comprising administering to the patient a therapeutically effective amount of Compound A, or a pharmaceutically acceptable salt thereof, or a prodrug thereof: (Compound A). Also disclosed are methods of increasing the level of cAMP in one or more neutrophils of a patient, comprising administering to the patient a therapeutically effective amount of Compound A, or a pharmaceutically acceptable salt thereof, or a prodrug thereof, and methods of reducing the level or preventing the formation of neutrophil extracellular traps (NETs) in a patient, comprising administering to the patient a therapeutically effective amount of Compound A, or a pharmaceutically acceptable salt thereof, or a prodrug thereof.

Description

30275 / 2025-132 UM 2025-132ANTIPLATELET COMPOUNDS FOR TREATING INFLAMMATORY AND AUTOIMMUNE DISORDERSBACKGROUNDTechnical Field

[0001] The present disclosure relates to inhibitors of platelet function, and methods of using the inhibitors to treat inflammatory and / or autoimmune disorders, such as antiphospholipid syndrome ("APS”).Description of Related Technology

[0002] Antiphospholipid syndrome (APS) is a leading acquired cause of thrombosis, impacting at least 1 in 2000 individuals. Mechanistically, APS is believed to be propelled by circulating antiphospholipid (aPL) antibodies that engage phospholipids and phospholipid-binding proteins at cell surfaces to activate the endothelium, neutrophils, and platelets— thereby tipping circulating blood toward thrombosis.

[0003] Vitamin K antagonists are the only treatment proven to reduce thrombosis in APS; however, despite their use, one in five patients will still experience a recurrent thrombotic event. Anticoagulants also do little to mitigate the small-vessel vasculopathy that leads to organ (e.g., brain, heart, kidney) deterioration over time.SUMMARY

[0004] There exists a need for therapeutic compounds capable of treating inflammatory and / or autoimmune disorders, and in particular inflammatory and / or autoimmune disorders characterized by thrombosis, such as APS, including anticoagulant-resistant manifestations of such inflammatory and / or autoimmune disorders.

[0005] Provided herein are methods of treating or preventing an inflammatory or autoimmune disorder in a patient, comprising administering to the patient a therapeutically effective amount of Compound A, or a pharmaceutically acceptable salt thereof, or a prodrug thereof:(Compound A). Also provided is a compound of formula A, a pharmaceutically acceptable salt thereof, or a prodrug thereof:use in the treatment or prevention of an inflammatory or autoimmune disorder in a patient. In various embodiments, treating or preventing the inflammatory or autoimmune disorder comprises increasing neutrophil cAMP formation, activating protein kinase A, reducing platelet activation, or any combination thereof, in the patient. In various embodiments, treating or preventing the inflammatory or autoimmune disorder comprises reducing neutrophil activation in the patient. In various30275 / 2025-132 UM 2025-132 embodiments, treating or preventing the inflammatory or autoimmune disorder comprises reducing the level or preventing the formation of neutrophil extracellular traps (NETs) in the patient., e.g., reducing or preventing NETosis in the patient.

[0006] Also provided are methods of increasing the level of cAMP in one or more neutrophils of a patient, comprising administering to the patient a therapeutically effective amount of Compound A, or a pharmaceutically acceptable salt thereof, or a prodrug thereof. Also provided is compound of formula A, a pharmaceutically acceptable salt thereof, or a prodrug thereof for use in increasing the level of cAMP in one or more neutrophils of a patient. In various embodiments, increasing the level of cAMP in one or more neutrophils of the patient treats or prevents an inflammatory or autoimmune disorder in the patient. In various embodiments, increasing the level of cAMP in one or more neutrophils of the patient comprises activating protein kinase A, reducing platelet activation, or both, in the patient. In various embodiments, increasing the level of cAMP in one or more neutrophils of the patient comprises reducing neutrophil activation in the patient. In various embodiments, increasing the level of cAMP in one or more neutrophils of the patient comprises reducing the level or preventing the formation of neutrophil extracellular traps (NETs) in the patient, e.g., reducing or preventing NETosis in the patient.

[0007] Also provided are methods of reducing the level or preventing the formation of neutrophil extracellular traps (NETs) in a patient, comprising administering to the patient a therapeutically effective amount of Compound A, or a pharmaceutically acceptable salt thereof, or a prodrug thereof. Also provided is compound of formula A, a pharmaceutically acceptable salt thereof, or a prodrug thereof for use in reducing the level or preventing the formation of neutrophil extracellular traps (NETs) in a patient. In various embodiments, reducing the level or preventing the formation of neutrophil extracellular traps (NETs) in the patient treats or prevents an inflammatory or autoimmune disorder in the patient. In various embodiments, reducing the level or preventing the formation of neutrophil extracellular traps (NETs) the patient comprises reducing or preventing NETosis in the patient.

[0008] In various embodiments, the inflammatory or autoimmune disorder is a clotting disorder. In various embodiments, the inflammatory or autoimmune disorder is characterized by thrombosis. In various embodiments, the inflammatory or autoimmune disorder is selected from the group consisting of antiphospholipid syndrome (APS), heparin-induced thrombocytopenia and thrombosis (HIT or HITT), vaccine-induced thrombocytopenia and thrombosis (VITT), immune thrombocytopenic purpura (ITP), claudication, and peripheral artery disease (PAD).BRIEF DESCRIPTION OF THE FIGURES

[0009] FIG. 1 depicts A) platelet cAMP determined in washed platelets of 20 healthy controls and 38 APS patients; B) activation of platelets with control or APS IgG in the presence of cAMP or cAMP-inducing agents; C) cAMP levels in neutrophils collected from 3-4 unique scleroderma patients pre- and post-infusion of the platelet inhibitor epoprostenol (prostacyclin); and D) LPS-induced NETosis in neutrophils collected from 3-4 unique scleroderma patients pre- and post-epoprostenol infusion. *p<0.05, **p<0.01, ***p<0.001, and ****p<0.0001 by unpaired or paired t-test.30275 / 2025-132 UM 2025-132

[0010] FIG. 2 shows that Compound A alters cytokines following daily oral dosing in mouse for 4 days. A) and B) show that Compound A reduces thrombotic mass and size, respectively.

[0011] FIG. 3 shows that Compound A alters cytokines following daily oral dosing in mouse for 4 days. A) shows that Compound A attenuates the inflammasome profile, and B) shows that Compound A attenuates cytokine profile.

[0012] FIG. 4 shows that Compound A alters cytokines following daily oral dosing in mouse for 4 days, A), B), and C) show that Compound significantly decreased important marker of inflammation (Ptgs2, Mefv, Irf3, respectively).

[0013] FIG. 5 shows the formation of neutrophil / platelet aggregates in the presence of thrombin.

[0014] FIG. 6 shows the formation of neutrophil / platelet aggregates with thrombin in the presence ofCompound A.

[0015] FIG. 7 shows a composite of the results from FIG. 6. Neutrophil / platelet active aggregates dose- dependently decreased with thrombin treatment in the presence of Compound A (upper right quadrant). N=4; ns=not significant, ***, P<0.001; ****, P<0.0001.

[0016] FIG. 8 shows the formation of neutrophil / platelet aggregates with control IgG and APS IgG.

[0017] FIG. 9 shows a composite of the results from FIG. 8. Neutrophil / platelet active aggregates dose- dependently decrease with APS IgG treatment in the presence of Compound A. N=4; ***, P<0.001; ****, P<0.0001.

[0018] FIG. 10 shows that neutrophil NETosis is prevented with Compound A treatment.DETAILED DESCRIPTION

[0019] Disclosed herein are treatments of inflammatory and / or autoimmune disorders characterized by thrombosis using a disclosed therapeutic agent.

[0020] Neutrophil extracellular traps (NETs)— tangles of DNA and microbicidal proteins expelled from activated neutrophils via a cell death program known as NETosis— are required for APS-associated thrombosis. At the same time, smoldering platelet activation in APS is associated with an increased risk of thrombosis. Inhibiting the excessive neutrophil and platelet activation inherent to inflammatory and / or autoimmune disorders characterized by thrombosis, e.g., APS, allows for more effective treatment of their anticoagulant-resistant features.

[0021] Boosting neutrophil cAMP with either adenosine receptor agonists or phosphodiesterase inhibitors restrains APS-associated NETosis, and these approaches are believed to be similarly effective in blunting platelet activation in inflammatory and / or autoimmune disorders characterized by thrombosis, such as APS.

[0022] 5-((4-((5,6-diphenylpyrazin-2-yl)(isopropyl)amino)butyl)thio)-2,4-dihydro-3H-1,2,4-triazol-3-one, referred to herein as "Compound A”, is an orally available, potent, selective, and stable prostacyclin receptor agonist drug candidate that inhibits platelet activation. Compound A is a synthesized analog of the oxylipin 12(S)-hydroxy-30275 / 2025-132 UM 2025-132 eicosatrienoic acid (12-HETrE), an endogenous regulator of platelet function that targets the prostacyclin receptor. Compound A has the following structure:Compound A

[0023] Without wishing to be bound by any particular theory, it is believed that both 12-HETrE and Compound A increase cAMP formation, activate protein kinase A, and attenuate platelet signaling, and that such effects will likely extend to neutrophils. Without wishing to be bound by any particular theory, it is believed that Compound A's antiplatelet and antineutrophil properties make it especially effective for attenuating the immunothrombosis inherent to inflammatory and / or autoimmune disorders characterized by thrombosis, like APS, without impacting coagulation or bleeding.

[0024] As pointed out above, this disclosure provides methods of treating or preventing inflammatory and / or autoimmune disorders characterized by thrombosis using a disclosed therapeutic agent.

[0025] More specifically, provided are methods of treating or preventing an inflammatory or autoimmune disorder in a patient, comprising administering to the patient a therapeutically effective amount of Compound A, or a pharmaceutically acceptable salt thereof, or a prodrug thereof:Compound A

[0026] As used herein, the term "inflammatory disorder” refers to a disorder characterized by an inflammatory response that does not return to a normal state. An inflammatory disorder may be an autoinflammatory disorder characterized by malfunction of the innate immune system. As used herein, the term "autoimmune disorder” refers to a (inflammatory) disorder characterized by malfunction of the adaptive immune system. In some cases, the disorder is an inflammatory disorder. In some cases, the disorder is an autoimmune disorder.

[0027] In some cases, the inflammatory or autoimmune disorder is a clotting disorder. As used herein, the term "clotting disorder” refers to a disorder characterized by predisposition to excessive thrombus formation. In some cases, the inflammatory or autoimmune disorder is characterized by thrombosis. In some cases, the30275 / 2025-132 UM 2025-132 inflammatory or autoimmune disorder is selected from the group consisting of antiphospholipid syndrome (APS), heparin-induced thrombocytopenia and thrombosis (HIT or HITT), vaccine-induced thrombocytopenia and thrombosis (VITT), immune thrombocytopenic purpura (ITP), claudication, or peripheral artery disease (PAD). In some cases, the inflammatory or autoimmune disorder is antiphospholipid syndrome (APS). In some cases, the inflammatory or autoimmune disorder is heparin-induced thrombocytopenia and thrombosis (HIT or HITT). In some cases, the inflammatory or autoimmune disorder is vaccine-induced thrombocytopenia and thrombosis (VITT). In some cases, the inflammatory or autoimmune disorder is immune thrombocytopenic purpura (ITP). In some cases, the inflammatory or autoimmune disorder is claudication. In some cases, the inflammatory or autoimmune disorder is peripheral artery disease (PAD).

[0028] In some cases, treating or preventing the inflammatory or autoimmune disorder comprises increasing neutrophil cAMP formation, activating protein kinase A, reducing platelet activation, or any combination thereof, in the patient. In some cases, treating or preventing the inflammatory or autoimmune disorder comprises increasing neutrophil cAMP formation in the patient. In some cases, treating or preventing the inflammatory or autoimmune disorder comprises activating protein kinase A in the patient. In some cases, treating or preventing the inflammatory or autoimmune disorder comprises reducing platelet activation in the patient. In some cases, treating or preventing the inflammatory or autoimmune disorder comprises increasing neutrophil cAMP formation and activating protein kinase A in the patient. In some cases, treating or preventing the inflammatory or autoimmune disorder comprises increasing neutrophil cAMP formation and reducing platelet activation in the patient. In some cases, treating or preventing the inflammatory or autoimmune disorder comprises activating protein kinase A and reducing platelet activation in the patient. In some cases, treating or preventing the inflammatory or autoimmune disorder comprises increasing neutrophil cAMP formation, activating protein kinase A, and reducing platelet activation in the patient.

[0029] In some cases, treating or preventing the inflammatory or autoimmune disorder comprises reducing neutrophil activation in the patient. In some cases, treating or preventing the inflammatory or autoimmune disorder comprises reducing the level or preventing the formation of neutrophil extracellular traps (NETs) in the patient. In some cases, treating or preventing the inflammatory or autoimmune disorder comprises reducing the level of neutrophil extracellular traps (NETs) in the patient. In some cases, treating or preventing the inflammatory or autoimmune disorder comprises preventing the formation of neutrophil extracellular traps (NETs) in the patient. In some cases, treating or preventing the inflammatory or autoimmune disorder comprises reducing or preventing NETosis in the patient.

[0030] Also provided are methods of increasing the level of cAMP in one or more neutrophils of a patient, comprising administering to the patient a therapeutically effective amount of Compound A, or a pharmaceutically acceptable salt thereof, or a prodrug thereof:30275 / 2025-132 UM 2025-132Compound A

[0031] In some cases, increasing the level of cAMP in one or more neutrophils of the patient treats or prevents an inflammatory or autoimmune disorder in the patient. In some cases, increasing the level of cAMP in one or more neutrophils of the patient treats or prevents an inflammatory disorder in the patient. In some cases, increasing the level of cAMP in one or more neutrophils of the patient treats or prevents an autoimmune disorder in the patient.

[0032] In some cases, the inflammatory or autoimmune disorder is a clotting disorder. In some cases, the inflammatory or autoimmune disorder is characterized by thrombosis. In some cases, the inflammatory or autoimmune disorder is selected from the group consisting of antiphospholipid syndrome (APS), heparin-induced thrombocytopenia and thrombosis (HIT or HITT), vaccine-induced thrombocytopenia and thrombosis (VITT), immune thrombocytopenic purpura (ITP), claudication, and peripheral artery disease (PAD). In some cases, the inflammatory or autoimmune disorder is antiphospholipid syndrome (APS). In some cases, the inflammatory or autoimmune disorder is heparin-induced thrombocytopenia and thrombosis (HIT or HITT). In some cases, the inflammatory or autoimmune disorder is vaccine-induced thrombocytopenia and thrombosis (VITT). In some cases, the inflammatory or autoimmune disorder is immune thrombocytopenic purpura (ITP). In some cases, the inflammatory or autoimmune disorder is claudication. In some cases, the inflammatory or autoimmune disorder is peripheral artery disease (PAD).

[0033] In some cases, increasing the level of cAMP in one or more neutrophils of the patient comprises activating protein kinase A, reducing platelet activation, or both, in the patient. In some cases, increasing the level of cAMP in one or more neutrophils of the patient comprises activating protein kinase A in the patient. In some cases, increasing the level of cAMP in one or more neutrophils of the patient comprises reducing platelet activation in the patient. In some cases, increasing the level of cAMP in one or more neutrophils of the patient comprises activating protein kinase A and reducing platelet activation in the patient.

[0034] In some cases, increasing the level of cAMP in one or more neutrophils of the patient comprises reducing neutrophil activation in the patient. In some cases, increasing the level of cAMP in one or more neutrophils of the patient comprises reducing the level or preventing the formation of neutrophil extracellular traps (NETs) in the patient. In some cases, increasing the level of cAMP in one or more neutrophils of the patient comprises reducing the level of neutrophil extracellular traps (NETs) in the patient. In some cases, increasing the level of cAMP in one or more neutrophils of the patient comprises preventing the formation of neutrophil30275 / 2025-132 UM 2025-132 extracellular traps (NETs) in the patient. In some cases, increasing the level of cAMP in one or more neutrophils of a patient comprises reducing or preventing NETosis in the patient.

[0035] Also provided are methods of reducing the level or preventing the formation of neutrophil extracellular traps (NETs) in a patient, comprising administering to the patient a therapeutically effective amount of Compound A, or a pharmaceutically acceptable salt thereof, or a prodrug thereof:Compound A

[0036] In some cases, reducing the level or preventing the formation of neutrophil extracellular traps (NETs) in the patient treats or prevents an inflammatory or autoimmune disorder in the patient. In some cases, reducing the level or preventing the formation of neutrophil extracellular traps (NETs) in the patient treats or prevents an inflammatory disorder in the patient. In some cases, reducing the level or preventing the formation of neutrophil extracellular traps (NETs) in the patient treats or prevents an autoimmune disorder in the patient. In some cases, reducing the level of neutrophil extracellular traps (NETs) in the patient treats or prevents an inflammatory or autoimmune disorder in the patient. In some cases, reducing the level of neutrophil extracellular traps (NETs) in the patient treats or prevents an inflammatory disorder in the patient. In some cases, reducing the level of neutrophil extracellular traps (NETs) in the patient treats or prevents an autoimmune disorder in the patient. In some cases, preventing the formation of neutrophil extracellular traps (NETs) in the patient treats or prevents an inflammatory or autoimmune disorder in the patient. In some cases, preventing the formation of neutrophil extracellular traps (NETs) in the patient treats or prevents an inflammatory disorder in the patient. In some cases, preventing the formation of neutrophil extracellular traps (NETs) in the patient treats or prevents an autoimmune disorder in the patient.

[0037] In some cases, the inflammatory or autoimmune disorder is a clotting disorder. In some cases, the inflammatory or autoimmune disorder is characterized by thrombosis. In some cases, the inflammatory or autoimmune disorder is selected from the group consisting of antiphospholipid syndrome (APS), heparin-induced thrombocytopenia and thrombosis (HIT or HITT), vaccine-induced thrombocytopenia and thrombosis (VITT), immune thrombocytopenic purpura (ITP), claudication, and peripheral artery disease (PAD). In some cases, the inflammatory or autoimmune disorder is antiphospholipid syndrome (APS). In some cases, the inflammatory or autoimmune disorder is heparin-induced thrombocytopenia and thrombosis (HIT or HITT). In some cases, the inflammatory or autoimmune disorder is vaccine-induced thrombocytopenia and thrombosis (VITT). In some cases, the inflammatory or autoimmune disorder is immune thrombocytopenic purpura (ITP). In some cases, the30275 / 2025-132 UM 2025-132 inflammatory or autoimmune disorder is claudication. In some cases, the inflammatory or autoimmune disorder is peripheral artery disease (PAD).

[0038] In some cases, reducing the level or preventing the formation of neutrophil extracellular traps (NETs) in the patient comprises reducing or preventing NETosis in the patient. In some cases, reducing the level of neutrophil extracellular traps (NETs) in the patient comprises reducing or preventing NETosis in the patient. In some cases, preventing the formation of neutrophil extracellular traps (NETs) in the patient comprises reducing or preventing NETosis in the patient.

[0039] In any of the methods disclosed herein, Compound A, the salt, or the prodrug thereof can be administered in a dosage of 0.1 to 500 mg / kg. In some cases, Compound A, the salt, or the prodrug thereof is administered in a dosage of 0.1-500 mg / kg. In some cases, Compound A, the salt, or the prodrug thereof is administered orally.Compound A

[0040] As used herein the term "Compound A” refers to 5-((4-((5,6-diphenylpyrazin-2- yl)(isopropyl)amino)butyl)thio)-2,4-dihydro-3H-1,2,4-triazol-3-one, having the following structure:Compound A

[0041] The synthesis and chemical characterization of Compound A has been described in the art, e.g., in WO 2019 / 204447 A1 . Compound A is also known as "CS585”.

[0042] Compound A or its pharmaceutically acceptable salts and prodrugs described herein (termed the "therapeutic agent”) can be administered to a patient in a therapeutically effective amount (e.g., in an amount sufficient to prevent or relieve the symptoms of an inflammatory or autoimmune disorder, such as APS). The therapeutic agent can be administered alone or as part of a pharmaceutically acceptable composition. The therapeutic agent can be administered all at once, multiple times, or delivered substantially uniformly over a period of time.Salts

[0043] To practice the disclosed methods, a pharmaceutically acceptable salt of Compound A can be used. As used herein, the term "pharmaceutically acceptable salt” refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, which is incorporated30275 / 2025-132 UM 2025-132 herein by reference. Pharmaceutically acceptable salts of the compound above include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable, nontoxic acid addition salts are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, trifluoroacetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, glutamate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy- ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2- naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3- phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p- toluenesulfonate, undecanoate, valerate salts, and the like. Salts of compounds containing a carboxylic acid or other acidic functional group can be prepared by reacting with a suitable base. Such salts include, but are not limited to, alkali metal, alkaline earth metal, aluminum salts, ammonium, N+(Ci-4alky 1)4 salts, and salts of organic bases such as trimethylamine, triethylamine, morpholine, pyridine, piperidine, picoline, dicyclohexylamine, N,N'- dibenzylethylenediamine, 2-hydroxyethylamine, bis-(2-hydroxyethyl)amine, tri-(2-hydroxyethyl)amine, procaine, dibenzylpiperidine, dehydroabietylamine, N, N'-bisdehydroabietylamine, glucamine, N-methylglucamine, collidine, quinine, quinoline, and basic amino acids such as lysine and arginine. This disclosure also envisions the quaternization of any basic nitrogen-containing groups of the compound provided herein. Water or oil-soluble or dispersible products may be obtained by such quaternization. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, lower alkyl sulfonate and aryl sulfonate. Exemplary pharmaceutically acceptable salts of Compound A include, but are not limited to, hydrochloride, hydrobromide, sulfate, nitrate, phosphate, mesylate, esylate, isethionate, tosylate, napsylate, besylate, acetate, propionate, maleate, benzoate, salicylate, fumerate, glutamate, aspartate, citrate, lactate, succinate, tartrate, glycollate, hexanoate, octanoate, decanoate, oleate, stearate, pamoate, and polystryrene sulfonate.Prodruqs

[0044] In some embodiments, a prodrug of Compound A can also be used. As used herein, the term "prodrug” refers to a medication or compound that, after administration, is metabolized (i.e., converted within the body) into a pharmacologically active drug. Thus, a prodrug of Compound A refers to a medication or compound that, after administration, is converted within the body into Compound A. To a skilled artisan, a corresponding prodrug can be used instead to improve how a medicine is absorbed, distributed, metabolized, and excreted, thereby improving the pharmacological effect exerted in a subject (e.g., a human) in need thereof. Examples of a prodrug that can be used in this invention include, but are not limited to, amides, carbamates, sulfonamides,30275 / 2025-132 UM 2025-132 carboxamides, N-oxides, N-acyloxyalkyl derivatives, N-hydroxyalkyl derivatives, and N-(phosphoryloxy)alkyl derivatives.Dosing and Formulation

[0045] A skilled artisan will appreciate that the dosage of the therapeutic agent can be varied over time. A particular administration regimen for a particular subject will depend, in part, upon the compound, the amount of compound administered, the route of administration, and the cause and extent of any side effects. The amount of compound administered to a patient (e.g., a mammal, such as a human) in accordance with the disclosure should be sufficient to effect the desired response over a reasonable time frame. Dosage typically depends upon the route, timing, and frequency of administration. Accordingly, the clinician titers the dosage and modifies the route of administration to obtain the optimal therapeutic effect, and conventional range-finding techniques are known to those of ordinary skill in the art.

[0046] Purely by way of illustration, the above-described method of treating inflammatory and / or autoimmune disorders comprises administering, e.g., from about 0.1 mg / kg up to about 500 mg / kg of Compound A or a doseequivalent amount of a salt or prodrug of Compound A (i.e., an amount of a salt or prodrug of Compound A having the same therapeutic effect as the recited dose of Compound A), depending on the factors mentioned above. In other embodiments, the dosage ranges can be from 5 mg / kg up to about 100 mg / kg; or 10 mg / kg up to about 100 mg / kg, or 10 mg / kg up to about 60 mg / kg, or 20 mg / kg up to about 40 mg / kg. Some conditions require prolonged treatment, which may or may not entail administering lower doses of compound over multiple administrations. If desired, a dosage of the compound is administered as two, three, four, five, six or more subdoses administered separately at appropriate intervals throughout the day, optionally, in unit dosage forms. The treatment period will depend on the particular condition, and may last one day to several months.

[0047] Suitable methods of administering a pharmaceutically acceptable composition comprising Compound A are well known in the art. Although more than one route can be used to administer a compound, a particular route can provide a more immediate and more effective reaction than another route. Depending on the circumstances, a pharmaceutical composition comprising the compound is applied or instilled into body cavities, absorbed through the skin or mucous membranes, ingested, inhaled, and / or introduced into circulation. For example, in certain circumstances, it will be desirable to deliver a pharmaceutical composition comprising the therapeutic agent orally, through injection, or by one of the following means: intravenous, intraperitoneal, intracerebral (intra-parenchymal), intracerebroventricular, intramuscular, intra-ocular, intraarterial, intraportal, intralesional, intramedullary, intrathecal, intraventricular, transdermal, subcutaneous, intraperitoneal, intranasal, enteral, topical, sublingual, urethral, vaginal, or rectal. The compound can be administered by sustained release systems, or by implantation devices.

[0048] To facilitate administration, the therapeutic agent is, in various aspects, formulated into a pharmaceutically acceptable composition comprising a carrier (e.g., vehicle, adjuvant, or diluent). The particular carrier employed is limited only by chemico-physical considerations, such as solubility and lack of reactivity with the compound, and by the route of administration. Pharmaceutically acceptable carriers are well known in the art.30275 / 2025-132 UM 2025-132Illustrative pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions (for example, see U.S. Patent No. 5,466,468). Injectable compositions are further described in, e.g., Pharmaceutics and Pharmacy Practice, J. B. Lippincott Co., Philadelphia. Pa., Banker and Chalmers, eds., pages 238-250 (1982), and ASHP Handbook on Injectable Drugs, Toissel, 4th ed., pages 622-630 (1986)). A pharmaceutical composition comprising the therapeutic agent is, in one aspect, placed within containers, along with packaging material that provides instructions regarding the use of such pharmaceutical compositions. Generally, such instructions include a tangible expression describing the reagent concentration, as well as, in certain embodiments, relative amounts of excipient ingredients or diluents (e.g., water, saline or PBS) that may be necessary to reconstitute the pharmaceutical composition.

[0049] Compositions suitable for parenteral injection may comprise physiologically acceptable sterile aqueous or nonaqueous solutions, dispersions, suspensions, or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Examples of suitable aqueous and nonaqueous carriers, diluents, solvents, or vehicles include water, ethanol, polyols (propylene glycol, polyethylene glycol, glycerol, and the like), suitable mixtures thereof, vegetable oils (such as olive oil) and injectable organic esters such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.

[0050] These compositions may also contain adjuvants such as preserving, wetting, emulsifying, and dispersing agents. Microorganism contamination can be prevented by adding various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, and the like. It may also be desirable to include isotonic agents, for example, sugars, sodium chloride, and the like. Prolonged absorption of injectable pharmaceutical compositions can be brought about by the use of agents delaying absorption, for example, aluminum monostearate and gelatin.

[0051] Solid dosage forms for oral administration include capsules, tablets, powders, and granules. In such solid dosage forms, the therapeutic agent is admixed with at least one inert customary excipient (or carrier) such as sodium citrate or dicalcium phosphate or (a) fillers or extenders, as for example, starches, lactose, sucrose, mannitol, and silicic acid; (b) binders, as for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and acacia; (c) humectants, as for example, glycerol; (d) disintegrating agents, as for example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solution retarders, as for example, paraffin; (f) absorption accelerators, as for example, quaternary ammonium compounds; (g) wetting agents, as for example, cetyl alcohol and glycerol monostearate; (h) adsorbents, as for example, kaolin and bentonite; and (i) lubricants, as for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, and tablets, the dosage forms may also comprise buffering agents. Solid compositions of a similar type may also be used as fillers in soft and hard filled gelatin capsules using such excipients as lactose or milk sugar, as well as high molecular weight polyethylene glycols, and the like.30275 / 2025-132 UM 2025-132

[0052] Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells, such as enteric coatings and others well known in the art. The solid dosage forms may also contain opacifying agents. Further, the solid dosage forms may be embedding compositions, such that they release the therapeutic agent in a certain part of the intestinal tract in a delayed manner. Examples of embedding compositions that can be used are polymeric substances and waxes. Compound A can also be in micro- encapsulated form, optionally with one or more excipients.

[0053] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs. In addition to the therapeutic agent, the liquid dosage form may contain inert diluents commonly used in the art, such as water or other solvents, solubilizing agents and emulsifiers, as for example, ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 ,3-butylene glycol, dimethylformamide, oils, in particular, cottonseed oil, groundnut oil, corn germ oil, olive oil, castor oil, and sesame seed oil, glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, or mixtures of these substances, and the like.

[0054] Besides such inert diluents, the composition can also include adjuvants, such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents. Suspensions, in addition to the therapeutic agent, may contain suspending agents, as for example, ethoxylated isosteary I alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar, and tragacanth, or mixtures of these substances, and the like.

[0055] Upon formulation, solutions can be administered in a manner compatible with the dosage formulation and in such amount as is therapeutically effective. The compositions are easily administered in a variety of dosage forms such as injectable solutions, drug release capsules and the like. For parenteral administration in an aqueous solution, for example, the solution should be suitably buffered if necessary and the liquid diluent first rendered isotonic with sufficient saline or glucose. These particular aqueous solutions are especially suitable for intravenous, intramuscular, subcutaneous and intraperitoneal administration.Methods of Treatment and Uses

[0056] Compound A can treat inflammatory and / or autoimmune disorders in a unique way. Compound A has been demonstrated to inhibit platelet activation by activating the G protein-coupled receptor known as the prostacyclin (IP) receptor. Activation of the IP receptor results in formation of cAMP in the cell through activation of adenylyl cyclase which converts ATP to cAMP.

[0057] cAMP in the platelet results in inhibition of platelet activation and thrombus formation in the vessel, which is a promising therapeutic strategy for treating inflammatory and / or autoimmune disorders characterized by thrombosis, such as APS. APS results from a decrease in cAMP in immune cells and is correlated to platelet activation and thrombosis of the vasculature. Increasing the levels of cAMP in immune cells prevents their activation and formation of neutrophil extracellular nets (NETosis) which in turn increases the likelihood of platelet activation and thrombosis. APS affects at least 1 in 2000 individuals in the US. Beyond clinical events, APS criteria seek the stable presence of anticardiolipin or anti-beta-2 glycoprotein I Q32GPI) antibodies; without30275 / 2025-132 UM 2025-132 wishing to be bound by any particular theory, the latter are likely particularly pathogenic. Further, the "lupus anticoagulant” test, which screens for aPL antibodies based on prolongation of phospholipid-dependent clotting times, detects a variety of aPL antibodies, including anti-p2GPI and anti-phosphatidylserine / prothrombin (PS / PT) antibodies.

[0058] Anticoagulation is the only treatment known to reduce the vascular complications of APS. While such regimens are somewhat effective in preventing large-vessel events, such as deep vein thrombosis (DVT) and stroke, one in five patients will still experience a breakthrough thrombotic event. Anticoagulants also do little to mitigate the chronic small-vessel disease that leads to organ (e.g., brain, heart, kidneys) deterioration.

[0059] p2GPI associates with apolipoprotein E receptor 2 (ApoER2) and glycoprotein lb (GPIb) on the platelet surface, creating platforms for anti-p2GPI antibodies to trigger platelet activation. Anti-PS / PT antibodies also activate platelets. Unlike for neutrophils, Fey receptors appear to be required for aPL-mediated platelet activation. Mitogen-activated protein (MAP) kinases and mammalian target of rapamycin (mTOR) pathway signaling also play a role. Without wishing to be bound by any particular theory, it is believed that NET- associated DNA triggers coagulation, while NET-associated histones activate endothelial cells and platelets. Past work has revealed that patient-derived anti-p2GPI antibodies activate NETosis via Toll-like receptor and beta-integrin signaling, and that thrombosis can be restrained in APS mouse models by (I) boosting intracellular cAMP to block NETosis or (II) combatting NETs that have already been released with DNases or polyanions.

[0060] The only current treatment are anticoagulants, and this approach is only effective in 80% of patients and only for limiting the thrombotic disease in large vessels. Small vessel thrombosis, however, is not decreased with anticoagulants and is the cause of multi-organ dysfunction over time. Data in the Examples of the present disclosure demonstrate that Compound A administered to mice once per day for 4 days by oral administration (oral gavage) results in inhibition of immune cell activity and repression of the cytokine inflammatory pathways. Hence, treatment with Compound A represents the first viable treatment for this rare disease.

[0061] The use of Compound A to treat other inflammatory and autoimmune diseases, such as those characterized by thrombus formation, is also provided herein. In some cases, the inflammatory or autoimmune disorder is a clotting disorder. In some cases, the inflammatory or autoimmune disorder is characterized by thrombosis. In some cases, the inflammatory or autoimmune disorder is selected from the group consisting of antiphospholipid syndrome (APS), heparin-induced thrombocytopenia and thrombosis (HIT or HITT), vaccine- induced thrombocytopenia and thrombosis (VITT), immune thrombocytopenic purpura (ITP), claudication, and peripheral artery disease (PAD). In some cases, the inflammatory or autoimmune disorder is antiphospholipid syndrome (APS). In some cases, the inflammatory or autoimmune disorder is heparin-induced thrombocytopenia and thrombosis (HIT or HITT). In some cases, the inflammatory or autoimmune disorder is vaccine-induced thrombocytopenia and thrombosis (VITT). In some cases, the inflammatory or autoimmune disorder is immune thrombocytopenic purpura (ITP). In some cases, the inflammatory or autoimmune disorder is claudication. In some cases, the inflammatory or autoimmune disorder is peripheral artery disease (PAD).30275 / 2025-132 UM 2025-132

[0062] As used herein, the term "treat,” as well as words related thereto, do not necessarily imply 100% or complete treatment. Rather, there are varying degrees of treatment of which one of ordinary skill in the art recognizes as having a potential benefit or therapeutic effect. In this respect, the methods of treating inflammatory and / or autoimmune disorders of the present disclosure can provide any amount or any level of treatment of the inflammatory and / or autoimmune disorders. Furthermore, the treatment provided by the methods of the present disclosure may include treatment of one or more conditions or symptoms of the inflammatory and / or autoimmune disorders, being treated. Also, the treatment provided by the methods of the present disclosure may encompass slowing the progression of the inflammatory and / or autoimmune disorders. For example, the disclosed methods can treat inflammatory and / or autoimmune disorders by virtue of preventing thrombus formation, reducing thrombus formation, reducing thrombus size, prevention of NETosis, and the like.

[0063] Also provided herein are prophylactic methods, i.e., methods of preventing inflammatory and / or autoimmune disorders of the present disclosure in a patient. As used herein, the term "preventing” refers to administration of a composition disclosed herein (e.g., Compound A, a pharmaceutically-acceptable salt thereof, or a prodrug thereof) which reduces the frequency of, or delays the onset of, symptoms of a medical condition in a patient relative to a patient which does not receive the composition. Thus, prevention of an inflammatory or autoimmune disorder characterized by thrombosis includes, for example, reducing the number of thrombi in a population of patients receiving a prophylactic treatment relative to an untreated control population, and / or delaying the appearance of detectable thrombi in a treated population versus an untreated control population, e.g., by a statistically and / or clinically significant amount. Thus, also contemplated is the use of a composition disclosed herein (e.g., Compound A, a pharmaceutically-acceptable salt thereof, or a prodrug thereof) for increasing the level of cAMP in one or more neutrophils of a patient and / or reducing the level or preventing the formation of neutrophil extracellular traps (NETs) in a patient in a prophylactic manner (e.g., to reduce and / or delay the appearance of thrombi in the patient). Such prophylactic use can therefore prevent or attenuate the occurrence of inflammatory and / or autoimmune disorders of the present disclosure in the patient.

[0064] In some cases, prophylactic methods disclosed herein are used to prevent inflammatory and / or autoimmune disorders of the present disclosure in a patient belonging to an at-risk population, i.e., a population having a predisposition to having or developing one or more inflammatory and / or autoimmune disorders of the present disclosure. For example, APS is three to five times more common in female patients than male patients, more common in patients having a family history of APS, and is more common in patients having another inflammatory and / or autoimmune disorder (such as lupus). Accordingly, such patients are said to at-risk populations for having or developing APS.

[0065] Also contemplated are prophylactic methods for maintenance in inflammatory and / or autoimmune disorders, e.g., for preventing recurrence or maintaining remission of such a disorder in a patient that has previously been treated for the disorder and has since entered remission from the disorder.

[0066] In some cases, the prophylactic methods disclosed herein are useful for preventing the development of an inflammatory and / or autoimmune disorder (e.g., a clotting disorder) that is secondary to a primary condition in a patient. For example, the methods are useful for preventing the development of APS as a secondary disorder30275 / 2025-132 UM 2025-132 in a patient experiencing pregnancy or lupus as a primary condition. Non-limiting examples of primary conditions which may lead to secondary inflammatory and / or autoimmune disorders that may be prevented by the methods disclosed herein include cancer, diabetes, vaccines, and infection.

[0067] In jurisdictions that forbid the patenting of methods that are practiced on the human body, the meaning of "administering” of a composition to a human subject or patient shall be restricted to prescribing a controlled substance that a human subject or patient will self-administer by any technique (e.g., orally, inhalation, topical application, injection, insertion, etc.). The broadest reasonable interpretation that is consistent with laws or regulations defining patentable subject matter is intended. In jurisdictions that do not forbid the patenting of methods that are practiced on the human body, the "administering” of compositions includes both methods practiced on the human body and also the foregoing activities.

[0068] The embodiments also relate to Compound A, a pharmaceutically acceptable salt thereof, or a prodrug thereof, for use in the treatment or preventions of an inflammatory or autoimmune disorder in a patient as disclosed herein. The embodiments further relate to use of Compound A, a pharmaceutically acceptable salt thereof, or a prodrug thereof, in the manufacture of a medicament for treating or preventing an inflammatory or autoimmune disorder in a patient. Also provided are Compound A, a pharmaceutically acceptably salt thereof, or a prodrug thereof, for use in increasing the level of cAMP in one or more neutrophils of a patient, and the use of Compound A, a pharmaceutically acceptable salt thereof, or a prodrug thereof, in the manufacture of a medicament for increasing the level of cAMP in one or more neutrophils of a patient. Also provided are Compound A, a pharmaceutically acceptable salt thereof, or a prodrug thereof, for use in reducing the level or preventing the formation of NETs in a patient and the use of Compound A, a pharmaceutically acceptable salt thereof, or a prodrug thereof, in the manufacture of a medicament for reducing the level or preventing the formation of NETs in a patient.Kits

[0069] Also provided herein are kits that include a pharmaceutical formulation comprising a compound described herein, such as Compound A and instructions for administering the pharmaceutical formulation to a patient. In some embodiments the kit is provided with a device for administering the formulation to a patient. The kit may further include a variety of containers, e.g., vials, tubes, bottles, and the like. In various embodiments, the kit comprises a syringe and a needle, wherein the formulation is optionally prepackaged within the syringe. In various embodiments, the kit comprises a prepackaged or predosed oral formulation of Compound A and optionally a device for administering the oral formulation.

[0070] Without further elaboration, it is believed that one skilled in the art can, based on the above description, utilize the present disclosure to its fullest extent. The following specific examples, i.e., EXAMPLES 1-4, are therefore to be construed as merely illustrative, and not limitative of the remainder of the disclosure in any way whatsoever.30275 / 2025-132 UM 2025-132EXAMPLESExample 1 : Synthesis and characterization of 5-((4-((5,6-diphenylpyrazin-2-yl)(isopropyl)amino)butyl)thio)-2,4- dihydro-3H-1 ,2,4-triazol-3-one (Compound A)5 Compound A

[0071] Reagents and conditions: (a) Platinum(IV) oxide, EtOH, ambient temp, 7 days; (b) 5-bromo-2,3- diphenylpyrazine, KI, 140°C, 2 days; (c) DMAP, 4-methylbenzenesulfonyl chloride, DCM, ambient temp, 18 hrs;(d) 5-mercapto-2,4-dihydro-3H-1,2,4-triazol-3-one, NaH, DMF, ambient temp, 2 days.

[0072] 4-(isopropylamino)butan-1-ol (Intermediate 3). To a solution of 4-aminobutan-1-ol (2.7 ml, 29 mmol, 2) in acetone (3.5 ml, 47mmol, 1) was added platinum(IV) oxide (67 mg, 0.29 mmol). The mixture stirred under H2 atmosphere at ambient temperature for 7 days. The reaction mixture was filtered through Celite, and concentrated under reduced pressure to afford Intermediate 3.

[0073] 4-((5,6-diphenylpyrazin-2-yl)(isopropyl)amino)butan-1-ol (Intermediate 4). To a mixture of 5-bromo-2,3- diphenylpyrazine (1 g, 3.21 mmol) and 4-(isopropylamino)butan-1-ol (2.32 g, 17.7 mmol, Intermediate 3) was added potassium iodide (266 mg, 1.6 mmol). The mixture was stirred in a pressure vessel at 140°C for 2 days. The reaction mixture was cooled to room temp and diluted with water. The mixture was extracted with EtOAc (x4) and the combined organic layers washed with brine, dried over Na2SO4, filtered and concentrated. The crude residue was purified with column chromatography eluting with 0-2% MeOH in DCM. The fractions of the desired product were combined, concentrated, and dried under reduced pressure to obtain Intermediate 4.

[0074] N-(4-chlorobutyl)-N-isopropyl-5, 6-diphenylpyrazin-2-amine (Intermediate 5). To a solution of 4-((5,6- diphenylpyrazin-2-yl)(isopropyl)amino)butan-1 -ol (200 mg, 0.55 mmol, Intermediate 4) and dry DCM (8 ml) was added 4-methylbenzenesulfonyl chloride (320 mg, 1.68 mmol) and DMAP (210 mg, 1.72 mmol). The reaction was stirred at ambient temperature for 18 hrs. The reaction mixture was diluted with water and extracted with DCM (x3). The organic layer was washed with brine, dried over Na2SO4, filtered and concentrated. The crude residue was purified with column chromatography eluting with DCM. The fractions of desired product were30275 / 2025-132 UM 2025-132 combined, concentrated, and dried under reduced pressure to yield Intermediate 5. MS (ESI), m / z (%): 380 (M+, 100%).

[0075] 5-((4-((5, 6-diphenylpyrazin-2-yl) (isopropyl)amino)butyl) thio)-2, 4-dihydro-3H-1 ,2, 4-triazol-3-one (Compound A). To a solution of 5-mercapto-2,4-dihydro-3H-1,2,4-triazol-3-one (92.5 mg, 0.79 mmol) and DMF (2 ml) in 0°C ice bath was added NaH (55 mg, 1.38 mmol). N-(4-chlorobutyl)-N-isopropyl-5,6-diphenylpyrazin-2- amine (150 mg, 0.4 mmol, Intermediate 5) was dissolved in DMF (9 ml) and added to the mixture. The reaction was stirred at ambient temperature for 2 days. The reaction mixture was quenched with ice water and extracted with EtOAc (x3). The pH of the aqueous layer was adjusted to 7 with 2N HCI, followed by extraction with EtOAc (x2). The organic layer was washed with brine, dried over Na2SO4, filtered and concentrated. The crude residue was purified with column chromatography eluting 2% MeOH in DCM. The fractions of the desired product were combined, concentrated and dried under reduced pressure to yield Compound A. MS (ESI), m / z (%): 461 (M+, 100%).Example 2 - Increasing cAMP levels in APS with prostacyclin receptor agonists

[0076] Intracellular cAMP and platelet activation: FIG 1A illustrates the cAMP levels determined in platelets from healthy subjects (control) and in platelets from APS patients (APS). The cAMP is assessed in platelets from control and APS patients using a cAMP ELISA kit whereby cAMP can quantitively be assessed using a specific OD on a spectrophotometer. These numbers are aligned to a standard cAMP curve acquired from the same plasma samples. As is shown in FIG 1A, APS platelets have low baseline cAMP levels as compared to control platelets from healthy subjects.

[0077] Increasing cAMP levels prevents APS IgG-mediated platelet activation: In FIG 1B, P-selectin (marker of platelet activation) was measured by flow cytometry using the P-selectin antibody CD62P. As a reference, a control IgG was added to plasma to attain a basal or reference P-selectin level (control IgG). APS IgG was added to plasma to attain the APS-dependent level of P-selectin (alpha granule secretion) levels on the surface of the platelet (APS IgG). APS IgG platelet rich plasma was treated with an inducer of cAMP production in the form of an adenosine receptor agonist (A2AR), direct addition of cAMP (dibut-cAMP or 8-Br-cAMP), or direct activation of adenylate cyclase (AC, an enzyme catalyzing the generation of cAMP from ATP) with forskolin all result in a decrease in P-selectin expression on the surface of the platelets and thereby a decrease in platelet activation.

[0078] NETosis and prostacyclin receptor agonists: Scleroderma patients receive infusions of prostacyclin receptor agonists for refractory finger ulcerations. As compared with neutrophils prior to epoprostenol infusion (pre) (n=3-4 unique patients), neutrophils following epoprostenol infusion (post) have more cAMP as measured in vitro in a cAMP assay (FIG 1C) and a blunted ability to undergo NETosis as measured in a NETosis assay (FIG 1D). Since NETosis is the result of neutrophil activation and externalization of its DNA to the blood, it plays a central role in clot formation and thrombosis. Inhibiting neutrophil activation through increasing the cAMP in the cell prevents NETosis, which in turn reduces the likelihood and severity of thrombosis in the vessel.Example 3 - Immune cell cytokine profile of Compound A vs. control30275 / 2025-132 UM 2025-132

[0079] Compound A altered cytokine profiles following daily oral dosing in mouse for 4 days. Mice were treated with Compound A once per day for 4 days following ligation to measure the ability of Compound A to attenuate venous thrombosis. The size and mass of the clots were assessed after four days of ligation and dosing. The clot size and mass were reduced in mice treated with Compound A (FIG 2A, 2B). Additionally, the inflammasome profile (FIG 3A) and innate and adaptive immunity profile (FIG 3B) of monocytes and neutrophils was assessed using a multiplexing and gene analysis approach from these mice following the procedure. The inflammasome and cytokine profiles were attenuated in the immune cells isolated from animals treated with Compound A compared to animals treated with vehicle (15% DMSO, 35% PEG, and 50% saline) (FIG 3A, 3B). A number of important markers of inflammation including Ptgs2, Mefv, and Irf3, were observed to be significantly decreased in immune cells from animals treated daily with Compound A (FIG 4A-4C).Example 4 - Evaluation of intracellular cAMP and platelet activation

[0080] A longitudinal cohort comprising a number (e.g., approximately 250) of patients with durably positive aPL antibodies according to developed ACR / EULAR criteria are sampled. Biospecimens (plasma, serum, DNA, whole-blood RNA, neutrophil RNA) are collected at every clinic visit (every 3 to 6 months) and at the time of new thrombotic events.

[0081] Addition of patient plasma containing aPL antibodies is added to plasma from healthy subjects prior to and following addition of Compound A. The ability of Compound A to inhibit platelet activation in the presence of patient plasma is assessed in platelet rich plasma.

[0082] Procoagulant activity of the platelets is assessed by measuring surface exposure of phosphatidyl serine (PS) and P-selectin by flow cytometry measurement. This measurement is conducted in platelet rich plasma of healthy subjects that is pre-treated with Compound A followed by addition of aPL antibody-containing plasma from patients.

[0083] Platelet adhesion under arterial shear pressure is assessed in whole blood in a flow chamber coated with collagen. Under arterial flow 1,8001seconds, whole blood that is treated with either Compound A or vehicle followed by addition of patient plasma containing aPL antibodies is added to the assay along with calcein-AM to visualize the platelets. The blood flows over the collagen strip at arterial shear rates and platelet adhesion and clot formation is visualized and monitored in real-time with a high-speed fluorescent microscope.

[0084] Platelet adhesion is assessed using the Total Thrombus Activation System (T-TAS-01). Platelet adhesion under arterial shear pressure is assessed in whole blood in a T-TAS-01. Blood is collected in special T- TAS blood tubes and treated with or without Compound A. The blood is then treated with or without aPL antibody-containing plasma from patients and added to the T-TAS-01 system where pressure changes are measured as the T-TAS channel coated with collagen forms a clot. The time to clot and area under the curve are assessed to determine if the clotting is normal enhanced, or attenuated.

[0085] Platelet activation is assessed by measuring the activation of the al Ibb3 integrins as well as granule secretion by flow cytometry methods in whole blood. Platelets are treated with or without Compound A followed by treatment with or without aPL antibody-containing patient plasma and the level of integrin activation and30275 / 2025-132 UM 2025-132 granule secretion are measured with fluorescently-labeled antibodies. The integrin antibody is PAC1 which only binds to activated integrin allbb3. The alpha granule secretion is measured with CD62P antibody which recognizes P-selectin on the surface of the platelet. The Dense granule is measured with CD63 antibody which recognizes a protein specifically released from the dense granule.

[0086] FIGs. 5-9 show the results of neutrophil / platelet aggregation studies with thrombin. Both neutrophils (Y-axis in FIGs. 5, 6, and 8) and platelets (X-axis in FIGs. 5, 6, and 8) are activated by thrombin. FIG. 5 shows that neutrophil / platelet active aggregates increase with thrombin treatment (upper right quadrant), while FIG. 6 shows that neutrophil / platelet active aggregates dose-dependently decreased with thrombin treatment in the presence of Compound A (upper right quadrant of each chart). FIG. 7 shows a composite of the data in FIG. 6, with neutrophil / platelet aggregate (NPA) percentage shown on the Y-axis, and treatment method shown on the X-axis.

[0087] FIG. 8 shows the formation of neutrophil / platelet aggregates with control IgG and APS IgG. Neutrophil / platelet aggregates were assessed by flow cytometry (neutrophils: Y-axis; platelets: X-axis). Neutrophils were unstimulated (untreated) or activated by either control human IgG or IgG from patients with antiphospholipid syndrome (APS IgG) at a concentration of 50 pig / ml (top row). Neutrophil / platelet active aggregates were not observed to increase with control IgG. However, addition of APS IgG resulted in a significant increase in active neutrophil / platelet aggregates. Neutrophil / platelet aggregates pre-treated with Compound A (bottom row) showed no increase in neutrophil / platelet aggregates following addition of either control IgG or APS IgG (upper right quadrant of each chart). FIG. 9 shows a composite of the data in FIG. 8, with neutrophil / platelet aggregate (NPA) percentage shown on the Y-axis, and treatment method shown on the X-axis.

[0088] Platelet aggregation is assessed by measuring the degree of platelet-platelet interaction and clumping in vitro in a Chrono-LOG lumi-aggregometer. Platelet rich plasma from healthy subjects is placed in a stirring cuvette in the lumi-aggregometer. The platelets are treated with or without Compound A. aPL antibody is added to the cuvette and the degree of platelet aggregation is measured in real-time for up to 10 minutes.Identifying mechanisms by which prostacyclin receptor agonists combat APS-relevant neutrophil and platelet activation in vitro.

[0089] Baseline cAMP levels are determined in APS patient neutrophils and platelets (both low at baseline), and whether Compound A can boost them. The role of Compound A in preventing neutrophil and platelet activation in response to APS-relevant stimulation (anti-|32GPI, anti-PS / PT, etc.) is studied in at least 20 APS patients from the longitudinal cohort.In vivo models for Compound A’s APS-relevant properties extend to in vivo models.

[0090] Administration of Compound A to mice reduces APS patient IgG-accelerated venous and arterial thrombosis. For this model, APS patient-IgG is administered via intravenous administration to mice followed by oral administration of Compound A and induction of a venous thrombosis injury (electrolytic or ligation). APS patient-IgG accelerated venous thrombosis (where the role of NETosis is well established) is attenuated by oral30275 / 2025-132 UM 2025-132 administration of Compound A. Compound A's protective properties also extend to APS IgG-accelerated arterial thrombosis (where platelets are instrumental).

[0091] A cohort of 6-8 mice per group are used, which allows the detection of differences in mean venous thrombus weight of ~1 .5 mg (a=0.05, power=80%). Male and female mice are used in equal numbers, and post- hoc analyses assess for differences. Normally distributed data is analyzed by 2-tailed t-test or 1-way ANOVA corrected for multiple comparisons; skewed data are analyzed by Mann-Whitney or Kruskal-Wallis tests.Example 5 - Neutrophil NETosis is prevented with Compound A treatment

[0092] Human neutrophils were placed in cell culture dishes followed by addition of control IgG or APS IgG (FIG. 10, top row center and right, respectively). Neutrophils were exposed to DNA stain (bright points) to assess neutrophil extracellular traps (NETosis) formation. It was observed that 50 pig / ml of control IgG did not induce NETosis (FIG. 10, top center). However, addition of 50 pig / ml of APS IgG resulted in significant NETosis (FIG. 10, top right). The bottom row in FIG. 10 assessed if increasing concentrations of Compound A inhibited APS IgG-induced NETosis in neutrophils. While 10 nM Compound A did not inhibit APS IgG-induced NETosis, both 100 nM and 1 piM Compound A prevented APS IgG-induced NETosis in the human neutrophils.

[0093] Overall, these studies demonstrate that the activation of the IP receptor with Compound A results in cAMP formation in both neutrophils and platelets. Without wishing to be bound by any particular theory, cAMP is believed to prevent both neutrophil and platelet activation, aggregate formation, and NETosis. Thus, treatment with Compound A shows promise as an effective method for prevention or treatment of APS.OTHER EMBODIMENTS

[0094] All of the features disclosed in this specification may be combined in any combination. Each feature disclosed in this specification may be replaced by an alternative feature serving the same, equivalent, or similar purpose. Thus, unless expressly stated otherwise, each feature disclosed is only an example of a generic series of equivalent or similar features.

[0095] Further, from the above description, one skilled in the art can easily ascertain the essential characteristics of the present disclosure, and without departing from the spirit and scope thereof, can make various changes and modifications of the disclosure to adapt it to various usages and conditions. Thus, other embodiments are also within the claims.30275 / 2025-132 UM 2025-132REFERENCES1 Barbhaiy a, M. et al. 2023 ACR / EULAR Antiphospholipid Syndrome Classification Criteria. Arthritis Rheumatol, doi:10.1002 / art.42624 (2023).2 Duarte-Garcia, A. etal. The Epidemiology of Antiphospholipid Syndrome: A Population-Based Study.Arthritis Rheumatol 71, 1545-1552, doi:10.1002 / art.40901 (2019).3 Knight, J. S., Branch, D. W. & Ortel, T. L. Antiphospholipid syndrome: advances in diagnosis, pathogenesis, and management. BMJ 380, e069717, doi:10.1136 / bmj-2021-069717 (2023).4 Jackson, W. G. et al. Recurrent thrombosis in patients with antiphospholipid antibodies and arterial thrombosis on antithrombotic therapy. Blood Adv l, 2320-2324, doi : 10.1182 / bloodadvances.2017008185 (2017).5 Long, B. R. & Leya, F. The role of antiphospholipid syndrome in cardiovascular disease. Hematol Oncol Clin North Am 22, 79-94, vi-vii, doi:10.1016 / j.hoc.2007.10.002 (2008).6 Alarcon-Segovia, D., Cardiel, M. H. & Reyes, E. Antiphospholipid arterial vasculopathy. J Rheumatol 16, 762-767 (1989).7 Hughson, M. D., McCarty, G. A. & Brumback, R. A. Spectrum of vascular pathology affecting patients with the antiphospholipid syndrome. Hum Pathol 26, 716-724, doi:10.1016 / 0046-8177(95)90218-x (1995).8 Yalavarthi, S. etal. Release of neutrophil extracellular traps by neutrophils stimulated with antiphospholipid antibodies: a newly identified mechanism of thrombosis in the antiphospholipid syndrome. Arthritis Rheumatol 67, 2990-3003, doi: 10.1002 / art.39247 (2015).9 Meng, H. etal. In Vivo Role of Neutrophil Extracellular Traps in Antiphospholipid Antibody-Mediated Venous Thrombosis. Arthritis Rheumatol 69, 655-667, doi : 10.1002 / art.39938 (2017).10 All, R. A. et al. Adenosine receptor agonism protects against NETosis and thrombosis in antiphospholipid syndrome. Nat Commun 10, 1916, doi:10.1038 / s41467-019-09801-x (2019).11 All, R. A. et al. Defibrotide Inhibits Antiphospholipid Antibody-Mediated Neutrophil Extracellular Trap Formation and Venous Thrombosis. Arthritis Rheumatol 74, 902-907, doi : 10.1002 / art.42017 (2022).12 All, R. A. et al. Antineutrophil properties of natural gingerols in models of lupus. JCI Insight 6, doi : 10.1172 / jci .insight.138385 (2021 ).13 All, R. A. et al. Ginger intake suppresses neutrophil extracellular trap formation in autoimmune mice and healthy humans. JCI Insight 8, doi:10.1172 / jci.insight.172O11 (2023).14 Stanger, L. et al. The oxylipin analog CS585 prevents platelet activation and thrombosis through activation of the prostacyclin receptor. Blood 142, 1556-1569, doi: 10.1182 / blood.2023020622 (2023).15 Tourdot, B. E. et al. 12-HETrE inhibits platelet reactivity and thrombosis in part through the prostacyclin receptor. Blood Adv , 1124-1131, doi:10.1182 / bloodadvances.2017006155 (2017).16 Pengo, V. et al. Antiphospholipid syndrome: antibodies to Domain 1 of beta2-glycoprotein 1 correctly classify patients at risk. J Thromb Haemost 13, 782-787, doi:10.1111 / jth.12865 (2015).17 Andreoli, L. et al. Clinical characterization of antiphospholipid syndrome by detection of IgG antibodies against beta2 -glycoprotein I domain 1 and domain 4 / 5: ratio of anti-domain 1 to anti-domain 4 / 5 as a useful new biomarker for antiphospholipid syndrome. Arthritis Rheumatol 67, 2196-2204, doi:10.1002 / art.39187 (2015).18 McDonnell, T. C. R. et al. PEGylated Domain I of Beta-2-Glycoprotein I Inhibits the Binding, Coagulopathic, and Thrombogenic Properties of IgG From Patients With the Antiphospholipid Syndrome. Front ImmunoI Q, 2413, doi: 10.3389 / fimmu.2018.02413 (2018).19 Shi, H. et al. Antiphosphatidylserine / prothrombin antibodies (aPS / PT) as potential diagnostic markers and risk predictors of venous thrombosis and obstetric complications in antiphospholipid syndrome. Clin Chem Lab Med 56, 614-624, doi:10.1515 / cclm-2017-0502 (2018).20 von Bruhl, M. L. et al. Monocytes, neutrophils, and platelets cooperate to initiate and propagate venous thrombosis in mice in vivo. J Exp Med 209, 819-835, doi:10.1084 / jem.20112322 (2012).21 Gould, T. J. et al. Neutrophil extracellular traps promote thrombin generation through platelet-dependent and platelet-independent mechanisms. Arterioscler Thromb Vase Biol 34, 1977-1984, doi:10.1161 / ATVBAHA114.304114 (2014).22 Shi, H. et al. Endothelium-protective, histone-neutralizing properties of the polyanionic agent defibrotide. JCI Insight s, doi:10.1172 / jci.insight.149149 (2021).23 Fuchs, T. A. et al. Extracellular DNA traps promote thrombosis. Proceedings of the National Academy of Sciences of the United States of America 107, 15880-15885, doi: 10.1073 / pnas.1005743107 (2010).30275 / 2025-132 UM 2025-13224 Sule, G. et al. Increased Adhesive Potential of Antiphospholipid Syndrome Neutrophils Mediated by beta2 Integrin Mac-1. Arthritis Rheumatol 72, 114-124, doi: 10.1002 / art.41057 (2020).25 Tambralli, A., Gockman, K. & Knight, J. S. NETs in APS: Current Knowledge and Future Perspectives. Curr Rheumatol Rep 22, 67, doi: 10.1007 / s11926-020-00936-1 (2020).26 Knight, J. S. et al. Activated signature of antiphospholipid syndrome neutrophils reveals potential therapeutic target. JCI Insight2, doi: 10.1172 / jci.insight.93897 (2017).27 Pennings, M. T. et al. Platelet adhesion to dimeric beta-glycoprotein I under conditions of flow is mediated by at least two receptors: glycoprotein Ibalpha and apolipoprotein E receptor 2'. J Thromb Haemost 5, 369-377, doi: 10.1111 / j.1538-7836.2007.02310.x (2007).28 Chayoua, W. et al. Antiprothrombin antibodies induce platelet activation: A possible explanation for anti- FXa therapy failure in patients with antiphospholipid syndrome? J Thromb Haemost , 1776-1782, doi:10.1111 / jth.15320 (2021).29 Hol lerbach, A. et al. Platelet Activation by Antiphospholipid Antibodies Depends on Epitope Specificity and is Prevented by mTOR Inhibitors. Thromb Haemost 119, 1147-1153, doi:10.1055 / S-0039- 1685453 (2019).30 Vega-Ostertag, M., Harris, E. N. & Pierangeli, S. S. Intracellular events in platelet activation induced by antiphospholipid antibodies in the presence of low doses of thrombin. Arthritis Rheum 50, 2911-2919, doi:10.1002 / art.20434 (2004).

Claims

30275 / 2025-132 UM 2025-132What is Claimed:1 . A method of treating or preventing an inflammatory or autoimmune disorder in a patient, comprising administering to the patient a therapeutically effective amount of Compound A, or a pharmaceutically acceptable salt thereof, or a prodrug thereof:(Compound A).

2. The method of claim 1 , wherein the inflammatory or autoimmune disorder is a clotting disorder.

3. The method of claim 1 or 2, wherein the inflammatory or autoimmune disorder is characterized by thrombosis.

4. The method of any one of claims 1 to 3, wherein the inflammatory or autoimmune disorder is selected from the group consisting of antiphospholipid syndrome (APS), heparin-induced thrombocytopenia and thrombosis (HIT or HITT), vaccine-induced thrombocytopenia and thrombosis (VITT), immune thrombocytopenic purpura (ITP), claudication, and peripheral artery disease (PAD).

5. The method of claim 4, wherein the inflammatory or autoimmune disorder is antiphospholipid syndrome (APS).

6. The method of any one of claims 1 to 5, wherein treating or preventing the inflammatory or autoimmune disorder comprises increasing neutrophil cAMP formation, activating protein kinase A, reducing platelet activation, or any combination thereof, in the patient.

7. The method of any one of claims 1 to 6, wherein treating or preventing the inflammatory or autoimmune disorder comprises reducing neutrophil activation in the patient.

8. The method of any one of claims 1 to 7, wherein treating or preventing the inflammatory or autoimmune disorder comprises reducing the level or preventing the formation of neutrophil extracellular traps (NETs) in the patient.

9. The method of any one of claims 1 to 8, wherein treating or preventing the inflammatory or autoimmune disorder comprises reducing or preventing NETosis in the patient.

10. The method of any one of claims 1 to 9, wherein the Compound A, the salt, or the prodrug thereof is administered in a dosage of 0.1 to 500 mg / kg.11 . The method of claim 10, wherein the Compound A, the salt, or the prodrug thereof is administered in a dosage of 0.1 to 20 mg / kg.

12. The method of any one of claims 1 to 11 , wherein the Compound A, the salt, or the prodrug thereof is administered orally.30275 / 2025-132 UM 2025-13213. A method of increasing the level of cAMP in one or more neutrophils of a patient, comprising administering to the patient a therapeutically effective amount of Compound A, or a pharmaceutically acceptable salt thereof, or a prodrug thereof:(Compound A).

14. The method of claim 13, wherein increasing the level of cAMP in one or more neutrophils of the patient treats or prevents an inflammatory or autoimmune disorder in the patient.

15. The method of claim 14, wherein the inflammatory or autoimmune disorder is a clotting disorder.

16. The method of claim 14 or 15, wherein the inflammatory or autoimmune disorder is characterized by thrombosis.

17. The method of any one of claims 14 to 16, wherein the inflammatory or autoimmune disorder is selected from the group consisting of antiphospholipid syndrome (APS), heparin-induced thrombocytopenia and thrombosis (HIT or HITT), vaccine-induced thrombocytopenia and thrombosis (VITT), immune thrombocytopenic purpura (ITP), claudication, and peripheral artery disease (PAD).

18. The method of claim 17, wherein the inflammatory or autoimmune disorder is antiphospholipid syndrome (APS).

19. The method of any one of claims 13 to 18, wherein increasing the level of cAMP in one or more neutrophils of the patient comprises activating protein kinase A, reducing platelet activation, or both, in the patient.

20. The method of any one of claims 13 to 19, wherein increasing the level of cAMP in one or more neutrophils of the patient comprises reducing neutrophil activation in the patient.

21. The method of any one of claims 13 to 20, wherein increasing the level of cAMP in one or more neutrophils of the patient comprises reducing the level or preventing the formation of neutrophil extracellular traps (NETs) in the patient.

22. The method of any one of claims 13 to 21, wherein increasing the level of cAMP in one or more neutrophils of a patient comprises reducing or preventing NETosis in the patient.

23. The method of any one of claims 13 to 22, wherein the Compound A, the salt, or the prodrug thereof is administered in a dosage of 0.1 to 500 mg / kg.

24. The method of claim 23, wherein the Compound A, the salt, or the prodrug thereof is administered in a dosage of 0.1-500 mg / kg.30275 / 2025-132 UM 2025-13225. The method of any one of claims 13 to 24, wherein the Compound A, the salt, or the prodrug thereof is administered orally.

26. A method of reducing the level or preventing the formation of neutrophil extracellular traps (NETs) in a patient, comprising administering to the patient a therapeutically effective amount of Compound A, or a pharmaceutically acceptable salt thereof, or a prodrug thereof:(Compound A).

27. The method of claim 26, wherein reducing the level or preventing the formation of neutrophil extracellular traps (NETs) in the patient treats or prevents an inflammatory or autoimmune disorder in the patient.

28. The method of claim 27, wherein the inflammatory or autoimmune disorder is a clotting disorder.

29. The method of claim 27 or 28, wherein the inflammatory or autoimmune disorder is characterized by thrombosis.

30. The method of any one of claims 27 to 29, wherein the inflammatory or autoimmune disorder is selected from the group consisting of antiphospholipid syndrome (APS), heparin-induced thrombocytopenia and thrombosis (HIT or HITT), vaccine-induced thrombocytopenia and thrombosis (VITT), immune thrombocytopenic purpura (ITP), claudication, and peripheral artery disease (PAD).31 . The method of claim 30, wherein the inflammatory or autoimmune disorder is antiphospholipid syndrome (APS).

32. The method of any one of claims 26 to 31 , wherein reducing the level or preventing the formation of neutrophil extracellular traps (NETs) the patient comprises reducing or preventing NETosis in the patient.

33. The method of any one of claims 26 to 32, wherein the Compound A, the salt, or the prodrug thereof is administered in a dosage of 0.1 to 500 mg / kg.

34. The method of claim 33, wherein the Compound A, the salt, or the prodrug thereof is administered in a dosage of 0.1-500 mg / kg.

35. The method of any one of claims 26 to 34, wherein the Compound A, the salt, or the prodrug thereof is administered orally.

36. A compound of formula A, a pharmaceutically acceptable salt thereof, or a prodrug thereof:30275 / 2025-132 UM 2025-132A for use in the treatment or prevention of an inflammatory or autoimmune disorder in a patient.

37. The compound, salt, or prodrug for use of claim 36, wherein the inflammatory or autoimmune disorder is a clotting disorder.

38. The compound, salt, or prodrug for use of claim 36 or 37, wherein the inflammatory or autoimmune disorder is characterized by thrombosis.

39. The compound, salt, or prodrug for use of any one of claims 36 to 38, wherein the inflammatory or autoimmune disorder is selected from the group consisting of antiphospholipid syndrome (APS), heparin-induced thrombocytopenia and thrombosis (HIT or HITT), vaccine-induced thrombocytopenia and thrombosis (VITT), immune thrombocytopenic purpura (ITP), claudication, and peripheral artery disease (PAD).

40. The compound, salt, or prodrug for use of claim 39, wherein the inflammatory or autoimmune disorder is antiphospholipid syndrome (APS).41 . The compound, salt, or prodrug for use of any one of claims 36 to 40, wherein the treatment or prevention of the inflammatory or autoimmune disorder comprises increasing neutrophil cAMP formation, activating protein kinase A, reducing platelet activation, or any combination thereof, in the patient.

42. The compound, salt, or prodrug for use of any one of claims 36 to 41 , wherein the treatment or prevention of the inflammatory or autoimmune disorder comprises reducing neutrophil activation in the patient.

43. The compound, salt, or prodrug for use of any one of claims 36 to 42, wherein the treatment or prevention of the inflammatory or autoimmune disorder comprises reducing the level or preventing the formation of neutrophil extracellular traps (NETs) in the patient.

44. The compound, salt, or prodrug for use of any one of claims 36 to 43, wherein the treatment or prevention of the inflammatory or autoimmune disorder comprises reducing or preventing NETosis in the patient.

45. The compound, salt, or prodrug for use of any one of claims 36 to 44, formulated for administration in a dosage of 0.1 to 500 mg / kg.

46. The compound, salt, or prodrug for use of claim 45, formulated for administration in a dosage of 0.1-20 mg / kg.

47. The compound, salt, or prodrug for use of any one of claims 36 to 46, formulated for oral administration.30275 / 2025-132 UM 2025-13248. A compound of formula A, a pharmaceutically acceptable salt thereof, or a prodrug thereof:A for use in increasing the level of cAMP in one or more neutrophils of a patient.

49. The compound, salt, or prodrug for use of claim 48, wherein increasing the level of cAMP in one or more neutrophils of the patient treats or prevents an inflammatory or autoimmune disorder in the patient.

50. The compound, salt, or prodrug for use of claim 49, wherein the inflammatory or autoimmune disorder is a clotting disorder.51 . The compound, salt, or prodrug for use of claim 49 or 50, wherein the inflammatory or autoimmune disorder is characterized by thrombosis.

52. The compound, salt, or prodrug for use of any one of claims 49 to 51 , wherein the inflammatory or autoimmune disorder is selected from the group consisting of antiphospholipid syndrome (APS), heparin-induced thrombocytopenia and thrombosis (HIT or HITT), vaccine-induced thrombocytopenia and thrombosis (VITT), immune thrombocytopenic purpura (ITP), claudication, and peripheral artery disease (PAD).

53. The compound, salt, or prodrug for use of claim 52, wherein the inflammatory or autoimmune disorder is antiphospholipid syndrome (APS).

54. The compound, salt, or prodrug for use of any one of claims 48 to 53, wherein increasing the level of cAMP in one or more neutrophils of the patient comprises activating protein kinase A, activating platelet activation, or both, in the patient.

55. The compound, salt, or prodrug for use of any one of claims 48 to 54, wherein increasing the level of cAMP in one or more neutrophils of the patient comprises reducing neutrophil activation in the patient.

56. The compound, salt, or prodrug for use of any one of claims 48 to 55, wherein increasing the level of cAMP in one or more neutrophils of the patient comprises reducing the level or preventing the formation of neutrophil extracellular traps (NETs) in the patient.

57. The compound, salt, or prodrug for use of any one of claims 48 to 56, wherein increasing the level of cAMP in one or more neutrophils of a patient comprises reducing or preventing NETosis in the patient.

58. The compound, salt, or prodrug for use of any one of claims 48 to 57, formulated for administration in a dosage of 0.1 to 500 mg / kg.30275 / 2025-132 UM 2025-13259. The compound, salt, or prodrug for use of claim 58, formulated for administration in a dosage of 0.1-500 mg / kg.

60. The compound, salt, or prodrug for use of any one of claims 48 to 59, formulated for oral administration.61 . A compound of formula A, a pharmaceutically acceptable salt thereof, or a prodrug thereof:A for use in reducing the level or preventing the formation of neutrophil extracellular traps (NETs) in a patient.

62. The compound, salt, or prodrug for use of claim 61, wherein reducing the level or preventing the formation of neutrophil extracellular traps (NETs) in the patient treats or prevents an inflammatory or autoimmune disorder in the patient.

63. The compound, salt, or prodrug for use of claim 62, wherein the inflammatory or autoimmune disorder is a clotting disorder.

64. The compound, salt, or prodrug for use of claim 62 or 63, wherein the inflammatory or autoimmune disorder is characterized by thrombosis.

65. The compound, salt, or prodrug for use of any one of claims 62 to 64, wherein the inflammatory or autoimmune disorder is selected from the group consisting of antiphospholipid syndrome (APS), heparin-induced thrombocytopenia and thrombosis (HIT or HITT), vaccine-induced thrombocytopenia and thrombosis (VITT), immune thrombocytopenic purpura (ITP), claudication, and peripheral artery disease (PAD).

66. The compound, salt, or prodrug for use of claim 65, wherein the inflammatory or autoimmune disorder is antiphospholipid syndrome (APS).

67. The compound, salt, or prodrug for use of any one of claims 61 to 66, wherein reducing the level or preventing the formation of neutrophil extracellular traps (NETs) the patient comprises reducing or preventing NETosis in the patient.

68. The compound, salt, or prodrug for use of any one of claims 61 to 67, formulated for administration in a dosage of 0.1 to 500 mg / kg.

69. The compound, salt, or prodrug for use of claim 68, formulated for administration in a dosage of 0.1-500 mg / kg.30275 / 2025-132 UM 2025-13270. The compound, salt, or prodrug for use of any one of claims 61 to 69, formulated for oral administration.

Images