Dosing regimen of NLRP3 inhibitors

Abstract

The present disclosure relates to the field of pharmacy, in particular to NLRP3 inhibitors for use in the treatment of autoinflammatory syndrome. The disclosure also relates to an NLRP3 inhibitor or a pharmaceutical combination comprising an NLRP3 inhibitor or a pharmaceutically acceptable salt thereof and at least one additional therapeutic agent for use in the treatment of autoinflammatory syndrome; to a method for treating autoinflammatory syndrome, the method involving administration of an NLRP3 inhibitor or such a combination; and to the use of an NLRP3 inhibitor or such a combination for the manufacture of a medicament for the treatment of autoinflammatory syndrome. In particular, N '-((1, 2, 3, 5, 6, 7-hexahydro-symmetric indacen-4-yl) carbamoyl)-2-(2-hydroxypropyl-2-yl) thiazol-5-sulfonylimide amide and its enantiomers are used for the treatment of autoinflammatory syndrome, such as inflammatory syndrome, inflammatory syndrome, inflammatory syndrome, inflammatory syndrome, and the like. The present invention relates to compounds of formula (I), in particular, cryptic fever protein related periodic syndrome (CAPS), familial cold-type autoinflammatory syndrome (FCAS), Mueller-Wills syndrome (MWS), newborn diseased multi-system inflammatory disease / chronic infant neuroskin and joint syndrome (NOMIM / CINCA) or familial geothermal fever (FMF).

Description

Technical Field

[0001] The present disclosure relates to the field of pharmacy, and in particular to NLRP3 inhibitors for use in treating autoinflammatory syndromes. The present disclosure also relates to NLRP3 inhibitors for use in treating autoinflammatory syndromes or a pharmaceutical combination comprising an NLRP3 inhibitor or a pharmaceutically acceptable salt thereof and at least one additional therapeutic agent; to a method for treating autoinflammatory syndromes, the method involving administering an NLRP3 inhibitor or the combination; and to the use of an NLRP3 inhibitor or the combination for the manufacture of a medicament for treating autoinflammatory syndromes. Background Art

[0002] Cryptopyrin-associated periodic syndromes (CAPS) are a group of rare diseases characterized by cutaneous, musculoskeletal, ocular, and neurological symptoms and chronic systemic inflammation that can lead to organ damage and / or amyloidosis and are caused by heterozygous gain-of-function mutations in the nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3 (NLRP3) gene. CAPS is divided into three clinical phenotypes based on the spectrum of disease severity: familial cold autoinflammatory syndrome (FCAS), Muckler-Wills syndrome (MWS), and neonatal-onset multisystem inflammatory disease (NOMID), also known as chronic infantile neurocutaneous and articular syndrome (CINCA) (Hoffman et al., (2019) Cryopyrin-Associated Periodic Syndromes (CAPS). [Cryopyrin-associated periodic syndrome (CAPS)] This document is published in: Hashkes P, Laxer R, Simon A (eds.). Textbook of Autoinflammation. [Textbook of Autoinflammation] Springer, Cham. pp. 347-365).

[0003] FCAS represents the mildest clinical phenotype of CAPS, and symptoms are typically limited to low-grade fever, widespread rash, conjunctivitis, and polyarthralgias, which are triggered within 1-2 hours of cold exposure and resolve within 24 hours after warming. These clinical manifestations, episodes, or onsets typically begin in infancy and continue throughout life. Many patients with FCAS also show evidence of chronic inflammation between episodes, particularly a rash that occurs each afternoon, which can be associated with headaches, myalgias, and fatigue that occur in the evening, but chronic inflammation rarely leads to amyloidosis in this patient population (approximately 2%).

[0004] Familial Mediterranean fever (FMF) is caused by mutations in the MEFV gene and is the most common of all known autoinflammatory diseases. Mutations in the MEFV gene reduce the activity of the pyrin protein, similar to the NLRP3 gene in MWS, FCAS, and NOMID / CINCA, thereby disrupting the control of the inflammatory process. FMF is characterized by repeated attacks of painful inflammation in the abdomen, chest, or joints. These attacks are often accompanied by fever and sometimes rash or headache. Inflammation may occasionally occur in other parts of the body, such as the heart; the membranes around the brain and spinal cord; and in men, in the testicles. Typically, attacks last 12 to 72 hours and can vary in severity. The length of time between onset is also variable and can vary from a few days to several years. During these cycles, affected individuals usually have no signs or symptoms associated with the condition. However, if treatment is not performed to help prevent morbidity and complications, accumulation of protein deposits (amyloidosis) may occur in the organs and tissues of the body, especially in the kidneys, which can lead to renal failure.

[0005] Chronic inflammation and aberrant immune activity underlie and drive many serious human diseases, ranging from rare and acute inflammatory diseases, rheumatological indications, cardiovascular and metabolic diseases, neurodegenerative diseases, and cancer. In these cases, molecular danger signals generated by dying cells, metabolic disorders, environmental toxins, or diet can act as stimuli that activate the NLRP3 inflammasome (Mangan MSJ et al., Targeting the NLRP3 inflammasome ininflammatory diseases. Nat Rev Drug Discov; 2018; 17(8): 588-606). Once activated, NLRP3 nuclear assembles the inflammasome complex, which coordinates innate and adaptive immune responses to drive a robust inflammatory response (Evavold et al., How Inflammasomes Inform Adaptive Immunity. J. Mol. Biol; 2018; 430(2): 217-237). Inflammasomes are large cytoplasmic multimeric protein complexes that assemble in response to danger signals, and inflammasome activation leads to caspase-1-mediated production of interleukin-1β (IL-1β) and interleukin-18 (IL-18) and pyroptosis (inflammation-mediated cell death) (Dinarello, A clinical perspective of IL-1β as the gatekeeper of inflammation. Eur J Immunol; 2011; 41(5): 1203-17). Through the production of IL-1β and IL-18, the NLRP3 inflammasome has been implicated as a major driver of inflammation associated with autoinflammatory, acute, and chronic inflammatory diseases (e.g., CAPS).In these and other cases, the use of NLRP3 inhibitors to inhibit NLRP3 and thereby block IL-1β, IL-18, and pyroptosis could potentially provide treatment for conditions where persistent inflammasome activation leads to pathology (Ridker PM et al., Modulation of the interleukin-6 signaling pathway and incidence rates of atherosclerotic events and all-cause mortality: analyses from the Canakinumab Anti-Inflammatory Thrombosis Outcomes Study (CANTOS). Eur. Heart J; 2018; 39(38): 3499-3507.).

[0006] In autoinflammatory syndromes, NLRP3 inhibitors may address the underlying etiology of the disease by directly inhibiting the NLRP3 inflammasome. MCC950 is a selective small molecule inhibitor of NLRP3 that has shown activity in inhibiting NLRP3 inflammasome activation in vivo in multiple NLRP3-dependent mouse models and in inhibiting IL-1β release in ex vivo samples of peripheral blood mononuclear cells (PBMCs) from individuals with CAPS (Coll et al., A small-molecule inhibitor of the NLRP3 inflammasome for the treatment of inflammatory diseases. Nat. Med; 2015; 21(3): 248-55). However, administration of the NLRP3 inhibitor MCC950 / CRID3 failed to rescue the CAPS mouse model carrying the L351P mutation in NLRP3, which corresponds to the L353P mutation in human NLRP3 (Vande et al., MCC950 / CRID3 potently targets the NACHT domain of wild-type NLRP3 but not disease-associated mutants for inflammasome inhibition. PLoS Biol; 2019; 17(9): e3000354).

[0007] While supportive care is widely used for milder symptoms, such as fever, rash, and pain, people with autoinflammatory syndromes may require corticosteroids and, in some cases, biologic IL-1 blockers to treat or prevent more severe inflammatory activity. NLRP3 inhibitors may offer additional benefits over biologics to patients and those whose disease is not adequately treated with currently available therapies.

[0008] Provided herein are NLRP3 inhibitors that can be used to prevent or reduce NLRP3 inflammasome responses, thereby addressing unmet medical needs, including the treatment of autoinflammatory syndromes including CAPS, MWS, FCAS, NOMID / CINCA, and FMF. Summary of the invention

[0009] Described herein are methods of treating a subject using NLRP3 inhibitors, particularly Compound I, for use in treating autoinflammatory syndromes. Also described herein are methods of treating autoinflammatory syndromes by administering a therapeutically effective amount of a NLRP3 inhibitor, particularly Compound I, to a subject in need thereof.

[0010] Further provided herein are specific dosage regimens for the methods or uses of the NLRP3 inhibitors described herein, particularly Compound 1.

[0011] In addition, described herein are pharmaceutical combinations for use in treating autoinflammatory syndromes, comprising a) Compound I and b) at least one additional therapeutic agent (optionally in the presence of a pharmaceutically acceptable carrier); and pharmaceutical compositions comprising the same. Preferably, Compound I is Compound IA.

[0012] Other features and advantages of the described methods and uses will become apparent from the following detailed description. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 .An overview diagram of the treatment regimen detailed in Example 1.

[0014] Figure 2 : Overview diagram of the study design for the first-in-human (FIH) study as detailed in Example 2. DETAILED DESCRIPTION

[0015] Methods for treating autoinflammatory syndromes by administering an effective amount of Compound I or a pharmaceutically acceptable salt thereof to a subject in need are described herein. Thus, in one aspect, a method for treating autoinflammatory syndromes is provided, comprising administering an effective amount of Compound I or a pharmaceutically acceptable salt thereof to a subject in need thereof. Compound I for use in treating autoinflammatory syndromes is also provided. In one embodiment of any method or use described herein, the Compound I is Compound IA.

[0016] definition

[0017] In order to make this document more easily understood, certain terms are first defined. Other definitions are set forth throughout this document.

[0018] All patents, published patent applications, publications, references, and other materials mentioned herein are incorporated by reference in their entirety for the purposes described.

[0019] As used herein, the term "comprising" encompasses "including" as well as "consisting of," for example, a composition "comprising" X may consist of X alone, or may include something else, such as X+Y.

[0020] As used herein, the articles “a” and “an” refer to one or to more than one (eg, at least one) of the grammatical object of the article.

[0021] The term "or" is used herein to mean, and is used interchangeably with, the term "and / or," unless the context clearly indicates otherwise.

[0022] The term "about" associated with a reference value as used herein and its grammatical equivalents may include the value itself as well as a range of values ​​that are plus or minus 10% from the value. For example, the amount "about 10" includes 10 and any amount from 9 to 11. For example, the term "about" associated with a reference value may also include a range of values ​​that are plus or minus 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% from the value. In some cases, even if the term "about" is not specifically mentioned, the value described throughout the text may also be "about" the value.

[0023] As used herein, the term "baseline" refers to the degree of a subject's state or condition (e.g., disease) observed prior to treatment according to the described methods and uses (e.g., prior to administration of a compound, e.g., prior to administration of Compound 1, optionally in combination with at least one additional therapeutic agent), or one or more parameters associated with a patient's state.

[0024] As used herein, the term "administering" in relation to a compound (e.g., Compound 1, optionally in combination with at least one additional therapeutic agent) is used to refer to delivery of the compound by any route of delivery. Such delivery may be, for example, intravenous administration or oral administration. Such delivery may also be, for example, subcutaneous administration.

[0025] As used herein, the word "substantially" does not exclude "completely", for example, a composition "substantially free" of Y may be completely free of Y. If necessary, the word "substantially" may be omitted from the definition.

[0026] As used herein, the term "pharmaceutically acceptable" refers to a non-toxic material that does not substantially interfere with the effectiveness of the biological activity of one or more active ingredients.

[0027] As used herein, the term "patient" is used interchangeably with the term "subject" and includes any human or non-human animal. The term "non-human animal" includes all vertebrates, e.g., mammals and non-mammals, such as non-human primates, sheep, dogs, cats, horses, cows, chickens, amphibians, reptiles, etc. In specific embodiments, the compositions, methods, and uses described herein are with respect to human patients or human subjects.

[0028] As used herein, a subject is "in need of" a treatment if the subject has a condition (ie, disease, disorder or syndrome) of interest and would derive a biological, medical, or quality of life benefit from such treatment.

[0029] As used herein, the term "autoinflammatory syndrome" is a form of inflammatory response syndrome that can be triggered or manifested by a variety of factors, such as fatigue, stress or exertion (e.g., FMF), and cold exposure (e.g., CAPS, MWS, FCAS, NOMID / CINCA).

[0030] The terms "treat," "treating," "treatment," "prevent," "preventing," and "prevention" include therapeutic treatments, prophylactic treatments, and uses in which a subject's risk of developing a disorder or other risk factors is reduced. Treatment does not require a complete cure of the disorder, and encompasses alleviation of symptoms or underlying risk factors.

[0031] The term "treating" includes the administration of a compound (e.g., Compound I), optionally in combination with at least one additional therapeutic agent, to prevent the symptoms, complications, or biochemical markers of a disease, condition, disorder, or syndrome (e.g., cryptopyrin-associated periodic syndrome (CAPS), familial cold autoinflammatory syndrome (FCAS), Muckler-Wells syndrome (MWS), neonatal-onset multisystem inflammatory disease / chronic infantile neurocutaneous and articular syndrome (NOMID / CINCA), or familial Mediterranean fever (FMF)), or to delay the onset of the disease, condition, disorder, or syndrome, thereby preventing an outbreak, alleviating the symptoms, or arresting or inhibiting its further development or manifestation.

[0032] As used herein, the terms "prevent," "preventing," or "prevention" in relation to a disease, condition, disorder, or syndrome (e.g., cryptopyrin-associated periodic syndrome (CAPS), familial cold autoinflammatory syndrome (FCAS), Muckler-Wells syndrome (MWS), neonatal-onset multisystem inflammatory disease / chronic infantile neurocutaneous and articular syndrome (NOMID / CINCA), or familial Mediterranean fever (FMF)) refer to the prophylactic treatment of a subject at risk of developing a condition such that the subject has a reduced probability of developing the condition (e.g., a specific disease or disorder associated with CAPS, FCAS, MWS, NOMID / CINCA, or FMF, or its clinical symptoms, such as skin disease, arthralgia, myalgia, headache / migraine, conjunctivitis, fatigue / malaise, and organ or tissue damage).

[0033] For example, "treating familial cold autoinflammatory syndrome (FCAS)" may refer to alleviating, alleviating or modulating at least one symptom or pathological feature associated with familial cold autoinflammatory syndrome (FCAS); for example, low-grade fever, widespread rash, conjunctivitis, polyarthralgia, headache, myalgia and fatigue; for example, it may refer to slowing the progression, thereby reducing or stopping at least one symptom or pathological feature associated with familial cold autoinflammatory syndrome (FCAS); for example, low-grade fever, widespread rash, conjunctivitis, polyarthralgia, headache, myalgia and fatigue. It may also refer to preventing or delaying one or more of the described symptoms, such as slowing the progression of a disease, condition, disorder, manifestation or syndrome progression, stopping or reversing the progression of a disease, condition, disorder, manifestation or syndrome, and improving clinical outcomes.

[0034] "Treatment" may also refer to slowing the progression of, halting or reversing the progression of, and improving clinical outcomes, such as moving from a higher number to a lower number on a 5-point disease-related clinical signs and symptoms scale as follows:

[0035]

[0036]

[0037] As used herein, the term "excipient" or "pharmaceutically acceptable excipient" means a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, carrier, solvent, or encapsulating material. In one embodiment, each component is "pharmaceutically acceptable" in the sense that it is compatible with the other ingredients of the pharmaceutical formulation and is suitable for contact with the tissues or organs of humans and animals without excessive toxicity, irritation, allergic response, immunogenicity or other problems or complications (commensurate with a reasonable benefit / risk ratio). See, e.g., Remington: The Science and Practice of Pharmacy, 21st ed.; Lippincott Williams & Wilkins: Philadelphia, PA, 2005; Handbook of Pharmaceutical Excipients, 6th ed.; Rowe et al., eds.; The Pharmaceutical Press and the American Pharmaceutical Association: 2009; Handbook of Pharmaceutical Additives, 3rd ed.; Ash and Ash, eds.; Gower Publishing Company: 2007; Pharmaceutical Preformulation and Formulation, 2nd ed.; Gibson, ed.; CRC Press LLC: Boca Raton, FL, 2009.

[0038] As used herein, the term "NLRP3 inhibitor" is a compound that inhibits the ability of NLRP3 to induce the production of IL-1β and / or IL-18 by directly binding to NLRP3 or by inactivating, destabilizing, altering the distribution of NLRP3 or otherwise. Typically, the hTHP-1 IC of the NLRP3 inhibitor is 1.0447 / L in a hTHP-1 assay containing 2% fetal bovine serum as defined herein. 50 <1 μM.

[0039] Preferably, the NLRP3 inhibitor is a compound of Compound I, Compound IA or Compound IB. More preferably, the NLRP3 inhibitor is Compound IA.

[0040] As used herein, "compound of Formula I" or "Compound I" are used interchangeably and refer to compounds having the structure shown below and can be synthesized using procedures known in the art and described in WO 2019 / 023147 (incorporated by reference in its entirety).

[0041]

[0042] Compound I, Compound IA or Compound IB can be used in crystalline or amorphous form, as a solvate (eg, hydrate) or in unsolvated form.

[0043]

[0044] Tautomers:

[0045] The scope of the compounds disclosed herein includes tautomeric forms of the compounds. Thus, by way of example, a compound represented by

[0046]

[0047] It is also intended to include tautomeric forms containing the moiety

[0048]

[0049] Stereoisomers:

[0050] Non-limiting exemplary compounds of the formula described herein include stereo sulfur atoms. The present disclosure provides examples of stereoisomer mixtures (e.g., racemic mixtures of enantiomers; mixtures of diastereomers). The present disclosure also describes and illustrates methods for separating individual components of the stereoisomer mixture (e.g., resolving enantiomers of a racemic mixture). For example, compound I represents the following: a non-racemic mixture of compound IA and compound IB, a racemic mixture of compound IA and compound IB; compound IA in enantiomerically pure form; or compound IB in enantiomerically pure form. As used herein, "compound I" is also intended to include compound IA or compound IB in enantiomeric excess. For example, compound IA may be present in an enantiomeric excess of about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or about 99.5%. Alternatively, Compound IB may be present in an enantiomeric excess of about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or about 99.5%.

[0051] Any chemical formula given herein is also intended to represent unlabeled forms of the compound as well as isotopically labeled forms. An isotopically labeled compound has a structure depicted by a formula given herein, but with one or more atoms replaced by an atom having a selected atomic mass or mass number. Isotopes that can be incorporated into the compounds of the present disclosure include, for example, isotopes of hydrogen, carbon, nitrogen, and oxygen, such as 3 H. 11 C. 13 C. 14 C and 15 N. Thus, it will be appreciated that the methods of the invention can or may involve the incorporation of one or more of any of the above isotopes (including, for example, radioactive isotopes (such as 3 H and 14 C)) compounds or the presence of non-radioactive isotopes (such as 2 H and 13 C). This isotope-labeled compound can be used for metabolic studies (using 14 C) reaction kinetics studies (e.g. 2 H or 3 H), detection or imaging techniques, such as positron emission tomography (PET) or single photon emission computed tomography (SPECT), including drug or substrate tissue distribution determinations, or for radiotherapy of patients. Isotope-labeled compounds can generally be prepared by conventional techniques known to those skilled in the art, such as using appropriate isotope-labeled reagents to replace previously employed unlabeled reagents.

[0052] The present invention encompasses the following embodiments, which include all pharmaceutically acceptable salts of compounds useful according to the present invention provided herein. As used herein, "pharmaceutically acceptable salts" refer to derivatives of disclosed compounds, in which the parent compound is modified by converting an existing acid or base moiety into its salt form. Examples of pharmaceutically acceptable salts include, but are not limited to, basic residues such as inorganic or organic acid salts of amines; acidic residues such as alkali metal salts or organic salts of carboxylic acids; etc. Pharmaceutically acceptable salts include, for example, conventional non-toxic salts of the parent compound formed from non-toxic inorganic or organic acids. Pharmaceutically acceptable salts can be synthesized from parent compounds containing basic or acidic moieties by conventional chemical methods. Typically, such salts can be prepared by reacting the free acid or base form of these compounds with a stoichiometric amount of an appropriate base or acid in water or in an organic solvent or in a mixture of the two; typically, non-aqueous media (like ether, ethyl acetate, ethanol, isopropanol or acetonitrile) are preferred. Lists of suitable salts are found in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418 and Journal of Pharmaceutical Science, 66, 2 (1977), each of which is incorporated herein by reference in its entirety. For example, preferred pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic residues such as amines. For example, the salt may be a hydrochloride.

[0053] The phrase "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0054] Unless otherwise indicated, as used herein, the "dose" or amount of an NLRP3 inhibitor (e.g., Compound I) refers to the amount of the free base or free acid form of the compound. For salt forms of the NLRP3 inhibitor, the actual amount will be adjusted based on the salt form used.

[0055] "Effective amount" refers to an amount sufficient to achieve a beneficial or desired result. For example, a therapeutic amount is an amount that achieves a desired therapeutic effect. This amount may be the same or different from a preventive effective amount, which is the amount required to prevent the onset of a disease, condition, disorder or syndrome or related symptoms. The effective amount may be administered in one or more applications, applications or one or more doses. The "therapeutically effective amount" (i.e., effective dose) of a therapeutic compound depends on the selected therapeutic compound. The composition may be administered once or more per day to once or more per week, and also includes less frequent administration, for example, as described herein. It will be appreciated by those skilled in the art that certain factors may affect the dose and timing required for the effective treatment of a subject, including but not limited to the severity of the disease, condition, disorder or syndrome, previous treatment, the general health and / or age of the subject, and other diseases, conditions, disorders or syndromes that exist simultaneously. In addition, treatment of a subject with a therapeutic compound described herein in a therapeutically effective amount may include a single treatment or a series of treatments.

[0056] As used herein, the term "therapeutically effective amount" of a compound described herein refers to an amount of a compound that will elicit a biological or medical response in a subject, such as relieving symptoms, alleviating a condition, slowing or delaying disease progression, or preventing a disease, condition, disorder, manifestation or syndrome, etc. In one non-limiting example, the term "therapeutically effective amount" refers to an amount of a compound described herein that, when administered to a subject, is effective in at least partially alleviating, inhibiting, preventing and / or alleviating an autoinflammatory syndrome (e.g., cryopyrin-associated periodic syndrome (CAPS), familial cold autoinflammatory syndrome (FCAS), Muckler-Wells syndrome (MWS), neonatal-onset multisystem inflammatory disease / chronic infantile neurocutaneous and articular syndrome (NOMID / CINCA), or familial Mediterranean fever (FMF)).

[0057] As defined herein, "combination" refers to a fixed combination in one unit dosage form (e.g., capsule, tablet, sachet or vial), a free (i.e., non-fixed) combination, or a kit of parts for combined administration, in which Compound I and one or more additional therapeutic agents can be administered separately, either independently at the same time or within time intervals, especially when these time intervals allow the combination partners to exhibit a cooperative effect (e.g., a synergistic effect).

[0058] As used herein, the terms "co-administration" or "combined administration" and the like are intended to encompass the administration of an additional therapeutic agent to a single subject (e.g., a subject) in need thereof, and the additional therapeutic agent is intended to include treatment regimens that do not require administration of Compound I and the additional therapeutic agent by the same route of administration and / or at the same time. Each component of the combination described herein can be administered simultaneously or sequentially in any order. Co-administration includes simultaneous, sequential, overlapping, intermittent and / or continuous administration and any combination thereof.

[0059] As used herein, the term "pharmaceutical combination" means a pharmaceutical composition resulting from the combination (eg, admixture) of more than one active ingredient, and includes both fixed and free combinations of the active ingredients.

[0060] The term "fixed combination" means that the active ingredients are administered to a subject simultaneously in the form of a single entity or dosage.

[0061] The term "free combination" (non-fixed combination) means that the active ingredients as defined herein are administered to a subject as different entities simultaneously, concurrently or sequentially without specific time limits, and in any order, wherein such administration provides therapeutically effective levels of the compounds in the subject. In particular, reference to a combination comprising a) Compound I, and b) at least one additional therapeutic agent as used herein (e.g., in any of the Examples or any of the Claims herein) refers to a "non-fixed combination" and can be administered separately, independently at the same time or within time intervals.

[0062] "Simultaneous administration" means administration of the active ingredients as defined herein on the same day. The active ingredients may be administered at the same time (for a fixed or free combination) or one at a time (for a free combination).

[0063] The term "sequential administration" may mean that during a period of consecutive co-administration of two or more days, only one of the active ingredients as defined herein is administered on any given day.

[0064] "Overlapping administration" means that during a period of two or more consecutive days of co-administration, at least one day is for simultaneous administration and at least one day only one of the active ingredients as defined herein is administered.

[0065] "Continuous administration" means a period of co-administration without any blank days. As described above, continuous administration can be simultaneous, sequential or overlapping.

[0066] The term "dose" refers to a specified amount of a drug administered at one time. For example, the dose may be stated on the product packaging or in a product information leaflet.

[0067] As used herein, the term "NLRP3" is intended to include, but is not limited to, nucleic acids, polynucleotides, oligonucleotides, sense and antisense polynucleotide chains, complementary sequences, peptides, polypeptides, proteins, homologous and / or orthologous NLRP3 molecules, isoforms, precursors, mutants, variants, derivatives, splice variants, alleles, different species, and active fragments thereof.

[0068] Examples listed (Examples 1.1 to 1.32):

[0069] 1.1 A NLRP3 inhibitor for use in treating an autoinflammatory syndrome in a subject in need thereof.

[0070] 1.2 A NLRP3 inhibitor for use as described in Example 1.1, wherein the NLRP3 inhibitor is administered to the subject in a total daily dose of about 50 mg to about 500 mg, preferably about 50 mg to about 200 mg, in a single dose or divided doses.

[0071] 1.3 A NLRP3 inhibitor for use as described in Example 1.2, wherein the NLRP3 inhibitor is administered to the subject in a single dose or divided doses at a total daily dose of about 100 mg or about 200 mg.

[0072] 1.4 A NLRP3 inhibitor for use as described in Example 1.3, wherein the NLRP3 inhibitor is administered to the subject at a dose of about 100 mg twice a day for three consecutive days, and about 100 mg is administered once in the morning of the fourth day.

[0073] 1.5 A NLRP3 inhibitor for use as described in any one of Embodiments 1.1 to 1.4, wherein the autoinflammatory syndrome is CAPS, FCAS, MWS, NOMID / CINCA or FMF.

[0074] 1.6 A NLRP3 inhibitor for use as described in any one of Embodiments 1.1 to 1.5, wherein the autoinflammatory syndrome is FCAS.

[0075] 1.7 An NLRP3 inhibitor for use as described in Example 1.6, wherein after administration of the NLRP3 inhibitor, the patient's white blood cell count (WCC) does not increase by more than about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%.

[0076] 1.8 A NLRP3 inhibitor for use as described in Example 1.6, wherein after administration of the NLRP3 inhibitor, the patient exhibits a lower score of at least 1, at least 2, at least 3 on a scale of 1-10 on a Physician's Gross Assessment Scale after cold exposure.

[0077] 1.9 A NLRP3 inhibitor for use as described in Example 1.6, wherein after administration of the NLRP3 inhibitor, the patient exhibits at least a 10%, at least a 20%, at least a 30% lower score on the Physician's Gross Assessment Scale on a scale of 1-100 after cold exposure.

[0078] 1.10 An NLRP3 inhibitor for use as described in any one of Embodiments 1.1 to 1.9, wherein after administration of the NLRP3 inhibitor, the patient's C-reactive protein does not increase by more than about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%.

[0079] 1.11 An NLRP3 inhibitor for use as described in any one of Embodiments 1.1 to 1.10, wherein after administration of the NLRP3 inhibitor, the patient does not exhibit an increase in IL-1β or IL-18 by more than about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100%.

[0080] 1.12 The NLRP3 inhibitor for use as described in any of the preceding enumerated embodiments, wherein the NLRP3 inhibitor is administered orally to the subject.

[0081] 1.13 A NLRP3 inhibitor for use as described in any of the preceding Enumerated Embodiments, wherein the NLRP3 inhibitor is in a tablet formulation.

[0082] 1.14 A NLRP3 inhibitor for use as described in any one of Enumerated Embodiments 1.1 to 1.6, wherein the NLRP3 inhibitor is Compound 1 or a pharmaceutically acceptable salt thereof:

[0083]

[0084] 1.15 A NLRP3 inhibitor for use as described in Embodiment 1.14, wherein Compound I comprises Compound IA enantiomer or a pharmaceutically acceptable salt thereof:

[0085]

[0086] 1.16 A NLRP3 inhibitor for use as described in Embodiment 1.15, wherein Compound I comprises an enantiomer of Compound IB or a pharmaceutically acceptable salt thereof:

[0087]

[0088] 1.17 A NLRP3 inhibitor for use as described in Example 1.15, wherein Compound IA has an enantiomeric excess of at least 90%.

[0089] 1.18 A NLRP3 inhibitor for use as described in Example 1.16, wherein Compound IB has an enantiomeric excess of at least 90%.

[0090] 1.19 A pharmaceutical composition comprising a NLRP3 inhibitor as described in Examples 1.12 to 1.18, for use as described in any of the preceding listed Examples.

[0091] 1.20 A pharmaceutical combination comprising a NLRP3 inhibitor as described in Embodiments 1.12 to 1.18 and at least one additional therapeutic agent, for use as described in any of the preceding enumerated embodiments, further comprising.

[0092] Uses and methods

[0093] The following and other places in this document include multiple embodiments of the methods and uses described herein. It should be appreciated that the features specified in each embodiment can be combined with other specified features to provide further embodiments:

[0094] The following examples teach the use of any NLRP3 inhibitor, and are not limited to Compound I. Preferably, Compound I of the following examples is an enantiomeric excess of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 99.5% of Compound IA (i.e., R enantiomer). Preferably, Compound IA has an enantiomeric excess of at least 90%. More preferably, Compound IA has an enantiomeric excess of at least 95%.

[0095] Example 1

[0096] In one embodiment, provided herein is a method for treating an autoinflammatory syndrome in a subject in need thereof or alleviating its symptoms, the method comprising administering an effective amount of Compound 1 or a pharmaceutically acceptable salt thereof. In one embodiment, provided herein is Compound 1 or a pharmaceutically acceptable salt thereof for use in treating an autoinflammatory syndrome in a subject in need thereof. In some embodiments, provided herein is the use of Compound 1 for the manufacture of a medicament for treating an autoinflammatory syndrome.

[0097] Example 2

[0098] In one embodiment, provided herein is a method for treating a subject in need of cryopyrin-associated periodic syndrome (CAPS) or alleviating its symptoms, the method comprising administering an effective amount of Compound 1 or a pharmaceutically acceptable salt thereof. In one embodiment, provided herein is Compound 1 or a pharmaceutically acceptable salt thereof for use in treating a subject in need of cryopyrin-associated periodic syndrome (CAPS). In some embodiments, provided herein is the use of Compound 1 for the manufacture of a medicament for treating cryopyrin-associated periodic syndrome (CAPS).

[0099] Example 3

[0100] In one embodiment, provided herein is a method for treating familial cold autoinflammatory syndrome (FCAS) in a subject in need thereof or alleviating its symptoms, the method comprising administering an effective amount of Compound 1 or a pharmaceutically acceptable salt thereof. In one embodiment, provided herein is a Compound 1 or a pharmaceutically acceptable salt thereof for use in treating familial cold autoinflammatory syndrome (FCAS) in a subject in need thereof. In some embodiments, provided herein is the use of Compound 1 for the manufacture of a medicament for treating familial cold autoinflammatory syndrome (FCAS).

[0101] Example 4

[0102] In one embodiment, provided herein is a method for treating Muckler-Wells syndrome (MWS) in a subject in need thereof or alleviating its symptoms, the method comprising administering an effective amount of Compound 1 or a pharmaceutically acceptable salt thereof. In one embodiment, provided herein is Compound 1 or a pharmaceutically acceptable salt thereof for use in treating Muckler-Wells syndrome (MWS) in a subject in need thereof. In some embodiments, provided herein is the use of Compound 1 for the manufacture of a medicament for treating Muckler-Wells syndrome (MWS).

[0103] Example 5

[0104] In one embodiment, provided herein is a method for treating a neonatal-onset multisystem inflammatory disease / chronic infantile neurocutaneous and articular syndrome (NOMID / CINCA) in a subject in need thereof or alleviating its symptoms, the method comprising administering an effective amount of Compound 1 or a pharmaceutically acceptable salt thereof. In one embodiment, provided herein is Compound 1 or a pharmaceutically acceptable salt thereof for use in treating a neonatal-onset multisystem inflammatory disease / chronic infantile neurocutaneous and articular syndrome (NOMID / CINCA) in a subject in need thereof. In some embodiments, provided herein is the use of Compound 1 for the manufacture of a medicament for treating a neonatal-onset multisystem inflammatory disease / chronic infantile neurocutaneous and articular syndrome (NOMID / CINCA).

[0105] Example 6

[0106] In one embodiment, provided herein is a method for treating familial Mediterranean fever (FMF) in a subject in need thereof or alleviating its symptoms, the method comprising administering an effective amount of Compound 1 or a pharmaceutically acceptable salt thereof. In one embodiment, provided herein is Compound 1 or a pharmaceutically acceptable salt thereof for use in treating familial Mediterranean fever (FMF) in a subject in need thereof. In some embodiments, provided herein is the use of Compound 1 for the manufacture of a medicament for treating familial Mediterranean fever (FMF).

[0107] In any embodiments described herein, the patient's white blood cell count (WCC) does not increase by more than about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100% after administration of the NLRP3 inhibitor.

[0108] In any embodiments described herein, the patient's C-reactive protein does not increase by more than about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100% after administration of the NLRP3 inhibitor.

[0109] In any embodiments described herein, the patient does not exhibit more than about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100% increase in IL-1β or IL-18 following administration of the NLRP3 inhibitor.

[0110] In any embodiments of treating or reducing a symptom of FCAS described herein, after administration of the NLRP3 inhibitor, the patient exhibits at least 1, at least 2, at least 3 lower scores on the Physician's Gross Assessment Scale on a scale of 1-10 after cold exposure. In any embodiments of treating or reducing a symptom of FCAS described herein, after administration of the NLRP3 inhibitor, the patient exhibits at least 10%, at least 20%, at least 30% lower scores on the Physician's Gross Assessment Scale on a scale of 1-100 after cold exposure.

[0111] In any embodiment described herein, Compound 1 or a pharmaceutically acceptable salt thereof may be administered to a subject at a total daily dose (as measured as a non-salt equivalent) of about 50 mg to about 200 mg in a single dose or divided doses. In a specific embodiment, Compound 1 is administered to a subject at a total daily dose of about 200 mg in a single dose or divided doses. In yet a specific embodiment, Compound 1 is administered to a subject at a dose of about 100 mg, twice a day for three consecutive days, and once in the morning on the fourth day at about 100 mg.

[0112] In some embodiments, a pharmaceutical composition is provided herein, comprising Compound I or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient. In a specific embodiment, the pharmaceutical composition is a tablet. In another specific embodiment, the pharmaceutical composition is administered as a whole or crushed tablet. In some embodiments, in each unit dose, the pharmaceutical composition includes about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, or about 100 mg.

[0113] Provided herein is a pharmaceutical composition comprising Compound 1 or a pharmaceutically acceptable salt thereof for use in any of the embodiments described herein.

[0114] In any of the embodiments described herein, Compound 1 or a pharmaceutically acceptable salt thereof is orally administered to a subject in need thereof. In some embodiments, Compound 1 is in the form of a tablet, which is administered whole or subdivided (i.e., crushed prior to administration). In certain embodiments, Compound 1 may be administered via a nasogastric tube, for example when the patient is unable to swallow.

[0115] For all embodiments defined herein, the preferred NLRP3 inhibitor is Compound IA. In specific embodiments, Compound IA is in an enantiomeric excess of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5%.

[0116] Subjects

[0117] As described herein, a subject receiving the currently described NLRP3 inhibitors may develop symptoms of or be at risk for developing familial cold autoinflammatory syndrome (FCAS), such as described above.

[0118] Combination therapy

[0119] In practicing some of the methods of treatment or uses described herein, a therapeutically effective amount of Compound I is administered to a patient such as a mammal (e.g., a human). In addition, for less severe cases of FCAS, patients can be treated in a supportive manner to control symptoms such as fever, muscle pain, or fatigue. For more severe cases of FCAS, immunosuppressants such as corticosteroids may be required, but judgment must be made to avoid negating the effects of drugs designed to activate the immune system.

[0120] Pharmaceutical composition

[0121] When combined with a pharmaceutically acceptable carrier, Compound I can be used as a pharmaceutical composition. In addition to Compound I, such compositions may also contain carriers, various diluents, fillers, salts, buffers, stabilizers, solubilizers, and other known materials. The characteristics of the carrier will depend on the route of administration. The pharmaceutical compositions used for the compositions, uses, and methods described herein may also contain at least one or more additional therapeutic agents for treating a specific targeted disorder, disease, condition, or syndrome. Such additional factors and / or agents may be included in the pharmaceutical composition to produce a synergistic effect with Compound I.

[0122] In a specific embodiment, Compound I can be administered in combination with one or more conventional pharmaceutical excipients. Pharmaceutically acceptable excipients include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, self-emulsifying drug delivery systems (SEDDS) (such as d-α-tocopherol polyethylene glycol 1000 succinate), surfactants used in pharmaceutical dosage forms (such as Tween, poloxamer or other similar polymer delivery matrices), serum proteins (such as human serum albumin), buffer substances (such as phosphates, tris, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes (such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts)), colloidal silicon dioxide, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, waxes, polyethylene-polyoxypropylene block polymers, and lanolin. Cyclodextrins such as α-, β- and γ-cyclodextrins, or chemically modified derivatives such as hydroxyalkylcyclodextrins (including 2- and 3-hydroxypropyl-β-cyclodextrins), or other dissolved derivatives can also be used to enhance the delivery of the compounds described herein. Dosage forms or compositions containing a chemical entity as described herein in the range of 0.005% to 100% can be prepared, with the balance being made up by non-toxic excipients. The intended composition may contain 0.001%-100%, 0.1%-95% in one embodiment, 75%-85% in another embodiment, and 20%-80% of the chemical entity provided herein in another embodiment. The actual methods of preparing such dosage forms are known or will be apparent to those skilled in the art; for example, see Remington: The Science and Practice of Pharmacy, 22nd Edition (Pharmaceutical Press, London, UK. 2012).

[0123] Routes of administration and composition components

[0124] In some embodiments, the chemical entities described herein, or pharmaceutical compositions thereof, may be administered to a subject in need thereof by any accepted route of administration. Acceptable routes of administration include, but are not limited to, buccal, dermal, intracervical, endosinusial, intratracheal, enteral, epidural, interstitial, intraperitoneal, intraarterial, intrabronchial, intracapsular, intracerebral, intracisternal, intracoronary, intradermal, intraductal, intraduodenal, intradural, intraepidermal, intraesophageal, intragastric, intragingival, intraileal, intralymphatic, intramedullary, intrameningeal, intramuscular, intraovarian, intraperitoneal, intraprostatic, intrapulmonary, intrasinal, intraspinal, intrasynovial, intratesticular, intrathecal, intratubular, intratumoral, intrauterine, intravascular, intravenous, nasal, nasogastric, oral, parenteral, transdermal, epidural, rectal, respiratory (inhalation), subcutaneous, sublingual, submucosal, topical, transdermal, transmucosal, transtracheal, ureteral, urethral and vaginal administration. In certain embodiments, the preferred route of administration is parenteral (e.g., intratumoral).

[0125] The composition can be formulated for parenteral administration, e.g., for injection via intravenous, intramuscular, subcutaneous, or even intraperitoneal routes. Typically, such compositions can be prepared as injectables that are liquid solutions or suspensions; solid forms suitable for preparing solutions or suspensions when liquids are added prior to injection can also be prepared; and the formulations can also be emulsified. The preparation of such formulations will be known to those skilled in the art in light of the present disclosure.

[0126] Pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions; formulations including sesame oil, peanut oil, or aqueous propylene glycol; and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, the form must be sterile and must have fluidity to the extent that it can be easily syred. It should also be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi.

[0127] The carrier can also be a solvent or dispersion medium containing, for example, water, ethanol, polyols (for example, glycerol, propylene glycol and liquid polyethylene glycol, etc.), suitable mixtures thereof and vegetable oils. Suitable fluidity can be maintained, for example, in the following manner: by using a coating (such as lecithin), by maintaining the required particle size in the case of a dispersion, and by using a surfactant. Prevention of microbial action can be achieved by various antibacterial and antifungal agents (for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal (thimerosal) etc.). In many cases, it is preferred to include isotonic agents, such as sugar or sodium chloride. The absorption of the injectable composition can be extended by using an agent (such as aluminum monostearate and gelatin) that delays absorption in the composition.

[0128] Sterile injectable solutions are prepared by mixing the active compound in the required amount with various other ingredients listed above (as required) in an appropriate solvent, followed by filtration sterilization. Typically, dispersions are prepared by mixing various sterilized active ingredients into a sterile vehicle containing an alkaline dispersion medium and other ingredients from those listed above. For the preparation of sterile powders for sterile injectable solutions, preferred preparation methods are vacuum drying and freeze drying techniques, which produce a powder of the active ingredient plus any additional desired ingredients from its previous sterile filtered solution.

[0129] Intratumoral injection is discussed in, for example, Lammers et al., “Effect of Intratumoral Injection on the Biodistribution and the Therapeutic Potential of HPMA Copolymer-Based Drug Delivery Systems,” Neoplasia. 2006, 10, 788-795.

[0130] In certain embodiments, the chemical entities described herein or pharmaceutical compositions thereof are suitable for topical, external administration to the digestive tract or gastrointestinal (GI) tract, e.g., rectal administration. Rectal compositions include, but are not limited to, enemas, rectal gels, rectal foams, rectal aerosols, suppositories, jelly-like suppositories, and enemas (e.g., retention enemas).

[0131] Pharmacologically acceptable excipients that can be used in rectal compositions as gels, creams, enemas, or rectal suppositories include, but are not limited to, any one or more of the following: cocoa butter glycerides, synthetic polymers such as polyvinyl pyrrolidone, PEG (e.g., PEG ointment), glycerin, glycerinated gelatin, hydrogenated vegetable oils, poloxamers, mixtures of polyethylene glycols of various molecular weights and fatty acid esters of polyethylene glycol, petrolatum, anhydrous lanolin, shark liver oil, sodium saccharinate, menthol, sweet almond oil, sorbitol, sodium benzoate, anoxidase, SBN, vanilla essential oil, parabens in aerosol, phenoxyethanol, sodium methylparaben, sodium propylparaben, diethylamine, carbomer, carbopol, methoxybenzoate, cetearyl alcohol, cocoyl capryloyl caprate, isopropyl alcohol, propylene glycol, liquid paraffin, xanthan gum, carboxy-metabisulfite, sodium edetate, sodium benzoate, potassium metabisulfite, grape seed extract, methylsulfonylmethane (MSM), lactic acid, glycine, vitamins (such as vitamins A and E), and potassium acetate.

[0132] In certain embodiments, suppositories can be prepared by mixing the chemical entities described herein with suitable non-irritating excipients or carriers (such as cocoa butter, polyethylene glycol or suppository wax), which are solid at ambient temperature but liquid at body temperature and thus melt in the rectum and release the active compound. In other embodiments, compositions for rectal administration are in the form of enemas.

[0133] In other embodiments, the compounds described herein, or pharmaceutical compositions thereof, are suitable for local delivery to the digestive or gastrointestinal (GI) tract via oral administration (eg, solid or liquid dosage forms).

[0134] Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In such solid dosage forms, the chemical entity is mixed with one or more pharmaceutically acceptable excipients such as sodium citrate or dicalcium phosphate and / or: a) fillers or extenders such as starch, lactose, sucrose, glucose, mannitol and silicic acid, b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and gum arabic, c) humectants such as glycerol, d) disintegrants such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) solution retardants such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as, for example, cetyl alcohol and glyceryl monostearate, h) absorbents such as kaolin and bentonite clays, and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may also contain buffering agents.Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar and high molecular weight polyethylene glycols.

[0135] In one embodiment, the composition is in the form of a unit dosage form, such as a pill or tablet, and thus the composition may contain a diluent, such as lactose, sucrose, dicalcium phosphate, etc., together with the chemical entities provided herein; a lubricant, such as magnesium stearate, etc.; and a binder, such as starch, gum arabic, polyvinyl pyrrolidone, gelatin, cellulose, cellulose derivatives, etc. In another solid dosage form, a powder, a pellet (marume), a solution or a suspension (e.g., in propylene carbonate, vegetable oil, PEG, poloxamer 124, or triglycerides) is encapsulated in a capsule (gelatin or cellulose-based capsule). It is also contemplated that one or more chemical entities or additional active agents provided herein are physically separated unit dosage forms; for example, capsules containing particles of each drug (or tablets in a capsule); double-layer tablets; double-chamber gel caps, etc. Enteric coatings or delayed release oral dosage forms are also contemplated.

[0136] Other physiologically acceptable compounds include wetting agents, emulsifiers, dispersants or preservatives, especially for preventing the growth or action of microorganisms. Various preservatives are well known and include, for example, phenol and ascorbic acid.

[0137] In certain embodiments, the excipient is sterile and generally free of undesirable substances. These compositions can be sterilized by conventional well-known sterilization techniques. For various oral dosage form excipients such as tablets and capsules, sterility is not required. USP / NF standards are generally sufficient.

[0138] In certain embodiments, the solid oral dosage form may further include one or more components that chemically and / or structurally facilitate the delivery of the composition to the stomach or lower GI; for example, the ascending colon and / or transverse colon and / or distal colon and / or small intestine. Exemplary formulation techniques are described, for example, in Filipski, KJ et al., Current Topics in Medicinal Chemistry, 2013, 13, 776-802, which is incorporated herein by reference in its entirety.

[0139] Examples include upper-GI targeting technologies, eg, the Accordion Pill (Intec Pharma), floating capsules, and materials capable of adhering to the mucosal wall.

[0140] Other examples include lower-GI targeting technology. For targeting various regions in the intestinal tract, several enteric / pH reactive coatings and excipients are available. These materials are typically polymers, which are designed to dissolve or erode under a specific pH range selected in the GI region based on the desired drug release. These materials are also used to protect acid-labile drugs from the influence of gastric juice or limit exposure when active ingredients can stimulate the upper GI (for example, hydroxypropyl methylcellulose phthalate series, Coateric (polyvinyl acetate phthalate), cellulose acetate phthalate, hydroxypropyl methylcellulose acetate succinate, Eudragit series (methacrylic acid-methyl methacrylate copolymer) and Marcoat). Other technologies include dosage forms that react to local flora in the gastrointestinal (GI) tract, pressure-controlled colon delivery capsules and Pulsincap.

[0141] Ophthalmic compositions may include, but are not limited to, any one or more of the following: viscogen (e.g., carboxymethylcellulose, glycerol, polyvinylpyrrolidone, polyethylene glycol); stabilizer (e.g., Pluronic (triblock copolymer), cyclodextrin); preservative (e.g., benzalkonium chloride, ETDA, SofZia (boric acid, propylene glycol, sorbitol and zinc chloride; Alcon Laboratories, Inc.), Purite (stabilized oxychlorine complex; Allergan, Inc.).

[0142] Topical compositions can include ointments and creams. Ointments are semisolid preparations, which are typically based on petrolatum or other petroleum derivatives. Creams containing selected active agents are typically viscous liquids or semisolid emulsions, usually oil-in-water or water-in-oil types. Cream bases are typically water-washable and contain an oil phase, an emulsifier, and an aqueous phase. The oil phase (sometimes also referred to as the "inner" phase) is typically composed of petrolatum and fatty alcohols (such as cetyl alcohol or stearyl alcohol); the volume of the aqueous phase is usually (although not necessarily) greater than the oil phase, and usually contains a wetting agent. The emulsifier in the cream formulation is usually a nonionic, anionic, cationic, or amphoteric surfactant. With respect to other carriers or vehicles, the ointment base should be inert, stable, non-irritating, and non-sensitizing.

[0143] In any of the foregoing embodiments, the pharmaceutical compositions described herein may include one or more of the following: lipids, multilamellar vesicles with cross-links between bilayers, biodegradable poly (D, L-lactic-co-glycolic acid) [PLGA]-based or polyanhydride-based nanoparticles or microparticles, and nanoporous particle-supported lipid bilayers.

[0144] Dosage regimen and route of administration

[0145] The dosage regimen is adjusted to provide the optimal desired response (e.g., a therapeutic response). Depending on the compound used, the targeted disease, condition, disorder or syndrome, and their associated stages, the dosage regimen (i.e., the dosage and / or frequency of administration of the pharmaceutical composition comprising Compound I) may vary. Depending on the compound used, the disease, condition, disorder or syndrome, and their associated stages, the dosage regimen (i.e., the dosage and / or frequency of administration of the pharmaceutical combination comprising a) Compound I and b) at least one additional therapeutic agent) may vary.

[0146] For the administration of Compound I in the method for treating autoinflammatory syndrome, the dosage is in the range of about 0.0001 to about 100 mg / kg of subject body weight, and more typically in the range of about 0.01 to about 30 mg / kg of subject body weight. In a specific embodiment, Compound I is administered at a daily dose of about 50 mg to about 500 mg, about 50 mg to about 200 mg, about 50 mg to about 150 mg, about 50 mg to about 100 mg, about 50 mg. In a specific embodiment, Compound I is administered at a daily dose of about 50 mg, about 100 mg, about 150 mg, or about 200 mg. In a specific embodiment, Compound I is administered once a day. In other embodiments, Compound I is administered twice, three times, or four times a day. In a preferred embodiment, Compound I is administered once or in two divided doses at a total daily dose of about 200 mg. Preferably, Compound I is Compound IA. Preferably, Compound IA is administered at a total daily dose of about 50 mg to 500 mg. More preferably, Compound 1A is administered at a daily dose of 200 mg.

[0147] In some embodiments, Compound IA is administered at a daily dose of 200 mg. In some embodiments, 100 mg of Compound IA is administered twice daily.

[0148] In some embodiments, the period of time for administering a compound described herein lasts for 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, or longer. In another embodiment, the period of time for stopping administration lasts for 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, or longer. Doses are repeated as necessary and may range from about once per week to about once every 10 weeks, for example once every 4 weeks or once every 8 weeks.

[0149] Reagent test kit

[0150] Also included herein are kits for use in methods for treating or preventing cytokine release syndrome or cytokine storm syndrome, which may include Compound 1 or a pharmaceutical composition comprising Compound 1 in liquid or lyophilized form. In addition, such kits may include tools (e.g., syringes and vials, prefilled syringes, prefilled pens) and instructions for use for administering Compound 1. These kits may contain, for example, additional therapeutic agents for delivery in combination with Compound 1 (described elsewhere herein).

[0151] The phrase "means for administering" is used to indicate any available instrument for administering a drug systemically to a patient, including but not limited to droppers, prefilled syringes, vials and syringes, injection pens, autoinjectors, intravenous (iv) drip sets and bags, pumps, etc. Using such items, a patient can self-administer the drug (i.e., administer the drug on their own), a caregiver can administer the drug to the patient, or a physician or other medical professional can administer the drug.

[0152] Each component of the kit is typically enclosed in an individual container, and all of the various containers are within a single package with instructions for use.

[0153] It should be understood that each embodiment can be combined with one or more other embodiments until such combination is consistent with the description of the embodiment. It should also be understood that the embodiments provided above should be understood to include all embodiments, including such embodiments generated by the combination of embodiments.

[0154] Other features, objects, and advantages of the described methods and uses will be apparent from the description and drawings, and from the claims.

[0155] Synthesis of Compound I

[0156] Compounds I, IA and IB were synthesized according to the synthesis defined in WO 2019 / 023147 for Examples 4, 5 and 6 and as detailed below. However, the compounds can be assembled in various ways, using the relevant reaction procedures to build the final molecules in a modular manner allowing different reaction orders and / or different reagents.

[0157] The progress of the reaction is usually monitored by TLC or LC-MS. The identity of the product is usually confirmed by LC-MS. LC-MS is recorded using the following method:

[0158] Method A: Shim-pack XR-ODS, C18, 3x 50mm, 2.5um column, 1.0uL injection, 1.5mL / min flow rate, 90-900amu scan range, 190-400nm UV range, 5%-100% (1.1min), gradient from 100% (0.6min) with ACN (0.05% TFA) and water (0.05% TFA), 2 minutes total run time.

[0159] The final target was purified by preparative HPLC. Preparative HPLC was performed using the following method:

[0160] Method B: Preparative HPLC: column, XBridge Shield RP18 OBD (19 x 250 mm, 10 um); mobile phase, water (10 mmol / L NH4HCO3) and ACN, UV detection 254 / 210 nm.

[0161] On BRUKER NMR 300.03MHz, DUL-CH, ULTRASHIELD TM 300,AVANCE II 300B-ACS TM 120 or BRUKER NMR 400.13MHz, BBFO, ULTRASHIELD TM 400,AVANCE III 400,B-ACS TM NMR was recorded on a BRUKER AC120 or BRUKER AC250 NMR instrument with TMS as reference and ppm (parts per million) as the measurement unit.

[0162] Scenario 1:

[0163]

[0164] Compound I:

[0165]

[0166] Compound I: N'-(1,2,3,5,6,7-hexahydro-symmetric indacene-4-ylcarbamoyl)-2-(2-hydroxypropyl)-

[0167] -2-yl)thiazole-5-sulfonylimideamide

[0168] Step 1: N-(tert-butyldimethylsilyl)-N'-(1,2,3,5,6,7-hexahydro-s-indacene-4-ylcarbamoyl)-2-(2-hydroxypropan-2-yl)thiazole-5-sulfonylimide amide

[0169] A solution of N'-(tert-butyldimethylsilyl)-2-(2-hydroxypropyl-2-yl)thiazole-5-sulfonylimide amide (intermediate I) (336mg, 1.0mmol) in THF (10mL) was placed in a 50mL round-bottom flask. NaH (60%wt, 80mg, 2.0mmol) was added to the solution in batches at 0°C. The solution was stirred at 0°C for 15 minutes, and then a solution of 4-isocyanato-1,2,3,5,6,7-hexahydro-symmetric indacene (209mg, 1.1mmol) in THF (5mL) was added dropwise with stirring at RT. The resulting solution was stirred for 12h at RT. The reactant was then quenched by adding 10mL NH4Cl (saturated). The resulting solution was extracted with 3x 10mL DCM and the combined organic layer was concentrated under vacuum. This produces 535mg (crude product) of the title compound in the form of brown oil. MS-ESI:535.0(M+1).

[0170] Step 2: N'-(1,2,3,5,6,7-hexahydro-4-s-indacenecarbamoyl)-2-(2-hydroxypropan-2-yl)thiazole-5-sulfonylimideamide

[0171] Into a 50-mL round-bottom flask was placed a solution of N-(tert-butyldimethylsilyl)-N'-(1,2,3,5,6,7-hexahydro-symmetrical indacene-4-ylcarbamoyl)-2-(2-hydroxypropyl-2-yl)thiazole-5-sulfonylimide amide (535mg, crude product, 1.0mmol) in THF (10mL). HF / Py (70%wt, 143mg, 5.0mmol) was added dropwise to the solution at 0°C. The solution was stirred for 4h at RT. The reaction was then quenched by adding 10mL of water. The resulting solution was extracted with 3x 10mL ethyl acetate and the combined organic layers were concentrated under vacuum. The crude product was purified by preparative HPLC using method B with ACN / water (20% to 60% in 10 minutes). This produced 189mg (45%, 2 steps) of compound I as a white solid.

[0172] Compound I: MS-ESI: 421.0 (M+1). 1 H NMR(400MHz,DMSO-d6)δ8.46(br s,1H),8.04(s,1H),7.80(br s,2H),6.86(s,1H)6.28(s,1H),2.88-2.71(m,4H),2.71-2.56(m,4H),2.02-1.80(m,4H),1.49(s,6H).

[0173] Compound IA and Compound IB:

[0174]

[0175] Compounds IA and IB: ( R) and (S)-N'-(1,2,3,5,6,7-hexahydro-sym-indacene-4-ylcarbamoyl 2-(2-hydroxypropan-2-yl)thiazole-5-sulfonylimideamide

[0176] Step 3: Chiral separation.

[0177] The compound I product (189 mg) obtained as described in the previous step was separated by chiral preparative HPLC using the following conditions: column, CHIRAL Cellulose-SB, 2*25 cm, 5 um; mobile phase, Hex (0.1% DEA) and EtOH (maintaining 20% ​​EtOH for 16 min); flow rate, 20 mL / min; detector, UV 254 / 220 nm. This produced 70 mg of compound IB (front peak, 99% ee) as a white solid and 65 mg of compound IA (second peak, 97.5% ee) as a white solid.

[0178] Compound IB: MS-ESI: 421.0 (M+1). 1 H NMR(400MHz,DMSO-d6)δ8.43(br s,1H),8.05(s,1H),7.83(br s,2H),6.87(s,1H)6.29(s,1H),2.82-2.71(m,4H),2.71-2.56(m,4H),2.02-1.80(m,4H),1.50(s,6H).

[0179] Compound IA: MS-ESI: 421.0 (M+1). 1 H NMR(400MHz,DMSO-d6)δ8.41(br s,1H),8.05(s,1H),7.83(s,2H),6.87(s,1H)6.27(s,1H),2.82-2.71(m,4H),2.71-2.56(m,4H),2.02-1.80(m,4H),1.50(s,6H).

[0180] Intermediate I of Scheme 1 was synthesized according to the synthesis shown in WO 2019 / 023147 and as provided in Scheme 2 below.

[0181] Scenario 2:

[0182]

[0183] Intermediate I

[0184]

[0185] N'-(tert-Butyldimethylsilyl)-2-(2-hydroxypropan-2-yl)thiazole-5-sulfonylimideamide

[0186] Step 1: 2-(2-methyl-1,3-dioxolan-2-yl)thiazole

[0187] Into a 500-mL round-bottom flask was placed a solution of 1-(thiazol-2-yl)ethanone (20 g, 157.0 mmol) in toluene (300 mL) and ethane-1,2-diol (19.5 g, 314 mmol). TsOH (2.7 g, 15.7 mmol) was added to the solution. The resulting solution was refluxed overnight and water was separated from the solution during reflux. The resulting solution was diluted with 200 mL of water and extracted with 2x100 mL of ethyl acetate. The organic layers were combined, dried over anhydrous Na2SO4, and then concentrated under vacuum. This produced 26.6 g (99%) of the title compound as a light yellow oil. MS-ESI: 172.0 (M+1).

[0188] Step 2: 2-(2-methyl-1,3-dioxolan-2-yl)thiazole-5-sulfonamide

[0189] A solution of 2-(2-methyl-1,3-dioxolane-2-yl)thiazole (14g, 81.6mmol) in THF (200mL) was placed in a 500-mL 3-neck round-bottom flask purged with nitrogen and maintained under nitrogen. Then n-BuLi (2.5M, in THF, 35.2mL, 88.0mmol) was added dropwise with stirring at -78°C. The resulting solution was stirred for 0.5h at -78°C, and then SO2 was introduced into the above reaction mixture. The reactant was slowly warmed to RT, and then NCS (12.8g, 95.86mmol) was added. The resulting solution was stirred for 1h at RT. The solid was filtered out. The resulting filtrate was concentrated under vacuum, and then diluted in DCM (160mL). A saturated solution of ammonia in DCM (300mL) was added to the above dilution. The resulting solution was stirred for 3h at RT, and then concentrated under vacuum. The residue was applied to a silica gel column and eluted with a gradient of ethyl acetate / petroleum ether (1:20 to 1:5). This resulted in 12.5 g (61%) of the title compound as a yellow solid. MS-ESI: 251.0 (M+1).

[0190] Step 3: 2-Acetylthiazole-5-sulfonamide

[0191] Into a 250-mL round-bottom flask was placed a solution of 2-(2-methyl-1,3-dioxolane-2-yl)thiazole-5-sulfonamide (12.5 g, 50.0 mmol) in THF (125 mL). To the above solution was added an aqueous HCl solution (4 N, 50.0 mL). The resulting solution was stirred at 70 ° C for 6 h. The resulting solution was diluted with 100 mL of water and extracted with 2 x 200 mL of ethyl acetate. The organic layers were combined, dried over anhydrous Na2SO4, and then concentrated under vacuum. The residue was applied to a silica gel column and eluted with a gradient of ethyl acetate / petroleum ether (1:2 to 1:1). This produced 9.3 g (90%) of the title compound as a yellow solid. MS-ESI: 207.0 (M+1).

[0192] Steps 4-6 used the same procedure as specified for the conversion of compound Z to compound Y shown in Scheme 3 to obtain intermediate I from compound Id. MS-ESI: 336.1 (M+1).

[0193] Scenario 3:

[0194]

[0195] Compound Y

[0196]

[0197] N'-(tert-Butyldimethylsilyl)-5-(2-hydroxypropan-2-yl)thiazole-2-sulfonylimideamide

[0198] Step 1: Methyl 2-mercaptothiazole-5-carboxylate

[0199] Into a 2-L round-bottom flask was placed methyl 2-bromothiazole-5-formate (100 g, 450 mmol), EtOH (1000 mL), sodium bisulfide (50 g, 890 mmol). The resulting solution was stirred at 80 ° C for 2 h and then cooled to 0 ° C in a water / ice bath. The pH value of the solution was adjusted to 3 with hydrogen chloride (1 N). The solid was collected by filtration. This produced 63.2 g (80%) of the title compound as a light yellow solid. MS-ESI: 176.0 (M+1).

[0200] Step 2: Methyl 2-(chlorosulfonyl)thiazole-5-carboxylate

[0201] Into a 1-L round-bottom flask, 2-mercaptothiazole-5-methyl formate (30 g, 170 mmol) and acetic acid (300 mL) were placed. Sodium hypochlorite (300 mL, 8%-10% wt.) was subsequently added in batches at 0 ° C. The resulting solution was stirred for 2 h at RT, and then diluted with 500 mL of water. The solution was extracted with 3 x 300 mL of DCM, and the combined organic layers were washed with 2 x 300 mL of brine and dried over anhydrous Na2SO4. The crude product in a yellow solution in DCM was used for the next step.

[0202] Step 3: Methyl 2-sulfamoylthiazole-5-carboxylate

[0203] Into a 2-L round-bottom flask is placed 2-(chlorosulfonyl)thiazole-5-formic acid methyl ester in a crude solution in DCM (900mL). NH3(g) is introduced into the solution at less than 0°C for 20 minutes. The resulting solution is stirred for 1h at RT, and then concentrated under vacuum. The residue is applied to a silica gel column and eluted with ethyl acetate / petroleum ether (1:5 to 1:3). This produces 23g (75%, 2 steps) of the title compound as a white solid. MS-ESI: 223.0 (M+1).

[0204] Step 4: 5-(2-Hydroxypropan-2-yl)thiazole-2-sulfonamide

[0205] A solution of 2-sulfamoylthiazole-5-formic acid methyl ester (15g, 67.5mmol) in THF (150mL) was placed in a 500-mL round-bottom flask purged with nitrogen and maintained under nitrogen. MeMgBr / THF (3M, 90mL) was then added dropwise with stirring at 0°C. The resulting solution was stirred for 14h at RT and then quenched by adding 100mL NH4Cl (saturated). The resulting solution was extracted with 3x 150mL DCM. The organic layers were combined and dried over anhydrous Na2SO4, then concentrated under vacuum. The residue was applied to a silica gel column and eluted with ethyl acetate / petroleum ether (1:5 to 1:3). This produced 11.5g (78%) of the title compound as a white solid. MS-ESI: 223.0 (M+1), 221.0 (M-1), respectively in positive ion mode and negative ion mode.

[0206] Step 5: N-(tert-Butyldimethylsilyl)-5-(2-hydroxypropan-2-yl)thiazole-2-sulfonamide

[0207] To be purged with nitrogen and maintained in a 250-mL 3-necked round-bottom flask under nitrogen, a solution of 5-(2-hydroxypropyl-2-yl)thiazole-2-sulfonamide (5g, 22.5mmol) in THF (100mL) is placed. Then in an ice / water bath, NaH (60%wt, 1.8g, 45.0mmol) is added in batches to the above solution. After stirring in a water / ice bath for 20 minutes, a solution of TBSCl (4.1g, 27.2mmol) in THF (10mL) is subsequently added dropwise with stirring at 0°C. The resulting solution is stirred for 4h at RT. The reactant is quenched with saturated NH4Cl (100mL). The resulting solution is extracted with 3x100mL ethyl acetate and the combined organic layer is dried over Na2SO4 and concentrated under vacuum. The crude solid is washed with ethyl acetate / hexane (1:5) (2x 100mL). This resulted in 6.81 g (90%) of the title compound as a yellow solid. MS-ESI: 337.1 (M+1), 335.1 (M-1), in positive and negative ion modes, respectively.

[0208] Step 6: N'-(tert-butyldimethylsilyl)-5-(2-hydroxypropan-2-yl)thiazole-2-sulfonylimideamide

[0209] PPh is put into the 100-mL 3-neck round-bottom flask purged with nitrogen and maintained under nitrogen Cl (3g, 9.0mmol) solution in CHCl (100mL). DIEA (1.54g, 11.9mmol) is added dropwise with stirring at RT subsequently. The resulting solution is stirred for 10min at RT. N-(tert-butyldimethylsilyl)-5-(2-hydroxypropyl-2-yl)thiazole-2-sulfonamide (2.0g, 5.9mmol) is added dropwise with stirring in an ice / water bath subsequently (30mL). The resulting solution is stirred for 30min in an ice / water bath. NH is introduced into the above solution at a temperature lower than 0°C (g), and continues for 15 minutes. The resulting solution is stirred for 20 minutes at RT. Solid is filtered out, and the filtrate is concentrated and the residue is dissolved in 300mL ethyl acetate. The solution was washed with brine (2x 100mL), dried over Na2SO4 and concentrated under vacuum. The crude solid was washed with CHCl3 (100mL). The filtrate was then concentrated under vacuum and the residue was further purified by silica gel column with ethyl acetate / petroleum ether (1:10 to 1:3). The solid initially washed was merged with the solid from silica gel purification. This produced 1.2g (60%) of the title compound as a white solid. MS-ESI: 336.1 (M+1). 1H-NMR (300MHz, DMSO-d6) δ7.66(s,1H),7.12(s,2H),5.78(s,1H),1.51(s,6H),0.86(s,9H),0.02(s,3H),0.01(s,3H).

[0210] The following abbreviations have the indicated meanings:

[0211] ACN = acetonitrile

[0212] BTC = trichloromethyl chloroformate

[0213] Boc=tert-butyloxycarbonyl

[0214] Davephos = 2-dicyclohexylphosphino-2'-(N,N-dimethylamino)biphenyl

[0215] DCM = dichloromethane

[0216] DEA = Diethylamine

[0217] DMF=N,N-dimethylformamide

[0218] DMSO = dimethyl sulfoxide

[0219] DIEA=N,N-diisopropylethylamine

[0220] DPPA = diphenylphosphoryl azide

[0221] dppf=1,1'-bis(diphenylphosphino)ferrocene

[0222] EtOH = ethanol

[0223] HATU = 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate

[0224] Hex = Hexane

[0225] HPLC = High Performance Liquid Chromatography

[0226] LC-MS = Liquid Chromatography-Mass Spectrometry

[0227] LiHMDS = Lithium Bis(trimethylsilyl)amide

[0228] LDA = Lithium diisopropylamide

[0229] M=mol / L

[0230] Me = methyl

[0231] MeOH = methanol

[0232] MSA = Methanesulfonic acid

[0233] NBS = N-bromosuccinimide

[0234] NCS = N-chlorosuccinimide

[0235] NMR = Nuclear Magnetic Resonance

[0236] Pd(dppf)Cl2=Dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium

[0237] Ph = Phenyl

[0238] PPh3Cl2=triphenylphosphine dichloride

[0239] Py = Pyridine

[0240] RT = Room temperature

[0241] Rt = retention time

[0242] R f = Retardation coefficient

[0243] Sat.=Saturated

[0244] TBAF = Tetrabutylammonium fluoride

[0245] TBS = tert-butyldimethylsilyl

[0246] TBSCl = tert-butyldimethylsilyl chloride

[0247] TBDPSCl = tert-butyldiphenylsilyl chloride

[0248] TEA = triethylamine

[0249] TFA = trifluoroacetic acid

[0250] THF = Tetrahydrofuran

[0251] TLC = Thin Layer Chromatography

[0252] TsOH = 4-methylbenzenesulfonic acid

[0253] UV = Ultraviolet light

[0254] bid = twice a day

[0255] WCC = white blood cell count

[0256] EP = End Point

[0257] y = years old

[0258] y / n = yes / no

[0259] Examples

[0260] The following examples illustrate the methods and uses described herein. However, they are not intended to limit the scope of the methods and uses described in any way. Other variations of the embodiments will be clear to those of ordinary skill in the art and are covered by the appended claims.

[0261] Example 1: Clinical study with compound IA

[0262] An open-label, single-arm Phase II study of Compound IA to evaluate the safety, tolerability, and efficacy in participants with familial cold autoinflammatory syndrome (FCAS) who have evidence of inflammatory activity following a cold challenge during the screening period. Figure 1 is an overview of the treatment regimen with Compound IA (ie, the R enantiomer of Compound 1). Compound IA 100 mg will be administered twice daily for 3 days, and 100 mg will be administered in the morning of the 4th day.

[0263] The primary objective of this study was to evaluate the efficacy of Compound IA in reducing cold-induced inflammation in participants with FCAS.

[0264] The primary objective endpoint (EP) was the change in white blood cell count (WCC) from pre-challenge to the highest post-challenge value.

[0265] The secondary objectives of the study are as follows:

[0266] 1. Evaluation of the safety and tolerability of compound IA

[0267] 2. Evaluate the efficacy of compound IA in improving the signs and symptoms of FCAS

[0268] 3. Evaluation of the effect of compound IA on patient-reported outcomes

[0269] The secondary endpoints (EP) are as follows:

[0270] 1. Safety endpoints (including vital signs, ECG parameters, safety laboratory evaluations, and adverse events)

[0271] 2. Changes from pre-challenge to post-challenge in physicians' gross assessment of autoinflammatory disease activity and physicians' assessment of severity of autoinflammatory disease signs and symptoms

[0272] 3. Changes in patients’ gross assessment of disease activity from pre-challenge to post-challenge

[0273]

[0274] Study Design

[0275] This is an open-label, single-arm, multiple-dose study in approximately six participants with FCAS and confirmed NLRP3 gain-of-function mutations who show evidence of inflammatory activity following a cold challenge performed during the screening period.

[0276] The cold challenge protocol was developed to investigate acute inflammatory mechanisms in patients with FCAS following general cold exposure and to investigate the effect of pretreatment with IL-1 blockade therapy. Participants with a history of transient self-limited rash, fever, and / or arthralgia who exhibit characteristic symptoms of FCAS following cold exposure and who resolve after warming will undergo up to two controlled cold challenges to induce a transient inflammatory response while residing at the clinical site under medical supervision. Following the 45-minute cold challenge, participants will return to an ambient temperature of approximately 25°C at the clinical site for at least 24 hours to ensure resolution of the inflammatory response.

[0277] Typical symptoms and signs induced by cold challenge in participants with FCAS include increases in systemic inflammatory markers (e.g., WCC, neutrophils), low-grade fever, rash, and arthralgia, which begin 1 hour after challenge, peak at 4-8 hours, and abate by the next day, with no reports of long-term sequelae (Hoffman et al., Prevention of cold-associated acute inflammation in familial cold autoinflammatory syndrome by interleukin-1 receptor antagonist. Lancet; 2004; 364(9447): 1779-85.).

[0278] The study covered three periods:

[0279] Screening period, including screening cold challenge (up to three months)

[0280] Treatment period, including second cold challenge (duration 5 days total)

[0281] Follow-up period, ending with the study completion visit (10 days after dosing)

[0282] Screening period, to confirm compliance with study inclusion and exclusion criteria, and to conduct clinical observations and biological sampling:

[0283] Screening Visit: Informed consent will be collected and participant eligibility will be assessed. For those participants who qualify for the Screening Cold Challenge, prohibited medications will be discontinued. Results of laboratory specimens collected at the Screening Visit and the participant's active disease report must be obtained prior to scheduling the Screening Cold Challenge.

[0284] Screening Cold Challenge: Participants will reside on site for three days associated with the Screening Cold Challenge. On the first day (Day -10), participants will be admitted in the morning, undergo a safety assessment, and then stay in a room at ambient temperature (preferably above 25°C) to ensure stable conditions prior to the cold challenge. On the second day (Day -9), after the pre-challenge assessment and breakfast, participants will undergo a 45-minute cold challenge and then be monitored for the next 23 hours. On the third day (Day -8), participants will be discharged after the last assessment, or later at the discretion of the investigator. Participants should have at least one week to recover from the cold challenge prior to recruitment and initiation of treatment.

[0285] Treatment period

[0286] Treatment Initiation Visit: Participants who meet all inclusion criteria and no exclusion criteria (including showing evidence of inflammatory activity following a cold challenge during the screening period) will be enrolled on Day 1 and begin treatment.

[0287] The first dose of Compound IA will be administered in the clinic, and participants will be assigned study treatment to continue at home. Participants may reside in the clinic during treatment at the discretion of the participant and the investigator for reasons of convenience to the participant and / or organizational arrangements.

[0288] Cold Challenge: Participants will be admitted to the clinic on the morning of the 3rd day (i.e. the day before the cold challenge), and if required by the investigator or local regulations, participants will live for a total of three days or more. On the 3rd day, participants will receive a safety assessment and then stay in a room at ambient temperature (preferably above 25°C) to ensure that conditions are stable before the cold challenge. On the morning of the 4th day, a pre-challenge assessment will be performed and breakfast will be provided. Pre-dose PK samples will be collected immediately after breakfast, and the last dose of study treatment will be administered. One hour after the study treatment is administered, another PK sample will be collected and a cold challenge will be started. The cold challenge lasts 45 minutes. After the cold challenge ends, participants will be monitored until the next morning (Day 5). At the discretion of the investigator, participants will be discharged after the last assessment on or after Day 5.

[0289] A follow-up period was used to ensure the safety of participants after they stopped treatment and recovered from the cold challenge.

[0290] Participants will be followed up for end-of-study evaluations at the end-of-study visit, approximately 10 days after the last dose.

[0291] Key inclusion criteria

[0292] 1. Written informed consent must be obtained before any study-specific assessments are performed

[0293] 2. Male and female participants aged between 18 and 80 years old (inclusive)

[0294] 3. Body mass index is between 18-35 kg / m 2 within the scope

[0295] 4. Participants genetically diagnosed with FCAS

[0296] 5. Participants with a clinical history and findings consistent with FCAS in the absence of a history or diagnosis of amyloidosis and / or organ damage (e.g., deafness, periorbital edema, lymphadenopathy, and serositis)

[0297] 6. Participants with evidence of inflammatory activity after screening cold challenge

[0298] 7. Autoinflammatory disease activity > minimal based on gross physician assessment, with clinical history of active disease in response to cold exposure in the environment in at least one setting during the screening period

[0299] Key exclusion criteria

[0300] 1. Participants with NLRP3 mutations that are unresponsive to NLRP3 inhibition, including but not limited to L353P carriers (based on principal investigator / registry data, published evidence, and / or internal studies at Novartis AG)

[0301] 2. Participants currently receiving anti-rejection and / or immunomodulatory medications must not interrupt treatment or switch to other agents before the screening cold challenge and for the duration of the study within 28 days or 5 half-lives (whichever is longer if required by local regulations) or before the expected pharmacodynamic effect returns to baseline for the immunomodulatory therapeutic antibody

[0302] Exceptions are: anakinra, canakinumab, and / or other investigational IL-1 / NLRP3 binding or blocking therapies, which must be discontinued at screening. Once evidence of active disease is met, participants can proceed to the screening cold challenge.

[0303] 3. Clinically significant, suspected active or chronic bacterial (including Mycobacterium tuberculosis), viral or fungal infection within 30 days prior to administration

[0304] 4. Participants with congenital (e.g., TLR immunodeficiency, IFN-γ signaling defect) or acquired immunodeficiency (e.g., AIDS)

[0305] 5. Presence of human immunodeficiency virus (HIV) infection, hepatitis B surface antigen (HBsAg) or hepatitis B core antibody (anti-HBc) or hepatitis C antibody at screening

[0306] 6. Live vaccine within 4 weeks of Day 1 (i.e., first dose of Compound IA)

[0307] 7. Absolute peripheral blood neutrophil count ≤ 1000 / mm 3

[0308] 8. Estimated glomerular filtration rate (eGFR) ≤ 90 mL / min / 1.73 m2 (based on the Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) formula)

[0309] 9. History of ECG abnormalities or current diagnosis indicating a significant safety risk for enrolled participants

[0310] 10. Pregnant or lactating (lactating) females, where pregnancy is defined as the state of a female from conception until termination of pregnancy, as determined by a positive human serum chorionic gonadotropin (hCG) laboratory test

[0311] 11. Females of childbearing potential, defined as all females who are physiologically capable of becoming pregnant, unless these females use highly effective contraceptive methods during dosing and within 10 days after stopping study treatment

[0312] 12. Any significant concurrent medical condition that the investigator believes may affect the participant's ability to tolerate or complete the study

[0313] Efficacy evaluation:

[0314] White blood cell count (WCC)

[0315] Physician's gross assessment of autoinflammatory disease activity

[0316] Physician severity assessment of signs and symptoms of autoinflammatory disease

[0317] · Patient's general assessment of disease activity

[0318] Key safety assessments

[0319] Physical examination

[0320] Vital signs

[0321] ECG parameters

[0322] Pregnancy and fertility assessment

[0323] Monitoring of laboratory markers in blood and urine

[0324] Adverse event and serious adverse event monitoring

[0325] Other assessments

[0326] Inflammatory markers (including but not limited to absolute neutrophil count (ANC), high-sensitivity C-reactive protein (hsCRP), and serum amyloid A (SAA))

[0327] Plasma concentration and pharmacokinetic parameters of compound IA

[0328] Pharmacodynamics, inflammasome pathways, and inflammation-related biomarkers in serum, such as IL-1β, IL-6, IL-18, IL-18 binding protein (IL-18BP), CXCL10, caspase-1, and proteomics

[0329] Data analysis

[0330] The primary objective will be achieved and Compound IA will be considered effective in treating cold-induced inflammation in participants with FCAS if the estimated difference in the change in total WCC from pre-challenge between the treatment and screening periods meets the following criteria:

[0331] 1. Statistically significant (p<0.10) and

[0332] 2. Less than -4 x 109 cells / L

[0333] Changes in maximum WCC from pre- to post-challenge will be analyzed by linear models including pre-challenge values ​​as covariates and modeling period effects, taking into account within-participant correlations.

[0334] Rationale for dose / regimen, duration of treatment

[0335] Based on preliminary PK results and preliminary PK / PD relationships, a dose of 100 mg administered twice daily as a tablet was selected for this study.Compound IA tablets will be administered shortly after completion of a meal, and all doses will be administered approximately 12 h (+ / - 1 h) apart.

[0336] The 100 mg tablet had a positive food effect on PK (peak plasma levels (Cmax) increased 2.05-fold and AUClast increased 1.49-fold). The apparent terminal elimination half-life of Compound IA tablets under fed conditions was approximately 10 hours, so steady state is expected to be reached on day 4 after multiple dosing.

[0337] There is no established occupancy marker for NLRP3. Ex vivo whole blood, LPS-stimulated IL-1β secretion data from FIH studies have been used to estimate effective doses. Based on the results of this assay, the average total Compound IA plasma trough concentration (IC90) required to inhibit 90% of stimulated IL-1β release in healthy participants averaged 3.17 μM, equivalent to approximately 1.3 μg / mL.

[0338] Based on the 3-5-fold greater in vitro IC50 values ​​for inhibition of IL-1β secretion measured in PBMCs of patients with certain FCAS-associated mutations compared to healthy participants, higher concentrations may be required in participants with FCAS. This may be due to mutations in the NLRP3 protein affecting the binding KD of the compound. Assuming that plasma levels 5-fold higher than the IC90 values ​​derived from preliminary PK / PD studies are required over a 24-hour period to maintain full target occupancy in participants with FCAS, a dose of 100 mg twice daily administered shortly after completing a meal would be used to ensure complete inhibition of NLRP3 by day 4.

[0339] In a human study of healthy volunteers, a = once daily administration of 200 mg doses of Compound 1A for up to 14 days, the mean AUC0-24h and Cmax at steady state were 182 μg*h / mL and 26.4 μg / mL, respectively. For twice daily administration of 100 mg tablets, the predicted steady-state mean AUC0-24h (225 μg*h / mL) was 1.2 times higher than the corresponding PK parameters of healthy volunteers administered 200 mg Compound 1A, and Cmax (12.1 μg / mL) was 2.2 times lower.

[0340] A 13-week Good Laboratory Practice (GLP) toxicology study in rats and cynomolgus monkeys supported the safety of the treatment period.

[0341] Table 1

[0342]

[0343] a Based on total exposure, for unbound exposure (corrected for plasma protein binding), the safety margin was higher in rats (2-fold) and lower in monkeys (0.8-fold).

[0344] b The NOAEL Cmax (μg / mL) and AUC0-24h (μg*h / mL) of a 13-week toxicology study in rats (30 mg / kg) were 52.9 and 400, and for monkeys (150 mg / kg) were 258 and 3540, respectively.

[0345] Considering the short treatment duration of 4 days and the adequate safety margin, the administration of Compound IA in this study was considered safe.

[0346] Drug-drug interaction issues:

[0347] Evaluation and recommendations for clinical drug-drug interaction studies of cytochrome P450 (CYP) substrates / modulators and Compound IA are based on in vitro / preclinical data and physiologically based PK simulations. Compound IA is expected to be eliminated primarily via hepatic CYP-mediated metabolism, with CYP2C9 (68%) and CYP3A4 (29%) being the major contributing enzymes. Participants who are poor CYP2C9 metabolizers will be excluded from this study.

[0348] Considering the duration of treatment and the adequate safety margin, administration of Compound IA was considered safe even under conditions of elevated Compound IA exposure.

[0349] Prohibited drugs and herbal remedies

[0350] Anti-rejection / immunomodulatory therapy (eg, anakinra, canakinumab, or other investigational IL-1 / NLRP3 binding or blocking therapies)

[0351] Live vaccines

[0352] Strong or moderate inducers of CYP2C9 or strong inducers of CYP3A, including carbamazepine, enzalutamide, lumacato, phenobarbital, phenytoin, rifabutin, mitotane, and St. John's wort (Hypericumperforatum)

[0353] Strong inhibitors of CYP2C9, including miconazole, berberine (herbal product), sulfaphenazole, fluconazole, and resveratrol (herbal product)

[0354] Other research products

[0355] Use drugs with caution

[0356] Drugs metabolized by CYP3A

[0357] In vitro metabolism studies have shown that Compound IA may have the potential to induce the metabolism of drug substrates metabolized by the isozyme CYP3A. Therefore, concomitant agents known to be metabolized by CYP3A4 / 5 may be administered at the investigator's discretion. Patients receiving such agents may require dose adjustments or increases in concomitant medications. In particular, caution is advised when Compound IA is co-administered with drugs that are sensitive substrates of CYP3A and / or have a narrow therapeutic index.

[0358] Drugs that are strong or moderate inhibitors of CYP3A

[0359] Compound IA was identified as a CYP3A substrate in vitro, therefore an increase in systemic exposure of Compound IA cannot be excluded when co-administered with strong CYP3A inhibitors such as antivirals (e.g., ritonavir), antifungals (e.g., itraconazole, ketoconazole), and antibiotics (e.g., erythromycin, clarithromycin). Co-administration of known inhibitors of CYP3A may be done at the investigator's discretion, but its duration should be kept as short as possible and the patient must be closely monitored.

[0360] Example 2: First-in-human (FIH) clinical study of compound IA:

[0361] Study Design

[0362] The study design consisted of 4 parts: single ascending dose (SAD; Part A), relative bioavailability of the tablet formulation (Part B), multiple ascending dose (MAD; Part C), and relative bioavailability and food effect (Part D) ( Figure 2 In each group of Parts A and C, 8 subjects were randomized in a 3:1 ratio to receive Compound IA (6 subjects) or matching placebo (2 subjects).

[0363] For Part A, a total of 8 groups with 8 eligible subjects were recruited. Each subject received a single oral dose of Compound IA (3, 10, 30, 100, 300 mg crystalline suspension and 100, 300, 600 mg SDD under fasting conditions). In Group 1 of Part A, 2 sentinel subjects were dosed at least 24 hours prior to dosing of the remaining subjects in the group to ensure maximum safety.

[0364] Part B was skipped as the data collected in Part A provided an adequate comparison of the crystalline formulation with the SDD formulation.

[0365] In Part C, eligible subjects were recruited into 6 different groups. Each subject received multiple doses of Compound IA (10, 30 mg crystalline suspension and 100, 200 mg SDD suspension) once a day (QD) in the fasting state for 14 days, or received multiple doses of Compound IA (25, 50 mg encapsulated crystalline tablets for 13 days, and received a single dose or placebo on Day 14) twice a day (BID) under fed conditions. Subjects in Part C were dosed after reviewing the available safety, tolerability and PK data from the previous groups in Part A.

[0366] Part D had an open-label, randomized, 3-period crossover design consisting of 1 group with 6 subjects. The PK profile of the crystalline tablet formulation evaluated in the nonclinical safety pharmacology and toxicology program of Compound IA was compared between fed and fasted conditions and compared to the PK profile of the crystalline suspension of Compound IA under fasted conditions. Subjects received 3 doses of Compound IA with a washout period of 7-14 days between each dose (Dose 1: 100 mg oral suspension under fasted conditions; Dose 2: 100 mg oral tablet under fasted conditions; Dose 3: 100 mg oral tablet under fed conditions). Based on these doses, subjects were randomly assigned to 1 of 6 treatment sequences (1 subject per sequence) prepared using a Williams design (Wang BS, Wang XJ, Gong LK. The Construction of a Williams Design and Randomization in Cross-Over Clinical Trials Using SAS. Journal of Statistical Software 2009; 29).

[0367] Subjects

[0368] Eligible subjects were between 18 and 64 years of age with a body mass index (BMI) ≥ 18.5 and ≤ 30.0 kg / m 2 Healthy volunteers. No subject participated in >1 part or group. Written informed consent was obtained before any study procedures. Subjects participating in Part D had to be willing and able to consume a full high-fat breakfast within the specified time frame. Subjects were excluded if they had a history of severe psychiatric disorder, a diagnosis of intellectual disability, clinically significant abnormal vital signs, or used tobacco products within 90 days before (first) drug administration until follow-up.

[0369] Blind

[0370] In Part A and Part C, active and placebo treatments could not be distinguished based on labeling, they looked identical, and had similar taste and smell. To maintain the blind, each subject in the corresponding group was given the same number of tablets or suspensions. Investigators and subjects remained blind throughout the relevant parts of the study, and the blind was maintained without breaking throughout the process. The sponsor remained unblinded for all study data and obtained a copy of the randomization code to support decisions about the study. Part D was open-label according to the Williams design, with compound IA administered only to subjects in 1 of 6 treatment sequences (1 subject per sequence).

[0371] Target

[0372] The primary objective of the study was to evaluate the safety and tolerability of SAD and MAD oral doses of Compound IA in healthy subjects in all parts of the study. Key secondary objectives were to characterize the PK profile of Compound IA after single and multiple doses and to evaluate the effect of food on the PK profile of Compound IA.

[0373] Evaluate

[0374] Safety Assessment

[0375] Safety assessments in all parts of the study included adverse events (AEs) reported using the Medical Dictionary for Regulatory Activities (version 22.1), clinical laboratory tests (biochemistry, hematology, and urinalysis), vital signs, electrocardiogram (ECG), physical examination, and skin biopsy (if applicable).

[0376] Pharmacokinetic (PK) evaluation

[0377] In the single-dose portion, blood samples were collected at the following time points relative to dosing on Day 1 to determine the concentration of Compound IA: before dosing and 0.25, 0.5, 0.75, 1, 1.5, 2, 3, 4, 6, 8, 12, 24, 36 and 48 hours after dosing, and at follow-up. In the multiple-dose portion, blood samples were collected at the following time points relative to dosing on Days 1 and 14: before dosing and 0.25, 0.5, 0.75, 1, 1.5, 2, 3, 4, 6, 8 and 12 hours after dosing; on Days 2, 4, 7, 9 and 11: before dosing; after the last dose on Day 14: 24 and 36 hours (Day 15) and 48 hours (Day 16) after dosing; and at follow-up.

[0378] In the multiple ascending dose (MAD) portion, urine pools were collected over 24 hours for QD doses (0-12h and 12-24h) and over 12 hours for BID doses relative to dosing on days 1 and 14. Plasma samples were analyzed by fully validated liquid chromatography-tandem mass spectrometry (LC-MS / MS). The assay precision for compound IA concentrations >1 ng / mL (lower limit of quantification, LLOQ) was ≤4.4%, and the relative error of accuracy was -2.0% to 1.5%. Urine samples were analyzed by LC-MS / MS with an LLOQ of 1 ng / mL. Concentrations below the LLOQ were set to 0.

[0379] The following PK parameters were estimated using non-compartmental analysis: maximum concentration in plasma (C max ); time to reach maximum concentration (t max ); concentration 24h after administration (C 24h )(Part A); Lag time: the time before the first quantifiable concentration is observed (t lag ); time to reach the last quantifiable concentration (t last ); the area under the concentration-time curve (AUC) from time 0 to the last quantifiable concentration 0-last ); the area under the plasma concentration-time curve from time 0 to infinity (AUC 0-inf ); the area under the plasma concentration-time curve from time 0 to 24 hours after administration (AUC 0-24 ); terminal phase rate constant (K el ); terminal half-life (t 1 / 2 ); apparent total clearance (CL / F); and apparent volume of distribution at the terminal stage (V z / F); in addition, for Part C only: the area under the plasma concentration-time curve (AUC 0-tau ); Apparent clearance at steady state (CLss / F); Based on AUC 0-tau The cumulative ratio (R ac , AUC); and based on C max The cumulative ratio (R ac ,C max ).

[0380] Pharmacodynamic (PD) evaluation

[0381] To determine the PD response to the NLRP3 inhibitory pathway, whole blood samples were collected for exploratory PD analysis (groups 1-3 in parts A and C). The inflammatory marker IL-1β was analyzed after ex vivo stimulation of whole blood samples with lipopolysaccharide (LPS). Whole blood stimulation with 1 μg / mL LPS was successful, and analysis of IL-1β concentrations in plasma samples using a validated electrochemiluminescence assay was found to be valid and scientifically acceptable. The LLOQ after dilution was 64.6 ng / L.

[0382] Statistical analysis

[0383] All data were summarized using descriptive statistics and presented and summarized in tables and / or graphs. Descriptive statistics for all relevant PK parameters included: n, arithmetic mean, standard deviation (SD), coefficient of variation (CV%), minimum, median, maximum, geometric mean, and geometric CV%. For t max Only median, minimum and maximum values ​​are presented. PK parameters were calculated using non-compartmental methods using Phoenix version 8.1 software. Concentrations below the lower limit of quantitation were treated as zero in the summary statistics of the individual concentration data. The linear trapezoidal rule was used for AUC calculations. For determination of t 1 / 2 Regression analysis of the terminal plasma elimination period included C max At least 3 data points after the adjustment. 2 Parameters below 0.80 are flagged, but are included in the descriptive statistics. Parameter AUC 0-inf , %AUC extra , CL / F and %AUC extra VZ / F above 20% were excluded from descriptive statistics.

[0384] In part A, the log-transformed C max , AUC 0-last and AUC 0-inf Dose proportionality was explored using a regression power model associated with log-transformed dose levels. Point estimates of the intercept and slope and the corresponding 90% confidence interval (CI) for the slope were calculated. For Part C, dose proportionality was not explored. In Part D, an analysis of variance (ANOVA) model for the PK data was used to explore the relative bioavailability of the crystalline tablet relative to the crystalline suspension and the food effect. The ANOVA model included fixed effects for treatment, period, and sequence, and a random effect for subjects within the sequence.

[0385] Least squares geometric mean ratios are presented together with 90% CI for the following treatments: 100 mg Compound IA tablet when fasted versus 100 mg Compound IA suspension when fasted, and 100 mg Compound IA tablet when fed versus 100 mg Compound IA tablet when fasted.

[0386] Combined individual and mean graphs of IL-1β concentrations versus time are presented by treatment.

[0387] To characterize the inhibitory potency of compound IA on IL-1β release, a Bayesian Emax PK / PD model was applied. The model was developed in the free software environment R (version 4.0.5) using the rstan library. Convergence issues (probably related to the fitting of baseline values ​​and inter-individual variability) were the final logical justification for choosing a Bayesian approach over a frequentist approach, which was also initially evaluated—and the reason for including only one parameter with inter-individual variability. Two results are presented: one estimating the maximum inhibition and the other assuming the ability to achieve complete inhibition (E max =1). Ex vivo LPS-stimulated results were corrected for unstimulated results of the same sample. Concentrations below the LLOQ were set to half the LLOQ (IL-1β) and 0 (Compound IA).

[0388] result

[0389] Subject disposition and demographics

[0390] A total of 122 subjects were enrolled in the study. All 122 subjects were included in the safety and PD analysis sets, and 94 subjects who received active treatment (Compound IA) were included in the PK analysis set. Overall, subjects aged between 18 and 64 years and with a BMI between 18.9 and 29.4 kg / m 2 Fifty-eight (48%) males and 64 (52%) females participated in the study. The majority (105 [86%]) of the subjects (Part A, n=57; Part C, n=42; Part D, n=6) were Caucasian.

[0391] Of the subjects recruited, 107 (88%) subjects completed the study as per protocol, and 15 (12%) subjects discontinued the study early. Early discontinuation included 1 (2%) of 64 subjects in Part A, 13 (25%) of 52 subjects in Part C, and 1 (17%) of 6 subjects in Part D. The reasons for discontinuation of the study included 12 (10%) subjects withdrawing due to AEs, of which 1 (1%) subject each discontinued the study due to withdrawal of consent, loss of follow-up, or suspension of the study due to the COVID-19 pandemic (preventing visits; unrelated to the safety of compound IA). A total of 4 discontinued subjects were replaced in Part C.

[0392] Security

[0393] The tolerability of single and multiple doses of Compound IA was generally good. No deaths or serious AEs (SAEs) were reported during the study. Overall, 87 of 122 subjects (71%) reported treatment-emergent AEs (TEAEs): 66 of 94 subjects in the Compound IA group (70%) and 21 of 28 subjects in the placebo group (75%) reported treatment-emergent AEs (TEAEs). Most TEAEs reported by 84 (69%) subjects were of mild intensity, while 15 subjects (12%) reported moderate TEAEs. System organ class (SOC) events frequently reported in >20% of subjects were nervous system disorders (34%), general disorders and administration site conditions (29%), and gastrointestinal disorders (27%).

[0394] Overall, 46 relevant TEAEs reported by 24 of 122 subjects (20%) were considered related to study drug, including 21 of 94 subjects (22%) receiving Compound IA and 3 of 28 subjects (11%) receiving placebo. For 12 of 122 subjects (10%), 20 TEAEs of maculopapular rash and / or pruritus were considered AEs of particular concern because they were considered related to study drug. All 12 subjects received Compound IA, either as a single dose (100 mg CS [n=1] or 600 mg SDD [n=1]) or in multiple doses (30 mg QD CS [n=2], 100 mg QD SDD [n=3], 200 mg QD SDD [n=2], or 50 mg BID ECT [n=3]). These TEAEs were mild to moderate in intensity, generally began within 1-17 days after initiation of treatment with Compound IA, and resolved within 1-18 days after onset; all cases resolved without the need for concomitant treatment. For 10 subjects, these TEAEs led to treatment discontinuation. Two additional subjects discontinued early due to TEAEs unrelated to study drug.

[0395] Mild decreases in neutrophil and leukocyte counts were considered nonclinically significant and were only occasionally noted, which may be consistent with the PD effects of Compound IA caused by inhibition of IL-1β signaling downstream of NLRP3. One subject was considered to have second-degree atrioventricular block unrelated to study drug. No other clinically relevant findings were reported on vital signs, 12-lead ECG, 24-hour Holter monitoring, or physical examination.

[0396] Pharmacokinetics

[0397] As shown by dose-normalized Cmax and AUC, when Compound IA was administered as a crystalline suspension, plasma exposure of a single dose of Compound IA increased in a less-than-dose-proportional manner (3-300 mg, slope [90% CI] for Cmax: 0.518 [0.460; 0.577], and for AUC0 last, 0.701 [0.614; 0.788]). Although at lower dose levels (3 mg to 30 mg), mean AUC 0-last appears to increase in proportion to dose, but at the 100 mg and 300 mg dose levels, the mean AUC 0-last The exposures were similar (79500 (CV 54.8%) and 64400 ng·h / mL (CV 23.5%), respectively). When administered as an SDD suspension, exposure increased proportionally with dose (100-600 mg, slope [90% CI]: 0.913 [0.802; 0.1.024] for Cmax and 0-last For example, 1.121[0.977;0.1.265]).

[0398] Following QD administration of Compound IA at doses ranging from 30 to 200 mg for 2 weeks, only limited drug accumulation of approximately 1.1- to 1.3-fold was observed upon reaching steady state. This is consistent with the average t ranging from 9.83 to 16.2 hours at both QD and BID dose levels. 1 / 2 At steady state, Compound IA exhibited very low CLss / F (approximately 0.83 to 1.11 L / h) and Vss / F (approximately 12.6 to 23.3 L), with low to moderate inter-subject variability at both QD and BID dose levels of Compound IA.

[0399] The total cumulative amount of compound IA excreted in urine increased linearly with increasing multiple dose levels. Compound IA was mostly excreted within 12 hours. On days 1 and 14, the mean dose fraction excreted was in the range of 0.3%-0.4% and 0.7%-1.1%, respectively, and the renal clearance was in the range of 3.6-5.0 mL / h and 6.8-9.1 mL / h, respectively.

[0400] Administration of a single dose of 100 mg of Compound IA as a crystalline suspension under fed conditions resulted in a decrease in the C of Compound IA compared to fasting conditions. max Increased by 2.05 times and AUC 0-last For the crystalline tablets (100 mg Compound IA under fasting conditions), the median t max Delay from 2 hours to 5 hours, C max The AUC was similar (104%) and there was a significant difference between the crystalline tablet and the suspension. 1 / 2 The encapsulated crystalline tablets (25 and 50 mg BID under fed conditions) were characterized by median lag times of 0.75 and 0.25 hours, respectively, and median t max The average t of compound IA is 4 hours. 1 / 2 It was comparable between tablet (18.6 hours) and suspension (17.7 hours) formulations.

[0401] Pharmacodynamics

[0402] Dose-dependent decreases in ex vivo stimulated IL-1β concentrations were observed with increasing single and multiple oral doses of Compound IA (mean nadir concentrations were approximately 5%-20% of baseline values). At most dose levels of Compound IA, inhibition of IL-1β release was observed from 1 hour post-dose until the last sampling time point - single (Day 3 or up to 6 hours for the lowest ≤10 mg dose level) and multiple (Day 15) oral doses of Compound IA.

[0403] Based on the fractional maximum stimulation (E max ) model, the typical baseline (E0) (±SD) of IL-1β stimulation observed was 1820 (±102) ng / L; the E max The effective concentration for the estimated maximal effect of compound IA caused by ex vivo stimulated IL-1β release is EC 50 :59ng / mL (90% CI: 48,72), EC 90:1080ng / mL (90% CI: 942,1240). When considering complete inhibition (fixed E max =1), the plasma concentration of compound IA that inhibited 50% and 90% of the IL-1β release stimulated by LPS in vitro (IC 50 and IC 90 ) were 61 ng / mL (90% CI: 50; 70) and 1340 ng / mL (90% CI: 1190; 1490), respectively, and the Hill coefficient was 0.715 (± 0.0333). The similarity between the models suggests that replacing values ​​below LOQ with LOQ / 2 has limited impact.

[0404] discuss

[0405] There is an unmet medical need to expand the therapeutic modality of providing an effective and predictable treatment option without an increased risk of AEs for patients suffering from inflammasome NLRP3-mediated inflammatory, metabolic, and neurodegenerative diseases. In this study, the NLRP3 antagonist compound IA was orally administered to human subjects for the first time to explore its safety, tolerability, PK, and PD properties. The initial dose was selected based on the predicted human PK and expected effective dose, as well as nonclinical safety from animal and in vitro data.

[0406] Security

[0407] Single and multiple doses of Compound IA or placebo were generally well tolerated. No deaths or SAEs were reported during the study. Similar TEAE rates were observed between subjects receiving Compound IA (70%) and placebo (75%). Most TEAEs reported by subjects were mild (69%) or moderate (12%) in severity. Subcutaneous tissue and gastrointestinal disorders TEAEs were reported only by subjects receiving Compound IA and not by subjects receiving placebo. Maculopapular and / or pruritic rash was most frequently reported at higher multiple dose levels of Compound IA, indicating a relationship with exposure to Compound IA and not with the formulation used. Safety and tolerability data of other NLRP3 inhibitors such as ZYIL1 or dapasuntrile tested in clinical trials (Parmar DV, Kansagra KA, Momin T, Patel HB, Jansari GA, Bhavsar J, Shah C, Patel JM, Ghoghari A, Barot A, Sharma B, Viswanathan K, Patel HV, Jain MR. Safety, Tolerability, Pharmacokinetics, and Pharmacodynamics of the Oral NLRP3 Inflammasome Inhibitor ZYIL1: First-in-Human Phase 1 Studies (Single Ascending Dose and Multiple Ascending Dose). Clin Pharmacol Drug Dev. Clinical Pharmacology in Drug Development 2023;12:202-211;Marchetti C,Swartzwelter B,Gamboni F,Neff CP,Richter K,Azam T,Carta S,Tengesdal I,Nemkov T,D'Alessandro A,Henry C,Jones GS,Goodrich SA,Laurent JP,Jones TM,Scribner CL,Barrow RB,Altman RD,Skouras DB,Gattorno M,Grau V,Janciauskiene S,Rubartelli A,Joosten LAB,Dinarello CA.OLT1177, a β-sulfonylnitrile compound, safe in humans, inhibits the NLRP3 inflammasome and reverses the metabolic cost of inflammation. Proc Natl Acad Sci USA. 2018;115:E1530-E1539; Klück V, Jansen TLTA, Janssen M, Comarniceanu A, Efdé M, Tengesdal IW, Schraa K, Cleophas MCP, Scribner CL, Skouras DB, Marchetti C, Dinarello CA, Joosten LAB. Dapansutrile, an oral selective NLRP3 inflammasome inhibitor, for treatment of gout flares: an The open-label, dose-adaptive, proof-of-concept, phase 2a trial (Dapansutrile is an oral selective NLRP3 inflammasome inhibitor for the treatment of gout flares: an open-label, dose-adaptive, proof-of-concept, phase 2a trial) did not include any drug-related skin reactions, pointing to specific Compound IA-related effects rather than due to mode of action.

[0408] Pharmacokinetics

[0409] After a single oral dose of Compound IA in suspension form (3-300 mg CS, 100-600 mg SDD) under fasting conditions, Compound IA was generally rapidly absorbed, with a median t max However, at the higher dose range of 30–600 mg, the median t maxThere was a slight delay (1.5-3.0 hours). The increase in drug exposure was less proportional to the dose for the crystalline suspension (particularly 100 and 300 mg), whereas a dose-proportional increase in exposure was observed with the SDD suspension (100-600 mg), indicating that absorption of the crystalline material was solubility limited at doses ≥ 100 mg.

[0410] Multiple doses and formulations of Compound IA showed no deviation from dose proportional drug exposure after 2 weeks, indicating that multiple dose PK parameters were linear and not solubility limited. Following an oral dose of Compound IA on Day 1, a slight delay in absorption was observed in the case of the encapsulated crystalline tablets under fed conditions. This slower absorption is consistent with the bioavailability results, where no effect of food on t max These findings suggest that the delayed absorption time is due to encapsulation. Renal clearance was determined to be approximately 0.004 L / h (day 1) or 0.008 L / h (day 14), approximately <0.8% of the oral dose. This suggests that direct secretion of Compound IA into urine would not be expected to be a major elimination pathway for this drug in humans.

[0411] Compound IA in the form of a 100 mg crystalline tablet showed a positive food effect (greater exposure in the presence of food), with C in the fed (high-fat, high-calorie meal) state compared to the fasted state. max and AUC increased by 2.05-fold and 1.49-fold, respectively. The median T max for 5 hours, while the suspension reported T max The values ​​are relatively short (0.76-3.0 hours). Compound IA has a very low apparent oral clearance (CLss / F of about 1.0 L / h), which is related to ≤2% of human liver blood flow and a low apparent distribution volume (Vss / F) of about 12.6-23.3 L. Slight drug accumulation of about 1.2 times was observed after QD dosing reached steady state, and slight drug accumulation of 2 times was observed after BID dosing reached steady state, which is consistent with the effective t of about 10 hours determined for crystalline tablets when administered with food. 1 / 2 Consistent.

[0412] Pharmacodynamics

[0413] Nonclinical studies have shown that compound IA blocks the release of IL-1β using a range of NLRP3-dependent activators. This phenomenon is also observed in compounds such as MCC950 (Tapia-Abellan A, Angosto-Bazarra D, Martinez-Banaclocha H, de Torre-Minguela C, Ceron-Carrasco JP, Perez-Sanchez H, Arostegui JI, Pelegrin P. MCC950 closes the active conformation of NLRP3 to an inactive state. [MCC950 closes the active conformation of NLRP3 to an inactive state] Nat Chem Biol [Nature Chemical Biology] 2019; 15: 560-64), which selectively inhibits NLRP3 activation, or in ZYIL1, which shows >90% inhibition of IL-1b in healthy subjects. In contrast, another NLRP3 inhibitor, dapansutrile (OLT1177), only partially reduced the release of IL-1b in healthy subjects and patients with gout attacks.

[0414] In this study, a dose-dependent decrease in IL-1β concentration was observed with increasing single and multiple oral doses of Compound IA. IL-1β production can be mediated by other inflammasomes or inflammasome-independent pathways (Gaidt MM, Hornung V. Alternative inflammasome activation enables IL-1beta release from living cells. Curr Opin Immunol [Current Opinion in Immunology] 2017; 44: 7-13); therefore, inhibitors against IL-1β can lead to unintentional immunosuppression. Therefore, pharmacological inhibitors that specifically target the NLRP3 inflammasome may be a better choice for treating NLRP3-related diseases alone (Zahid A, Li B, Kombe AJK, Jin T, Tao J. Pharmacological Inhibitors of the NLRP3 Inflammasome. Front Immunol 2019; 10: 2538). Safety laboratory findings in 27 subjects were mild non-clinically significant reductions in neutrophil and leukocyte counts. This may be consistent with the PD effect of compound IA caused by inhibition of signaling downstream of NLRP3 (similar to the known effect of anti-IL-1β monoclonal antibody canakinumab) (Dhimolea E. Canakinumab. MAbs 2010; 2: 3-13). Compound IA showed rapid onset of IL-1b inhibition after both single and multiple doses, with a clear dose response throughout the dose range studied and no significant delay in onset, indicating a direct PK / PD relationship. In order to maintain approximately 90% IL-1b inhibition within 24 h, a dose of 25 mg twice daily in the form of a crystalline tablet was selected in a Phase 2a study in knee osteoarthritis.

[0415] The latest report from an early phase 2a clinical trial of Compound IA involving patients with pneumonia and impaired respiratory function caused by COVID-19 infection also showed that Compound IA 50mg BID tablets were well tolerated in this group of patients, and no new safety signals were identified. The results showed that patients who received Compound IA on top of SoC had a trend towards improved response compared to SoC alone. A subanalysis of patients with more severe inflammation (high CRP) but lower corticosteroid doses showed a faster reduction and normalization of inflammatory markers in the COMPOUND IA + SoC group (Madurka I, Vishnevsky A, Soriano JB, Gans SJ, Ore DJS, Rendon A, Ulrik CS, Bhatnagar S, Krishnamurthy S, Mc Harry K, Welte T, Fernandez AA, Mehes B, Meiser K, Gatlik E, Sommer U, Junge G, Rezende E. COMPOUND IA: a new oral NLRP3 inhibitor-tested in an early phase 2a randomised clinical trial in patients with COVID-19 pneumonia and impaired respiratory function. Infection 2022: 1-14).

[0416] In summary, single and multiple oral doses of Compound IA were well tolerated in healthy subjects for up to 14 days with no safety or tolerability issues. The PK profile of Compound IA is compatible with the BID dosing regimen, and the PK / PD data support the dose and formulation selection for further development. The safety and tolerability, PK and PD results suggest that Compound IA has the potential to be an effective oral first-in-class innate immune modulator, which requires further clinical evaluation.

[0417] Example 3

[0418] The following procedures are suitable for testing the activity of NLRP3 inhibitors according to those disclosed herein.

[0419] Procedure 1: IL-1β production in PMA-differentiated THP-1 cells stimulated with gramicidin.

[0420] THP-1 cells are purchased from American Type Culture Collection and are subcultured according to the instructions from the supplier. Before the experiment, cells are cultivated in complete RPMI 1640 (containing 10% heat-inactivated FBS, penicillin (100 units / ml) and streptomycin (100 μg / ml)), and maintained in the logarithmic phase before the experimental setting. Before the experiment, THP-1 was treated with PMA (phorbol 12-myristate 13-acetate) (20ng / ml) for 16-18 hours. The compound is dissolved in dimethyl sulfoxide (DMSO) to produce a 30mM stock solution. On the same day of the experiment, the culture medium is removed and the adherent cells are separated with trypsin for 5 minutes. Then the harvested cells are washed with complete RPMI 1640, spun down, and suspended in RPMI 1640 (containing 2% heat-inactivated FBS, penicillin (100 units / ml) and streptomycin (100 μg / ml)). Cells are plated in 384-well plates at a density of 50,000 cells / well (final assay volume 50 μl). First, the compound is dissolved in assay culture medium to obtain a 5x maximum concentration of 500 μM. Then 10 steps of dilution (1:3) are performed in an assay culture medium containing 1.67% DMSO. 5x compound solution is added to the culture medium to reach the desired final concentration (e.g., 100, 33, 11, 3.7, 1.2, 0.41, 0.14, 0.046, 0.015, 0.0051, 0.0017 μM). The final DMSO concentration is 0.37%. Cells are incubated with the compound for 1 hour, and then stimulated with gramicidin (5 μM) (Enzo) for 2 hours. The plate is then centrifuged for 5 min at 340 g. Cell-free supernatant (40 μL) was collected using a 96-channel PlateMaster (Gilson) and the production of IL-1β was evaluated by HTRF (Xisi Biotechnology). For each experiment, only vehicle controls and CRID3 dose titrations (100-0.0017 μM) were run simultaneously. The data were normalized relative to vehicle-treated samples (equivalent to 0% inhibition) and 100 μM CRID3 (equivalent to 100% inhibition). The compound showed concentration-dependent inhibition of IL-1β production in PMA-differentiated THP-1 cells.

[0421] Procedure 2

[0422] 1. Experimental Procedure

[0423] 1.1 Cell culture

[0424] 1) THP-1 cells were cultured in complete RPMI-1640 medium with 10% FBS at 37°C and 5% CO2.

[0425] 2) By inoculating 3x 10 5 cells / ml, and cells were passaged every 3 days.

[0426] 1.2 Compound preparation

[0427] Serial 3-fold dilutions of compounds were prepared in DMSO in 384-well LDV microplates using the TECAN EVO system to generate a source plate of compounds at 10 concentrations. The highest concentration was 30 mM. Figure 3 depicts the layout of the microplate.

[0428] 1.3 Cell preparation

[0429] 1) Centrifuge THP-1 cells at 350 g for 5 min.

[0430] 2) Resuspend the cells in complete RMPI-1640 medium and count the cells.

[0431] 3) Seed cells in T225 flask (approximately 2.5 x 10 7 Cells were treated with 20 ng / ml PMA (final DMSO concentration < 1%).

[0432] 4) Incubate overnight.

[0433] 1.4 THP-1 stimulation

[0434] 1) Wash adherent THP-1 cells with PBS and detach cells with 4 ml trypsin for use in T225 flasks.

[0435] 2) The cells were centrifuged at 350 g for 5 min, resuspended in RPMI-1640 containing 2% FBS, and counted using trypan blue.

[0436] 3) 50 nl / well of serial dilutions of test compound were transferred to a 384-well plate by Echo; for the high control and first spot of CRID3 (MCC950), 165 nl was transferred and then backfilled to make the DMSO concentration consistent in all wells, the plate layout is as follows.

[0437] 4) Seed 50k cells per well in 40ul RPMI-1640 with 2% FBS in a 384-well plate.

[0438] 5) Incubate at 37°C, 5% CO2 for 1 h.

[0439] 6) Prepare 5x gramicidin, add 10 μl / well to a final concentration of 5 μM, and incubate at 37° C., 5% CO 2 for 2 hours.

[0440] 7) Centrifuge at 350 g for 1 min.

[0441] 8) Pipette 16 μl of supernatant through an apricot and transfer to a white 384 proxiplate. Figure 3 depicts the layout of the plate: HC: 100 μM CRID3 (MCC950) + 5 μM Gramicidin LC: 5 μM Gramicidin.

[0442] 1.5 IL-1β detection

[0443] 1) Homogenize 5x dilution #5 by vortexing and add 1 volume of stock solution to 4 volumes of distilled water.

[0444] 2) Thaw 20x stock solutions of anti-IL1β-cryptate antibody and anti-IL1βXL antibody. Dilute both antibodies to 1x with Assay Buffer #3.

[0445] 3) Premix the two ready-to-use antibody solutions immediately before use.

[0446] 4) Dispense 4ul of premixed anti-IL1β antibody working solution into all wells.

[0447] 5) Seal the plate and incubate overnight at 4°C.

[0448] 6) Use EnVision to read the cell plate and plot the relationship between the reading and the concentration of the test compound to calculate the IC 50 .

[0449] 2. Data Analysis:

[0450] 1. The IC of a compound can be calculated using the following formula 50

[0451] IC 50 Formula

[0452] Inhibition % = 100-100x[HC 平均 -Reading / (HC 平均 -LC 平均 )]

[0453] 2. Use XLfit to fit the normalized data in a dose-response manner and calculate compound concentrations.

[0454] Table 2 shows the biological activity of the compounds in the hTHP-1 assay containing 2% fetal bovine serum: <0.008 μM = "++++++"; ≥0.008 and <0.04 μM = "+++++"; ≥0.04 and <0.2 μM = "++++"; ≥0.2 and <1 μM = "+++"; ≥1 and <5 μM = "++"; ≥5 and <30 μM = "+".

[0455] Compound <![CDATA[hTHP-1IC 50 ]]> I ++++ I A +++++ IB +++

[0456] All publications and patent documents cited herein are incorporated herein by reference, as if each such publication or document is expressly and individually indicated to be incorporated herein by reference. The present invention and its embodiments have been described in detail. However, the scope of the present invention is not intended to be limited to any specific embodiment of the process, manufactured product, material composition, compound, means, method and / or step described in this specification. Various modifications, substitutions and changes can be made to the disclosed materials without departing from the spirit and / or essential features of the present invention. Therefore, it will be easily understood by those of ordinary skill in the art from the present invention that subsequent modifications, substitutions and / or changes that perform substantially the same functions or achieve substantially the same results as the embodiments described herein can be utilized according to such relevant embodiments of the present invention. Therefore, the following claims are intended to cover within their scope modifications, substitutions and changes to the processes, manufactured products, material compositions, compounds, means, methods and / or steps disclosed herein. Unless otherwise specified thereto, the claims should not be understood to be limited to the described order or elements. It should be understood that various changes can be made in form and detail without departing from the scope of the appended claims.

Claims

1. A method of treating an autoinflammatory syndrome in a patient in need thereof, the method comprising administering a therapeutically effective amount of a NLRP3 inhibitor.

2. A method of alleviating symptoms of an autoinflammatory syndrome in a patient in need thereof, the method comprising administering a therapeutically effective amount of a NLRP3 inhibitor.

3. The method of claim 1 or 2, wherein the NLRP3 inhibitor is administered to the subject in a total daily dose of about 50 mg to about 500 mg, optionally about 50 mg to about 200 mg, in a single dose or divided doses.

4. The method of claim 3, wherein the NLRP3 inhibitor is administered to the subject in a single dose or divided doses at a total daily dose of about 100 mg.

5. The method of claim 4, wherein the NLRP3 inhibitor is administered to the subject at a dose of about 100 mg twice a day for three consecutive days and about 100 mg once in the morning of the fourth day.

6. The method of any one of claims 1 to 5, wherein the autoinflammatory syndrome is cryptopyrin-associated periodic syndrome (CAPS), familial cold autoinflammatory syndrome (FCAS), Muckler-Wells syndrome (MWS), neonatal-onset multisystem inflammatory disease / chronic infantile neurocutaneous and articular syndrome (NOMID / CINCA), or familial Mediterranean fever (FMF).

7. The method of any one of claims 1 to 6, wherein the autoinflammatory syndrome is familial cold autoinflammatory syndrome (FCAS).

8. The method of any one of claims 1 to 7, wherein the patient's white blood cell count (WCC) does not increase by more than about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100% after administration of the NLRP3 inhibitor.

9. The method of any one of claims 1 to 7, wherein after administration of the NLRP3 inhibitor, the patient exhibits a lower score of at least 1, at least 2, at least 3 on a scale of 1-10 on a Physician's Gross Assessment Scale following cold exposure.

10. The method of any one of claims 1 to 7, wherein following administration of the NLRP3 inhibitor, the patient exhibits at least a 10%, at least a 20%, at least a 30% lower score on the Physician's Gross Assessment Scale on a scale of 1-100 following cold exposure.

11. The method of any one of claims 1 to 10, wherein the patient's C-reactive protein does not increase by more than about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100% after administration of the NLRP3 inhibitor.

12. The method of any one of claims 1 to 11, wherein the patient does not exhibit more than about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100% increase in IL-1β or IL-18 after administration of the NLRP3 inhibitor.

13. The method of any one of the preceding claims, wherein the NLRP3 inhibitor is administered orally to the subject.

14. The method of any one of the preceding claims, wherein the NLRP3 inhibitor is present in a tablet formulation.

15. The method of any one of the preceding claims, comprising administering at least one additional therapeutic agent.

16. The method of any one of the preceding claims, wherein the NLRP3 inhibitor is Compound 1 or a pharmaceutically acceptable salt thereof:

17. The method of claim 16, wherein Compound I is Compound IA enantiomer or a pharmaceutically acceptable salt thereof:

18. The method of claim 17, wherein Compound IA has an enantiomeric excess of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5%.

19. The method of claim 16, wherein Compound I is an enantiomer of Compound IB or a pharmaceutically acceptable salt thereof:

20. The method of claim 19, wherein Compound IB has an enantiomeric excess of at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5%.

Images