Dosage regimens of NLRP3 inhibitors for the treatment of osteoarthritis

Abstract

The present disclosure relates to the field of pharmaceutical sciences, in particular to the NLRP3 inhibitor for use in the treatment of osteoarthritis.The present disclosure also relates to a pharmaceutical combination comprising an NLRP3 inhibitor or an NLRP3 inhibitor and at least one additional therapeutic agent for use in the treatment of osteoarthritis; a method for treating osteoarthritis, comprising administering an NLRP3 inhibitor or combination; and the use of an NLRP3 inhibitor or combination in the manufacture of a medicament for treating osteoarthritis.

Description

[Technical field]

[0001] The present disclosure relates to the field of pharmaceutical sciences, in particular to the NLRP3 inhibitor for use in the treatment of osteoarthritis.The present disclosure also relates to a pharmaceutical combination comprising an NLRP3 inhibitor or an NLRP3 inhibitor and at least one additional therapeutic agent for use in the treatment of osteoarthritis; a method for treating osteoarthritis, comprising administering an NLRP3 inhibitor or combination; and the use of an NLRP3 inhibitor or combination in the manufacture of a medicament for treating osteoarthritis. [Background technology]

[0002] Osteoarthritis (OA) is the most common joint disorder worldwide and is a serious, chronic, progressive joint disease with no known cure that is associated with an increased risk of premature mortality (Osteoarthritis Research Society International 2016, Paper submitted to US Food and Drug Administration; 01 December; Kluzek et al 2015, Ann Rheum Dis. 75(10):1749-56).

[0003] Clinically, OA is associated with joint pain, swelling and stiffness which can lead to activity limitations, sleep disruption, fatigue, depression, anxiety and ultimately a lack of independence and reduced quality of life (Osteoarthritis Research Society International 2016).

[0004] According to clinical practice guidelines (Bannuru et al. 2019, Cartilage;27:1578-1589; Kolasinski et al. 2020, Arthritis and Rheumatology;72:220-33), non-surgical treatment of OA includes both pharmacological and non-pharmacological modalities (such as patient education, referral to a physiotherapist, exercise, weight loss, walking aids, knee braces, and footwear). All currently available pharmacological treatments for OA provide symptomatic relief by transiently reducing pain but have not been shown to delay the structural damage associated with OA progression. In addition, long-term use of these treatments may be associated with serious adverse effects, including fall-related fractures, drug dependence and / or abuse in patients taking opioids, cardiovascular risk, and upper gastrointestinal bleeding in OA patients taking nonsteroidal anti-inflammatory drugs (Fernandes et al., 2013, Ann Rheum Dis. 72(7):1125-35; McAlindon et al., 2014, Osteoarthritis Cartilage; 22(3):363-88; Nissen et al., 2016, NEJM 375:2519-29; Chan et al. 2017, Lancet 389:2375-82; Soloman et al. 2017, Am. J. Med.; 130:1415-22; Kolasinski et al. 2020).

[0005] Total knee replacement (TKR) is considered when adequate attempts of symptomatic pharmacological treatment fail (Bannuru et al., 2019; Kolasinski et al., 2020). However, not all patients are satisfied with the results or benefit from joint replacement. With increasing longevity and the prevalence of OA (even at a young age), the ever-increasing number of joint replacements will cause an ever-increasing public health burden (Losina and Katz et al., 2012, Arthritis Rheum. 64(2):339-41).

[0006] Proinflammatory cytokines such as interleukin-1β (IL-1β) are crucial mediators of metabolic disorders and hypercatabolism of joint tissues involved in OA (Fraenkel et al 1998, J Rheumatol., 1820-6), making anti-inflammatory therapy an attractive method to combat OA. These inflammatory mediators induce anabolic events, i.e., downregulation of cartilage matrix production by chondrocytes and production of degradative enzymes (MMPs, ADAMTS) by chondrocytes and synovial cells, causing cartilage matrix degradation and loss (van den Bosch 2019, Clin Exp Immunol. 153-166).

[0007] Through the production of IL-1β and IL-18, the NLRP3 inflammasome has been implicated as a key driver of inflammation associated with many chronic inflammatory diseases. Mechanistically, NLRP3 senses a diverse range of danger signals and responds by forming inflammasome protein complexes that drive inflammatory responses. NLRP3 inhibitors have been shown to block IL-1β secretion, IL-18 secretion, and pyroptotic cell death in response to a wide variety of NLRP3-dependent danger signals in vitro and in vivo in mechanistic mouse models.

[0008] There is an unmet need for disease-modifying osteoarthritis drugs (DMOADs) that can slow or stop OA disease progression by inhibiting structural deterioration and improving symptoms. Although many putative agents have been investigated, no drugs have been approved for clinical use as DMOADs (Tonge et al. 2014, Osteoarthritis Cartilage.;22(5):609-21; Karsdal et al. 2016, Osteoarthritis and Cartilage,24:2013-21; Oo et al. 2018, Expert Opin Emerg Drugs Dec;23(4):331-347; Alcaraz et al. 2019, Biochem Pharmacol.165:4-16). Summary of the Invention

[0009] Provided herein are NLRP3 inhibitors that can be used to address unmet medical needs in OA by blocking or reducing the inflammasome response of NLRP3. For example, the NLRP3 inhibitors disclosed herein can be developed as OA drugs to address the inflammatory aspects of the disease and delay / prevent progression to end-stage OA in adults with symptomatic OA, thereby reducing pain, delaying joint damage, and improving function.

[0010] In one aspect, the present invention relates to a method for treating OA, such as OA of the knee, OA of the hand, OA of the hip, OA of the spinal cord, OA of the foot and ankle, by administering to a subject a therapeutically effective amount of an NLRP3 inhibitor, particularly Compound I or a pharma- ceutically acceptable salt thereof.Also described herein are NLRP3 inhibitors, particularly Compound I or a pharma- ceutically acceptable salt thereof, for use in treating OA, such as OA of the knee, OA of the hand, OA of the hip, OA of the spinal cord, OA of the foot and ankle.

[0011] Additionally, as described herein, specific dosing regimens are provided herein for the use of these methods or NLRP3 inhibitors, particularly Compound I or a pharma- ceutically acceptable salt thereof, in the treatment of OA.

[0012] Additionally, pharmaceutical combinations comprising a) Compound I or a pharma- ceutically acceptable salt thereof, and b) at least one additional therapeutic agent, optionally in the presence of a pharma- ceutically acceptable carrier, are described herein for the treatment of OA and pharmaceutical compositions or kits comprising them.

[0013] In certain embodiments, compound I is compound IA.

[0014] Further features and advantages of the described methods and uses will become apparent from the detailed description that follows. [Brief description of the drawings]

[0015] [Figure 1] Schematic overview of the treatment protocol as detailed in Example 1. [Diagram 2] Schematic overview of the study design of the first-in-human (FIH) study as detailed in Example 2. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0016] Definition: In order that this document may be more readily understood, certain terms are first defined. Additional definitions are set forth throughout the document.

[0017] All patents, published patent applications, publications, references and other materials referred to herein are incorporated by reference in their entirety for purposes of disclosure.

[0018] As used herein, the articles "a," "an," and "the" in both the description and the claims should be construed to encompass both the singular and the plural unless otherwise indicated herein or clearly contradicted by context. "Comprising," "having," "including," and "containing" should be construed as open terms (i.e., meaning "including, but not limited to") unless otherwise noted. In addition, whenever "comprising" or another open-ended term is used in an embodiment, it should be understood that the same embodiment may be more narrowly claimed using the intermediate term "consisting essentially of" or the closed term "consisting of."

[0019] The term "or" is used herein to mean, and is used interchangeably with, the term "and / or," unless context clearly indicates otherwise.

[0020] The term "about" or "approximately" as used herein in connection with a reference numerical value and its grammatical equivalents can include the numerical value itself as well as a range of values ​​of ±20% (preferably ±15%, more preferably ±10%, and even more preferably ±5%) from that numerical value. For example, the amount "about 10" includes 10 and any amount from 8 to 12 or 9 to 11. For example, the term "about" in connection with a reference numerical value can also include a range of values ​​of ±10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% from that value. In some cases, a numerical value recited throughout may be "about" that numerical value even without specific reference to the term "about."

[0021] As used herein, the term "baseline" refers to one or more parameters associated with a subject's condition or pathology, e.g., the extent of a disease, or a patient's condition, observed prior to treatment, e.g., prior to administration of a compound, e.g., prior to administration of Compound I, or a pharma- ceutically acceptable salt thereof, optionally in combination with at least one further therapeutic agent, according to the described methods and uses.

[0022] As used herein, the term "administering" in reference to a compound, for example, Compound I, optionally in combination with at least one additional therapeutic agent, is used to refer to delivery of the compound by any delivery route. Such delivery may be, for example, intravenous or oral administration. Such delivery may also be, for example, subcutaneous administration.

[0023] As used herein, the term "substantially" does not exclude "completely", e.g., a composition "substantially free" from Y can be completely free from Y. Where necessary, the term "substantially" may be omitted from the definition.

[0024] As used herein, the term "pharmacologically acceptable" means a non-toxic material that does not substantially interfere with the effectiveness of the biological activity of the active ingredients.

[0025] 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, e.g., non-human primates, sheep, dogs, cats, horses, cows, chickens, amphibians, reptiles, etc. In specific embodiments, the compositions, methods, and uses described herein relate to a human patient or subject.

[0026] As used herein, a subject is "in need of" a treatment if such subject suffers from a condition of interest (i.e., a disease, disorder, or syndrome) and would benefit biologically, medically, or in quality of life from such treatment.

[0027] The terms "treatment", "treating" or "treat" are defined herein as a therapeutic measure for the reduction or amelioration of the progression, severity and / or duration of an undesired physiological change or disorder (e.g., OA, such as OA of the knee), or the amelioration of one or more symptoms (preferably one or more identifiable symptoms) of the disorder resulting from the administration of one or more therapeutic agents. In other embodiments, the terms "treatment", "treating" or "treat" refer to the reduction or stabilization of the progression of a disorder, e.g., OA, either physically, e.g., by the reduction or stabilization of an identifiable symptom, physiologically, e.g., by the reduction or stabilization of a physical parameter, or both. For purposes of this invention, advantageous or desired clinical results include, but are not limited to, relief of symptoms, whether detectable or undetectable, reduction in the extent of disease, a stabilized (i.e., non-worsening) state of disease, delay or slowing of disease progression, remission or palliation of the pathology, and recovery (either partial or total).

[0028] For example, "treatment of OA, such as knee OA, hand OA, hip OA, spinal OA, foot and ankle OA" can refer to ameliorating, reducing or modulating at least one symptom or pathological feature associated with OA; e.g., reducing pain, slowing joint damage, improving function; e.g., slowing the progression of, reducing or halting at least one of, the symptoms or pathological features associated with OA. It can also refer to preventing or slowing one or more of the symptoms described, e.g., slowing, halting or reversing the progression of a disease, condition, disorder, sign or syndrome, and improving clinical outcome.

[0029] "Treating" can also refer to slowing, halting, or reversing the progression of a disease, condition, disorder, sign, or syndrome and improving clinical outcome, e.g., moving from a higher to a lower number on a 5-point scale of clinical signs and symptoms associated with a disease, such as:

[0030] [Table 1]

[0031] As used herein, "excipient" or "pharmaceutically acceptable excipient" means a pharma- ceutically 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 of being compatible with the other components of a pharmaceutical formulation, suitable for use in contact with human and animal tissues or organs, without excessive toxicity, irritation, allergic response, immunogenicity, or other problems or complications, and commensurate with a reasonable risk-benefit ratio. For example, 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.

[0032] As used herein, the term "NLRP3 inhibitor" refers to a compound that directly binds to NLRP3 or inactivates, destabilizes, alters the distribution of, or otherwise inhibits the ability of NLRP3 to induce the production of IL-1β and / or IL-18. Typically, an NLRP3 inhibitor has an IC50 of <1 μM for IL-1β secretion in the hTHP-1 assay with 2% fetal bovine serum as defined herein.

[0033] Preferably, the NLRP3 inhibitor is a compound of Compound I, Compound IA, or Compound IB, or a pharma- ceutically acceptable salt thereof. More preferably, the NLRP3 inhibitor is Compound IA, or a pharma- ceutically acceptable salt thereof.

[0034] As used herein, "compound of formula I" or "compound I" are used interchangeably and refer to a compound having the structure shown below, which is known in the art and can be synthesized using procedures described in WO 2019 / 023147, which is incorporated by reference in its entirety. [ka] Compound I, Compound IA (i.e., (R)-N'-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-2-(2-hydroxypropan-2-yl)thiazole-5-sulfonimidamide) or Compound IB (i.e., (S)-N'-((1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl)-2-(2-hydroxypropan-2-yl)thiazole-5-sulfonimidamide) can be used in crystalline or non-crystalline form, as a solvate, e.g., a hydrate, or in a non-solvated form. [ka]

[0035] Tautomers: The scope of the compounds disclosed herein includes tautomeric forms of the compounds. Thus, by way of example, the moiety [ka] A compound represented as comprising the moiety [ka] It is also intended to include tautomeric forms, including:

[0036] Stereoisomers: Non-limiting exemplary compounds of the formulas described herein include asymmetric sulfur atoms. The present disclosure provides examples of stereoisomeric mixtures (e.g., racemic mixtures of enantiomers). The present disclosure also describes and illustrates methods for separating the individual components of the stereoisomeric mixture (e.g., separating the enantiomers of a racemic mixture). For example, compound I represents a non-racemic mixture of compound IA and compound IB, a racemic mixture of compound IA and compound IB; an enantiomerically pure form of compound IA; or an enantiomerically pure form of compound IB, respectively. As used herein, "compound I" is also intended to include the enantiomeric excess of either compound IA or compound IB. For example, compound IA can 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 can 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%.

[0037] Any chemical formula given herein is also intended to represent unlabeled and isotopically labeled forms of the compound. Isotopically labeled compounds have a structure represented by the chemical formula given herein except that one or more atoms are replaced with 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, e.g., 3 H, 11 C. 13 C. 14 C, and 15 N. Thus, the method of the invention may, for example, 3 H and 14 Isotopes of the above, including radioisotopes such as C, or 2 H and 13 It should be understood that such isotope-labeled compounds may or may not include compounds incorporating one or more of any of the following: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 assays, or in radiotherapy of patients. Isotopically labeled compounds can generally be prepared by conventional techniques known to those of skill in the art, e.g., by substituting an appropriate isotopically labeled reagent for a previously used non-labeled reagent.

[0038] The present invention encompasses embodiments including all pharma- ceutically acceptable salts of compounds useful according to the invention provided herein. As used herein, "pharma- ceutically acceptable salts" refers to derivatives of the disclosed compounds, where the parent compound is modified by converting an existing acid or base moiety into its salt form. Examples of pharma- ceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines; alkali or organic salts of acidic residues such as carboxylic acids; and the like. Pharmaceutically acceptable salts include conventional non-toxic salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. Pharmaceutically acceptable salts can be synthesized from the parent compound containing a basic or acidic moiety by conventional chemical methods. In general, such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or an organic solvent, or in a mixture of the two; generally, non-aqueous media like ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred. Lists of suitable salts can be 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 pharma- ceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines, e.g., the salt can be a hydrochloride salt.

[0039] The phrase "pharmacologically acceptable" as used herein refers to compounds, materials, compositions, and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problems or complications and commensurate with a reasonable risk-benefit ratio.

[0040] Unless otherwise indicated, as used herein, the "dosage" or amount of an NLRP3 inhibitor, such as compound I, or a pharma- ceutically acceptable salt thereof, refers to the amount of the free base or free acid form of the compound. In the case of a salt form of the NLRP3 inhibitor, the actual amount will be adjusted based on the salt form used.

[0041] An "effective amount" refers to an amount sufficient to produce 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 as or different from a prophylactically effective amount, which is the amount necessary to prevent the onset of a disease, condition, disorder, or syndrome or associated symptoms. An effective amount can be administered in one or more administrations, applications, or doses. A "therapeutically effective amount" (i.e., an effective dose) of a therapeutic compound will depend on the therapeutic compound selected. The composition can be administered from once or more times per day to once or more times per week, including less frequent administrations, for example, as described herein. One of skill in the art will appreciate that certain factors, including, but not limited to, the severity of the disease, condition, disorder, or syndrome, previous treatments, the general health and / or age of the subject, and other co-occurring diseases, conditions, disorders, or syndromes, may affect the dosage and timing required to effectively treat a subject. Moreover, treatment of a subject with a therapeutically effective amount of a therapeutic compound described herein may include a single treatment or a series of treatments.

[0042] As used herein, the term "therapeutically effective amount" of a compound described herein refers to an amount of a compound that induces a biological or medical response in a subject, e.g., ameliorates a symptom, reduces a pathology, slows or delays disease progression, or prevents a disease, condition, disorder, sign, or syndrome. In one non-limiting embodiment, the term "therapeutically effective amount" refers to an amount of a compound described herein that, when administered to a subject, is effective to at least partially ameliorate, reduce, or regulate at least one of the symptoms or pathological features associated with OA; e.g., reduce pain, slow joint damage, and improve function (e.g., may refer to slowing the progression of, reducing or stopping at least one of, the symptoms or pathological features associated with OA). It may also refer to preventing or slowing one or more of the symptoms described, e.g., slowing, stopping, or reversing the progression of a disease, condition, disorder, sign, or syndrome, and improving clinical outcome.

[0043] As defined herein, a "combination" refers to either 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, where compound I, or a pharma- ceutically acceptable salt thereof, and one or more additional therapeutic agents may be administered independently simultaneously or separately within time intervals, particularly where these time intervals allow the combination partners to exhibit a synergistic, e.g., synergistic, effect.

[0044] Terms such as "co-administration" or "administration in combination," as used herein, are meant to encompass administration of an additional therapeutic agent to a single subject (e.g., a subject) in need thereof, and are intended to include treatment regimens in which Compound I and the additional therapeutic agent are not necessarily administered by the same route of administration and / or at the same time. Each of the components of the combinations described herein can be administered simultaneously or sequentially, and in any order. Co-administration includes simultaneous, sequential, overlapping, spaced, and / or consecutive administration, and any combination thereof.

[0045] The term "pharmaceutical combination" as used herein means a pharmaceutical composition resulting from the combination (e.g., mixture) of two or more active ingredients and includes both fixed and free combinations of the active ingredients.

[0046] The term "fixed combination" means that the active ingredients are administered to a subject simultaneously in the form of a single entity or dosage.

[0047] The term "free combination" (non-fixed combination) means that the active ingredients as defined herein are administered to a subject as separate entities, either simultaneously, in parallel or sequentially, in any order, without any specific time limitations, such administration providing a therapeutically effective level of the compounds to the subject's body. In particular, as used herein (e.g., in any of the embodiments or claims herein), with respect to combinations including a) compound I and b) at least one additional therapeutic agent, refers to a "non-fixed combination," which can be administered independently, simultaneously or separately, within a time interval.

[0048] "Concurrent administration" means that the active ingredients, as defined herein, are administered on the same day. The active ingredients can be administered simultaneously (in the case of a fixed or free combination) or one at a time (in the case of a free combination).

[0049] The term "sequential administration" may mean that during a period of two or more days of consecutive co-administration, only one of the active ingredients, as defined herein, is administered on any given day.

[0050] "Overlapping administration" means that during a period of two or more days of consecutive co-administration, there may be at least one day of co-administration and at least one day of administration of only one of the active ingredients as defined herein.

[0051] "Consecutive administration" means a period of simultaneous administration without any expiration dates. Consecutive administration may be simultaneous, sequential, or overlapping, as described above.

[0052] The term "dose" refers to a specific amount of a drug to be administered at one time. The dose may be represented, for example, on the product packaging or in a product information pamphlet.

[0053] As used herein, the term "NLRP3" is meant to include, but is not limited to, nucleic acids, polynucleotides, oligonucleotides, sense and antisense polynucleotide strands, complementary sequences, peptides, polypeptides, proteins, homologous and / or orthologous NLRP3 molecules, isoforms, precursors, mutants, variants, derivatives, splice variants, alleles, variants, and active fragments thereof.

[0054] The phrase "means for administering" is used to denote any available device for systemically administering a medication to a patient, including, but not limited to, a dropper, a prefilled syringe, a vial and syringe, a pen injector, an autoinjector, an IV bag, a pump, etc. Such products may be used by patients to self-administer the medication (i.e., administer the medication on their behalf), by a caregiver to administer the medication to the patient, or by a physician or other healthcare professional to administer the medication.

[0055] Treatment methods The present invention provides a method for treating osteoarthritis, comprising administering an NLRP3 inhibitor to a subject. In some embodiments, the osteoarthritis is osteoarthritis of the knee, hand, hip, or spine.

[0056] Treatment with an NLRP3 inhibitor compound according to one of the dosing regimens described herein is expected to slow or halt the progression of osteoarthritis and reduce or eliminate symptoms associated with osteoarthritis compared to treatment with a placebo. In one non-limiting example, treatment may reduce pain based on change from baseline as assessed by Knee Injury and Osteoarthritis Outcome Score (KOOS) or Western Ontario-McMaster Universities Osteoarthritis Index (WOMAC) score. In another embodiment, treatment may reduce stiffness associated with osteoarthritis. In another embodiment, treatment may reduce stiffness associated with osteoarthritis based on change from baseline as assessed by Knee Injury and Osteoarthritis Outcome Score (KOOS) or Western Ontario-McMaster Universities Osteoarthritis Index (WOMAC) score ...KIN) by dynamic contrast enhanced (DCE)-MRI. trans It may result in a reduction in the inflammation level of a joint affected by osteoarthritis, as determined by a change from baseline in the activity level of synovitis measured from . In another non-limiting example, treatment according to one of the dosing regimens described herein may improve or maintain (e.g., prevent further decline) function in an affected joint. In another non-limiting example, treatment according to one of the dosing regimens described herein may prolong the survival of a joint affected by osteoarthritis and / or improve the quality of life of a subject. In yet another non-limiting example, treatment according to the dosing regimens of the present invention may prevent or delay the need for joint replacement surgery. Treatment according to the dosing regimens described below may be continued until such time that the subject no longer receives therapeutic benefit.

[0057] The NLRP3 inhibitor can be administered according to any known administration method. In certain preferred embodiments, the NLRP3 inhibitor is administered via oral administration, for example as a tablet. Other possible administration routes include, for example, intradermal, intramuscular, intravenous, and intraarticular. The NLRP3 inhibitor can also be administered according to any known means for administering therapeutic agents to patients, including, but not limited to, pre-filled syringes, vials and syringes, pen-type injectors, auto-injectors, IV bags, pumps, patch pumps, etc. With such products, patients can self-administer the drug (i.e., administer the drug on their behalf) or a physician can administer the drug.

[0058] Dosage and Administration Regimens The treatment method of the present invention includes administering an NLRP3 inhibitor according to a dosing regimen. In embodiment 1, the dosing regimen includes administering an NLRP3 inhibitor to a subject at a total daily dose of about 10 mg to about 100 mg in a single dose or in divided doses. In embodiment 2, the dosing regimen includes administering an NLRP3 inhibitor to a subject at a total daily dose of about 20 mg to about 50 mg in a single dose or in divided doses. In embodiment 3, the dosing regimen includes administering an NLRP3 inhibitor to a subject at a total daily dose of about 20 mg in a single dose or in divided doses. In embodiment 4, the dosing regimen includes administering an NLRP3 inhibitor to a subject at a total daily dose of about 50 mg in a single dose or in divided doses. In embodiment 5, the dosing regimen includes administering an NLRP3 inhibitor to a subject at about 10 mg twice a day. In embodiment 6, the dosing regimen includes administering an NLRP3 inhibitor to a subject at about 25 mg twice a day. In embodiment 7, the dosing regimen includes administering the NLRP3 inhibitor to the subject at about 10 mg twice daily for about 14 consecutive days. In embodiment 8, the dosing regimen includes administering the NLRP3 inhibitor to the subject at about 25 mg twice daily for about 70 consecutive days. In another embodiment 9, the administration is administered to the subject during or after consumption of a food product. In embodiment 10, the time interval between administration of two subsequent doses is about 10-14 hours. In a preferred embodiment 11, the method of treatment relates to the treatment of knee osteoarthritis. In another preferred embodiment 12, the subject in the method of treatment is a human subject. In another embodiment 13, administration of the NLRP3 inhibitor reduces pain in the joint affected by osteoarthritis as determined by a KOOS score based on the change from baseline. In another embodiment 14, administration of the NLRP3 inhibitor reduces pain in the joint affected by osteoarthritis as determined by a KOOS score based on the change from baseline by dynamic contrast enhanced (DCE)-MRI. In another embodiment 15, administration of the NLRP3 inhibitor reduces pain in the joint affected by osteoarthritis as determined by dynamic contrast enhanced (DCE)-MRI by K transIn another embodiment 15, the level of serum high sensitivity C-reactive protein is reduced by 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% in the subject as determined by change from baseline in the level of synovitis activity measured from. In another embodiment 16, the level of IL-1β or IL-18 is reduced by 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% in the subject as determined by change from baseline. In another embodiment 17, the subject does not exhibit any skin rash. In another embodiment 18, the NLRP3 inhibitor is orally administered to the subject. In another embodiment 19, the NLRP3 inhibitor is included in a tablet formulation. In another embodiment 20, at least one additional therapeutic agent is administered. In another embodiment 21, the NLRP3 inhibitor is compound I, or a pharma- ceutically acceptable salt thereof: [ka]

[0059] In another embodiment 22, the NLRP3 inhibitor is compound IA, or a pharma- ceutically acceptable salt thereof. In another embodiment 23, 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%. In another embodiment 24, the NLRP3 inhibitor is compound IB, or a pharma- ceutically acceptable salt thereof. In another embodiment 25, 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%.

[0060] As will be appreciated by those skilled in the art, the above embodiments of the present invention can be combined with each other.

[0061] Further embodiments of the present invention (embodiments 26.1 to 26.30): 26.1 An NLRP3 inhibitor for use in the treatment of osteoarthritis or use of an NLRP3 inhibitor in the manufacture of a medicament for the treatment of osteoarthritis, wherein the NLRP3 inhibitor is administered to a subject at a total daily dose of about 10 mg to about 100 mg in a single dose or in divided doses. 26.2 NLRP3 inhibitors for use according to embodiment 26.1, administered to a subject in a total daily dose of about 20 mg to about 50 mg in a single dose or in divided doses 26.3 NLRP3 inhibitors for use according to embodiment 26.1 or 26.2, administered to a subject in a total daily dose of about 20 mg in a single dose or in divided doses 26.4 NLRP3 inhibitor for use according to any one of embodiments 26.1 to 26.3, administered to a subject in a total daily dose of about 50 mg in a single dose or in divided doses 26.5 An NLRP3 inhibitor for use according to any one of embodiments 26.1 to 26.4, administered to a subject twice daily at a dose of about 10 mg. 26.6 An NLRP3 inhibitor for use according to any one of embodiments 26.1 to 26.5, administered to a subject twice daily for about 14 consecutive days at a dose of about 10 mg. 26.7 An NLRP3 inhibitor for use according to any one of embodiments 26.1 to 26.6, administered to a subject twice daily at a dose of about 25 mg. 26.8 An NLRP3 inhibitor for use according to any one of embodiments 26.1 to 26.7, administered to a human subject at a dose of about 25 mg twice daily for about 70 consecutive days. 26.9 NLRP3 inhibitor for use according to any one of embodiments 26.1 to 26.8, administered to a subject during or after consumption of a food product 26.10 The NLRP3 inhibitor for use according to any one of embodiments 26.1 to 26.9, wherein there is a time interval of about 10 to 14 hours between administration of two subsequent doses of the NLRP3 inhibitor to the subject. 26.11 The NLRP3 inhibitor for use according to any one of embodiments 26.1 to 26.10, wherein the osteoarthritis is knee osteoarthritis. 26.12 NLRP3 inhibitor for use according to any one of embodiments 26.1 to 26.11, wherein administration of the NLRP3 inhibitor reduces pain in a joint affected by osteoarthritis as determined by a KOOS score based on change from baseline. 26.13 Administration of NLRP3 inhibitors improves K-regulation by dynamic contrast enhanced (DCE)-MRI trans NLRP3 inhibitor for use according to any one of embodiments 26.1 to 26.12, which reduces the level of inflammation in a joint affected by osteoarthritis as measured by a change from baseline in the level of synovitis activity measured from 26.14 The NLRP3 inhibitor for use according to any one of embodiments 26.1 to 26.13, wherein the level of serum high sensitivity C-reactive protein is reduced in the subject by 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%, as determined by change from baseline. 26.15 The NLRP3 inhibitor for use according to any one of embodiments 26.1 to 26.14, wherein the level of IL-1β or IL-18 is reduced in the subject by 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%, as determined by change from baseline. 26.16 The NLRP3 inhibitor for use according to any one of embodiments 26.1 to 26.15, wherein the subject does not exhibit any skin rash. 26.17 NLRP3 inhibitor for use according to any one of embodiments 26.1 to 26.16, administered orally to a subject 26.18 NLRP3 inhibitor for use according to any one of embodiments 26.1 to 26.17, in a tablet formulation 26.19 NLRP3 inhibitor for use according to any one of embodiments 26.1 to 26.18, comprising administering at least one further therapeutic agent 26.20 Compound I, or a pharma- ceutically acceptable salt thereof: [ka] The NLRP3 inhibitor for use according to any one of embodiments 26.1 to 26.19, 26.21 The NLRP3 inhibitor for use according to embodiment 26.20, which is compound IA, or a pharma- ceutically acceptable salt thereof. 26.22 The NLRP3 inhibitor for use according to embodiment 26.21, 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%. 26.23 The NLRP3 inhibitor for use according to embodiment 26.20, which is compound IB, or a pharma- ceutically acceptable salt thereof. 26.24 The NLRP3 inhibitor for use according to embodiment 26.23, 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%. 26.25 The NLRP3 inhibitor for use according to any one of embodiments 26.1 to 26.24, wherein the subject is a human subject. 26.26 A pharmaceutical composition comprising an NLRP3 inhibitor according to any one of embodiments 26.20 to 26.24 for use according to any one of embodiments 26.1 to 26.25.

[0062] Various embodiments of the methods and uses described herein are included below and elsewhere in this document. It will be understood that the features specified in each embodiment can be combined with other specific features to provide further embodiments.

[0063] It is taught herein that the following embodiments relate to the use of any NLRP3 inhibitor, including but not limited to compound I. Preferably, compound I in the following embodiments is compound IA (i.e., R enantiomer) with 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%. Preferably, compound IA is at an enantiomeric excess of at least 90%. More preferably, compound IA is at an enantiomeric excess of at least 95%.

[0064] In some embodiments, provided herein is a pharmaceutical composition comprising Compound I or a pharma- ceutically acceptable salt thereof and at least one pharma- ceutically acceptable excipient. In certain embodiments, the pharmaceutical composition is a tablet. In more particular embodiments, the pharmaceutical composition is administered as a whole or crushed tablet. In some embodiments, the pharmaceutical composition comprises 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 in each unit dose.

[0065] Provided herein is a pharmaceutical composition comprising Compound I, or a pharma- ceutically acceptable salt thereof, for use in any of the embodiments described herein.

[0066] In any of the embodiments described herein, compound I, or a pharma- ceutically acceptable salt thereof, is orally administered to a subject in need thereof.In some embodiments, compound I, or a pharma- ceutically acceptable salt thereof, is in the form of a tablet that is administered either whole or com- pounded, i.e., crushed before administration.In certain embodiments, compound I can be administered via a nasogastric tube, for example, when a patient is unable to swallow.

[0067] Synthesis of Compound I Compounds I, IA and IB were synthesized according to the synthesis defined in WO2019 / 023147, e.g., 4, 5 and 6, and as detailed below. However, the compounds can be assembled in various ways to build the final molecules using related reaction procedures in a modular fashion allowing for different reaction sequences and / or different reagents.

[0068] Reaction progress was often monitored by TLC or LC-MS. Product identity was often confirmed by LC-MS. LC-MS was recorded using the following methods: Method A: Shim-pack XR-ODS, C18, 3 x 50 mm, 2.5 um column, 1.0 uL injection, 1.5 mL / min flow rate, scan range 90-900 amu, UV range 190-400 nm, gradient 5-100% (1.1 min), 100% (0.6 min) with ACN (0.05% TFA) and water (0.05% TFA), 2 min total run time.

[0069] The final target was purified by preparative HPLC, which was performed using the following method: Method B: Preparative HPLC: Column, XBridge Shield RP18 OBD (19×250 mm, 10 um); Mobile phase, water (10 mmol / L NH4HCO3) and ACN, UV detection 254 / 210 nm

[0070] NMR was recorded on a BRUKER NMR 300.03 MHz, DUL-CH, ULTRASHIELD™ 300, AVANCE II 300B-ACS™ 120 or a BRUKER NMR 400.13 MHz, BBFO, ULTRASHIELD™ 400, AVANCE III 400, B-ACS™ 120 or a BRUKER AC250 NMR instrument with TMS as the reference measured in ppm (parts per million). [ka]

[0071] Compound I: [ka] Compound I: N'-(1,2,3,5,6,7-hexahydro-s-indacen-4-ylcarbamoyl)-2-(2-hydroxypropan-2-yl)thiazole-5-sulfonimidamide Step 1: N-(tert-butyldimethylsilyl)-N'-(1,2,3,5,6,7-hexahydro-s-indacen-4-ylcarbamoyl)-2-(2-hydroxypropan-2-yl)thiazole-5-sulfonimidamide: A 50 mL round bottom flask was charged with a solution of N'-(tert-butyldimethylsilyl)-2-(2-hydroxypropan-2-yl)thiazole-5-sulfonimidamide (Intermediate I) (336 mg, 1.0 mmol) in THF (10 mL). To this solution, NaH (60% wt, 80 mg, 2.0 mmol) was added portionwise at 0° C. The solution was stirred at 0° C. for 15 min, followed by the dropwise addition of a solution of 4-isocyanato-1,2,3,5,6,7-hexahydro-s-indacene (209 mg, 1.1 mmol) in THF (5 mL) with stirring at RT. The resulting solution was stirred at RT for 12 h. The reaction was then quenched by the addition of 10 mL of NH4Cl (sat.). The resulting solution was extracted with 3×10 mL of DCM and the combined organic layers were concentrated under vacuum. This gave 535 mg (crude) of the title compound as a brown oil. MS-ESI: 535.0 (M+1).

[0072] Step 2: N'-(1,2,3,5,6,7-hexahydro-s-indacen-4-ylcarbamoyl)-2-(2-hydroxypropan-2-yl)thiazole-5-sulfonimidamide: A 50 mL round bottom flask was charged with a solution of N-(tert-butyldimethylsilyl)-N'-(1,2,3,5,6,7-hexahydro-s-indacen-4-ylcarbamoyl)-2-(2-hydroxypropan-2-yl)thiazole-5-sulfonimidamide (535 mg, crude, 1.0 mmol). To this solution, HF / Py (70% wt, 143 mg, 5.0 mmol) was added dropwise at 0 °C. The solution was stirred at RT for 4 h. The reaction was then quenched by the addition of 10 mL of water. The resulting solution was extracted with 3 × 10 mL of 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%-60% within 10 min). This afforded 189 mg (45%, 2 steps) of compound I as a white solid. Compound I:MS-ESI:421.0(M+1).1H NMR(400MHz,DMSO-d6)δ 8.46(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).

[0073] Compound IA and Compound IB: [ka] Compounds IA and IB: (R) and (S)-N'-(1,2,3,5,6,7-hexahydro-s-indacen-4-ylcarbamoyl)-2-(2-hydroxypropan-2-yl)thiazole-5-sulfonimidamide: Step 3: Chiral separation The product of compound I (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 (20% EtOH held over 16 min); flow rate, 20 mL / min; detector, UV 254 / 220 nm. This gave 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. Compound IB:MS-ESI:421.0(M+1).1H NMR(400MHz,DMSO-d6)δ 8.43(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). Compound IA:MS-ESI:421.0(M+1).1H NMR(400MHz,DMSO-d6)δ 8.41(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).

[0074] Intermediate I of Scheme 1 was synthesized according to the synthesis described in WO 2019 / 023147 and as presented in Scheme 2 below. [ka]

[0075] Intermediate I: [ka] N'-(tert-butyldimethylsilyl)-2-(2-hydroxypropan-2-yl)thiazole-5-sulfonimidamide: Step 1: 2-(2-methyl-1,3-dioxolan-2-yl)thiazole: A 500 mL round bottom flask was charged with a solution of 1-(thiazol-2-yl)ethanone (20 g, 157.0 mmol) and ethane-1,2-diol (19.5 g, 314 mmol) in toluene (300 mL). To the solution was added TsOH (2.7 g, 15.7 mmol). The resulting solution was refluxed overnight, during which water separated from the solution. The resulting solution was diluted with 200 mL of water and extracted with 2×100 mL of ethyl acetate. The organic layers were combined, dried over anhydrous Na2SO4, and then concentrated under vacuum. This afforded 26.6 g (99%) of the title compound as a pale yellow oil. MS-ESI: 172.0 (M+1).

[0076] Step 2: 2-(2-methyl-1,3-dioxolan-2-yl)thiazole-5-sulfonamide: A solution of 2-(2-methyl-1,3-dioxolan-2-yl)thiazole (14 g, 81.6 mmol) in THF (200 mL) was placed in a 500 mL 3-neck round bottom flask purged with nitrogen and maintained under nitrogen. This was followed by dropwise addition of n-BuLi (2.5 M in THF, 35.2 mL, 88.0 mmol) with stirring at -78 °C. The resulting solution was stirred at -78 °C for 0.5 h and then SO2 was introduced to the above reaction mixture. The reaction was gradually warmed to RT and then NCS (12.8 g, 95.86 mmol) was added. The resulting solution was stirred at RT for 1 h. The solids were filtered. The resulting filtrate was concentrated under vacuum and then diluted with DCM (160 mL). To it was added a saturated solution of ammonia in DCM (300 mL). The resulting solution was stirred at RT for 3 h 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 afforded 12.5 g (61%) of the title compound as a yellow solid. MS-ESI: 251.0 (M+1).

[0077] Step 3: 2-Acetylthiazole-5-sulfonamide: A 250 mL round bottom flask was charged with a solution of 2-(2-methyl-1,3-dioxolan-2-yl)thiazole-5-sulfonamide (12.5 g, 50.0 mmol) in THF (125 mL). To it was added aq. HCl (4N, 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 afforded 9.3 g (90%) of the title compound as a yellow solid. MS-ESI: 207.0 (M+1).

[0078] In steps 4-6, intermediate I was obtained from compound Id using the same specific procedure for converting compound Z to compound Y shown in Scheme 3. MS-ESI: 336.1 (M+1). [ka]

[0079] Compound Y: [ka] N'-(tert-butyldimethylsilyl)-5-(2-hydroxypropan-2-yl)thiazole-2-sulfonimidamide: Step 1: Methyl 2-mercaptothiazole-5-carboxylate: A 2 L round bottom flask was charged with methyl 2-bromothiazole-5-carboxylate (100 g, 450 mmol), EtOH (1000 mL), and sodium hydrogen sulfide (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 (1N). The solid was collected by filtration. This gave 63.2 g (80%) of the title compound as a pale yellow solid. MS-ESI: 176.0 (M+1).

[0080] Step 2: Methyl 2-(chlorosulfonyl)thiazole-5-carboxylate: A 1 L round bottom flask was charged with methyl 2-mercaptothiazole-5-carboxylate (30 g, 170 mmol) and acetic acid (300 mL). This was followed by the addition of sodium hypochlorite (300 mL, 8%-10% wt.) portionwise at 0° C. The resulting solution was stirred at RT for 2 h and then diluted with 500 mL of water. The solution was extracted with 3×300 mL of DCM and the combined organic layers were washed with 2×300 mL of brine and dried over anhydrous Na2SO4. The crude product as a yellow solution in DCM was used in the next step.

[0081] Step 3: Methyl 2-sulfamoylthiazole-5-carboxylate: A 2 L round bottom flask was charged with methyl 2-(chlorosulfonyl)thiazole-5-carboxylate as a crude solution in DCM (900 mL). To the solution was introduced NH3(g) for 20 min below 0 °C. The resulting solution was stirred at RT for 1 h and 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 afforded 23 g (75%, 2 steps) of the title compound as a white solid. MS-ESI: 223.0 (M+1).

[0082] Step 4: 5-(2-hydroxypropan-2-yl)thiazole-2-sulfonamide: A solution of methyl 2-sulfamoylthiazole-5-carboxylate (15 g, 67.5 mmol) in THF (150 mL) was placed in a 500 mL round bottom flask purged with nitrogen and maintained under nitrogen. This was followed by dropwise addition of MeMgBr / THF (3 M, 90 mL) with stirring at 0 °C. The resulting solution was stirred at RT for 14 h and then quenched by addition of 100 mL of NH4Cl (sat.). The resulting solution was extracted with 3 × 150 mL of DCM. 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 ethyl acetate / petroleum ether (1:5 to 1:3). This afforded 11.5 g (78%) of the title compound as a white solid. MS-ESI: 223.0 (M+1), 221.0 (M-1) (positive and negative ion modes, respectively).

[0083] Step 5: N-(tert-butyldimethylsilyl)-5-(2-hydroxypropan-2-yl)thiazole-2-sulfonamide: A 250 mL 3-neck round bottom flask purged with nitrogen and maintained under nitrogen was charged with a solution of 5-(2-hydroxypropan-2-yl)thiazole-2-sulfonamide (5 g, 22.5 mmol) in THF (100 mL). To it was then added NaH (60% wt, 1.8 g, 45.0 mmol) in an ice / water bath in small portions. After stirring in the water / ice bath for 20 min, this was followed by dropwise addition of a solution of TBSCl (4.1 g, 27.2 mmol) in THF (10 mL) with stirring at 0 °C. The resulting solution was stirred at RT for 4 h. The reaction was quenched with saturated NH4Cl (100 mL). The resulting solution was extracted with 3 x 100 mL of ethyl acetate and the combined organic layers were dried over Na2SO4 and concentrated under vacuum. The crude solid was washed with ethyl acetate / hexane (1:5) (2 x 100 mL). This gave 6.81 g (90%) of the title compound as a yellow solid. MS-ESI: 337.1 (M+1), 335.1 (M-1) (positive and negative ion modes, respectively).

[0084] Step 6: N'-(tert-butyldimethylsilyl)-5-(2-hydroxypropan-2-yl)thiazole-2-sulfonimidamide: A solution of PPh3Cl2 (3 g, 9.0 mmol) in CHCl3 (100 mL) was placed in a 100 mL 3-neck round bottom flask purged with nitrogen and maintained under nitrogen. This was followed by dropwise addition of DIEA (1.54 g, 11.9 mmol) with stirring at RT. The resulting solution was stirred at RT for 10 min. This was followed by dropwise addition of a solution of N-(tert-butyldimethylsilyl)-5-(2-hydroxypropan-2-yl)thiazole-2-sulfonamide (2.0 g, 5.9 mmol) in CHCl3 (30 mL) with stirring in an ice / water bath. The resulting solution was stirred in the ice / water bath for 30 min. To it was introduced NH3(g) for 15 min below 0 °C. The resulting solution was stirred at RT for 20 min. The solid was filtered, the filtrate was concentrated, and the residue was dissolved in 300 mL of ethyl acetate. The solution was washed with brine (2 x 100 mL), dried over Na2SO4, and concentrated under vacuum. The crude solid was washed with CHCl3 (100 mL). 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 original washed solid and the solid from the silica gel purification were combined. This afforded 1.2 g (60%) of the title compound as a white solid. MS-ESI: 336.1 (M+1).1H-NMR (300 MHz, DMSO-d6) δ 7.66 (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).

[0085] The following abbreviations have the meanings indicated: ACN = acetonitrile BTC = trichloromethyl chloroformate Boc = t-butyloxycarbonyl Davephos = 2-dicyclohexylphosphino-2'-(N,N-dimethylamino)biphenyl DCM = dichloromethane DEA = diethylamine DMF = N,N-dimethylformamide DMSO = dimethyl sulfoxide DIEA = N,N-Diisopropylethylamine DPPA = diphenylphosphoryl azide dppf = 1,1'-bis(diphenylphosphino)ferrocene EtOH = ethanol HATU = 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate Hex = Hexane HPLC = High Performance Liquid Chromatography LC-MS = Liquid Chromatography-Mass Spectrometry LiHMDS = lithium bis(trimethylsilyl)amide LDA = lithium diisopropylamide M = moles / L Me = methyl MeOH = methanol MSA = methanesulfonic acid NBS = N-bromosuccinimide NCS = N-chlorosuccinimide NMR=nuclear magnetic resonance Pd(dppf)Cl2 = dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium Ph = phenyl PPh3Cl2 = Dichlorotriphenylphosphorane Py = pyridine RT=room temperature Rt=retention time Rf = Delay factor Sat.=saturated TBAF = Tetrabutylammonium fluoride TBS = tert-butyldimethylsilyl TBSCl = tert-butyldimethylsilyl chloride TBDPSCl = tert-butyldiphenylsilyl chloride TEA = triethylamine TFA = trifluoroacetic acid THF = tetrahydrofuran TLC = Thin Layer Chromatography TsOH = 4-methylbenzenesulfonic acid UV=ultraviolet light bid=2 times a day WCC=white blood cell count EP=Endpoint y=year y / n=Yes / No EXAMPLES

[0086] The following examples illustrate the methods and uses described herein. However, they are in no way intended to limit the scope of the described methods and uses. Other variations of the embodiments will be readily apparent to those skilled in the art and are encompassed by the appended claims.

[0087] Example 1: Clinical Trials with Compound IA The starting dose of Compound IA in participants enrolled in this study will be set as a single-step dose escalation to 10 mg twice daily (bid) for 2 weeks, followed by 25 mg bid for 10 weeks, for a total treatment period of 12 weeks to evaluate tolerability. The initial dose level was selected primarily against data from the first-in-human (FIH) study (Example 2), where skin rash has been observed in some participants receiving 30, 100, and 200 mg Compound IA once daily. The mechanism of skin rash is still unknown, but is being explored as it could potentially mitigate the risk when administered bid. In addition to evaluating the tolerability of Compound IA, a 2-week run-in period at the lower 10 mg dose bid will also provide information on the extent of peripheral PD marker inhibition.

[0088] A randomized, two-arm, placebo-controlled, participant- and investigator-blinded, phase 2 study investigates the efficacy, safety, and tolerability of Compound IA in patients with symptomatic knee osteoarthritis. Figure 1 is a schematic overview of the treatment protocol with Compound IA (i.e., the R enantiomer of Compound I). In the first arm of the clinical trial, 10 mg of Compound IA is administered orally twice daily for 13 consecutive days, and 10 mg in the morning on the 14th day, i.e., the total dose in the first arm is 270 mg. The second arm of the study begins immediately after the first arm: 25 mg is administered in the evening on the 14th day, then 25 mg is administered twice daily for 69 days, and in the morning on the 84th day, i.e., the total dose in the second arm is 3500 mg.

[0089] This Phase 2 study should evaluate the safety and tolerability of Compound IA in participants with symptomatic knee OA and determine the efficacy of Compound IA in reducing knee pain as evidenced by the KOOS (Knee Injury and Osteoarthritis Outcome Score).

[0090] The primary endpoint (EP) was change from baseline in the Knee Injury and Osteoarthritis Outcome Score (KOOS) pain subscale at 12 weeks.

[0091] The secondary objectives of the study were: -To evaluate the safety and tolerability of Compound IA; -To evaluate the efficacy of compound IA on inflammatory joint structural features; - To evaluate the effect of Compound IA on systemic inflammatory conditions; - To evaluate the pharmacokinetics of Compound IA in plasma; -To evaluate the effectiveness of Compound IA in improving participants' reports of knee symptoms and related problems over time; - To evaluate the effectiveness of Compound IA in improving participants' reported knee symptoms

[0092] The secondary endpoints (EPs) were: safety endpoints (including vital signs, ECG parameters, safety laboratory assessments and adverse events); K by DCE-MRI at 12 weeks trans change from baseline in synovitis activity level measured from; Changes from baseline in serum high-sensitivity C-reactive protein levels and absolute neutrophil counts at weeks 2, 4, 8, and 12; Quantifying the concentration of Compound IA at various time points (weeks 2 and 12) and changes in plasma samples to derive plasma PK parameters (including but not limited to Cmax, AUClast, AUC0-12h, and Ctrough); change from baseline in KOOS subscales (other symptoms, function in daily living, function in sports and recreation, knee-related quality of life) at weeks 2, 4, 8, and 12; Change in Numeric Rating Scale (NRS) for Pain from Baseline to Weeks 2, 4, 8, and 12

[0093] [Table 2]

[0094] [Table 3]

[0095] Test Design: The study will use a randomized, two-arm, parallel-group, participant- and investigator-blinded, placebo-controlled design to evaluate the safety and tolerability of oral Compound IA in approximately 108 participants with symptomatic inflammatory knee OA after 12 weeks of treatment and determine the efficacy of Compound IA, as evidenced by reduction in knee pain by KOOS (Knee Injury and Osteoarthritis Outcome Score).

[0096] The study will consist of a screening period of up to 45 days used to assess eligibility and reduce unauthorized medications from participants. At the Day 1 visit, eligible participants will be randomized to one of the treatment arms. Eligible participants will enter the treatment period, beginning with a 2-week titration period where they take Compound IA or placebo orally at 10 mg twice daily for 14 consecutive days, followed by a 10-week treatment period where they take Compound IA or placebo orally at 25 mg twice daily. The end of study visit will occur 15 days after the last dose, and a post-study safety contact will occur 30 days after the last dose. The total study time from screening to end of study is expected to be up to 19 weeks.

[0097] Assessments to address the primary objective will be conducted at the end of the treatment period (week 12).

[0098] The study includes three periods: -Screening Period: The screening period consists of two visits, the screening visit and the baseline visit. Treatment period: The treatment period consisted of 5 visits. - Follow-up Period: Patients will be followed up at the end of the study visit, approximately 15 days after the last dose. In addition, a safety follow-up call will be conducted approximately 30 days after the last dose.

[0099] Treatment duration: The treatment period consisted of five visits: -Treatment Initiation Visit (Day 1): Participants who meet all inclusion criteria and no exclusion criteria will be enrolled and begin taking Compound IA or matching placebo tablets at a total daily dose of 20 mg (10 mg bid) twice daily for 14 consecutive days (last dose on Day 14, administered in the morning). The first dose will be either Compound IA 10 mg or placebo, and participants will be dispensed study medication and prepared for ongoing treatment at home. Participants may reside the evening before the scheduled visit at the discretion of the participant and investigator for their convenience and logistical aspects. -Participants will be evaluated on Day 14 as outlined in the evaluation schedule. Provided treatment is well tolerated based on the investigator's judgment and guidelines, they will begin taking Compound IA or matching placebo tablets (bid) at a total daily dose of 50 mg (25 mg bid) starting on Day 14 (evening doses only) for 10 weeks. The final dose will be administered on Day 84 (morning doses only). -Participants will undergo assessments on Days 28, 56, and 84. Site staff will contact participants by phone at least once between bimonthly visits to remind them of their study medication intake.

[0100] Follow-up period: Participants will be followed up at the end of study visit, approximately 15 days after the last dose (Day 99), and for end of study evaluations. A safety follow-up call will be conducted approximately 30 days after the last dose (Day 114) to document any potential safety events.

[0101] Inclusion Criteria: Participants eligible for inclusion in this study must meet all of the following criteria: 1. Written informed consent must be obtained prior to conducting any evaluations. 2. Able to communicate well with the investigator and understand and comply with the requirements of the study. 3. Male and female participants >=50 and <=80 years of age on the date of signing the informed consent 4. Participants must weigh at least 50 kg at screening to participate in the study, and must be between 18 and 35 kg / m 2 BMI = weight (kg) / [height (m)]2 5. High-sensitivity C-reactive protein (hsCRP) >= 1.8mg / L at screening 6. Symptomatic OA with pain (Numerical Rating Scale [NRS] 5-9, inclusive) in the target knee for most days within the last 3 months prior to screening. At screening, patients will be given a diary to record pain and analgesic use. Participants must have PRO-reported NRS pain ≥5 to ≤9 at screening and baseline according to the diary entries for at least 5 of the 7 days prior to baseline. 7. The primary cause of pain was due to OA in the target knee based on a Widespread Pain Index (WPI) score <= 4 at screening 8. KOOS pain subscale score <= 60 in the index knee at screening and baseline 9. Radiological disease: At screening, radiologically confirmed knee osteoarthritis with K&L grade 2 or 3 in the target knee according to the OARSI atlas Active synovial inflammation at screening, defined as either moderate (score 9–12) or severe (score >= 13) based on contrast-enhanced MRI (CE-MRI) of the whole knee to detect synovitis from 10.11 sites [Guermazi et al 2011] 11. Diagnosis of primary tibiofemoral knee OA by standard American College of Rheumatology (ACR) clinical and radiographic criteria at screening 12. Current use of analgesic therapy to control local pain in the target knee: Patients taking paracetamol / acetaminophen, including combination medications containing low-dose opioids, may continue to do so in accordance with the package insert / physician's instructions. Patients taking any other analgesic including NSAIDs and selective cyclooxygenase 2 inhibitors, but not topical NSAIDs or steroids, must be willing to switch to paracetamol / acetaminophen, including combination medications containing low-dose opioids, at screening, for any pain indication, including knee pain, per package insert / physician's recommendation. NSAIDs are permitted exclusively as rescue medication, but must not be used within 48 hours or 5 half-lives, whichever is longer, prior to any PRO assessment. Patients taking glucosamine or chondroitin should discontinue these at screening

[0102] Key exclusion criteria: Participants who meet any of the following criteria are ineligible for inclusion in this study: 1. At screening, total WBC count <3,000 / μL, absolute peripheral blood neutrophil count (ANC) <1,000 / μL, hemoglobin <8.5g / dL (85g / L), or platelet count <100,000 / μL 2. Known autoimmune disease with inflammatory arthritis (including but not limited to rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis, systemic lupus erythematosus), crystalloid arthritis (gout, pseudogout-related arthritis), active acute or chronic infection of the knee joint or history of infection, Lyme disease involving the knee, reactive arthritis, generalized cartilage damage, moderate to severe fibromyalgia (Wide Pain Index, WPI, >4 out of 19), or known systemic connective tissue disease 4. Arthropathy-related disorders of metabolic or genetic origin 5. Participant has an unstable target knee joint, inadequate reconstruction of knee hardware or ligaments based on medical history at screening and physical examination by the investigator 6. Participant has symptomatic localized patellofemoral pain in the index knee according to investigator examination at screening 7. Use of electrotherapy, acupuncture, and / or chiropractic treatment for knee OA within 4 weeks prior to screening 8. Any known active infection, including skin or knee infections or infections that may impair the immune system, e.g., HIV or chronic Hepatitis B or C infection. Specific to COVID-19: It is highly recommended that PCR or antigen testing for COVID-19 be completed within 1 week prior to first dose. If testing is performed, a negative test result is required prior to enrollment in the study. Additional testing may be performed at the investigator's discretion. COVID-19 testing should be completed via nasal or throat swab. If testing is not performed, investigators must document in the source documentation their discussion with the participant regarding the rationale for testing and not testing. This requirement may be disregarded if the pandemic is declared over by the country in which the site is located and may be reinstated if the pandemic reoccurs. 9. Any diagnosed psychiatric condition, including but not limited to, a history of mania, bipolar disorder, psychosis, schizophrenia, or schizoaffective disorder, depression, or anxiety, which, in the judgment of the investigator, may jeopardize the participant's safety or compliance with study procedures. 10. History of lymphoproliferative disorder or any known malignancy within 5 years of screening or history of malignancy of any organ system (except basal cell carcinoma or actinic keratosis that has been treated and has no evidence of recurrence within 3 months prior to screening, or carcinoma in situ of the cervix or noninvasive malignant colon polyp that has been removed) 11. Symptomatic hip OA or hip prosthesis recently implanted (within 1 year prior to screening) or predicted within the study period (on either side) 12. Other pathology affecting the knee based on clinical or imaging assessment, including subchondral fragility fracture, fracture (acute or subacute within 6 months prior to screening) or bone contusion, osteonecrosis, osteochondral lesion, malignant bone marrow infiltration, solid tumor, meniscus protrusion of >50% and / or softened meniscus and / or patellofemoral dysplasia 13. Unstable target knee (including but not limited to post-traumatic or congenital laxity) or inadequate ligament reconstruction based on medical history and / or physical examination by the investigator 14. Prohibited Substance Use: Any local ia treatment to the knee, including but not limited to viscosupplementation and corticosteroids within 12 weeks prior to Day 1; chronic treatment (>14 days) with oral corticosteroids >5 mg / day within 4 weeks prior to Day 1; oral glucosamine, chondroitin sulfate, or any functional food with potential activity on cartilage repair within 2 weeks prior to Day 1; systemic nonsteroidal anti-inflammatory drugs (NSAIDs) or selective cyclooxygenase-2 inhibitors within 48 hours or 5 half-lives, whichever is longer, of PRO assessment; any other immunomodulatory agent, or treatment that cannot be discontinued or switched to a different medication within 28 days or 5 half-lives (whichever is longer if required by local regulations) of screening or until expected PD effects have returned to baseline. 15. Severe malalignment (either varus or valgus) of greater than 7.5 degrees in the target knee as measured with x-ray at screening 16. Participants who are unable or unwilling to undergo MRI or have contraindications to MRI (e.g., metal implants, metal foreign bodies, pacemakers, defibrillators) or to the use of gadolinium-based agents, e.g., previous severe allergic / anaphylactoid reactions to gadolinium-based contrast agents; severe renal disease (at screening and baseline, eGFR < 60 mL / min calculated using the CKD-EPI formula [https: / / www.kidney.org / professionals / KDOQI / gfr_calculator] or protein >= 2+ on urine dipstick test), or acute deterioration of renal function 17. Moderate to severe pain in the contralateral knee for most days within the last 3 months prior to screening, as determined by the patient 18. History of knee replacement or planned knee replacement (partial or total) in either knee. Any other previous surgical procedure in the target knee, including osteochondral grafting, microfracture, meniscectomy >50% or osteotomy. Arthroscopy or lavage of the target knee within 6 months prior to screening or scheduled during the study. 19.Females of childbearing potential, defined as any female who is physiologically capable of becoming pregnant unless a highly effective method of contraception is used during treatment with the study drug and for 15 days after discontinuation 20. Pregnant or nursing (breastfeeding) women 21. History or current diagnosis of ECG abnormalities indicating a significant safety risk in participants participating in studies such as: - Concomitant clinically significant cardiac arrhythmias not involving a pacemaker, e.g., sustained ventricular tachycardia and clinically significant second or third degree AV block - History of familial long QT syndrome or known family history of polymorphic ventricular tachycardia 22. History of drug abuse or unhealthy alcohol use within 12 months prior to expected first dose, or evidence of abuse as demonstrated by laboratory assays performed at the screening visit. 23. History of hypersensitivity to any of the study treatments or excipients or to drugs of a similar chemical class 24. Use of other investigational drugs within 5 half-lives of enrollment or until expected pharmacodynamic effects return to baseline, whichever is longer. 25. Primary knee osteoarthritis plus primary osteoarthritis of the spine / hand / shoulder / hip / foot / other must have been present for at least 3 months prior to screening and diagnosis and symptoms must be documented as determined by the investigator. 26. Secondary osteoarthritis with history or any evidence in the potential target joint of the following diseases: septic arthritis, inflammatory joint disease, gout, recurrent episodes of pseudogout, Paget's disease of bone, articular fractures, ochronosis, acromegaly, hemochromatosis, Wilson's disease, primary osteochondromatosis, hereditary disorders, collagen gene mutations. 27. Participant is taking concomitant medications known to be strong or moderate inducers of the cytochrome CYP2C9 enzyme and / or strong inhibitors of CYP2C9 and / or strong inducers of CYP3A, and treatment cannot be discontinued or switched to a different medication within 5 half-lives or 1 week (whichever is longer) prior to Day 1 and for the duration of the study. 28. History of clinically significant liver disease or injury as indicated by abnormal liver function tests (as defined below), including but not limited to SGOT (AST), SGPT (ALT), alkaline phosphatase, serum bilirubin, albumin, and prothrombin time. The investigator must be guided by the following criteria: - Any single parameter must not exceed 2 × upper limit of normal (ULN) 29.CYP2C9 * CYP2C9, defined as homozygous carriers of 3 alleles * 3 / * Participants with 3 genotypes 30. Onset of symptoms or diagnosis of primary osteoarthritis other than in the knee joint <3 months prior to screening 31. Live vaccine within 4 weeks of Day 1 (i.e., first dose of Compound IA) 32. Known history of renal disease, including nephrolithiasis

[0103] Efficacy assessment: The efficacy assessments described in this section will be assessed in all participants in both treatment arms. Pain (primary endpoint) will be assessed by patient-reported outcomes (PROs). Pharmacodynamic samples will be collected.

[0104] Synovitis (secondary endpoint), articular cartilage volume / thickness and exudate volume (exploratory endpoints) will be assessed from MRI.

[0105] Pharmacodynamic (PD) samples will be obtained and evaluated at all dose levels in all participants, including the placebo group.

[0106] Patient-reported outcomes (PROs): Participants will be given a PRO scale to be completed at the scheduled visit before any other clinical assessments are performed. The questionnaire should be completed in the language most familiar to the participant. Participants should be given sufficient space and time to complete the PRO scale. Participants' refusal to complete any part of the PRO scale should be documented in the case report / record form (CRF). Study staff should review collected PRO scales for completion and ask participants to complete any missing answers. Completed PROs, including any unsolicited comments written by participants, must be reviewed and assessed by the investigator for answers that may include potential AEs or SAEs before any clinical trial is conducted. If an AE or SAE is identified, the study investigator should not prompt participants to change the answers reported in the completed questionnaire.

[0107] Knee Injury and Osteoarthritis Outcome Score (KOOS): Knee-related pain is assessed as the primary endpoint using the Knee Injury and Osteoarthritis Outcome Score (KOOS) scale collected at regular intervals (Roos EM, Davis AM (2012) Recommendations for publication of cross-cultural validation studies of patient-reported outcomes (PROs) in Osteoarthritis and Cartilage. Osteoarthritis Cartilage. p. 4-5.). The KOOS includes 42 items broadly divided into five subscales: pain, other symptoms, activities of daily living (ADL), sports and recreational function (Sport / Rec), and knee-related quality of life (QoL). Each subscale is scored separately on a scale of 0 to 100, with higher numbers indicating better status (Collins NJ, Misra D, Felson DT, et al (2011) Measures of knee function: International Knee Documentation Committee (IKDC) Subjective Knee Evaluation Form, Knee Injury and Osteoarthritis Outcome Score (KOOS), Knee Injury and Osteoarthritis Outcome Score Physical Function Short Form (KOOS-PS), Knee Outcome Survey Activities of Daily Living Scale (KOS-ADL), Lysholm Knee Scoring Scale, Oxford Knee Score (OKS), Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC), Activity Rating Scale (ARS), and Tegner Activity Score (TAS). Arthritis Care Res (Hoboken). p. S208-28).The PRO KOOS score is an expanded version of the WOMAC score traditionally used in OA clinical trials (KOOS User Guide 2003). The KOOS includes the WOMAC OA index LK3.0 in its full original form, and the WOMAC score is calculable. Thus, the KOOS score provides a more comprehensive assessment, since it also includes sports and recreational function, and knee-related quality of life. The KOOS requires approximately 10 minutes for participants to complete.

[0108] Numeric Rating Scale (NRS): Numeric Rating Scales for Pain (NRS), traditionally used to assess pain in clinical trials with a 24-hour recall period (Hawker GA, Mian S, Kendzerska T, et al(2011)Measures of adult pain:Visual Analog Scale for Pain(VAS Pain),Numeric Rating Scale for Pain(NRS Pain),McGill Pain Questionnaire(MPQ),Short-Form McGill Pain Questionnaire(SF-MPQ),Chronic Pain Grade Scale(CPGS),Short Form-36 Bodily Pain Scale(SF-36 BPS),and Measure of Intermittent and Constant Osteoarthritis Pain(ICOAP).Arthritis Care Res(Hoboken).p.S240-52), will be assessed at regular intervals to confirm eligibility and assess pain status throughout the study.

[0109] Widespread Pain Index (WPI): The Widespread Pain Index (WPI) will be assessed at screening only to exclude participants with substantial pain originating in areas other than the target knee, fibromyalgia, or other undiagnosed conditions that may interfere with pain assessment.

[0110] Pain diary: In addition, a pain diary will be completed by the participant starting at screening and each day during the 12-week study and will be sent to the CRF at each visit. This diary will be used to record baseline pain medications, rescue medications, and pain levels daily. Participants should rate their pain intensity at approximately the same time each day, at their choosing. NRS pain ratings in the diary and NRS pain ratings performed during study visits should be documented separately on the CRF. At each study visit from screening through day 84, participants should be provided with a new pain diary covering at least the time until their next scheduled visit. Prescription or use of pain medications should also be documented as concomitant medications per section 6.2.1.

[0111] Knee MRI: MRI will be obtained from the target knee to select participants with active synovitis and visualize cartilage and other structures through the knee. An imaging protocol will be developed to quantify changes in synovitis, exudate volume, and cartilage volume and thickness within the index region (i.e., the region where the most cartilage damage occurs in OA participants with KL2-3) during treatment. The index region is defined as the union of the femoral medial anterior (FMA), central (FMC), and posterior (FMP) cartilage subregions in the knee. This technique will demonstrate the efficacy of Compound IA in reducing knee inflammation and whether this response correlates with reduced pain. It can also be used to quantify changes in cartilage volume and thickness within the index region. Additionally, assessment of synovitis activity levels using dynamic contrast enhanced (DCE) MRI technique will demonstrate the efficacy of Compound IA in reducing knee inflammation and whether this response correlates with reduced pain.

[0112] Safety during the treatment period is supported by a 13-week GLP toxicology study in rats and cynomolgus monkeys. The expected mean steady-state daily systemic drug exposure in participants administered 25 mg bid with food will remain approximately 7-fold lower than the mean NOAEL plasma AUC in rats with an even larger safety margin for free AUC (14-fold) or Cmax (17-fold (total) and 34-fold (free)). Furthermore, the expected mean steady-state drug exposure (total and unbound) will remain at least 49-fold lower than the exposure recorded at the NOAEL dose (highest tested dose) of 150 mg / kg / day in monkeys.

[0113] Safety margin for Compound IA administered at 25 mg bid based on a 13-week GLP toxicology study:

[0114] [Table 4]

[0115] Drug-drug interaction challenges: Evaluation and recommendation for cytochrome P450 (CYP) substrates / modulators and drug-drug interaction clinical trials of Compound IA are based on in vitro / preclinical data and physiologically-based PK simulations. Compound IA is expected to be primarily eliminated via hepatic CYP-mediated metabolism with CYP2C9 (68%) and CYP3A4 (29%) as the major contributing enzymes. Participants who are poor CYP2C9 metabolizers will be excluded from the study.

[0116] Given the duration of treatment and the ample safety margin, administration of Compound IA is considered safe even under conditions of increased exposure to Compound IA.

[0117] Prohibited drugs and herbal remedies: - Anti-rejection / immunomodulatory therapy (e.g., anakinra, canakinumab or other investigational IL-1 / NLRP3 binding or blocking therapies) -Live vaccines -Strong or moderate inducers of CYP2C9 or strong inducers of CYP3A, including carbamazepine, enzalutamide, lumacaftor, phenobarbital, phenytoin, rifabutin, mitotane and St. John's wort (Hypericum perforatum). -Potent inhibitors of CYP2C9, including miconazole, berberine (botanical), sulfaphenazole, fluconazole, and resveratrol (botanical). - Other investigational drugs

[0118] Drugs to use with caution: -Drugs metabolized by CYP3A: In vitro metabolism studies have shown that Compound IA may have the potential to induce metabolism of drug substrates metabolized by the isoenzyme CYP3A. Thus, investigators may administer concomitant medications known to be metabolized by CYP3A4 / 5 at their discretion. Patients receiving such medications may require dose titration or increase in concomitant medication. Caution is advised, especially when Compound IA is coadministered with drugs that are sensitive substrates of CYP3A and / or have a narrow therapeutic index. -Medications that are strong or moderate inhibitors of CYP3A: Because Compound IA has been identified in vitro as a substrate of CYP3A, an increased systemic exposure of Compound IA cannot be excluded when coadministered with strong CYP3A inhibitors such as antivirals (e.g., ritonavir), antifungals (e.g., itraconazole, ketoconazole) and antibiotics (e.g., erythromycin, clarithromycin). Investigators may, at their discretion, coadminister known inhibitors of CYP3A, but their duration should be kept as short as possible and patients must be closely monitored.

[0119] Example 2: Clinical First-in-Human (FIH) Study: Test Design The study design consisted of four parts: single ascending dose (SAD; Part A), relative bioavailability of tablet formulations (Part B), multiple ascending dose (MAD; Part C), and relative bioavailability and food effect (Part D) (Figure 2; RF = reference formulation (crystal suspension); T2 = test formulation 2 (crystal tablet); T3 = test formulation 3 (spray-dried dispersion suspension); T4 = test formulation 4 (encapsulated crystal tablet)). In each group of Part A and Part C, eight subjects were randomized in a 3:1 ratio to receive Compound IA (6 subjects) or matching placebo (2 subjects).

[0120] For Part A, eight cohorts of eight eligible subjects were enrolled. Each subject received a single oral dose of Compound IA (3, 10, 30, 100, 300 mg of crystal suspension and 100, 300, 600 mg of spray-dried dispersion (SDD) under fasting conditions. As this was an FIH study, two sentinel subjects were dosed first, at least 24 hours prior to dosing the remainder of the cohort to ensure maximum safety. Part B was skipped as data from Part A provided an adequate comparison of the crystal and SDD formulations.

[0121] For Part C, eligible subjects were enrolled in six different cohorts. Each subject received multiple doses of Compound IA once daily (QD) under fasted conditions (10, 30 mg crystal suspension and 100, 200 mg SDD for 14 days) and under fed conditions (25, 50 mg encapsulated crystal tablet for 13 days and a single dose or placebo on day 14). Subjects were dosed in Part C following review of available safety, tolerability and PK data from the preceding arms in Part A.

[0122] Part D had an open-label, randomized, three-period crossover design with one group of six subjects. The PK of a crystalline tablet formulation of Compound IA was compared between fed and fasted conditions and with the PK of a crystalline suspension of Compound IA under fasted conditions. Subjects received three doses of Compound IA with a 7-14 day washout period 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 fasted conditions). Based on these doses, subjects were randomly assigned to one of six treatment sequences (one subject per sequence), prepared using a Williams design.

[0123] subject Eligible subjects were those with a body mass index (BMI) of ≥ 18.5 and ≤ 30.0 kg / m 2 The subjects were healthy men and women, aged 18-64 years. No subject participated in more than one 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 an overall high-fat breakfast during the specified time frame. Subjects were excluded if they had a history of major psychiatric disorder, a diagnosis of intellectual disability, clinically significant vital sign abnormalities, and tobacco product use within 90 days prior to (first) drug administration through follow-up.

[0124] Blinding In Parts A and C, the active and placebo treatments, which cannot be distinguished based on the label, were identical in appearance and had similar taste and odor. To maintain blinding, the same number of tablets or suspensions were administered to each subject in each cohort. Investigators and subjects remained blinded throughout the relevant part of the study, and blinding remained uninterrupted throughout. The sponsor (IFM Management, Inc.) was unblinded with access to all study data and was provided with a copy of the randomization code to support the decisions made regarding the study. Part D was unblinded, and only Compound IA was administered to subjects in one of six treatment sequences (one subject per sequence) according to the Williams design.

[0125] the purpose The primary objective of the study was to evaluate the safety and tolerability of oral administration of Compound IA at SAD and MAD in healthy subjects in all parts of the study. Key secondary objectives were to characterize the PK properties of Compound IA following single and multiple doses and to evaluate the effect of food on the PK properties of Compound IA.

[0126] evaluation Safety assessments in all parts of the study included adverse events (AEs) reported using the Medical Dictionary for Regulatory Activities (version 22.1), clinical laboratories (biochemistry, hematology, and urinalysis), vital signs, electrocardiogram (ECG), physical examination, and skin biopsy (when applicable).

[0127] In the single-dose part, blood samples were collected to measure Compound IA concentrations at the following time points for dosing on Day 1: pre-dose and 0.25, 0.5, 0.75, 1, 1.5, 2, 3, 4, 6, 8, 12, 24, 36, and 48 hours post-dose, and at the follow-up visit. In the multiple-dose part, samples were collected at pre-dose and 0.25, 0.5, 0.75, 1, 1.5, 2, 3, 4, 6, 8, and 12 hours post-dose for dosing on Days 1 and 14; for Days 2, 4, 7, 9, and 11: pre-dose; after the last dose on Day 14: 24 and 36 hours (Day 15), and 48 hours (Day 16); and at the follow-up visit. The following PK parameters were measured: maximum noncompartmental concentration in plasma (C max ); time to maximum concentration (t max ); Concentration 24 hours after administration (C 24h ) (Part A only); Lag time: observed time to first quantifiable concentration (t lag ); time to final quantifiable concentration (t last ); area under the concentration-time curve (AUC 0-last ); Area under the plasma concentration-time curve from 0 hours to infinity (AUC 0-inf); Area under the plasma concentration-time curve (AUC 0-24 ); terminal phase rate constant (K el ); Terminal half-life (t 1 / 2 ); apparent total body clearance (CL / F); and apparent volume of distribution in the terminal phase (V z / F); and additionally for Part C only: Area under the plasma concentration-time curve (AUC 0-tau ); Apparent clearance at steady state (CLss / F); AUC 0-tau Accumulation ratio based on (R ac , AUC); and C max Accumulation ratio based on (R ac , C max ) was used for evaluation.

[0128] To determine the PD response to NLRP3 inhibition, whole blood samples were collected and exploratory PD analyses were performed (Part A and Part C, groups 1–3). Ex vivo stimulation of the NLRP3 inflammasome by activating it with lipopolysaccharide (LPS) was assessed compared to control conditions, followed by analysis of blood cell release of the inflammatory marker IL-1β.

[0129] statistical analysis All data were summarized using descriptive statistics and listed and summarized in tabular and / or graphical format. Descriptive statistics for all relevant PK parameters included n, mean, standard deviation (SD), coefficient of variation (CV%), minimum, median, maximum, geometric mean, and geometric CV%. max In cases where only median, minimum and maximum values ​​are presented. PK parameters were calculated using non-compartmental methods using the software Phoenix Version 8.1. Concentrations below the lower limit of quantification (LLOQ) were treated as 0 in summary statistics limited to concentration data. For calculation of AUC, the linear trapezoidal method was used. t 1 / 2 Regression analysis of the terminal plasma removal phase to determine C max At least three data points were included after. Adjusted r 2 Parameters where %AUC was less than 0.80 were flagged but included in the descriptive statistics.extra AUC when > 20% 0-inf , %AUC extra , CL / F, and VZ / F parameters were excluded from the descriptive statistics.

[0130] In Part A, dose proportionality was evaluated using log-transformed C versus log-transformed dose levels. max , AUC 0-last , and AUC 0-inf The effect of food on the PK data was explored using an exponential regression model. Point estimates for the intercept and slope and corresponding 90% confidence intervals (CI) for the slope were calculated. Dose proportionality was not explored for Part C. In Part D, the relative bioavailability of the test formulations (crystal tablet, SDD, and crystal tablet) versus the reference formulation (crystal suspension), and the effect of food were explored using an analysis of variance (ANOVA) model for the PK data.

[0131] Least squares geometric mean ratios are presented with 90% CIs for the following treatments: fasted 100 mg Compound IA tablet over fasted 100 mg Compound IA suspension, fed 100 mg Compound IA tablet over fasted 100 mg Compound IA tablet.

[0132] Combined individual and mean plots of individual IL-1β concentrations versus time are presented for each treatment. Modeling the effect of Compound IA on corrected and stimulated ex vivo lipopolysaccharide (LPS) challenges in whole blood included evaluation of the relationship between LPS challenge outcomes by conditional weighted residual modeling.

[0133] result Subject characteristics and demographics A total of 122 subjects were enrolled in the study. All 122 subjects were included in the safety and PD analysis set, and all 94 subjects receiving active treatment (Compound IA) were included in the PK analysis set. Overall, subjects were aged 18-64 years and had a mean body weight of 18.9-29.4 kg / m 2Fifty-eight (48%) male and 64 (52%) female subjects with a BMI of 0.01 were enrolled in the study. The majority of subjects, 105 (86%) (Part A, n=57; Part C, n=42, and Part D, n=6), were white.

[0134] Of the enrolled subjects, 107 (88%) subjects completed the study according to protocol, and 15 (12%) subjects discontinued the study early. These early discontinuations included 1 of 64 (2%) subjects in Part A of the study, 13 of 52 (25%) subjects in Part C, and 1 of 6 (17%) subjects in Part D. Reasons for discontinuation included discontinuation due to adverse effects (AEs) in 12 (10%) subjects, and 1 (1%) subject each discontinued due to either withdrawal of consent, loss to follow-up, or temporary suspension of the study due to the COVID-19 pandemic (prevention of visits; unrelated to the safety of Compound IA). All 4 discontinued subjects were substituted into Part C.

[0135] safety Single and multiple doses of Compound IA were generally well tolerated. No deaths or serious adverse events (SAEs) were reported during the study. Overall, 87 / 122 subjects (71%) reported treatment-emergent adverse events (TEAEs); 66 / 94 subjects (70%) in the Compound IA arm, and 21 / 28 subjects (75%) in the placebo arm. The majority of TEAEs reported by 84 (69%) subjects were mild in intensity, while 15 subjects (12%) reported moderate TEAEs. Frequently reported system organ class AEs in >20% of subjects were nervous system disorders (34%), general disorders and administration site conditions (29%), and gastrointestinal disorders (27%).

[0136] Collectively, 46 relevant TEAEs reported by 24 / 122 subjects (20%) were considered related to study drug, including 21 / 94 (22%) subjects who received Compound IA and 3 / 28 (11%) subjects who received placebo. In the case of 12 / 122 (10%) subjects, 20 TEAEs of maculopapular rash and / or pruritus were considered to be adverse events of particular interest. All 12 subjects received Compound IA either as a single dose (100 mg [n=1] or 600 mg [n=1]) or multiple doses (30 mg QD [n=2], 100 mg QD [n=3], 200 mg QD [n=2], or 50 mg BID [n=3]). These TEAEs were mild to moderate in intensity, generally began within 1 to 17 days after initiating treatment with Compound IA, and resolved within 1 to 18 days after onset; all cases did not involve concomitant therapy. In 10 subjects, these TEAEs led to treatment discontinuation. The other two subjects discontinued early due to TEAEs unrelated to study drug.

[0137] Moderate decreases in neutrophil and white blood cell counts were considered non-clinically significant and were occasionally noted, which may be consistent with a PD effect of Compound IA due to inhibition of IL-1β signaling downstream of NLRP3. One subject had second-degree atrioventricular block that was not considered related to study drug. No other clinically significant findings were reported on vital signs, 12-lead ECG, 24-hour Holter monitoring, or physical examination.

[0138] Pharmacokinetics Exposure to a single dose of Compound IA increased in a less than dose-proportional manner when Compound IA was administered as a crystal suspension (3-300 mg), but increased dose-proportionally when Compound IA was administered as an SDD suspension (100-600 mg). After 2 weeks of QD administration of Compound IA in the dose range of 30-200 mg, only limited drug accumulation of approximately 1.1-1.3-fold was observed in reaching steady state. This was consistent with mean t 1 / 2At steady state, Compound IA showed very low CLss / F (~0.83-1.11 L / hr) and Vss / F (~12.6-23.3 L) with low to moderate intersubject variability across the QD and BID dose levels of Compound IA. Renal clearance at steady state was relatively low (~0.008 L / hr) compared to total oral clearance and therefore unlikely to be a significant clearance pathway in humans.

[0139] Administration of a single dose of 100 mg of Compound IA as a crystalline suspension under fed conditions significantly increased the C-terminal endoscopy rate of Compound IA compared to fasted conditions. max 2.05 times and AUC 0-last In the case of the crystalline tablets (100 mg of Compound IA under fasting conditions), the median t max is delayed for 2 to 5 hours, C max t 1 / 2 The encapsulated crystal tablet (25 mg and 50 mg bid under fed conditions) had a median lag time of 0.75 and 0.25 hours, respectively, and a median t of 4 hours on Day 1. max Compound IA was characterized by the average t 1 / 2 was comparable between the tablet (18.6 hours) and suspension (17.7 hours) formulations.

[0140] Pharmacodynamics A dose-dependent decrease in IL-1β concentrations (with mean nadir concentrations of approximately 5% to 20% of baseline values) was observed with increasing single and multiple oral doses of Compound IA. At the highest dose level of Compound IA, inhibition of IL-1β was observed from 1 hour post-dose to the final sampling time point for single (Day 3 or up to 6 hours for the lowest dose level ≦10 mg) and multiple (Day 15) oral doses of Compound IA.

[0141] The maximum stimulatory effect of the fractions, tested using the Hill coefficient (E max Based on the IL-1β model, the observed mean (±SD) stimulatory effect of IL-1β was 1820 (±102) ng / L, and the Emax The median potency (IC) of compound IA to inhibit 90% of in vitro stimulated IL-1β release upon lipopolysaccharide (LPS) challenge was −0.985 (±0.00277). 90 The effective concentration for the estimated maximum therapeutic effect and the inhibitory concentration for 100% inhibition of compound IA due to in vitro stimulated IL-1β release were EC 50 :0.141μM(90%CI:0.114,0.171), EC 90 :2.57μM(90%CI:2.24,2.94), and IC 50 :0.146μM (90%CI:0.118,0.179).

[0142] Consideration Single and multiple doses of Compound IA or placebo were generally well tolerated. No deaths or serious adverse events (SAEs) were reported during the study. TEAEs such as skin rash and / or pruritus were considered related to study drug. The majority of TEAEs reported by subjects were mild (69%) and moderate (12%) in severity. Maculopapular and / or pruritic skin rash were most frequently reported with Compound IA at the highest multiple dose levels, suggesting a relationship to exposure to Compound IA.

[0143] Following a single oral dose of Compound IA under fasting conditions, Compound IA demonstrated median t max However, at higher doses in the range of 30 to 600 mg, the median t max was somewhat delayed (1.5-3.0 hours), indicating slower absorption compared to lower doses (3 mg and 10 mg: 0.76 hours and 1.00 hours, respectively). The increase in drug exposure was less than dose-proportional for the crystalline suspensions (especially 100 and 300 mg), whereas there was a dose-proportional increase in exposure for the SDD suspensions (100-600 mg), indicating solubility-limited absorption of the crystalline material at doses ≥ 100 mg.

[0144] Multiple doses and formulations of Compound IA demonstrated no deviation from dose-proportional drug exposure after 2 weeks, indicating that the multiple-dose PK was linear and not solubility limited. Following oral administration of Compound IA on Day 1, a slight delay in absorption was observed for the encapsulated crystalline tablet under fed conditions. This slower absorption was observed at t max This was consistent with the bioavailability results where no significant food effect on absorption was observed. These findings suggest that the delayed absorption time was due to encapsulation. Renal clearance was determined to be approximately 0.004 L / hr (day 1) or 0.008 L / hr (day 14), approximating less than 0.8% of the oral dose. This indicates that direct excretion of the parent drug into the urine is not expected to be the primary elimination route for this drug in humans.

[0145] Compound IA as a 100 mg crystalline tablet is max The median T and AUC in the 100 mg crystalline tablet were increased by 2.05-fold and 1.49-fold, respectively, in the fed state (high-calorie, high-fat meal) versus the fasted state, indicating a positive food effect. max was 5 hours, whereas the suspension had a shorter T max Values ​​(0.76-3.0 hours) have been reported. Compound IA has a very low oral clearance (CLss / F of approximately 1.0 L / hour) associated with ≦2% of human hepatic blood flow, and a low volume of distribution (Vss / F) of approximately 12.6-23.3 L. When assessed for crystalline tablets at feeding time, slight drug accumulation of approximately 1.2-fold was observed after once daily dosing and 2-fold was observed after twice daily dosing in the process of reaching steady state, consistent with an effective half-life of approximately 10 hours.

[0146] Nonclinical studies suggest that compound IA blocks the release of IL-1β using a broad range of NLRP3-dependent activators. This has been observed in the case of diarylsulfonylurea compounds structurally similar to compound IA

[15] . In this study, a dose-dependent decrease in the concentration of IL-1β was observed with increasing single and multiple oral doses of compound IA. Although IL-1β production can be mediated by other inflammasomes or inflammasome-independent pathways; therefore, inhibitors directed at IL-1β may result in unintended immunosuppressive effects. Therefore, a pharmacological inhibitor that specifically targets exclusively the NLRP3 inflammasome may be a better option for the treatment of NLRP3-related diseases. Safety laboratory findings were mild, nonclinically significant reductions in neutrophil and white blood cell counts in 27 subjects. This may be consistent with the PD effect of compound IA resulting from inhibition of NLRP3 downstream signaling, similar to the known effect of the anti-IL-1β monoclonal antibody canakinumab.

[0147] The AUC in subjects with heterogeneous CYP2C9 genotypes was higher than that observed in subjects with normal CYP2C9 activity. These results suggest that the clearance of compound IA is influenced by the decreased CYP2C9 activity caused by certain genetic variants.

[0148] 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 concerns. The PK profile of Compound IA is compatible with a twice-daily dosing regimen. 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 that warrants further clinical evaluation.

[0149] Example 3: The following procedures are suitable for testing the activity of NLRP3 inhibitors according to the present disclosure.

[0150] Step 1: IL-1β production in PMA-differentiated THP-1 cells stimulated with gramicidin THP-1 cells were purchased from the American Type Culture Collection and passaged according to the supplier's instructions. Cells were cultured in complete RPMI 1640 (containing 10% heat-inactivated FBS, penicillin (100 units / ml) and streptomycin (100 μg / ml)) and maintained in log phase prior to experimental setup. Compounds were dissolved in dimethyl sulfoxide (DMSO) to make 30 mM stocks prior to the experiment. Compound stocks were first pre-diluted in DMSO to intermediate concentrations of 3, 0.34, 0.042 and 0.0083 mM and then spotted into empty 384-well assay plates using an Echo550 liquid handler to achieve the desired final concentrations (e.g., 100, 33, 11, 3.7, 1.2, 0.41, 0.14, 0.046, 0.015, 0.0051, 0.0017 μM). DMSO was back-filled into the plates to achieve a final DMSO assay concentration of 0.37%. Plates were then sealed and stored at room temperature until needed.

[0151] THP-1 cells were treated with PMA (phorbol 12-myristate 13-acetate) (20 ng / ml) for 16-18 hours. On the day of the experiment, the medium was removed and the adherent cells were detached with trypsin for 5 minutes. The cells were then collected, washed with complete RPMI 1640, spun down and resuspended in RPMI 1640 (containing 2% heat-inactivated FBS, penicillin (100 units / ml) and streptomycin (100 μg / ml)). The cells were plated at a density of 50,000 cells / well in 384-well assay plates containing spotted compounds (final assay volume 50 μl). The cells were incubated with compounds for 1 hour and then stimulated with gramicidin (5 μM) (Enzo) for 2 hours. The plates were then centrifuged at 340 g for 5 minutes. Cell-free supernatants (40 μL) were collected using a 96-channel PlateMaster (Gilson) and IL-1β production was assessed by HTRF (cisbio). Plates were incubated for 18 h at 4 °C and read using a preset HTRF program (donor emission at 620 nm, acceptor emission at 668 nm) on a SpectraMax i3x spectrophotometer (Molecular Devices, software SoftMax6). Vehicle-only controls and a dose titration of CRID3 (100-0.0017 μM) were run simultaneously with each experiment. Data were normalized to vehicle-treated samples (equal to 0% inhibition) and 100 μM CRID3 (equal to 100% inhibition). Compounds showed concentration-dependent inhibition of IL-1β production in PMA-differentiated THP-1 cells.

[0152] Step 2: IL-1β production in PMA-differentiated THP-1 cells stimulated with gramicidin THP-1 cells were purchased from the American Type Culture Collection and passaged according to the supplier's instructions. Cells were cultured in complete RPMI 1640 (containing 10% heat-inactivated FBS, penicillin (100 units / ml) and streptomycin (100 μg / ml)) and maintained in log phase before experimental setup. Prior to the experiment, THP-1 were treated with PMA (phorbol 12-myristate 13-acetate) (20 ng / ml) for 16-18 hours. Compounds were dissolved in dimethyl sulfoxide (DMSO) to make 30 mM stocks. On the day of the experiment, the medium was removed and adherent cells were detached with trypsin for 5 minutes. Cells were then harvested, washed with complete RPMI 1640, spun down, and resuspended in RPMI 1640 containing 2% heat-inactivated FBS, penicillin (100 units / ml), and streptomycin (100 μg / ml). Cells were plated at a density of 50,000 cells / well in 384-well plates (final assay volume 50 μl). Compounds were first dissolved in assay medium to obtain a 5× top concentration of 500 μM. Ten serial dilutions (1:3) were then performed in assay medium containing 1.67% DMSO. 5× compound solutions were added to medium to achieve the desired final concentrations (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 was 0.37%. Cells were incubated with compounds for 1 h and then stimulated with gramicidin (5 μM) (Enzo) for 2 h. Plates were then centrifuged at 340 g for 5 min. Cell-free supernatants (40 μL) were collected using a 96-channel PlateMaster (Gilson) and IL-1β production was assessed by HTRF (cisbio). Vehicle-only controls and dose titrations of CRID3 (100-0.0017 μM) were run simultaneously with each experiment. Data were normalized to vehicle-treated samples (equivalent to 0% inhibition) and 100 μM CRID3 (equivalent to 100% inhibition). Compounds showed concentration-dependent inhibition of IL-1β production in PMA-differentiated THP-1 cells.

[0153] Step 3: 1. Experimental Procedure: 1.1 Cell culture 1) Culture THP-1 cells in complete RPMI-1640 medium containing 10% FBS at 37 °C and 5% CO2. 2) 3 x 10 per ml 5 Passage the cells every 3 days by inoculating 100 cells. 1.2 Compound preparation Using the TECAN EVO system, prepare 3-fold serial dilutions of compounds in DMSO in 384-well LDV microplates to create the compound source plate using 10 concentrations. The highest concentration is 30 mM. 1.3 Cell preparation 1) Centrifuge THP-1 cells at 350g for 5 minutes. 2) Resuspend the cells in complete RMPI-1640 medium and count the cells. 3) Seed the cells into a T225 flask (approximately 2.5 × 10 7 cells / flask), and treat the cells with 20 ng / ml PMA (final DMSO concentration <1%). 4) Incubate overnight 1.4 THP-1 stimulation 1) Wash adherent THP-1 cells with PBS and detach the cells with 4 ml of trypsin in a T225 flask. 2) Centrifuge the cells at 350g for 5 minutes, resuspend the cells in RMPI-1640 medium containing 2% FBS, and count the cells with trypan blue. 3) Transfer 50nl / well of serial dilutions of test compounds to a 384 well plate by Echo; for advanced control and point 1 of CRID3 (MCC950), mirror 165nl and then backfill to make the DMSO concentration consistent in all wells, the plate layout is as follows: 4) Seed 50k cells in 40ul of RPMI-1640 with 2% FBS / well in a 384-well plate. 5) Incubate at 37℃, 5% CO2 for 1 hour. 6) Prepare 5x gramicidin, add 10 μl / well (final concentration is 5 μM) and incubate at 37°C, 5% CO2 for 2 hours. 7) Centrifuge at 350g for 1 minute. Pipette 16 μl of the supernatant using an apricot and transfer to a white 384 proxiplate. HC: 100 μM CRID3 (MCC950) + 5 μM gramicidin LC: 5 μM gramicidin 1.5 Detection of IL-1β 1) Homogenize 5x Diluent #5 by vortexing and add 1 volume of the stock solution to 4 volumes of distilled water. 2) Thaw 20x stocks of anti-IL1β-cryptate and anti-IL1β XL antibodies. Dilute these two antibodies to 1x in Detection Buffer #3. 3) Premix two ready-made antibody solutions immediately before use 4) Dispense 4 ul of premixed anti-IL1β antibody working solution into all wells. 5) Seal the plate and incubate at 4°C overnight. 6) The cell plate is read using EnVison and the readings are plotted versus test compound concentration to determine the IC 50 Calculate

[0154] 2. Data Analysis: 1. Compound IC 50 can be calculated using the following formula: I C 50 Formula %inhibition=100-100×[HC ave -Read / (HC ave -LC ave )] 2. Fit the normalized data in a dose-response fashion using XLfit to calculate compound concentrations

[0155] The table below shows the biological activity of compounds in the hTHP-1 assay with 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 = "+"

[0156] [Table 5]

[0157] All publications and patent documents cited in this specification are incorporated herein by reference as if each such publication and document was specifically and individually indicated to be incorporated herein by reference. The present invention and its embodiments have been described in detail. However, it is not intended that the scope of the present invention be limited to the particular embodiments of any process, manufacture, composition, compound, means, methods, and / or steps described herein. Various modifications, substitutions, and variations can be made to the disclosed material without departing from the spirit and / or essential characteristics of the present invention. Thus, one skilled in the art will readily appreciate from the present invention that subsequent modifications, substitutions, and / or variations that perform substantially the same function or achieve substantially the same results as the embodiments described herein can be utilized in accordance with such related embodiments of the present invention. Therefore, it is intended that the following claims include, within their scope, modifications, substitutions, and variations to the processes, manufacture, compositions, compounds, means, methods, and / or steps disclosed herein. The claims should be read as limited to the described order or elements unless otherwise stated to that effect. It should be understood that various changes in form and detail can be made without departing from the scope of the appended claims.

Claims

1. The NLRP3 inhibitor is administered to a subject at a total daily dose of about 10 mg to about 100 mg in a single dose or divided doses, for use in the treatment of osteoarthritis, or for use of the NLRP3 inhibitor in the manufacture of a medicament for the treatment of osteoarthritis.

2. The NLRP3 inhibitor according to claim 1, which is administered to a subject at a total daily dose of about 20 mg to about 50 mg in a single dose or divided doses.

3. The NLRP3 inhibitor according to claim 1 or 2, which is administered to a subject at a total daily dose of about 20 mg in a single dose or divided doses.

4. The NLRP3 inhibitor according to claim 1, which is administered to a subject at a total daily dose of about 50 mg in a single dose or divided doses.

5. The NLRP3 inhibitor according to claim 1, which is administered to a subject twice a day at a dose of about 10 mg.

6. The NLRP3 inhibitor according to claim 1, which is administered to a subject twice a day at a dose of about 10 mg for about 14 consecutive days.

7. The NLRP3 inhibitor according to claim 1, which is administered to a subject twice a day at a dose of about 25 mg.

8. The NLRP3 inhibitor according to claim 1, which is administered to a human subject twice a day at a dose of about 25 mg for about 70 consecutive days.

9. The NLRP3 inhibitor according to claim 1, which is administered to a subject during or after consumption of food.

10. The NLRP3 inhibitor according to claim 1, wherein there is a time interval of about 10 to 14 hours between administrations of two subsequent doses of the NLRP3 inhibitor to the subject.

11. The NLRP3 inhibitor according to claim 1, wherein the osteoarthritis is knee osteoarthritis.

12. The NLRP3 inhibitor according to claim 1, wherein when the administration of the NLRP3 inhibitor is determined by the KOOS score based on the change from baseline, it reduces pain in the joint affected by the osteoarthritis.

13. Administration of the NLRP3 inhibitor reduces the inflammation level of the joint affected by osteoarthritis when determined by the change from baseline in the activity level of synovitis measured by dynamic contrast-enhanced (DCE)-MRI. The NLRP3 inhibitor according to claim 1. ｔｒａｎｓ from baseline in the activity level of synovitis measured by dynamic contrast-enhanced (DCE)-MRI. The NLRP3 inhibitor according to claim 1.

14. The NLRP3 inhibitor according to claim 1, wherein the level of serum high-sensitivity C-reactive protein is reduced by 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% when determined by the change from baseline in the subject.

15. The level of IL-1β or IL-18 is reduced by 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% when determined by the change from baseline in a subject, the NLRP3 inhibitor according to claim 1.

16. The NLRP3 inhibitor according to claim 1, wherein the subject shows no rash at all.

17. The NLRP3 inhibitor according to claim 1, which is orally administered to the subject.

18. The NLRP3 inhibitor according to claim 1, which is contained in a tablet formulation.

19. The NLRP3 inhibitor according to claim 1, comprising administering at least one additional therapeutic agent.

20. Compound I, or a pharmaceutically acceptable salt thereof: 【Chemical 1】 The NLRP3 inhibitor according to claim 1, which is.

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