Patent Application
20240174670
The present invention relates to novel triazinones that are useful as inhibitors of NOD-like receptor protein 3 (NLRP3) inflammasome pathway. The present invention also relates to processes for the preparation of said compounds. pharmaceutical compositions comprising said compounds, methods of using said compounds in the treatment of various diseases and disorders, and medicaments containing them, and their use in diseases and disorders mediated by NLRP3.
Inflammasomes, considered as central signalling hubs of the innate immune system, are multi-protein complexes that are assembled upon activation of a specific set of intracellular pattern recognition receptors (PRRs) by a wide variety of pathogen- or danger-associated molecular patterns (PAMPs or DAMPs). To date, it was shown that inflammasomes can be formed by nucleotide-binding oligomerization domain (NOD)-like receptors (NLRs) and Pyrin- and HIN200-domain-containing proteins (Van Opdenbosch N. and Lamkanfi M. Immunity, 2019 Jun. 18:50(6):1352-1364). The NLRP3 inflammasome is assembled upon detection of environmental crystals, pollutants, host-derived DAMPs and protein aggregates (Tartey S and Kanneganti TD. Immunology, 2019 April: 156(4):329-338). Clinically relevant DAMPs that engage NLRP3 include uric acid and cholesterol crystals that cause gout and atherosclerosis, amyloid-β fibrils that are neurotoxic in Alzheimer's disease and asbestos particles that cause mesothelioma
(Kelley et al., Int J Mol Sci. 2019 Jul. 6:20(13)). Additionally, NLRP3 is activated by infectious agents such as Vibrio cholerae: fungal pathogens such as Aspergillus fumigatus and Candida albicans: adenoviruses, influenza A virus and SARS-COV-2 (Tartey and Kanneganti, 2019 (see above): Fung et al. Emerg Microbes Infect, 2020 Mar. 14:9(1):558-570)).
Although the precise NLRP3 activation mechanism remains unclear, for human monocytes, it has been suggested that a one-step activation is sufficient while in mice a two-step mechanism is in place. Given the multitude in triggers, the NLRP3 inflammasome requires add-on regulation at both transcriptional and post-transcriptional level (Yang Y et al., Cell Death Dis, 2019 Feb. 12:10(2): 128).
The NLRP3 protein consists of an N-terminal pyrin domain, followed by a nucleotide-binding site domain (NBD) and a leucine-rich repeat (LRR) motif on C-terminal end (Sharif et al., Nature, 2019 June: 570(7761):338-343). Upon recognition of PAMP or DAMP, NLRP3 aggregates with the adaptor protein, apoptosis-associated speck-like protein (ASC), and with the protease caspase-1 to form a functional inflammasome. Upon activation, procaspase-1 undergoes autoproteolysis and consequently cleaves gasdermin D (Gsdmd) to produce the N-terminal Gsdmd molecule that will ultimately lead to pore-formation in the plasma membrane and a lytic form of cell death called pyroptosis. Alternatively, caspase-1 cleaves the pro-inflammatory cytokines pro-IL-Iβ and pro-IL-18 to allow release of its biological active form by pyroptosis (Kelley et al., 2019—see above).
Dysregulation of the NLRP3 inflammasome or its downstream mediators are associated with numerous pathologies ranging from immune/inflammatory diseases. auto-immune/auto-inflammatory diseases (Cryopyrin-associated Periodic Syndrome (Miyamae T. Paediatr Drugs. 2012 Apr 1:14(2): 109-17); sickle cell disease: systemic lupus erythematosus (SLE)) to hepatic disorders (e.g. non-alcoholic steatohepatitis (NASH), chronic liver disease, viral hepatitis, alcoholic steatohepatitis, and alcoholic liver disease) (Szabo G and Petrasek J. Nat Rev Gastroenterol Hepatol, 2015 July: 12(7):387-400) and inflammatory bowel diseases (e.g. Crohn's disease, ulcerative colitis) (Zhen Y and Zhang H. Front Immunol. 2019 Feb. 28:10:276). Also, inflammatory joint disorders (e.g. gout. pseudogout (chondrocalcinosis), arthropathy. osteoarthritis. and rheumatoid arthritis (Vande Walle L et al., Nature. 2014 Aug 7:512(7512):69-73) were linked to NLRP3. Additionally, kidney related diseases (hyperoxaluria (Knauf et al., Kidney Int. 2013 Nov:84(5):895-901), lupus nephritis, hypertensive nephropathy (Krishnan et al., Br J Pharmacol. 2016 Feb: 173(4): 752-65), haemodialysis related inflammation and diabetic nephropathy which is a kidney-related complication of diabetes (Type 1, Type 2 and mellitus diabetes). also called diabetic kidney disease (Shahzad et al., Kidney Int. 2015 January:87(1):74-84) are associated to NLRP3 inflammasome activation. Reports link onset and progression of neuroinflammation-related disorders (e.g. brain infection, acute injury, multiple sclerosis. Alzheimer's disease) and neurodegenerative diseases (Parkinson's disease) to NLRP3 inflammasome activation (Sarkar et al., NPJ Parkinson's Dis. 2017 Oct. 17:3:30). In addition. cardiovascular or metabolic disorders (e.g. cardiovascular risk reduction (CvRR). atherosclerosis, type I and type II diabetes and related complications (e.g. nephropathy. retinopathy), peripheral artery disease (PAD), acute heart failure and hypertension (Ridker et al., CANTOS Trial Group. N Engl J Med. 2017 Sep 21:377(12): 1119-1131: and Toldo S and Abbate A. Nat Rev Cardiol. 2018 Apr: 15(4): 203-214) have recently been associated to NLRP3. Also, skin associated diseases were described (e.g. wound healing and scar formation: inflammatory skin diseases. e.g. acne, hidradenitis suppurativa (Kelly et al., Br J Dermatol. 2015 Dec: 173(6)). In addition, respiratory conditions have been associated with NLRP3 inflammasome activity (e.g. asthma, sarcoidosis, Severe Acute Respiratory Syndrome (SARS) (Nieto-Torres et al., Virology. 2015 Nov:485:330-9)) but also age-related macular degeneration (Doyle et al., Nat Med, 2012 May:18(5): 791-8). Several cancer related diseases/disorders were described linked to NLRP3 (e.g. myeloproliferative neoplasms, leukemias, myelodysplastic syndromes (MOS), myelofibrosis, lung cancer, colon cancer (Ridker et al., Lancet, 2017 Oct. 21:390(10105): 1833-1842; Derangere et al., Cell Death Differ. 2014 December:21(12):1914-24; Basiorka et al., Lancet Haematol. 2018 Sep:5(9): e393-e402, Zhang et al., Hum Immunol, 2018 January:79(1):57-62).
Several patent applications describe NLRP3 inhibitors, with recent ones including for instance international patent application WO 2020/234715, WO 2020/018975, WO 2020/037116, WO 2020/021447, WO 2020/010143, WO 2019/079119, WO 2019/0166621 and WO 2019/121691, which disclose a range of specific compounds.
There is a need for inhibitors of the NLRP3 inflammasome pathway to provide new and/or alternative treatments for the diseases/disorders mentioned herein.
The invention provides compounds which inhibit the NLRP3 inflammasome pathway.
Thus, in an aspect of the invention, there is now provided a compound of formula (I),
or a pharmaceutically acceptable salt thereof, wherein:
R1 represents: (i) phenyl: (ii) a 6-membered mono-cyclic heteroaryl group: or (iii) a 9-or 10-membered bicyclic heteroaryl group, all of which are optionally substituted with one or two substituent(s) selected from halo, —OH, C1-3 alkyl and —OC1-3 alkyl:
R2 represents:
In another aspect of the invention, there is provided a compound of formula (I),
or a pharmaceutically acceptable salt thereof, wherein:
R1 represents: (i) phenyl; (ii) a 6-membered mono-cyclic heteroaryl group; or (iii) a 9-or 10-membered bicyclic heteroaryl group, all of which are optionally substituted with one or two substituent(s) selected from halo, —OH, C1-3 alkyl and —OC1-3 alkyl;
R2 represents:
In another aspect, there is provided compounds of the invention for use as a medicament. In another aspect, there is provided a pharmaceutical composition comprising a therapeutically effective amount of a compound of the invention.
In a further aspect, there is provided compounds of the invention (and/or pharmaceutical compositions comprising such compounds) for use in the treatment of a disease or disorder associated with NLRP3 activity (including inflammasome activity): in the treatment of a disease or disorder in which the NLRP3 signalling contributes to the pathology, and/or symptoms, and/or progression, of said disease/disorder; in inhibiting NLRP3 inflammasome activity (including in a subject in need thereof): and/or as an NLRP3 inhibitor. Specific diseases or disorders may be mentioned herein, and may for instance be selected from inflammasome-related diseases or disorders, immune diseases, inflammatory diseases, auto-immune diseases, or auto-inflammatory diseases.
In another aspect, there is provided a use of compounds of the invention (and/or pharmaceutical compositions comprising such compounds): in the treatment of a disease or disorder associated with NLRP3 activity (including inflammasome activity); in the treatment of a disease or disorder in which the NLRP3 signalling contributes to the pathology, and/or symptoms, and/or progression, of said disease/disorder: in inhibiting NLRP3 inflammasome activity (including in a subject in need thereof): and/or as an NLRP3 inhibitor.
In another aspect, there is provided use of compounds of the invention (and/or pharmaceutical compositions comprising such compounds) in the manufacture of a medicament for: the treatment of a disease or disorder associated with NLRP3 activity (including inflammasome activity); the treatment of a disease or disorder in which the NLRP3 signalling contributes to the pathology, and/or symptoms, and/or progression, of said disease/disorder; and/or inhibiting NLRP3 inflammasome activity (including in a subject in need thereof).
In another aspect, there is provided a method of treating a disease or disorder in which the NLRP3 signalling contributes to the pathology, and/or symptoms, and/or progression, of said disease/disorder, comprising administering a therapeutically effective amount of a compound of the invention, for instance to a subject (in need thereof). In a further aspect there is provided a method of inhibiting the NLRP3 inflammasome activity in a subject (in need thereof), the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of the invention.
In further aspect, there is a provided a compound of the invention in combination (including a pharmaceutical combination) with one or more therapeutic agents (for instance as described herein). Such combination may also be provided for use as described herein in respect of compounds of the invention, or, a use of such combination as described herein in respect of compounds of the invention. There may also be provided methods as described herein in respect of compounds of the invention, but wherein the method comprises administering a therapeutically effective amount of such combination.
The invention provides a compound of formula (I),
or a pharmaceutically acceptable salt thereof, wherein:
R1 represents:
In another aspect of the invention, there is provided a compound of formula (I),
or a pharmaceutically acceptable salt thereof, wherein:
R1 represents:
Pharmaceutically-acceptable salts include acid addition salts and base addition salts. Such salts may be formed by conventional means, for example by reaction of a free acid or a free base form of a compound of the invention with one or more equivalents of an appropriate acid or base, optionally in a solvent, or in a medium in which the salt is insoluble, followed by removal of said solvent, or said medium, using standard techniques (e.g. in vacuo, by freeze-drying or by filtration). Salts may also be prepared by exchanging a counter-ion of a compound of the invention in the form of a salt with another counter-ion, for example using a suitable ion exchange resin.
Pharmaceutically acceptable acid addition salts can be formed with inorganic acids and organic acids.
Inorganic acids from which salts can be derived include, for example. hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid. and the like.
Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, mandelic acid. methanesulfonic acid, ethanesulfonic acid, toluenesulfonic acid, sulfosalicylic acid, and the like.
Pharmaceutically acceptable base addition salts can be formed with inorganic and organic bases.
Inorganic bases from which salts can be derived include, for example. ammonium salts and metals from columns I to XII of the periodic table. In certain embodiments, the salts are derived from sodium, potassium, ammonium. calcium, magnesium, iron, silver, zinc, and copper: particularly suitable salts include ammonium, potassium, sodium, calcium and magnesium salts.
Organic bases from which salts can be derived include, for example. primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, basic ion exchange resins, and the like. Certain organic amines include isopropylamine, benzathine, cholinate, diethanolamine, diethylamine, lysine, meglumine, piperazine, and tromethamine
For the purposes of this invention solvates, prodrugs, N-oxides and stereoisomers of compounds of the invention are also included within the scope of the invention.
The term “prodrug” of a relevant compound of the invention includes any compound that, following oral or parenteral administration, is metabolised in vivo to form that compound in an experimentally-detectable amount, and within a predetermined time (e.g. within a dosing interval of between 6 and 24 hours (i.e. once to four times daily)). For the avoidance of doubt, the term “parenteral” administration includes all forms of administration other than oral administration.
Prodrugs of compounds of the invention may be prepared by modifying functional groups present on the compound in such a way that the modifications are cleaved, in vivo when such prodrug is administered to a mammalian subject. The modifications typically are achieved by synthesising the parent compound with a prodrug substituent. Prodrugs include compounds of the invention wherein a hydroxyl, amino, sulfhydryl, carboxy or carbonyl group in a compound of the invention is bonded to any group that may be cleaved in vivo to regenerate the free hydroxyl, amino, sulfhydryl, carboxy or carbonyl group, respectively.
Examples of prodrugs include, but are not limited to, esters and carbamates of hydroxy functional groups, esters groups of carboxyl functional groups, N-acyl derivatives and N-Mannich bases. General information on prodrugs may be found e.g. in Bundegaard, H. “Design of Prodrugs” p. 1-92, Elsevier, New York-Oxford (1985).
Compounds of the invention may contain double bonds and may thus exist as E (entgegen) and Z (zusammen) geometric isomers about each individual double bond. Positional isomers may also be embraced by the compounds of the invention. All such isomers (e.g. if a compound of the invention incorporates a double bond or a fused ring, the cis- and trans-forms, are embraced) and mixtures thereof are included within the scope of the invention (e.g. single positional isomers and mixtures of positional isomers may be included within the scope of the invention).
Compounds of the invention may also exhibit tautomerism. All tautomeric forms (or tautomers) and mixtures thereof are included within the scope of the invention. The term “tautomer” or “tautomeric form” refers to structural isomers of different energies which are interconvertible via a low energy barrier. For example, proton tautomers (also known as prototropic tautomers) include interconversions via migration of a proton, such as keto-enol and imine-enamine isomerisations. Valence tautomers include interconversions by reorganisation of some of the bonding electrons.
Compounds of the invention may also contain one or more asymmetric carbon atoms and may therefore exhibit optical and/or diastereoisomerism. Diastereoisomers may be separated using conventional techniques, e.g. chromatography or fractional crystallisation. The various stereoisomers may be isolated by separation of a racemic or other mixture of the compounds using conventional, e.g. fractional crystallisation or HPLC, techniques. Alternatively the desired optical isomers may be made by reaction of the appropriate optically active starting materials under conditions which will not cause racemisation or epimerisation (i.e. a ‘chiral pool’ method), by reaction of the appropriate starting material with a ‘chiral auxiliary’ which can subsequently be removed at a suitable stage, by derivatisation (i.e. a resolution, including a dynamic resolution), for example with a homochiral acid followed by separation of the diastereomeric derivatives by conventional means such as chromatography, or by reaction with an appropriate chiral reagent or chiral catalyst all under conditions known to the skilled person.
All stereoisomers (including but not limited to diastereoisomers, enantiomers and atropisomers) and mixtures thereof (e.g. racemic mixtures) are included within the scope of the invention.
In the structures shown herein, where the stereochemistry of any particular chiral atom is not specified, then all stereoisomers are contemplated and included as the compounds of the invention. Where stereochemistry is specified by a solid wedge or dashed line representing a particular configuration, then that stereoisomer is so specified and defined.
When an absolute configuration is specified, it is according to the Cahn-Ingold-Prelog system. The configuration at an asymmetric atom is specified by either R or S. Resolved compounds whose absolute configuration is not known can be designated by (+) or (−) depending on the direction in which they rotate plane polarized light.
When a specific stereoisomer is identified, this means that said stereoisomer is substantially free, i.e. associated with less than 50%, preferably less than 20%, more preferably less than 10%, even more preferably less than 5%, in particular less than 2% and most preferably less than 1%, of the other isomers. Thus, when a compound of formula (I) is for instance specified as (R), this means that the compound is substantially free of the (S) isomer.
The compounds of the present invention may exist in unsolvated as well as solvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like, and it is intended that the invention embrace both solvated and unsolvated forms.
The present invention also embraces isotopically-labelled compounds of the present invention which are identical to those recited herein, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature (or the most abundant one found in nature). All isotopes of any particular atom or element as specified herein are contemplated within the scope of the compounds of the invention. Exemplary isotopes that can be incorporated into compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine and iodine, such as 2H, 3H, 11C, 13C, 14C , 13N, 15O, 17O, 18O, 32P, 33P, 35S, 18F, 36Cl, 123I, and 125I. Certain isotopically-labelled compounds of the present invention (e.g., those labelled with 3H and 14C) are useful in compound and for substrate tissue distribution assays. Tritiated (3H) and carbon-14 (14C) isotopes are useful for their ease of preparation and detectability. Further, substitution with heavier isotopes such as deuterium (i.e., 2H may afford certain therapeutic advantages resulting from greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements) and hence may be preferred in some circumstances. Positron emitting isotopes such as 15O, 13N, 11C and 18F are useful for positron emission tomography (PET) studies to examine substrate receptor occupancy. Isotopically labelled compounds of the present invention can generally be prepared by following procedures analogous to those disclosed in the description/Examples hereinbelow, by substituting an isotopically labelled reagent for a non-isotopically labelled reagent.
Unless otherwise specified, C1-q alkyl groups (where q is the upper limit of the range) defined herein may be straight-chain or, when there is a sufficient number (i.e. a minimum of two or three, as appropriate) of carbon atoms, be branched-chain. Such a group is attached to the rest of the molecule by a single bond.
C2-q alkenyl when used herein (again where q is the upper limit of the range) refers to an alkyl group that contains unsaturation, i.e. at least one double bond.
C3-q cycloalkyl (where q is the upper limit of the range) refers to an alkyl group that is cyclic, for instance cycloalkyl groups may be monocyclic or, if there are sufficient atoms, bicyclic. In an embodiment, such cycloalkyl groups are monocyclic. Such cycloalkyl groups are unsaturated. Substituents may be attached at any point on the cycloalkyl group.
The term “halo”, when used herein, preferably includes fluoro, chloro, bromo and iodo.
C1-4 alkoxy groups (where q is the upper limit of the range) refers to the radical of formula -ORa, where Ra is a C1-4 alkyl group as defined herein.
HaloC1-q alkyl (where q is the upper limit of the range) groups refer to C1-q alkyl groups, as defined herein, where such group is substituted by one or more halo. HydroxyC1-q alkyl (where q is the upper limit of the range) refers to C1-q alkyl groups, as defined herein, where such group is substituted by one or more (e.g. one) hydroxy (—OH) groups (or one or more, e.g. one, of the hydrogen atoms is replaced with —OH). Similarly, haloC1-q alkoxy and hydroxyC1-q alkoxy represent corresponding —OC1-q alkyl groups that are substituted by one or more halo, or, substituted by one or more (e.g. one) hydroxy, respectively.
Heterocyclyl groups that may be mentioned include non-aromatic monocyclic and bicyclic heterocyclyl groups in which at least one (e.g. one to four) of the atoms in the ring system is other than carbon (i.e. a heteroatom), and in which the total number of atoms in the ring system is between 3 and 20 (e.g. between three and ten, e.g. between 3 and 8, such as 5- to 8-). Such heterocyclyl groups may also be bridged. Such heterocyclyl groups are saturated. C2-4 heterocyclyl groups that may be mentioned include 7-azabicyclo[2.2.1 ]heptanyl, 6-azabicyclo[3.1.1 ]heptanyl, 6-azabicyclo[3.2.1]-octanyl, 8-azabicyclo-[3.2.1 ]octanyl, aziridinyl, azetidinyl, dihydropyranyl, dihydropyridyl, dihydropyrrolyl (including 2,5-dihydropyrrolyl), dioxolanyl (including 1,3-dioxolanyl), dioxanyl (including 1,3-dioxanyl and 1,4-dioxanyl), dithianyl (including 1,4-dithianyl), dithiolanyl (including 1,3-dithiolanyl), imidazolidinyl, imidazolinyl, morpholinyl, 7-oxabicyclo[2.2.1]heptanyl, 6-oxabicyclo-[3.2.1]octanyl, oxetanyl, oxiranyl, piperazinyl, piperidinyl, non-aromatic pyranyl, pyrazolidinyl, pyrrolidinonyl, pyrrolidinyl, pyrrolinyl, quinuclidinyl, sulfolanyl, 3-sulfolenyl, tetrahydropyranyl, tetrahydrofuranyl, tetrahydropyridyl (such as 1,2,3,4-tetrahydropyridyl and 1,2,3,6-tetrahydropyridyl), thietanyl, thiiranyl, thiolanyl, thiomorpholinyl, trithianyl (including 1,3,5-trithianyl), tropanyl and the like. Substituents on heterocyclyl groups may, where appropriate, be located on any atom in the ring system including a heteroatom. The point of attachment of heterocyclyl groups may be via any atom in the ring system including (where appropriate) a heteroatom (such as a nitrogen atom), or an atom on any fused carbocyclic ring that may be present as part of the ring system. Heterocyclyl groups may also be in the N- or S-oxidised form. In an embodiment, heterocyclyl groups mentioned herein are monocyclic.
Aryl groups that may be mentioned include C6-20, such as C6-12 (e.g. C6-10) aryl groups. Such groups may be monocyclic, bicyclic or tricyclic and have between 6 and 12 (e.g. 6 and 10) ring carbon atoms, in which at least one ring is aromatic. C6-10 aryl groups include phenyl, naphthyl and the like, such as 1,2,3,4-tetrahydronaphthyl. The point of attachment of aryl groups may be via any atom of the ring system. For example. when the aryl group is polycyclic the point of attachment may be via atom including an atom of a non-aromatic ring. However, when aryl groups are polycyclic (e.g. bicyclic or tricyclic), they are preferably linked to the rest of the molecule via an aromatic ring. When aryl groups are polycyclic, in an embodiment, each ring is aromatic. In an embodiment, aryl groups mentioned herein are monocyclic or bicyclic. In a further embodiment, aryl groups mentioned herein are monocyclic.
“Heteroaryl” when used herein refers to an aromatic group containing one or more heteroatom(s) (e.g. one to four heteroatoms) preferably selected from N, O and S. Heteroaryl groups include those which have between 5 and 20 members (e.g. between 5 and 10) and may be monocyclic, bicyclic or tricyclic, provided that at least one of the rings is aromatic (so forming, for example, a mono-, bi-, or tricyclic heteroaromatic group). When the heteroaryl group is polycyclic the point of attachment may be via any atom including an atom of a non-aromatic ring. However, when heteroaryl groups are polycyclic (e.g. bicyclic or tricyclic), they are preferably linked to the rest of the molecule via an aromatic ring. In an embodiment, when heteroaryl groups are polycyclic, then each ring is aromatic. Heteroaryl groups that may be mentioned include 3,4-dihydro-1H-isoquinolinyl, 1.3-dihydroisoindolyl. 1,3-dihydroisoindolyl (e.g. 3,4-dihydro-1H-isoquinolin-2-yl, 1.3-dihydroisoindol-2-yl, 1,3-dihydroisoindol-2-yl: i.e. heteroaryl groups that are linked via a non-aromatic ring), or, preferably. acridinyl, benzimidazolyl, benzodioxanyl, benzodioxepinyl, benzodioxolyl (including 1,3-benzodioxolyl), benzofuranyl, benzofurazanyl, benzothiadiazolyl (including 2,1,3-benzothiadiazolyl), benzothiazolyl, benzoxadiazolyl (including 2,1,3-benzoxadiazolyl), benzoxazinyl (including 3,4-dihydro-2H-1,4-benzoxazinyl), benzoxazolyl, benzomorpholinyl, benzoselenadiazolyl (including 2,1,3-benzoselenadiazolyl), benzothienyl, carbazolyl, chromanyl, cinnolinyl, furanyl, imidazolyl, imidazo| 1,2-alpyridyl, indazolyl, indolinyl, indolyl, isobenzofuranyl, isochromanyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiaziolyl, isothiochromanyl, isoxazolyl, naphthyridinyl (including 1,6-naphthyridinyl or, preferably, 1,5-naphthyridinyl and 1,8-naphthyridinyl), oxadiazolyl (including 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl and 1,3,4-oxadiazolyl), oxazolyl, phenazinyl, phenothiazinyl, phthalazinyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolyl, pyridavinyl, pyridyl, pyrimidinyl, pyrrolyl, quinazolinyl, quinolinyl, quinolizinyl, quinoxalinyl, tetrahydroisoquinolinyl (including 1,2,3,4-tetrahydroisoquinolinyl and 5,6,7,8-tetrahydroisoquinolinyl), tetrahydroquinolinyl (including 1,2,3,4-tetrahydroquinolinyl and 5,6,7,8-tetrahydroquinolinyl), tetrazolyl, thiadiazolyl (including 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl and 1,3,4-thiadiazolyl), thiazolyl, thiochromanyl, thiophenetyl, thienyl, triazolyl (including 1,2,3-triazolyl, 1,2,4-triazolyl and 1,3,4-triazolyl) and the like. Substituents on heteroaryl groups may, where appropriate, be located on any atom in the ring system including a heteroatom. The point of attachment of heteroaryl groups may be via any atom in the ring system including (where appropriate) a heteroatom (such as a nitrogen atom), or an atom on any fused carbocyclic ring that may be present as part of the ring system. Heteroaryl groups may also be in the N- or S- oxidised form. When heteroaryl groups are polycyclic in which there is a non-aromatic ring present, then that non-aromatic ring may be substituted by one or more ═O group. In an embodiment, heteroaryl groups mentioned herein may be monocyclic or bicyclic. In a further embodiment, heteroaryl groups mentioned herein are monocyclic.
Heteroatoms that may be mentioned include phosphorus, silicon, boron and, preferably, oxygen, nitrogen and sulfur.
For the avoidance of doubt, where it is stated herein that a group may be substituted by one or more substituents (e.g. selected from C1-6 alkyl), then those substituents (e.g. alkyl groups) are independent of one another. That is, such groups may be substituted with the same substituent (e.g. same alkyl substituent) or different (e.g. alkyl) substituents.
All individual features (e.g. preferred features) mentioned herein may be taken in isolation or in combination with any other feature (including preferred feature) mentioned herein (hence, preferred features may be taken in conjunction with other preferred features, or independently of them).
The skilled person will appreciate that compounds of the invention that are the subject of this invention include those that are stable. That is, compounds of the invention include those that are sufficiently robust to survive isolation from e.g. a reaction mixture to a useful degree of purity.
Various embodiments of the invention will now be described, including embodiments of the compounds of the invention.
In an embodiment where R1 represents aryl or heteroaryl, optionally substituted as defined herein, then it may represent: (i) phenyl: (ii) a 5- or 6-membered mono-cyclic heteroaryl group: or (iii) a 9- or 10-membered bicyclic heteroaryl group, all of which are optionally substituted by one to three substituents as defined herein. In an embodiment, the aforementioned aryl and heteroaryl groups are optionally substituted with one or two (e.g. one) substituent(s) selected from halo (e.g. fluoro), —OH, C1-3 alkyl and —OC1-3 alkyl. In one embodiment, R1 represents phenyl or a mono-cyclic 6-membered heteroaryl group and in another embodiment it may represent a 9- or 10-membered (e.g. 9-membered) bicyclic heteroaryl group. Hence, in an embodiment, R1 may represent:
wherein R16 represents one or two optional substituents selected from halo, —CH3, —OH and —OCH3 (and in a further embodiment, such optional substituents are selected from fluoro and methoxy), and at least one of Rb, Rc, Rd, Re and Rf represents a nitrogen heteroatom (and the others represent CH). In an embodiment, either one or two of Rb, Rc, Rd, Re and Rf represent(s) a nitrogen heteroatom, for instance, Rd represents nitrogen and, optionally, Rb represents nitrogen, or, Rc represents nitrogen. In an aspect: (i) Rb and Rd represent nitrogen; (ii) Rd represents nitrogen; or (iii) Re represents nitrogen. Hence, R1 may represent 3-pyridyl or 4-pyrimidinyl, both of which are optionally substituted as herein defined; however, in an embodiment, such groups are unsubstituted.
In another embodiment, R1 may represent:
wherein R1b is as defined above (i.e. represents one or two optional substituents as defined above), each ring of the bicyclic system is aromatic, Rg represents a N or C atom and any one or two of Rh, Ri and Rj (for instance, one or two of Ri and Rj) represents N and the other(s) represent(s) C (provided that, as the skilled person would understand, the rules of valency are adhered to; for instance when one of the atoms of the (hetero)aromatic ring represents C, then it is understood that it may bear a H atom).
In an embodiment R1 represents:
in which Rb and Ra represent a nitrogen atom, and, in an embodiment, there is no R1b substituent present.
In another embodiment, R1 represents:
in which one of Ri and Rj represents N and the other represents C, or, both Ri and Rj represent N, and, in an embodiment, there is no R1b substituent present.
In an embodiment R2 represents: (i) C1-3 alkyl optionally substituted with one or more substituents independently selected from halo (e.g. fluoro), —OH and —OC1-2 alkyl; (ii) C3-6 cycloalkyl; or (iii) OC2-4 alkenyl optionally substituted by —OC1-2 alkyl. In a further embodiment, R2 represents C1-3 alkyl optionally substituted with one or more substituents independently selected from halo, —OH and —OC1-2 alkyl, or, R2 represents C3-6 cycloalkyl. In yet a further embodiment, R2 represents unsubstituted C1-3 alkyl or C3-6 cycloalkyl. In yet a further embodiment, R2 represents unsubstituted C1-3 alkyl.
In an embodiment, R3a and R3b are linked together to form a ring as hereinbefore defined, i.e. a 3- to 10-membered ring, which, in an embodiment, may be represented as follows:
where Sub represents one or more optional substituents that may be present on the cyclic group so formed (as hereinbefore defined); for instance Sub may represent one or more optional substituents selected from halo, C1-3 alkyl, —OH, —OC1-3 alkyl, —N(C1-3 alkyl)2, -C1-3alkyl—OH and -C1-3 alkyl—O-C1-3 alkyl: in a particular embodiment, Sub represents one or more optional substituents selected from halo (e.g. fluoro), —CH3, —CH2CH3, —CH3CH2, —OCH3, —OH, —N(CH3)2, —-CH2—OH and —CH2—O—CH3.
In an embodiment, one of R3a and R3b represents hydrogen or C1-3 alkyl (e.g. methyl) and the other represents: (i) C1-3 alkyl optionally substituted by one or more fluoro atoms (e.g. methyl, —CH2CF3); (ii) a 5-membered heteroaryl group containing one or two nitrogen atoms (e.g. pyrazolyl, such as 4-pyrazolyl) optionally substituted by one or two substituents selected from C1-3 alkyl, haloC1-3alkyl and —(CH2)n1-heterocyclyl (for instance where nl represents 1, and the heterocycle is a 5-membered oxygen containing ring, e.g. tetrahydrofuran, optionally containing a —CH2— bridge): (iii) aryl (e.g. phenyl) optionally substituted by one or more substituents selected from halo, C1-3 alkyl and —OC1-3 alkyl: (iv) -X1a-Y1a, in which X1a represents a direct bond or —CH2, and Y1a represents C3-6 cycloalkyl such as unsubstituted cyclopentyl and unsubstituted cyclopropyl: (v) -X1b-Y1b, in which e.g. X1b represents a direct bond, and Y1b represents 5- or 6-membered heterocyclyl containing one or two (e.g. one) nitrogen atom(s) (so forming e.g. pyrrolidine, such as 3-pyrrolidine), and which group is optionally substituted with one or two (e.g. one) substituent(s) selected from halo, C1-3 alkyl and —OC1-3 alkyl (e.g. the heterocyclyl may be substituted by methyl).
In an embodiment, R3a and R3b are linked together to form a 4-6 membered ring (optionally containing one further heteroatom, e.g. oxygen, so forming e.g. morpholinyl), optionally containing a bridge (e.g. a —CH2— bridge), optionally containing a further 3- to 6-membered spiro cycle and/or optionally being fused to a further 3- to 6-membered ring (but wherein in all cases, the maximum number of atoms of the first ring plus the bridge, spiro cycle or fused ring is 10). Such ring system is optionally substituted with one or more substituents selected from halo (e.g. fluoro), C1-3 alkyl, —OH, —OC1-3 alkyl, —N(C1-3 alkyl), and C1-3 alkyoxyC1-3 alkyl.
In an embodiment R4 represents hydrogen, halo, C1-3 alkyl or C3-6 cycloalkyl. In a particular embodiment R4 represents hydrogen, bromo or cyclopropyl. In a certain embodiment, R4 represents hydrogen.
The names of the compounds of the present invention were generated according to the nomenclature rules agreed upon by the Chemical Abstracts Service (CAS) using Advanced Chemical Development, Inc., software (ACD/Name product version 10.01; Build 15494, 1 Dec 2006) or according to the nomenclature rules agreed upon by the International Union of Pure and Applied Chemistry (IUPAC) using Advanced Chemical Development, Inc., software (ACD/Name product version 10.01.0.14105, October 2006). In case of tautomeric forms, the name of the depicted tautomeric form of the structure was generated. The other non-depicted tautomeric form is also included within the scope of the present invention.
In an aspect of the invention, there is provided a process for the preparation of compounds of the invention, where reference here is made to compounds of formula (I) as defined herein.
Compounds of formula (I) may be prepared by:
(i) reaction of a compound of formula (II),
or a derivative thereof, wherein R1 and R2 are as hereinbefore defined, with a compound of formula (III),
HNR3aR3b (III)
or a derivative thereof, wherein R3a and R3b are as hereinbefore defined, either under nucleophilic substitution reaction conditions, for example in the presence of a suitable base (e.g. diisopropylethylamine) in a suitable solvent such as acetonitrile and dimethylformamide, or by a Buchwald-type cross coupling reaction in the presence of a catalyst, e.g. tris(dibenzylideneacetone)dipalladium(0), a ligand, e.g. 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene and a base e.g. Na2CO3 in a suitable solvent such as 1,4-dioxane and dimethylformamide
In general, the compounds of the invention can therefore be made with reference to the procedures above. However, in the interests of versatility, further schemes are provided below in order to provide intermediate compounds of the invention. Further details are provided in the schemes below (as well as in the specific details of the experimental described hereinafter).
In this respect, Scheme 1 outlines a typical synthesis:
The compound of formula (II), wherein R1 and R2 are as hereinbefore defined, and R4 is hydrogen, can be prepared by a reaction sequence shown in Scheme 1 (above), whereby following flow conditions the acrylate ester (M1) is magnesiated by reaction with a strong and non-nucleophilic base, e.g. 2,2,6,6-tetramethylpiperidinylmagnesium chloride lithium chloride, that is quenched with an appropriate acyl chloride, wherein R2 is as hereinbefore defined, in the presence of a catalytic amount of copper(I) cyanide and lithium chloride, followed by reaction with hydrazine to give pyridazinone (M2), which is then alkylated with an appropriate alkyl haloacetate, wherein R is C1-4 alkyl, in the presence of a base, e.g. Cs2CO3, to provide ester (M3) which is then reacted with a chlorinating reagent, e.g. phosphoryl chloride, to yield intermediate (M4) which is treated under basic conditions, e.g. aqueous LiOH in THF to yield the acid intermediate (M5) followed by amidation with R1-NH2 using standard coupling conditions, e.g. HATU and a base, e.g. Hünig's base, to provide a compound of Formula (II).
Certain intermediate compounds may be commercially available, may be known in the literature, or may be obtained either by analogy with the processes described herein, or by conventional synthetic procedures, in accordance with standard techniques, from available starting materials using appropriate reagents and reaction conditions.
Certain substituents on/in final compounds of the invention or relevant intermediates may be modified one or more times, after or during the processes described above by way of methods that are well known to those skilled in the art. Examples of such methods include substitutions, reductions, oxidations, alkylations, acylations, hydrolyses, esterifications, etherifications, halogenations, nitrations or couplings.
Compounds of the invention may be isolated from their reaction mixtures using conventional techniques (e.g. recrystallisations, where possible under standard conditions).
It will be appreciated by those skilled in the art that, in the processes described above and hereinafter, the functional groups of intermediate compounds may need to be protected by protecting groups.
The need for such protection will vary depending on the nature of the remote functionality and the conditions of the preparation methods (and the need can be readily determined by one skilled in the art). Suitable amino-protecting groups include acetyl, trifluoroacetyl, t-butoxy carbonyl (BOC), benzyloxy carbonyl (CBz), 9-fluorenyl-methy leneoxy carbonyl (Fmoc) and 2,4,4-trimethylpentan-2-yl (which may be deprotected by reaction in the presence of an acid, e.g. HCl in water/alcohol (e.g. MeOH)) or the like. The need for such protection is readily determined by one skilled in the art. For example the a —C(O)O-tert-butyl ester moiety may serve as a protecting group for a —C(O)OH moiety, and hence the former may be converted to the latter for instance by reaction in the presence of a mild acid (e.g. TFA, or the like).
The protection and deprotection of functional groups may take place before or after a reaction in the above-mentioned schemes.
Protecting groups may be removed in accordance with techniques that are well known to those skilled in the art and as described hereinafter. For example, protected compounds/intermediates described herein may be converted chemically to unprotected compounds using standard deprotection techniques.
The type of chemistry involved will dictate the need, and type, of protecting groups as well as the sequence for accomplishing the synthesis.
The use of protecting groups is fully described in “Protective Groups in Organic Synthesis”, 3rd edition, T. W. Greene & P. G. M. Wutz, Wiley-Interscience (1999).
The compounds of the invention as prepared in the hereinabove described processes may be synthesized in the form of racemic mixtures of enantiomers which can be separated from one another following art-known resolution procedures. Those compounds of the invention that are obtained in racemic form may be converted into the corresponding diastereomeric salt forms by reaction with a suitable chiral acid. Said diastereomeric salt forms are subsequently separated, for example, by selective or fractional crystallization and the enantiomers are liberated therefrom by alkali. An alternative manner of separating the enantiomeric forms of the compounds of the invention involves liquid chromatography using a chiral stationary phase. Said pure stereochemically isomeric forms may also be derived from the corresponding pure stereochemically isomeric forms of the appropriate starting materials, provided that the reaction occurs stereospecifically. Preferably if a specific stereoisomer is desired, said compound will be synthesized by stereospecific methods of preparation. These methods will advantageously employ enantiomerically pure starting materials.
There is evidence for a role of NLRP3-induced IL-1 and IL-18 in the inflammatory responses occurring in connection with, or as a result of, a multitude of different disorders (Menu et al., Clinical and Experimental Immunology. 2011, 166, 1-15: Strowig et al., Nature. 2012, 481, 278-286). NLRP3 mutations have been found to be responsible for a set of rare autoinflammatory diseases known as CAPS (Ozaki et al., J. Inflammation Research. 2015, 8, 15-27: Schroder et al., Cell. 2010, 140: 821-832: Menu et al., Clinical and Experimental Immunology. 2011, 166, 1-15). CAPS are heritable diseases characterized by recurrent fever and inflammation and are comprised of three autoinflammatory disorders that form a clinical continuum. These diseases, in order of increasing severity, are familial cold autoinflammatory syndrome (FCAS), Muckle-Wells syndrome (MWS), and chronic infantile cutaneous neurological articular syndrome (CINCA: also called neonatal- onset multisystem inflammatory disease, NOMID), and all have been shown to result from gain-of- function mutations in the NLRP3 gene, which leads to increased secretion of IL-1 beta. NLRP3 has also been implicated in a number of autoinflammatory diseases, including pyogenic arthritis, pyoderma gangrenosum and acne (PAPA), Sweet's syndrome, chronic nonbacterial osteomyelitis (CNO), and acne vulgaris (Cook et al., Eur. J. Immunol., 2010, 40, 595-653).
A number of autoimmune diseases have been shown to involve NLRP3 including, in particular, multiple sclerosis, type-1 diabetes (TID), psoriasis, rheumatoid arthritis (RA), Behcet's disease, Schnitzler syndrome, macrophage activation syndrome (Braddock et al., Nat. Rev. Drug Disc. 2004, 3, 1-10; Inoue et a\., Immunology. 2013. 139, 11-18: Coll et a\.m Nat. Med. 2015, 21(3), 248-55: Scott et al., Clin. Exp. Rheumatol. 2016, 34(1), 88-93), systemic lupus erythematosus and its complications such as lupus nephritis (Lu et al., J. Immunol., 2017, 198(3), 1119-29), and systemic sclerosis (Artlett et al., Arthritis Rheum. 2011, 63(11), 3563-74). NLRP3 has also been shown to play a role in a number of lung diseases including chronic obstructive pulmonary disorder (COPD), asthma (including steroid-resistant asthma), asbestosis, and silicosis (De Nardo et al., Am. J. Pathol., 2014. 184: 42-54: Kim et al., Am. J. Respir. Crit. Care Med. 2017, 196(3), 283-97). NLRP3 has also been suggested to have a role in a number of central nervous system conditions, including Multiple Sclerosis (MS), Parkinson's disease (PD), Alzheimer's disease (AD), dementia, Huntington's disease, cerebral malaria, brain injury from pneumococcal meningitis (Walsh et al., Nature Reviews. 2014. 15, 84-97: and Dempsey et al., Brain. Behav. Immun. 2017, 61, 306-16), intracranial aneurysms (Zhang et al., J. Stroke and Cerebrovascular Dis., 2015, 24, 5, 972-9), and traumatic brain injury (Ismael et al., J. Neurotrauma., 2018, 35(11), 1294-1303). NLRP3 activity has also been shown to be involved in various metabolic diseases including type 2 diabetes (T2D) and its organ-specific complications, atherosclerosis, obesity, gout, pseudo-gout, metabolic syndrome (Wen et al., Nature Immunology. 2012, 13, 352-357: Duewell et al., Nature. 2010, 464, 1357-1361: Strowig et al., Nature. 2014, 481, 278- 286), and non-alcoholic steatohepatitis (Mridha et al., J. Hepatol. 2017, 66(5), 1037-46). A role for NLRP3 via IL-1 beta has also been suggested in atherosclerosis, myocardial infarction (van Hout et al., Eur. Heart J. 2017, 38(11), 828-36), heart failure (Sano et al., J. Am. Coll. Cardiol. 2018, 71(8), 875-66), aortic aneurysm and dissection (Wu et al., Arteriosc\er. Thromb. Vase. Biol., 2017,37(4), 694-706), and other cardiovascular events (Ridker et al., N. Engl. J. Med., 2017, 377(12), 1119-31).
Other diseases in which NLRP3 has been shown to be involved include: ocular diseases such as both wet and dry age-related macular degeneration (Doyle et al., Nature Medicine. 2012, 18, 791-798; Tarallo et al., Cell 2012, 149(4), 847-59), diabetic retinopathy (Loukovaara et al., Acta Ophthalmol., 2017, 95(8), 803-8), non-infectious uveitis and optic nerve damage (Puyang et al., Sci. Rep. 2016, 6, 20998); liver diseases including non-alcoholic steatohepatitis (NASH) and acute alcoholic hepatitis (Henao-Meija et al., Nature. 2012, 482, 179-185); inflammatory reactions in the lung and skin (Primiano et al., J. Immunol. 2016, 197(6), 2421-33) including contact hypersensitivity (such as bullous pemphigoid (Fang et al., J Dermatol Sci.
2016, 83(2), 116-23)), atopic dermatitis (Niebuhr et al., Allergy. 2014, 69(8), 1058-67), Hidradenitis suppurativa (Alikhan et al., J. Am. Acad. Dermatol., 2009 ,60(4), 539-61), and sarcoidosis (Jager et al., Am. J. Respir. Crit. Care Med., 2015, 191, A5816): inflammatory reactions in the joints (Braddock et al., Nat. Rev. Drug Disc. 2004, 3, 1-10): amyotrophic lateral sclerosis (Gugliandolo et al., Int. J. Mo\. Sci.,
2018, 19(7), E1992); cystic fibrosis (lannitti et al., Nat. Commun., 2016, 7, 10791); stroke (Walsh et al., Nature Reviews. 2014, 15, 84-97); chronic kidney disease (Granata et al., PLoS One 2015, 10(3), eoi22272): and inflammatory bowel diseases including ulcerative colitis and Crohn's disease (Braddock et al., Nat. Rev. Drug Disc. 2004, 3, 1-10; Neudecker et al., J. Exp. Med. 2017, 214(6), 1737-52: Lazaridis et al.,
Dig. Dis. Sci. 2017, 62(9), 2348-56). The NLRP3 inflammasome has been found to be activated in response to oxidative stress. NLRP3 has also been shown to be involved in inflammatory hyperalgesia (Dolunay et al., Inflammation. 2017, 40, 366-86).
Activation of the NLRP3 inflammasome has been shown to potentiate some pathogenic infections such as influenza and Leishmaniasis (Tate et al., Sci Rep., 2016, 10(6), 27912-20; Novias et al., PLOS Pathogens 2017, 13(2), e1006196).
NLRP3 has also been implicated in the pathogenesis of many cancers (Menu et al., Clinical and Experimental Immunology. 2011. 166, 1-15). For example, several previous studies have suggested a role for IL-1 beta in cancer invasiveness, growth and metastasis, and inhibition of IL-1 beta with canakinumab has been shown to reduce the incidence of lung cancer and total cancer mortality in a randomised, double-blind, placebo-controlled trial (Ridker et al., Lancet., 2017, 390(10105), 1833-42). Inhibition of the NLRP3 inflammasome or IL-1 beta has also been shown to inhibit the proliferation and migration of lung cancer cells in vitro (Wang et al., Onco Rep., 2016, 35(4), 2053-64). A role for the NLRP3 inflammasome has been suggested in myelodysplastic syndromes, myelofibrosis and other myeloproliferative neoplasms, and acute myeloid leukemia (AML) (Basiorka et al., Blood. 2016, 128(25), 2960-75.) and also in the carcinogenesis of various other cancers including glioma (Li et al., Am. J. Cancer Res. 2015, 5(1), 442-9), inflammation- induced tumors (Allen et al., J. Exp. Med. 2010, 207(5), 1045-56; Hu et al., PNAS., 2010, 107(50), 21635-40), multiple myeloma (Li et al., Hematology. 2016 21(3), 144-51), and squamous cell carcinoma of the head and neck (Huang et al., J. Exp. Clin. Cancer Res., 2017, 36(1), 116). Activation of the NLRP3 inflammasome has also been shown to mediate chemoresistance of tumor cells to 5-Fluorouracil (Feng et al., J. Exp. Clin. Cancer Res., 2017, 36(1), 81), and activation of NLRP3 inflammasome in peripheral nerve contributes to chemotherapy-induced neuropathic pain (Jia et al., Mol. Pain., 2017, 13, 1-11). NLRP3 has also been shown to be required for the efficient control of viruses, bacteria, and fungi.
The activation of NLRP3 leads to cell pyroptosis and this feature plays an important part in the manifestation of clinical disease (Yan-gang et al., Cell Death and Disease. 2017, 8(2), 2579; Alexander et al., Hepatology. 2014, 59(3), 898-910;
Baldwin et al., J. Med. Chem., 2016, 59(5), 1691- 1710: Ozaki et a., J. Inflammation Research. 2015, 8, 15-27: Zhen et a., Neuroimmunology Neuroinflammation. 2014, 1(2), 60-65: Mattia et a., J. Med. Chem., 2014, 57(24), 10366-82: Satoh et al., Cell Death and Disease. 2013, 4, 644). Therefore, it is anticipated that inhibitors of NLRP3 will block pyroptosis, as well as the release of pro-inflammatory cytokines (e.g. IL-1 beta) from the cell.
Hence, the compounds of the invention, as described herein (e.g. in any of the embodiments described herein, including by the examples, and/or in any of the forms described herein, e.g. in a salt form or free form, etc) exhibit valuable pharmacological properties, e.g. NLRP3 inhibiting properties on the NLRP3 inflammasome pathway e.g. as indicated in vitro tests as provided herein, and are therefore indicated for therapy or for use as research chemicals, e.g. as tool compounds. Compounds of the invention may be useful in the treatment of an indication selected from: inflammasome-related diseases/disorders, immune diseases, inflammatory diseases, auto-immune diseases, or auto-inflammatory diseases, for example, of diseases, disorders or conditions in which NLRP3 signaling contributes to the pathology, and/or symptoms, and/or progression, and which may be responsive to NLRP3 inhibition and which may be treated or prevented, according to any of the methods/uses described herein, e.g. by use or administration of a compound of the invention, and, hence, in an embodiment, such indications may include:
More specifically the compounds of the invention may be useful in the treatment of an indication selected from: inflammasome-related diseases/disorders, immune diseases, inflammatory diseases, auto-immune diseases, or auto-inflammatory diseases, for example, autoinflammatory fever syndromes (e.g., cryopyrin-associated periodic syndrome), sickle cell disease, systemic lupus erythematosus (SLE), liver related diseases/disorders (e.g. chronic liver disease, viral hepatitis, non-alcoholic steatohepatitis (NASH), alcoholic steatohepatitis, and alcoholic liver disease), inflammatory arthritis related disorders (e.g. gout, pseudogout (chondrocalcinosis), osteoarthritis, rheumatoid arthritis, arthropathy e.g. acute, chronic), kidney related diseases (e.g. hyperoxaluria, lupus nephritis. Type I/Type II diabetes and related complications (e.g. nephropathy, retinopathy), hypertensive nephropathy. hemodialysis related inflammation), neuroinflammation-related diseases (e.g. multiple sclerosis, brain infection, acute injury, neurodegenerative diseases, Alzheimer's disease), cardiovascular/metabolic diseases/disorders (e.g. cardiovascular risk reduction (CvRR), hypertension, atherosclerosis. Type I and Type II diabetes and related complications, peripheral artery disease (PAD), acute heart failure), inflammatory skin diseases (e.g. hidradenitis suppurativa, acne), wound healing and scar formation, asthma, sarcoidosis, age-related macular degeneration, and cancer related diseases/disorders (e.g. colon cancer, lung cancer, myeloproliferative neoplasms, leukemias, myelodysplastic syndromes (MOS), myelofibrosis). In particular, autoinflammatory fever syndromes (e.g. CAPS), sickle cell disease. Type I/Type II diabetes and related complications (e.g. nephropathy, retinopathy), hyperoxaluria, gout, pseudogout (chondrocalcinosis), chronic liver disease. NASH, neuroinflammation-related disorders (e.g. multiple sclerosis, brain infection, acute injury, neurodegenerative diseases, Alzheimer's disease), atherosclerosis and cardiovascular risk (e.g. cardiovascular risk reduction (CvRR), hypertension), hidradenitis suppurativa, wound healing and scar formation, and cancer (e.g. colon cancer, lung cancer, myeloproliferative neoplasms, leukemias, myelodysplastic syndromes (MOS), myelofibrosis).
In particular, compounds of the invention, may be useful in the treatment of a disease or disorder selected from autoinflammatory fever syndromes (e.g. CAPS), sickle cell disease, Type I/Type II diabetes and related complications (e.g. nephropathy, retinopathy), hyperoxaluria, gout, pseudogout (chondrocalcinosis), chronic liver disease, NASH, neuroinflammation-related disorders (e.g. multiple sclerosis, brain infection, acute injury, neurodegenerative diseases, Alzheimer's disease), atherosclerosis and cardiovascular risk (e.g. cardiovascular risk reduction (CvRR), hypertension), hidradenitis suppurativa, wound healing and scar formation, and cancer (e.g. colon cancer, lung cancer, myeloproliferative neoplasms, leukemias, myelodysplastic syndromes (MOS), myelofibrosis). Thus, as a further aspect, the present invention provides the use of a compound of the invention (hence, including a compound as defined by any of the embodiments/forms/examples herein) in therapy. In a further embodiment, the therapy is selected from a disease, which may be treated by inhibition of NLRP3 inflammasome. In another embodiment, the disease is as defined in any of the lists herein. Hence, there is provided any one of the compounds of the invention described herein (including any of the embodiments/forms/examples) for use in the treatment of any of the diseases or disorders described herein (e.g. as described in the aforementioned lists).
In an embodiment, the invention also relates to a composition comprising a pharmaceutically acceptable carrier and, as active ingredient, a therapeutically effective amount of a compound of the invention. The compounds of the invention may be formulated into various pharmaceutical forms for administration purposes. As appropriate compositions there may be cited all compositions usually employed for systemically administering drugs. To prepare the pharmaceutical compositions of this invention, an effective amount of the particular compound, optionally in salt form, as the active ingredient is combined in intimate admixture with a pharmaceutically acceptable carrier, which carrier may take a wide variety of forms depending on the form of preparation desired for administration. These pharmaceutical compositions are desirable in unitary dosage form suitable, in particular, for administration orally or by parenteral injection. For example, in preparing the compositions in oral dosage form, any of the usual pharmaceutical media may be employed such as, for example, water, glycols, oils, alcohols and the like in the case of oral liquid preparations such as suspensions, syrups, elixirs, emulsions and solutions: or solid carriers such as starches, sugars, kaolin, diluents, lubricants, binders, disintegrating agents and the like in the case of powders, pills, capsules and tablets. Because of their ease in administration, tablets and capsules represent the most advantageous oral dosage unit forms in which case solid pharmaceutical carriers are obviously employed. For parenteral compositions, the carrier will usually comprise sterile water, at least in large part, though other ingredients, for example, to aid solubility, may be included. Injectable solutions, for example, may be prepared in which the carrier comprises saline solution, glucose solution or a mixture of saline and glucose solution. Injectable suspensions may also be prepared in which case appropriate liquid carriers, suspending agents and the like may be employed. Also included are solid form preparations which are intended to be converted, shortly before use, to liquid form preparations.
In an embodiment, and depending on the mode of administration, the pharmaceutical composition will preferably comprise from 0.05 to 99% by weight, more preferably from 0.1 to 70% by weight, even more preferably from 0.1 to 50% by weight of the active ingredient(s), and, from 1 to 99.95% by weight, more preferably from 30 to 99.9% by weight, even more preferably from 50 to 99.9% by weight of a pharmaceutically acceptable carrier, all percentages being based on the total weight of the composition.
The pharmaceutical composition may additionally contain various other ingredients known in the art, for example, a lubricant, stabilising agent, buffering agent, emulsifying agent, viscosity-regulating agent, surfactant, preservative, flavouring or colorant.
It is especially advantageous to formulate the aforementioned pharmaceutical compositions in unit dosage form for ease of administration and uniformity of dosage. Unit dosage form as used herein refers to physically discrete units suitable as unitary dosages, each unit containing a predetermined quantity of active ingredient calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. Examples of such unit dosage forms are tablets (including scored or coated tablets), capsules, pills, powder packets, wafers, suppositories, injectable solutions or suspensions and the like, and segregated multiples thereof.
The daily dosage of the compound according to the invention will, of course, vary with the compound employed, the mode of administration, the treatment desired and the mycobacterial disease indicated. However, in general, satisfactory results will be obtained when the compound according to the invention is administered at a daily dosage not exceeding 1 gram. e.g. in the range from 10 to 50 mg/kg body weight.
In an embodiment, there is provided a combination comprising a therapeutically effective amount of a compound of the invention, according to any one of the embodiments described herein, and another therapeutic agent (including one or more therapeutic agents). In a further embodiment, there is provided such a combination wherein the other therapeutic agent is selected from (and where there is more than one therapeutic agent, each is independently selected from): farnesoid X receptor (FXR) agonists: anti-steatotics: anti-fibrotics: JAK inhibitors: checkpoint inhibitors including anti-PDI inhibitors, anti-LAG-3 inhibitors, anti-TIM-3 inhibitors, or anti-POL 1 inhibitors: chemotherapy, radiation therapy and surgical procedures: urate-lowering therapies: anabolics and cartilage regenerative therapy: blockade of IL-17: complement inhibitors: Bruton's tyrosine Kinase inhibitors (BTK inhibitors): Toll Like receptor inhibitors (TLR7/8 inhibitors): CAR-T therapy: anti-hypertensive agents: cholesterol lowering agents: leukotriene A4 hydrolase (LTAH4) inhibitors: SGLT2 inhibitors: 132-agonists: anti-inflammatory agents: nonsteroidal anti-inflammatory drugs (“NSAIDs”): acetylsalicylic acid drugs (ASA) including aspirin: paracetamol: regenerative therapy treatments: cystic fibrosis treatments: or atherosclerotic treatment. In a further embodiment, there is also provided such (a) combination(s) for use as described herein in respect of compounds of the invention, e.g. for use in the treatment of a disease or disorder in which the NLRP3 signaling contributes to the pathology. and/or symptoms, and/or progression, of said disease/disorder, or, a disease or disorder associated with NLRP3 activity (including NLRP3 inflammasome activity), including inhibiting NLRP3 inflammasome activity, and in this respect the specific disease/disorder mentioned herein apply equally here. There may also be provided methods as described herein in respect of compounds of the invention, but wherein the method comprises administering a therapeutically effective amount of such combination (and, in an embodiment, such method may be to treat a disease or disorder mentioned herein in the context of inhibiting NLRP3 inflammasome activity). The combinations mentioned herein may be in a single preparation or they may be formulated in separate preparations so that they can be administered simultaneously, separately or sequentially. Thus, in an embodiment, the present invention also relates to a combination product containing (a) a compound according to the invention, according to any one of the embodiments described herein, and (b) one or more other therapeutic agents (where such therapeutic agents are as described herein), as a combined preparation for simultaneous, separate or sequential use in the treatment of a disease or disorder associated with inhibiting NLRP3 inflammasome activity (and where the disease or disorder may be any one of those described herein), for instance, in an embodiment, the combination may be a kit of parts. Such combinations may be referred to as “pharmaceutical combinations”. The route of administration for a compound of the invention as a component of a combination may be the same or different to the one or more other therapeutic agent(s) with which it is combined. The other therapeutic agent is, for example, a chemical compound, peptide, antibody, antibody fragment or nucleic acid, which is therapeutically active or enhances the therapeutic activity when administered to a patient in combination with a compound of the invention.
The weight ratio of (a) the compound according to the invention and (b) the other therapeutic agent(s) when given as a combination may be determined by the person skilled in the art. Said ratio and the exact dosage and frequency of administration depends on the particular compound according to the invention and the other antibacterial agent(s) used, the particular condition being treated, the severity of the condition being treated, the age, weight, gender, diet, time of administration and general physical condition of the particular patient, the mode of administration as well as other medication the individual may be taking, as is well known to those skilled in the art. Furthermore, it is evident that the effective daily amount may be lowered or increased depending on the response of the treated subject and/or depending on the evaluation of the physician prescribing the compounds of the instant invention. A particular weight ratio for the present compound of the invention and another antibacterial agent may range from 1/10 to 10/1, more in particular from 1/5 to 5/1, even more in particular from 1/3 to 3/1.
The pharmaceutical composition or combination of the present invention can be in unit dosage of about 1-1000 mg of active ingredient(s) for a subject of about 50-70 kg, or about 1-500 mg, or about 1-250 mg, or about 1-150 mg. or about 1-100 mg, or about 1-50 mg of active ingredients. The therapeutically effective dosage of a compound, the pharmaceutical composition, or the combinations thereof, is dependent on the species of the subject, the body weight, age and individual condition, the disorder or disease or the severity thereof being treated. A physician, clinician or veterinarian of ordinary skill can readily determine the effective amount of each of the active ingredients necessary to prevent, treat or inhibit the progress of the disorder or disease.
The above-cited dosage properties are demonstrable in vitro and in vivo tests using advantageously mammals, e.g., mice, rats, dogs, monkeys or isolated organs, tissues and preparations thereof. The compounds of the present invention can be applied in vitro in the form of solutions, e.g., aqueous solutions, and in vivo either enterally, parenterally, advantageously intravenously, e.g., as a suspension or in aqueous solution. The dosage in vitro may range between about 10-3 molar and 10-9 molar concentrations. A therapeutically effective amount in vivo may range depending on the route of administration, between about 0.1-500 mg/kg, or between about 1-100 mg/kg.
As used herein, term “pharmaceutical composition” refers to a compound of the invention, or a pharmaceutically acceptable salt thereof, together with at least one pharmaceutically acceptable carrier, in a form suitable for oral or parenteral administration.
As used herein, the term “pharmaceutically acceptable carrier” refers to a substance useful in the preparation or use of a pharmaceutical composition and includes, for example, suitable diluents, solvents, dispersion media, surfactants, antioxidants, preservatives, isotonic agents, buffering agents, emulsifiers, absorption delaying agents, salts, drug stabilizers, binders, excipients, disintegration agents, lubricants, wetting agents, sweetening agents, flavoring agents, dyes, and combinations thereof, as would be known to those skilled in the art (see, for example, Remington The Science and Practice of Pharmacy, 22nd Ed. Pharmaceutical Press, 2013, pp. 1049-1070).
The term “subject” as used herein, refers to an animal, preferably a mammal, most preferably a human, for example who is or has been the object of treatment, observation or experiment.
The term “therapeutically effective amount” as used herein, means that amount of compound of the invention (including, where applicable, form, composition, combination comprising such compound of the invention) elicits the biological or medicinal response of a subject, for example, reduction or inhibition of an enzyme or a protein activity, or ameliorate symptoms, alleviate conditions, slow or delay disease progression, or prevent a disease, etc. In one non-limiting embodiment, the term “a therapeutically effective amount” refers to the amount of the compound of the present invention that, when administered to a subject, is effective to (1) at least partially alleviate, inhibit, prevent and/or ameliorate a condition, or a disorder or a disease (i) mediated by NLRP3, or (ii) associated with NLRP3 activity, or (iii) characterised by activity (normal or abnormal) of NLRP3: or (2) reduce or inhibit the activity of NLRP3: or (3) reduce or inhibit the expression of NLRP3. In another non-limiting embodiment, the term “a therapeutically effective amount” refers to the amount of the compound of the present invention that, when administered to a cell, or a tissue, or a non-cellular biological material, or a medium, is effective to at least partially reduce or inhibit the activity of NLRP3: or at least partially reduce or inhibit the expression of NLRP3.
As used herein, the term “inhibit”, “inhibition” or “inhibiting” refers to the reduction or suppression of a given condition, symptom, or disorder, or disease, or a significant decrease in the baseline activity of a biological activity or process. Specifically, inhibiting NLRP3 or inhibiting NLRP3 inflammasome pathway comprises reducing the ability of NLRP3 or NLRP3 inflammasome pathway to induce the production of IL-1 and/or IL-18. This can be achieved by mechanisms including, but not limited to, inactivating, destabilizing, and/or altering distribution of NLRP3.
As used herein, the term “NLRP3” is meant to include, without limitation, nucleic acids, polynucleotides, oligonucleotides, sense and anti-sense polynucleotide strands, complementary sequences, peptides, polypeptides, proteins, homologous and/or orthologous NLRP molecules, isoforms, precursors, mutants, variants, derivatives, splice variants, alleles, different species, and active fragments thereof.
As used herein, the term “treat”, “treating” or “treatment” of any disease or disorder refers to alleviating or ameliorating the disease or disorder (i.e., slowing or arresting the development of the disease or at least one of the clinical symptoms thereof): or alleviating or ameliorating at least one physical parameter or biomarker associated with the disease or disorder, including those which may not be discernible to the patient.
As used herein, the term “prevent”. “preventing” or “prevention” of any disease or disorder refers to the prophylactic treatment of the disease or disorder; or delaying the onset or progression of the disease or disorder.
As used herein, a subject is “in need of” a treatment if such subject would benefit biologically, medically or in quality of life from such treatment.
“Combination” refers to either a fixed combination in one dosage unit form, or a combined administration where a compound of the present invention and a combination partner (e.g. another drug as explained below, also referred to as “therapeutic agent” or “co-agent”) may be administered independently at the same time or separately within time intervals. The single components may be packaged in a kit or separately. One or both of the components (e.g. powders or liquids) may be reconstituted or diluted to a desired dose prior to administration. The terms “co-administration” or “combined administration” or the like as utilized herein are meant to encompass administration of the selected combination partner to a single subject in need thereof (e.g. a patient), and are intended to include treatment regimens in which the agents are not necessarily administered by the same route of administration or at the same time.
The term “pharmaceutical combination” as used herein means a product that results from the mixing or combining of more than one therapeutic agent and includes both fixed and non-fixed combinations of the therapeutic agents. The term “pharmaceutical combination” as used herein refers to either a fixed combination in one dosage unit form, or non-fixed combination or a kit of parts for the combined administration where two or more therapeutic agents may be administered independently at the same time or separately within time intervals. The term “fixed combination” means that the therapeutic agents, e.g. a compound of the present invention and a combination partner, are both administered to a patient simultaneously in the form of a single entity or dosage. The term “non-fixed combination” means that the therapeutic agents, e.g. a compound of the present invention and a combination partner, are both administered to a patient as separate entities either simultaneously, concurrently or sequentially with no specific time limits, wherein such administration provides therapeutically effective levels of the two compounds in the body of the patient. The latter also applies to cocktail therapy, e.g. the administration of three or more therapeutic agents.
The term “combination therapy” refers to the administration of two or more therapeutic agents to treat a therapeutic condition or disorder described in the present disclosure. Such administration encompasses co-administration of these therapeutic agents in a substantially simultaneous manner, such as in a single capsule having a fixed ratio of active ingredients. Alternatively, such administration encompasses co-administration in multiple, or in separate containers (e.g. tablets, capsules, powders, and liquids) for each active ingredient. Powders and/or liquids may be reconstituted or diluted to a desired dose prior to administration. In addition, such administration also encompasses use of each type of therapeutic agent in a sequential manner, either at approximately the same time or at different times. In either case, the treatment regimen will provide beneficial effects of the drug combination in treating the conditions or disorders described herein.
In an embodiment, there is provided a pharmaceutical composition comprising a therapeutically effective amount of a compound of the invention, according to any one of the embodiments described herein, and a pharmaceutically acceptable carrier (including one or more pharmaceutically acceptable carriers).
In an embodiment, there is provided a compound of the invention, according to any one of the embodiments described herein, for use as a medicament.
In an embodiment, there is provided a compound of the invention, according to any one of the embodiments described herein (and/or pharmaceutical compositions comprising such compound of the invention, according to any one of the embodiment described herein) for use: in the treatment of a disease or disorder associated with NLRP3 activity (including inflammasome activity); in the treatment of a disease or disorder in which the NLRP3 signalling contributes to the pathology, and/or symptoms, and/or progression, of said disease/disorder: in inhibiting NLRP3 inflammasome activity (including in a subject in need thereof): and/or as an NLRP3 inhibitor.
In an embodiment, there is provided a use of compounds of the invention. according to any one of the embodiments described herein (and/or pharmaceutical compositions comprising such compound of the invention, according to any one of the embodiment described herein): in the treatment of a disease or disorder associated with NLRP3 activity (including inflammasome activity): in the treatment of a disease or disorder in which the NLRP3 signalling contributes to the pathology. and/or symptoms. and/or progression, of said disease/disorder: in inhibiting NLRP3 inflammasome activity (including in a subject in need thereof); and/or as an NLRP3 inhibitor.
In an embodiment, there is provided use of compounds of the invention. according to any one of the embodiments described herein (and/or pharmaceutical compositions comprising such compound of the invention, according to any one of the embodiment described herein), in the manufacture of a medicament for: the treatment of a disease or disorder associated with NLRP3 activity (including inflammasome activity): the treatment of a disease or disorder in which the NLRP3 signalling contributes to the pathology, and/or symptoms, and/or progression, of said disease/disorder: and/or inhibiting NLRP3 inflammasome activity (including in a subject in need thereof).
In an embodiment, there is provided a method of treating a disease or disorder in which the NLRP3 signalling contributes to the pathology, and/or symptoms, and/or progression, of said disease/disorder, comprising administering a therapeutically effective amount of a compound of the invention, according to any one of the embodiments described herein (and/or pharmaceutical compositions comprising such compound of the invention, according to any one of the embodiment described herein). for instance to a subject (in need thereof). In a further embodiment, there is provided a method of inhibiting the NLRP3 inflammasome activity in a subject (in need thereof), the method comprising administering to the subject in need thereof a therapeutically effective amount of a compound of the invention, according to any one of the embodiments described herein (and/or pharmaceutical compositions comprising such compound of the invention, according to any one of the embodiment described herein).
In all relevant embodiment of the invention, where a disease or disorder is mentioned (e.g. hereinabove), for instance a disease or disorder in which the NLRP3 signalling contributes to the pathology, and/or symptoms, and/or progression, of said disease/disorder, or, a disease or disorder associated with NLRP3 activity (including NLRP3 inflammasome activity), including inhibiting NLRP3 inflammasome activity. then such disease may include inflammasome-related diseases or disorders, immune diseases, inflammatory diseases, auto-immune diseases, or auto-inflammatory diseases. In a further embodiment, such disease or disorder may include autoinflammatory fever syndromes (e.g. cryopyrin-associated periodic syndrome), liver related diseases/disorders (e.g. chronic liver disease, viral hepatitis, non-alcoholic steatohepatitis (NASH), alcoholic steatohepatitis, and alcoholic liver disease). inflammatory arthritis related disorders (e.g. gout, pseudogout (chondrocalcinosis), osteoarthritis, rheumatoid arthritis, arthropathy e.g. acute, chronic), kidney related diseases (e.g. hyperoxaluria, lupus nephritis. Type I/Type II diabetes and related complications (e.g. nephropathy, retinopathy), hypertensive nephropathy. hemodialysis related inflammation), neuroinflammation-related diseases (e.g. multiple sclerosis, brain infection, acute injury, neurodegenerative diseases. Alzheimer's disease), cardiovascular/metabolic diseases/disorders (e.g. cardiovascular risk reduction (CvRR), hypertension, atherosclerosis. Type I and Type II diabetes and related complications, peripheral artery disease (PAD), acute heart failure). inflammatory skin diseases (e.g. hidradenitis suppurativa, acne), wound healing and scar formation, asthma, sarcoidosis, age-related macular degeneration, and cancer related diseases/disorders (e.g. colon cancer, lung cancer, myeloproliferative neoplasms, leukaemia, myelodysplastic syndromes (MOS), myelofibrosis). In a particular aspect, such disease or disorder is selected from autoinflammatory fever syndromes (e.g. CAPS), sickle cell disease. Type I/Type II diabetes and related complications (e.g. nephropathy, retinopathy), hyperoxaluria, gout, pseudogout (chondrocalcinosis), chronic liver disease. NASH, neuroinflammation-related disorders (e.g. multiple sclerosis, brain infection, acute injury, neurodegenerative diseases. Alzheimer's disease), atherosclerosis and cardiovascular risk (e.g. cardiovascular risk reduction (CvRR), hypertension), hidradenitis suppurativa, wound healing and scar formation, and cancer (e.g. colon cancer, lung cancer. myeloproliferative neoplasms, leukemias, myelodysplastic syndromes (MOS). myelofibrosis). In a particular embodiment, the disease or disorder associated with inhibition of NLRP3 inflammasome activity is selected from inflammasome related diseases and disorders, immune diseases, inflammatory diseases, auto-immune diseases, auto-inflammatory fever syndromes, cryopyrin-associated periodic syndrome, chronic liver disease, viral hepatitis, non-alcoholic steatohepatitis, alcoholic steatohepatitis, alcoholic liver disease, inflammatory arthritis related disorders, gout, chondrocalcinosis, osteoarthritis, rheumatoid arthritis, chronic arthropathy, acute arthropathy, kidney related disease, hyperoxaluria, lupus nephritis. Type I and Type II diabetes, nephropathy, retinopathy, hypertensive nephropathy, hemodialysis related inflammation, neuroinflammation-related diseases, multiple sclerosis, brain infection, acute injury, neurodegenerative diseases. Alzheimer's disease, cardiovascular diseases, metabolic diseases, cardiovascular risk reduction, hypertension, atherosclerosis. peripheral artery disease, acute heart failure, inflammatory skin diseases, acne, wound healing and scar formation, asthma, sarcoidosis, age-related macular degeneration. colon cancer, lung cancer, myeloproliferative neoplasms, leukemias, myelodysplastic syndromes and myelofibrosis.
In an embodiment, there is provided a combination comprising a therapeutically effective amount of a compound of the invention, according to any one of the embodiments described herein, and another therapeutic agent (including one or more therapeutic agents). In a further embodiment, there is provided such a combination wherein the other therapeutic agent is selected from (and where there is more than one therapeutic agent, each is independently selected from): farnesoid X receptor (FXR) 20) agonists: anti-steatotics: anti-fibrotics: JAK inhibitors: checkpoint inhibitors including anti-PDI inhibitors, anti-LAG-3 inhibitors, anti-TIM-3 inhibitors, or anti-POL 1 inhibitors: chemotherapy,radiation therapy and surgical procedures: urate-lowering therapies: anabolics and cartilage regenerative therapy: blockade of IL-17: complement inhibitors: Bruton's tyrosine Kinase inhibitors (BTK inhibitors): Toll Like receptor inhibitors (TLR7/8 inhibitors): CAR-T therapy: anti-hypertensive agents: cholesterol lowering agents: leukotriene A4 hydrolase (LTAH4) inhibitors: SGLT2 inhibitors: 132-agonists: anti-inflammatory agents: nonsteroidal anti-inflammatory drugs (“NSAIDs”): acetylsalicylic acid drugs (ASA) including aspirin: paracetamol: regenerative therapy treatments: cystic fibrosis treatments: or atherosclerotic treatment. In a further embodiment, there is also provided such (a) combination(s) for use as described herein in respect of compounds of the invention, e.g. for use in the treatment of a disease or disorder in which the NLRP3 signaling contributes to the pathology. and/or symptoms, and/or progression, of said disease/disorder, or, a disease or disorder associated with NLRP3 activity (including NLRP3 inflammasome activity), including inhibiting NLRP3 inflammasome activity. and in this respect the specific disease/disorder mentioned herein apply equally here. There may also be provided methods as described herein in respect of compounds of the invention, but wherein the method comprises administering a therapeutically effective amount of such combination (and, in an embodiment, such method may be to treat a disease or disorder mentioned herein in the context of inhibiting NLRP3 inflammasome activity). The combinations mentioned herein may be in a single preparation or they may be formulated in separate preparations so that they can be administered simultaneously, separately or sequentially. Thus, in an embodiment, the present invention also relates to a combination product containing (a) a compound according to the invention, according to any one of the embodiments described herein, and (b) one or more other therapeutic agents (where such therapeutic agents are as described herein), as a combined preparation for simultaneous, separate or sequential use in the treatment of a disease or disorder associated with inhibiting NLRP3 inflammasome activity (and where the disease or disorder may be any one of those described herein).
Compounds of the invention (including forms and compositions/combinations comprising compounds of the invention) may have the advantage that they may be more efficacious than, be less toxic than, be longer acting than, be more potent than, produce fewer side effects than, be more easily absorbed than, and/or have a better pharmacokinetic profile (e.g. higher oral bioavailability and/or lower clearance) than, and/or have other useful pharmacological, physical, or chemical properties over, compounds known in the prior art, whether for use in the above-stated indications or otherwise.
For instance, compounds of the invention may have the advantage that they have a good or an improved thermodynamic solubility (e.g. compared to compounds known in the prior art: and for instance as determined by a known method and/or a method described herein). Compounds of the invention may have the advantage that they will block pyroptosis, as well as the release of pro-inflammatory cytokines (e.g. IL-1B) from the cell. Compounds of the invention may also have the advantage that they avoid side-effects, for instance as compared to compounds of the prior art, which may be due to selectivity of NLRP3 inhibition. Compounds of the invention may also have the advantage that they have good or improved in vivo pharmacokinetics and oral bioavailability. They may also have the advantage that they have good or improved in vivo efficacy. Specifically, compounds of the invention may also have advantages over prior art compounds when compared in the tests outlined hereinafter (e.g. in Examples C and D).
The compounds according to the invention can generally be prepared by a succession of steps, each of which may be known to the skilled person or described herein,
It is evident that in the foregoing and in the following reactions, the reaction products may be isolated from the reaction medium and, if necessary, further purified according to methodologies generally known in the art, such as extraction, crystallization and chromatography. It is further evident that reaction products that exist in more than one enantiomeric form, may be isolated from their mixture by known techniques, in particular preparative chromatography, such as preparative HPLC, chiral chromatography. Individual diastereoisomers or individual enantiomers can also be obtained by Supercritical Fluid Chromatography (SFC).
The starting materials and the intermediates are compounds that are either commercially available or may be prepared according to conventional reaction procedures generally known in the art.
LC-MS (LIQUID CHROMATOGRAPHY/MASS SPECTROMETRY)
The High Performance Liquid Chromatography (HPLC) measurement was performed using a LC pump, a diode-array (DAD) or a UV detector and a column as specified in the respective methods. If necessary, additional detectors were included (see table of methods below).
Flow from the column was brought to the Mass Spectrometer (MS) which was configured with an atmospheric pressure ion source. It is within the knowledge of the skilled person to set the tune parameters (e.g. scanning range, dwell time . . . ) in order to obtain ions allowing the identification of the compound's nominal monoisotopic molecular weight (MW). Data acquisition was performed with appropriate software. Compounds are described by their experimental retention times (Rt) and ions. If not specified differently in the table of data, the reported molecular ion corresponds to the [M+H]+ (protonated molecule) and/or [M-H]−(deprotonated molecule). In case the compound was not directly ionizable the type of adduct is specified (i.e. [M+NH4]+, [M+HCOO]−, etc . . . ). For molecules with multiple isotopic patterns (Br, Cl..), the reported value is the one obtained for the lowest isotope mass. All results were obtained with experimental uncertainties that are commonly associated with the method used. Hereinafter. “SQD” means Single Quadrupole Detector, “MSD” Mass Selective Detector, “RT” room temperature. “BEH” bridged ethylsiloxane/silica hybrid. “DAD” Diode Array Detector, “HSS” High Strength silica.
For a number of compounds. 1H NMR spectra were recorded on a Bruker Avance III spectrometer operating at 300 or 400 MHZ, on a Bruker Avance III-HD operating at 400 MHZ, on a Bruker Avance NEO spectrometer operating at 400 MHZ, on a Bruker Avance Neo spectrometer operating at 500 MHZ, or on a Bruker Avance 600 spectrometer operating at 600 MHZ, using CHLOROFORM-d (deuterated chloroform, CDCl3), DMSO-d6 (deuterated DMSO, dimethyl-d6 sulfoxide), METHANOL-d4 (deuterated methanol), BENZENE-d6 (deuterated benzene, C6D6) or ACETONE-d6 (deuterated acetone, (CD3)2CO) as solvents. Chemical shifts (δ) are reported in parts per million (ppm) relative to tetramethylsilane (TMS), which was used as internal standard.
Values are either peak values or melt ranges, and are obtained with experimental uncertainties that are commonly associated with this analytical method.
Method A: For a number of compounds, melting points were determined in open capillary tubes on a Mettler Toledo MP50. Melting points were measured with a temperature gradient of 10 ° C/minute. Maximum temperature was 300 ° C. The melting point data was read from a digital display and checked from a video recording system Method B: For a number of compounds, melting points were determined with a DSC823e (Mettler Toledo) apparatus. Melting points were measured with a temperature gradient of 10 ° C/minute. Standard maximum temperature was 300 ° C.
Hereinafter, the term “m.p.” means melting point, “aq.” means aqueous, “r.m.” means reaction mixture, “rt” means room temperature, ‘DIPEA’ means N,N-diiso-propy lethy lamine, “DIPE” means diisopropylether, ‘THF’ means tetrahydrofuran, ‘DMF’ means dimethylformamide, ‘DCM’ means dichloromethane, “EtOH” means ethanol ‘EtOAc’ means ethyl acetate, “AcOH” means acetic acid, “iPrOH” means isopropanol, “iPrNH2” means isopropylamine, “MeCN” or “ACN” means acetonitrile, “MeOH” means methanol, “Pd(OAc)2” means palladium(II)diacetate, “rac” means racemic, ‘sat.’ means saturated, ‘SFC’ means supercritical fluid chromatography, ‘SFC-MS’ means supercritical fluid chromatography/mass spectrometry, “LC-MS” means liquid chromatography/mass spectrometry, “GCMS” means gas chromatography/mass spectrometry, “HPLC” means high-performance liquid chromatography, “RP” means reversed phase, “UPLC” means ultra-performance liquid chromatography, “Rt” (or “RT”) means retention time (in minutes), “[M+H]+” means the protonated mass of the free base of the compound, “DAST” means diethylaminosulfur trifluoride, “DMTMM” means 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride, “HATU” means O)-(7-azabenzotriazol-1-yl)-N,N,N′, N′-tetramethyluronium hexafluorophosphate (1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate), “Xantphos” means (9,9-dimethyl-9H-xanthene-4,5-diyl)bis[diphenylphosphine], “TBAT” means tetrabutyl ammonium triphenyldifluorosilicate, “TFA” means trifuoroacetic acid, “Et2O” means diethylether, “DMSO” means dimethylsulfoxide, “SiO2” means silica, “XPhos Pd G3” means (2-dicyclohexylphosphino-2′,4′, 6′-triisopropyl-1, 1′-biphenyl)[2-(2′-amino-1,1′-biphenyl)]palladium(II) ethanesulfonate, “CDCl3” means deuterated chloroform, “MW” means microwave or molecular weight, “min” means minutes, “h” means hours, “rt” means room temperature, “quant” means quantitative, “n.t.” means not tested, “Cpd” means compound, “POCl3” means phosphorus(V) oxychloride.
For key intermediates, as well as some final compounds, the absolute configuration of chiral centers (indicated as R and/or S) were established via comparison with samples of known configuration, or the use of analytical techniques suitable for the determination of absolute configuration, such as VCD (vibrational cicular dichroism) or X-ray crystallography. When the absolute configuration at a chiral center is unknown, it is arbitrarily designated R*.
A solution of methyl trans-3-methoxyacrylate [34846-90-7] (20 mL, 1.08 g/mL, 186.02 mmol) in dry THF (245 mL) and 2,2,6,6-Tetramethylpiperidinylmagnesium chloride lithium chloride complex solution [898838-07-8] (265.75 mL, 0.77 M, 204.62 mmol) were pumped through a 10 mL coil at 40° C. (2.5 mL/min each line, 2.5 min residence time). The following out solution collected over a solution of copper cyanide [544-92-3] (18.33 g, 204.62 mmol) and lithium chloride [7447-41-8] (17.35 g, 409.25 mmol) in dry THF (200 mL) at 20° C. (water bath). The mixture was stirred for 20 min at RT. A solution of isobutyryl chloride [79-30-1] (23.32 mL, 223.23 mmol) in dry THF (90 mL) was added (drop funnel) at 20° C. and the mixture was stirred for 30 min at RT. Then sat. NaHCO3 in water (357 mL) and 8% aq. NH3 (438 mL) were added and the mixture was extracted with Et2O. The organic layer was separated, dried (MgSO4), filtered and concentrated in vacuo (minimum vacuum: 150 mbar) to afford a solution.
This solution was taken up with EtOH (288 mL) and hydrazinium hydroxide [7803-57-8] (55.64 mL, 744.09 mmol) was added. The reaction mixture was stirred at 120° C. for 1 h. The mixture was concentrated in vacuo and taken up with DCM. Then acidified with IM HCI (pH=2). The solid was filtered off through a celite pad and the organic layer separated, dried (MgSO4), filtered and the solvent evaporated. The crude product was purified by flash column chromatography (EtOAc in DCM 0/100 to 100/0). The desired fractions were collected and concentrated. The solid was washed with EtOH and dried to yield 6-isopropyl-5-methoxypyridazin-3(2H)-one 1A (16.37 g, 52%) as a white solid. 1H NMR (300 MHz, CDCl3) δ 1.19 (d, J=6.8 Hz, 6H), 3.13 (hept, J=6.8 Hz, 1H), 3.83 (s, 3H), 6.10 (s, 1H), 10.70 (s, 1H).
Ethyl bromoacetate [105-36-2] (10.5 mL, 92.80 mmol) was added to a stirred suspension of 6-isopropyl-5-methoxypyridazin-3(2H)-one 1A (14.32 g, 85.14 mmol) and Cs2CO3 [534-17-8] (41.61 g, 127.71 mmol) in ACN (116 mL) and DMF (55 mL) at RT. The reaction mixture was stirred in a metallic reactor at 120° C. (preheated oil bath) for 30 min. The crude was filtrated through celite and washed with EtOAc. The filtrate solvents were evaporated and the residue was purified by flash column chromatography (EtOAc in heptane 0/100 to 100/0). The desired fractions were collected and concentrated in vacuo to afford ethyl 2-(3-isopropyl-4-methoxy-6-oxopyridazin-1(6H)-yl) acetate 2A (20.82 g, 95%) as an oil that precipitates as a whitish solid upon standing.
1H NMR (300 MHz, CDCl3) δ 1.18 (d, J=6.8 Hz, 6H), 1.28 (t, J=7.1 Hz, 3H), 3.12 (hept, J=6.9 Hz, 1H), 3.82 (s, 3H), 4.23 (q, J=7.1 Hz, 2H), 4.80 (s, 2H), 6.12 (s, 1H)
Ethyl 2-(3-isopropyl-4-methoxy-6-oxopyridazin-1(6H)-yl) acetate 1B (19.2 g, 75.51 mmol) was put into several sealed vials (12×1600 mg) and purged and filled with nitrogen three times. Dry ACN (168 mL, 12×14 mL) was added and the solid was dissolved. Phosphoryl chloride (14.04 mL, 12×1.17 mL, 151.01 mmol) was added and the mixture was heated at 160° C. for 20 min under microwave irradiation. All the different reactions were combined and the excess of phosphoryl chloride was quenched with ice-water and the mixture was extracted with EtOAc. The organic layers were separated, combined, dried (MgSO4), filtered and the solvents evaporated in vacuo. The residue was purified by flash column chromatography (EtOAc in heptane 0/100 to 10/90). The desired fractions were collected and concentrated in vacuo to afford the ethyl 2-(4-chloro-3-isopropyl-6-oxopyridazin-1(6H)-yl) acetate 1C (14.95 g, 76%) as a clear yellow oil.
1H NMR (400 MHZ, CDCl3) δ 1.23 (d, J=6.8 Hz, 6H), 1.28 (t, J=7.1 Hz, 3H), 3.25 (hept, J=6.7 Hz, 1H), 4.24 (q, J=7.1 Hz, 2H), 4.83 (s, 2H), 7.01 (s, 1H).
A solution of LiOH [1310-65-2] (2.8 g, 116.92 mmol) in water (40 mL) was added to a stirred solution of 2-(4-chloro-3-isopropyl-6-oxopyridazin-1(6H)-yl) acetate 1C (6 g, 23.19 mmol) in 1,4-dioxane (80 mL). The mixture was stirred at RT for 2h. The mixture was treated with 2 N HCl in water to reach pH 2 and then extracted with EtOAc (50 mL×4) and with a mixture THF/EtOAc (3/7, 60 mL). The combined organic layers were dried (Na2SO4), filtered and the solvents evaporated in vacuo to yield 2-(4-chloro-3-isopropyl-6-oxopyridazin-1(6H)-yl) acetic acid 1D (5.32 g, quant.) as a yellow solid. LCMS (Rt: 0.47, Area %: 100, MW: 230.0, BPM1: 231.05, Method 6)
1H NMR (500 MHZ, DMSO-d6) 1.18 (d, J=6.71 Hz, 6 H) 3.15-3.26 (m, 1 H) 4.72 (s, 2 H) 7.31 (s, 1 H): 1H exchanged
DIPEA [7087-68-5] (11.2 mL, 64.99 mmol) was added to a stirred solution of 2-(4-chloro-3-isopropyl-6-oxopyridazin-1(6H)-yl) acetic acid 1D (2.8 g, 12.14 mmol), [1,2,4]triazolo[4,3-A]pyridin-7-amine [1082448-58-5] (1.79 g, 13.35 mmol) and HATU [148893-10-1] (5.15 g, 13.55 mmol) in DMF (56 mL). The mixture was stirred at RT for 2.5 h. The mixture was diluted with sat. NaHCO3 in water and extracted with EtOAc (100 mL×4) and then with a mixture EtOAc/THF (7/3, 70 mL×2). The combined organic layers were dried (Na2SO4), filtered and the solvents evaporated in vacuo to yield a beige solid.
This solid was triturated with ACN, filtered and washed with additional ACN to yield N-([1,2,4] triazolo[4,3-a] pyridin-6-yl)-2-(4-chloro-3-isopropyl-6-oxopyridazin-1(6H)-yl) acetamide 1E (3.48 g, 83%) as an off white solid.
The filtrate was evaporated in vacuo and purified by flash column chromatography (silica: MeOH in DCM 0/100 to 10/90). The desired fractions were collected and concentrated in vacuo to yield additional N-([1,2,4] triazolo[4,3-a] pyridin-6-yl)-2-(4-chloro-3-isopropyl-6-oxopyridazin-1(6H)-yl) acetamide 1E (334 mg, 8%) as a beige solid.
1H NMR (400 MHZ, DMSO-d6) 8 ppm 1.20 (d, J=6.94 Hz, 6 H) 3.18-3.29 (m, 1 H) 4.92 (s, 2 H) 7.29 (dd, J=9.71, 1.85 Hz, 1 H) 7.34 (s, 1 H) 7.79 (d, J=9.71 Hz, 1 H) 9.20 (dd, J=1.62, 0.92 Hz, 1 H) 9.23 (d, J=0.69 Hz, 1 H) 10.39-10.83 (m, 1 H).
A solution of 1-(2-oxabicyclo[2.1.1]hexan-1-ylmethyl)pyrazol-4-amine (555 mg, 3.1 mmol), sodium trifluoromethanesulfinate [2926-29-6] (725 mg, 4.65 mmol), ammonium persulfate [7727-54-0] (1.06 g, 4.65 mmol) and (Ir[dF(CF3)ppy]2(dtbpy))PF6 [870987-63-6] (52.12 mg, 0.046 mmol) in DMSO (28 mL) was prepared in a thin tube. The reaction mixture was reacted in a Vapourtec Photoreactor UV-150 irradiating with blue LED 450nm with a retention time of 30 min, coil 10 mL (flow rate 0.333 mL/min). The reaction was diluted with EtOAc (50 mL) and water (50 mL). The layers were separated and the aqueous layer was further extracted with EtOAc (2×50 mL). The combined organic layers were dried (MgSO4), filtered and concentrated in vacuo. The crude product was purified by flash column chromatography (silica, EtOAc in DCM 0/100 to 100/0). Desired fractions were collected and the solvent evaporated to yield 1-(2-oxabicyclo[2.1.1 ]hexan-1-ylmethyl)-5-(trifluoromethyl)pyrazol-4-amine 1F (144 mg, 19%) as a dark brown oil, that was used in the next step without further purification.
Aniline [62-53-3] (10 μL, 0.11 mmol) was added to a stirred suspension of 1E (30 mg, 0.087 mmol), Pd2(dba)3 [51364-51-3] (4 mg, 4.4 μmol), Xantphos [161265-03-8] (5.01 mg, 8.7 μmol) , and Na2CO3 (20 mg, 0.19 mmol) in a mixture of 1,4-dioxane (8 mL) and DMF (0.05 mL) under N2. The mixture was stirred at 175° C. for 20 min under microwave irradiation. The crude mixture was filtered through celite and washed with EtOAc. The solvent was concentrated in vacuo and the crude product was purified by RP HPLC (stationary phase: C18 XBridge 30×100 mm 5 um), mobile phase: gradient from 85% NH4HCO3 0.25% solution in water, 15% ACN to 55% NH4HCO3 0.25% solution in water, 45% ACN), yielding 2-(4-anilino-3-isopropyl-6-oxo-pyridazin-1-yl)-N-([1,2,4]triazolo[4,3-a]pyridin-6-yl)acetamide (Final compound 1) (10.7 mg, 31%) as an off white solid.
Additional analogues were accessed using similar reaction conditions, using the appropriate reagent.
[2137995-69-6]
[753-90-2]
DIPEA [7087-68-5] (80 μL, 0.5 mmol) was added to a stirred mixture of 1E (67.6 mg, 0.16 mmol) and pyrrolidine [123-75-1] (19.4 μL, 0.2 mmol) in ACN (1 mL). The mixture was stirred at 150° C. for 15 min under microwave irradiation. Then sat. Na2CO3 (1.5 mL) was added and the mixture was extracted with DCM. The organic layer was separated, dried (Na2SO4), filtered and the solvent evaporated in vacuo. The crude product was triturated with EtOAc to yield 2-(3-isopropyl-6-oxo-4-pyrrolidin-1-yl-pyridazin-1-yl)-N-([1,2,4]triazolo[4,3-a]pyridin-6-yl)acetamide (Final compound 5) (34 mg, 57%) as an off-white solid.
Additional analogues were accessed using similar reaction conditions, using the appropriate reagent.
[110-91-8]
[1003-03-8]]
[63126-47-6]
[136725-53-6]
[136725-55-8]
[23356-96-9]
[21987-29-1]
[163457-23-6]
[285-59-6]
[63126-47-6]
[120099-61-8]
[120099-60-7]
[59335-84-1]
[1003-28-7]
[51503-10-7]
[27514-07-4]
[13220-27-4]]
[935669-28-6]
[148644-09-1]
[106404-52-8]
[256931-54-1]
[132958-72-6]
[132883-44-4]
DIPEA [7087-68-5] (56 μL, 0.35 mmol) was added to a stirred suspension of 1E (40 mg, 0.12 mmol) and cyclopropanemethylamine [2516-47-4] (16 μL, 0.18 mmol) in ACN (1.5 mL) and DMF (0.5 mL). The mixture was stirred at 150° C. for 15 min under microwave irradiation. Then, additional cyclopropanemethylamine [2516-47-4] (50 μL, 0.58 mmol) and DMF (0.5 mL) were added and the mixture was stirred at 150° C. for 20 min under microwave irradiation and at 100° C. for 18 h. Then, the mixture was diluted with 10% sol. aq. NH4HCO3 and DCM and the organic phase was separated by an Isolute® phase separator, The organic layer was evaporated in vacuo and the crude product was purified by RP HPLC (stationary phase: C18 XBridge 30×100 mm 5 μm), mobile phase: gradient from 70% NH4HCO3 0.25% solution in water, 30% ACN to 35% NH4HCO3 3 0.25% solution in water, 65% ACN), yielding 2-[4-(cyclopropylmethylamino)-3-isopropyl-6-oxo-pyridazin-1-yl]-N-([1,2,4]triazolo[4,3-a]pyridin-6-yl)acetamide (Final compound 29) (13.2 mg, 30%) and 2-[4-(dimethylamino)-3-isopropyl-6-oxo-pyridazin-1-yl]-N-([1,2,4]triazolo[4,3-a]pyridin-6-yl)acetamide (Final compound 30) (7.4 mg, 18%) as off-white solids.
A vial was charged with 1E (30 mg, 86.51 μmol), NMM (52.503 mg, 0.519 mmol) and 3,3-dimethylcyclobutan-1-amine [123788-48-7] (34.32 mg, 0.35 mmol) in ACN (0.1 mL) and DMI (0.4 mL). The mixture was heated to 120° C. for 72 hours. Contents were cooled to ambient and solvents and volatiles distilled to dryness. The residue was dissolved in MeOH/DMSO/ammonium bicarbonate (2.5 mL), filtered and purified by RP HPLC to obtain N-([1,2,4]triazolo[4,3-a]pyridin-6-yl)-2-(4-((3,3-dimethylcyclobutyl)amino)-3-isopropyl-6-oxopyridazin-1(6H)-yl)acetamide (Final compound 31) (5.46 mg, 15% yield).
Additional analogues were accessed using similar reaction conditions, using the appropriate reagent.
[1375472-05-1]
[1414958-74-9]
[1346808-65-8]
[1785540-77-3]
[3970-68]
6-Cyclopropylpyridin-2-amine [857292-66-1] (17.41 mg, 0.13 mmol) was added to a vial in the glove box and the scaffold 1E was added from a stock solution in degassed dioxane (0.5 mL of 0.17 M stock solution, 0.087 mmol, 1 eq.). Next Xantphos Pd G4 [1621274-19-8] was added from a stock solution in dioxane (0.1 mL of 0.087 M stock, 0.15 eq.). Cesium carbonate (112.748 mg, 0.346 mmol) was added as a solid. The vial was heated at 120 ° C. for 16 h on a tumble stirrer. After cooling to room temperature, the reaction mixture was diluted with ethyl acetate (2 mL) and DMT scavenger (5 eq.) was added and stirred for 1 h. The scavenger was removed by filtration, and the solvent removed under vacuum. The dry material was re-dissolved in DMSO (3 mL) and a purification was performed via Prep HPLC (Stationary phase: RP XBridge Prep C18 OBD-10 μm, 30×150 mm, Mobile phase: 0.25% NH4HCO3 3 solution in water, CH3CN or MeOH) to obtain N-([1,2,4]triazolo[4,3-a]pyridin-6-yl)-2-(4-((6-cyclopropylpyridin-2-yl)amino)-3-isopropyl-6-oxopyridazin-1(6H)-yl)acetamide (Final compound 38) (0.63 mg, 2% yield).
Additional analogues were accessed using similar reaction conditions, using the appropriate reagent.
[1152519-69-1]
Example A6
Synthesis of N-([1,2,4]triazolo[4,3-a]pyridin-6-yl)-2-(4-((1-(2,2-difluorocyclopropyl)ethyl)amino)-3-isopropyl-6-oxopyridazin-1(6H)-yl)acetamide (Final compound 40)
1-(2,2-Difluorocyclopropyl)ethan-1-amine [1909336-12-4] (15.72 mg, 0.17 mmol, 2.0 eq.) were added to a vial into the glove box and the scaffold 1E (30 mg, 0.0865 mmol) was added from a stock solution in degassed dioxane (0.5 mL of 0.17 M stock solution, 0.087 mmol, 1 eq.). Next, RuPhos Pd G4 [1599466-85-9] was added from a stock solution in dioxane (0.1 mL of 0.087 M stock, 0.10 eq.). NaOtBu (33.257 mg, 0.346 mmol, 4 eq.) was added as a solid. The vial was heated at 120° C. for 16 h on a tumble stirrer. After cooling to room temperature, the reaction mixture was diluted with ethyl acetate (2 mL) and DMT scavenger (5 eq.) was added and stirred for 1 h. The scavenger was removed by filtration, and the solvent removed under vacuum. The dry material was re-dissolved in DMSO (3 mL) and a purification was performed via Prep HPLC
(Stationary phase: RP XBridge Prep C18 OBD-10 μm, 30×150mm, Mobile phase: 0.25% NH4HCO3 3 solution in water, CH3CN or MeOH) to obtain N-([1,2,4]triazolo[4,3-a]pyridin-6-yl)-2-(4-((1-(2,2-difluorocyclopropyl)ethyl)amino)-3-isopropyl-6-oxopyridazin-1(6H)-yl)acetamide (Final compound 40) (0.88 mg, 2% yield).
Benzo[d]isoxazol-5-amine [239097-74-6] (23.21 mg, 0.17 mmol, 2.0 eq.) was added to a vial. The scaffold 1E was added from a stock solution in dry DMSO (0.5 mL of 0.17 M stock solution, 0.087 mmol, 1 eq.). Next Pd-PEPPSI-IPent (6.866 mg, 0.00865 mmol, 0.1 eq.) was added from a stock solution in DMSO (0.1 mL of 0.087 M stock, 0.10 eq.). Potassium phosphate tribasic (73.454 mg, 0.346 mmol, 4 eq.) was added as a solid. The vial was heated at 120° C. for 64 h on a tumble stirrer. After cooling to room temperature, the reactions mixtures were diluted with ethyl acetate (2 mL) and DMT scavenger (5 eq.) was added and stirred for 1 h. The scavenger was removed by filtration, and the solvent removed under vacuum. The dry material was re-dissolved in DMSO (3 mL) and a purification was performed via Prep HPLC (Stationary phase: RP XBridge Prep C18 OBD-10 μm, 30×150 mm, Mobile phase: 0.25% NH4HCO3 3 solution in water, CH3CN or MeOH) to obtain N-([1,2,4]triazolo[4,3-a]pyridin-6-yl)-2-(4-(benzo[d]isoxazol-5-ylamino)-3-isopropyl-6-oxopyridazin-1(6H)-yl)acetamide (Final compound 41) (14.4 mg, 37% yield).
Additional analogues were accessed using similar reaction conditions, using the appropriate reagent.
An 8 mL vial was charged with 1E (30 mg, 86.51 μmol), DIPEA [7087-68-5] (42 uL, 0.8 g/mL, 0.26 mmol) and a collection of piperidine [110-89-4 ] (11.05 mg, 1.5 eq, 0.13 mmol) in ACN (2 mL) as solvent. The mixture was warmed to 120 ° C.for 72 hours. Contents were cooled to ambient and solvents and volatiles distilled to dryness. The corresponding residue was dissolved in MeOH/DMSO/ammonium bicarbonate (2.5 mL), filtered and purified by RP HPLC (Stationary phase: C18 XBridge 30×100 mm 5 um), Mobile phase: NH4HCO3 0.25% solution in Water and CH3CN, yielding N-([1,2,4]triazolo[4,3-a]pyridin-6-yl)-2-(3-isopropyl-6-oxo-4-(piperidin-1-yl)pyridazin-1(6H)-yl)acetamide (Final compound 45) (24.6 mg, yield 72%).
Additional analogues were accessed using similar reaction conditions, using the appropriate reagent.
[78-81-9]
[625-43-4]
[109-01-3]
[665-04-3]
[174-78-7]
[186597-29-5]
[176-12-5]
[1427367-47-2]
[109-05-7]
[17305-22-5]
[35794-11-7]
[1316223-46-7]
[1215071-12-7]
[253603-61-1]
[116574-75-5]
[1263132-31-5]
[710943-30-9]
[524674-44-0]
[630121-89-0]
[150281-47-3]
[765-30-0]
[5163-20-2]
This is depicted in the following table:
1H NMR (400 MHz, DMSO-d6) δ ppm 1.21 (d, J = 6.47 Hz, 6 H) 2.52
1H NMR (400 MHz, DMSO-d6) δ ppm 1.18 (d, J = 6.71 Hz, 6 H) 1.32
1H NMR (400 MHz, DMSO-d6) δ ppm 1.11-1.31 (m, 6 H) 1.36 (dd,
1H NMR (400 MHz, DMSO-d6) δ ppm 1.14 (d, J = 6.70 Hz, 6 H) 3.09
1H NMR (400 MHz, DMSO-d6) δ ppm 1.15 (d, J = 6.47 Hz, 6 H)
1H NMR (400 MHz, DMSO-d6) δ ppm 1.17 (d, J = 6.6 Hz, 6 H) 2.90-2.97
1H NMR (400 MHz, DMSO-d6) δ ppm 1.18 (d, J = 6.6 Hz, 6 H) 2.09-2.21
1H NMR (400 MHz, DMSO-d6) δ ppm 1.05-1.09 (m, 6 H) 1.22 (d,
1H NMR (400 MHz, DMSO-d6) δ ppm 1.13 (d, J = 6.6 Hz, 6 H) 2.94
1H NMR (400 MHz, DMSO-d6) δ ppm 0.20-0.28 (m, 2 H) 0.43-0.52
1H NMR (400 MHz, DMSO-d6) δ ppm 1.16 (d, J = 6.70 Hz, 6 H) 2.75
1H NMR (500 MHz, DMSO-d6) δ ppm 1.13 (d, J = 6.56 Hz, 6 H) 1.80
1H NMR (500 MHz, DMSO-d6) δ ppm 1.06 (d, J = 6.71 Hz, 3 H) 1.11
1H NMR (500 MHz, DMSO-d6) δ ppm 0.92 (d, J = 6.56 Hz, 3 H) 1.00-1.11
1H NMR (400 MHz, DMSO-d6) δ ppm 0.71 (q, J = 11.87 Hz, 1 H)
1H NMR (400 MHz, DMSO-d6) δ ppm 1.17 (dd, J = 6.47, 3.93 Hz, 6 H)
A compound of the invention (for instance, a compound of the examples) is brought into association with a pharmaceutically acceptable carrier, thereby providing a pharmaceutical composition comprising such active compound. A therapeutically effective amount of a compound of the invention (e.g. a compound of the examples) is intimately mixed with a pharmaceutically acceptable carrier, in a process for preparing a pharmaceutical composition.
The activity of a compound according to the present invention can be assessed by in vitro methods. A compound the invention exhibits valuable pharmacological properties, e.g. properties susceptible to inhibit NLRP3 activity, for instance as indicated the following test, and are therefore indicated for therapy related to NLRP3 inflammasome activity.
Peripheral venous blood was collected from healthy individuals and human peripheral blood mononuclear cells (PBMCs) were isolated from blood by Ficoll-Histopaque (Sigma-Aldrich, A0561) density gradient centrifugation. After isolation, PBMCs were stored in liquid nitrogen for later use. Upon thawing, PBMC cell viability was determined in growth medium (RPMI media supplemented with 10% fetal bovine serum, 1% Pen-Strep and 1% L-glutamine). Compounds were spotted in a 1:3 serial dilution in DMSO and diluted to the final concentration in 30 μl medium in 96 well plates (Falcon, 353072). PBMCs were added at a density of 7.5×104 cells per well and incubated for 30 min in a 5% CO2 incubator at 37 ° C. LPS stimulation was performed by addition of 100 ng/ml LPS (final concentration, Invivogen, tlrl-smlps) for 6 hrs followed by collection of cellular supernatant and the analysis of IL-1β(μM) and TNF cytokines levels (uμ) via MSD technology according to manufacturers' guidelines (MSD, K151A0H).
IC50 and AC50 values (for IL-1β) and EC50 and AC50 values (TNF) were obtained on compounds of the invention/examples, and the AC50 values are depicted in the following table:
One or more compound(s) of the invention (including compounds of the final examples) is/are tested in a number of other methods to evaluate, amongst other properties, permeability, stability (including metabolic stability and blood stability) and solubility.
The in vitro passive permeability and the ability to be a transported substrate of
P-glycoprotein (P-gp) is tested using MDCK cells stably transduced with MDR1 (this may be performed at a commercial organisation offering ADME, PK services, e.g. Cyprotex). Permeability experiments are conducted in duplicate at a single concentration (5 uM) in a transwell system with an incubation of 120 min. The apical to basolateral (AtoB) transport in the presence and absence of the P-gp inhibitor GF120918 and the basolateral to apical (BtoA) transport in the absence of the P-gp inhibitor is measured and permeation rates (Apparent Permeability) of the test compounds (Papp×10−6 cm/sec) are calculated.
The metabolic stability of a test compound is tested (this may be performed at a commercial organization offering ADME, PK services, e.g. Cyprotex) by using liver microsomes (0.5 mg/ml protein) from human and preclinical species incubated up to 60 minutes at 37° C. with 1 μM test compound.
The in vitro metabolic half-life (t1/2) is calculated using the slope of the log-linear regression from the percentage parent compound remaining versus time relationship (κ),
t1/2=−In(2)/κ.
The in vitro intrinsic clearance (Clint) (ml/min/mg microsomal protein) is calculated using the following formula:
Where: Vinc=incubation volume,
The metabolic stability of a test compound is tested using liver hepatocytes (1 milj cells) from human and preclinical species incubated up to 120 minutes at 37° C. with 1 μM test compound.
The in vitro metabolic half-life (t1/2) is calculated using the slope of the log-linear regression from the percentage parent compound remaining versus time relationship (κ), t1/2=−In(2)/κ.
The in vitro intrinsic clearance (Clint) (μl/min/million cells) is calculated using the following formula:
Where: Vinc=incubation volume,
The test/assay is run in triplicate and is semi-automated using the Tecan Fluent for all liquid handling with the following general steps:
The LC conditions are:
The compound of the invention/examples is spiked at a certain concentration in plasma or blood from the agreed preclinical species; then after incubating to predetermined times and conditions (37° C., 0° C. (ice) or room temperature) the concentration of the test compound in the blood or plasma matrix with LCMS/MS can then be determined.
| Number | Date | Country | Kind |
|---|---|---|---|
| 21160669.4 | Mar 2021 | EP | regional |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/EP2022/055433 | 3/3/2022 | WO |
