cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-1,3-diazaspiro[4.5]decane-2-one derivative

Abstract

The present invention relates to cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-1,3-diazaspiro[4.5]-decan-2-one derivatives, their preparation and use in medicine, in particular, but not limited to, use in various neurological disorders including pain, neurodegenerative diseases, neuroinflammatory diseases, neuropsychiatric disorders and substance abuse / dependence.

Description

[Technical Field]

[0001] We claim priority to European Patent Application No. 23169716.0, filed on April 25, 2023, and to U.S. Provisional Patent Application No. 63 / 461,659, filed on April 25, 2023.

[0002] The present invention relates to cis-8-(3,5-difluorophenyl)-8-(alkylamino)-1,3-diazaspiro[4.5]-decan-2-one derivatives, their preparation and use in medicine, in particular, but not limited to, use in various neurological disorders including pain, neurodegenerative diseases, neuroinflammatory diseases, neuropsychiatric disorders and substance abuse / dependence.

[0003] Opioid receptors are a group of Gi / o protein-coupled receptors widely distributed in the human body. Currently, opioid receptors are subdivided into four main classes: the three classical opioid receptors—mu-opioid (MOP) receptors, kappa-opioid (KOP) receptors, and delta-opioid (DOP) receptors—and the opioid receptor-like (ORL-1) receptor, which was recently discovered based on its high homology to the aforementioned classical opioid receptors. After the identification of the endogenous ligand of the ORL-1 receptor, a highly basic 17-amino acid peptide known as nociceptin / orphanin FQ, isolated from tissue extracts in 1995, the ORL-1 receptor was renamed the "nociceptin opioid peptide receptor" and abbreviated as the "NOP receptor."

[0004] Classical opioid receptors (MOP, KOP, and DOP), as well as NOP receptors, are widely distributed and expressed throughout the human body, including the brain, spinal cord, peripheral sensory neurons, and intestinal tract, and the patterns of distribution differ among various receptor classes.

[0005] Nociceptin acts at the molecular and cellular levels, much like opioids. However, the pharmacological effects of nociceptin can sometimes differ from, and even counteract, those of opioids. Activation of NOP receptors leads to complex pharmacology in pain modulation, resulting in either pro-algesic or anti-algesic activity depending on the administration route, pain model, and species involved. Furthermore, the NOP receptor system is upregulated under chronic pain conditions. Systemic administration of selective NOP receptor agonists has been found to exert potent and effective analgesia in non-human primate models of acute inflammatory pain without side effects. Activation of NOP receptors has been demonstrated to inhibit, rather than enhance, the effects of opioid-mediated reward in rodents and non-human primates (overview: Schroeder et al, Br J Pharmacol 2014;171(16):3777-3800, and references in the relevant literature).

[0006] In addition to the involvement of NOP receptors in pain perception, results from preclinical studies suggest that NOP receptor agonists may be particularly useful in the treatment of neuropsychiatric disorders (Witkin et al, Pharmacology & Therapeutics, 141 (2014) 283-299; Jenck et al., Proc. Natl. Acad. Sci. USA 94, 1997, 14854-14858).

[0007] Potent opioids acting at the MOP receptor site are widely used to treat moderate to severe acute and chronic pain. However, the therapeutic range of potent opioids is limited by serious side effects such as nausea and vomiting, constipation, dizziness, somnolence, respiratory depression, physical dependence, and abuse. Furthermore, MOP receptor agonists are known to simply be less effective in chronic and neuropathic pain conditions.

[0008] Alternatively, the delta-opioid receptor (DOP) has been a target of interest for many years as a potential treatment for pain, as well as anxiety and depression. Selective DOP ligands have generally failed due to a lack of efficacy, but significant safety issues such as seizures, effects on spontaneous / coordinated movement, tolerance, headache, and gastrointestinal discomfort have been reported in preclinical and clinical trials (Broom et al, 2002; Hudzik et al, 2014; Spahn & Stein, 2017). These safety-related effects have been shown to be distinguishable by selective DOP ligands, and new evidence suggesting diverse downstream signaling pathways, receptor phosphorylation, receptor transport, and selectivity adds complexity to understanding the biological and pharmacological responses of DOP ligands (Broon et al, 2002; Pradhan et al, 2012; Mann et al, 2020; Quirion et al, 2020).

[0009] Some of the potent opioid side effects mentioned above are known to be mediated by the activation of classical opioid receptors in the central nervous system. Furthermore, peripheral opioid receptors, when activated, may inhibit the transmission of pain signals, as shown in both clinical and animal studies (Gupta et al., 2001; Kalso et al., 2002; Stein et al., 2003; Zollner et al., 2008).

[0010] Therefore, one approach to avoid CNS-mediated adverse effects after systemic administration is to provide peripherally restricted opioid receptor ligands that do not readily cross the blood-brain barrier and are therefore not distributed much to the central nervous system (see, for example, WO2015 / 192039). Such peripherally acting compounds may provide a combination of effective analgesia and limited side effects.

[0011] Another approach is to provide compounds that interact with both NOP receptors and MOP receptors. Such compounds are described, for example, in WO2004 / 043967, WO2012 / 013343, and WO2009 / 118168.

[0012] A further approach would be to provide multi-opioid receptor analgesics that modulate two or more opioid receptor subtypes to result in additive or synergistic analgesia and / or reduced side effects such as abuse tendency or tolerance.

[0013] On the one hand, it is desirable to provide an analgesic that selectively acts on the NOP receptor system and does not act so significantly on the classical opioid receptor system, while on the other hand, it is desirable to distinguish between central nervous system activity and peripheral nervous system activity.

[0014] 8-(alkylamino)-1,3-diazaspiro[4.5]decane-2-one derivatives are known from, for example, WO 2017 121646, WO 2017 121647, WO 2017 121648, WO 2017 121649, WO 2017 121650, and WO 2019 012037.

[0015] There is a need for pharmaceuticals that are effective in treating pain and offer advantages over conventional compounds. Whenever possible, such pharmaceuticals should contain low doses of active ingredients to ensure adequate pain relief without causing unacceptable adverse events associated with treatment.

[0016] The object of the present invention is to provide a pharmacologically active compound, preferably an analgesic, that has advantages over the prior art.

[0017] This objective was achieved by the subject matter of the patent claims.

[0018] The present invention relates to the compound described in general formula (I), [ka] During the ceremony, A1 represents N or CR1, A2 represents N or C, A3 represents N or CH, A4 represents N or CH, A5 represents N or C, (However, one or two of A1, A2, A3, A4, and A5 represent N, and the remaining three or four of A1, A2, A3, A4, and A5 do not represent N), R1 represents -H, -CH3, or =O. R2 is -C1-C6-alkyl (linear or branched, saturated or unsaturated, unsubstituted, or substituted with one, two, three, or four substituents independently selected from -F and -OH); -C3-C7-heterocycloalkyl (saturated or unsaturated, unsubstituted, or substituted with one, two, three, or four substituents independently selected from -F, -OH, and -CH3); -S(O)2-C1-C4-alkyl (linear, branched, or cyclic, unsubstituted, or substituted with one, two, or three substituents -F); -O-C1-C6-alkyl (linear or branched, saturated or unsaturated, unsubstituted, or substituted with one, two, three, or four substituents independently selected from -F, -OH, and -OCH3); -O-C3-C8-cycloalkyl (saturated or unsaturated, unsubstituted, or substituted with one, two, three, or four substituents independently selected from -F, -OH, and -CH3); or -O-C3-C7-heterocycloalkyl (saturated or unsaturated, unsubstituted, or substituted with one, two, three, or four substituents independently selected from -F and -OH); Alternatively, R1 and R2 together with A1 and A2 form a ring, meaning -CH=CN-NH-, R3 represents -H or -CH3, and Both R4 and R5 represent -H, or together they mean =O. or relating to physiologically acceptable salts thereof.

[0019] The compounds according to the present invention all have a novel structural motif not known from the prior art (for example, ). This structural motif is characterized by a 3,5-difluorophenyl moiety and a methyl group at A5.

[0020] As demonstrated by comparative experimental data, the novel structural motif has the effect of consistently achieving higher selectivity with respect to DOP.

[0021] According to the present invention, unless otherwise specified, the "-C1-C6-alkyl" and any other alkyl residues may be saturated linear or branched. Examples of linear saturated alkyls include methyl, ethyl, n-propyl, n-butyl, n-pentyl, and n-hexyl. Examples of branched saturated alkyls include, but are not limited to, isopropyl, sec-butyl, and tert-butyl.

[0022] In the present invention, "C3-C8-cycloalkyl" means a non-aromatic, monocyclic or bicyclic moiety that contains 3 to 8 ring carbon atoms but does not contain heteroatoms within the ring. Preferred saturated C3-C8-cycloalkyls in the present invention include, but are not limited to, cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, and cyclooctane. Preferred unsaturated C3-C8-cycloalkyls in the present invention include, but are not limited to, cyclopropene, cyclobutene, cyclopentene, cyclopentadiene, cyclohexene, 1,3-cyclohexadiene, and 1,4-cyclohexadiene.

[0023] In the present invention, "C3-C7-heterocycloalkyl" means a non-aromatic, monocyclic, or bicyclic moiety containing 4 to 8 ring atoms, where each ring independently contains 1, 2, 3, 4, or more heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur, the sulfur may be oxidized (S(=O) or S(=O)2), the remaining ring atoms are carbon atoms, and the bicyclic system may share one or more common heteroatoms. Preferred saturated C3-C7-heterocycloalkyls according to the present invention include, but are not limited to, azetidine, pyrrolidine, oxetane, and tetrahydrofuran.

[0024] In a preferred embodiment, both R4 and R5 represent -H.

[0025] In a preferred embodiment, one of A1, A2, A3, A4, and A5 represents N, and the remaining four of A1, A2, A3, A4, and A5 do not represent N.

[0026] In other preferred embodiments, two of A1, A2, A3, A4, and A5 represent N, and the remaining three of A1, A2, A3, A4, and A5 do not represent N.

[0027] In a preferred embodiment, according to general formula (II-A), A1 represents CR1; A2 represents C; A3 represents CH; A4 represents N; and A5 represents C: [ka]

[0028] When R3 represents -CH3 and R4 and R5 each represent -H, this structural motif is realized by Examples EX-I-01, EX-I-03, EX-I-04, EX-I-05, EX-I-07, EX-I-09, EX-I-10, EX-I-11, EX-I-24, and EX-I-26.

[0029] When R3, R4, and R5 each represent -H, this structural motif is realized by Examples EX-I-06 and EX-I-25.

[0030] If R3 represents -CH3 and R4 and R5 together mean =O, this structural motif is realized by Example EX-I-28.

[0031] In a preferred embodiment, according to general formula (II-B), A1 represents N; A2 represents C; A3 represents CH; A4 represents CH; and A5 represents C: [ka]

[0032] This structural motif is realized by Examples EX-I-12, EX-I-13, EX-I-14, and EX-I-17.

[0033] In a preferred embodiment, according to general formula (II-C), A1 represents CR1; A2 represents C; A3 represents N; A4 represents N; and A5 represents C: [ka]

[0034] This structural motif is realized in Examples EX-I-08 and EX-I-27.

[0035] In a preferred embodiment, according to general formula (II-D), A1 represents N; A2 represents C; A3 represents N; A4 represents CH; and A5 represents C: [ka]

[0036] This structural motif is realized by Examples EX-I-02, EX-I-15, EX-I-16, EX-I-18, EX-I-21, and EX-I-22.

[0037] In a preferred embodiment, according to general formula (II-E), A1 represents CR1, R1 means = O; A2 represents N; A3 represents CH; A4 represents CH; and A5 represents C: [ka]

[0038] This structural motif is realized by Example EX-I-19.

[0039] In a preferred embodiment, according to general formula (II-F), A1 represents N; A2 represents C; A3 represents CH; A4 represents N; and A5 represents C: [ka]

[0040] This structural motif is realized by Example EX-I-20.

[0041] In a preferred embodiment, according to general formula (II-G), A1 represents CR1, R1 means = O; A2 represents C; A3 represents CH; A4 represents CH; and A5 represents N: [ka]

[0042] This structural motif is realized by Example EX-I-23.

[0043] In a preferred embodiment, R1 (if present) represents -H or -CH3.

[0044] In preferred embodiments, R2 is a -C1-C3-alkyl (linear or branched, saturated, unsaturated, or substituted with one, two, three, or four substituents independently selected from -F and -OH; preferably, representing -CH3, -CF3, -C(CH3)2OH, or -C(CH3)(OH)CF3).

[0045] In preferred embodiments, R2 represents a -C4-heterocycloalkyl (saturated, unsubstituted, or substituted with one, two, or three substituents independently selected from -F, -OH, and -OCH3; preferably 3-fluorooxetane-3-yl).

[0046] In preferred embodiments, R2 represents a -S(O)2-C1-C2-alkyl (linear, unsubstituted), preferably -S(O)2CH3 or -S(O)2CH2CH3.

[0047] In a preferred embodiment, R2 represents an -O-C1-C2-alkyl (saturated, unsubstituted, or substituted with one, two, or three substituents independently selected from -F, -OH, and -OCH3; preferably -OCH3, -O-CH2CH2-OCH3, -OCHF2, -OCH2CHF2, -OCF2CH3, or -OCF3).

[0048] In preferred embodiments, R2 represents -O-C3-C4-cycloalkyl (saturated, unsubstituted, or substituted with one or two substituents independently selected from -OH and -CH3; preferably -O-cyclopropyl or -O-(3-hydroxy-3-methyl)-cyclobutyl (i.e., 3-hydroxy-3-methylcyclobutoxy)).

[0049] In a preferred embodiment, R2 represents an -O-C4-heterocycloalkyl (saturated, unsubstituted; preferably -O-oxetan-3-yl).

[0050] In a preferred embodiment, R2 represents -CH3 according to general formula (III-A): [ka]

[0051] This structural motif is realized in Examples EX-I-13 and EX-I-19.

[0052] In a preferred embodiment, R2 represents -CF3 according to general formula (III-B): [ka]

[0053] This structural motif is realized by Example EX-I-12.

[0054] In a preferred embodiment, R2 represents -C(CH3)2OH according to the general formula (III-C): [ka]

[0055] R3 represents -CH3, and this structural motif is realized by Examples EX-I-03, EX-I-20, and EX-I-22. When R3 represents -H, this structural motif is realized by Example EX-I-06.

[0056] In a preferred embodiment, R2 represents -S(O)2CH3 according to general formula (III-D): [ka]

[0057] This structural motif is realized in Examples EX-I-10 and EX-I-17.

[0058] In a preferred embodiment, R2 represents -S(O)2CH2CH3 according to the general formula (III-E): [ka]

[0059] This structural motif is realized by Example EX-I-11.

[0060] In a preferred embodiment, R2 represents -OCH3 according to general formula (III-F): [ka]

[0061] This structural motif is realized by Example EX-I-02.

[0062] In a preferred embodiment, R2 represents -OCHF2 according to general formula (III-G): [ka]

[0063] When R4 and R5 each represent -H, this structural motif is realized by Example EX-I-04.

[0064] When R4 and R5 together mean = O, this structural motif is realized by Example EX-I-28.

[0065] In a preferred embodiment, R2 represents -OCF3 according to the general formula (III-H): [ka]

[0066] This structural motif is realized by Examples EX-I-07, EX-I-08, and EX-I-14.

[0067] In a preferred embodiment, R2 represents -OCH2CH2OCH3 according to general formula (III-I): [ka]

[0068] This structural motif is realized by Example EX-I-16.

[0069] In a preferred embodiment, R2 represents -O-cyclopropyl according to general formula (III-J): [ka]

[0070] This structural motif is realized by Examples EX-I-01, EX-I-18, and EX-I-25.

[0071] In a preferred embodiment, R2 represents 3-hydroxy-3-methylcyclobutoxy according to the general formula (III-K): [ka]

[0072] This structural motif is realized by Example EX-I-09.

[0073] In a preferred embodiment, R2 represents oxetane-3-yloxy according to the general formula (III-L): [ka]

[0074] This structural motif is realized by Example EX-I-15.

[0075] In a preferred embodiment, R1 and R2 together with A1 and A2 form a ring, which according to the general formula (III-M) means -CH=CN-NH-: [ka]

[0076] This structural motif is realized by Example EX-I-05.

[0077] In a preferred embodiment, R2 represents 3-fluorooxetane-3-yl according to the general formula (III-N): [ka]

[0078] This structural motif is realized by Example EX-I-21.

[0079] In a preferred embodiment, R2 represents -OCHF2 according to the general formula (III-O): [ka]

[0080] This structural motif is realized by Example EX-I-23.

[0081] In a preferred embodiment, R2 represents -C(CH3)(OH)CF3 according to the general formula (III-P): [ka]

[0082] This structural motif is realized by Example EX-I-24. Since R2 is chiral, two stereoisomers exist. In a preferred embodiment, the compound represented by general formula (III-P) has R2 in the R configuration. In another preferred embodiment, the compound represented by general formula (III-P) has R2 in the S configuration.

[0083] In a preferred embodiment, R2 represents -OCH2CHF2 according to general formula (III-Q): [ka]

[0084] This structural motif is realized in Examples EX-I-26 and EX-I-27.

[0085] In a preferred embodiment, R3 represents -H.

[0086] In another preferred embodiment, R3 represents -CH3.

[0087] In a preferred embodiment, R4 and R5 each represent -H.

[0088] In other preferred embodiments, R4 and R5 both represent = O.

[0089] In particularly preferred embodiments, the compounds according to the present invention are selected from the group consisting of: [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] and its physiologically acceptable salts.

[0090] In one preferred embodiment, the compound of the present invention takes the form of a free base.

[0091] In another preferred embodiment, the compound in the present invention takes the form of a physiologically acceptable salt.

[0092] For the purposes of this explanation, “salt” should be understood as any form of compound that assumes an ionic form, or is charged and bonded to a counterion (cation or anion), or is dissolved. The term should also be understood as meaning a complex of a compound with other molecules and ions, particularly complexes that associate through ionic interactions. Preferred salts are physiologically acceptable, and are in particular physiologically acceptable salts of anions or acids, or salts formed by physiologically acceptable acids.

[0093] Physiologically acceptable salts of anions or acids are salts of the specific compound in question with an inorganic or organic acid that is physiologically acceptable, particularly when used in humans and / or mammals. Examples of physiologically acceptable salts of a particular acid include, but are not limited to, salts of hydrochloric acid, sulfuric acid, and acetic acid.

[0094] The present invention also includes enantiomers or diastereomers as racemic compounds, or in any enantiomer-excess and diastereomer-excess forms, respectively.

[0095] Furthermore, the present invention also includes isotopic isomers in which at least one atom of the compound in the present invention is replaced with an isotope of a corresponding atom different from the isomer that is dominant in nature, as well as any mixture of such isotopic isomers of the compound. Preferred isotopes are 2 H (Deuterium), 3 H (tritium), 13 C and 14 It is C.

[0096] Further aspects of the present invention relate to the compounds of the present invention as pharmaceuticals.

[0097] Further aspects of the present invention relate to compounds of the present invention for use in the treatment of pain. Further aspects of the present invention relate to a method for treating pain, comprising administering a pain-relieving amount of a compound of the present invention to a subject in need, preferably a human. The pain is preferably acute or chronic. The pain is preferably analgesic or neuropathic.

[0098] Further aspects of the present invention relate to compounds of the present invention for use in the treatment of neurodegenerative diseases, neuroinflammatory diseases, neuropsychiatric disorders, and substance abuse / dependence. Further aspects of the present invention relate to a method for treating any one of the aforementioned disorders, diseases, or conditions, comprising administering a therapeutically effective amount of a compound of the present invention to a subject in need, preferably a human.

[0099] Another aspect of the present invention relates to a pharmaceutical composition comprising a physiologically acceptable carrier and at least one compound of the present invention.

[0100] Preferably, the composition of the present invention is in solid, liquid, or paste form, and / or contains the compound of the present invention in an amount of 0.001 to 99% by weight, preferably 1.0 to 70% by weight, based on the total weight of the composition.

[0101] The pharmaceutical composition of the present invention may optionally contain suitable additives and / or auxiliaries, and / or further active ingredients.

[0102] Suitable physiologically acceptable carriers, additives, and / or auxiliaries include fillers, solvents, diluents, colorants, and / or binders. These substances are known to those skilled in the art (see HP Fiedler, Lexikon der Hilfsstoffe fur Pharmazie, Kosmetik and angrenzende Gebiete, Editio Cantor Aulendoff).

[0103] The pharmaceutical composition of the present invention contains the compound of the present invention in an amount of preferably 0.001 to 99% by weight, more preferably 0.1 to 90% by weight, even more preferably 0.5 to 80% by weight, most preferably 1.0 to 70% by weight, and particularly 2.5 to 60% by weight, based on the total weight of the pharmaceutical composition.

[0104] The pharmaceutical composition according to the present invention is preferably for systemic, topical, or local administration, and preferably for oral administration.

[0105] Another aspect of the present invention relates to a pharmaceutical dosage form containing the pharmaceutical composition of the present invention.

[0106] In a preferred embodiment, the pharmaceutical dosage form of the present invention is formulated for administration twice daily, once daily, or less frequently than once daily. Administration is preferably systemic, particularly orally.

[0107] The pharmaceutical dosage forms in this invention can be administered, for example, as liquid dosage forms in the form of injection solutions, drops, or juices, or as semi-solid dosage forms in the form of granules, tablets, pellets, patches, capsules, plasters / spray plasters, or aerosols. The selection and amount of adjuvants, etc., depend on whether the administration method is orally, perorally, parenterally, intravenously, intraperitoneally, intradermally, intramuscularly, intranasally, buccally, rectally, or locally (for example, to the skin, mucous membranes, or into the eyes).

[0108] Pharmaceutical dosage forms in the form of tablets, dragees, capsules, granules, drops, juices, and syrups are suitable for oral administration, while solutions, suspensions, readily reconstituted dry preparations, and sprays are suitable for parenteral, topical, and inhalation administration. Compounds of the present invention, optionally with the addition of agents that promote skin penetration in depot, dissolved, or plaster forms, are suitable transdermal administration preparations.

[0109] The amount of the compound used in this invention administered to a patient varies depending on the patient's weight, type of administration, indication, and severity of the disease. Typically, at least one compound from this invention is administered at a dose of 0.00005 mg / kg to 50 mg / kg, preferably 0.001 mg / kg to 10 mg / kg.

[0110] Another aspect of the present invention relates to the process for preparing the compounds of the present invention. Preferred processes for synthesizing the compounds of the present invention are, in principle, known to those skilled in the art.

[0111] The preferred synthesis route is described below.

[0112] The compounds in this invention can be obtained through different synthetic routes. Different intermediates are prepared depending on the synthetic route, followed by further reactions.

[0113] According to the first process, the compound of formula (I) may be prepared from the compounds of formula (II) or (VIII) or (XI) and (III), as illustrated by Formula 1. [ka]

[0114] Method 1

[0115] X represents either CR4R5, i.e., CH2 or C=O.

[0116] Hal 1 This is a halogen, preferably Cl, Br, or I.

[0117] The compound of formula (I) may be prepared according to process step (a), a Ullmann-type copper-catalyzed arylation cross-coupling reaction. Typical conditions involve reacting an amine of formula (II), (VIII), or (XI) with a halide of formula (III) in the presence of a suitable copper(II) catalyst, a suitable inorganic base, and a suitable ligand in a suitable solvent and at a suitable temperature. Preferred conditions involve reacting the compounds of formula (II), (VIII), or (XI) and (III) in a sealed tube at 90°C to 180°C in a suitable solvent such as dioxane or DMA in the presence of CuI or Cu2O, DMEDA, and K3PO4 or K2CO3. Alternatively, the compound of formula (I) may be prepared according to process step (b), a Buchwald-Hartwig cross-coupling reaction. Typical conditions involve reacting an amine of formula (II), (VIII), or (XI) with a halide of formula (III) in the presence of a suitable inorganic base and a suitable palladium catalyst in a suitable solvent at a high temperature. Preferred conditions include reacting the compounds of formula (II) or (VIII) or (XI) and (III) in combination with Pd2(dba)3 in the presence of Xantphos, either under reflux or in a sealed container, in the presence of a suitable base such as Cs2CO3, in a suitable solvent such as dioxane at 90°C to 130°C.

[0118] According to the second process, the compound of formula (II) may be prepared from the compounds of formulas (IV), (V), (VI), (VII), and (VIII), as illustrated by Formula 2. [ka]

[0119] Method 2

[0120] Hal 2 is a halogen, preferably Br.

[0121] OR1 and OR2 together are a carbonyl protecting group, preferably 1,3-dioxolane.

[0122] The compound of formula (VI) may also be prepared from the compounds of formulas (IV) and (V) by the organometallic substitution reaction in process step (c). Typical conditions include reacting the halide of formula (IV) with a suitable organometallic base such as a Grignard reagent (preferably iPrMgBr) in a suitable aprotic solvent (preferably THF) at a temperature of 15°C to 50°C, followed by adding the compound of formula (V) in an aprotic solvent such as THF.

[0123] The compound of formula (VII) may also be prepared from the compound of formula (VI) according to the ketal deprotection reaction of process step (d). Typical conditions include reacting the ketal of formula (VI) with a suitable acid (e.g., an aqueous sulfuric acid solution) at a suitable temperature, such as room temperature.

[0124] The compound of formula (VIII) may be prepared from the compound of formula (VII) according to the Bucherer-Bergus reaction of process step (e). Typical conditions involve a reaction between the ketone of formula (VII), potassium cyanide, and ammonium carbonate in aqueous methanol at a suitable temperature of room temperature to about 70°C, preferably at 70°C.

[0125] The compound of formula (II) may be prepared from the compound of formula (VIII) according to the reduction reaction of process step (f). Typical conditions involve the reaction of the compound of formula (VIII) with a suitable reducing agent, preferably BH3·DMS, in a suitable aprotic solvent (preferably THF) at a temperature of 0°C to 65°C.

[0126] According to the third process, the compound of formula (XI) may be prepared from the compounds of formulas (II), (IX), and (X), as illustrated by formula 2. [ka]

[0127] Method 3

[0128] R1 and R2 are protecting groups, preferably R1=R2, and preferably p-methoxybenzyl.

[0129] The compound of formula (IX) may also be prepared from the compound of formula (II) by the alkylation reaction in process step (g). Typical conditions involve the reaction of urea of ​​formula (II) with a suitable alkylating reagent, preferably PMBCl and a suitable base, preferably t-BuOK, in a suitable aprotic solvent, preferably DMSO, at a temperature of 15°C to 50°C.

[0130] The compound of formula (X) may also be prepared from the compound of formula (IX) according to the demethylation reaction of process step (h). Typical conditions include reacting the amine of formula (IX) with a suitable demethylation reagent such as N-iodosuccinimide in a suitable solvent such as MeCN at a suitable temperature such as room temperature.

[0131] The compound of formula (XI) may also be prepared from the compound of formula (X) according to the deprotection reaction of process step (i). Typical conditions include reacting the urea of ​​formula (X) with a suitable acid (e.g., TFA) at a suitable temperature such as 90°C.

[0132] The compounds of formulas (III), (IV), and (V) can be commercially available or prepared by methods known in the literature or similar to those described in the experimental section below.

[0133] The compounds of formulas (I) and (III) can be converted to alternative compounds of formulas (I) and (III) by standard chemical transformations known to those skilled in the art. Examples of these transformations include, but are not limited to, alkylation of heteroatoms such as N or O, halogen interconversion, fluorination, and nucleophilic reactions of ketones.

[0134] Those skilled in the art will understand that it may be necessary to utilize a suitable protecting group strategy to prepare the compounds of formulas (I) and (III).

[0135] Furthermore, it will be understood that in order to obtain the desired compound of the present invention, it may be necessary or desirable to perform the transformations in a different order than that described in the formula, or to modify one or more of the transformations.

[0136] Examples

[0137] "RT" means room temperature (23±7℃), "M" indicates concentration in mol / l units, and "aq." means

[0138] It means aqueous solution, "sat." means saturated, "sol." means solution, and "conc." means concentrated.

[0139] Further abbreviations: Aq. means aqueous, BH3·DMS means boranedimethyl sulfide complex, br means broad, °C means degrees Celsius, CDCl3 means deuterochloroform, Cs2CO3 means cesium carbonate, d means chemical shift, d means double line, dd means twice the double line, DCM means dichloromethane, DABCO means 1,4-diazabicyclo[2.2.2]octane, DAST means diethylaminosulfur trifluoride, DMEDA means 1,2-dimethicone DMF means ethylenediamine, DMF means N,N-dimethylformamide, DMF-DMA means dimethylformamide dimethylacetal, DMSO means dimethyl sulfoxide, DMSO-d6 means hexadeuterodimethyl sulfoxide, Et means ethyl, EtOH means ethanol, SiO means ethyl acetate, Eq means equivalent, g means grams, H2SO4 means sulfuric acid, HCl means hydrochloric acid, HCO2H means formic acid, Hex means hexane. 1H NMR stands for Proton Nuclear Magnetic Resonance, H2O stands for Water, HPLC stands for High Pressure Liquid Chromatography, h stands for Time, IPA stands for 2-Propanol, iPrMgBr stands for Magnesium Isopropyl Bromide, KCN stands for Potassium Cyanide, K2CO3 stands for Potassium Carbonate, K3PO4 stands for Tribasic Potassium Phosphate, L stands for Liter, LCMS stands for Liquid Chromatography Mass Spectrometry, m stands for Multiline, M stands for Molar Concentration, Me stands for Methyl, MeCN stands for Acetonitrile, MeI stands for Iodomethane, MeOH stands for Methanol, MeOH-d4 stands for Deutero-Methanol, Me3SiOK stands for Potassium Trimethylsilanolate, mg stands for Milligram, MgSO4 stands for Magnesium Sulfate, MHz stands for Megahertz, min stands for Min, mL stands for Milliliter, mmol stands for Millimole, MS m / z means mass spectral peak, MTBE means tert-butylmethyl ether, N2 means nitrogen, Na2CO3 means sodium carbonate, NaH means sodium hydride, NaHCO3 means sodium bicarbonate, NaOH means sodium hydroxide, Na2SO4 means sodium sulfate, NH4Cl means ammonium chloride, NH4OH is ammonium hydroxide, (NH4)2CO3 means ammonium carbonate, NIS means N-iodosuccinimide, PE means petroleum ether, q means quadruple, rt is Room temperature (23°C) is indicated, PMBCl means 4-methoxybenzyl chloride, RT means retention time, s means singlet, sat. means saturated, sorn. means solution, t means triplet, TEA means triethylamine, TFA means trifluoroacetic acid, THF means tetrahydrofuran, TLC means thin-layer chromatography, μL means microliter, μmol means micromol, xanthophos means 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene, and XeF2 means xenon difluoride.

[0140] The yields of the prepared compounds were not optimized. All temperatures were not corrected.

[0141] All starting materials not explicitly stated are commercially available (for example, details of suppliers such as Acros, Aldrich, Bachem, Butt Park, Enamine, Fluka, Lancaster, Maybridge, Merck, Sigma, TCI, Oakwood, etc. can be found, for example, in the Symyx® Available Chemicals database at MDL, San Ramon, US or the SciFinder® database at ACS, Washington DC, US), or their synthesis has already been described in detail in the specialized literature (experimental guidelines can be found, for example, in the Reaxys® database at Elsevier, Amsterdam, NL or the SciFinder® database at ACS, Washington DC, US), or they can be prepared using conventional methods known to those skilled in the art.

[0142] Specify the mixing ratio of the chromatography solvent or eluent in v / v.

[0143] All intermediate products and exemplary compounds were analytically characterized by mass spectrometry. In addition, ¹H-NMR and ¹³C spectroscopy were performed on all exemplary compounds and selected intermediate products.

[0144] Regarding stereochemistry and its nomenclature, "cis" refers to the relative stereochemistry of the compounds described herein, where both nitrogen atoms are depicted on the same plane of the cyclohexane ring, as illustrated in the following exemplary structures. Two possible diagrams are possible. [ka]

[0145] Synthesis of intermediates

[0146] INT_001: Synthesis of 8-(3,5-difluorophenyl)-N,N-dimethyl-1,4-dioxaspiro[4.5]decane-8-amine [ka]

[0147] A solution of 1-bromo-3,5-difluorobenzene (321 g, 1.66 mol) in THF (6 L) was slowly added dropwise to a solution of iPrMgBr (832 mL, 2 M) at 15°C, and the mixture was slowly warmed to 50°C. The solution was cooled to 15°C and stirred for 2 hours, after which it was added dropwise to a solution of 8-(dimethylamino)-1,4-dioxaspiro[4.5]decane-8-carbonitrile (100 g, 476 mmol) in THF (600 mL) over 1 hour at 0°C. The resulting mixture was stirred at 15°C for 12 hours. The reaction mixture was slowly added to saturated NH4Cl (1.70 L) and extracted with ELISA (1 L × 2). The combined organic matter was washed with brine (500 mL), dried (Na2SO4), and evaporated to dryness under vacuum to obtain the title compound as oil (157 g, crude). 1 H NMR(400 MHz,CDCl3) δ: 6.84(br d,2H),6.77-6.62(m,1H),3.99-3.90(m,4H),2.26-2.16(m,2H),2.11-2.01(m, 8H), 1.90-1.81 (m, 2H), 1.54-1.46 (m, 2H).

[0148] INT_002: Synthesis of 4-(3,5-difluorophenyl)-4-(dimethylamino)cyclohexane-1-one [ka]

[0149] Two reactions of equal scale were carried out in parallel. 8-(3,5-Difluorophenyl)-N,N-dimethyl-1,4-dioxaspiro[4.5]decan-8-amine (INT_001, 275 g, 925 mmol) was added to H2SO4 (1.36 kg, 1.39 mol, 10% w / w aqueous solution) at 15 °C, and the resulting mixture was stirred at 15 °C for 16 h. The reaction mixture was extracted with DCM (1 L). The aqueous phase was adjusted to pH > 11 by adding saturated NaOH, and the solid was collected by filtration. The filter cake was washed with H2O (500 mL), and the solid was dried under reduced pressure to obtain the title compound as a solid (190 g, crude). 1 H NMR (400 MHz, DMSO-d6) δ: 6.83 - 6.59 (m, 3H), 2.65 - 2.44 (m, 4H), 2.21 (dt, 2H), 2.07 (s, 6H), 2.03 - 1.95 (m, 2H).

[0150] INT_003: Synthesis of cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-1,3-diazaspiro[4.5]decan-2,4-dione

Chemical formula

[0151] Four reactions were carried out in parallel. 4-(3,5-Difluorophenyl)-4-(dimethylamino)cyclohexan-1-one (INT_002, 125 g, 494 mmol) was dissolved in H2O (1.25 L) and MeOH (1.25 L) at 15 °C. (NH4)2CO3 (119 g, 1.23 mol) and KCN (32.4 g, 498 mmol) were added thereto, and the solution was stirred at 70 °C for 16 h. The four parallel reactions were combined, and the solid was collected by filtration. The filter cake was washed with H2O (2 L) and MeOH (2 L), and the solid was triturated with MeOH (500 mL) at 70 °C for 16 h. The solid was recovered and dried under reduced pressure to obtain the title compound as a solid (570 g, crude), which was used without further purification. 1H NMR(400 MHz,DMSO-d6) δ: 8.31-8.19(m,1H),7.16-7.10(m,1H),7.02(br d,2H),3.45-3.20(m,1H),2.46-2.37(m, 2H),2.08-1.84(m, 8H),1.55(br d, 2H),1.47-1.30(m, 2H).

[0152] INT_004: Synthesis of cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-1,3-diazaspiro[4.5]decan-2-one [ka]

[0153] Two reactions were carried out in parallel. BH3·DMS (711 mL, 10 M) was added at 0°C to a solution of cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-1,3-diazaspiro[4.5]decane-2,4-dione (INT_003, 230 g, 711 mmol) in THF (1.3 L), and the resulting mixture was stirred at 65°C for 16 hours. The reaction mixture was very slowly added to HCl (6 M, 1 L), followed by MeOH (1 L) and H2O (1 L), and stirred for 10 minutes. The solid was removed by filtration, and the filtration cake was washed with H2O (500 mL) and MeOH (500 mL). The pH of the filtrate was adjusted to above pH 13 by adding saturated NaOH. The solid was collected by filtration, the filtration cake was washed with H2O (2 L), and dried under reduced pressure to obtain the residue. The residue was added to HCl (500 mL, 6 M), stirred for 10 minutes, then collected as a solid and washed with H2O (100 mL). The solid was again added to H2O (150 mL), stirred for 10 minutes, collected by filtration, and washed with H2O (100 mL). This process was repeated, and the collected solid was dried under reduced pressure. The solid was added to H2O (500 mL), stirred for 2 minutes, and then the pH was adjusted with saturated NaOH until it was above pH 12. Stirring was continued for 10 minutes, the solid was collected by filtration, washed with H2O (500 mL), and dried under reduced pressure to obtain the title compound as a solid (200 g, 647 mmol). 1H NMR(400 MHz,DMSO-d6) δ: 7.12(t,1H),7.07-6.99(m,2H),6.60(br s,1H),6.02(s,1H),3.01(s,2H)), 2.24(br s, 2H), 1.94(s, 6H), 1.77(br d, 4H), 1.44-1.27(m, 2H).

[0154] INT_005: Synthesis of 2-bromo-5-cyclopropoxy-3-methylpyridine [ka]

[0155] Twenty-four corresponding reactions were carried out in parallel. A mixture of 6-bromo-5-methylpyridine-3-ol (1.50 g, 7.98 mmol), Cs2CO3 (3.38 g, 10.4 mmol), KI (199 mg, 1.20 mmol), and bromocyclopropane (3.86 g, 31.9 mmol) in DMF (10 mL) was stirred at 140 °C for 2 hours under microwave conditions. The 24 reaction products were combined, the solid was removed by filtration, and the filtration cake was washed with HCl (50 mL). The filtrate was extracted with HCl (80 mL x 4), and the combined organic matter was evaporated to dryness under vacuum. The residue was purified by column chromatography (SiO2, 25-100% HCl / PE) to obtain the title compound as a solid (7.00 g, yield 15.7%). 1 H NMR(400 MHz,DMSO-d6) δ: 8.03(d,1H),7.53(d,1H),3.95(tt,1H),2.31(s,3H),0.87-0.65(m,4H).

[0156] INT_006: Synthesis of 5-bromo-2-methoxy-4-methylpyrimidine [ka]

[0157] To a stirred solution of 5-bromo-2-chloro-4-methylpyrimidine (5 g, 24.1 mmol) in THF (120 mL), NaOMe (25% in MeOH, 6.05 mL, 26.49 mmol) was added at 0°C and the mixture was stirred at room temperature for 16 hours. The reaction mixture was cooled to 0°C, slowly quenched with saturated NH4Cl aqueous solution, and extracted with RINKAN (200 mL). The combined organic matter was washed with water (100 mL) and brine (100 mL), dried in (Na2SO4), and evaporated to dryness under vacuum. The residue was purified by combiflash column chromatography (0-5% RINKAN / Hexane) to obtain the title compound (3.6 g, 73%) as oil. LCMS m / z = 205 [M+H] + .

[0158] INT_007: Synthesis of 2-(6-chloro-5-methylpyridine-3-yl)propan-2-ol [ka]

[0159] iPrMgCl.LiCl (1.30 M, 117 mL) was added dropwise to a mixture of THF (500 mL), molecular sieves (5 Å, 10 g) and 5-bromo-2-chloro-3-methylpyridine (30.0 g, 145 mmol) at 0°C. The mixture was stirred at 0°C for 1 hour. Acetone (12.7 g, 218 mmol) was added dropwise to the reaction mixture over 1 hour at 0°C, and the mixture was stirred at 0°C for 1 hour. The reaction mixture was quenched with NH4Cl (500 mL) and extracted with HCl (3 × 500 ml). The combined organic matter was washed with brine, dried with (Na2SO4), and evaporated to dryness under vacuum. The residue was purified by column chromatography (SiO2, 50:1-0:1 PE / HCl) to obtain the title compound (15 g, 53%) as oil. LCMS m / z = 186 [M+H] + .

[0160] INT_008: Synthesis of 2-bromo-5-(difluoromethoxy)-3-methylpyridine [ka]

[0161] A mixture of 6-bromo-5-methylpyridine-3-ol (31.0 g, 165 mmol), sodium 2-chloro-2,2-difluoroacetate (126 g, 829 mmol), Cs2CO3 (39.2 g, 120 mmol), and K2CO3 (16.6 g, 120 mmol) in DMF (310 mL) was stirred at 100 °C for 16 hours. The reaction mixture was filtered, and the filtrate cake was diluted with ethyl acetate. The filtrate was extracted with ethyl acetate (250 mL x 2) and water (300 mL). The combined organic matter was washed with brine (200 mL x 2), dried under reduced pressure, and evaporated to dryness. The residue was purified by column chromatography (SiO2, 5:1 PE / ethyl acetate) to obtain the title compound as oil (16.8 g, 42.8%). 1 H NMR (400 MHz, CDCl3) δ: 8.11 (s, 1H), 7.40-7.34 (m, 1H), 6.77-6.35 (m, 1H), 2.42 (s, 3H).

[0162] INT_009: Synthesis of 3-bromo-4-methyl-5-nitropyridine-2(1H)-one [ka] To a stirred solution of 4-methyl-5-nitro-1,2-dihydropyridine-2-one (6.0 g, 38.961 mmol) in AcOH (30 ml), bromine (2.4 ml, 46.7 mmol) was added at room temperature, and the mixture was stirred at that temperature for 16 hours. The reaction mixture was poured onto ice, and the product precipitated as a white solid. The product was recovered by filtration, washed with H2O, and the title compound was obtained as a solid (8 g, 88%). LCMS m / z = 233 [M+H] + .

[0163] INT_010: Synthesis of 3-bromo-2-chloro-4-methyl-5-nitropyridine [ka] To a stirred MeCN solution of 3-bromo-4-methyl-5-nitro-1,2-dihydropyridine-2-one (INT_009, 5.0 g, 21.5 mmol), DIPEA (3.7 ml, 21.5 mmol) and POCl3 (20.0 ml, 214 mmol) were added at 0°C. The reaction mixture was stirred at 80°C for 3 hours. The reaction mixture was concentrated under reduced pressure, 30 ml of ice-cold water was added, and the resulting mixture was made basic by adding a saturated aqueous solution of NaHCO3. This mixture was extracted with siRNA (60 mL). The organic layer was dried over anhydrous Na2SO4 and concentrated under vacuum. The residue was purified by column chromatography to obtain the title compound as a solid (4.3 g, 79%). LCMS m / z = 251 [M+H] + .

[0164] INT_011: Synthesis of (E)-2-(3-bromo-2-chloro-5-nitropyridine-4-yl)-N,N-dimethylethene-1-amine [ka] A solution of 3-bromo-2-chloro-4-methyl-5-nitropyridine (INT_010, 4.3 g, 17.1 mmol) in DMF-DMA (25 mL) was stirred at 50°C for 16 hours. The reaction mixture was concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, 20% Â / hexane) to obtain the title compound (5.0 g, 95%) as a solid. LCMS m / z = 306 [M+H] + .

[0165] INT_012: Synthesis of 4-bromo-5-chloro-1H-pyrrolo[2,3-c]pyridine [ka] To a solution of [(E)-2-(3-bromo-2-chloro-5-nitropyridine-4-yl)ethenyl]dimethylamine (INT_011, 5g, 16.3 mmol) in EtOH and H2O, Fe (4.6g, 81.6 mmol) and NH4Cl (8.7g, 163.1 mmol) were added, and the reaction mixture was stirred at 100°C for 2 hours. The reaction mixture was filtered, concentrated under reduced pressure, and diluted with HCl (60 ml). This solution was washed with water (30 ml) and saturated NaCl solution (30 mL). The organic layer was dried over anhydrous Na2SO4 and concentrated under vacuum. The residue was purified by column chromatography (SiO2, 20% HCl / hexane) to obtain the title compound (1.6g, 42%) as a solid. LCMS m / z = 231 [M+H] + .

[0166] INT_013: Synthesis of 5-chloro-4-methyl-1H-pyrrolo[2,3-c]pyridine [ka] To a stirred solution of 4-bromo-5-chloro-1H-pyrrolo[2,3-c]pyridine (INT_012, 1.0 g, 4.32 mmol) with 1,4-dioxane and H2O, trimethylboroxine (0.73 ml, 5.2 mL), K2CO3 (1.791 g, 12.959 mmol, 3 equivalents), and Pd(dppf)Cl2 (0.32 g, 0.43 mmol) were added. The reaction mixture was stirred at 90°C for 16 hours and filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography (SiO2, 35% siRNA / hexane) to obtain the title compound (0.3 g, 42%) as a solid. LCMS m / z = 167 [M+H] + .

[0167] INT_014: Synthesis of 5-chloro-4-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-c]pyridine [ka] To a stirred solution of 5-chloro-4-methyl-1H-pyrrolo[2,3-c]pyridine (INT_013, 0.15 g, 0.9 mmol) in DMF (5 mL), NaH (60% in mineral oil, 0.07 g, 1.8 mmol) was added at 0°C and the mixture was stirred for 20 minutes. SEM-Cl (0.24 ml, 1.35 mmol) was added to the reaction mixture and the mixture was stirred at room temperature for 1 hour. The reaction mixture was quenched with aqueous NH4Cl solution, extracted with siRNA (30 mL), and washed with H2O (10 mL) and saturated NaCl solution (10 mL). The combined organic layers were dried over anhydrous Na2SO4 and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, 10% siRNA / hexane) to obtain the title compound (0.2 g, 74%). LCMS m / z = 297 [M+H] + .

[0168] INT_015: Synthesis of cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-3-(4-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,3-c]pyridine-5-yl)-1,3-diazaspiro[4.5]decan-2-one [ka]

[0169] To a stirred solution of cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-1,3-diazaspiro[4.5]decan-2-one (INT_004, 0.15 g, 0.49 mmol) in 1,4-dioxane (5 mL), 5-chloro-4-methyl-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrrolo[2,3-c]pyridine (INT_014, 0.173 g, 0.58 mmol) and K2CO3 (0.2 g, 1.5 mmol) were added. The mixture was purged with argon and degassed for 5 minutes. Then, trans-N,N'-dimethylcyclohexane-1,2-diamine (0.015 mL, 0.096 mmol) and CuI (0.009 g, 0.048 mmol) were added, and the resulting mixture was stirred at 130 °C for 16 hours. The reaction mixture was cooled to room temperature and filtered through a Celite pad. The filtrate was concentrated under reduced pressure, and the residue was diluted with 10% MeOH in DCM (50 mL), then washed with water (20 mL) and saturated NaCl solution (20 mL). The organic layer was dried over anhydrous Na2SO4 and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, 2-3% MeOH / DCM) to obtain the title compound (0.2 g, 72%). LCMS m / z = 570 [M+H] + .

[0170] INT_016: Synthesis of cis-(3,5-difluorophenyl)-8-(dimethylamino)-1,3-bis(4-methoxybenzyl)-1,3-diazaspiro[4.5]decan-2-one [ka]

[0171] A solution of cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-1,3-diazaspiro[4.5]decan-2-one (INT_004, 10.0 g, 32.2 mmol) in DMSO (100 mL) was added with t-BuOK (10.8 g, 96.7 mmol) and PMB-Cl (13.0 mL, 19.7 mmol) at room temperature, and the mixture was stirred at that temperature for 2 hours. The reaction was quenched by adding H2O (150 mL) and extracted with EtOAc (2 × 150 mL). The combined organic layers were washed with saturated NaCl solution (100 ml), dried over anhydrous Na2SO4, and concentrated in vacuo. The residue was purified by column chromatography (SiO2, 20 - 30% EtOAc / PE) to give the title compound (14.0 g, 79%) as a solid. LCMS m / z = 550.9 [M+H] + .

[0172] INT_017: Synthesis of cis-8-(3,5-difluorophenyl)-1,3-bis(4-methoxybenzyl)-8-(methylamino)-1,3-diazaspiro[4.5]decan-2-one

Chemical Structure

[0173] A solution of cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-1,3-bis(4-methoxybenzyl)-1,3-diazaspiro[4.5]decan-2-one (INT_016, 17.1 g, 31.1 mmol) in MeCN (340 mL) was added with NIS (10.5 g, 46.6 mmol) and stirred at room temperature for 1 hour. The reaction was quenched by adding a dilute aqueous solution of Na2S2O3 (100 mL) and extracted with EtOAc (2 × 150 mL). The combined organic layers were washed with saturated NaCl solution (100 ml), dried over anhydrous Na2SO4, and concentrated in vacuo. The residue was purified by column chromatography (SiO2, 30 - 40% EtOAc / PE) to give the title compound (13.1 g, 79%) as a solid. LCMS m / z = 536.9 [M+H] + .

[0174] INT_018: Synthesis of cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-1,3-diazaspiro[4.5]decan-2-one [Chemical formula]

[0175] Dissolve cis-8-(3,5-difluorophenyl)-1,3-bis(4-methoxybenzyl)-8-(methylamino)-1,3-diazaspiro[4.5]decan-2-one (INT_017, 13.0 g, 24.3 mmol) in TFA (130 mL) and stir at 90 °C for 18 hours. Concentrate this reaction mixture under reduced pressure, wash with EtOAc (100 mL), make it basic with an aqueous NaHCO3 solution (500 mL), and extract with 10% MeOH / DCM (2 × 500 mL). Wash the combined organic layers with a saturated NaCl solution (200 ml), dry over anhydrous Na2SO4, and concentrate in vacuo to obtain the title compound (6.0 g, 84%) as a solid. LCMS m / z = 296.2 [M+H] + .

[0176] INT_019: Synthesis of 5-bromo-2-cyclopropoxy-4-methylpyrimidine [Chemical formula]

[0177] Cyclopropyl alcohol (15.1 g, 260 mmol) in THF (40 mL) was added dropwise to a suspension of NaH (10.4 g, 260 mmol) in THF (800 mL) over 30 minutes at 0°C, and the mixture was stirred at 0°C for 30 minutes. A solution of 5-bromo-2-chloro-4-methylpyrimidine (45.0 g, 217 mmol) in THF (80 mL) was added dropwise over 30 minutes at 0°C, and the resulting mixture was stirred at 25°C for 16 hours. The reaction product was quenched with saturated Na₄Cl aqueous solution (1 L) at 10°C, and then extracted with siRNA (3 × 400 mL). The combined organic matter was dried and evaporated to dryness under reduced pressure. The residue was purified by column chromatography (SiO₂, 16-100% siRNA / PE) to obtain the title compound as oil (19.0 g, 38%). 1 H NMR (400 MHz, DMSO-d6) δ: 8.67(s,1H),4.34-4.21(m,1H),2.49(s,3H),0.81-0.75(m,2H),0.74-0.68(m,2H).

[0178] INT_020: Synthesis of 4-bromo-1,3-dimethylpyridine-2(1H)-one [ka]

[0179] MeI (16.6 mL, 266 mmol) was added at room temperature under argon to a solution of 4-bromo-3-methylpyridine-2(1H)-one (1) (5.0 g, 26.6 mmol) and t-BuOK (5.96 g, 53.2 mmol) in 1,4-dioxane (250 mL), and the mixture was heated at 80°C for 18 hours. The reaction mixture was cooled to room temperature, filtered through a Celite pad, and washed with DCM (2 × 20 mL). The filtrate was evaporated to dryness under vacuum, and the residue was purified by flash chromatography (70-80% siRNA / PE) to obtain the title compound as a solid (5.3 g, 98%). LCMS m / z = 202 [M+H] + .

[0180] INT_021: Synthesis of 2-(5-chloro-6-methylpyrazine-2-yl)propan-2-ol [ka]

[0181] To a stirred THF (100 mL) solution of methyl 5-chloro-6-methylpyrazine-2-carboxylate (3.5 g, 18.75 mmol), MeMgBr (3 M Et2O; 19 ml, 56.25 mmol) was added dropwise at 0°C, and the mixture was stirred at 0°C for 2 hours. The reaction product was quenched with a saturated aqueous solution of NH4Cl (50 mL) and extracted with siRNA (2 × 80 mL). The combined organic matter was dried (Na2SO4) and evaporated to dryness under vacuum. The residue was purified by column chromatography (SiO2, 0-20% siRNA / Hex) to obtain the title compound (2.2 g, 63%) as oil. LCMS m / z = 187 [M+H] + .

[0182] INT_022: Synthesis of 5-bromo-2-iodo-4-methylpyrimidine [ka]

[0183] To a 10 mL solution of 5-bromo-2-chloro-4-methylpyrimidine (1.0 g, 4.83 mmol) in DCM, 0.92 mL of HI aqueous solution (20.77 mmol) was added at room temperature, and the mixture was stirred at room temperature for 16 hours. The reaction mixture was quenched with ice water (50 mL) and extracted with ethyl acetate (2 × 100 mL). The combined organic matter was washed with water (50 mL) and brine solution (50 mL), dried, and evaporated to dryness under vacuum. The residue was purified by column chromatography (SiO2, 0-10% ethyl acetate / PE) to obtain the title compound (900 g, 63%) as a solid. LCMS m / z = 299 [M+H] + .

[0184] INT_023: Synthesis of 3-(5-bromo-4-methylpyrimidin-2-yl)oxetan-3-ol

Chem.

[0185] To a solution of 5-bromo-2-iodo-4-methylpyrimidine (INT_022, 0.900 g, 3.03 mmol) in toluene (25 mL) was added n-butyllithium (1.98 mL, 3.18 mmol, 1.6 M in n-hexane) at -78 °C, and the reaction mixture was stirred at -78 °C for 30 minutes. Oxetan-3-one (0.213 mL, 3.64 mmol) was added to the reaction mixture at -78 °C, and the reaction mixture was stirred at -78 °C for 30 minutes. The reaction was quenched with saturated sodium sulfate solution and extracted with EtOAc (2 × 50 mL). The combined organics were washed with H2O (50 mL), brine (50 mL), dried (Na2SO4), and evaporated to dryness in vacuo. The residue was purified by column chromatography (SiO2, 0 - 10% EtOAc / PE) to afford the title compound (300 mg, 40%) as an oil. LCMS m / z = 245 [M+H] + .

[0186] INT_024: Synthesis of 5-bromo-2-(3-fluorooxetan-3-yl)-4-methylpyrimidine

Chem.

[0187] 3-(5-bromo-4-methylpyrimidine-2-yl)oxetan-3-ol (INT_O23, 400 mg, 1.65 mmol) was dissolved in DCM (10 mL), to which DAST (0.4 mL, 3.29 mmol) was added at 0°C. The mixture was stirred at room temperature for 1 hour. The reaction mixture was quenched with ice water (20 mL) and extracted with DCM (2 × 500 mL). The combined organic matter was washed with saturated sodium bicarbonate solution (50 mL), dried (Na2SO4), and evaporated to dryness under reduced pressure. The residue was purified by column chromatography (SiO2, 0-10% Â / PE) to obtain the title compound (240 mg, 59%) as oil. 1 H NMR(400 MHz, CDCl3δ:8.79(s, 1H),5.20-5.00(m, 4H),2.70(s, 3H).

[0188] INT_025: Synthesis of 5-bromo-4-methylpyrimidine-2-carbonitride [ka]

[0189] DABCO (12.24 g, 109 mmol) was followed by NaCN (4.28 g, 87.4 mmol) in a solution of 5-bromo-2-chloro-4-methylpyrimidine (15 g, 72.8 mmol) in DMSO (150 mL) and H2O (150 mL). The mixture was stirred at room temperature for 16 hours. The reaction mixture was extracted with siRNA (2 × 250 mL), washed with H2O (2 × 250 mL) and brine (2 × 250 mL), dried, and evaporated to dryness under vacuum. The residue was purified by combiflash chromatography (5-10% siRNA / Hex) to obtain the title compound (11.5 g, 80%) as a solid. LCMS m / z = 198 [M+H] + .

[0190] INT_026: Synthesis of methyl 5-bromo-4-methylpyrimidine-2-carboxylate [ka]

[0191] A reaction mixture of 5-bromo-4-methylpyrimidine-2-carbonitride (INT_025, 1 g, 5.05 mmol) was stirred in MeOH (12 mL), to which concentrated HCl (12 mL) was added. The reaction mixture was stirred at 80°C for 8 hours. The reaction mixture was poured into a cold saturated NaHCO3 solution and extracted with RINKAN (2 × 50 mL). The combined organic layer was washed with H2O (2 × 25 mL) and brine (2 × 25 mL), dried, and concentrated under reduced pressure in (Na2SO4). The residue was purified by combiflash chromatography (10-50% RINKAN / Hex) to obtain the title compound (434 mg, 37%) as a solid. LCMS m / z = 233 [M+H] + .

[0192] INT_027: Synthesis of 2-(5-bromo-4-methylpyrimidine-2-yl)propan-2-ol [ka]

[0193] The title compound was prepared as a solid (0.25 g, 62%) from methyl 5-bromo-4-methylpyrimidine-2-carboxylate (INT_026) using a method similar to that described for INT_20. LCMS m / z = 231 [M+H] + .

[0194] INT_028: Synthesis of 6-chloro-3-(1,1-difluoroethoxy)-2-methoxypyridine [ka]

[0195] To a solution of 6-chloro-2-methoxynicotinaldehyde (40.0 g, 233 mmol) in THF (800 mL), MeMgBr (3.0 M, 117 mL) was added at 0°C, and the mixture was stirred at that temperature for 30 minutes. The reaction was quenched by adding a saturated aqueous solution of NH4Cl, and the mixture was extracted with siRNA (2 × 800 ml). The combined organic layers were dried over anhydrous Na2SO4 and evaporated to dryness under reduced pressure to obtain the corresponding crude alcohol (48.0 g).

[0196] The residue (25.0 g, 133 mmol) was dissolved in MeCN (375 mL) and cooled to 0°C under a nitrogen atmosphere. Subsequently, ice-cold solutions of NaIO4 (57.0 g, 266 mmol) and RuCl3 (691 mg, 3.33 mmol) in H2O (300 mL) were added dropwise to the organic solution, and the mixture was stirred at room temperature for 1 hour. H2O (300 mL) was added, and the mixture was extracted with RINKAN (2 × 300 mL). The combined organic layers were dried over anhydrous Na2SO4 and concentrated under vacuum to obtain the corresponding ketone as a solid (18.5 g, crude).

[0197] To a 60 mL solution of the crude residue (2.0 g, 10.8 mmol) in DCM, HF-pyridine (12.8 g, 129 mmol) was added at room temperature, and the mixture was stirred for 1 hour. XeF2 (5.47 g, 32.3 mmol) was added, and the mixture was stirred at room temperature for 12 hours. The reaction product was quenched by adding water (15 mL), and the pH was adjusted to 8-9 by adding saturated Na2CO3 aqueous solution. The mixture was extracted with siRNA (3 × 15 mL), and the mixed organic layer was dried on anhydrous Na2SO4 and concentrated under reduced pressure. The residue was purified by column chromatography (SiO2, 1-100% siRNA / PE) to obtain the title compound (1.17 g, yield 48%) as oil. 1 H NMR(400 MHz, CDCl3δ:1.92 - 2.00(m, 3 H)3.98 - 4.01(m, 3 H)6.88(d, 1 H)7.46(d, 1 H).

[0198] INT_029: Synthesis of 6-chloro-3-(1,1-difluoroethoxy)-1-methylpyridine-2(1H)-one [ka]

[0199] To a solution of 6-chloro-3-(1,1-difluoroethoxy)-2-methoxypyridine (INT_O28, 4.5 g, 20.1 mmol) in MeCN (200 mL), NaI (15.1 g, 101 mmol) and TMSCl (10.9 g, 101 mmol) were added at 0°C under a nitrogen atmosphere. The reaction mixture was heated to 70°C and stirred at that temperature for 2 hours, after which it was filtered through a diatomaceous pad. The pad was washed with 1:10 MeOH / DCM (500 mL), and the combined organic layer was concentrated under vacuum. The residue was purified by column chromatography (SiO2, 1-100% ELISA / PE) to obtain an O-demethylation intermediate as a solid (4.12 g).

[0200] To a solution of the above intermediate (2.15 g, 10.3 mmol) in acetone (10 mL), CH3I (1.53 g, 10.8 mmol) and K2CO3 (2.84 g, 20.5 mmol) were added, and the mixture was purged with N2 to degas it. The mixture was stirred at room temperature for 16 hours, and then quenched with H2O (10.0 mL). The mixture was extracted with DCM (10 × 10 mL), and the combined organic layers were dried over Na2SO4 and concentrated under vacuum. The residue was purified by column chromatography (SiO2, 1-100% siRNA / PE) to obtain the title compound (1.49 g, yield 65%) as oil. 1 H NMR(400 MHz, CDCl3δ: 1.98 (t, 3 H)3.71 - 3.73(m, 3 H)6.25(d, 1 H)7.23(d, 1 H).

[0201] INT_030: Synthesis of 1-(6-chloro-5-methylpyridine-3-yl)ethane-1-ol [ka]

[0202] To a solution of 6-chloro-5-methylnicotinaldehyde (1.0 g, 6.426 mmol, 1.0 equivalent) in tetrahydrofuran (20 mL), MeMgBr (3.0 M Et2O, 6.4 mL, 1.28 mmol) was added at 0°C, and the reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was quenched with saturated NH4Cl aqueous solution (10 mL), diluted with ice water (100 mL), and extracted with siRNA (2 × 250 mL). The combined organic layer was washed with H2O (50 mL) and saturated NaCl solution (50 mL), dried over anhydrous Na2SO4, and the solvent was removed under vacuum. The crude product was purified by column chromatography (SiO2, 10% siRNA / PE) to obtain the title compound (0.74 g, 67%) as a solid. LCMS m / z = 172 [M+H] + .

[0203] INT_031: Synthesis of 2-(6-chloro-5-methylpyridine-3-yl)-1,1,1-trifluoropropan-2-ol [ka]

[0204] To a 10 mL solution of 1-(6-chloro-5-methylpyridine-3-yl)ethane-1-ol (INT_030, 0.740 g, 4.327 mmol) in DCM (1.86 g, 8.654 mmol) at room temperature, the reaction mixture was stirred at that temperature for 2 hours. The reaction mixture was filtered through a Celite pad, and the filtered cake was washed with ELISA (200 mL). The filtrate was concentrated under vacuum. The crude product was purified by column chromatography (SiO2, 16% ELISA / PE) to obtain the corresponding ketone (0.60 g) in liquid form.

[0205] To a solution of the above ketone (0.50 g, 2.958 mmol) in THF (25 mL), trimethyl(trifluoromethyl)silane (0.87 mL, 5.917 mmol) and tetra-n-butylammonium fluoride (1.0 M in THF, 0.6 mL, 0.59 mmol) were added at 0°C, and the reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was diluted with ice water (100 mL) and extracted with ethyl acetate (2 × 250 mL). The combined organic layer was washed with H₂O (50 mL) and saturated NaCl solution (50 mL), dried over anhydrous Na₂SO₄, and the solvent was removed under vacuum. The residue was purified by column chromatography (SiO₂, 8% ethyl acetate / PE) to obtain the title compound (0.54 g, 76%) as a solid. LCMS m / z = 240 [M + H] + .

[0206] INT_032: Synthesis of 2-chloro-5-(2,2-difluoroethoxy)-3-methylpyridine [ka] To a stirred solution of 6-chloro-5-methylpyridine-3-ol (0.3 g, 2.1 mmol) in DMF (5.0 mL), Cs2CO3 (1.0 g, 3.1 mmol) and 2,2-difluoroethyltrifluoromethanesulfonate (0.49 mg, 2.3 mmol) were added at room temperature. The reaction mixture was stirred at the same temperature for 2 hours. The reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (2 × 50 mL). The combined organic layers were washed with saturated NaCl solution (50 mL), dried over anhydrous Na2SO4, and concentrated under vacuum to obtain the title compound (0.4 g, 92%) as a solid, which was used without further purification.

[0207] INT_033: Synthesis of 3-chloro-6-(2,2-difluoroethoxy)-4,5-dimethylpyridazine [ka]

[0208] 3,6-dichloro-4,5-dimethylpyridazine (0.20 g, 1.1 mmol) was stirred in THF (10 mL) and NaH (0.09 g, 2.2 mmol) was added at 0°C. The resulting reaction mixture was stirred for 10 minutes, and then 2,2-difluoroethane-1-ol (0.09 g, 1.1 mmol) was added. The reaction mixture was stirred at 90°C for 16 hours. The reaction product was quenched with ice water (10 mL), extracted with siRNA (3 × 20 mL), and washed with water (20 mL). The combined organic layers were dried over anhydrous Na₂SO₄, filtered, and concentrated under vacuum. The crude product was purified by column chromatography (SiO₂, 0-7% siRNA / PE) to obtain the title compound (0.19 mg, 75%) as a solid. LCMS m / z = 223.1 [M+H] + .

[0209] INT_034: Synthesis of 5-(difluoromethoxy)-2-iodo-3-methylpyridine [ka]

[0210] CuI (0.29 g, 1.5 mmol) and DMEDA (133 mg, 1.5 mmol) were added to a 1,4-dioxane (12 mL) stirring solution containing 2-bromo-5-(difluoromethoxy)-3-methylpyridine (INT_008, 0.36 g, 1.5 mmol, 1.0 equivalent) and NaI (1.13 g, 7.6 mmol, 5.0 equivalents) in a high-pressure tube at room temperature under argon. The tube was sealed, the reaction mixture was stirred at 120 °C for 18 hours, then cooled to room temperature, filtered through a Celite pad, and washed with 10% MeOH in siRNA (2 × 25 mL). The filtrate was washed with water (30 mL), dried on anhydrous Na₂SO₄, filtered, and concentrated under vacuum. The crude product was purified by column chromatography (SiO₂, 0-30% siRNA / PE) to obtain the title compound (0.17 g, 41%) in liquid form. LCMS m / z = 285.9 [M+H] + .

[0211] Synthesis of Exemplary Compounds

[0212] EX-I-01: Synthesis of cis-3-(5-cyclopropoxy-3-methylpyridine-2-yl)-8-(3,5-difluorophenyl)-8-(dimethylamino)-1,3-diazaspiro[4.5]decan-2-one [ka]

[0213] A mixture of 2-bromo-5-cyclopropoxy-3-methylpyridine (INT_005, 7.5 g, 32.9 mmol), cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-1,3-diazaspiro[4.5]decan-2-one (INT_004, 12.3 g, 29.9 mmol), and K3PO4 (19 g, 89.7 mmol) in dioxane (600 L) was stirred at 25°C for 30 minutes. CuI (5.69 g, 29.9 mmol) and DMEDA (5.27 g, 59.8 mmol) were added, and the mixture was stirred at 120°C for 16 hours. Solid matter was removed by filtration, the filtrate was washed with MTBE (80 mL), the filtrate was extracted with siRNA (300 mL x 2), and washed with H2O (1000 mL). The combined organic matter was washed with brine (100 mL x 2), dried (Na2SO4), and evaporated to dryness under vacuum. The residue was stirred with MTBE at 15°C for 30 minutes, the solid was collected by filtration, and washed with MTBE (50 mL). The solid was dried under reduced pressure and further purified by grinding. The solid was stirred with MeCN (260 mL) at 90°C for 30 minutes, cooled to 50°C, diluted with MeCN (130 mL), and stirred at 90°C for 30 minutes. The mixture was filtered, the filtrate was cooled to 50°C, stirred at 90°C for 10 minutes, and then cooled to 15°C. The solid was recovered by filtration and dried under reduced pressure to obtain the title compound as oil (23 g, 83.4%). LCMS m / z = 457 [M+H] + ; 1H NMR(400 MHz, DMSO-d6) δ: 7.99 (d, 1H), 7.39 (d, 1H), 7.28 (br s, 1H), 7.13 (br t, 1H), 7.05 (br d, 2H), 3.91 (tt, 1H), 3.58 (d, 2H), 2.39-2.26 (m, 2H), 2.23 (s, 3H), 1.96 (s, 6H), 1.94-1.69 (m, 4H), 1.49(br s, 2H),0.83-0.64(m, 4H).

[0214] EX-I-02: Synthesis of cis-8-(3,5-difluorophenyl-8-(dimethylamino)-3-(2-methoxy-4-methylpyrimidine-5-yl)-1,3-diazaspiro[4.5]decan-2-one [ka]

[0215] 5-bromo-2-methoxy-4-methylpyrimidine (INT_006, 3.54 g, 17.45 mmol) and K2CO3 (7.2 g, 52.35 mmol) were added to a sealed tube containing a solution of cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-1,3-diazaspiro[4.5]decan-2-one (INT_004, 5.45 mmol) in dioxane (70 mL). The mixture was degassed with argon for 15 minutes, then DMEDA (0.52 mL, 3.49 mmol) and CuI (0.332 g, 1.745 mmol) were added, and the mixture was stirred at 130°C for 16 hours. A second batch was carried out using 7 g of INT_004 with the same procedure as above. After the reaction was complete, the mixtures were combined and filtered through a Celite pad. The filtrate was concentrated under vacuum, diluted with 10% MeOH-DCM (400 mL), and washed with water (2 × 150 mL) and brine (150 mL). The combined organic matter was dried (Na₂SO₄) and evaporated to dryness under vacuum. The residue was purified by combiflash chromatography (0-10% MeOH / DCM) to obtain the title compound (9.2 g, 53%) as a solid. LCMS m / z = 432 [M+H]+ ; 1 H NMR(400 MHz, DMSO-d6) δ: 8.38 (s, 1H), 7.34 (brs, 1H), 7.14-7.10 (m, 1H),7.05 (d, 2H), 3.87(s, 3H),3.47(s, 2H), 2.45-2.30(m, 5H), 1.96(s, 6H), 1.93-1.79(m, 4H), 1.53-1.51(m, 2H).

[0216] EX-I-03: Synthesis of cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-3-(5-(2-hydroxypropan-2-yl)-3-methylpyridine-2-yl)-1,3-diazaspiro[4.5]decan-2-one [ka]

[0217] Pd2(dba)3 (11.85 g, 13 mmol) and xanthophos (15.0 g, 25.8 mmol) were added at room temperature under argon to a mixture of cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-1,3-diazaspiro[4.5]decan-2-one (INT_004, 40 g, 129 mmol), 2-(6-chloro-5-methylpyridine-3-yl)propan-2-ol (INT_007, 29 g, 155 mmol), and Cs2CO3 (127 g, 388 mmol) in 1,4-dioxane (1 L). The mixture was stirred at 120 °C for 16 hours, then cooled to room temperature and filtered through a Celite pad. The cake was washed with ELISA (2 × 500 mL), and the combined organic layers were concentrated under reduced pressure. The crude product was purified by combiflash chromatography (5% MeOH / DCM), and the product was recrystallized using MeCN / H2O4:1 (840 mL, 80°C → room temperature) to obtain the title compound as a solid (25 g, 43%). LCMS m / z = 459 [M+H] + ; 1H NMR(400 MHz, DMSO-d6) δ: 8.27(d, 1H), 7.66 (d, 1H), 7.31(br s, 1H), 7.16-7.01(m, 3H), 5.13(s, 1H), 3.66-3.55(m, 2H), 2.37-2.17(m, 5H), 2.01-1.77(m, 10H), 1.57-1.38(m, 8H).

[0218] EX-I-04: Synthesis of cis-3-(5-(difluoromethoxy)-3-methylpyridine-2-yl)-8-(3,5-difluorophenyl)-8-(dimethylamino)-1,3-diazaspiro[4.5]decan-2-one [ka]

[0219] K2CO3 (38.2 g, 276 mmol), CuI (11.7 g, 61.4 mmol), and DMEDA (10.8 g, 123 mmol) were added at 20°C to a mixture of cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-1,3-diazaspiro[4.5]decan-2-one (INT_004, 19 g, 61.4 mmol) and 2-bromo-5-(difluoromethoxy)-3-methylpyrimidine (INT_008, 20.4 g, 85.7 mmol) in 1,4-dioxane (200 mL). The resulting mixture was stirred at 120°C for 16 hours. The reaction mixture was filtered and washed with SiO (200 mL). The filtrate was diluted with water (50 mL) and extracted with SiO (3 × 100 mL). The combined organic compounds were washed with brine (50 mL) and evaporated to dryness under vacuum. The residue was purified by recrystallization from MeCN (286 mL) to obtain the title compound as a solid (15.9 g, 55.5%). LCMS m / z = 467 [M+H] + ; 1H NMR(400 MHz, DMSO-d6) δ: 8.12 (d, 1H), 7.57(d, 1H), 7.47-7.40 (m, 1H), 7.26-7.01(m, 4H), 3.65(s, 2H), 2.34-2.25(m, 5H), 1.98-1.83(m, 10H), 1.52(br d, 2H).

[0220] EX-I-05: Synthesis of cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-3-(4-methyl-1H-pyrrolo[2,3-c]pyridine-5-yl)-1,3-diazaspiro[4.5]decan-2-one [ka]

[0221] To a 5 mL stirred solution of cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-3-(4-methyl-1-{[2-(trimethylsilyl)ethoxy]methyl}-1H-pyrrolo[2,3-c]pyridin-5-yl)-1,3-diazaspiro[4.5]decan-2-one (INT_015, 0.2 g, 0.35 mmol) in DCM, TFA (3 ml) was added at 0°C, and the reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure, and MeOH (5 ml), followed by aqueous NH3 solution (25%, 4 ml), was added, and the mixture was stirred for a further 1 hour. The mixture was concentrated, and the residue was purified by preparative HPLC to obtain the title compound as a solid (0.09 g, 58%). LCMS m / z = 440 [M+H] + ; 1 H NMR(400 MHz, DMSO-d6, 100 ℃) δ: 11.19(bs, 1H), 8.41(s, 1H), 7.50(d, 1H), 6.97-7.02(m, 3H), 6.69(s, 1H), 6.50(d, 1H), 3.64(s, 2H), 2.30-2.73(m, 5H), 2.06(s, 6H), 1.85-2.01(m, 4H), 1.55-1.60(m, 2H).

[0222] EX-I-06: Synthesis of cis-8-(3,5-difluorophenyl)-3-(5-(2-hydroxypropan-2-yl)-3-methylpyridine-2-yl)-8-(methylamino)-1,3-diazaspiro[4.5]decan-2-one [ka]

[0223] To a solution of 2-(6-chloro-5-methylpyridine-3-yl)propan-2-ol (INT_007, 1.84 g, 9.8 mmol) in 1,4-dioxane (30 mL), cis-8-(3,5-difluorophenyl)-8-(methylamino)-1,3-diazaspiro[4.5]decan-2-one (INT_018, 2.4 g, 8.2 mmol), xanthophos (0.71 g, 1.2 mmol), Cs2CO3 (7.9 g, 2.5 mmol), and Pd2(dba)3 (1.1 g, 1.2 mmol) were added at room temperature under an argon atmosphere. The resulting reaction mixture was stirred at 120 °C for 18 hours. The reaction mixture was cooled to room temperature, filtered through a Celite pad, and concentrated under vacuum. The residue was purified by preparative HPLC to obtain the title compound as a colorless solid (2.0 g, 58%). The solid was dissolved in MeCN (45 mL) at room temperature and stirred at 90°C for 30 minutes, followed by 60°C for 30 minutes. The solution was cooled to room temperature and stabilized for 18 hours. After recovering the substance by filtration, the title compound was obtained as a solid (1.90 g, 95%). LCMS m / z = 445.4 [M+H] + ; 1 H NMR(400 MHz, DMSO-d6, 25 ℃) δ: 8.29 (d, 1H), 7.67(d, 1H), 7.18-7.15(m, 2H), 7.04-6.99(m, 2H), 5.14(s, 1H), 3.76(s, 2H), 2.29-2.27(m, 1H), 2.23(s, 3H), 2.06-2.00(m, 2H), 1.89(d, 3H), 1.82-1.70(m, 4H), 1.60-1.57(m, 2H), 1.43(s, 6H).

[0224] EX-I-07: Synthesis of cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-3-(3-methyl-5-(trifluoromethoxy)pyridine-2-yl)-1,3-diazaspiro[4.5]decan-2-one [ka]

[0225] To a solution of cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-1,3-diazaspiro[4.5]decan-2-one (INT_004, 1.2 g, 3.9 mmol) in 1,4-dioxane (20 mL), 2-bromo-3-methyl-5-(trifluoromethoxy)pyridine (1.19 g, 4.66 mmol) and K2CO3 (1.6 g, 11.65 mmol) were added in a sealed tube under an argon atmosphere. Trans-N,N'-dimethylcyclohexane-1,2-diamine (0.12 mL, 0.78 mmol) and CuI (0.074 g, 0.388 mmol) were added, and the resulting mixture was stirred at 130 °C for 16 hours. The reaction mixture was cooled to room temperature and filtered through a Celite pad. The filtrate was concentrated under reduced pressure, and the residue was diluted with 10% MeOH-CH2Cl2 (150 ml) and washed with water (2 × 50 mL) and saturated NaCl solution (50 mL). The combined organic layers were dried on anhydrous Na2SO4 and concentrated under reduced pressure. The crude product was purified by combiflash chromatography (with 2-4% MeOH in DCM as the eluent) followed by preparative HPLC to obtain the title compound. This was crystallized from MeCN as described for EX-I-04 to obtain the title compound as a solid (1.15 g, 61%). LCMS m / z = 485.3 [M+H] + ; 1 H NMR(400 MHz, DMSO-d6, 100 ℃) δ: 8.24(s, 1H), 7.69(s, 1H), 7.20(s, 1H), 6.98-7.01(m, 3H), 3.69(s, 2H), 2.29-2.34(m, 5H), 2.05(s, 6H), 1.83-1.99(m, 4H), 1.53-1.58(m, 2H).

[0226] EX-I-08: Synthesis of cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-3-(4-methyl-6-(trifluoromethoxy)pyridazin-3-yl)-1,3-diazaspiro[4.5]decane-2-one hydrochloride [ka]

[0227] The free base of the title compound (0.17 g, 30%) was prepared similarly to EX-I-06 from INT_004 (0.36 g, 1.17 mmol) and 3-bromo-4-methyl-6-(trifluoromethoxy)pyridazine (0.3 g, 1.40 mmol). A suspension of the product (0.1 g, 0.21 mmol) in Et2O (5 mL) was treated with HCl (2 M in Et2O, 0.51 mL, 1.03 mmol), and the solution was gently warmed. The solvent was removed under reduced pressure to obtain the title compound as a solid (0.07 g, 65%). LCMS m / z = 486.5 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6, 25 ℃) δ: 10.1(s, 1H), 8.21(s, 1H), 7.68(s, 1H), 7.46-7.52(m, 3H), 3.71(s, 2H), 2.69(d, 2H), 2.59-2.60(m, 5H), 2.35(s, 3H), 2.23-2.30(m, 2H), 1.93(d, 2H), 1.39(t, 2H).

[0228] EX-I-09: Synthesis of cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-3-(5-(3-hydroxy-3-methylcyclobutoxy)-3-methylpyridine-2-yl)-1,3-diazaspiro[4.5]decan-2-one [ka]

[0229] The title compound (33 mg, 10%) was prepared similarly to EX-I-01 from INT_004 (0.21 g, 0.78 mmol) and 3-((6-bromo-5-methylpyridine-3-yl)oxy)-1-methylcyclobutan-1-ol (0.2 g, 0.65 mmol). LCMS m / z = 501.2 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6, 25 ℃) δ: 7.81(d, 1H), 7.24(br s, 1H), 7.17(d, 1H), 7.14-7.10(m, 1H), 7.04(s, 1H), 7.02(s, 1H), 5.14(s, 1H), 4.44-4.37(m, 1H), 3.57(s, 2H), 2.55-2.52(m, 2H), 2.32-2.29(m, 2H), 2.19(s, 3H), 2.08-2.03(m, 2H), 1.96(s, 6H), 1.88-1.86(m, 4H), 1.48-1.46(m, 2H), 1.25(s, 3H).

[0230] EX-I-10: Synthesis of cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-3-(3-methyl-5-(methylsulfonyl)pyridine-2-yl)-1,3-diazaspiro[4.5]decan-2-one [ka]

[0231] The title compound (0.33 g, 21%) was prepared similarly to EX-I-06 from INT_004 (1.0 g, 3.23 mmol) and 2-chloro-3-methyl-5-(methylsulfonyl)pyridine (1.0 g, 4.85 mmol). LCMS m / z = 479.2 [M+H] + ; 1H NMR(400 MHz, DMSO-d6, 25 ℃) δ: 8.65(d, 1H), 8.14(d, 1H), 7.72(br s, 1H), 7.13(br t, 1H), 7.04(br d, 2H), 3.75(s, 2H), 3.27(s, 3H), 2.34(s, 5H), 1.96(s, 6H), 1.94 - 1.79(m, 4H), 1.52(br s, 2H).

[0232] EX-I-11: Synthesis of cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-3-(5-(ethylsulfonyl)-3-methylpyridine-2-yl)-1,3-diazaspiro[4.5]decan-2-one [ka]

[0233] The title compound (0.15 g, 46%) was prepared similarly to EX-I-07 from INT_004 (0.2 g, 0.65 mmol) and 2-bromo-5-(ethylsulfonyl)-3-methylpyridine (0.17 g, 0.65 mmol). LCMS m / z = 493.3 [M+H] + ; 1 H NMR(400 MHz, DMSO-d6, 25 ℃) δ: 8.60(d, 1H), 8.09(d, 1H), 7.72(s, 1H), 7.12(t, 1H), 7.04(d, 2H), 3.76 (s, 2H), 3.35-3.37(m, 2H), 2.30-2.40(m, 5H), 1.79-1.96(m, 11H), 1.52-1.54(m, 2H), 1.11(t, 3H).

[0234] EX-I-12: Synthesis of cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-3-(2-methyl-6-(trifluoromethyl)pyridine-3-yl)-1,3-diazaspiro[4.5]decan-2-one [ka]

[0235] The title compound (95 mg, 44%) was prepared similarly to EX-I-01 from INT_004 (0.14 g, 0.45 mmol) and 3-bromo-2-methyl-6-(trifluoromethyl)pyridine (119 mg, 0.50 mmol). LCMS m / z = 469.4 [M+H] + ; 1 H NMR(400 MHz, DMSO-d6, 25 ℃) δ: 7.88(d, 1H), 7.73(d, 1H), 7.52(brs, 1H), 7.15-7.10(s, 1H), 7.06-7.04(m, 2H), 3.60(s, 2H), 2.46(s, 3H), 2.40-2.30(m, 2H), 1.97-1.80(m, 10H), 1.53-1.49(m, 2H).

[0236] EX-I-13: Synthesis of cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-3-(2,6-dimethylpyridine-3-yl)-1,3-diazaspiro[4.5]decan-2-one [ka]

[0237] The title compound (40 mg, 15%) was prepared similarly to EX-I-07 from INT_004 (0.20 g, 0.65 mmol) and 3-bromo-2,6-dimethylpyridine (144 mg, 0.78 mmol). LCMS m / z = 415.4 [M+H] + ; 1 H NMR(400 MHz, DMSO-d6, 100 ℃) δ: 7.43(d, 1H), 7.0-7.04(m, 4H), 6.89(s, 1H), 3.45(s, 2H), 2.41(s, 3H), 2.18-2.34(m, 5H), 2.06(s, 6H), 1.85-1.98(m, 4H), 1.52-1.56(m, 2H).

[0238] EX-I-14: Synthesis of cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-3-(2-methyl-6-(trifluoromethoxy)pyridine-3-yl)-1,3-diazaspiro[4.5]decane-2-one hydrochloride [ka]

[0239] The free base of the title compound (0.11 g, 17%) was prepared similarly to EX-I-06 from INT_004 (0.40 g, 1.29 mmol) and 3-bromo-2-methyl-6-(trifluoromethoxy)pyridine (0.40 g, 1.55 mmol). A suspension of the product (0.11 g, 0.22 mmol) in H2O (5 mL) was treated with HCl (1 M in H2O, 0.32 mL, 0.22 mmol), and the mixture was gently warmed to obtain a solution. The solvent was removed under reduced pressure to obtain the title compound as a solid (0.07 g, 65%). LCMS m / z = 10.49(s, 1H), 7.84(d, 1H), 7.73(s, 1H), 7.54(d, 2H), 7.44(t, 1H), 7.14(d, 1H), 3.39(s, 2H), 2.70(d, 2H), 2.58(d, 6H), 2.28-2.33(m, 5H), 1.91(d, 2H), 1.33(t, 2H).

[0240] EX-I-15: Synthesis of cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-3-(4-methyl-2-(oxetan-3-yloxy)pyrimidine-5-yl)-1,3-diazaspiro[4.5]decan-2-one [ka]

[0241] The title compound (88 mg, 29%) was prepared similarly to EX-I-07 from INT_004 (0.20 g, 0.65 mmol) and 5-bromo-4-methyl-2-(oxetane-3-yloxy)pyrimidine (191 mg, 0.78 mmol). LCMS m / z = 474.2 [M+H]+ ; 1 H NMR(400 MHz, DMSO-d6, 25 ℃) δ: 8.38(s, 1H), 7.36-7.43(m, 1H), 7.03-7.12(m, 3H), 5.49-5.52(m, 1H), 4.84-4.88(m, 2H), 4.53-4.56(m, 2H), 3.47(s, 2H), 2.29-2.37(m, 5H), 1.80-1.96(m, 10H),1.36-1.48(m, 2H).

[0242] EX-I-16: Synthesis of cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-3-(2-(2-methoxyethoxy)-4-methylpyrimidine-5-yl)-1,3-diazaspiro[4.5]decan-2-one [ka]

[0243] The title compound (65 mg, 27%) was prepared similarly to EX-I-01 from INT_004 (0.20 g, 0.64 mmol) and 5-bromo-2-(2-methoxyethoxy)-4-methylpyrimidine (322 mg, 1.29 mmol). LCMS m / z = 476.2 [M+H] + ; 1 H NMR(400 MHz, DMSO-d6, 25 ℃) δ: 8.37(s, 1H), 7.34(br s, 1H), 7.15-7.09(m, 2H), 7.06-7.03(m, 2H), 4.39-4.36(m, 2H), 3.65-3.62(m, 2H), 3.47-3.45(br s, 2H), 3.28(s, 3H), 2.33-2.29(m, 5H), 1.96-1.80(m, 9H), 1.50(br s, 2H).

[0244] EX-I-17: Synthesis of cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-3-(2-methyl-6-(methylsulfonyl)pyridine-3-yl)-1,3-diazaspiro[4.5]decan-2-one [ka]

[0245] The title compound (0.13 g, 56%) was prepared similarly to EX-I-06 from INT_004 (0.15 g, 0.49 mmol) and 3-bromo-2-methyl-6-(methylsulfonyl)pyridine (0.18 g, 0.73 mmol). LCMS m / z = 479.2 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6, 25 ℃) δ: 7.93(d, 1H), 7.87(d, 1H), 7.57(br s, 1H), 7.16-7.08(m, 1H), 7.07-7.02(m, 2H), 3.61(s, 2H), 3.24(s, 3H), 2.49(s, 3H), 2.46-2.28(m, 2H), 1.96-1.70(m, 10H), 1.60-1.48(m, 2H).

[0246] EX-I-18: Synthesis of cis-3-(2-cyclopropoxy-4-methylpyrimidine-5-yl)-8-(3,5-difluorophenyl)-8-(dimethylamino)-1,3-diazaspiro[4.5]decan-2-one [ka]

[0247] A mixture of 5-bromo-2-cyclopropoxy-4-methylpyrimidine (INT_019, 19 g, 82.9 mmol), cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-1,3-diazaspiro[4.5]decan-2-one (INT_004, 23.3 g, 75.4 mmol), and K3PO4 (48 g, 226 mmol) in 1,4-dioxane (1 L) was stirred at 25°C for 30 minutes. CuI (14.4 g, 75.4 mmol) and DMEDA (13.3 g, 151 mmol) were added, and the mixture was stirred at 120°C for 16 hours. The solids were removed by filtration and washed with SiO2 (100 mL). The filtered material was extracted with ÃO (500 mL x 3), the combined organic matter was evaporated under reduced pressure and dried, and the residue was purified by column chromatography (SiO2, 9-100% MeOH / DCM). The residue was further purified by grinding with MeCN (260 ml) at 90°C for 30 minutes, cooled to 50°C, and then MeCN (13 ml) was added at 50°C. The mixture was stirred at 90°C for 30 minutes, filtered, cooled to 50°C, stirred at 90°C for 10 minutes, and then stirred at 25°C for 16 hours. The solid was collected and dried under reduced pressure. The obtained solid was ground with MTBE at 25°C for 2 hours, the solid was collected by filtration, and the solid was dried under reduced pressure. The solid was stirred with MeCN (260 mL) at 90°C for 30 minutes, cooled to 50°C, diluted with MeCN (13.0 mL) at 50°C, and stirred at 90°C for 30 minutes. The hot solution was filtered, cooled to 50°C, heated at 90°C for 10 minutes, and then stirred at 25°C for 16 hours. The solid was recovered by filtration and dried under reduced pressure to obtain the title compound as a solid (18.1 g, 52%). LCMS m / z = 458 [M+H] + ; 1 H NMR(400 MHz, DMSO-d6) δ: 8.39(s, 1H), 7.36(br s, 1H), 7.12(br t, 1H), 7.05(br d, 2H), 4.25(tt, 1H), 3.48(s, 2H), 2.42-2.22(m, 5H), 2.03-1.69(m, 10H), 1.57-1.42(m, 2H), 0.78-0.72(m, 2H), 0.71-0.65(m, 2H).

[0248] EX-I-19: Synthesis of cis-8-(3,5-difluorophenyl)-3-(1,3-dimethyl-2-oxo-1,2-dihydropyridine-4-yl)-8-(dimethylamino)-1,3-diazaspiro[4.5]decan-2-one [ka]

[0249] K3PO4 (13.4 g, 63.1 mmol) was added at room temperature to an argon-degassed solution of cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-1,3-diazaspiro[4.5]decan-2-one (INT_004, 6.5 g, 21.0 mmol) and 4-bromo-1,3-dimethylpyridine-2(1H)-one (INT_020, 5.1 g, 25.2 mmol) in dioxane (200 mL). CuI (4.01 g, 21 mmol) and DMEDA (2.26 mL, 21.0 mmol) were added sequentially to the resulting suspension. The reaction mixture was degassed for 10 minutes and then heated at 120 °C for 18 hours. The reaction mixture was cooled to room temperature, filtered through a Celite pad, and washed with siRNA (300 mL). The filtrate was diluted with water (800 mL), and the organic layer was separated. The combined organic matter was dried (Na2SO4) and evaporated to dryness under vacuum to obtain the title compound as a white solid (9.8 g, crude). The solid in acetonitrile (350 mL) was heated to 90°C for 2.5 hours. The resulting clear solution was then stirred at 60°C for 30 minutes, then slowly cooled to room temperature and stirred for 18 hours. The solid was recovered by filtration and dried under reduced pressure to obtain the title compound as a solid (5.5 g). The filtrate was evaporated under reduced pressure to obtain an additional 1.7 g of the title compound as a solid. Total yield: 7.2 g, 80%). LCMS m / z = 431 [M+H] + ; 1H NMR(500 MHz, DMSO-d6) δ: 7.48(d, 1H), 7.37(br s, 1H), 7.14-7.10(m, 1H), 7.05-7.03(m, 2H), 6.18(d, 1H), 3.46(s, 2H), 3.38(s, 3H), 2.40-2.22(m, 2H), 1.95-1.70(m, 13H), 1.57-1.46(m, 2H).

[0250] EX-I-20: Synthesis of cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-3-(5-(2-hydroxypropan-2-yl)-3-methylpyrazine-2-yl)-1,3-diazaspiro[4.5]decan-2-one [ka]

[0251] The title compound (1.4 g, 78%) was prepared similarly to EX-I-06 from INT_004 (1.2 g, 3.9 mmol) and 2-(5-chloro-6-methylpyrazine-2-yl)propan-2-ol (INT_021, 0.87 g, 4.7 mmol). LCMS m / z = 460 [M+H] + ; 1 H NMR(400 MHz, DMSO-d6) δ: 8.45(s, 1H), 7.54(brs, 1H), 7.12(t, 1H), 7.04(d, 2H), 5.32(s, 1H), 3.68(s, 2H), 2.43(s, 3H), 2.32(m, 2H), 1.96(s, 6H), 1.88-1.93(m. 4H), 1.51(m, 2H), 1.43(s, 6H).

[0252] EX-I-21: Synthesis of cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-3-(2-(3-fluorooxetan-3-yl)-4-methylpyrimidine-5-yl)-1,3-diazaspiro[4.5]decan-2-one [ka]

[0253] The title compound (130 mg, 45%) was prepared similarly to EX-I-01 from INT_004 (0.21 g, 0.67 mmol) and 5-bromo-2-(3-fluorooxetan-3-yl)-4-methylpyrimidine (INT_024, 150 mg, 0.61 mmol). LCMS m / z = 475 [M+H] + ; 1 H NMR(500 MHz, DMSO-d6) δ: 8.73(s, 1H), 7.56(brs, 1H), 7.15-7.11(m, 1H), 7.07-7.05(m, 2H), 5.09-5.04(m, 2H), 4.95-4.89(m, 2H), 3.62(s, 2H), 2.45(s, 3H), 2.37-2.32(m, 2H), 1.97-1.84(m, 10H), 1.54-1.48(m, 2H).

[0254] EX-I-22: Synthesis of cis-8-(3,5-difluorophenyl-8-(dimethylamino)-3-(2-(2-hydroxypropan-2-yl)-4-methylpyrimidine-5-yl)-1,3-diazaspiro[4.5]decan-2-one [ka]

[0255] The title compound (130 mg, 45%) was prepared similarly to EX-I-01 from INT_004 (0.26 g, 0.85 mmol) and 2-(5-bromo-4-methylpyrimidine-2-yl)propan-2-ol (INT_027, 236 mg, 1.02 mmol). LCMS m / z = 460 [M+H] + ; 1H NMR(400 MHz, DMSO-d6) δ: 8.55(s, 1H), 7.15(s, 1H), 7.02-7.00(m, 3H), 4.63(s, 1H), 3.57(s, 2H), 2.43(s, 3H), 2.36-2.31(m, 2H), 2.07(s, 6H), 2.01-1.95(m, 2H), 1.88-1.85(m, 2H), 1.60-1.55(m, 2H), 1.50(s, 6H).

[0256] EX-I-23: Synthesis of cis-3-(5-(1,1-difluoroethoxy)-1-methyl-6-oxo-1,6-dihydropyridine-2-yl)-8-(3,5-difluorophenyl)-8-(dimethylamino)-1,3-diazaspiro[4.5]decan-2-one [ka]

[0257] 6-Chloro-3-(1,1-difluoroethoxy)-1-methylpyridine-2(1H)-one (INT_029, 1.62 g, 6.5 mmol) and Cs2CO3 (5.7 g, 17.6 mmol) were added to a solution of cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-1,3-diazaspiro[4.5]decan-2-one (INT_004, 1.8 g, 5.9 mmol) in dioxane (100 mL). Xanthophos (509 g, 0.9 mmol) and Pd2(dba)3 (805 mg, 0.9 mmol) were added, and the solution was stirred at 120 °C for 16 hours under a nitrogen atmosphere. The reaction product was quenched by the addition of H2O (60 mL), and the mixture was extracted with ELISA (3 × 50 mL). The combined organic layers were dried over anhydrous Na2SO4 and concentrated under vacuum. The residue was purified by column chromatography (SiO2, 1-100% siRNA / PE) to obtain the title compound (1.8 g, 62%) as a solid. LCMS m / z = 497 [M+H] + ; 1H NMR(400 MHz, DMSO-d6) δ: 1.52(br d, 2 H) 1.71 - 2.02(m, 13 H) 2.21 - 2.39(m, 2 H) 3.38(s, 3 H) 3.55(s, 2 H) 6.20(d, 1 H) 7.01 - 7.08(m, 2 H) 7.13(br t, 1 H) 7.36(d, 1 H) 7.73(br s, 1 H).

[0258] EX-I-24: Synthesis of 8-(3,5-difluorophenyl)-8-(dimethylamino)-3-(3-methyl-5-((S)-1,1,1-trifluoro-2-hydroxypropan-2-yl)-pyridine-2-yl)-1,3-diazaspiro[4.5]decan-2-one (enantiomer-2) [ka]

[0259] In a sealed tube under an argon atmosphere, xanthophos (70 mg, 0.12 mmol) and Pd2(dba)3 (110 mg, 0.12 mmol) were added at room temperature to a stirred solution of cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-1,3-diazaspiro[4.5]decan-2-one (INT_004, 0.25 g, 0.81 mmol), 2-(6-chloro-5-methylpyridine-3-yl)-1,1,1-trifluoropropan-2-ol (INT_031, 0.232 g, 0.970 mmol), and Cs2CO3 (0.788 g, 2.427 mmol) in 1,4-dioxane (20 mL). The reaction mixture was purged with argon and degassed for a further 10 minutes. The resulting reaction mixture was heated to 120 °C and stirred at that temperature for 16 hours. The reaction mixture was cooled to room temperature, filtered through a Celite pad, and washed with ethyl acetate (3 × 100 mL). The filtrate was concentrated under vacuum, and the residue was purified by reverse-phase column chromatography to obtain the racemic compound as a solid (400 mg, 96%). The racemic material was separated by chiral SFC (Chiralpak IC5 μM, 80% CO2 20% 0.5%-Et2NH / MeOH 3 mL / min) to obtain a single, secondary-eluted enantiomer as a solid (90 mg). LCMS m / z = 413 [M+H] + ; 1 H NMR(400 MHz, DMSO-d6) δ: 8.36(d, 1H), 7.79 (d, 1H), 7.43(brs, 1H), 7.15-7.09(m, 1H), 7.05-7.03(m, 2H), 6.74(s, 1H), 3.67(s, 2H), 2.33-2.26(m, 5H), 1.96-1.87(m, 10H), 1.69(s, 3H), 1.52-1.48(m, 2H).

[0260] EX-I-25: Synthesis of cis-3-(5-cyclopropoxy-3-methylpyridine-2-yl)-8-(3,5-difluorophenyl)-8-(methylamino)-1,3-diazaspiro[4.5]decan-2-one [ka]

[0261] The title compound was prepared from cis-8-(3,5-difluorophenyl)-8-(methylamino)-1,3-diazaspiro[4.5]decan-2-one (INT_018) and 2-bromo-5-cyclopropoxy-3-methylpyridine (INT_005) using a method similar to that described for EX-I-01. The solid was crystallized from MeCN as described for EX-I-06 to give the title compound as a solid (2.39 g, 64%). LCMS m / z = 443.2 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6, 25 °C) δ: 7.99 (d, 1H), 7.38 (d, 1H), 7.17 - 7.14 (m, 2H), 7.04 - 6.98 (m, 2H), 3.94 - 3.90 (m, 1H), 3.72 (s, 2H), 2.32 - 2.23 (m, 4H), 2.06 - 2.00 (m, 2H), 1.90 (d, 3H), 1.82 - 1.57 (m, 6H), 0.81 - 0.77 (m, 2H), 0.68 - 0.64 (m, 2H).

[0262] EX-I-26: Synthesis of cis-3-(5-(2,2-difluoromethoxy)-3-methylpyridin-2-yl)-8-(3,5-difluorophenyl)-8-(dimethylamino)-1,3-diazaspiro[4.5]decan-2-one

Chemical Structure

[0263] The title compound (54 mg, 26%) was prepared from INT_004 (0.16 g, 0.52 mmol) and 2-chloro-5-(2,2-difluoroethoxy)-3-methylpyridine (INT_032, 0.12 g, 0.58 mmol) similar to EX-I-06. LCMS m / z = 479.3 [M-H] - , 1H NMR(400 MHz, DMSO-d6) δ: 7.98(d, 1H), 7.39(d, 1H), 7.28(bs, 1H), 7.12(t, 1H), 7.04-7.02(m, 2H), 6.38(tt, 1H), 4.37(td, 2H), 3.59(s, 2H), 2.32-2.22(m, 5H), 1.96-1.86(m, 10H), 1.51-1.49(m, 2H).

[0264] SC_027: Synthesis of cis-3-(6-(2,2-difluoroethoxy)-4,5-dimethylpyridazin-3-yl)-8-(3,5-difluorophenyl)-8-(dimethylamino)-1,3-diazaspiro[4.5]decan-2-one [ka]

[0265] The title compound was prepared as a solid (135 mg, 40%) from cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-1,3-diazaspiro[4.5]decan-2-one (INT_004, 0.20 g, 0.65 mmol) and 3-chloro-6-(2,2-difluoroethoxy)-4,5-dimethylpyridazine (INT_033, 0.17 g, 0.78 mmol) using a method similar to that described for EX-I-06. LCMS m / z = 494.4 [MH] - , 1 H NMR(400 MHz, DMSO-d6) δ: 7.56(br, 1H), 7.15-7.04(m, 3H), 6.44(tt, 1H), 4.69(td, 2H), 3.71(s, 2H), 2.32(m, 2H), 2.16(s, 6H), 1.96-1.89(m, 10H), 1.52(m, 2H).

[0266] Synthesis of EX-I-28: cis-3-(5-(Difluoromethoxy)-3-methylpyridin-2-yl)-8-(3,5-difluorophenyl)-8-(dimethylamino)-1,3-diazaspiro[4.5]decane-2,4-dione

Chemical formula

[0267] Cu2O (98 mg, 0.52 mmol) and DMEDA (46 mg, 0.52 mmol) were added to a stirred solution in a microwave vial at room temperature in DMA (10 mL) of cis-8-(3,5-difluorophenyl)-8-(dimethylamino)-1,3-diazaspiro[4.5]decane-2,4-dione (INT_003, 0.21 g, 0.65 mmol) and 5-(difluoromethoxy)-2-iodo-3-methylpyridine (INT_034, 184 mg, 0.65 mmol). The reaction mixture was stirred in the microwave at 180 °C for 2 hours, then cooled to room temperature, filtered through a Celite pad, and washed with dichloromethane (2 × 20 mL). The filtrate was concentrated in vacuo, and the crude product was purified by preparative HPLC to give the title compound (35 mg, 11%) as a solid. LCMS m / z = 479.3 [M-H] - , 1 1H NMR (400 MHz, DMSO-d6) δ: 9.09 (br, 1H), 8.31 (d, 1H), 7.76 (d, 1H), 7.36 (t, 1H), 7.20 - 7.14 (m, 1H), 7.11 - 7.08 (m, 2H), 2.51 (s, 2H), 2.14 (s, 3H), 2.07 - 2.02 (m, 2H), 1.95 (s, 6H), 1.86 - 1.78 (m, 2H), 1.65 - 1.60 (m, 2H).

[0268] Pharmacological studies

[0269] Functional studies of human mu-opioid receptors (hMOP), human kappa-opioid receptors (hKOP), human delta-opioid receptors (hDOP), and human nociceptin / olphanin FQ peptide receptors (hNOP).

[0270] Human NOP binding assay (Ardati, A., et al. (1997), Mol. Pharmacol., 51: 816-824).

[0271] Human nociceptin (hNOP) receptor binding assays were performed as filtration-based radioactive agonist binding assays. Cell membrane homogenates of transfected Chem-1 cells (5 μg) were incubated with 0.1 nM [3H]nociceptin in a buffer containing 50 mM Tris-HCl (pH 7.4), 5 mM MgCl2, and 0.1% BSA in a 96-well plate with a final volume of 200 μl, in the absence or presence of the test compound, at 22°C for 60 minutes. Nonspecific binding was determined in the presence of 1 μM nociceptin.

[0272] The test compounds were added to a 100-fold concentrated solvent solution, with a final assay DMSO concentration of up to 1%, which also served as a vehicle control. After incubation, the samples were rapidly filtered under vacuum through a glass fiber filter (GF / B, Packard, pre-soaked with 0.3% PEI) and washed several times with ice-cold 50 mM Tris-HCl using a 96-sample cell harvester (Unifilter, Packard). The filters were dried, and radioactivity was counted using a scintillation cocktail (Microscint 0, Packard) and a scintillation counter (Topcount, Packard).

[0273] The results are expressed as the percentage of inhibition of specific binding of the control radioligand. The maximum half-dose inhibitory concentration (IC50), which reflects 50% substitution of [3H]nociceptin-specific receptor binding, is calculated by nonlinear regression analysis, and the Ki value is calculated using the Cheng-Prusoff equation (Cheng Y. and Prusoff WH (1973), Biochem. Pharmacol. 22:3099-3108).

[0274] Human DOP binding assay (Simonin, F. et al. (1994), Mol. Pharmacol., 46:1015-1021)

[0275] Human δ2-opioid (hDOP) receptor binding assays were performed as filtration-based radioactive agonist binding assays. Cell membrane homogenates of transfected Chem-1 cells (1 μg) were incubated in a 96-well plate with a final volume of 200 μl in buffer containing 50 mM Tris-HCl (pH 7.4) and 5 mM MgCl2, in the absence or presence of the test compound, with 0.5 nM [3H]DADLE at 22°C for 60 minutes. Nonspecific binding was determined in the presence of 10 μM naltrexone.

[0276] The test compounds were added to a 100-fold concentrated solvent solution, and the final assay DMSO concentration was up to 1%, which also served as a vehicle control for each compound.

[0277] After incubation, the samples were rapidly filtered under vacuum through a glass fiber filter (GF / B, Packard, pre-soaked with 0.3% PEI) and washed several times with ice-cold 50 mM Tris-HCl using a 96-sample cell harvester (Unifilter, Packard). The filters were dried, and radioactivity was counted using a scintillation cocktail (Microscint 0, Packard) and a scintillation counter (Topcount, Packard).

[0278] The results are expressed as the percentage of inhibition of specific binding of the control radioligand. The maximum half-dose inhibitory concentration (IC50), which reflects 50% substitution of [3H]DADLE-specific receptor binding, is calculated by nonlinear regression analysis, and the Ki value is calculated using the Chen-Prusov equation.

[0279] Human MOP binding

[0280] Human μ-opioid (hMOP) receptor binding assays were performed as filtration-based radioactive agonist binding assays. Transfected CHO cells (10 μg) cell membranes were incubated in a 96-well plate with 100 μl final volume in a buffer containing 50 mM Tris, 5 mM MgCl2, and 10 μg / ml saponin, with 0.7 nM [3H] DAMGO for 60 minutes, either in the absence or in the presence of the test compound. Nonspecific binding was determined in the presence of 10 μM DAMGO.

[0281] After incubation, the samples were rapidly filtered through a filter plate (GF / C). The filters were washed six times with 0.5 ml of ice-cold wash buffer, and 50 μl of Microscint 20 (Packard) was added to each well. The plates were incubated on an orbital shaker for 15 minutes, and then counted using TopCount® at a rate of 30 seconds per well.

[0282] The results are expressed as the percentage of inhibition of specific binding of the control radioligand. The maximum half-dose inhibitory concentration (IC50), which reflects 50% substitution of [3H]DAMGO-specific receptor binding, is calculated by nonlinear regression analysis, and the Ki value is calculated using the Chen-Prusov equation.

[0283] Human KOP binding

[0284] Human κ-opioid (hKOP) receptor assays were performed as filtration-based radioactive agonist binding assays. Transfected CHO cells (15 μg) cell membranes were incubated in a 96-well plate with 100 μl final volume in a buffer containing 50 mM Tris, 5 mM MgCl2, and 10 μg / ml saponin, with 0.9 nM [3H]U-69593 in the absence or presence of the test compound, at 25°C for 60 minutes. Nonspecific binding was determined in the presence of 10 μM U-50488.

[0285] After incubation, the samples were rapidly filtered through a filter plate (GF / C). The filters were washed six times with 0.5 ml of ice-cold wash buffer, and 50 μl of Microscint 20 (Packard) was added to each well. The plates were incubated on an orbital shaker for 15 minutes, and then counted using TopCount® at a rate of 30 seconds per well.

[0286] The results are expressed as the percentage of inhibition of specific binding of the control radioligand. The maximum half-mass inhibitory concentration (IC50), which reflects 50% substitution of [3H]U-69593-specific receptor binding, is calculated by nonlinear regression analysis, and the Ki value is calculated using the Chen-Prusov equation.

[0287] The results are summarized in the table below. [Table 1]

[0288] Comparative example: [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] Table 2-5 Table 2-6

Claims

1. Compounds of general formula (I), 【Chemistry 1】 During the ceremony, A1 represents N or CR1, A2 represents N or C, A3 represents N or CH, A4 represents N or CH, A5 represents N or C, (However, one or two of A1, A2, A3, A4, and A5 represent N, and the remaining three or four of A1, A2, A3, A4, and A5 do not represent N.) R1 is -H, -CH ３ Or, it represents O, R2 is, -C １ -C ６ -Alkyl (linear or branched, saturated or unsaturated, unsubstituted, or substituted with one, two, three, or four substituents independently selected from -F and -OH); -C ３ -C ７ - Heterocycloalkyl (saturated or unsaturated, unsubstituted, or -F, -OH and -CH) ３ (Substituted with one, two, three, or four substituents independently selected from the original molecule); -S(O) ２ -C １ -C ４ -alkyl (linear, branched or cyclic, unsubstituted or substituted with one, two or three substituents -F); -O-C １ -C ６ -Alkyl (linear or branched, saturated or unsaturated, unsubstituted, or -F, -OH and -OCH) ３ (Substituted with one, two, three, or four substituents independently selected from the original molecule); -O-C ３ -C ８ -Cycloalkyl (saturated or unsaturated, unsubstituted, or -F, -OH and -CH) ３ (substituted with one, two, three, or four substituents independently selected from); or -O-C ３ -C ７ - Represents a heterocycloalkyl group (saturated or unsaturated, unsubstituted, or substituted with one, two, three, or four substituents independently selected from -F and -OH); Alternatively, R1 and R2 together with A1 and A2 form a ring, meaning -CH=CN-NH-. R3 is -H or -CH ３ This represents, Both R4 and R5 represent -H, or together they mean =O. or a physiologically acceptable salt thereof.

2. (i) one of A1, A2, A3, A4, and A5 represents N, while the remaining four of A1, A2, A3, A4, and A5 do not represent N; or (ii) The compound according to claim 1, wherein two of A1, A2, A3, A4, and A5 represent N, and the remaining three of A1, A2, A3, A4, and A5 do not represent N.

3. The compound according to claim 1 or 2, wherein A1 represents CR1, A2 represents C, A3 represents CH, A4 represents N; and A5 represents C.

4. The compound according to claim 1 or 2, wherein A1 represents N, A2 represents C, A3 represents CH, A4 represents CH; and A5 represents C.

5. The compound according to claim 1 or 2, wherein A1 represents CR1, A2 represents C, A3 represents N, A4 represents N; and A5 represents C.

6. The compound according to claim 1 or 2, wherein A1 represents N, A2 represents C, A3 represents N, A4 represents CH; and A5 represents C.

7. The compound according to claim 1 or 2, wherein A1 represents CR1 together with R1 which means =O, A2 represents N, A3 represents CH, A4 represents CH; and A5 represents C.

8. The compound according to claim 1 or 2, wherein A1 represents N, A2 represents C, A3 represents CH, A4 represents N; and A5 represents C.

9. The compound according to claim 1 or 2, wherein A1 represents CR1 together with R1 which means =O, A2 represents C, A3 represents CH, A4 represents CH; and A5 represents N.

10. If R1 is present, then -H or -CH ３ A compound according to any of the prior claims, representing the compound.

11. R2 is -CH ３ , -CF ３ , -C(CH ３ ) ２ OH, and -C(CH ３ ) (OH) CF ３ A compound selected from any of the prior claims.

12. The compound according to any one of claims 1 to 10, wherein R2 represents 3-fluorooxetan-3-yl.

13. R2 is -S(O) ２ CH ３ and -S(O) ２ CH ２ CH ３ A compound according to any one of claims 1 to 10, selected from the above.

14. R2 is -OCH ３ , -OCHF ２ , -OCH ２ CHF ２ , -OCF ２ CH ３ , -OCF ３ , -OCH ２ CH ２ OCH ３ A compound according to any one of claims 1 to 10, selected from -O-cyclopropyl and 3-hydroxy-3-methylcyclobutoxy.

15. The compound according to any one of claims 1 to 10, wherein R2 represents oxetane-3-yloxy.

16. The compound according to any one of claims 1 to 10, wherein R1 and R2 together with A1 and A2 form a ring, and the relationship is -CH=CN-NH-.

17. R3 is -CH ３ A compound according to any of the prior claims, representing the compound.

18. The compound according to any of the prior claims, wherein both R4 and R5 mean -H.

19. A compound selected from the group consisting of the following, according to any of the prior claims, Table 1-1 Table 1-2 Table 1-3 and its physiologically acceptable salts.

20. A pharmaceutical product comprising a compound described in any of the prior claims.

21. A compound according to any one of claims 1 to 13, for use in the treatment of pain.

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