Therapeutic Compounds and Methods

Abstract

The present disclosure provides a compound of formula I or a salt thereof (wherein R 1 , R 2 , R 4 , L 1 , L 2 , and A have any of the values described herein), and a composition comprising a compound of formula (I). The compound 5 is useful as a TRPV4 antagonist. TIFF2025522953000269.tif66157

Description

Technical Field

[0001] Priority This application claims priority to U.S. Provisional Application No. 63 / 359,715, filed on July 8, 2022, and U.S. Provisional Application No. 63 / 385,282, filed on November 29, 2022. The entire contents of each of these U.S. provisional patent applications are hereby incorporated by reference.

[0002] Field The present disclosure relates to pharmaceutical compositions that act as antagonists of both wild-type TRPV4 channels and TRPV4 channels carrying activating mutations that cause disease.

Background Art

[0003] Background Transient receptor potential vanilloid 4, TRPV4, is a member of the transient receptor potential (TRP) superfamily (VenKatachlam and CraigMontell 2007 Annual Reviews Biochemistry). The TRPV4 gene encodes a non-selective cation channel that is highly permeable to calcium. TRPV4 is activated by a variety of different stimuli, including heat, mechanical stress, and chemicals, including arachidonic acid metabolites. TRPV4 is expressed in a number of different tissues, including, among others, the brain, bladder, skin, heart, lung, and musculoskeletal tissues.

[0004] The TRPV4 channel is widely expressed in a variety of human cell types. In particular, TRPV4 is expressed in epithelial and endothelial cells, fibroblasts, chondrocytes, neurons, and various inflammatory cells (Koivisto et al 2021 NRDD). TRPV4 has been directly involved in epithelial and endothelial barrier functions in relation to lung injury and has been further associated with respiratory diseases through TRPV4-induced ATP release (Koivisto). Genetic and pharmacological studies have linked TRPV4 as a therapeutic target for chronic cough, pulmonary edema, chronic obstructive pulmonary disease, and pulmonary fibrosis (Grace et al 2017, Pharmacology and Therapeutic). Genetic knockout of TRPV4 results in reduced osteoclast function and calcium regulation, which is important for bone homeostasis, suggesting the role of TRPV4 in osteoporosis and other joint diseases. In TRPV4 knockout mice, inflammatory hyperalgesia and mechanical pain are reduced (J. Neurosci.2004,18,4444-4452,Qu,et al.,2016 BioMed Research International), and various functional studies have demonstrated TRPV4 signaling in neuropathic pain. TRPV4 is also expressed in the urothelium and detrusor muscle of the bladder, and its activation causes muscle contraction (Birder et al 2007 J.Pharmacol. Exp. Ther.). Consistent with the role of TRPV4 in bladder-related conditions, inhibition of TRPV4 has been shown to improve bladder function in mouse and rat models of cyclophosphamide-induced cystitis (Prog. Biophys. Mol. Biol.2010, 1, 2-17). This provides evidence for TRPV4 as a therapeutic target in multiple diseases including respiratory diseases, joint diseases, pain, and bladder dysfunction.

[0005] Furthermore, activation of pathogenic mutations in TRPV4 has been shown to cause multiple severe Mendelian diseases, including a spectrum of skeletal dysplasias (Nishiura et al 2012 AJMG) and peripheral neuropathies (Landoure et al. Nature Genetics 2009). Mutations underlying all of these diseases appear to contribute to disease risk by increasing calcium influx into cells (Toft-Beterlsen and MacAulay 2021 Cells), suggesting a therapeutic effect of TRPV4 inhibitors. These observations suggest the benefits of inhibiting TRPV4 in genetic diseases due to activating mutations, in addition to multiple different common diseases caused by activation of the wild-type receptor.

[0006] GSK2798745 advanced to a Phase II clinical trial. Further exploration revealed circulating active metabolites (ACS Med. Chem. Lett. 2021,12,9,1498-1502), and the safety margin established by a 3-month canine toxicity study became lower, resulting in a 4.8-fold decrease in the maximum amount clinically administered. Dose (Am.J.Cardiovasc. Drugs 2019,19,335-342). GSK2798745 was unable to demonstrate efficacy across several human disease indications. Currently, there is a need for agents useful for antagonizing TRPV4. In particular, there is a need for agents having lower toxicity, improved potency, improved metabolic stability (e.g., against CYP3A4), lower levels of active circulating metabolites, and / or a higher safety margin. SUMMARY OF THE INVENTION

[0007] In one aspect, the present disclosure provides compounds useful for antagonizing TRPV4.

[0008] In some aspects, the present disclosure provides a compound of Formula I: TIFF2025522953000002.tif58165 or a salt thereof, wherein L 1 is CR aR b is, L 2 is CR c R d is, R a is H, halo, cyano, (C1-C5)alkyl, (C1-C5)alkoxy, (C3-C5)cyclopropyloxy, or (C3-C5)cycloalkyl, and any (C1-C5)alkyl, (C1-C5)alkoxy, (C3-C5)cyclopropyloxy, and (C3-C5)cycloalkyl may be substituted with one or more fluoros, R b is H, halo, cyano, (C1-C5)alkyl, (C1-C5)alkoxy, or (C3-C5)cycloalkyl, and any (C1-C5)alkyl, (C1-C5)alkoxy, and (C3-C5)cycloalkyl may be substituted with one or more fluoros, or R a and R b together are oxo(=O)methylene(=CH2), or R a and R b together with the atom to which they are attached form spiro(C3-C5)cycloalkyl, R c is H, halo, cyano, (C1-C5)alkyl, (C1-C5)alkoxy, (C3-C5)cyclopropyloxy, or (C3-C5)cycloalkyl, and any (C1-C5)alkyl, (C1-C5)alkoxy, (C3-C5)cyclopropyloxy, and (C3-C5)cycloalkyl may be substituted with one or more fluoros, R d is H, halo, cyano, (C1-C5)alkyl, (C1-C5)alkoxy, or (C3-C5)cycloalkyl, and any (C1-C5)alkyl, (C1-C5)alkoxy, and (C3-C5)cycloalkyl may be substituted with one or more fluoros, or R c and R 1 together with the atom to which they are attached form a fused cyclopropyl ring, or Rc and R d are taken together and are oxo(=O) or methylene(=CH2), or R c and R d are taken together with the atom to which they are attached to form spiro(C3-C5) cycloalkyl, R a , R b , R c and R d at least one of which is other than H, R 1 is H, hydroxy, cyano, halo, methoxycarbonyl, cyclopropyl, (C1-C3)alkyl, or (C1-C3)alkoxy, and any cyclopropyl, (C1-C3)alkyl, and (C1-C3)alkoxy may be substituted with one or more groups independently selected from hydroxy, (C1-C3)alkoxy, benzyloxy, cyano, and fluoro, alternatively R 1 and R c are taken together with the atom to which they are attached to form a fused cyclopropyl ring, each R 2 is independently selected from the group consisting of halo, cyano, hydroxy, ethynyl, (C1-C3)alkylsulfonyl, cyclopropyl, (C1-C3)alkyl, and (C1-C3)alkoxy, and any cyclopropyl, (C1-C3)alkyl, and (C1-C3)alkoxy may be substituted with one or more fluoro, R 4 is (C1-C7)alkyl, (C3-C7)cycloalkyl, phenyl, or 6-membered heteroaryl, and any R 4 may be substituted with one or more R x s, each R xis independently selected from the group consisting of hydroxy, halo, cyano, (C1-C5)alkyl, (C1-C5)alkoxy, (C3-C5)cyclopropyloxy, and (C3-C7)cycloalkyl, and any (C1-C5)alkyl, (C1-C5)alkoxy, (C3-C5)cyclopropyloxy, and (C3-C5)cycloalkyl may be substituted with one or more groups independently selected from halo, cyano, (C1-C5)alkyl, and hydroxy, and A is 0, 1, or 2.

[0009] In some embodiments, the disclosure provides a compound of formula I: TIFF2025522953000003.tif58165 or a salt thereof, wherein L 1 is CR a R b and L 2 is CR c R d and R a is H, halo, cyano, (C1-C5)alkyl, (C1-C5)alkoxy, (C3-C5)cyclopropyloxy, or (C3-C5)cycloalkyl, and any (C1-C5)alkyl, (C1-C5)alkoxy, (C3-C5)cyclopropyloxy, and (C3-C5)cycloalkyl may be substituted with one or more fluoros, R b is H, halo, cyano, (C1-C5)alkyl, (C1-C5)alkoxy, or (C3-C5)cycloalkyl, and any (C1-C5)alkyl, (C1-C5)alkoxy, and (C3-C5)cycloalkyl may be substituted with one or more fluoros, or R a and R b together are oxo(=O)methylene(=CH2), or R a and R b together with the atom to which they are attached form spiro(C3-C5)cycloalkyl, Rc is H, halo, cyano, (C1-C5)alkyl, (C1-C5)alkoxy, (C3-C5)cyclopropyloxy, or (C3-C5)cycloalkyl, and any (C1-C5)alkyl, (C1-C5)alkoxy, (C3-C5)cyclopropyloxy, and (C3-C5)cycloalkyl may be substituted with one or more fluoro, R d is H, halo, cyano, (C1-C5)alkyl, (C1-C5)alkoxy, or (C3-C5)cycloalkyl, and any (C1-C5)alkyl, (C1-C5)alkoxy, and (C3-C5)cycloalkyl may be substituted with one or more fluoro, or R c and R 1 together with the atom to which they are attached form a fused cyclopropyl ring, or R c and R d together are oxo(=O) or methylene(=CH2), or R c and R d together with the atom to which they are attached form a spiro(C3-C5)cycloalkyl, R 1 is (C1-C3)alkyl substituted with one or more fluoro, each R 2 is independently selected from the group consisting of halo, cyano, hydroxy, ethynyl, (C1-C3)alkylsulfonyl, cyclopropyl, (C1-C3)alkyl, and (C1-C3)alkoxy, and any cyclopropyl, (C1-C3)alkyl, and (C1-C3)alkoxy may be substituted with one or more fluoro, R 4 is (C1-C7)alkyl, (C3-C7)cycloalkyl, phenyl, or 6-membered heteroaryl, and any R 4 may be substituted with one or more R x s, each R xis independently selected from the group consisting of hydroxy, halo, cyano, (C1-C5)alkyl, (C1-C5)alkoxy, (C3-C5)cyclopropyloxy, and (C3-C7)cycloalkyl, and any (C1-C5)alkyl, (C1-C5)alkoxy, (C3-C5)cyclopropyloxy, and (C3-C5)cycloalkyl may be substituted with one or more groups independently selected from halo, cyano, and hydroxy, and A is 0, 1, or 2.

[0010] The present disclosure also provides a pharmaceutical composition comprising a compound of formula I or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

[0011] The present disclosure also provides a method for treating a condition associated with TRPV4 modulation in an animal, the method comprising administering to the animal a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0012] The present disclosure also provides a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in drug therapy.

[0013] The present disclosure also provides a compound of formula (I) or a pharmaceutically acceptable salt thereof for prophylactic or therapeutic treatment of a condition associated with the modulation of TRPV4.

[0014] The present disclosure also provides the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof for the preparation of a medicament for treating a condition associated with the modulation of TRPV4 in an animal.

[0015] The present disclosure also provides the processes and intermediates disclosed herein that are useful for preparing a compound of formula I or a salt thereof.

[0016] Certain compounds of formula (I) exhibit lower toxicity, improved efficacy, improved metabolic stability (e.g., against CYP3A4), lower levels of active circulating metabolites, and / or a higher safety margin.

Brief Description of the Drawings

[0017]

Figure 1

Modes for Carrying Out the Invention

[0018] Unless otherwise specified, the following definitions are used: Halo or halogen is fluoro, chloro, bromo, or iodo. Alkyl, alkoxy, etc. denote both straight-chain and branched groups, but reference to an individual radical such as propyl includes only the straight-chain radical, and branched-chain isomers such as isopropyl are specifically mentioned.

[0019] The term “alkyl,” by itself or as part of another substituent, unless otherwise indicated, means a straight-chain or branched-chain hydrocarbon radical having the number of carbon atoms indicated (i.e., C 1-5 means 1 to 5 carbons). Examples include (C1-C5)alkyl, (C2-C5)alkyl, (C1-C3)alkyl, (C2-C3)alkyl, and (C3-C5)alkyl. Examples of alkyl groups include methyl, ethyl, n-propyl, iso-propyl, n-butyl, t-butyl, iso-butyl, sec-butyl, and n-pentyl.

[0020] The term “alkoxy” refers to an alkyl group attached to the remainder of the molecule through an oxygen atom (“oxy”).

[0021] The term “carbocycle” refers to a fully saturated or partially unsaturated (non-aromatic) all-carbon ring having the specified number of carbons.

[0022] The term 6-membered heteroaryl refers to the following rings: Includes TIFF2025522953000004.tif32165.

[0023] As used herein, a wavy line that intersects a bond in a chemical structure TIFF2025522953000005.tif12165 indicates the point of attachment of the wavy bond that intersects a bond in the chemical structure to the remainder of the molecule.

[0024] The terms "treat", "treatment", or "treating" to the extent related to a disease or condition include inhibiting a disease or condition, eliminating a disease or condition, and / or alleviating one or more symptoms of a disease or condition. The terms "treat", "treatment", or "treating" also refer to both therapeutic and / or prophylactic treatment, or preventive measures, the purpose of which is to prevent or slow (mitigate) an undesired physiological change or disorder, such as the onset or spread of cancer. For example, beneficial or desired clinical outcomes include, but are not limited to, alleviation of symptoms, whether detectable or not, attenuation of the degree of a disease or disorder, stabilization (i.e., not worsening) of the state of a disease or disorder, delay or deceleration of disease progression, remission or temporary remission of a disease state or disorder, and remission (partial or complete). "Treat", "treatment", or "treating" can also mean extending the survival period as compared to the predicted survival period in the absence of treatment. Persons in need of treatment include those who already have a disease or disorder, as well as those who tend to have a disease or disorder, or those who need to prevent a disease or disorder. In some embodiments, "treat", "treatment", or "treating" does not include preventing or prophylaxis.

[0025] The phrase "therapeutically effective amount" or "effective amount" includes, but is not limited to, an amount of a compound of the present disclosure that (i) treats or prevents a particular disease, condition or disorder, (ii) attenuates, ameliorates or eliminates one or more symptoms of a particular disease, condition or disorder, or (iii) prevents or delays the onset of one or more symptoms of a particular disease, condition or disorder described herein.

[0026] In one aspect, the compounds of the present disclosure modulate TRPV4. In some embodiments, the modulation is antagonistic. In one aspect, the TRPV4 antagonists of the present disclosure are useful for treating conditions associated with overactivation of TRPV4. Such conditions include, for example, congenital distal spinal motor neuropathy, scapuloperoneal muscular atrophy, Charcot-Marie-Tooth disease type 2C (CMT2C), metatropic dysplasia (MD), spondyloepiphyseal dysplasia Kozlowski type (SMDK), spondyloepiphyseal dysplasia, Maroteaux type, pseudo-Morquio syndrome type 2, parastremmatic dysplasia, autosomal dominant short-trunk dwarfism, familial arthropathy of fingers with brachydactyly, and diseases caused by activating mutations of TRPV4, but are not limited thereto.

[0027] In one aspect, the TRPV4 antagonists of the present disclosure may be useful for treating related conditions associated with elevation of wild-type TRPV4 and / or symptoms overlapping with diseases caused by TRPV4 activating mutations. Such conditions include, but are not limited to, dysplasia, abnormal bone formation, osteoarthritis, decreased bone density, osteoporosis, peripheral neuropathy, hereditary neuropathy, spinal muscular atrophy, Charcot-Marie-Tooth disease, overactive bladder, urinary incontinence, interstitial cystitis, painful bladder syndrome, bladder urgency, and bladder diseases including neurogenic bladder.

[0028] In one aspect, the TRPV4 antagonists of the present disclosure are useful for treating other bone disorders including osteomyelitis, osteogenesis imperfecta, osteonecrosis, Paget's disease of bone, Creutzfeldt-Jakob disease, achondrogenesis, dwarfism, short stature, chondrodysplasia, and hereditary diseases of the skeleton. In one aspect, the TRPV4 antagonists may be useful for treating other neurological disorders including spinal muscular atrophy, neuropathic pain, pain, motor neuron disorders, chronic pain, abdominal pain, and spasm. In one aspect, the TRPV4 antagonists of the present disclosure are useful for treating disorders with impaired barrier integrity and dysregulated vascular permeability, including but not limited to retinal edema, retinal leakage, diabetes or other causes of macular edema, diabetic neuropathy, disorders associated with intestinal edema, postoperative abdominal edema, local and systemic edema, fluid retention, congestive heart failure, irritable bowel syndrome (IBS), Crohn's disease, diarrhea, intestinal abnormalities (hypersensitivity / hyporesponsiveness), fecal incontinence, constipation, celiac disease, lactose intolerance, and flatulence, obesity and type II diabetes (T2D), colitis / ulcerative colitis, hypertension, atherosclerosis, diseases with edema as a symptom, congestive heart failure, kidney disease or cirrhosis, kidney infection, urinary tract infection, and prostatic enlargement. In one aspect, the TRPV4 antagonists of the present disclosure are useful for treating lung diseases including but not limited to lung disorders, chronic obstructive pulmonary disorder, ventilator-induced lung injury, ventilator-associated lung parenchymal overinflation, high-altitude-induced pulmonary edema, and acute respiratory diseases. Useful for treating lung diseases including but not limited to constrictive syndrome, acute lung injury, pulmonary fibrosis, rhinosinusitis / rhinitis, asthma, cough, chronic cough, bronchiectasis, sarcoidosis, and pulmonary hypertension. In one aspect, the TRPV4 antagonists of the present disclosure are useful for treating inflammatory diseases including but not limited to sepsis, diseases with macrophage activation, microglial activation, neuroinflammation, Alzheimer's disease, Parkinson's disease, multiple sclerosis, and heart diseases, glaucoma, viral and bacterial infections, infectious diseases, chronic inflammatory diseases (rheumatoid arthritis), tissue repair, multiple organ dysfunction / multiple organ failure, microbial infections, acute brain / lung / liver / kidney injury, neurodegenerative diseases, tumor formation, cardiovascular diseases and metabolic diseases, and autoimmune diseases.

[0029] As used herein, the term "mammal" refers to humans, higher non-human primates, rodents, domestic animals, cows, horses, pigs, sheep, dogs, and cats. In some embodiments, the mammal is a human. As used herein, the term "patient" refers to any animal including a mammal. In one embodiment, the patient is a mammalian patient. In some embodiments, the patient is a human patient.

[0030] The compounds disclosed herein may exist in certain cases as tautomeric isomers. Only one delocalized resonance structure is illustrated, but all such forms are contemplated within the scope of this disclosure.

[0031] The present disclosure also includes, but is not limited to, any or all atoms enriched beyond the naturally occurring isotope ratios in one or more isotopes such as deuterium ( 2 H or D) in any claimed compound, as will be understood by those skilled in the art. By way of non-limiting example, a -CH3 group may be replaced by -CD3. In some embodiments, the compound is enriched in deuterium at a position one above the naturally occurring deuterium isotope ratio. In some embodiments, the compound is enriched in deuterium at a position two above the naturally occurring deuterium isotope ratio. In some embodiments, the compound is enriched in deuterium by at least about 75% at one position. In some embodiments, the compound is enriched in deuterium by at least about 75% at two positions. In some embodiments, the compound is enriched in deuterium by at least about 95% at one position. In some embodiments, the compound is enriched in deuterium by at least about 95% at two positions. The pharmaceutical compositions of the present disclosure include one or more excipients. When used in combination with the pharmaceutical compositions of the present disclosure, the term "excipient" generally refers to additional ingredients that, when combined with a compound of formula (I) or a pharmaceutically acceptable salt thereof, provide the corresponding composition. For example, when used in combination with the pharmaceutical compositions of the present disclosure, the term "excipient" includes, but is not limited to, carriers, binders, disintegrants, lubricants, sweeteners, flavoring agents, coating agents, preservatives, and coloring agents.

[0032] As used herein, the term "about" refers to an amount, value, or duration that is within ±10% of the recited amount, value, or duration. In some embodiments, "about" refers to an amount, value, or duration that is within ±10%, ±8%, ±6%, ±5%, ±4%, ±2%, ±1%, or ±0.5% of the recited amount, value, or duration. In some embodiments, "about" refers to an amount, value, or duration that is within ±10%, ±8%, ±6%, ±5%, ±4%, ±2% of the recited amount, value, or duration. In other embodiments, "about" refers to an amount, value, or duration that is within ±5% of the recited amount, value, or duration. In some embodiments, "about" refers to an amount, value, or duration that is within ±2% or ±1% of the recited amount, value, or duration. For example, in some embodiments, when the term "about" is used in connection with the recitation of a temperature or temperature range, these terms refer to ±5°C, ±2°C, or ±1°C of the recited temperature or temperature range. In other embodiments, the term "about" refers to ±2°C of the recited temperature or temperature range.

[0033] The stereochemical definitions and conventions used herein generally follow S.P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-Hill Book Company, New York and Eliel, E. and Wilen, S., “Stereochemistry of Organic Compounds”, John Wiley & Sons, Inc., New York, 1994. The compounds of the present disclosure may contain asymmetric or chiral centers and thus exist in different stereoisomers. All stereoisomers of the compounds of the present disclosure, including but not limited to diastereomers, enantiomers, and atropisomers, as well as mixtures thereof such as racemic mixtures, are intended to form part of the present invention.

[0034] The compounds of the present disclosure having a chiral center can exist as enantiomers and racemates, and it will be apparent to those skilled in the art that they can be isolated as enantiomers and racemates. Some compounds may exhibit polymorphism. It should be understood that the present disclosure encompasses any racemate, enantiomer, polymorph, or stereoisomer of the compounds of the present disclosure having the useful properties described herein, or mixtures thereof, and methods for preparing the optically active forms (e.g., by resolution of the racemate by recrystallization techniques, by synthesis from optically active starting materials, by chiral synthesis, or by chromatographic separation using a chiral stationary phase) are well known in the art.

[0035] When the bonds in the formula of the compounds herein are depicted in a non-stereochemical format (e.g., planar), the atoms to which the bonds are attached include all stereochemical possibilities. Boldface and dashed rectangles TIFF2025522953000006.tif17165 shows relative stereochemistry. When the bonds in the formula of the compounds herein are depicted as bold-wedge or dashed-wedge TIFF2025522953000007.tif17165, it should be understood that the atoms to which the stereochemical bonds are attached are enriched in the depicted absolute stereoisomer, unless otherwise noted. In some embodiments, the compound can be at least 51% of the depicted absolute stereoisomer. In some embodiments, the compound can be at least 60% of the depicted absolute stereoisomer. In some embodiments, the compound can be at least 80% of the depicted absolute stereoisomer. In some embodiments, the compound can be at least 90% of the depicted absolute stereoisomer. In some embodiments, the compound can be at least 95% of the depicted absolute stereoisomer. In some embodiments, the compound can be at least 99% of the depicted absolute stereoisomer.

[0036] Regarding radicals, substituents, and ranges, the specific values listed below are for illustrative purposes only and do not exclude other defined values or other values within the ranges defined for the radicals and substituents. It should be understood that two or more values can be combined. It should also be understood that the values (or subsets thereof) listed below can be excluded.

[0037] For the compounds of the present disclosure, the variable L 1 L 2 R a R b R c R d R 1 R 2 R 4 and R x each can be selected from the groups described herein, where applicable, and the variable L 1 L 2 R a R b R c R d R 1 R 2 R 4 and R x any group described herein for any of them can, where applicable, be combined with any group described herein for one or more of the remainder of the variable L 1 L 2 R a R b R c R d R 1 R 2 R 4 and R x It is understood that they can be combined.

[0038] Specifically, (C1-C5)alkyl can be methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, pentyl, or 3-pentyl, (C3-C6)cycloalkyl can be cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, and (C1-C5)alkoxy can be methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, pentoxy, or 3-pentoxy.

[0039] In some embodiments of the compounds of formula I, L 1 is CR a R b and L 2 is CR c R d and R a is H, halo, cyano, (C1-C5)alkyl, (C1-C5)alkoxy, (C3-C5)cycloalkyloxy, or (C3-C5)cycloalkyl, and any (C1-C5)alkyl, (C1-C5)alkoxy, (C3-C5)cycloalkyloxy, and (C3-C5)cycloalkyl may be substituted with one or more fluorines, R b is H, halo, cyano, (C1-C5)alkyl, (C1-C5)alkoxy, or (C3-C5)cycloalkyl, and any (C1-C5)alkyl, (C1-C5)alkoxy, and (C3-C5)cycloalkyl may be substituted with one or more fluorines, or R a and R b together are oxo(=O)methylene(=CH2), or R a and R b together with the atom to which they are attached form spiro(C3-C5)cycloalkyl, R cis H, halo, cyano, (C1-C5)alkyl, (C1-C5)alkoxy, (C3-C5)cyclopropyloxy, or (C3-C5)cycloalkyl, and any (C1-C5)alkyl, (C1-C5)alkoxy, (C3-C5)cyclopropyloxy, and (C3-C5)cycloalkyl may be substituted with one or more fluoro, R d is H, halo, cyano, (C1-C5)alkyl, (C1-C5)alkoxy, or (C3-C5)cycloalkyl, and any (C1-C5)alkyl, (C1-C5)alkoxy, and (C3-C5)cycloalkyl may be substituted with one or more fluoro, or R c and R 1 together with the atom to which they are attached form a fused cyclopropyl ring, or R c and R d together are oxo (=O) or methylene (=CH2), or R c and R d together with the atom to which they are attached form spiro (C3-C5)cycloalkyl, R a 、R b 、R c 、and R d at least one of which is other than H, R 1 is H, hydroxy, cyano, halo, methoxycarbonyl, cyclopropyl, (C1-C3)alkyl, or (C1-C3)alkoxy, and any cyclopropyl, (C1-C3)alkyl, and (C1-C3)alkoxy may be substituted with one or more groups independently selected from hydroxy, (C1-C3)alkoxy, benzyloxy, cyano, and fluoro, or R 1 and R c together with the atom to which they are attached form a fused cyclopropyl ring, each R 2is independently selected from the group consisting of halo, cyano, hydroxy, ethynyl, (C1-C3) alkylsulfonyl, cyclopropyl, (C1-C3) alkyl, and (C1-C3) alkoxy, and any cyclopropyl, (C1-C3) alkyl, and (C1-C3) alkoxy may be substituted with one or more fluorines, R 4 is (C1-C7) alkyl, (C3-C7) cycloalkyl, phenyl, or 6-membered heteroaryl, and any R 4 may be substituted with one or more R x groups, each R x is independently selected from the group consisting of hydroxy, halo, cyano, (C1-C5) alkyl, (C1-C5) alkoxy, and (C3-C7) cycloalkyl, and any (C1-C5) alkyl, (C1-C5) alkoxy, and (C3-C5) cycloalkyl may be substituted with one or more groups independently selected from halo, cyano, (C1-C5) alkyl, and hydroxy, and A is 0, 1, or 2.

[0040] In some embodiments of the compound of formula I, L 1 is CR a R b wherein, L 2 is CR c R d wherein, R a is H, halo, cyano, (C1-C5) alkyl, (C1-C5) alkoxy, (C3-C5) cycloalkyloxy, or (C3-C5) cycloalkyl, and any (C1-C5) alkyl, (C1-C5) alkoxy, (C3-C5) cycloalkyloxy, and (C3-C5) cycloalkyl may be substituted with one or more fluorines, R bis H, halo, cyano, (C1-C5)alkyl, (C1-C5)alkoxy, or (C3-C5)cycloalkyl, and any (C1-C5)alkyl, (C1-C5)alkoxy, and (C3-C5)cycloalkyl may be substituted with one or more fluorines, or R a and R b together form oxo(=O)methylene(=CH2), or R a and R b together with the atom to which they are attached form spiro(C3-C5)cycloalkyl, R c is H, halo, cyano, (C1-C5)alkyl, (C1-C5)alkoxy, (C3-C5)cycloalkyloxy, or (C3-C5)cycloalkyl, and any (C1-C5)alkyl, (C1-C5)alkoxy, (C3-C5)cycloalkyloxy, and (C3-C5)cycloalkyl may be substituted with one or more fluorines, R d is H, halo, cyano, (C1-C5)alkyl, (C1-C5)alkoxy, or (C3-C5)cycloalkyl, and any (C1-C5)alkyl, (C1-C5)alkoxy, and (C3-C5)cycloalkyl may be substituted with one or more fluorines, or R c and R 1 together with the atom to which they are attached form a fused cyclopropyl ring, or R c and R d together form oxo(=O) or methylene(=CH2), or R c and R d together with the atom to which they are attached form spiro(C3-C5)cycloalkyl, R 1 is (C1-C3)alkyl substituted with one or more fluorines, each R 2is independently selected from the group consisting of halo, cyano, hydroxy, ethynyl, (C1-C3) alkylsulfonyl, cyclopropyl, (C1-C3) alkyl, and (C1-C3) alkoxy, and any cyclopropyl, (C1-C3) alkyl, and (C1-C3) alkoxy may be substituted with one or more fluoros, R 4 is (C1-C7) alkyl, (C3-C7) cycloalkyl, phenyl, or 6-membered heteroaryl, and any R 4 may be substituted with one or more R x groups, each R x is independently selected from the group consisting of hydroxy, halo, cyano, (C1-C5) alkyl, (C1-C5) alkoxy, and (C3-C7) cycloalkyl, and any (C1-C5) alkyl, (C1-C5) alkoxy, and (C3-C5) cycloalkyl may be substituted with one or more groups independently selected from halo, cyano, and hydroxy, and A is 0, 1, or 2.

[0041] In some embodiments of the compounds of formula I, L 1 is CR a R b wherein, L 2 is CR c R d wherein, R a is H, halo, cyano, (C1-C5) alkyl, (C1-C5) alkoxy, or (C3-C5) cycloalkyl, and any (C1-C5) alkyl, (C1-C5) alkoxy, and (C3-C5) cycloalkyl may be substituted with one or more fluoros, R b is H, halo, cyano, (C1-C5) alkyl, (C1-C5) alkoxy, or (C3-C5) cycloalkyl, and any (C1-C5) alkyl, (C1-C5) alkoxy, and (C3-C5) cycloalkyl may be substituted with one or more fluoros, or R a and Rb together form oxo(=O)methylene(=CH2), or R a and R b together with the atom to which they are attached form spiro(C3-C5)cycloalkyl, R c is H, halo, cyano, (C1-C5)alkyl, (C1-C5)alkoxy, or (C3-C5)cycloalkyl, and any (C1-C5)alkyl, (C1-C5)alkoxy, and (C3-C5)cycloalkyl may be substituted with one or more fluoro, R d is H, halo, cyano, (C1-C5)alkyl, (C1-C5)alkoxy, or (C3-C5)cycloalkyl, and any (C1-C5)alkyl, (C1-C5)alkoxy, and (C3-C5)cycloalkyl may be substituted with one or more fluoro, or R c and R 1 together with the atom to which they are attached form a fused cyclopropyl ring, R c and R d together form oxo(=O) or methylene(=CH2), or R c and R d together with the atom to which they are attached form spiro(C3-C5)cycloalkyl, R a R b R c and R d at least one of is other than H, R 1 is H, hydroxy, cyano, halo, methoxycarbonyl, cyclopropyl, (C1-C3)alkyl, or (C1-C3)alkoxy, and any cyclopropyl, (C1-C3)alkyl, and (C1-C3)alkoxy may be substituted with one or more groups independently selected from hydroxy, (C1-C3)alkoxy, benzyloxy, cyano, and fluoro, or R 1 and R c together with the atom to which they are attached form a fused cyclopropyl ring, each R2 is independently selected from the group consisting of halo, cyano, hydroxy, ethynyl, (C1-C3) alkylsulfonyl, cyclopropyl, (C1-C3) alkyl, and (C1-C3) alkoxy, and any cyclopropyl, (C1-C3) alkyl, and (C1-C3) alkoxy may be substituted with one or more fluoros, R 4 is (C1-C7) alkyl, (C3-C7) cycloalkyl, phenyl, or 6-membered heteroaryl, and any R 4 may be substituted with one or more (e.g., 1, 2, 3, or 4) R x groups, each R x is independently selected from the group consisting of hydroxy, halo, cyano, (C1-C5) alkyl, (C1-C5) alkoxy, and (C3-C7) cycloalkyl, and any (C1-C5) alkyl, (C1-C5) alkoxy, and (C3-C5) cycloalkyl may be substituted with one or more groups independently selected from halo, cyano, (C1-C5) alkyl, and hydroxy, and A is 0, 1, or 2, or L 1 is CR a R b where L 2 is CR c R d where R a is H, halo, cyano, (C1-C5) alkyl, (C1-C5) alkoxy, or (C3-C5) cycloalkyl, and any (C1-C5) alkyl, (C1-C5) alkoxy, and (C3-C5) cycloalkyl may be substituted with one or more fluoros, R b is H, halo, cyano, (C1-C5) alkyl, (C1-C5) alkoxy, or (C3-C5) cycloalkyl, and any (C1-C5) alkyl, (C1-C5) alkoxy, and (C3-C5) cycloalkyl may be substituted with one or more fluoros, or R a and R btogether form oxo(=O)methylene(=CH2), or R a and R b together with the atom to which they are attached form spiro(C3-C5)cycloalkyl, R c is H, halo, cyano, (C1-C5)alkyl, (C1-C5)alkoxy, or (C3-C5)cycloalkyl, and any (C1-C5)alkyl, (C1-C5)alkoxy, and (C3-C5)cycloalkyl may be substituted with one or more fluoro, R d is H, halo, cyano, (C1-C5)alkyl, (C1-C5)alkoxy, or (C3-C5)cycloalkyl, and any (C1-C5)alkyl, (C1-C5)alkoxy, and (C3-C5)cycloalkyl may be substituted with one or more fluoro, or R c and R 1 together with the atom to which they are attached form a fused cyclopropyl ring, R c and R d together form oxo(=O) or methylene(=CH2), or R c and R d together with the atom to which they are attached form spiro(C3-C5)cycloalkyl, R 1 is (C1-C3)alkyl substituted with one or more fluoro, each R 2 is independently selected from the group consisting of halo, cyano, hydroxy, ethynyl, (C1-C3)alkylsulfonyl, cyclopropyl, (C1-C3)alkyl, and (C1-C3)alkoxy, and any cyclopropyl, (C1-C3)alkyl, and (C1-C3)alkoxy may be substituted with one or more fluoro, R 4 is (C1-C7)alkyl, (C3-C7)cycloalkyl, phenyl, or 6-membered heteroaryl, and any R 4 may be substituted with one or more (e.g., 1, 2, 3, or 4) R x s, Each R x is independently selected from the group consisting of hydroxy, halo, cyano, (C1-C5)alkyl, (C1-C5)alkoxy, and (C3-C7)cycloalkyl, and any (C1-C5)alkyl, (C1-C5)alkoxy, and (C3-C5)cycloalkyl may be substituted with one or more groups independently selected from halo, cyano, and hydroxy, and A is 0, 1, or 2.

[0042] In some embodiments of the compound of formula I, L 1 is CR a R b wherein L 2 is CR c R d wherein R a is H, halo, cyano, (C1-C5)alkyl, (C1-C5)alkoxy, or (C3-C5)cycloalkyl, and any (C1-C5)alkyl, (C1-C5)alkoxy, and (C3-C5)cycloalkyl may be substituted with one or more fluoro, R b is H, halo, cyano, (C1-C5)alkyl, (C1-C5)alkoxy, or (C3-C5)cycloalkyl, and any (C1-C5)alkyl, (C1-C5)alkoxy, and (C3-C5)cycloalkyl may be substituted with one or more fluoro, or R a and R b together are oxo(=O)methylene(=CH2), or R a and R b together with the atom to which they are attached form spiro(C3-C5)cycloalkyl spiro(C3-C5)cycloalkyl, R cis H, halo, cyano, (C1-C5)alkyl, (C1-C5)alkoxy, or (C3-C5)cycloalkyl, and any (C1-C5)alkyl, (C1-C5)alkoxy, and (C3-C5)cycloalkyl may be substituted with one or more fluorines, R d is H, halo, cyano, Rc(C1-C5)alkyl, (C1-C5)alkoxy, or (C3-C5)cycloalkyl, and any (C1-C5)alkyl, (C1-C5)alkoxy, and (C3-C5)cycloalkyl may be substituted with one or more fluorines, or R c and R 1 together with the atom to which they are attached form a fused cyclopropyl ring, R c and R d together are oxo(=O) or methylene(=CH2), or R c and R d together with the atom to which they are attached form spiro(C3-C5)cycloalkyl spiro(C3-C5)cycloalkyl, R a 、R b 、R c 、and R d at least one of which is other than H, R 1 is H, hydroxy, cyano, halo, methoxycarbonyl, cyclopropyl, (C1-C3)alkyl, or (C1-C3)alkoxy, and any cyclopropyl, (C1-C3)alkyl, and (C1-C3)alkoxy may be substituted with one or more groups independently selected from hydroxy, (C1-C3)alkoxy, benzyloxy, cyano, and fluorine, or R 1 and R c together with the atom to which they are attached form a fused cyclopropyl ring, each R 2is independently selected from the group consisting of halo, cyano, hydroxy, ethynyl, (C1-C3) alkylsulfonyl, cyclopropyl, (C1-C3) alkyl, and (C1-C3) alkoxy, and any cyclopropyl, (C1-C3) alkyl, and (C1-C3) alkoxy may be substituted with one or more fluorines, R 4 is (C1-C7) alkyl, (C3-C7) cycloalkyl, phenyl, or 6-membered heteroaryl, and any R 4 may be substituted with one or more (e.g., 1, 2, 3, or 4) R x groups, each R x is independently selected from the group consisting of hydroxy, halo, cyano, (C1-C5) alkyl, (C1-C5) alkoxy, and (C3-C7) cycloalkyl, and any (C1-C5) alkyl, (C1-C5) alkoxy, and (C3-C5) cycloalkyl may be substituted with one or more groups independently selected from halo, cyano, (C1-C5) alkyl, and hydroxy, and A is 0, 1, or 2.

[0043] In some embodiments of the compound of formula I, L 1 is CR a R b wherein, L 2 is CR c R d wherein, R a is H, halo, cyano, (C1-C5) alkyl, (C1-C5) alkoxy, or (C3-C5) cycloalkyl, and any (C1-C5) alkyl, (C1-C5) alkoxy, and (C3-C5) cycloalkyl may be substituted with one or more fluorines, R bis H, halo, cyano, (C1-C5)alkyl, (C1-C5)alkoxy, or (C3-C5)cycloalkyl, and any (C1-C5)alkyl, (C1-C5)alkoxy, and (C3-C5)cycloalkyl may be substituted with one or more fluoros, or R a and R b together form oxo(=O)methylene(=CH2), or R a and R b together with the atom to which they are attached form spiro(C3-C5)cycloalkyl spiro(C3-C5)cycloalkyl, R c is H, halo, cyano, (C1-C5)alkyl, (C1-C5)alkoxy, or (C3-C5)cycloalkyl, and any (C1-C5)alkyl, (C1-C5)alkoxy, and (C3-C5)cycloalkyl may be substituted with one or more fluoros, R d is H, halo, cyano, Rc(C1-C5)alkyl, (C1-C5)alkoxy, or (C3-C5)cycloalkyl, and any (C1-C5)alkyl, (C1-C5)alkoxy, and (C3-C5)cycloalkyl may be substituted with one or more fluoros, or R c and R 1 together with the atom to which they are attached form a fused cyclopropyl ring, R c and R d together form oxo(=O)methylene(=CH2), or R c and R d together with the atom to which they are attached form spiro(C3-C5)cycloalkyl spiro(C3-C5)cycloalkyl, R 1 is (C1-C3)alkyl substituted with one or more fluoros, each R 2is independently selected from the group consisting of halo, cyano, hydroxy, ethynyl, (C1-C3) alkylsulfonyl, cyclopropyl, (C1-C3) alkyl, and (C1-C3) alkoxy, and any cyclopropyl, (C1-C3) alkyl, and (C1-C3) alkoxy may be substituted with one or more fluoros, R 4 is (C1-C7) alkyl, (C3-C7) cycloalkyl, phenyl, or 6-membered heteroaryl, and any R 4 may be substituted with one or more (e.g., 1, 2, 3, or 4) R x groups, each R x is independently selected from the group consisting of hydroxy, halo, cyano, (C1-C5) alkyl, (C1-C5) alkoxy, and (C3-C7) cycloalkyl, and any (C1-C5) alkyl, (C1-C5) alkoxy, and (C3-C5) cycloalkyl may be substituted with one or more groups independently selected from halo, cyano, and hydroxy, and A is 0, 1, or 2.

[0044] In some embodiments, the compound of formula (I) or a salt thereof is a compound of formula (II): TIFF2025522953000008.tif63165 a compound or a salt thereof, wherein B is 0, 1, or 2.

[0045] In some embodiments of the compound of formula (II), R 4 is (C3-C7) cycloalkyl, phenyl, or 6-membered heteroaryl.

[0046] In some embodiments, the compound of formula (I) or a salt thereof is a compound of formula (III): TIFF2025522953000009.tif63165 a compound or a salt thereof, wherein B is 0, 1, or 2.

[0047] In some embodiments of the compound of formula (III), R 4is (C3-C7) cycloalkyl, phenyl, or 6-membered heteroaryl.

[0048] In some embodiments, R a is H.

[0049] In some embodiments, R a is selected from the group consisting of halo, cyano, (C1-C5) alkyl, (C1-C5) alkoxy, and (C3-C5) cycloalkyl, and any (C1-C5) alkyl, (C1-C5) alkoxy, and (C3-C5) cycloalkyl may be substituted with one or more fluoros.

[0050] In some embodiments, R a is H, halo, cyano, CF3, CF2H, CFH2, OCF3, OCF2H, OCH2CF3, OCH2CF2H, (C1-C5) alkyl, or (C3-C5) cycloalkyl.

[0051] In some embodiments, R a is halo, cyano, CF3, CF2H, CFH2, OCF3, OCF2H, OCH2CF3, OCH2CF2H, (C1-C5) alkyl, or (C3-C5) cycloalkyl.

[0052] In some embodiments, R b is H.

[0053] In some embodiments, R b is halo, cyano, (C1-C5) alkyl, (C1-C5) alkoxy, and (C3-C5) cycloalkyl, and any (C1-C5) alkyl, (C1-C5) alkoxy, and (C3-C5) cyclopropyl may be substituted with one or more fluoros.

[0054] In some embodiments, R bis H, halo, cyano, CF3, CF2H, CFH2, OCF3, OCF2H, OCH2CF3, OCH2CF2H, (C1-C5)alkyl, or (C3-C5)cycloalkyl.

[0055] In some embodiments, R b is halo, cyano, CF3, CF2H, CFH2, OCF3, OCF2H, OCH2CF3, OCH2CF2H, (C1-C5)alkyl, or (C3-C5)cycloalkyl.

[0056] In some embodiments, R c is H.

[0057] In some embodiments, R c is selected from the group consisting of halo, cyano, (C1-C5)alkyl, (C1-C5)alkoxy, and (C3-C5)cycloalkyl, and any (C1-C5)alkyl, (C1-C5)alkoxy, and (C3-C5)cycloalkyl may be substituted with one or more fluoro.

[0058] In some embodiments, R c is H, halo, cyano, CF3, CF2H, CFH2, OCF3, OCF2H, OCH2CF3, OCH2CF2H, (C1-C5)alkyl, or (C3-C5)cycloalkyl.

[0059] In some embodiments, R c is halo, cyano, CF3, CF2H, CFH2, OCF3, OCF2H, OCH2CF3, OCH2CF2H, (C1-C5)alkyl, or (C3-C5)cycloalkyl.

[0060] In some embodiments, R 1 and R c together with the atom to which they are attached form a fused cyclopropyl ring.

[0061] In some embodiments, R d is H.

[0062] In some embodiments, R d is selected from the group consisting of halo, cyano, (C1-C5)alkyl, (C1-C5)alkoxy, and (C3-C5)cycloalkyl, and any (C1-C5)alkyl, (C1-C5)alkoxy, and (C3-C5)cycloalkyl may be substituted with one or more fluoro.

[0063] In some embodiments, R d is H, halo, cyano, CF3, CF2H, CFH2, OCF3, OCF2H, OCH2CF3, OCH2CF2H, (C1-C5)alkyl, or (C3-C5)cycloalkyl.

[0064] In some embodiments, R d is halo, cyano, CF3, CF2H, CFH2, OCF3, OCF2H, OCH2CF3, OCH2CF2H, (C1-C5)alkyl, or (C3-C5)cycloalkyl.

[0065] In some embodiments, R c and R d together with the atom to which they are attached form spiro(C3-C5)cycloalkyl.

[0066] In some embodiments, R 1 is selected from the group consisting of cyano, halo, methoxycarbonyl, cyclopropyl, (C1-C3)alkyl, and (C1-C3)alkoxy, and any cyclopropyl, (C1-C3)alkyl, and (C1-C3)alkoxy may be substituted with one or more groups independently selected from hydroxy, (C1C3)alkoxy, benzyloxy, and fluoro.

[0067] In some embodiments, R 1 is H, methyl, cyclopropyl, CF3, CF2H, CFH2, OCF3, or OCF2H.

[0068] In some embodiments, R 1 is hydroxy, hydroxymethyl, cyanomethyl.

[0069] In some embodiments, R 1 is methyl.

[0070] In some embodiments, R 1 is H.

[0071] In some embodiments, the group: TIFF2025522953000010.tif27165 is selected from the group consisting of TIFF2025522953000011.tif32165.

[0072] In some embodiments, A is 0.

[0073] In some embodiments, A is 1.

[0074] In some embodiments, R 2 is cyano.

[0075] In some embodiments, R 2 is 6-cyano.

[0076] In some embodiments, R 4 is phenyl which may be substituted with one or more R x s.

[0077] In some embodiments, R 4 is selected from the group consisting of TIFF2025522953000012.tif48165.

[0078] In some embodiments, R 4 is pyrimidin-2-yl which may be substituted with one or more fluoros.

[0079] In some embodiments, R 4 is selected from the group consisting of TIFF2025522953000013.tif27165.

[0080] In some embodiments, R 4 is (C1-C7) alkyl which may be substituted with one R x .

[0081] In some embodiments, R 4 is (C3-C7) cycloalkyl which may be substituted with one R x .

[0082] In some embodiments, R 4 is 6-membered heteroaryl which may be substituted with one R x .

[0083] In some embodiments, R x is C1-C5 alkyl which may be substituted with one or more groups independently selected from halo, cyano, and hydroxy.

[0084] In some embodiments, R x is C1-C5 alkyl substituted with hydroxy.

[0085] In some embodiments, R x is 2-hydroxy-2-methylethyl.

[0086] In some embodiments, the compound of formula (I) or a salt thereof is a compound of formula IV or V: TIFF2025522953000014.tif73165 or a salt thereof, wherein B is 0, 1, or 2.

[0087] In some embodiments, the compound of formula (I) or a salt thereof is a compound of formula IV or Va: The compound of TIFF2025522953000015.tif73165 or a salt thereof, wherein B is 0, 1, or 2.

[0088] In some embodiments, the compound of formula (I) or a salt thereof is of formula VI or VII: The compound of TIFF2025522953000016.tif68165 or a salt thereof.

[0089] In some embodiments, R c is fluoro and R d is H or fluoro.

[0090] In some embodiments, the compound or a salt thereof is the following: Selected from the group consisting of TIFF2025522953000017.tif207165 TIFF2025522953000018.tif70165 and salts thereof.

[0091] In some embodiments, the compound or a salt thereof is the following: Selected from the group consisting of TIFF2025522953000019.tif74165 and salts thereof.

[0092] In some embodiments, the compound or a salt thereof is the following: Selected from the group consisting of TIFF2025522953000020.tif68165 and salts thereof.

[0093] In some embodiments, the compound or a salt thereof is the following: Selected from the group consisting of TIFF2025522953000021.tif63165 and salts thereof.

[0094] In some embodiments, the compound or a salt thereof is the following: Selected from the group consisting of TIFF2025522953000022.tif125165 and salts thereof.

[0095] In some embodiments, the compound or a salt thereof is: TIFF2025522953000023.tif63165 or a salt thereof.

[0096] In some embodiments, the compound or a salt thereof is: TIFF2025522953000024.tif63165 or a salt thereof.

[0097] In some embodiments, the present disclosure provides 1-(((5S,7S,8R)-8-fluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile, or a pharmaceutically acceptable salt thereof.

[0098] In some embodiments, the present disclosure provides a compound of formula (I) or a salt thereof having lower toxicity than GSK2798745.

[0099] In some embodiments, the present disclosure provides a compound of formula (I) or a salt thereof having improved potency compared to GSK2798745.

[0100] In some embodiments, the present disclosure provides a compound of formula (I) or a salt thereof having improved metabolic stability (e.g., against CYP3A4) compared to GSK2798745.

[0101] In some embodiments, the present disclosure provides a compound of formula (I) or a salt thereof having a lower level of active circulating metabolites than GSK2798745.

[0102] In some embodiments, the present disclosure provides a compound of formula (I) or a salt thereof having a higher safety ceiling than GSK2798745.

[0103] In some embodiments, the present disclosure provides a compound of formula (I) or a salt thereof having improved metabolic stability (e.g., against CYP3A4) compared to GSK2798745 and substantially the same or improved efficacy.

[0104] In some embodiments, the present disclosure provides a compound of formula (I) or a salt thereof having improved metabolic stability (e.g., against CYP3A4) compared to GSK2798745 and substantially the same or improved efficacy.

[0105] In some embodiments, the present disclosure provides a solid of 1-(((5S,7S,8R)-8-fluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile having one or more XRPD diffraction signals at about 7.04, about 7.07, about 7.09, about 10.33, about 10.35, about 18.87, and about 24.02° 2θ. In some embodiments, the present disclosure provides a solid of 1-(((5S,7S,8R)-8-fluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile having an XRPD diffraction signal at about 7.04, about 7.07, about 7.09, about 10.33, about 10.35, about 18.87, and about 24.02 ° 2θ. With respect to the XRPD peaks, the term "about" means ±0.01° 2θ.

[0106] In some embodiments, the present disclosure provides 1-(((5S,7S,8R)-8-fluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile having an XRPD diffraction pattern substantially the same as that shown in FIG. 1. In some embodiments, the term "substantially the same" refers to an XRPD diffraction pattern having at least three similar signals.

[0107] A process for preparing the compound of formula I is provided as a further embodiment of the present disclosure and is described by the following procedure. Here, the meaning of the general radicals is as given above unless otherwise modified.

[0108] If the compound is sufficiently basic or acidic, the salts of the compound of formula I may be useful as intermediates for isolating or purifying the compound of formula I. Also, administration of the compound of formula I as a pharmaceutically acceptable acid or basic salt may be appropriate. Examples of pharmaceutically acceptable salts include organic acid addition salts formed with acids that form physiologically acceptable anions, such as tosylate, methanesulfonate, acetate, citrate, malonate, tartrate, succinate, benzoate, ascorbate, α-ketoglutaric acid, and α-glycerophosphoric acid. Suitable inorganic salts, including hydrochloride, sulfate, nitrate, bicarbonate, and carbonate, may also be formed.

[0109] Salts can be obtained using standard procedures well known in the art, for example, by reacting a sufficiently basic compound, such as an amine, with a suitable acid that provides a physiologically acceptable anion. Alkali metal (e.g., sodium, potassium, or lithium) salts or alkaline earth metal (e.g., calcium) salts of carboxylic acids can also be produced.

[0110] The compounds of formula I are formulated as pharmaceutical compositions and administered to mammalian hosts such as human patients in various forms adapted to the selected route of administration, i.e., orally or parenterally, by intravenous, intramuscular, topical or subcutaneous routes.

[0111] Accordingly, the compounds may be administered systemically, for example, orally, in combination with a pharmaceutically acceptable vehicle such as an inert diluent or assimilable edible carrier. They may be enclosed in hard or soft shell gelatin capsules, compressed into tablets, or incorporated directly into the food of the patient's diet. For oral therapeutic administration, the active compound may be incorporated with one or more excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, etc. Such compositions and preparations should contain at least 0.1% of the active compound. The percentage of the compositions and preparations can, of course, be varied and conveniently may be about 2 to 60% by weight of the given unit dosage form. The amount of the active compound in such therapeutically useful compositions is such that an effective dosage level is obtained.

[0112] Tablets, troches, pills, capsules, etc. may also contain the following: binders such as gum tragacanth, acacia, corn starch or gelatin; excipients such as dicalcium phosphate, disintegrants such as corn starch, potato starch, alginic acid, lubricants such as magnesium stearate, and sweetening agents such as sucrose, fructose, lactose, or aspartame, or flavoring agents such as peppermint, wintergreen oil, or cherry flavor may be added. If the unit dosage form is a capsule, it may contain a liquid carrier such as vegetable oil or polyethylene glycol in addition to the materials of the above type. Various other materials may be present as coatings or in other ways to modify the physical form of the solid unit dosage form. For example, tablets, pills, or capsules can be coated with gelatin, wax, shellac, or sugar. Syrups or elixirs may contain the active compound as a sweetening agent, sucrose or fructose, methyl and propyl parabens as preservatives, dyes and flavorings, e.g., cherry or orange flavor. Of course, any material used in preparing any unit dosage form must be pharmaceutically acceptable and substantially non-toxic in the amounts used. Further, the active compounds can be incorporated into sustained release formulations and devices.

[0113] The active compounds may also be administered intravenously or intraperitoneally by infusion or injection. Solutions of the active compounds or their salts can be prepared in water optionally mixed with a non-toxic surfactant. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, triacetin, and mixtures thereof and / or in oils. These preparations contain a preservative to prevent the growth of microorganisms under normal storage and use conditions.

[0114] Pharmaceutical dosage forms suitable for injection or infusion include sterile injectable or insoluble solutions, or sterile aqueous solutions or dispersions or sterile powders containing an active ingredient suitable for the immediate preparation of a dispersion optionally encapsulated in liposomes. In all cases, the final dosage form must be sterile and stable under the manufacturing and storage conditions. The liquid carrier can be, for example, a solvent or liquid dispersion medium containing water, ethanol, polyols (such as glycerol, propylene glycol, liquid polyethylene glycol, etc.), vegetable oils, non-toxic glyceryl esters, and suitable mixtures thereof. The inhibition of the action of microorganisms can be brought about by various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, etc. In many cases, it is preferable to include an isotonic agent, such as sugar, buffer, or sodium chloride. The prolongation of the absorption of injectable compositions can be brought about by the use in the composition of agents that delay absorption, such as aluminum monostearate and gelatin.

[0115] Sterile injectable solutions are prepared by incorporating the active compound in the required amount into a suitable solvent together with the various other ingredients described above, and, if necessary, subsequently filtering the solution aseptically. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred method of preparation is by vacuum drying and lyophilization techniques, which yield a powder that combines the active ingredient present in a previously aseptically filtered solution with any desired additional ingredients.

[0116] For topical administration, the compounds of the present invention can be applied in pure form, i.e., when they are liquid. However, in general, it is desirable to administer them topically as a composition or formulation in combination with a dermatologically acceptable carrier, which can be solid or liquid.

[0117] Useful solid carriers include fine solids such as talc, clay, crystalline cellulose, silica, alumina, etc. Useful liquid carriers include water, alcohol, or glycol, or water-alcohol / glycol mixtures, in which the compound can be dissolved or dispersed at an effective level, optionally with the aid of a non-toxic surfactant. Auxiliaries such as fragrances and additional antibacterial agents can be added to optimize the properties for a given use. The resulting liquid composition can be applied from an absorbent pad and used to impregnate bandages and other dressings, or can be sprayed onto the affected area using a pump-type or aerosol sprayer.

[0118] Thickeners such as synthetic polymers, fatty acids, fatty acid salts and esters, aliphatic alcohols, modified celluloses, or modified mineral materials can also be used with the liquid carrier to form a paintable paste, gel, ointment, soap, etc. for direct application to the skin of the user.

[0119] Examples of useful skin compositions for delivering the compound of formula I are known in the art; see, for example, Jacquet et al. (U.S. Patent No. 4,608,392), Geria (U.S. Patent No. 4,992,478), Smith et al. (U.S. Patent No. 4,559,157), and Wortzman (U.S. Patent No. 4,820,508).

[0120] Useful dosages of the compound of formula I can be determined by comparing their in vitro and in vivo activities in animal models. Methods for extrapolating effective dosages in mice and other animals to humans are known to those skilled in the art; see, for example, U.S. Patent No. 4,938,949.

[0121] The amount of the compound, or its active salt or derivative, required for use in therapy will vary not only according to the particular salt selected but also according to the route of administration, the nature of the condition being treated and the age and condition of the subject, and will be determined in accordance with the condition of the patient and ultimately at the discretion of the attending physician or clinician.

[0122] The desired dosage may conveniently be administered as a single dose, or as divided doses at appropriate intervals, for example, two, three, four or more divided doses per day. The divided doses themselves may also be further divided, for example, into several discrete loose spaced administrations. Examples include multiple inhalations from an inhaler, or multiple applications for administration to the eye.

[0123] The compounds of the present disclosure may be administered alone or in combination with one or more other therapeutic agents, for example, endothelin receptor antagonists, angiotensin converting enzyme (ACE) inhibitors, angiotensin II receptor antagonists, vasopeptidase inhibitors, vasopressin receptor modulators, diuretics, digoxin, beta blockers, aldosterone antagonists, inotropes, NSAIDS, nitric oxide donors, calcium channel modulators, muscarinic antagonists, anti-inflammatory steroid agents, bronchodilators, antihistamines, leukotriene antagonists, HMG-CoA reductase inhibitors, non-selective adrenergic and α1-adrenergic receptor dual antagonists, type 5 phosphodiesterase inhibitors, and renin inhibitors, selected from the group consisting of one or more agents.

[0124] Accordingly, in some embodiments, the present disclosure also provides a composition comprising a compound of formula I, or a pharmaceutically acceptable salt thereof, at least one other therapeutic agent, and a pharmaceutically acceptable diluent or carrier. The present disclosure also provides a kit for treating a condition associated with the modulation of TRPV4, comprising a compound of formula I or a pharmaceutically acceptable salt thereof, at least one other therapeutic agent, a packaging material, and instructions for administering the compound of formula I or a pharmaceutically acceptable salt thereof and other therapeutic agent(s) to an animal.

[0125] The biological activity of the compound can be determined using any suitable assay for evaluating TRPV4 antagonist activity, as well as any suitable tissue or in vivo model. For example, the biological activity of the compound can be evaluated using the assay described in Example 23 herein, or using the assays and models described in International Patent Application Publication Nos. WO2012 / 174340, WO2012 / 174342, WO2013 / 012500, and WO2017 / 199199.

[0126] Method of Use In some embodiments, the present disclosure provides a method of modulating TRPV4 expression using a compound of the present disclosure or a salt thereof.

[0127] In some embodiments, the present disclosure provides a method of modulating the expression of TRPV4 with a pharmaceutical composition comprising a compound of the present disclosure or a salt thereof.

[0128] In some embodiments, the present disclosure provides a method of treating or preventing a disease or disorder disclosed herein in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of the present disclosure or a salt thereof.

[0129] In some embodiments, a method of treating or preventing a disease or disorder disclosed herein in a subject in need thereof, the method comprising administering a therapeutically effective amount of a compound or a salt thereof of the present disclosure, or a pharmaceutical composition of the present disclosure.

[0130] In some embodiments, the present disclosure provides a method of treating or preventing a disease or disorder disclosed herein in a subject in need thereof, the method comprising administering to the subject a compound of the present disclosure or a salt thereof.

[0131] In some embodiments, the present disclosure provides a method of treating or preventing a disease or disorder disclosed herein in a subject in need thereof, the method comprising administering a compound of the present disclosure or a salt thereof, or a pharmaceutical composition of the present disclosure.

[0132] In some embodiments, the present disclosure provides a compound of the present disclosure or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition, for use in modulating TRPV4 expression.

[0133] In some embodiments, the present disclosure provides a compound of the present disclosure or a salt thereof, or a pharmaceutical composition of the present disclosure, for use in the treatment or prevention of a disease or disorder.

[0134] In some embodiments, the present disclosure provides a compound of the present disclosure or a pharmaceutically acceptable salt thereof for use in the treatment or prevention of a disease or disorder disclosed herein.

[0135] In some embodiments, the present disclosure provides a compound of the present disclosure or a salt thereof for use in the treatment of a disease or disorder disclosed herein.

[0136] In some embodiments, the present disclosure provides the use of a compound of the present disclosure or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for modulating TRPV4 expression.

[0137] In some embodiments, the present disclosure provides the use of a compound of the present disclosure or a salt thereof in the manufacture of a medicament for treating or preventing a disease or disorder of the present disclosure.

[0138] In some embodiments, the present disclosure provides the use of a compound of the present disclosure or a salt thereof in the manufacture of a medicament for treating a disease or disorder of the present disclosure.

[0139] In some embodiments, the disease or disorder is related to TRPV4 involved.

[0140] In some embodiments, the disclosure provides a method for treating a condition associated with TRPV4 modulation, comprising administering a compound of the disclosure, a pharmaceutically acceptable salt thereof, or a solid thereof.

[0141] In some embodiments, the disclosure provides a compound of the disclosure, a salt thereof, or a solid thereof for use in a pharmaceutical therapy.

[0142] In some embodiments, the disclosure provides a compound of the disclosure, a salt or solid thereof for prophylactic or therapeutic treatment of a condition associated with TRPV4 modulation.

[0143] In some embodiments, the disclosure provides the use of a compound of the disclosure, a salt or solid thereof, for preparing a medicament for treating a condition associated with TRPV4 modulation in an animal.

[0144] In some embodiments, the salt is a pharmaceutically acceptable salt thereof.

[0145] In some embodiments, the compound is administered to a subject.

[0146] In some embodiments, the compound is administered to an animal.

[0147] In some embodiments, the subject is an animal.

[0148] In some embodiments, the subject is a human.

[0149] In some embodiments, the disease or disorder is a respiratory disease or disorder.

[0150] In some embodiments, the disease or disorder is chronic cough, pulmonary edema, chronic obstructive pulmonary disease or pulmonary fibrosis.

[0151] In some embodiments, the disease or disorder is inflammatory pain hypersensitivity.

[0152] In some embodiments, the disease or disorder is mechanical pain.

[0153] In some embodiments, the disease is neuropathic pain.

[0154] In some embodiments, the disease or disorder is a bladder-related condition.

[0155] In some embodiments, the disease or disorder is a respiratory disease, a joint disease, pain, or a bladder dysfunction.

[0156] In some embodiments, the disease or disorder is a joint disease.

[0157] In some embodiments, the disease or disorder is pain.

[0158] In some embodiments, the disease or disorder is a bladder dysfunction.

[0159] In some embodiments, the disease or disorder is a Mendelian disease.

[0160] In some embodiments, the disease or disorder is a skeletal formation abnormality.

[0161] In some embodiments, the regulation is inhibition.

[0162] In some embodiments, the regulation is antagonism.

[0163] Method of synthesis The compounds of formula (I) can be prepared using the synthetic methods and intermediates described in the following examples. The compounds of formula (I) can also be prepared using well-known synthetic methods and intermediate compounds, including, for example, the synthetic methods and intermediate compounds described in International Patent Application Publication Nos. WO2012 / 174340, WO2012 / 174342, WO2013 / 012500, and WO2017 / 199199. Schemes 1-6 illustrate the intermediates and methods that can be used to prepare the compounds of formula (I).

[0164] Scheme 1 TIFF2025522953000025.tif207165TIFF2025522953000026.tif73165

[0165] Scheme 2 TIFF2025522953000027.tif197165

[0166] Scheme 3 TIFF2025522953000028.tif68165

[0167] Scheme 4 TIFF2025522953000029.tif53165

[0168] Scheme 5 TIFF2025522953000030.tif125165

[0169] Scheme 6 TIFF2025522953000031.tif99165

[0170] Scheme 7 TIFF2025522953000032.tif130165

[0171] Scheme 8 TIFF2025522953000033.tif58165

[0172] Scheme 9 TIFF2025522953000034.tif42165

[0173] The present disclosure is further illustrated by the following non-limiting examples.

Example

[0174] For illustrative purposes, the neutral compound of formula (I) is synthesized and tested in the examples. The neutral compound of formula (I) can be converted to the salt of the corresponding compound using established techniques in the art (e.g., by saponification of an ester to the carboxylate, or by hydrolyzing an amide to form the corresponding carboxylic acid and then converting the carboxylic acid to the carboxylate).

[0175] Example 1. Preparation: 1-(((5S,7R)-3-(5-(2-Hydroxypropan-2-yl)pyrazin-2-yl)-8,8-dimethyl-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000035.tif63165

[0176] Reaction Scheme TIFF2025522953000036.tif202165TIFF2025522953000037.tif135165

[0177] Detailed Procedure rac-4,4-Dimethyl-3-(nitromethyl)cyclohexan-1-one TIFF2025522953000038.tif37165 Nitromethane (3.83 g, 62.81 mmol, 1.2 equiv) was added to a solution of 4,4-dimethylcyclohex-2-en-1-one (6.50 g, 52.34 mmol, 1 equiv) and benzyltrimethylammonium hydroxide (13.13 g, 78.51 mmol, 1.5 equiv) in methanol (60 mL). The resulting mixture was stirred at room temperature overnight and concentrated under vacuum to give the crude product. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0% - 10% petroleum ether. The fractions were combined and concentrated under vacuum to give the desired product 4,4-dimethyl-3-(nitromethyl)cyclohexan-1-one (6.70 g, 46% yield) as a pale yellow oil.

[0178] rac-8,8-dimethyl-7-(nitromethyl)-1,4-dioxaspiro[4.5]decane TIFF2025522953000039.tif42165 Ethylene glycol (3.37 g, 54.26 mmol, 1.5 equiv) was added to a solution of 4,4-dimethyl-3-(nitromethyl)cyclohexan-1-one (6.70 g, 36.17 mmol, 1 equiv) and trimethoxymethane (5.76 g, 54.26 mmol, 1.5 equiv) in dichloromethane (100 mL). The resulting reaction mixture was stirred at room temperature for 5 minutes and then cooled to 0 °C in an ice bath. Methanesulfonic acid (0.52 g, 5.42 mmol, 0.15 equiv) was added dropwise to this mixture. The resulting mixture was removed from the bath, warmed to room temperature, and stirred for 2 hours. The reaction mixture was concentrated under vacuum. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0% - 10% petroleum ether. The fractions were combined and concentrated under vacuum to give the desired product rac-8,8-dimethyl-7-(nitromethyl)-1,4-dioxaspiro[4.5]decane (6 g, 72.3% yield) as a pale yellow oil. 11H NMR (400 MHz, DMSO-d6) δ 4.73 (dd, J = 12.6, 3.9 Hz, 1H), 4.25 (dd, J = 12.6, 10.1 Hz, 1H), 3.85 - 3.82 (m, 4H), 2.17 (dd, J = 12.0, 10.1 Hz, 1H), 1.58 - 1.42 (m, 5H), 1.39 - 1.33 (m, 1H), 0.96 (s, 3H), 0.80 (s, 3H).

[0179] rac-(8,8-dimethyl-1,4-dioxaspiro[4.5]decan-7-yl)methanamine Palladium on carbon (10%, 0.28 g, 2.62 mmol, 0.1 equiv) was added to a solution of rac-8,8-dimethyl-7-(nitromethyl)-1,4-dioxaspiro[4.5]decane (6.0 g, 26.17 mmol, 1 equiv) in methanol (60 mL). The resulting mixture was stirred overnight at room temperature under a hydrogen atmosphere. The resulting mixture was filtered and the filter cake was washed with methanol (3 × 100 mL). The filtrate was concentrated under reduced pressure to afford the crude product rac-(8,8-dimethyl-1,4-dioxaspiro[4.5]decan-7-yl)methanamine (crude 6 g) as a colorless oil. LCMS (ESI-MS) m / z = 200 [M+H] + .

[0180] rac-3-(((8,8-dimethyl-1,4-dioxaspiro[4.5]decan-7-yl)methyl)amino)-4-nitrobenzonitrile TIFF2025522953000041.tif53165 Potassium carbonate (8.32 g, 60.21 mmol, 2 equiv) was added to a solution of rac-(8,8-dimethyl-1,4-dioxaspiro[4.5]decan-7-yl)methanamine (6 g, 30.1 mmol, 1 equiv) and 3-fluoro-4-nitrobenzonitrile (5 g, 30.1 mmol, 1 equiv) in acetonitrile (60 mL). The resulting mixture was stirred at room temperature overnight. The reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (3 x 100 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to give the crude product. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0% - 10% petroleum ether, and concentrated under vacuum to give the desired product rac-3-(((8,8-dimethyl-1,4-dioxaspiro)[4.5]decan-7-yl)methyl)amino)-4-nitrobenzonitrile (7 g, 67.3% yield) as an orange solid. 1 1H NMR (400 MHz, DMSO-d6) δ 8.38 - 8.31 (m, 1H), 8.18 (dd, J = 17.2, 8.6 Hz, 1H), 8.10 - 7.96 (m, 2H), 3.94 - 3.79 (m, 4H), 3.55 - 3.49 (m, 1H), 3.14 - 3.06 (m, 1H), 1.85 - 1.30 (m, 7H), 1.07 - 0.89 (m, 6H). LCMS (ESI-MS) m / z = 346 [M+H] + .

[0181] rac-4-amino-3-(((8,8-dimethyl-1,4-dioxaspiro[4.5]decan-7-yl)methyl)amino)benzonitrile TIFF2025522953000042.tif Palladium on carbon (10%, 0.22 g, 2.03 mmol, 0.1 eq) was added to a solution of rac-3-(((8,8-dimethyl-1,4-dioxaspiro[4.5]decane-7-yl)methyl)amino)-4-nitrobenzonitrile (7 g, 20.27 mmol, 1 eq) in methanol (70 mL). The resulting mixture was stirred overnight at room temperature under a hydrogen atmosphere. The resulting mixture was filtered and the filter cake was washed with methanol (3 x 100 mL). The filtrate was concentrated under reduced pressure to give the crude product rac-4-amino-3-(((8,8-dimethyl-1,4-dioxaspiro[4.5]decane-7-yl)methyl)amino)benzonitrile (6 g crude) as a colorless oil. LCMS (ESI-MS) m / z = 316 [M+H] + .

[0182] rac-1-((8,8-dimethyl-1,4-dioxaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000043.tif Formic acid (0.88 g, 19.02 mmol, 1 eq) was added to a solution of rac-4-amino-3-(((8,8-dimethyl-1,4-dioxaspiro[4.5]decane-7-yl)methyl)amino)benzonitrile (6 g, 19.02 mmol, 1 eq) in trimethoxymethane (58.54 g, 551.64 mmol, 29 eq). The resulting mixture was heated to 80 °C, stirred for 3 h, and concentrated under vacuum to give a crude product. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0% - 100% petroleum ether. Fractions containing the desired mass signal were combined and concentrated under vacuum to give the desired product rac-1-((8,8-dimethyl-1,4-dioxaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (4 g, 60.6% yield over 2 steps) as a white solid. LCMS (ESI-MS) m / z = 326 [M+H] + .

[0183] rac-1-((2,2-Dimethyl-5-oxocyclohexyl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000044.tif53165In formic acid (50 mL), a solution of rac-1-((8,8-dimethyl-1,4-dioxaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (3.80 g, 11.68 mmol, 1 equiv) was stirred at room temperature overnight. The reaction mixture was diluted with saturated sodium bicarbonate solution (100 mL) and extracted with ethyl acetate (3 × 100 mL). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give the crude product rac-1-((2,2-dimethyl-5-oxocyclohexyl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (crude, 3.50 g) as a white solid. LCMS (ESI-MS) m / z = 282 [M+H] + .

[0184] rac-1-(((3S,5R)-6,6-Dimethyl-1-oxaspiro[2.5]octan-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000045.tif53165Sodium hydride (60% in oil, 1.19 g, 49.76 mmol, 4 eq) was added to a solution of trimethylsulfoxonium iodide (10.95 g, 49.76 mmol, 4 eq) in dimethyl sulfoxide (100 mL), and the resulting mixture was stirred at room temperature for 30 minutes. A solution of rac-1-((2,2-dimethyl-5-oxocyclohexyl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (3.50 g, 12.44 mmol, 1 eq) in dimethyl sulfoxide (35 mL) was added dropwise to this mixture. After 8 minutes, the reaction mixture was cooled to 0 °C, quenched slowly with 200 mL of water, and extracted with ethyl acetate (3 × 200 mL). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give the crude product rac-1-(((3S,5R)-6,6-dimethyl-1-oxaspiro[2.5]octan-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (crude, 3 g) as a yellow oil. LCMS (ESI-MS) m / z = 296 [M+H] + .

[0185] rac-1-(((5S,7R)-8,8-dimethyl-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000046.tif58165 Lithium tert-butoxide (1.63 g, 20.31 mmol, 2 equiv) was added to a solution of ethyl carbamate (18.10 g, 203.12 mmol, 20 equiv) in N-methylpyrrolidone (10 mL). After stirring at room temperature for 5 minutes, rac-1-(((3S,5R)-6,6-dimethyl-1-oxaspiro[2.5]octan-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (3 g, 10.16 mmol, 1 equiv) was added dropwise in N-methylpyrrolidone (10 mL). Subsequently, the reaction mixture was heated to 100 °C and stirred overnight. After cooling to room temperature, the reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (3 x 100 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to obtain the crude product. The residue was purified by silica gel column chromatography, eluting with 0% - 2% methanol in dichloromethane, and concentrated under vacuum to give the desired product rac-1-(((5S,7R)-8,8-dimethyl-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (3 g, 75.9% yield over 3 steps) as a white solid. LCMS (ESI-MS) m / z = 339 [M+H] + .

[0186] rac-1-(((5S,7R)-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-8,8-dimethyl-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile Copper(I) iodide (562.78 mg, 2.95 mmol, 1 equiv) was added under a nitrogen atmosphere to a solution of rac-1-(((5S,7R)-8,8-dimethyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (1 g, 2.95 mmol, 1 equiv), 2-(5-chloropyrazin-2-yl)propan-2-ol (0.51 g, 2.95 mmol, 1 equiv), N 1 , N 2 ,N-dimethylethane-1,2-diamine (0.52 g, 5.91 mmol, 2 equiv) and tripotassium phosphate (1.25 g, 5.91 mmol, 2 equiv) in 1,4-dioxane (10 mL). The resulting mixture was heated to 100 °C and stirred overnight. After cooling to room temperature, the resulting mixture was filtered and the filtrate was concentrated under vacuum. The residue was purified by silica gel column chromatography, eluting with methanol in dichloromethane from 0% to 4%. The fractions were combined and concentrated under vacuum to give the desired product rac-1-(((5S,7R)-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-8,8-dimethyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (800 mg, 57.1% yield) as a pale yellow solid. 11H NMR (400 MHz, DMSO-d6) δ 9.10 (d, J = 1.5 Hz, 1H), 8.56 - 8.54 (m, 2H), 8.22 (d, J = 1.4 Hz, 1H), 7.81 (d, J = 8.4 Hz, 1H), 7.59 (dd, J = 8.4, 1.5 Hz, 1H), 5.38 (s, 1H), 4.52 (dd, J = 14.2, 3.7 Hz, 1H), 4.07 (dd, J = 14.3, 11.1 Hz, 1H), 3.86 (d, J = 10.2 Hz, 1H), 3.75 (d, J = 10.2 Hz, 1H), 2.11 (dd, J = 15.1, 12.7 Hz, 1H), 1.87 - 1.64 (m, 4H), 1.42 (d, J = 1.7 Hz, 8H), 1.20 (s, 3H), 1.01 (s, 3H). LCMS (ESI-MS) m / z = 475 [M+H] + .

[0187] 1-(((5S,7R)-3-(5-(2-Hydroxypropan-2-yl)pyrazin-2-yl)-8,8-dimethyl-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile and 1-(((5R,7S)-3-(5-(2-Hydroxypropan-2-yl)pyrazin-2-yl)-8,8-dimethyl-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000048.tif63165rac-1-(((5S,7R)-3-(5-(2-Hydroxypropan-2-yl)pyrazin-2-yl)-8,8-dimethyl-2-oxo-1-oxa-3-azaspiro[4.5]dec-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (800 mg, 1.68 mmol, 1 equiv) was separated by preparative chiral HPLC under the conditions: column: CHIRALPAK IE, 2 * 25 cm, 5 μm, mobile phase A: MtBE (0.5% 2M NH3-MeOH)--HPLC, mobile phase B: EtOH--HPLC, flow rate: 15 mL / min, gradient: 50% B to 50% B in 15 min; wavelength: 220 / 254 nm; RT1 (min): 7.11, RT2 (min): 11.56, sample solvent: EtOH--HPLC, injection volume: 1 mL. The desired fractions were combined and lyophilized to give the product.

[0188] The first eluting isomer: 1-(((5S,7R)-3-(5-(2-Hydroxypropan-2-yl)pyrazin-2-yl)-8,8-dimethyl-2-oxo-1-oxa-3-azaspiro[4.5]dec-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile as a white solid (228.7 mg, purity 96.6%, 100% ee, yield 28.5%). Confirmed by X-ray crystal structure. 1 H NMR (400 MHz, DMSO-d6) δ 9.10 (d, J = 1.5 Hz, 1H), 8.56 - 8.54 (m, 2H), 8.22 (d, J = 1.4 Hz, 1H), 7.81 (d, J = 8.4 Hz, 1H), 7.59 (dd, J = 8.4, 1.5 Hz, 1H), 5.38 (s, 1H), 4.52 (dd, J = 14.2, 3.7 Hz, 1H), 4.07 (dd, J = 14.3), 11.1 Hz, 1H), 3.86 (d, J = 10.2 Hz, 1H), 3.75 (d, J = 10.2 Hz, 1H), 2.11 (dd, J = 15.1, 12.7 Hz, 1H), 1.87 - 1.64 (m, 4H), 1.42 (d, J = 1.7 Hz, 8H), 1.20 (s, 3H), 1.01 (s, 3H). LCMS (ESI-MS) m / z = 475 [M+H] + .

[0189] Second eluting isomer: 1-(((5R,7S)-3-(5-(2-Hydroxypropan-2-yl)pyrazin-2-yl)-8,8-dimethyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile as a white solid (213.1 mg, purity 99.9%, 100% ee, yield 26.6%). 1 H NMR (400 MHz, DMSO-d6) δ 9.10 (d, J = 1.5 Hz, 1H), 8.56 - 8.54 (m, 2H), 8.22 (d, J = 1.4 Hz, 1H), 7.81 (d, J = 8.4 Hz, 1H), 7.59 (dd, J = 8.4, 1.5 Hz, 1H), 5.38 (s, 1H), 4.52 (dd, J = 14.2, 3.7 Hz, 1H), 4.07 (dd, J = 14.3, 11.1 Hz, 1H), 3.86 (d, J = 10.2 Hz, 1H), 3.75 (d, J = 10.2 Hz, 1H), 2.11 (dd, J = 15.1, 12.7 Hz, 1H), 1.87 - 1.64 (m, 4H), 1.42 (d, J = 1.7 Hz, 8H), 1.20 (s, 3H), 1.01 (s, 3H). LCMS (ESI-MS) m / z = 475 [M+H] + .

[0190] Example 2. Preparation of 1-(((5S,7S)-8,8-difluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000049.tif63165

[0191] Reaction Scheme TIFF2025522953000050.tif212165TIFF2025522953000051.tif135165

[0192] Detailed Procedure N, N - Dibenzyl - 1 - methoxymethanamine TIFF2025522953000052.tif27165Potassium carbonate (47.29 g, 342.1 mmol, 1.5 equiv) and sodium sulfate (48.60 g, 342.1 mmol, 1.5 equiv) were added to a mixture of dibenzylamine (45 g, 228.1 mmol, 1 equiv) and paraformaldehyde (6.78 g, 75.27 mmol, 0.3 equiv) in methanol (250 mL). The resulting mixture was stirred at room temperature for 12 h. The resulting mixture was filtered through celite and the filtrate was concentrated under vacuum to give the crude product N,N - dibenzyl - 1 - methoxymethanamine (crude, 50 g) as a yellow oil. 1H NMR (400 MHz, chloroform - d) δ 7.44 - 7.33 (m,9H),7.31 (q,J = 3.1,2.3 Hz,1H),4.08 (s,2H),3.88 (s,4H),3.29 (s, 3H). LCMS (ESI - MS) m / z = 242 [M + H] + .

[0193] rac - 7 - ((Dibenzylamino)methyl)-1,4 - dioxaspiro[4.5]decan - 8 - one TIFF2025522953000053.tif42165(S)-proline (4.77 g, 41.43 mmol, 0.2 eq) was added to a stirred mixture of N,N-dibenzyl-1-methoxymethanamine (50 g, 207.18 mmol, 1 eq) in dimethyl sulfoxide (500 mL), followed by the addition of 1,4-dioxaspiro[4.5]decan-8-one (323.58 g, 2071.82 mmol, 10 eq). The resulting mixture was stirred at 40 °C for 16 h, diluted with water (2000 mL), and extracted with ether (3 × 500 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo to give the crude product. The residue was purified by reverse phase (water containing 0.1% ammonium bicarbonate) in 0% - 50% acetonitrile to afford the desired product rac-7-((dibenzylamino)methyl)-1,4-dioxaspiro[4.5]decan-8-one (30 g, 39.6% yield) as a pale yellow solid. LCMS (ESI-MS) m / z = 366 [M+H] + .

[0194] rac-N,N-dibenzyl-1-(8,8-difluoro-1,4-dioxaspiro[4.5]decan-7-yl)methanamine TIFF2025522953000054.tif37165 Diethylaminosulfur trifluoride (6.62 g, 41.04 mmol, 1.5 equiv) was added dropwise to a stirred mixture of rac-7-((dibenzylamino)methyl)-1,4-dioxaspiro[4.5]decan-8-one (10 g, 27.36 mmol, 1 equiv) in dichloromethane (100 mL) at 0 °C under a nitrogen atmosphere. After the addition was complete, the reaction mixture was stirred at 25 °C for 1 h. The reaction mixture was quenched by the addition of saturated aqueous sodium bicarbonate (200 mL) and extracted with dichloromethane (3 × 150 mL). The combined organic layers were dried over sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0%–10% petroleum ether. The fractions were combined and concentrated under vacuum to give rac-N,N-dibenzyl-1-(8,8-difluoro-1,4-dioxaspiro[4.5]decan-7-yl)methanamine (2.8 g, 26.4% yield) as a colorless oil. LCMS (ESI-MS) m / z = 388 [M+H] + .

[0195] rac-(8,8-Difluoro-1,4-dioxaspiro[4.5]decan-7-yl)methanamine TIFF2025522953000055.tif42165 Palladium on carbon (10%, 1.0 g, 2.52 mmol, 0.35 equiv) was added to a solution of rac-N,N-dibenzyl-1-(8,8-difluoro-1,4-dioxaspiro[4.5]decan-7-yl)methanamine (2.8 g, 7.23 mmol, 1 equiv) in methanol (20 mL) under a nitrogen atmosphere. The reaction mixture was stirred at room temperature overnight under a hydrogen atmosphere. The reaction mixture was filtered and the filter cake was washed with methanol (20 mL). The filtrate was concentrated under reduced pressure to give the crude product rac-(8,8-difluoro-1,4-dioxaspiro[4.5]decan-7-yl)methanamine (crude 6 g) as a colorless oil. LCMS (ESI-MS) m / z = 208 [M+H] + .

[0196] rac-3-(((8,8-difluoro-1,4-dioxaspiro[4.5]decane-7-yl)methyl)amino)-4-nitrobenzonitrile TIFF2025522953000056.tif53165Potassium carbonate (2.4 g, 17.37 mmol, 3 eq) was added to a mixture of rac-(8,8-difluoro-1,4-dioxaspiro[4.5]decane-7-yl)methanamine (1.2 g, 5.79 mmol, 1 eq) and 3-fluoro-4-nitrobenzonitrile (0.96 g, 5.79 mmol, 1 eq) in acetonitrile (15 mL). The resulting mixture was heated to 60 °C, stirred overnight, and quenched by the addition of water (50 mL). The resulting mixture was extracted with ethyl acetate (3 × 50 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo to afford the crude product. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0%–17% petroleum ether. The fractions were combined and concentrated in vacuo to give rac-3-(((8,8-difluoro-1,4-dioxaspiro[4.5]decane-7-yl)methyl)amino)-4-nitrobenzonitrile (840 mg, 32.9% yield over 2 steps) as an orange oil. LCMS (ESI-MS) m / z = 354 [M+H] + .

[0197] rac-4-amino-3-(((8,8-difluoro-1,4-dioxaspiro[4.5]decane-7-yl)methyl)amino)benzonitrile TIFF2025522953000057.tif53165 Iron powder (1.58 g, 28.3 mmol, 10 eq) was added to a mixture of rac-3-(((8,8-difluoro-1,4-dioxaspiro[4.5]decane-7-yl)methyl)amino)-4-nitrobenzonitrile (1 g, 2.83 mmol, 1 eq) and ammonium chloride (0.61 g, 11.3 mmol, 4 eq) in a mixed solvent of water (3 mL) and ethanol (6 mL). The resulting mixture was then stirred at 80 °C for 1 h. After cooling to room temperature, the reaction mixture was filtered and the filter cake was washed with ethanol (30 mL). The filtrate was concentrated under vacuum, the residue was diluted with dichloromethane (30 mL), and washed with water (3 × 30 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to give the crude product rac-4-amino-3-(((8,8-difluoro-1,4-dioxaspiro[4.5]decane-7-yl)methyl)amino)benzonitrile (crude, 700 mg) as a yellow solid. LCMS (ESI-MS) m / z = 324 [M+H] + .

[0198] rac-1-((8,8-difluoro-1,4-dioxaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000058.tif53165 Formic acid (185 mg, 4.02 mmol, 2 equiv) was added to a mixture of rac-4-amino-3-(((8,8-difluoro-1,4-dioxaspiro[4.5]decan-7-yl)methyl)amino)benzonitrile (650 mg, 2.01 mmol, 1 equiv) and triethyl orthoformate (297.92 mg, 2.01 mmol, 1 equiv) in tetrahydrofuran (5 mL). The resulting mixture was heated to 80 °C, stirred for 12 h, and then concentrated under vacuum to give a crude product. The residue was purified by silica gel column chromatography, eluting with methanol in dichloromethane from 0 to 10%. The fractions were combined and concentrated under vacuum to give rac-1-((8,8-difluoro-1,4-dioxaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (600 mg, 63.6% yield over 2 steps) as a yellow solid. LCMS (ESI-MS) m / z = 334 [M+H] + .

[0199] rac-1-((2,2-difluoro-5-oxocyclohexyl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000059.tif53165 A solution of rac-1-((8,8-difluoro-1,4-dioxaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (600 mg, 1.8 mmol, 1 equiv) in formic acid (4 mL) was stirred at 70 °C overnight. The resulting mixture was concentrated under vacuum to give the crude product rac-1-((2,2-difluoro-5-oxocyclohexyl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (crude, 400 mg) as a yellow solid. LCMS (ESI-MS) m / z = 290 [M+H] + .

[0200] rac-1-(((3S,5S)-6,6-difluoro-1-oxaspiro[2.5]octan-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile In dimethyl sulfoxide (5 mL), a stirred mixture of rac-1-((2,2-difluoro-5-oxocyclohexyl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (400 mg, 1.38 mmol, 1 equiv) was heated to 35 °C to dissolve all the solids. Trimethylsulfoxonium iodide (456.45 mg, 2.07 mmol, 1.5 equiv) was added to the above solution, followed by potassium tert-butoxide (232.74 mg, 2.07 mmol, 1.5 equiv). The resulting mixture was stirred at room temperature for 1 h. The reaction mixture was diluted with water (200 mL) and extracted with ethyl acetate (3 × 150 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to give the crude product. The residue was purified by silica gel column chromatography, eluting with methanol in dichloromethane from 0% to 8%. The fractions were combined and concentrated under vacuum to give rac-1-(((3S,5S)-6,6-difluoro-1-oxaspiro[2.5]octan-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (300 mg, yield 71.7%) as a yellow solid. LCMS (ESI-MS) m / z = 304 [M+H] + .

[0201] rac-1-(((5S,7S)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile Lithium tert-butoxide (158.36 mg, 1.98 mmol, 2 equiv) was added to a mixture of rac-1-(((3S,5S)-6,6-difluoro-1-oxaspiro[2.5]octan-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (300 mg, 0.99 mmol, 1 equiv) and ethyl carbamate (132.18 mg, 1.48 mmol, 1.5 equiv) in N-methyl-2-pyrrolidinone (5 mL). The reaction mixture was then heated to 75 °C and stirred overnight. After cooling the reaction mixture to room temperature, the reaction mixture was eluted with water (100 mL) and extracted with ethyl acetate (3 × 100 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to give the crude product. The residue was purified by silica gel column chromatography, eluting with methanol in dichloromethane from 0 to 10%. The fractions were combined and concentrated under vacuum to give rac-1-(((5S,7S)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (300 mg, 87.4% yield) as a yellow solid. LCMS (ESI-MS) m / z = 347 [M+H] + .

[0202] rac-1-(((5S,7S)-8,8-difluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000062.tif68165 Cuprous iodide (198 mg, 1.03 mmol, 1.2 equiv) was added to a mixture of rac-1-(((5S,7S)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]dec-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (300 mg, 0.87 mmol, 1 equiv), 2-(5-chloropyrazin-2-yl)propan-2-ol (149.5 mg, 0.87 mmol, 1 equiv), N 1 , N 2 ,N-dimethylethane-1,2-diamine (152.7 mg, 1.73 mmol, 2 equiv) and tribasic potassium phosphate (441 mg, 2.08 mmol, 2.4 equiv) under a nitrogen atmosphere. The reaction mixture was stirred at 100 °C for 12 h. After cooling to room temperature, the reaction mixture was quenched by the addition of water (20 mL) and extracted with ethyl acetate (3 x 20 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to give the crude product. The residue was purified by silica gel column chromatography, eluting with methanol in dichloromethane from 0 to 10%. The fractions were combined and concentrated under vacuum to give rac-1-(((5S,7S)-8,8-difluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)2-oxo-1-oxa-3-azaspiro[4.5]dec-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (100 mg, 24% yield) as a yellow solid. LCMS (ESI-MS) m / z = 483 [M+H] + .

[0203] 1-(((5R,7R)-8,8-difluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile and 1-(((5S,7S)-8,8-difluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000063.tif73165rac-1-(((5S,7S)-8,8-difluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (150 mg, 0.310 mmol) was separated by preparative chiral-HPLC under the conditions: column: CHIRALPAK ID, 2 * 25 cm, 5 μm, mobile phase A: Hex: DCM = 3:1 (0.5% 2M NH3-methanol)--HPLC, mobile phase B: ethanol-HPLC, flow rate: 20 mL / min, gradient: 45% B~45% B in 17 min; wavelength: 220 nm. The desired fractions were combined and lyophilized to give two products.

[0204] The first eluting isomer: 1-(((5R,7R)-8,8-difluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile as a white solid (26.4 mg, purity 96.2%, 100% ee, yield 17.6%). Confirmed by X-ray structural analysis. 11H NMR (300 MHz, DMSO-d6) δ 9.17 (s, 1H), 8.57 (d, J = 8.0 Hz, 2H), 8.30 (s, 1H), 7.83 (d, J = 8.5 Hz, 1H), 7.62 (s, 1H), 5.39 - 5.37 (m, 1H), 4.66 - 4.64 (m, 1H), 4.38 - 4.36 (m, 1H), 3.93 - 3.91 (m, 2H), 2.90 - 2.88 (m, 1H), 2.28 - 1.78 (m, 6H), 1.44 (s, 6H). LCMS (ESI-MS) m / z = 483 [M+H] + .

[0205] Second eluting isomer: 1 - (((5S,7S)-8,8 - difluoro - 3-(5-(2 - hydroxypropan - 2 - yl)pyrazin - 2 - yl)-2 - oxo - 1 - oxa - 3 - azaspiro[4.5]decan - 7 - yl)methyl)-1H - benzimidazole - 6 - carbonitrile (30.5 mg, 98.7% purity, 97.4% ee, 20.3% yield) was obtained as a white solid. Confirmed by X - ray structural analysis. 1 1H NMR (300 MHz, DMSO-d6) δ 9.17 (s, 1H), 8.71 - 8.55 (m, 2H), 8.30 (s, 1H), 7.83 (d, J = 8.0 Hz, 1H), 7.65 - 7.47 (m, 1H), 5.50 - 5.22 (m, 1H), 4.68 (d, J = 9.6 Hz, 1H), 4.39 - 4.37 (m, 1H), 3.93 - 3.91 (m, 2H), 2.90 - 2.89 (m, 1H), 2.31 - 2.06 (m, 4H), 1.90 - 1.89 (m, 2H), 1.53 - 1.37 (m, 6H). LCMS (ESI-MS) m / z = 483 [M+H] + .

[0206] Example 3. Preparation of 1 - (((4S,6S)-9-(5-(2 - hydroxypropan - 2 - yl)pyrazin - 2 - yl)-8 - oxo - 7 - oxa - 9 - azadispiro[2.2.4 6 .2 3 dodecane - 4 - yl)methyl)-1H - benzimidazole - 6 - carbonitrile TIFF2025522953000064.tif63165

[0207] Reaction Scheme TIFF2025522953000065.tif171165TIFF2025522953000066.tif135165

[0208] Detailed Procedure rac-N,N-Dibenzyl-1-(8-methylene-1,4-dioxaspiro[4.5]decan-7-yl)methanamine TIFF2025522953000067.tif37165 Potassium tert-butoxide (6.1 g, 54.72 mmol, 2 equiv) was added to a mixture of methyltriphenylphosphonium bromide (11.7 g, 32.83 mmol, 1.2 equiv) in toluene (100 mL), followed by the addition of rac-7-((dibenzylamino)methyl)-1,4-dioxaspiro[4.5]decan-8-one (10 g, 27.36 mmol, 1 equiv). The resulting mixture was stirred at 100 °C for 4 h, diluted with water (200 mL), and extracted with dichloromethane (3 × 100 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to give a crude product. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0%–20% petroleum ether. The fractions were combined and concentrated under vacuum to give the desired product rac-N,N-dibenzyl-1-(8-methylene-1,4-dioxaspiro[4.5]decan-7-yl)methanamine (9.2 g, 92.5% yield) as a yellow solid. LCMS (ESI-MS) m / z = 364 [M+H]+.

[0209] rac-N-((7,10-Dioxadispiro[2.2.4 6 .2 3 dodecan-4-yl)methyl)-N-benzyl-1-phenylmethanamine TIFF2025522953000068.tif37165 Methylene iodide (1.77 mL, 22 mmol, 2 eq) and diethylzinc (22.01 mL, 22 mmol, 2 eq) were added to a mixture of rac-N,N-dibenzyl-1-(8-methylen-1,4-dioxaspiro[4.5]decan-7-yl)methanamine (4 g, 11 mmol, 1 eq) in dichloromethane (50 mL). The resulting mixture was stirred at room temperature for 3 h, diluted with saturated aqueous ammonium chloride solution (100 mL), and extracted with dichloromethane (3 × 60 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to give the crude product. The residue was purified by reverse phase using water (containing 0.1% ammonium bicarbonate) in 0% - 40% acetonitrile to give the desired product rac-N-((7,10-dioxadispiro[2.2.4 6 .2 3 dodecane)-4-yl)methyl)-N-benzyl-1-phenylmethanamine (3 g, yield 72.2%) as a yellow solid. LCMS (ESI-MS) m / z = 378 [M+H] + .

[0210] rac-(7,10-dioxadispiro[2.2.4 6 .2 3 dodecane-4-yl)methanamine TIFF2025522953000069.tif37165 Palladium on carbon (10%, 532.77 mg, 5 mmol, 0.6 eq) was added to a mixture of rac-N-((7,10-dioxadispiro[2.2.4 6 .2 3 dodecane-4-yl)methyl)-N-benzyl-1-phenylmethanamine (3 g, 7.94 mmol, 1 eq) in methanol (250 mL). The resulting mixture was stirred at room temperature for 3 h under a hydrogen atmosphere. The mixture was filtered through celite, and the filtrate was concentrated to give the crude product rac-(7,10-dioxadispiro[2.2.4 6 .2 3 dodecane-4-yl)methanamine (crude, 1 g) as a yellow oil. LCMS (ESI-MS) m / z = 198 [M+H]+ .

[0211] rac-3-(((7,10-Dioxadispiro[2.2.4 6 .2 3 dodecane-4-yl)methyl)amino)-4-nitrobenzonitrile TIFF2025522953000070.tif53165Potassium carbonate (1.12 g, 8.11 mmol, 2 eq) was added to a mixture of rac-(7,10-dioxadispiro[2.2.4 6 .2 3 dodecane-4-yl)methanamine (800 mg, 4.05 mmol, 1 eq) and 3-fluoro-4-nitrobenzonitrile (0.67 g, 4.05 mmol, 1 eq) in acetonitrile (20 mL). The resulting mixture was stirred at 50 °C for 4 h, diluted with water (100 mL), and extracted with dichloromethane (3 × 40 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to give the crude product. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0% - 15% petroleum ether. The fractions were combined and concentrated under vacuum to give the desired product rac-3-(((7,10-dioxadispiro[2.2.4 6 .2 3 dodecane-4-yl)methyl)amino)-4-nitrobenzonitrile (1.2 g, 86.3% yield) as a yellow solid. LCMS (ESI-MS) m / z = 344 [M+H] + .

[0212] rac-1-((7,10-Dioxadispiro[2.2.4 6 .2 3 dodecane-4-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000071.tif58165Iron (975.79 mg, 17.47 mmol, 5 eq) was added to a mixed solvent of methanol (60 mL) and ethyl acetate (60 mL), and rac-3-(((7,10-dioxadispiro[2.2.4 6. 2 3((Dodecan-4-yl)methyl)amino)-4-nitrobenzonitrile (1.2 g, 3.49 mmol, 1 equivalent) was added to a mixture of trimethoxymethane (370.85 mg, 3.49 mmol, 1 equivalent) and formic acid (160.84 mg, 3.49 mmol, 1 equivalent). The resulting mixture was stirred at 64 °C for 4 hours and concentrated under vacuum. The residue was diluted with water (50 mL) and extracted with dichloromethane (3 × 50 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to give the crude product. The residue was purified by silica gel column chromatography and eluted with ethyl acetate in 0% - 50% petroleum ether. The fractions were combined and concentrated under vacuum to give rac-1-((7,10-dioxadispiro[2.2.4 6 .2 3 ((Dodecan-4-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (700 mg, yield 62%) was obtained as a yellow solid. LCMS (ESI-MS) m / z = 324 [M+H] + .

[0213] rac-1-((6-oxospiro[2.5]octan-4-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000072.tif53165In formic acid (7 mL), a mixture of rac-1-((7,10-dioxadispiro[2.2.4 6 .2 3 ((Dodecan-4-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (700 mg, 2.16 mmol, 1 equivalent) was heated to 70 °C, stirred overnight, and concentrated under vacuum to give the crude product. The residue was purified by silica gel column chromatography and eluted with ethyl acetate in 0% - 70% petroleum ether. The fractions were combined and concentrated under vacuum to give rac-1-((6-oxospiro[2.5]octan-4-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (500 mg, yield 82.8%) as a yellow solid. LCMS (ESI-MS) m / z = 280 [M+H] + .

[0214] rac-1-(((3S,5S)-1-oxadispiro[2.2.2 6 .2 3 decane-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000073.tif53165Sodium hydroxide (60%, 143.2 mg, 3.58 mmol, 2 equiv) was added to a mixture of rac-1-((6-oxospiro[2.5]octan-4-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (500 mg, 1.79 mmol, 1 equiv) and trimethylsulfoxonium iodide (787 mg, 3.58 mmol, 2 equiv) in dimethyl sulfoxide (2 mL). The resulting mixture was stirred at room temperature for 1 h, diluted with water (40 mL), and extracted with dichloromethane (3 × 30 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to give the crude product rac-1-(((3S,5S)-1-oxadispiro[2.2.2 6 .2 3 decane-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (crude, 470 mg) as a white solid. LCMS (ESI-MS) m / z = 294 [M+H] + .

[0215] rac-1-(((4S,6S)-8-oxo-7-oxa-9-azadispiro[2.2.4 6 .2 3 dodecane-4-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000074.tif58165A mixture of lithium tert-butoxide (64.13 mg, 0.8 mmol, 0.5 equiv) and ethyl carbamate (713 mg, 8 mmol, 5 equiv) in N-methyl-2-pyrrolidone (11.75 mL) was stirred at room temperature for 10 min, followed by rac-1-(((3S,5S)-1-oxadispiro[2.2.2 6 .2 3(Decan-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (470 mg, 1.60 mmol, 1 equiv) was added. The resulting mixture was stirred at 75 °C for 4 h. After cooling to room temperature, the resulting mixture was diluted with water (30 mL) and extracted with dichloromethane (3 x 30 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give the crude product rac-1-(((4S,6S)-8-oxo-7-oxa-9-azadispiro[2.2.4 6 .2 3 (dodecane-4-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (crude, 420 mg) as a white solid. LCMS (ESI-MS) m / z = 337 [M+H] +

[0216] rac-1-(((4S,6S)-9-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-8-oxo-7-oxa-9-azadispiro[2.2.4 6 .2 3 (dodecane-4-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000075.tif68165Copper(I) iodide (113.23 mg, 0.59 mmol, 0.5 equiv) was added to rac-1-(((4S,6S)-8-oxo-7-oxa-9-azadispiro[2.2.4 6 .2 3 (dodecane-4-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (400 mg, 1.19 mmol, 1 equiv), 2-(5-chloropyrazin-2-yl)propan-2-ol (205.25 mg, 1.19 mmol, 1 equiv), N 1 ,N 2It was added to a mixture of -dimethylethane-1,2-diamine (209.64 mg, 2.38 mmol, 2 eq) and tribasic potassium phosphate (504.80 mg, 2.38 mmol, 2 eq). The resulting mixture was stirred at 100 °C for 16 h under a nitrogen atmosphere. After cooling to room temperature, the reaction mixture was diluted with water (100 mL) and extracted with dichloromethane (3 x 100 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to obtain the crude product. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0% - 60% petroleum ether. The fractions were combined and concentrated under vacuum to give the desired product rac-1-(((4S,6S)-9-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-8-oxo-7-oxa-9-azadispiro[2.2.4 6 .2 3 dodecan-4-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (400 mg, yield 71.3%) as a white solid. LCMS (ESI-MS) m / z = 473 [M+H] + .

[0217] 1-(((4R,6R)-9-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-8-oxo-7-oxa-9-azadispiro[2.2.4 6 .2 3 dodecan-4-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile and 1-(((4S,6S)-9-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-8-oxo-7-oxa-9-azadispiro[2.2.4 6 . 2 3 dodecan-4-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000076.tif73165rac-1-(((4S,6S)-9-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-8-oxo-7-oxa-9-azadispiro-[2.2.4 6 .2 3((4R,6R)-9-(5-(2-Hydroxypropan-2-yl)pyrazin-2-yl)-8-oxo-7-oxa-9-azadispiro[2.2.4.2]dodecan-4-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (400 mg, 0.84 mmol, 1 equiv), a racemic compound, was separated by preparative chiral HPLC under the following conditions: column: CHIRALPAK IF, 2 * 25 cm, 5 μm; mobile phase A: MtBE (0.5% 2M NH3-MeOH)--HPLC, mobile phase B: EtOH--HPLC; flow rate: 15 mL / min; gradient: 30% B to 30% B in 17 min; wavelength: 220 / 254 nm; RT1 (min): 10.68; RT2 (min): 13.184; sample solvent: EtOH--HPLC; injection volume: 0.4 mL. The desired fractions were combined and lyophilized to give two products.

[0218] The first eluting isomer: 1-(((4R,6R)-9-(5-(2-Hydroxypropan-2-yl)pyrazin-2-yl)-8-oxo-7-oxa-9-azadispiro[2.2.4 6 .2 3 dodecan-4-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (173.6 mg, purity 99.1%, 100.0% ee). 1 H NMR (300 MHz, DMSO-d6) δ 9.26 (d, J = 1.5 Hz, 1H), 8.60 (d, J = 1.5 Hz, 1H), 8.51 (s, 1H), 8.28 (d, J = 1.5 Hz, 1H), 7.82 (d, J = 8.4 Hz, 1H), 7.58 (dd, J = 8.4, 1.5 Hz, 1H), 5.44 (s, 1H), 4.54 (dt, J = 16.2, 8.1 Hz, 2H), 3.91 (q, J = 10.3 Hz, 2H), 2.49 - 2.40 (m, 1H), 2.30 - 2.19 (m, 1H), 2.14 - 2.00 (m, 1H), 1.93 - 1.72 (m, 4H), 1.46 (s, 6H), 0.95 - 0.82 (m, 1H), 0.41 - 0.17 (m, 3H). LCMS (ESI-MS) m / z = 473 [M + H] + .

[0219] Second eluting isomer: As a white solid, 1-(((4S,6S)-9-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-8-oxo-7-oxa-9-azadispiro 6 .2 3 dodecan-4-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (159.8 mg, purity 99.1%, 99.0% ee). Absolute configuration confirmed by X-ray crystal structure. 1 H NMR (300 MHz, DMSO-d6) δ 9.26 (d, J = 1.5 Hz, 1H), 8.60 (d, J = 1.5 Hz, 1H), 8.50 (s, 1H), 8.28 (d, J = 1.5 Hz, 1H), 7.82 (d, J = 8.4 Hz, 1H), 7.58 (dd, J = 8.4, 1.5 Hz, 1H), 5.43 (s, 1H), 4.52 (t, J = 8.0 Hz, 2H), 3.91 (q, J = 10.3 Hz, 2H), 2.49 - 2.38 (m, 1H), 2.33 - 2.20 (m, 1H), 2.15 - 2.01 (m, 1H), 1.93 - 1.72 (m, 4H), 1.45 (s, 6H), 0.97 - 0.80 (m, 1H), 0.38 - 0.20 (m, 3H). LCMS (ESI-MS) m / z = 473 [M+H] + .

[0220] Example 4. Preparation of 1-(((5S,7S)-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2,8-dioxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000077.tif63165

[0221] Reaction scheme TIFF2025522953000078.tif167165TIFF2025522953000079.tif223165

[0222] Detailed procedure 1-Methoxy-5-methylcyclohexa-1,4-diene TIFF2025522953000080.tif22165Ammonia (1200 mL) was concentrated in an empty, oven-dried three-necked round-bottom flask under an inert atmosphere of nitrogen at -78 °C, equipped with a cold finger containing dry ice-acetone. To this was added 1-methoxy-3-methylbenzene (64 g, 524.59 mmol, 1 equiv) in diethyl ether (600 mL), followed by tert-butanol (420 mL). Then, lithium wire (36.41 g, 5245.9 mmol, 10 equiv) was carefully added. After the addition, the reaction mixture was warmed to -33 °C, after which the reaction mixture turned dark blue. The reaction contents were stirred at -33 °C for 3 h, after which ammonium chloride (14.03 g, 2622.95 mmol, 5 equiv) was added. The reaction mixture was then removed from the -33 °C bath, the cold finger was removed, and the reaction contents (exposed to air) were warmed to room temperature and stirred for an additional 14 h. The resulting organic material was dissolved in 1000 mL of pentane and washed with water (3 × 1000 mL). The organic matter was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the crude product 1-methoxy-5-methylcyclohexa-1,4-diene (crude, 60 g) as a yellow oil. LCMS (ESI-MS) m / z = 125 [M+H] + .

[0223] 3-Methylcyclohex-3-en-1-one TIFF2025522953000081.tif22165Oxalic acid (6.53 g, 72.58 mmol, 0.15 equiv) was added to a solution of 1-methoxy-5-methylcyclohexa-1,4-diene (60 g, 483.87 mmol, 1 equiv) in methanol (400 mL) and water (100 mL). The resulting solution was stirred at room temperature for 16 h. The residue was concentrated under reduced pressure to give the crude product 3-methylcyclohex-3-en-1-one (crude, 50 g) as a white solid. LCMS (ESI-MS) m / z = 111 [M+H] + .

[0224] 1-Methyl-7-oxabicyclo[4.1.0]heptan-3-one TIFF2025522953000082.tif22165In 500 mL of dichloromethane, 3-chloroperbenzoic acid (101.97 g, 590.90 mmol, 1.3 equiv) was added to a solution of 3-methylcyclohex-3-en-1-one (50 g, 454.54 mmol, 1 equiv) at 0 °C. The reaction mixture was stirred at 0 °C for 3 h, and the reaction was quenched with 500 mL of saturated aqueous sodium thiosulfate solution and stirred for 5 min. The layers were separated, and the aqueous layer was extracted with dichloromethane (2 x 500 mL). The combined organics were washed with saturated aqueous sodium bicarbonate solution (3 × 500 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give the crude product 1-methyl-7-oxabicyclo[4.1.0]heptan-3-one (crude, 54 g) as a pale yellow oil. 1 H NMR (400 MHz, chloroform-d) δ 3.23 (d, J = 2.9 Hz, 1H), 2.80 (d, J = 19.0 Hz, 1H), 2.59 (dd, J = 19.0, 0.9 Hz, 1H), 2.43 - 2.33 (m, 2H), 2.23 - 2.14 (m, 2H), 1.38 (s, 3H). LCMS (ESI-MS) m / z = 127 [M+H] + .

[0225] 4-((tert-Butyldiphenylsilyl)oxy)-3-methylcyclohex-2-en-1-one TIFF2025522953000083.tif32165tert-Butyldiphenylsilyl chloride (117.80 g, 428.57 mmol, 1 equiv) was added to a solution of 1-methyl-7-oxabicyclo[4.1.0]heptan-3-one (54 g, 428.57 mmol, 1 equiv), diisopropylethylamine (110.78 g, 857.14 mmol, 2 equiv) and 4-dimethylaminopyridine (10.47 g, 85.71 mmol, 0.2 equiv) in dichloromethane (600 mL) under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 72 h. The reaction mixture was poured into 500 mL of dichloromethane and 500 mL of water, and the layers were separated. The organic layer was washed, dried over anhydrous sodium sulfate, filtered, concentrated to give a crude product. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0% - 10% petroleum ether. Fractions with the desired mass signal were combined and concentrated under vacuum to give the desired product 4-((tert-butyldiphenylsilyl)oxy)-3-methylcyclohex-2-en-1-one (60 g, 31.3% yield over 4 steps) as a pale yellow oil. LCMS (ESI-MS) m / z = 365 [M+H] + .

[0226] rac-(3S,4R)-4-((tert-Butyldiphenylsilyl)oxy)-3-methyl-3-(nitromethyl)cyclohexan-1-one TIFF2025522953000084.tif42165 Nitromethane (4.01 g, 65.75 mmol, 1.2 equiv) was added to a solution of 4-((tert-butyldiphenylsilyl)oxy)-3-methylcyclohex-2-en-1-one (20 g, 54.94 mmol, 1 equiv) and benzyltrimethylammonium hydroxide (13.75 g, 82.41 mmol, 1.5 equiv) in methanol (100 mL). The resulting mixture was stirred at room temperature overnight and concentrated under vacuum to give a crude product. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0% - 10% petroleum ether. The fractions were combined and concentrated under vacuum to give the desired product rac-(3S,4R)-4-((tert-butyldiphenylsilyl)oxy)-3-methyl-3-(nitromethyl)cyclohexan-1-one (18 g, 77.1% yield) as a pale yellow oil. 1H NMR (400 MHz, DMSO-d6) δ 7.62 - 7.49 (m, 4H), 7.42 - 7.36 (m, 6H), 4.41 - 4.33 (m, 2H), 3.65 - 3.59 (m, 1H), 2.34 - 1.99 (m, 6H), 1.09 - 0.98 (m, 12H). LCMS (ESI-MS) m / z = 426 [M+H] + .

[0227] rac-tert-Butyl (((7S,8R)-7-methyl-7-(nitromethyl)-1,4-dioxaspiro[4.5]dec-8-yl)oxy)diphenylsilane TIFF2025522953000085.tif42165Ethylene glycol (4.33 g, 69.72 mmol, 1.5 equiv) was added to a solution of rac-(3S,4R)-4-((tert-butyldiphenylsilyl)oxy)-3-methyl-3-(nitromethyl)cyclohexan-1-one (19.8 g, 46.58 mmol, 1 equiv) and trimethoxymethane (7.40 g, 69.72 mmol, 1.5 equiv) in dichloromethane (100 mL). The resulting reaction mixture was stirred at room temperature for 5 minutes and then cooled to 0 °C in an ice bath. Methanesulfonic acid (0.67 g, 6.97 mmol, 0.15 equiv) was added dropwise to this mixture. The resulting mixture was removed from the bath and warmed to room temperature and stirred for 2 hours. The reaction mixture was concentrated under vacuum. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0% - 10% petroleum ether. The fractions were combined and concentrated under vacuum to give the desired product rac-tert-butyl (((7S,8R)-7-methyl-7-(nitromethyl)-1,4-dioxaspiro[4.5]decan-8-yl)oxy)diphenylsilane (18.5 g, 84.7% yield) as a pale yellow oil. 1 HNMR (400 MHz,DMSO-d6) δ 7.62-7.49 (m,4H),7.42-7.36 (m,6H),4.41-4.33 (m,2H),3.50-3.34 (m,4H),3.25 -3.19 (m, 1H), 1.65-1.49 (m, 6H), 1.09 (s, 9H), 0.98 (s, 3H). LCMS (ESI-MS) m / z = 470 [M+H] + .

[0228] rac-((7S,8R)-8-((tert-butyldiphenylsilyl)oxy)-7-methyl-1,4-dioxaspiro[4.5]decan-7-yl)methanamine TIFF2025522953000086.tif42165 Iron powder (21.75 g, 389.4 mmol, 10 equivalents) was added to a solution of rac-tert-butyl(((7S,8R)-7-methyl-7-(nitromethyl)-1,4-dioxaspiro[4.5]decan-8-yl)oxy)diphenylsilane (18.3 g, 39.01 mmol, 1 equivalent) and ammonium chloride (8.33 g, 155.76 mmol, 4 equivalents) in a mixed solvent of ethanol (120 mL) and water (40 mL). The resulting mixture was heated to 80 °C and stirred for 2 h. After cooling to room temperature, the resulting mixture was filtered and the filter cake was washed with ethanol (2 x 200 mL). The filtrate was concentrated under reduced pressure to give the crude product rac-((7S,8R)-8-((tert-butyldiphenylsilyl)oxy)-7-methyl-1,4-dioxaspiro[4.5]decan-7-yl)methanamine (crude, 17 g) as a yellow solid. LCMS (ESI-MS) m / z = 440 [M+H] + .

[0229] rac-3-((((7S,8R)-8-((tert-butyldiphenylsilyl)oxy)-7-methyl-1,4-dioxaspiro[4.5]decan-7-yl)methyl)amino)-4-nitrobenzonitrile TIFF2025522953000087.tif48165 Potassium carbonate (10.68 g, 77.3 mmol, 2 equiv) was added to a solution of rac-((7S,8R)-8-((tert-butyldiphenylsilyl)oxy)-7-methyl-1,4-dioxaspiro[4.5]decan-7-yl)methanamine (17 g, 38.7 mmol, 1 equiv) and 3-fluoro-4-nitrobenzonitrile (6.41 g, 38.7 mmol, 1 equiv) in acetonitrile (150 mL). The resulting mixture was heated to 40 °C and stirred overnight. After cooling to room temperature, the reaction mixture was diluted with water (200 mL) and extracted with ethyl acetate (3 x 200 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to give the crude product. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0% - 15% petroleum ether. Fractions with the desired mass signal were combined and concentrated under vacuum to give the desired product rac-3-((((7S,8R)-8-((tert-butyldiphenylsilyl)oxy)-7-methyl-1,4-dioxaspiro[4.5]decan-7-yl)methyl)amino)-4-nitrobenzonitrile (14 g, 61.3% yield over 2 steps) as an orange solid. 1H NMR (400 MHz, DMSO-d6) δ 8.30 - 8.26 (m, 1H), 7.61 - 7.48 (m, 4H), 7.41 - 7.34 (m, 7H), 7.15 (dd, J = 8.2, 4.6 Hz), 1H), 6.74 (s, 1H), 3.59 - 3.57 (m, 4H), 3.44 - 3.34 (m, 1H), 3.02 - 2.99 (m, 1H), 2.88 - 2.85 (m, 1H), 1.85 - 1.30 (m, 6H), 1.07 (s, 9H), 0.94 (s, 3H). LCMS (ESI-MS) m / z = 586 [M+H] + .

[0230] rac-4-Amino-3-((((7S,8R)-8-((tert-butyldiphenylsilyl)oxy)-7-methyl-1,4-dioxaspiro[4.5]decan-7-yl)methyl)amino)benzonitrile TIFF2025522953000088.tif48165 Iron powder (13.34 g, 238.91 mmol, 10 eq) was added to a solution of rac-3-((((7S,8R)-8-((tert-butyldiphenylsilyl)oxy)-7-methyl-1,4-dioxaspiro[4.5]decan-7-yl)methyl)amino)-4-nitrobenzonitrile (14 g, 23.93 mmol, 1 eq) and ammonium chloride (5.11 g, 95.56 mmol, 4 eq) in a mixed solvent of ethanol (120 mL) and water (40 mL). The resulting mixture was heated to 80 °C and stirred for 2 h. After cooling to room temperature, the resulting mixture was filtered and the filter cake was washed with ethanol (2 x 200 mL). The filtrate was concentrated under reduced pressure to give the crude product rac-4-amino-3-((((7S,8R)-8-((tert-butyldiphenylsilyl)oxy)-7-methyl-1,4-dioxaspiro[4.5]decan-7-yl)methyl)amino)benzonitrile (crude, 13 g) as a yellow solid. LCMS (ESI-MS) m / z = 556 [M+H] + .

[0231] rac-1-(((7S,8R)-8-((tert-butyldiphenylsilyl)oxy)-7-methyl-1,4-dioxaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000089.tif58165Formic acid (1.08 g, 23.38 mmol, 1 equiv) was added to a solution of rac-4-amino-3-((((7S,8R)-8-((tert-butyldiphenylsilyl)oxy)-7-1,4-dioxaspiro[4.5]decan-7-yl)methyl)amino)benzonitrile (13 g, 23.4 mmol, 1 equiv) in trimethoxymethane (71.95 g, 678 mmol, 29 equiv). The resulting mixture was heated to 80 °C, stirred for 3 h, and concentrated under vacuum to afford the crude product. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0%–100% petroleum ether. Fractions with the desired mass signal were combined and concentrated under vacuum to give the desired product rac-1-(((7S,8R)-8-((tert-butyldiphenylsilyl)oxy)-7-methyl-1,4-dioxaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (10 g, 73.9% yield over 2 steps) as a pink solid. 1 H NMR (400 MHz, DMSO-d6) δ 8.40–8.35 (m, 1H), 7.98–7.93 (m, 1H), 7.82–7.79 (m, 1H), 7.63–7.51 (m, 5H), 7.36 –7.29 (m, 6H), 3.59–3.46 (m, 6H), 3.44–3.34 (m, 1H), 1.59–1.43 (m, 6H), 1.01 (s, 9H), 0.98 (s, 3H). LCMS (ESI-MS) m / z = 566 [M+H] + .

[0232] rac-1-(((1S,2R)-2-((tert-butyldiphenylsilyl)oxy)-1-methyl-5-oxocyclohexyl)methyl)-1H-benzo[d]imidazole-6-carbonitrile In formic acid (100 mL), a solution of rac-1-(((7S,8R)-8-((tert-butyldiphenylsilyl)oxy)-7-methyl-1,4-dioxaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (10 g, 17.7 mmol, 1 equiv) was stirred at 70 °C overnight. The reaction mixture was diluted with saturated sodium bicarbonate solution (100 mL) and extracted with ethyl acetate (3 × 100 mL). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give the crude product rac-1-(((1S,2R)-2-((tert-butyldiphenylsilyl)oxy)-1-methyl-5-oxocyclohexyl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (crude, 10 g) as a yellow solid. LCMS (ESI-MS) m / z = 522 [M+H] + .

[0233] rac-1-(((3S,5S,6R)-6-((tert-butyldiphenylsilyl)oxy)-5-methyl-1-oxaspiro[2.5]octan-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000091.tif53165Sodium hydride (60% in mineral oil, 3.07 g, 76.6 mmol, 4 equiv) was added to a solution of trimethylsulfoxonium iodide (16.87 g, 76.6 mmol, 4 equiv) in dimethyl sulfoxide (100 mL), and the resulting mixture was stirred at room temperature for 30 minutes. To this mixture was added dropwise rac-1-(((1S,2R)-2-((tert-butyldiphenylsilyl)oxy)-1-methyl-5-oxocyclohexyl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (10 g, 19.19 mmol, 1 equiv) in dimethyl sulfoxide (100 mL). After 8 minutes, the reaction mixture was cooled to 0 °C, quenched slowly with 200 mL of water, and extracted with ethyl acetate (3 × 200 mL). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give the crude product rac-1-(((3S,5S,6R)-6-((tert-butyldiphenylsilyl)oxy-5-methyl-1-oxaspiro[2.5]octan-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (crude, 10 g) as a yellow solid. LCMS (ESI-MS) m / z = 536 [M+H] + .

[0234] rac-1-(((5S,7S,8R)-8-((tert-butyldiphenylsilyl)oxy)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile Lithium tert-butoxide (2.99 g, 37.32 mmol, 2 equivalents) was added to a solution of ethyl carbamate (33.21 g, 373.20 mmol, 20 equivalents) in N-methylpyrrolidone (100 mL). After stirring at room temperature for 5 minutes, a solution of rac-1-(((3S,5S,6R)-6-((tert-butyldiphenylsilyl)oxy)-5-methyl-1-oxaspiro[2.5]octan-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (10 g, 18.69 mmol, 1 equivalent) in N-methylpyrrolidone (100 mL) was added dropwise. Subsequently, the reaction mixture was heated to 100 °C and stirred overnight. After cooling to room temperature, the reaction mixture was diluted with water (200 mL) and extracted with ethyl acetate (3 x 200 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to obtain the crude product. The residue was purified by silica gel column chromatography, eluting with methanol in dichloromethane from 0 to 2%. The fractions with the desired mass signal were combined and concentrated under vacuum to obtain the desired product rac-1-(((5S,7S,8R)-8-((tert-butyldiphenylsilyl)oxy)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (5.5 g, 53.8% yield over 3 steps) as a pale pink solid. 1 H NMR (400 MHz, DMSO-d6) δ 8.40 - 8.35 (m, 1H), 7.98 - 7.93 (m, 1H), 7.82 - 7.79 (m, 1H), 7.63 - 7.51 (m, 5H), 7.36 - 7.29 (m, 6H), 6.50 (s, 1H), 3.59 - 3.46 (m, 2H), 3.44 - 3.34 (m, 1H), 3.25 - 3.20 (m, 2H), 1.79 - 1.45 (m, 6H), 1.22 (s, 9H), 0.97 (s, 3H). LCMS (ESI-MS) m / z = 579 [M+H] + .

[0235] rac-1-(((5S,7S,8R)-8-Hydroxy-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000093.tif63165rac-1-(((5S,7S,8R)-8-((tert-Butyldiphenylsilyl)oxy)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (579 mg, 1 mmol, 1 equiv) was added to a solution of tetrabutylammonium fluoride in tetrahydrofuran (1 M, 1 mL, 1 mmol, 1 equiv). The mixture was stirred overnight at room temperature. The next day, calcium carbonate (200.18 mg, 2 mmol, 2 equiv) and DOWEX 50WX8-400 (800 mg, 1 equiv) were added to the reaction mixture, followed by methanol (2 mL). The resulting mixture was stirred at room temperature for 1 h, filtered, and the filter cake was washed with methanol (2 x 10 mL). The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with methanol in 0 - 10% dichloromethane. Fractions having the desired mass signal were combined and concentrated under vacuum to give the desired product rac-1-(((5S,7S,8R)-8-hydroxy-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (270 mg, 79.4% yield) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 8.30 (s, 1H), 7.98 - 7.93 (m, 1H), 7.82 - 7.79 (m, 1H), 7.63 - 7.51 (m, 1H), 6.45 (s, 1H), 4.75 (s, 1H), 3.63 - 3.59 (m, 2H), 3.47 - 3.35 (m, 1H), 3.25 - 3.20 (m, 2H), 1.81 - 1.47 (m, 6H), 0.97 (s, 3H). LCMS (ESI-MS) m / z = 341 [M+H] + .

[0236] rac-1-(((5S,7S)-7-methyl-2,8-dioxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000094.tif63165 Dess-Martin periodinane (224.45 mg, 0.53 mmol, 1.2 equiv) was added to a solution of rac-1-(((5S,7S,8R)-8-hydroxy-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (150 mg, 0.44 mmol, 1 equiv) in dichloromethane (2 mL). The resulting mixture was stirred at room temperature for 3 h. The residue was purified by silica gel column chromatography, eluting with methanol in 0 - 2% dichloromethane. Fractions containing the desired mass signal were combined and concentrated in vacuo to give the desired product rac-1-((((5S,7S)-7-methyl-2,8-dioxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (130 mg, 87.18% yield) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 8.30 (s, 1H), 7.98 - 7.93 (m, 1H), 7.82 - 7.79 (m, 1H), 7.63 - 7.51 (m, 1H), 6.45 (s, 1H), 3.63 - 3.59 (m, 2H), 3.25 - 3.20 (m, 2H), 2.35 - 2.29 (m, 2H), 2.11 - 1.85 (m, 4H), 1.12 (s, 3H). LCMS (ESI-MS) m / z = 339 [M+H] + .

[0237] rac-1-(((5S,7S)-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2,8-dioxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile Copper(I) iodide (72.37 mg, 0.38 mmol, 1 equiv) was added under a nitrogen atmosphere to a solution of rac-1-(((5S,7S)-7-methyl-2,8-dioxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (130 mg, 0.38 mmol, 1 equiv), 2-(5-chloropyrazin-2-yl)propan-2-ol (65.74 mg, 0.38 mmol, 1 equiv), N 1 ,N 2 -dimethylethane-1,2-diamine (66.88 mg, 0.76 mmol, 2 equiv) and tripotassium phosphate (161.32 mg, 0.76 mmol, 2 equiv) in 1,4-dioxane (2 mL). The resulting mixture was heated to 100 °C and stirred overnight. After cooling to room temperature, the resulting mixture was filtered and the filtrate was concentrated under vacuum. The residue was purified by silica gel column chromatography, eluting with methanol in 0 - 3% dichloromethane. Fractions having the desired mass signal were combined and concentrated under vacuum to give the desired product rac-1-(((5S,7S)-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2,8-dioxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (100 mg, 54.7% yield) as a pale yellow solid. 11H NMR (300 MHz, DMSO-d6) δ 9.27 (s, 1H), 8.64 (d, J = 4.0 Hz, 1H), 8.32 (d, J = 10.1 Hz, 2H), 7.86‐7.79 (m, 1H), 7.60 (t, J = 5.6 Hz, 1H), 5.44 (d, J = 3.3 Hz, 1H), 4.85 (d, J = 14.7 Hz, 1H), 4.60 (d, J = 14.9 Hz, 1H), 4.16 (d, J = 10.1 Hz, 1H), 4.05 (d, J = 10.5 Hz, 1H), 3.19 (s, 1H), 2.58 (d, J = 12.4 Hz, 2H), 2.46 ‐2.19 (m, 3H), 1.47 (t, J = 3.2 Hz, 6H), 0.95 (d, J = 3.6 Hz, 3H). LCMS (ESI-MS) m / z = 475 [M+H] + .

[0238] 1-(((5S,7S)-3-(5-(2-Hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2,8-dioxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile and 1-(((5R,7R)-3-(5-(2-Hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl 2,8-dioxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000096.tif68165rac-1-(((5S,7S)-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2,8-dioxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (100 mg, 0.21 mmol, 1 equiv) was separated by preparative chiral HPLC under the conditions: CHIRALPAK ID, 2*25 cm, 5 μm, mobile phase A: EtOH--HPLC, mobile phase B: MtBE (0.5% 2M NH3-MeOH)--HPLC, flow rate: 13 mL / min, gradient: 50% B to 50% B in 13 min, wavelength: 220 / 254 nm; RT1 (min): 8.31, RT2 (min): 23.51, sample solvent: EtOH:DCM = 1:1--HPLC, injection volume: 2 mL. The desired fractions were combined and lyophilized to give the product.

[0239] The first eluting isomer: 1-(((5S,7S)-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2,8-dioxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (25.6 mg, purity 98.8%, 98.5% ee, yield 25.6%) was obtained as a white solid. 11H NMR (300 MHz, DMSO-d6) δ 9.27 (s, 1H), 8.64 (d, J = 4.0 Hz, 1H), 8.32 (d, J = 10.1 Hz, 2H), 7.86 - 7.79 (m, 1H), 7.60 (t, J = 5.6 Hz, 1H), 5.44 (d, J = 3.3 Hz, 1H), 4.85 (d, J = 14.7 Hz, 1H), 4.60 (d, J = 14.9 Hz, 1H), 4.16 (d, J = 10.1 Hz, 1H), 4.05 (d, J = 10.5 Hz, 1H), 3.19 (s, 1H), 2.58 (d, J = 12.4 Hz, 2H), 2.46 - 2.19 (m, 3H), 1.47 (t, J = 3.2 Hz, 6H), 0.95 (d, J = 3.6 Hz, 3H). LCMS (ESI-MS) m / z = 475 [M+H] + .

[0240] Second eluting isomer: 1-(((5R,7R)-3-(5-(2-Hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2,8-dioxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (22.7 mg, purity 99.6%, 100% ee, yield 22.7%) was obtained as a white solid. 11H NMR (300 MHz, DMSO-d6) δ 9.27 (s, 1H), 8.64 (d, J = 4.0 Hz, 1H), 8.32 (d, J = 10.1 Hz, 2H), 7.86 - 7.79 (m, 1H), 7.60 (t, J = 5.6 Hz, 1H), 5.44 (d, J = 3.3 Hz, 1H), 4.85 (d, J = 14.7 Hz, 1H), 4.60 (d, J = 14.9 Hz, 1H), 4.16 (d, J = 10.1 Hz, 1H), 4.05 (d, J = 10.5 Hz, 1H), 3.19 (s, 1H), 2.58 (d, J = 12.4 Hz, 2H), 2.46 - 2.19 (m, 3H), 1.47 (t, J = 3.2 Hz, 6H), 0.95 (d, J = 3.6 Hz, 3H). LCMS (ESI-MS) m / z = 475 [M+H] + .

[0241] Example 5. Preparation of 1-(((5S,7S,8R)-8-fluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000097.tif63165

[0242] Reaction scheme TIFF2025522953000098.tif198165TIFF2025522953000099.tif104165

[0243] Detailed procedure rac-Ethyl 8-oxo-1,4-dioxaspiro[4.5]decan-7-carboxylate In TIFF2025522953000100.tif48165, a mixture of diethyl carbonate (94 g, 79.6 mmol) and sodium hydride (60% in mineral oil, 37.5 g, 93.8 mmol) in tetrahydrofuran (800 mL) was heated to reflux, and then a solution of 1,4-dioxaspiro[4.5]decan-8-one (50.0 g, 32.0 mmol) in tetrahydrofuran (230 mL) was added. The resulting mixture was stirred for 3 hours and then cooled to room temperature. Water (500 mL) and acetic acid (500 mL) were added to quench the reaction. The resulting solution was then extracted with ethyl acetate (3 × 500 mL), the organic layer was dried over sodium sulfate, filtered, and concentrated to give rac-ethyl 8-oxo-1,4-dioxaspiro[4.5]decan-7-carboxylate (52 g, 0.23 mol) as a yellow oil. This was used without further purification. LCMS (ESI-MS) m / z = 229.1 [M+H]+.

[0244] rac-ethyl 7-methyl-8-oxo-1,4-dioxaspiro[4.5]decan-7-carboxylate In TIFF2025522953000101.tif42165, iodomethane (48.5 g, 341 mmol) was added to a mixture of rac-ethyl 8-oxo-1,4-dioxaspiro[4.5]decan-7-carboxylate (52 g, 228 mmol) and cesium carbonate (149 g, 456 mmol) in acetonitrile (600 mL). The resulting mixture was stirred at room temperature for 4 hours. The mixture was filtered and the filtrate was concentrated to give a crude product. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0% - 30% petroleum ether to give the product rac-ethyl 7-methyl-8-oxo-1,4-dioxaspiro[4.5]decan-7-carboxylate (45 g, 81.5%) as a pale yellow oil. LCMS (ESI-MS) m / z = 243.1 [M+H] + .

[0245] rac-ethyl 8-fluoro-7-methyl-1,4-dioxaspiro[4.5]deca-8-ene-7-carboxylate In dichloromethane (500 mL), to a mixture of rac-ethyl 7-methyl-8-oxo-1,4-dioxaspiro[4.5]decane-7-carboxylate (30 g, 124 mmol), diethylaminosulfur trifluoride (39.9 g, 248 mmol) was added at 0 °C. The solution was warmed to room temperature and stirred for 5 h. Then the mixture was slowly added to the stirred aqueous solution, sodium bicarbonate (600 mL) was added at 0 °C, and the resulting solution was extracted with dichloromethane (3 × 300 mL). The organic layer was dried over sodium sulfate, filtered, and concentrated to give rac-ethyl 8-fluoro-7-methyl-1,4-dioxaspiro[4.5]dec-8-ene-7-carboxylate (crude, 35 g). LCMS (ESI-MS) m / z = 245.1 [M+H] + .

[0246] rac-(8-Fluoro-7-methyl-1,4-dioxaspiro[4.5]dec-8-en-7-yl)methanol In tetrahydrofuran (600 mL), to a mixture of rac-ethyl 8-fluoro-7-methyl-1,4-dioxaspiro[4.5]dec-8-ene-7-carboxylate (35 g, 143 mmol), lithium aluminum hydride (115 mL, 286 mmol) was added at 0 °C, the mixture was stirred at room temperature for 2 h, and then the mixture was quenched with cold ethanol. The resulting mixture was filtered and the filtrate was concentrated to give the crude product rac-(8-fluoro-7-methyl-1,4-dioxaspiro[4.5]dec-8-en-7-yl)methanol (crude, 30 g). LCMS (ESI-MS) m / z = 203.1 [M+H] + .

[0247] rac-2-((8-Fluoro-7-methyl-1,4-dioxaspiro[4.5]dec-8-en-7-yl)methyl)isoindoline-1,3-dione In tetrahydrofuran (300 mL), to a mixture of rac-(8-fluoro-7-methyl-1,4-dioxaspiro[4.5]deca-8-en-7-yl)methanol (25 g, 124 mmol), isatin (20 g, 136 mmol) and triphenyl phosphate (48.6 g, 185 mmol), diisopropyl azodicarboxylate (37.5 g, 185 mmol) was added. The mixture was stirred at room temperature overnight, and the next morning, the mixture was poured into ice water (200 mL). The resulting solution was extracted with ethyl acetate (3 × 200 mL), the organic layer was dried over sodium sulfate, filtered and concentrated to give the crude product. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0% - 40% petroleum ether to give the product rac-2-((8-fluoro-7-methyl-1,4-dioxaspiro[4.5])deca-8-en-7-yl)methyl)isoindoline-1,3-dione (18 g, 43.9%) as a yellow oil. LCMS (ESI-MS) m / z = 332.1 [M+H] + .

[0248] rac-(8-fluoro-7-methyl-1,4-dioxaspiro[4.5]deca-8-en-7-yl)methanamine In ethanol (100 mL), to a mixture of rac -2-((8-fluoro-7-methyl-1,4-dioxaspiro[4.5]deca-8-en-7-yl)methyl)isoindoline-1,3-dione (18 g, 54 mmol), hydrazine (17 g, 272 mmol, 80%) was added. The mixture was stirred at room temperature overnight and then diluted with water (300 mL). The resulting solution was extracted with ethyl acetate (3 × 200 mL), the organic layer was dried over sodium sulfate, filtered and concentrated to give the product rac-(8-fluoro-7-methyl-1,4-dioxaspiro)[4.5]deca-8-en-7-yl)methanamine (crude, 8 g). LCMS (ESI-MS) m / z = 202.1 [M+H] + .

[0249] rac-((7S,8R)-8-Fluoro-7-methyl-1,4-dioxaspiro[4.5]decane-7-yl)methanamine TIFF2025522953000106.tif37165In methanol (80 mL), palladium carbon (4 g, 50 wt%, 10% on carbon) was added to a mixture of rac-(8-fluoro-7-methyl-1,4-dioxaspiro[4.5]dec-8-en-7-yl)methanamine (8 g, 39.8 mmol). The mixture was stirred overnight at room temperature under a hydrogen atmosphere (2 atm). The suspension was filtered and the filtrate was concentrated under vacuum to give rac-((7S,8R)-8-fluoro-7-methyl-1,4-dioxaspiro[4.5]decane-7-yl)methanamine (7.2 g, crude) as a yellow oil. LCMS (ESI-MS) m / z = 204.1 [M+H] + .

[0250] rac-3-((((7S,8R)-8-Fluoro-7-methyl-1,4-dioxaspiro[4.5]decane-7-yl)methyl)amino)-4-nitrobenzonitrile In acetonitrile (100 mL), to a mixture of rac-((7S,8R)-8-fluoro-7-methyl-1,4-dioxaspiro[4.5]decan-7-yl)methanamine (7.2 g, 35.4 mmol) and 3-fluoro-4-nitrobenzonitrile (5.88 g, 35.4 mmol) was added potassium carbonate (9.79 g, 70.8 mmol), and the resulting mixture was stirred at 50 °C for 4 hours. The suspension was filtered, the filtrate was diluted with water (200 mL), and then the resulting solution was extracted with dichloromethane (3 × 200 mL). The organic layer was dried over sodium sulfate, filtered, and concentrated to obtain the crude product. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0% - 15% petroleum ether to give the product rac-3-((((7S,8R)-8-fluoro-7-methyl)-1,4-dioxaspiro[4.5]decan-7-yl)methyl)amino)-4-nitrobenzonitrile (6 g, 48.4%) as an orange solid. LCMS (ESI-MS) m / z = 350.1 [M+H] + .

[0251] rac-4-Amino-3-((((7S,8R)-8-fluoro-7-methyl-1,4-dioxaspiro[4.5]decan-7-yl)methyl)amino)benzonitrile In ethanol (100 mL) and water (30 mL), to a mixture of rac-3-((((7S,8R)-8-fluoro-7-methyl-1,4-dioxaspiro[4.5]decan-7-yl)methyl)amino)-4-nitrobenzonitrile (5.5 g, 15.7 mmol) and ammonium chloride (4.21 g, 78.7 mmol) was added iron (4.40 g, 78.7 mmol), and the mixture was stirred at 80 °C for 2 hours. The mixture was cooled to room temperature and filtered. The filtrate was concentrated under vacuum to give rac-4-amino-3-((((7S,8R)-8-fluoro-7-methyl-1,4-dioxaspiro[4.5]decan-7-yl)methyl)amino)benzonitrile (crude, 4.2 g) as a yellow solid. LCMS(ESI-MS) m / z = 320.2 [M+H]+ .

[0252] rac-1-(((7S,8R)-8-Fluoro-7-methyl-1,4-dioxaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile In TIFF2025522953000109.tif53165, formic acid (0.91 g, 19.7 mmol) was added to a mixture of rac-4-amino-3-((((7S,8R)-8-fluoro-7-methyl-1,4-dioxaspiro[4.5]decane-7-yl)methyl)amino)benzonitrile (4.2 g, 13.2 mmol) and trimethoxymethane (2.09 g, 19.7 mmol) in acetonitrile (20 mL). The mixture was stirred at 80 °C for 2 hours and then cooled to room temperature. The resulting solution was concentrated under vacuum to give the crude product. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0% - 45% petroleum ether, to give the product rac-1-(((7S,8R)-8-fluoro-7-methyl-1,4-dioxaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (4 g, 92.3%) as a yellow solid. LCMS (ESI-MS) m / z = 330.2 [M+H] + .

[0253] rac-1-(((1S,2R)-2-Fluoro-1-methyl-5-oxocyclohexyl)methyl)-1H-benzo[d]imidazole-6-carbonitrile 15 mL of formic acid was added to rac-1-(((7S,8R)-8-fluoro-7-methyl-1,4-dioxaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (4 g, 12.1 mmol). The mixture was stirred at 70 °C overnight and concentrated to afford the product rac-1-(((1S,2R)-2-fluoro-1-methyl-5-oxocyclohexyl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (crude, 3.6 g) as a yellow oil. LCMS (ESI-MS) m / z = 286.1 [M+H] + .

[0254] rac-1-(((3S,5S,6R)-6-fluoro-5-methyl-1-oxaspiro[2.5]octan-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile and rac-1-(((3R,5S,6R)-6-fluoro-5-methyl-1-oxaspiro[2.5]octan-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000111.tif53165To a mixture of DMSO (50 mL) and trimethylsulfoxonium iodide (5.24 g, 23.8 mmol) was added sodium hydride (60% in mineral oil, 1.43 g, 35.7 mmol). The mixture was stirred at 10 °C for 10 minutes, then rac-1-(((1S,2R)-2-fluoro-1-methyl-5-oxocyclohexyl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (3.4 g, 11.9 mmol) was added. Subsequently, the mixture was stirred at room temperature for 10 minutes and then quenched with aqueous ammonium chloride solution (200 mL). The resulting solution was extracted with ethyl acetate (3 × 100 mL), the organic layer was dried over sodium sulfate, filtered, and concentrated to obtain a crude product. To separate the diastereomers, the residue was purified by reverse-phase chromatography, eluting with acetonitrile in water from 0% to 45% to give the more polar rac-1-(((3R,5S,6S)-6-fluoro-5-methyl-1-oxaspiro[2.5]octan-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (600 mg) as a pale yellow solid. Further elution with acetonitrile in water (55%) gave the less polar rac-1-(((3S,5S,6R)-6-fluoro-5-methyl-1-oxaspiro[2.5]octan-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (1.2 g) as a pale yellow solid. LCMS (ESI-MS) m / z = 300.1 [M+H] + .

[0255] rac-1-(((5S,7S,8R)-8-fluoro-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000112.tif58165 In N-methylpyrrolidone (10 mL), a mixture of ethyl carbamate (1.49 g, 16.7 mmol) and lithium tert-butoxide (401 mg, 5.01 mmol) was stirred at room temperature for 10 minutes, then rac-1-(((3S,5S,6R)-6-fluoro-5-methyl-1-oxaspiro[2.5]octan-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (500 mg, 1.67 mmol) was added, and the mixture was stirred at 100 °C overnight. The next day, the mixture was quenched with water (60 mL) and extracted with ethyl acetate (3 x 30 mL). The combined organic layers were dried over sodium sulfate, filtered, concentrated to give the crude product. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0% to 100% petroleum ether to give the product rac-1-(((5S,7S,8R)-8-fluoro-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (400 mg) as a colorless oil. LCMS (ESI-MS) m / z = 343.1 [M+H] + .

[0256] rac-1-(((5S,7S,8R)-8-fluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000113.tif68165 Copper(I) iodide (110.46 mg, 0.58 mmol, 1 equiv) was added to rac-1-(((5S,7S,8R)-8-fluoro-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (200 mg, 0.58 mmol, 1 equiv), 2-(5-chloropyrazin-2-yl)propan-2-ol (100.34 mg, 0.58 mmol, 1 equiv) in 1,4-dioxane (2 mL), N 1 ,N2 It was added to a solution of -dimethylethane-1,2-diamine (102.08 mg, 1.16 mmol, 2 equivalents) and tripotassium phosphate (246.23 mg, 1.16 mmol, 2 equivalents) under a nitrogen atmosphere. The resulting mixture was heated to 100 °C and stirred overnight. After cooling to room temperature, the resulting mixture was filtered and the filtrate was concentrated under vacuum. The residue was purified by silica gel column chromatography and eluted with methanol in 0 - 3% dichloromethane. Fractions having the desired mass signal were combined and concentrated under vacuum to give the desired product rac-1-(((5S,7S,8R)-8-fluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (200 mg, yield 71.43%) as a pale yellow solid. 1 H NMR (400 MHz, DMSO-d6) δ 9.18 (d, J = 1.5 Hz, 1H), 8.59 (d, J = 1.5 Hz, 1H), 8.45 (s, 1H), 8.26 (d, J = 1.5 Hz, 1H), 7.84 (d, J = 8.4 Hz, 1H), 7.60 (dd, J = 8.3, 1.5 Hz, 1H), 5.42 (s, 1H), 4.67 - 4.50 (m, 1H), 4.38 - 4.25 (m, 2H), 3.83 (d, J = 2.0 Hz, 2H), 2.12 - 1.99 (m, 2H), 1.97 - 1.88 (m, 2H), 1.80 - 1.69 (m, 2H), 1.44 (d, J = 2.3 Hz, 6H), 1.19 (s, 3H). LCMS (ESI-MS) m / z = 479 [M+H] + .

[0257] 1-(((5S,7S,8R)-8-Fluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile and 1-(((5R,7R,8S)-8-fluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000114.tif63165 rac-1-(((5S,7S,8R)-8-Fluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyll-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (200 mg, 0.42 mmol, 1 equivalent) was separated by preparative chiral HPLC under the conditions: CHIRALPAK IF, 2 * 25 cm, 5 μm, mobile phase A: MtBE (0.5% 2M NH3-MeOH)--HPLC, mobile phase B: EtOH--HPLC, flow rate: 14 mL / min, gradient: 30% B - 30% B in 29 min, wavelength: 220 / 254 nm, RT1 (min): 13.529; RT2 (min): 21.063, sample solvent: EtOH--HPLC, injection volume: 0.8 mL. The desired fractions were combined and lyophilized to obtain the product.

[0258] Example 5A (the first eluting isomer): 1-(((5S,7S,8R)-8-Fluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2 -oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile as a white solid (77.3 mg, purity 97.0%, 100% ee, yield 38.65%). 11H NMR (400 MHz, DMSO-d6) δ 9.18 (d, J = 1.5 Hz, 1H), 8.59 (d, J = 1.5 Hz, 1H), 8.45 (s, 1H), 8.26 (d, J = 1.5 Hz, 1H), 7.84 (d, J = 8.4 Hz, 1H), 7.60 (dd, J = 8.3, 1.5 Hz, 1H), 5.42 (s, 1H), 4.67 - 4.50 (m, 1H), 4.38 - 4.25 (m, 2H), 3.83 (d, J = 2.0 Hz, 2H), 2.12 - 1.99 (m, 2H), 1.97 - 1.88 (m, 2H), 1.80 - 1.69 (m, 2H), 1.44 (d, J = 2.3 Hz, 6H), 1.19 (s, 3H). LCMS (ESI-MS) m / z = 479 [M+H] + .

[0259] Example 5B (the second eluting isomer): 1-(((5R,7R,8S)-8-Fluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile as a white solid (76.5 mg, purity 99.5%, 100% ee, yield 38.25%). 1 1H NMR (400 MHz, DMSO-d6) δ 9.18 (d, J = 1.5 Hz, 1H), 8.59 (d, J = 1.5 Hz, 1H), 8.45 (s, 1H), 8.26 (d, J = 1.5 Hz, 1H), 7.84 (d, J = 8.4 Hz, 1H), 7.60 (dd, J = 8.3, 1.5 Hz, 1H), 5.42 (s, 1H), 4.67 - 4.50 (m, 1H), 4.38 - 4.25 (m, 2H), 3.83 (d, J = 2.0 Hz, 2H), 2.12 - 1.99 (m, 2H), 1.97 - 1.88 (m, 2H), 1.80 - 1.69 (m, 2H), 1.44 (d, J = 2.3 Hz, 6H), 1.19 (s, 3H). LCMS (ESI-MS) m / z = 479 [M+H]+ . The structure was confirmed by X-ray crystallographic analysis.

[0260] Example 6. Preparation of 1-(((4R,6S)-9-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-4-methyl-8-oxo-7-oxa-9-azadispiro[2.2.4 6 .2 3 dodecan-4-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000115.tif53165

[0261] Detailed procedure rac-N,N-dibenzyl-1-(7-methyl-8-methylene-1,4-dioxaspiro[4.5]decan-7-yl)methanamine In toluene (600 mL), to a mixture of methyltriphenylphosphonium bromide (70.6 g, 198 mmol), potassium 2-methylpropane-2-olate (22.2 g, 198 mmol) was added and the mixture was stirred at room temperature for 10 minutes, then rac-7-((dibenzylamino)methyl)-7-methyl-1,4-dioxaspiro[4.5]decan-8-one (50 g, 132 mmol) was added. The mixture was stirred at 100 °C for 4 hours and then cooled to room temperature. The mixture was quenched with water (600 mL) and the resulting solution was extracted with ethyl acetate (3 x 400 mL). The organic layer was dried over sodium sulfate, filtered and concentrated to give a crude product. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0% - 10% petroleum ether to give rac-N,N-dibenzyl-1-(7-methyl-8-methylene-1,4-dioxaspiro[4.5]decan-7-yl)methanamine (45 g) as a colorless oil. LCMS (ESI-MS) m / z = 378.2 [M+H] + .

[0262] rac-N,N-dibenzyl-1-(4-methyl-7,10-dioxadispiro[2.2.4 6 .23 (4-Methyldodecan-4-yl)methanamine TIFF2025522953000117.tif37165 To a mixture of rac-N,N-dibenzyl-1-(7-methyl-8-methylidene-1,4-dioxaspiro[4.5]decan-7-yl)methanamine (45 g, 119 mmol) in diethyl ether (600 mL) was added diethylzinc (1 M in THF, 358 mL, 358 mmol) at 0 °C. The mixture was stirred for 30 minutes, then diiodomethane (95.8 g, 358 mmol) was added and the resulting solution was warmed to room temperature and stirred overnight. The mixture was quenched slowly with aqueous ammonium chloride solution (600 mL), then the resulting solution was extracted with DCM (3 × 200 mL), the organic layer was dried over sodium sulfate, filtered and concentrated to give the crude product. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in petroleum ether containing 0% - 10% ethyl acetate to give rac-N,N-dibenzyl-1-(4-methyl-7,10-dioxadispiro[2.2.4 6 .2 3 (4-Methyldodecan-4-yl)methanamine (5 g) was obtained as a colorless oil. LCMS (ESI-MS) m / z = 392.3 [M+H] + .

[0263] rac-(4-Methyl-7,10-dioxadispiro[2.2.4 6 .2 3 (4-Methyldodecan-4-yl)methanamine TIFF2025522953000118.tif37165 To a solution of rac-N,N-dibenzyl-1-(4-methyl-7,10-dioxadispiro[2.2.4 6 .2 3 (4-Methyldodecan-4-yl)methanamine (4 g, 10.2 mmol) in methanol (100 mL) was added palladium (2 g, 50 wt%, 10% on carbon). The suspension was stirred under a hydrogen atmosphere for 3 hours, then filtered. The filtrate was concentrated to give the crude product rac-(4-methyl-7,10-dioxadispiro[2.2.4 6 .2 3(4-Dodecyl)methanamine (2 g crude) was obtained as a yellow oil. LCMS (ESI-MS) m / z = 212.2 [M+H] + .

[0264] rac-3-(((4-Methyl-7,10-dioxadispiro[2.2.4 6 .2 3 (4-Dodecyl)methyl)amino)-4-nitrobenzonitrile TIFF2025522953000119.tif53165 In acetonitrile (30 mL), a mixture of rac-(4-methyl-7,10-dioxadispiro[2.2.4 6 .2 3 (4-Dodecyl)methanamine (2.2 g, 10.4 mmol) and 3-fluoro-4-nitrobenzonitrile (1.73 g, 10.4 mmol) was added potassium carbonate (4.32 g, 31.2 mmol). The mixture was stirred at 50 °C for 4 h and then cooled to room temperature. The mixture was filtered and the filtrate was concentrated to give a crude product. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0% - 18% petroleum ether to give rac-3-(((4-methyl-7,10-dioxadispiro[2.2.4 6 .2 3 (4-Dodecyl)methyl)amino)-4-nitrobenzonitrile (3.7 g, 99.4%) as an orange solid. LCMS (ESI-MS) m / z = 358.2 [M+H] + .

[0265] rac-4-Amino-3-(((4-methyl-7,10-dioxadispiro[2.2.4 6 .2 3 (4-Dodecyl)methyl)amino)-benzonitrile TIFF2025522953000120.tif53165 In ethanol (50 mL), rac-3-(((4-methyl-7,10-dioxadispiro[2.2.4 6 .2 3((4-Methyldodecan-4-yl)methyl)amino)-4-nitrobenzonitrile (3.7 g, 10.4 mmol) and ammonium chloride (2.77 g, 51.8 mmol) were added to a mixture of iron (2.89 g, 51.8 mmol). The mixture was stirred at 70 °C for 2 h and then cooled to room temperature. The mixture was then filtered, and the filtrate was diluted with water (50 mL), extracted with ethyl acetate (3 × 30 mL), dried over sodium sulfate, filtered, and concentrated to give the product rac-4-amino-3-(((4-methyl-7,10-dioxadispiro[2.2.4 6 .2 3 ((4-Methyldodecan-4-yl)methyl)aminobenzonitrile (3.3 g, 97.3%) was obtained as a yellow solid. LCMS (ESI-MS) m / z = 328.2 [M+H] + .

[0266] rac-1-((4-Methyl-7,10-dioxadispiro[2.2.4 6 .2 3 ((4-Methyldodecan-4-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000121.tif53165In acetonitrile (30 mL), to a mixture of rac-4-amino-3-(((4-methyl-7,10-dioxadispiro[2.2.4 6 .2 3 ((4-Methyldodecan-4-yl)methyl)aminobenzonitrile (3.2 g, 9.77 mmol) were added trimethoxymethane (1.56 g, 14.7 mmol) and formic acid (0.67 g, 14.7 mmol). The resulting mixture was stirred at 80 °C for 2 h and then cooled to room temperature. The mixture was concentrated to give a crude product, and the residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0% - 30% petroleum ether to give rac-1-((4-methyl-7,10-dioxadispiro[2.2.4 6 .2 3 ((4-Methyldodecan-4-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (3 g, 90.9%) was obtained as a yellow solid. LCMS (ESI-MS) m / z = 338.2 [M+H] + .

[0267] rac-1-((4-methyl-6-oxospiro[2.5]octan-4-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000122.tif53165 Formic acid (20 mL) was added to rac-1-((4-methyl-7,10-dioxadispiro[2.2.4 6 .2 3 dodecan-4-yl)methyl)-1H-benzo[d]imidazole- 6-carbonitrile (3 g, 8.89 mmol), and the mixture was stirred at 70 °C overnight and then cooled to room temperature. The solution was concentrated to give the product rac-1-((4-methyl-6-oxospiro[2.5]octan-4-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (2.3 g) as a yellow oil. LCMS (ESI-MS) m / z = 294.2 [M+H] + .

[0268] 1-(5-methyl-1-oxadispiro[2.2.26.23]decane-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile In methyl sulfoxide (30 mL), potassium tert-butoxide (0.99 g, 8.86 mmol) was added to a mixture of rac-1-((4-methyl-6-oxospiro[2.5]octan-4-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (2 g, 6.82 mmol) and trimethylsulfoxonium iodide (1.95 g, 8.86 mmol). The mixture was stirred at room temperature for 10 minutes and then quenched with aqueous ammonium chloride solution (50 mL), and the resulting solution was extracted with dichloromethane (3 × 30 mL). The combined organic layers were dried over sodium sulfate, filtered, concentrated to give a crude product, and the residue was purified by silica gel column chromatography, eluting with ethyl acetate in petroleum ether containing 0% - 80% ethyl acetate to give 1-(((5-methyl-1-oxadispiro[2.2.26.23]decane-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (1.7 g) as a white solid. LCMS (ESI-MS) m / z = 308.2 [M+H] + .

[0269] 1-((6-Hydroxy-6-(hydroxymethyl)-4-methylspiro[2.5]octan-4-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000124.tif48165In N,N-dimethylformamide (10 mL) and water (10 mL), trifluoroacetic acid (1.26 g, 11.1 mmol) was added to a mixture of 1-((5-methyl-1-oxadispiro[2.2.2[2.3]decane-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (1.7 g, 5.53 mmol). The mixture was stirred at room temperature overnight and diluted with aqueous sodium bicarbonate solution (50 mL). The resulting solution was extracted with dichloromethane (3 × 30 mL), dried over sodium sulfate, filtered, and concentrated to give the crude product 1-((6-hydroxy-6-(hydroxymethyl)-4-methylspiro[2.5]octan-4-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (2.02 g) as a yellow oil. LCMS (ESI-MS) m / z = 326.2 [M+H] + .

[0270] rac-1-(((3S,5R)-5-methyl-1-oxadispiro[2.2.2 6 .2 3 decane-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile and rac-1-(((3R,5R)-5-methyl-1-oxadispiro[2.2.2 6 .2 3 decane-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000125.tif53165 In dichloromethane (30 mL), to a mixture of rac-1-((6-hydroxy-6-(hydroxymethyl)-4-methylspiro[2.5]octan-4-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (2.02 g, 6.21 mmol), p-toluenesulfonyl chloride (1.78 g, 9.31 mmol), 4-dimethylaminopyridine (0.38 g, 3.10 mmol) and triethylamine (1.88 g, 18.6 mmol) were added. The mixture was stirred at room temperature for 6 hours and then quenched with water (50 mL). The resulting solution was extracted with dichloromethane (3 × 30 mL), the organic layer was dried over sodium sulfate, filtered and concentrated to obtain a crude intermediate. The residue was dissolved in acetonitrile (20 mL), potassium carbonate (1.72 g, 12.4 mmol) was added, the mixture was stirred at 50 °C for 3 hours, filtered, and the filtrate was concentrated to obtain a crude product. The residue was purified by preparative TLC (EA), Rf = 0.5 to give the product rac-1-(((3S,5R)-5-methyl-1-oxadispiro[2.2.2 6 .2 3 decane-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (210 mg, 11%) as a white solid and Rf = 0.3, and rac-1-(((3R,5R)-5-methyl- 1-oxadispiro[2.2.2 6 .2 3 decane-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (350 mg, 18.3%) was obtained as a white solid. LCMS (ESI-MS) m / z = 308.2 [M+H] + .

[0271] rac-1-(((4R,6S)-4-methyl-8-oxo-7-oxa-9-azadispiro[2.2.4 6 .2 3 dodecane-4-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000126.tifIn 581651 - methyl - 2 - pyrrolidinone (10 mL), a mixture of ethyl carbamate (1.01 g, 11.4 mmol) and lithium tert - butoxide (182 mg, 2.28 mmol) was stirred at room temperature for 10 minutes, then rac - 1 - (((3S,5R) - 5 - methyl - 1 - oxadispiro[2.2.2 6 .2 3 decane - 5 - yl)methyl) - 1H - benzo[d]imidazole - 6 - carbonitrile (350 mg, 1.14 mmol) was added. The mixture was stirred at 150 °C for 6 hours and then cooled to room temperature. The mixture was quenched with water (50 mL) and extracted with ethyl acetate (3 × 30 mL). The organic layer was dried over sodium sulfate, filtered, and concentrated to obtain the crude product. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0% - 100% petroleum ether to give rac - 1 - (((4R,6S) - 4 - methyl - 8 - oxo - 7 - oxa - 9 - azadispiro[2.2.4 6 .2 3 dodecane - 4 - yl)methyl) - 1H - benzo[d]imidazole - 6 - carbonitrile (330 mg, 82.7%) as a yellow solid. LCMS (ESI - MS) m / z = 351.2 [M + H] + .

[0272] rac - 1 - (((4R,6S) - 9 - (5 - (2 - hydroxypropan - 2 - yl)pyrazin - 2 - yl) - 4 - methyl - 8 - oxo - 7 - oxa - 9 - azadispiro[2.2.4 6 .2 3 dodecane - 4 - yl)methyl) - 1H - benzo[d]imidazole - 6 - carbonitrile TIFF2025522953000127.tifIn 1,4 - dioxane (15 mL), rac - 1 - (((4R,6S) - 4 - methyl - 8 - oxo - 7 - oxa - 9 - azadispiro[2.2.4 6 .2 3((4-(Dodecyl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (330 mg, 0.94 mmol), 2-(5-chloropyrazin-2-yl)propan-2-ol (244 mg, 1.41 mmol), N 1 ,N 2 -Dimethylethane-1,2-diamine (165 mg, 1.88 mmol), tribasic potassium phosphate (399 mg, 1.88 mmol), and copper(I) iodide (90 mg, 0.47 mmol) were added. The mixture was stirred at 100 °C for 16 h under a nitrogen atmosphere and then cooled to room temperature. The mixture was diluted with water (50 mL), extracted with ethyl acetate (3 × 30 mL), the organic layer was dried over sodium sulfate, filtered, and concentrated to give a crude product. The residue was purified by silica gel column chromatography, eluting with 0%–7% methanol in dichloromethane to give rac-1-(((4R,6S)-9-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-4-methyl-8-oxo-7-oxa-9-azadispiro[2.2.4 6 .2 3 ((4-(Dodecyl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (350 mg) as a white solid. LCMS (ESI-MS) m / z = 487.2 [M+H] + .

[0273] 1-(((4R,6S)-9-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-4-methyl-8-oxo-7-oxa-9-azadispiro[2.2.4 6 .2 3 ((4-(Dodecyl)methyl)-1H-benzo[d]imidazole-6-carbonitrile and 1-(((4S,6R)-9-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-4-methyl-8-oxo-7-oxa-9-azadispiro[2.2.4 6 .2 3 ((4-(Dodecyl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000128.tif58165rac-1-(((4R,6S)-9-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-4-methyl-8-oxo-7-oxa-9-azadispiro[2.2.4 6 .2 3 dodecane-4-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (350 mg, 0.72 mmol) was separated by preparative chiral HPLC under the following conditions: CHIRALPAK ID, 2 x 25 cm, 5 μm, mobile phase A: MTBE (0.5% 2M NH3-MeOH)--HPLC, mobile phase B: EtOH--HPLC, flow rate: 12 mL / min, gradient: 20% B to 20% B in 25 minutes, wavelength: 220 / 254 nm, RT1 (min): 11.542, sample solvent: EtOH--HPLC, injection volume: 0.7 mL. The desired fractions were combined and lyophilized to obtain two products.

[0274] The first eluting isomer: 1-(((4R,6S)-9-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-4-methyl-8-oxo-7-oxa-9-azadispiro[2.2.4 6 .2 3 dodecane-4-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (113.6 mg, purity 95.3%, 100% ee). 1HNMR (400 MHz, DMSO-d6) δ 9.17 (d, J = 1.5 Hz, 1H), 8.60 (d, J = 1.5 Hz, 1H), 8.44 (s, 1H), 8.36 (d, J = 1.5 Hz, 1H), 7.81 (d, J = 8.4 Hz, 1H), 7.57 (dd, J = 8.4, 1.5 Hz, 1H), 5.41 (s, 1H), 4.20 - 4.06 (m, 3H), 3.95 (dd, J = 8.4, 1.5 Hz, 1H) d, J = 10.2 Hz, 1H), 2.17 (d, J = 13.9 Hz, 1H), 1.97 - 1.74 (m, 4H), 1.52 - 1.34 (m, 7H), 0.91 (s, 3H), 0.81 - 0.79 (m, 1H), 0.59 - 0.49 (m, 1H), 0.31 - 0.17 (m, 2H). LCMS (ESI-MS) m / z = 487.2 [M+H] + .

[0275] Second eluting isomer: 1-(((4S,6R)-9-(5-(2-Hydroxypropan-2-yl)pyrazin-2-yl)-4-methyl-8-oxo-7-oxa-9-azaspiro[2.2.4 6 .2 3 dodecan-4-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (111.0 mg, purity 99.5%, 100% ee). 11H NMR (400 MHz, DMSO-d6) δ 9.17 (d, J = 1.5 Hz, 1H), 8.60 (d, J = 1.6 Hz, 1H), 8.44 (s, 1H), 8.38-8.33 (m, 1H), 7.81 (d, J = 8.4 Hz, 1H), 7.57 (dd, J = 8.4, 1.4 Hz, 1H), 5.41 (s, 1H), 4.20-4.04 (m, 3H), 3.95 (d, J = 10.2 Hz, 1H), 2.17 (d, J = 13.9 Hz, 1H), 2.01-1.71 (m, 4H), 1.54-1.37 (m, 7H), 0.91 (s, 3H), 0.837-0.77(m, 1H), 0.59-0.47(m, 1H), 0.32-0.17 (m, 2H). LCMS (ESI-MS) m / z = 487.2 [M+H] + .

[0276] Example 7. Preparation of 1-(((5S,7R)-7-(fluoromethyl)-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000129.tif53165

[0277] Detailed procedure (1,4-Dioxaspiro[4.5]decan-6-en-7-yl)methanol In 37165 tetrahydrofuran (400 mL), a solution of methyl 1,4-dioxaspiro[4.5]dec-6-ene-7-carboxylate (19.5 g, 98.4 mmol) and lithium aluminum hydride (3.73 g, 98.8 mmol) was stirred at 0 °C for 1 hour. The resulting mixture was quenched with aqueous sodium hydroxide solution, and then the resulting mixture was filtered. The filter cake was washed with tetrahydrofuran (2 × 100 mL). The filtrate was concentrated under reduced pressure to obtain the crude product, (1,4-dioxaspiro[4.5]dec-6-en-7-yl)methanol (7 g), as a yellow oil. 1 H NMR (400 MHz, DMSO-d6) δ 5.48 (s, 1H), 3.92 - 3.79 (m, 6H), 1.92 - 1.83 (m, 2H), 1.71 - 1.61 (m, 4H).

[0278] 7-(Fluoromethyl)-1,4-dioxaspiro[4.5]dec-6-ene In 27165 dichloromethane (140 mL), a solution of (1,4-dioxaspiro[4.5]dec-6-en-7-yl)methanol (7.0 g, 41.1 mmol) and diethylaminosulfur trifluoride (7.95 g, 49.3 mmol) was stirred at room temperature overnight. The mixture was quenched with aqueous sodium carbonate solution (200 mL), and then the resulting mixture was extracted with dichloromethane (3 × 100 mL). The organic layer was dried, filtered, and concentrated to obtain a crude product. The residue was purified by silica gel column chromatography, eluting with dichloromethane (100%) to obtain 7-(fluoromethyl)-1,4-dioxaspiro[4.5]dec-6-ene (6.0 g) as a brown oil. 1 H NMR (400 MHz, DMSO-d6) δ 5.90 (s, 1H), 5.08 (d, J = 4.0 Hz, 1H), 2.52 - 2.48 (m, 1H), 2.42 - 2.16 (m, 5H), 2.08 - 1.84 (m, 3H), 1.73 - 1.59 (m, 1H), 1.44 - 1.31 (m, 1H).

[0279] 3-(Fluoromethyl)cyclohex-2-en-1-one TIFF2025522953000132.tif27165 A solution of 7-(fluoromethyl)-1,4-dioxaspiro[4.5]dec-6-ene (6.0 g, 34.8 mmol) in formic acid (10 mL) was stirred at 80 °C for 1 hour. The resulting mixture was concentrated under vacuum. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0% - 10% petroleum ether to give 3-(fluoromethyl)cyclohex-2-en-1-one (2.0 g) as a colorless oil.

[0280] rac-3-(Fluoromethyl)-3-(nitromethyl)cyclohexan-1-one TIFF2025522953000133.tif32165 A solution of 3-(fluoromethyl)cyclohex-2-en-1-one (2.0 g, 15.6 mmol) and nitromethane (1.14 g, 18.7 mmol) in benzyltrimethylazanium methoxide (3 mL) was stirred at room temperature overnight. The resulting mixture was concentrated under vacuum. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0% - 20% petroleum ether to give rac-3-(fluoromethyl)-3-(nitromethyl)cyclohexan-1-one (1.0 g) as a colorless oil. 1 HNMR (400 MHz, DMSO-d6) δ 4.69 - 4.30 (m, 4H), 2.45 - 2.21 (m, 4H), 1.95 - 1.70 (m, 4H).

[0281] rac-7-(Fluoromethyl)-7-(nitromethyl)-1,4-dioxaspiro[4.5]decane In dichloromethane (20 mL), a solution of rac-3-(fluoromethyl)-3-(nitromethyl)cyclohexan-1-one (1.0 g, 5.3 mmol), ethylene glycol (0.49 g, 7.9 mmol), and trimethoxymethane (0.84 g, 7.9 mmol) was stirred at room temperature for 5 minutes. The solution was cooled to 0 °C, and methanesulfonic acid (80 mg, 0.79 mmol) was added. The resulting mixture was warmed to room temperature and stirred overnight. The solution was concentrated under vacuum, and the residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0% - 10% petroleum ether to afford rac-7-(fluoromethyl)-7-(nitromethyl)-1,4-dioxaspiro[4.5]decane (950 mg) as a colorless oil. LCMS (ESI-MS) m / z = 234.2 [M+H] + .

[0282] rac-(7-(Fluoromethyl)-1,4-dioxaspiro[4.5]decane-7-yl)methanamine In ethanol (10 mL) and water (3 mL), a solution of rac-7-(fluoromethyl)-7-(nitromethyl)-1,4-dioxaspiro[4.5]decane (950 mg, 4.6 mmol), iron (2.56 g, 46.7 mmol), and ammonium chloride (990 mg, 18.7 mmol) was stirred at 80 °C for 2 hours. The resulting mixture was filtered, and the filter cake was washed with ethanol (3 x 50 mL). The filtrate was concentrated under reduced pressure to afford rac-(7-(fluoromethyl)-1,4-dioxaspiro[4.5]decane-7-yl)methanamine (crude, 950 mg) as a yellow oil. LCMS (ESI-MS) m / z = 204.2 [M+H] + .

[0283] rac-3-(((7-(Fluoromethyl)-1,4-dioxaspiro[4.5]decane-7-yl)methyl)amino)-4-nitrobenzonitrile In acetonitrile (15 mL), a solution of rac-(7-(fluoromethyl)-1,4-dioxaspiro[4.5]decan-7-yl)methanamine (950 mg, 4.7 mmol), 3-fluoro-4-nitrobenzonitrile (776 mg, 4.7 mmol), and potassium carbonate (1.29 g, 9.3 mmol) was stirred at 40 °C overnight. The resulting mixture was concentrated under vacuum. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0% - 50% petroleum ether to afford rac-3-(((7-(fluoromethyl)-1,4-dioxaspiro[4.5]decan-7-yl)methyl)amino)-4-nitrobenzonitrile (1.0 g) as a yellow solid. LCMS (ESI-MS) m / z = 350.3 [M+H] + .

[0284] rac-4-Amino-3-(((7-(fluoromethyl)-1,4-dioxaspiro[4.5]decan-7-yl)methyl)amino)benzonitrile In ethanol (12 mL) and water (4 mL), a solution of rac-3-(((7-(fluoromethyl)-1,4-dioxaspiro[4.5]decan-7-yl)methyl)amino)-4-nitrobenzonitrile (1.0 g, 2.9 mmol), iron (1.60 g, 28.6 mmol), and ammonium chloride (0.61 g, 11.4 mmol) was stirred at 80 °C for 2 h. The resulting mixture was concentrated under vacuum to give the crude product rac-4-amino-3-(((7-(fluoromethyl)-1,4-dioxaspiro[4.5]decan-7-yl)methyl)amino)benzonitrile (1.0 g) as a yellow oil. LCMS (ESI-MS) m / z = 320.3 [M+H] + .

[0285] rac-1-((7-(fluoromethyl)-1,4-dioxaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile In 53165 trimethoxymethane (3 mL), a solution of rac-4-amino-3-(((7-(fluoromethyl)-1,4-dioxaspiro[4.5]decane-7-yl)methyl)amino)benzonitrile (1.0 g, 3.1 mmol) and formic acid (143 mg, 3.1 mmol) was stirred at 80 °C overnight. The resulting mixture was concentrated under vacuum. The residue was purified by preparative TLC (ethyl acetate) to give rac-1-((7-(fluoromethyl)-1,4-dioxaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (900 mg) as a colorless oil. LCMS (ESI-MS) m / z = 330.3 [M+H] + .

[0286] rac-1-((1-(fluoromethyl)-3-oxocyclohexyl)methyl)-1H-benzo[d]imidazole-6-carbonitrile In 53165 formic acid (5 mL), a solution of rac-1-((7-(fluoromethyl)-1,4-dioxaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (900 mg, 2.7 mmol) was stirred at room temperature overnight. The resulting mixture was concentrated under vacuum. The residue was purified by preparative TLC (100% ethyl acetate) to give rac-1-((1-(fluoromethyl)-3-oxocyclohexyl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (500 mg) as a white solid. LCMS (ESI-MS) m / z = 286.3 [M+H] + .

[0287] rac-1-(((3S,5R)-5-(fluoromethyl)-1-oxaspiro[2.5]octan-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile and rac-1-(((3R,5R)-5-(fluoromethyl)-1-oxaspiro[2.5]octan-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile In dimethyl sulfoxide (5 mL), a solution of rac-1-((1-(fluoromethyl)-3-oxocyclohexyl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (500 mg, 1.7 mmol) and iodomethyl methyl sulfoxide (424 mg, 1.9 mmol) was stirred at room temperature for 5 minutes. The solution was cooled to 0 °C and potassium tert-butoxide (216 mg, 1.9 mmol) was added dropwise over 2 minutes. The resulting mixture was stirred at 0 °C for an additional 1 hour. The reaction was quenched with water at 0 °C. The aqueous phase was extracted with ethyl acetate (3 × 100 mL). The resulting mixture was concentrated under vacuum. The residue was purified by preparative TLC (ethyl acetate = 100%) to give rac-1-(((3S,5R)-5-(fluoromethyl)-1-oxaspiro[2.5]octan-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (200 mg) as a white solid and rac-1-(((3R,5R)-5-(fluoromethyl)-1-oxaspiro[2.5]octan-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (180 mg) as a white solid. LCMS (ESI-MS) m / z = 300.3 [M+H] + .

[0288] rac-1-(((5S,7R)-7-(fluoromethyl)-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile In N-methyl-2-pyrrolidone (3 mL), a solution of ethyl carbamate (1.1 g, 13.3 mmol) and tert-butyllithium (107 mg, 1.3 mmol) was stirred at room temperature for 5 minutes. rac-1-(((3S,5R)-5-(fluoromethyl)-1-oxaspiro[2.5]octan-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (200 mg, 0.7 mmol) was added dropwise over 10 minutes. The resulting mixture was stirred at 75 °C overnight. The resulting mixture was cooled to room temperature and concentrated under vacuum to afford the crude product. The residue was purified by preparative TLC (dichloromethane:methanol = 10:1) to give rac-1-(((5S,7R)-7-(fluoromethyl)-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (200 mg) as a white solid. LCMS (ESI-MS) m / z = 343.3 [M+H] + .

[0289] rac-1-(((5S,7R)-7-(fluoromethyl)-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile In dioxane (3 mL), a solution of rac-1-(((5S,7R)-7-(fluoromethyl)-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (200 mg, 0.6 mmol), 2-(5-chloropyrazin-2-yl)propan-2-ol (151 mg, 0.9 mmol), N1,N2-dimethylethane-1,2-diamine (103 mg, 1.1 mmol), copper(I) iodide (111 mg, 0.6 mmol), and tripotassium phosphate (248 mg, 1.2 mmol) was stirred at 100 °C overnight under a nitrogen atmosphere. The resulting mixture was concentrated under vacuum. The residue was purified by preparative TLC (dichloromethane:methanol = 10:1) to give rac-1-(((5S,7R)-7-(fluoromethyl)-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (180 mg) as a white solid. LCMS (ESI-MS) m / z = 479.2 [M+H] + .

[0290] 1-(((5S,7R)-7-(fluoromethyl)-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile and 1-(((5R,7S)-7-(fluoromethyl)-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile A mixture of TIFF2025522953000143.tif68165rac-1-(((5S,7R)-7-(fluoromethyl)-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (180 mg, 0.4 mmol) was separated by preparative chiral HPLC under the following conditions: column: CHIRALPAK IE, 2 × 25 cm, 5 μm, mobile phase A: MtBE (0.5% 2M NH3-MeOH)--HPLC, mobile phase B: EtOH--HPLC, flow rate: 20 mL / min, gradient: 15% B to 15% B in 19 minutes, wavelength: 220 / 254 nm, RT1 (min): 12.796; RT2 (min): 15.536, sample solvent: MtBE (0.5% 2M NH3-MeOH)--HPLC, injection volume: 0.4 mL. The desired fractions were combined and lyophilized to obtain two products.

[0291] The isomer eluting first: 1-(((5R,7S)-7-(fluoromethyl)-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile as a white solid (62.2 mg, purity 98.2%, 100% ee). 1 1H NMR (400 MHz, dimethyl sulfoxide-d6) δ: 9.25 (s, 1H), 8.61 (s, 1H), 8.46 (s, 1H), 8.24 (s, 1H), 7.85 (d, J = 8.0 Hz, 1H), 7.61 (d, J = 8.4 Hz, 1H), 5.42 (s, 1H), 4.55 - 4.46 (m, 2H), 4.18 - 3.89 (m, 4H), 2.12 - 1.76 (m, 6H), 1.46 (s, 8H). LCMS (ESI-MS) m / z = 479.2 [M + H] + .

[0292] Second eluting isomer: 1-(((5S,7R)-7-(fluoromethyl)-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile as a white solid (63.0 mg, purity 90.9%, 100% ee). NMR (400 MHz, dimethyl sulfoxide-d6) δ: 9.25 (s, 1H), 8.62 (s, 1H), 8.47 (s, 1H), 8.24 (s, 1H), 7.85 (d, J = 8.4 Hz, 1H), 7.61 (d, J = 8.4 Hz, 1H), 5.44 (s, 1H), 4.56 - 4.46 (m, 2H), 4.19 (s, 1H), 4.07 - 4.03 (m, 2H), 3.90 (d J = 10.0 Hz, 1H), 2.12 - 2.03 (m, 1H), 2.02 - 1.96 (m, 1H), 1.95 - 1.89 (m, 1H), 1.89 - 1.76 (m, 2H), 1.46 (s, 9H)). LCMS (ESI-MS) m / z = 479.2 [M+H] + .

[0293] Example 8. Preparation of 1-(((1R,3S,6S)-3'-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-2'-oxospiro[bicyclo[4.1.0]heptane-3,5'-oxazolidine]-1-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000144.tif53165

[0294] Detailed procedure 3-methoxycyclohex-1,4-diene-1-carboxylic acid TIFF2025522953000145.tif37165Put m-anisic acid (50.0 g, 0.33 mol) and H2O (75 mL) into a three-necked flask. The flask is equipped with a mechanical stirrer, an ammonia inlet, and a cold finger condenser containing solid carbon dioxide and acetone. Pass ammonia over the solution through a glass filter and collect about 1 liter of ammonia. Replace the ammonia inlet with a nitrogen inlet and introduce a slow stream of nitrogen onto the surface. Start stirring vigorously (note: exothermic reaction) and add lithium wire (7.0 g, 1.0 mol) little by little over 20 - 30 minutes to perform Birch reduction. Continue vigorous stirring until the blue color completely disappears. Stop stirring, remove the condenser, and completely evaporate the ammonia under a constant stream. Then add 1 M potassium hydroxide solution (500 mL) (thoroughly purged with nitrogen) and start stirring again. Heat the solution in a water bath maintained at 60 °C for 2.5 hours. Replace the water bath with an ice bath and cool the solution to 10 °C. While continuing the ice bath, add concentrated HCl (100 mL) without allowing the temperature to exceed 70 °C. Continue stirring until the solution temperature drops to about 15 °C. Remove the ice bath and continue stirring until a homogeneous solution is obtained. Transfer the aqueous solution to a separatory funnel and extract with CH2Cl2 (3 x 200 mL). Combine the extracts, dry them, and evaporate them on a rotary evaporator under reduced pressure. Recrystallize a sample of the product from toluene. LCMS (ESI-MS) m / z = 155.2 [M+H] + .

[0295] 5-Oxocyclohex-1-ene-1-carboxylic acid In 37165 methanol (200 mL), oxalic acid (1.27 g, 14.1 mmol) and water (50 mL) were added to a solution of 3-methoxycyclohex-1,4-diene-1-carboxylic acid (14.5 g, 94.1 mmol). The resulting solution was stirred at room temperature for 16 h. The residue was concentrated and purified by silica gel column (EA:PE = 2:1) to give 5-oxocyclohex-1-ene-1-carboxylic acid (3 g) as a white solid. LCMS (ESI-MS) m / z = 141.0 [M+H] + .

[0296] Methyl 5-oxocyclohex-1-ene-1-carboxylate In 32165 diethyl ether (300 mL), diazomethane (27.0 g, 642 mmol) was added to a solution of 5-oxocyclohex-1-ene-1-carboxylic acid (30 g, 214 mmol). The resulting solution was stirred at room temperature for 16 h. The residue was concentrated and purified by silica gel column (EA:PE = 1:1) to give the desired product, methyl 5-oxocyclohex-1-ene-1-carboxylate (11.0 g) as a pale yellow oil. LCMS (ESI-MS) m / z = 155.1 [M+H] + .

[0297] Methyl 1,4-dioxaspiro[4.5]dec-7-ene-7-carboxylate In methylene chloride (50 mL), to a solution of methyl 5 - oxocyclohex - 1 - en - 1 - carboxylate (2.3 g, 14.9 mmol) were added ethylene glycol (1.39 g, 22.4 mmol), trimethyl orthoformate (2.37 g, 22.4 mmol) and methanesulfonic acid (220 mg, 2.24 mmol). The resulting solution was stirred at room temperature for 16 h. The residue was evaporated and purified by silica gel column (EA:PE = 1:1) to give methyl 1,4 - dioxaspiro[4.5]dec - 7 - ene - 7 - carboxylate (2.30 g) as a pale yellow oil. LCMS (ESI - MS) m / z = 199.0 [M + H] + .

[0298] (1,4 - Dioxaspiro[4.5]dec - 7 - en - 7 - yl)methanol In diethyl ether (20 mL), to a solution of methyl 1,4 - dioxaspiro[4.5]dec - 7 - ene - 7 - carboxylate (2.10 g, 10.6 mmol) was added lithium aluminum hydride (302 mg, 7.95 mmol). The resulting solution was stirred at room temperature for 2 h. The reaction was quenched with KOH (15%, 1 mL) and water (1 mL). The residue was extracted with Et2O (20.0 mL×2). The combined organic phases were evaporated and purified by silica gel column (EA:PE = 1:1) to give (1,4 - dioxaspiro[4.5]dec - 7 - en - 7 - yl)methanol (1.6 g) as a pale yellow oil.

[0299] rac - Spiro[bicyclo[4.1.0]heptane - 3,2’ - [1,3]dioxolane] - 1 - ylmethanol In methylene chloride (30 mL), to a solution of 1,4-dioxaspiro[4.5]decan-7-ylmethanol (1.60 g, 9.40 mmol) were added diethylzinc (2.90 g, 23.5 mmol), diiodomethane (6.29 g, 23.5 mmol) and trifluoroacetic acid (2.68 g, 23.5 mmol) at 0 °C. The resulting solution was stirred at room temperature for 16 h. The residue was evaporated and purified by silica gel column (EA:PE = 1:1) to give rac-spiro[bicyclo[4.1.0]heptane-3,2’-[1,3]dioxolan]-1-ylmethanol (1.00 g) as a pale yellow oil.

[0300] rac-2-(Spiro[bicyclo[4.1.0]heptane-3,2’-[1,3]dioxolan]-1-ylmethyl)isoindoline-1,3-dione In tetrahydrofuran (20 mL), to a solution of rac-spiro[bicyclo[4.1.0]heptane-3,2’-[1,3]dioxolan]-1-ylmethanol (1.0 g, 5.43 mmol) were added phthalimide (1.2 g, 8.16 mmol), diisopropyl azodicarboxylate (1.65 g, 8.14 mmol) and triphenylphosphine (2.14 g, 8.14 mmol). The resulting solution was stirred at 60 °C for 5 h. The solution was cooled to room temperature, the residue was concentrated and purified by silica gel column (EA:PE = 1:1) to give rac-2-(spiro[bicyclo[4.1.0]heptane-3,2’-[1,3]dioxolan]-1-ylmethyl)isoindoline-1,3-dione (1.40 g, 60.8%) as a white solid. LCMS (ESI-MS) m / z = 314.1 [M+H] + .

[0301] rac-spiro[bicyclo[4.1.0]heptane-3,2’-[1,3]dioxolan]-1-ylmethanamine In 42165 ethyl alcohol (20 mL), hydrazinium hydroxide (830 mg, 16.6 mmol) was added to a solution of rac-2-(spiro[bicylco[4.1.0]heptane-3,2’-[1,3]dioxolan]-1-ylmethyl)isoindole-1,3-dione (1.30 g, 4.15 mmol). The resulting solution was stirred at reflux for 5 hours. The mixture was cooled to room temperature, concentrated, and purified by silica gel column (DCM:MeOH = 10:1) to give rac-spiro[bicylco[4.1.0]heptane-3,2’-[1,3]-1-ylmethanamine (450 mg) as a pale yellow oil.

[0302] rac-4-nitro-3-((spiro[bicylco[4.1.0]heptane-3,2’-[1,3]dioxolan]-1-ylmethyl)amino)benzonitrile In acetonitrile (5 mL), 3-fluoro-4-nitrobenzonitrile (363 mg, 2.18 mmol) and potassium carbonate (649 mg, 6.55 mmol) were added to a solution of rac-spiro[bicylco[4.1.0]heptane-3,2’-[1,3]dioxolan]-1-ylmethanamine (400 mg, 2.18 mmol). The resulting solution was stirred at 40 °C for 4 hours. The mixture was cooled to room temperature, concentrated, and purified by silica gel column (EA:PE = 1:1) to give rac-4-nitro-3-((spiro[bicylco[4.1.0]heptane-3,2’-[1,3]dioxolan]-1-ylmethyl)amino)benzonitrile (650 mg) as a yellow solid. LCMS (ESI-MS) m / z = 330.1 [M+H] + .

[0303] rac-4-amino-3-((spiro[bicylco[4.1.0]heptane-3,2’-[1,3]dioxolan]-1-ylmethyl)amino)benzonitrile In ethyl alcohol (10 mL) and water (3 mL), iron (1.02 g, 18.2 mmol) and ammonium chloride (390 mg, 7.29 mmol) were added to a solution of rac-4-nitro-3-((spiro[bicyclo[4.1.0]heptane-3,2’-[1,3]dioxolan]-1-ylmethyl)amino)benzonitrile (600 mg, 1.82 mmol). The resulting solution was stirred at 80 °C for 4 h. The mixture was cooled to room temperature, concentrated, and purified by silica gel column (DCM:MeOH = 10:1) to give rac-4-amino-3-((spiro[bicyclo[4.1.0]heptane-3,2’-[1,3]dioxolan]-1-ylmethyl)amino)benzonitrile (500 mg) as a yellow oil. LCMS (ESI-MS) m / z = 300.0 [M+H] + .

[0304] rac-1-(Spiro[bicyclo[4.1.0]heptane-3,2’-[1,3]dioxolan]-1-ylmethyl)-1H-benzo[d]imidazole-6-carbonitrile In trimethyl orthoformate (5 mL), formic acid (69.2 mg, 1.5 mmol) was added to a solution of rac-4-amino-3-((spiro[bicyclo[4.1.0]heptane-3,2’-[1,3]dioxolan]-1-ylmethyl)amino)benzonitrile (450 mg, 1.5 mmol). The resulting solution was stirred at 80 °C for 16 h. The mixture was cooled to room temperature, concentrated, and purified by silica gel column (EA:PE = 1:1) to give rac-1-(spiro[bicyclo[4.1.0]heptane-3,2’-[1,3]dioxolan]-1-ylmethyl)-1H-benzo[d]imidazole-6-carbonitrile (350 mg) as a yellow solid. LCMS (ESI-MS) m / z = 310.1 [M+H] + .

[0305] rac-1-((3-oxobicyclo[4.1.0]heptan-1-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000156.tif53165 rac-1-(spiro[bicyclo[4.1.0]heptane-3,2’-[1,3]dioxolan]-1-ylmethyl)-1H-benzo[d]imidazole-6-carbonitrile (350 mg, 1.13 mmol) and formic acid (5.0 mL, 0.04 mmol) were combined in a 25 mL round-bottom flask. The resulting solution was stirred at room temperature for 2 hours. The reaction was quenched with saturated NaHCO3 and extracted with EA (20 mL X 2). The organic phases were combined, dried over anhydrous Na2SO4, and evaporated to give rac-1-((3-oxobicyclo[4.1.0]heptan-1-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (300 mg) as a pale yellow solid. LCMS (ESI-MS) m / z = 266.2 [M+H] + .

[0306] rac-1-(((1R,3S,6S)-spiro[bicyclo[4.1.0]heptane-3,2’-oxirane]-1-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile and rac-1-(((1R,3R,6S)-spiro[bicyclo[4.1.0]heptane-3,2’-oxirane]-1-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile In 53165 DMSO (5 mL), to a solution of trimethylsulfoxonium iodide (3.77 mmol) was added iodotrimethyl-λ6-sulfane (3.77 mmol) at 0 °C. After stirring for 30 minutes, rac-1-((3-oxobicyclo[4.1.0]heptan-1-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (3.77 mmol) was added. The resulting solution was stirred at room temperature for 1 hour and then quenched with water. The resulting mixture was extracted with EtOAc (3 × 5.0 mL). The combined organic layers were washed with brine (3 × 5.0 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure to obtain a mixture of rac-1-(((1R,3S,6S)-spiro[bicyclo][4.1.0]heptane-3,2’-oxirane]-1-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile and rac-1-(((1R,3R,6S)-spiro[bicyclo[4.1.0]heptane-3,2’-oxirane]-1-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (210 mg) as a pale yellow solid. LCMS (ESI-MS) m / z = 280.1 [M+H] + .

[0307] rac-1-(((1R,3S,6S)-2'-oxospiro[bicyclo[4.1.0]heptane-3,5'-oxazolidine]-1-yl)methyl)-1H-benzo[d]imidazole- 6-carbonitrile and rac-1-(((1R,3R,6S)-2'-oxospiro[bicyclo[4.1.0]heptane-3,5'-oxazolidine]-1-yl)methyl)-1H-benzo[ d]imidazole-6-carbonitrile In 58165 methylpyrrolidone (5 mL), to a solution of a mixture of rac-1-(((1R,3S,6S)-spiro[bicyclo[4.1.0]heptane-3,2’-oxirane]-1-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile and rac-1-(((1R,3R,6S)-spiro[bicyclo[4.1.0]heptane-3,2’-oxirane]-1-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (200 mg, 0.716 mmol) was added urethane (319 mg, 3.58 mmol) and tert-butoxylithium (143 mg, 1.79 mmol). The resulting solution was stirred at 100 °C for 16 h. The resulting mixture was cooled to room temperature, diluted with EtOAc (10 mL), washed with brine (3×10 mL), dried over anhydrous Na2SO4, concentrated, and purified by silica gel column (DCM:MeOH = 10:1) to give a mixture of rac-1-(((1R,3S,6S)-2’-oxospiro[bicyclo[4.1.0]heptane-3,5’-oxazolidine]-1-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile and rac-1-(((1R,3R,6S)-2’-oxospiro[bicyclo[4.1.0]heptane-3,5’-oxazolidine]-1-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (200 mg) as a pale yellow solid. LCMS (ESI-MS) m / z = 323.1 [M+H] + .

[0308] rac-1-(((1R,3S,6S)-3’-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-2’-oxospiro[bicyclo[4.1.0]heptane-3,5’-oxazolidine]-1-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile and rac-1-(((1R,3R,6S)-3’-(5-(2-hydroxypropane)-2-yl)pyrazin-2-yl)-2’-oxospiro[bicyclo[4.1.0]heptane-3,5’-oxazolidine]-1-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile In TIFF2025522953000159.tif58165, to a solution of rac-1-(((1R,3S,6S)-2'-oxospiro[bicyclo[4.1.0]heptane-3,5'-oxazolidin]-1-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile and rac-1-(((1R,3R,6S)-2'-oxospiro[bicyclo[4.1.0]heptane-3,5'-oxazolidin]-1-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (200 mg, 0.62 mmol) in dioxane (5.0 mL), 2-(5-bromopyrazin-2-yl)propan-2-ol (135 mg, 0.62 mmol), N1,N2-dimethylethane-1,2-diamine (109 mg, 1.24 mmol), tripotassium phosphate (263 mg, 1.24 mmol), and copper(I) iodide (118 mg, 0.62 mmol) were added. The resulting solution was stirred at 100 °C for 16 h. The residue was evaporated and purified by silica gel column (DCM:MeOH = 10:1) to give rac-1-(((1R,3S,6S)-3'-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-2'-oxospiro[bicyclo[4.1.0]heptane-3,5'-oxazolidin]-1-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile and rac-1-(((1R,3R,6S)-3'-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-2'-oxospiro[bicyclo[4.1.0]heptane-3,5'-oxazolidin]-1-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (100 mg) as a pale yellow solid. LCMS (ESI-MS) m / z = 459.2 [M+H] + .

[0309] A chiral separation: A mixture of rac-1-(((1R,3S,6S)-3’-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-2’-oxospiro[bicyclo[4.1.0]heptane-3,5’-oxazolidin]-1-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile and rac-1-(((1R,3R,6S)-3’-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-2’-oxospiro[bicyclo[4.1.0]heptane-3,5’-oxazolidin]-1-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (100 mg, 0.22 mmol) was separated by preparative chiral HPLC under the conditions: column: CHIRALPAK IF-3, 4.6 * 50 mm, 3 μm, mobile phase A: MTBE (0.1% DEA): ethanol = 70:30, flow rate: 1 mL / min, gradient: 0%B to 0%B; injection volume: 5 μl to obtain the first eluting isomer: rac-1-(((1R,3S,6S)-3’-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-2’-oxospiro[bicyclo[4.1.0]heptane-3,5’-oxazolidin]-1-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (10 mg) as a white solid, and the second eluting isomer: rac-1-(((1R,3R,6S)-3’-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-2’-oxospiro[bicyclo[4.1.0]heptane-3,5’-oxazolidin]-1-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (15 mg) as a white solid.

[0310] 1-(((1R,3S,6S)-3'-(5-(2-Hydroxypropan-2-yl)pyrazin-2-yl)-2'-oxospiro[bicyclo[4.1.0]heptane-3,5'-oxazolidine]-1-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile and 1-(((1S,3R,6R)-3'-(5-(2-Hydroxypropan-2-yl)pyrazin-2-yl)-2'-oxospiro[bicyclo[4.1.0]heptane-3,5'-oxazolidine]-1-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000160.tif63165 rac-1-(((1R,3S,6S)-3'-(5-(2-Hydroxypropan-2-yl)pyrazin-2-yl)-2'-oxospiro[bicyclo[4.1.0]heptane-3,5'-oxazolidine]-1-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (10.0 mg, 0.0220 mmol) was separated by preparative chiral HPLC under the following conditions: column: CHIRALPAK IF-3 4.6×50 mm, 3 um, mobile phase A: MTBE (0.1% DEA): ethanol = 70:30, flow rate: 1 mL / min, injection volume: 5 μL. The desired fractions were combined and lyophilized to give two products.

[0311] The first eluting isomer: 1-(((1S,3R,6R)-3'-(5-(2-Hydroxypropan-2-yl)pyrazin-2-yl)-2'-oxospiro[bicyclo(bicyclo) [4.1.0]heptane-3,5'-oxazolidine]-1-yl)methyl)-1H-bbenzo[d]imidazole-6-carbonitrile (4.10 mg) as a white solid. 11H NMR (400 MHz, DMSO-d6) δ 8.95 (s, 1H), 8.61 (s, 1H), 8.48 (s, 1H), 8.35 (s, 1H), 7.65 (d, J = 8.4 Hz, 1H), 7.32 (d, J = 10.0 Hz, 1H), 5.44 (s, 1H), 4.37-4.33 (m, 1H), 4.09-4.05 (m, 1H), 3.66-3.64 (m, 1H), 2.93-2.91 (m, 1H), 2.18-2.14 (m, 1H), 2.01-1.87 (m, 3H), 1.65-1.56 (m, 2H), 1.52 (s, 3H), 1.45 (s, 3H),1.43-1.37 (m, 1H), 0.97-0.94 (m, 1H), 0.68-0.66 (m, 1H). LCMS (ESI-MS) m / z = 459.2 [M+H] + .

[0312] Second eluting isomer: 1-(((1R,3S,6S)-3'-(5-(2-Hydroxypropan-2-yl)pyrazin-2-yl)-2'-oxospiro[bicyclo[4.1.0]heptane-3,5'-oxazolidin]-1-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (3.60 mg) as a white solid. 11H NMR (400 MHz, DMSO-d6) δ 8.95 (s, 1H), 8.61 (s, 1H), 8.48 (s, 1H), 8.35 (s, 1H), 7.65 (d, J = 8.4 Hz, 1H), 7.32 (d, J = 10.0 Hz, 1H), 5.44 (s, 1H), 4.37-4.33 (m, 1H), 4.09-4.05 (m, 1H), 3.66-3.64 (m, 1H), 2.93-2.91 (m, 1H), 2.18-2.14 (m, 1H), 1.91-1.87 (m, 3H), 1.61-1.58 (m, 2H), 1.52 (s, 3H), 1.45 (s, 3H), 1.43-1.39 (m, 1H), 0.97-0.95 (m, 1H), 0.71-0.65 (m, 1H). LCMS (ESI-MS) m / z = 459.3 [M+H] + .

[0313] Example 9. Preparation of 1-(((5S,7R)-3-(5-(2-Hydroxypropan-2-yl)pyrazin-2-yl)-7-methoxy-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000161.tif53165

[0314] Detailed procedure 1,4-Dioxaspiro[4.5]decane-7-one TIFF2025522953000162.tif 32165 p - Toluenesulfonic acid (13.8 g, 80.3 mmol) was added to a mixture of cyclohexane - 1,3 - dione (30 g, 268 mmol), molecular sieves (4 Å) (30 g), and magnesium sulfate (30 g) in ethylene glycol (300 mL). The resulting mixture was stirred at room temperature overnight. The reaction mixture was diluted with water (300 mL), and the resulting mixture was extracted with ethyl acetate (3 x 300 mL). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the crude product 1,4 - dioxaspiro[4.5]decan - 7 - one (crude, 28 g) as a pale yellow oil. LCMS (ESI - MS) m / z = 157.1 [M + H] + .

[0315] rac - 1,6,9 - Trioxadispiro[2.1.4 5 .3 3 dodecane TIFF2025522953000163.tif 37165 Potassium tert - butoxide (22.1 g, 197 mmol) was added to a solution of 1,4 - dioxaspiro[4.5]decan - 7 - one (28 g, 179 mmol) and trimethylsulfoxonium iodide (39.5 g, 179 mmol) in dimethyl sulfoxide (280 mL). The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was diluted with water (300 mL), and the resulting solution was extracted with ethyl acetate (3 x 300 mL). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the crude product rac - 1,6,9 - trioxadispiro[2.1.4 5 .3 3 dodecane (crude, 28 g) as a pale yellow oil. LCMS (ESI - MS) m / z = 171.1 [M + H] + .

[0316] rac - 7 - (Aminomethyl)-1,4 - dioxaspiro[4.5]decan - 7 - ol TIFF2025522953000164.tif37165A solution of rac-1,6,9-trioxadispiro[2.1.4 5 .3 3 dodecane (28 g, 165 mmol) in ammonia (7N, 280 mL, 1.96 mol) in methanol. The resulting mixture was heated to 80 °C and stirred overnight. The reaction mixture was concentrated under vacuum to give the crude product rac-7-(aminomethyl)-1,4-dioxaspiro[4.5]decan-7-ol (crude, 28 g) as a pale yellow oil. LCMS (ESI-MS) m / z = 188.1 [M+H] + .

[0317] rac-3-(((7-Hydroxy-1,4-dioxaspiro[4.5]decan-7-yl)methyl)amino)-4-nitrobenzonitrile TIFF2025522953000165.tif48165Potassium carbonate (41.3 g, 299 mmol) was added to a solution of rac-7-(aminomethyl)-1,4-dioxaspiro[4.5]decan-7-ol (28 g, 150 mmol) and 3-fluoro-4-nitrobenzonitrile (24.8 g, 150 mmol) in acetonitrile (200 mL). The resulting mixture was heated to 40 °C and stirred overnight. After cooling to room temperature, the resulting mixture was purified by silica gel column chromatography, eluting with ethyl acetate in 0% - 15% petroleum ether to give the desired product rac-3-(((7-hydroxy-1,4-dioxaspiro[4.5]decan-7-yl)methyl)amino)-4-nitrobenzonitrile (12 g) as an orange solid. LCMS (ESI-MS) m / z = 334.1 [M+H] + .

[0318] rac-4-Amino-3-(((7-hydroxy-1,4-dioxaspiro[4.5]decan-7-yl)methyl)amino)benzonitrile TIFF2025522953000166.tif53165 Iron powder (20.1 g, 360 mmol) was added to a solution of rac-3-(((7-hydroxy-1,4-dioxaspiro[4.5]decan-7-yl)methyl)amino)-4-nitrobenzonitrile (12 g, 36.0 mmol), ammonium chloride (7.7 g, 144 mmol), ethanol (120 mL), and water (40 mL). The resulting mixture was heated to 80 °C and stirred for 2 h. After cooling to room temperature, the resulting mixture was filtered and the filter cake was washed with ethanol (2 x 200 mL). The filtrate was concentrated under reduced pressure to afford the crude product rac-4-amino-3-(((7-hydroxy-1,4-dioxaspiro[4.5]decan-7-yl)methyl)amino)benzonitrile (crude, 12 g) as a yellow solid. LCMS (ESI-MS) m / z = 304.2 [M+H] + .

[0319] rac-1-((7-hydroxy-1,4-dioxaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000167.tif53165 Formic acid (1.82 g, 39.6 mmol) was added to a solution of rac-4-amino-3-(((7-hydroxy-1,4-dioxaspiro[4.5]decan-7-yl)methyl)amino)benzonitrile (12 g, 39.6 mmol) in trimethoxymethane (83.9 g, 791 mmol). The resulting mixture was heated to 80 °C and stirred for 3 h. The mixture was cooled to room temperature and concentrated to afford a crude product. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0% - 70% petroleum ether, to afford the desired product rac-1-((7-hydroxy-1,4-dioxaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (9 g) as a white solid. LCMS (ESI-MS) m / z = 314.1 [M+H] + .

[0320] rac-1-((1-Hydroxy-3-oxocyclohexyl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000168.tif53165 In acetone (30 mL) and water (10 mL), a solution of rac-1-((7-hydroxy-1,4-dioxaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (5 g, 17.7 mmol) was treated with 1 M aqueous hydrochloric acid (8.66 mL, 8.66 mmol), and the resulting mixture was stirred at room temperature overnight. The reaction mixture was diluted with water (100 mL) and then extracted with ethyl acetate (3 x 100 mL). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give the crude product. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0% - 80% petroleum ether to afford the desired product rac-1-((1-hydroxy-3-oxocyclohexyl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (3 g) as a yellow solid. LCMS (ESI-MS) m / z = 270.1 [M+H] + .

[0321] rac-1-(((3S,5R)-5-Hydroxy-1-oxaspiro[2.5]octane-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000169.tif53165 Potassium tert-butoxide (1.37 g, 12.3 mmol) was added to a solution of rac-1-((1-hydroxy-3-oxocyclohexyl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (3 g, 11.1 mmol) and trimethylsulfoxonium iodide (2.69 g, 12.3 mmol) in dimethyl sulfoxide (30 mL). The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was diluted with water (50 mL), and the resulting solution was extracted with ethyl acetate (3 x 50 mL). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give the crude product rac-1-(((3S,5R)-5-hydroxy-1-oxaspiro[2.5]octan-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (2 g) as a pale yellow oil. LCMS (ESI-MS) m / z = 284.1 [M+H] + .

[0322] rac-1-(((5S,7R)-7-hydroxy-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile Lithium tert-butoxide (1.13 g, 14.1 mmol) was added to a solution of ethyl carbamate (12.6 g, 141 mmol) in N-methylpyrrolidone (10 mL). After stirring for 5 minutes at room temperature, a solution of rac-1-(((3S,5R)-5-hydroxy-1-oxaspiro[2.5]octan-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (2 g, 7.06 mmol) in N-methylpyrrolidone (5 mL) was added dropwise. Subsequently, the reaction mixture was heated to 100 °C and stirred overnight. After cooling to room temperature, the residue was purified by reverse-phase flash chromatography under the following conditions: column, C18 silica gel; mobile phase, ACN in water (10 mmol / L NH4HCO3), gradient 0% - 30% in 10 minutes; detector, UV 254 nm to give the desired product rac-1-(((5S,7R)-7-hydroxy-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (1 g) as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 9.21 (d, J = 1.6 Hz, 1H), 8.60 (d, J = 1.6 Hz, 1H), 8.35 (d, J = 1.6 Hz, 1H), 8.29 (s, 1H), 6.45 (s, 1H), 4.50 (s, 1H), 3.80 (dd, J = 8.3, 8.3 Hz, 2H), 3.10 (dd, J = 8.1, 8.1 Hz, 2H), 2.10 - 2.00 (m, 2H), 1.60 - 1.48 (m, 2H), 1.45 - 1.43 (m, 4H). LCMS (ESI-MS) m / z = 327.1 [M+H] + .

[0323] rac-1-(((5S,7R)-7-hydroxy-3-(5-(2-((4-methoxybenzyl)oxy)propan-2-yl)pyrazin-2-yl)-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile Copper(I) iodide (582 mg, 3.06 mmol) was added to a mixture of rac-1-(((5S,7R)-7-hydroxy-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (1 g, 3.06 mmol), 2-chloro-5-(2-((4-methoxybenzyl)oxy)propan-2-yl)pyrazine (896 mg, 3.06 mmol), N 1 ,N 2 -dimethylethane-1,2-diamine (540 mg, 6.12 mmol) and tripotassium phosphate (1.3 g, 6.12 mmol) under a nitrogen atmosphere. The resulting mixture was heated to 100 °C and stirred overnight. After cooling to room temperature, the resulting mixture was filtered and concentrated under vacuum. The residue was purified by silica gel column chromatography, eluting with methanol in dichloromethane from 0% to 3% to give the desired product rac-1-(((5S,7R)-7-hydroxy-3-(5-(2-((4-methoxybenzyl)oxy)propan-2-yl)pyrazin-2-yl)-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (700 mg) as a pale yellow solid. 11H NMR (300 MHz, DMSO-d6) δ 9.21 (d, J = 1.6 Hz, 1H), 8.60 (d, J = 1.6 Hz, 1H), 8.35 (d, J = 1.6 Hz, 1H), 8.29 (s, 1H), 7.84 (d, J = 8.3 Hz, 1H), 7.59 (dd, J = 8.3, 1.5 Hz, 1H), 6.89 (d, J = 7.2 Hz, 2H), 6.79 (d, J = 8.3, 1.5 Hz, 1H), 7.2 Hz, 2H), 4.81 (s, 2H), 4.50 (s, 1H), 4.44 (d, J = 15.1 Hz, 1H), 4.36 (d, J = 15.0 Hz, 1H), 3.92 (d, J = 10.2 Hz, 1H), 3.84 (d, J = 10.2 Hz, 1H), 3.79 (s, 3H), 2.27 (d, J = 15.1 Hz, 1H), 2.00 (d, J = 13.4 Hz, 1H), 1.80 (d, J = 15.1 Hz, 1H), 1.69 (t, J = 12.9 Hz, 1H), 1.61 - 1.47 (m, 1H), 1.47 - 1.43 (m, 9H). LCMS (ESI-MS) m / z = 583.3 [M+H] + .

[0324] rac-1-(((5S,7R)-7-Methoxy-3-(5-(2-((4-methoxybenzyl)oxy)propan-2-yl)pyrazin-2-yl)-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000172.tif68165Iodomethane (170.33 mg, 1.20 mmol) was added to a mixture of rac-1-(((5S,7R)-7-hydroxy-3-(5-(2-((4-methoxybenzyl)oxy)propan-2-yl)pyrazin-2-yl)-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (700 mg, 1.20 mmol) and sodium hydride (60% in mineral oil, 48 mg, 1.20 mmol) in N,N-dimethylformamide (5 mL) at 0 °C. The mixture was stirred at 0 °C for 1 hour. The mixture was quenched slowly with water (10 mL) and extracted with ethyl acetate (3 x 10 mL). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give the crude product. The residue was purified by silica gel column chromatography, eluting with methanol in dichloromethane from 0% to 2%, to afford the desired product rac-1-(((5S,7R)-7-methoxy-3-(5-(2 -((4-methoxybenzyl)oxy)propan-2-yl)pyrazin-2-yl)-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (400 mg) as a white solid. 11H NMR (400 MHz, DMSO-d6) δ 9.21 (d, J = 1.6 Hz, 1H), 8.60 (d, J = 1.6 Hz, 1H), 8.35 (d, J = 1.6 Hz, 1H), 8.29 (s, 1H), 7.84 (d, J = 8.3 Hz, 1H), 7.59 (dd, J = 8.3, 1.5 Hz, 1H), 6.89 (d, J = 7.2 Hz, 2H), 6.79 (d, J = 8.3, 1.5 Hz, 1H) 7.2 Hz, 2H), 4.70 (s, 2H), 4.44 (d, J = 15.1 Hz, 1H), 4.36 (d, J = 15.0 Hz, 1H), 3.92 (d, J = 10.2 Hz, 1H), 3.84 (d, J = 10.2 Hz, 1H), 3.79 (s, 3H), 3.39 (s, 3H), 2.27 (d, J = 15.1 Hz, 1H), 2.00 (d, J = 13.4 Hz, 1H), 1.80 (d, J = 15.1 Hz, 1H), 1.69 (t, J = 12.9 Hz, 1H), 1.61 - 1.47 (m, 1H) 1.47 - 1.43 (m, 9H). LCMS (ESI-MS) m / z = 597.3 [M+H] + .

[0325] rac-1-(((5S,7R)-3-(5-(2-Hydroxypropan-2-yl)pyrazin-2-yl)-7-methoxy-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000173.tif68165Trifluoroacetic acid (229 mg, 2.01 mmol) was added to a solution of rac-1-(((5S,7R)-7-methoxy-3-(5-(2-((4-methoxybenzyl)oxy)propan-2-yl)pyrazin-2-yl)-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (400 mg, 0.67 mmol) in dichloromethane (3 mL). The resulting mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated under vacuum and purified by silica gel column chromatography, eluting with methanol in dichloromethane from 0% to 3% to give the desired product rac-1-(((5S,7R)-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methoxy-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (200 mg) as a yellow oil. 1 1H NMR (300 MHz, DMSO-d6) δ 9.21 (d, J = 1.6 Hz, 1H), 8.60 (d, J = 1.6 Hz, 1H), 8.35 (d, J = 1.6 Hz, 1H), 8.29 (s, 1H), 7.84 (d, J = 8.3 Hz, 1H), 7.59 (dd, J = 8.3, 1.5 Hz, 1H), 5.40 (s, 1H), 4.44 (d, J = 15.1 Hz, 1H), 4.36 (d, J = 15.0 Hz, 1H), 3.92 (d, J = 10.2 Hz, 1H), 3.84 (d, J = 10.2 Hz, 1H), 3.39 (s, 3H), 2.27 (d, J = 15.1 Hz, 1H), 2.00 (d, J = 13.4 Hz, 1H), 1.80 (d, J = 15.1 Hz, 1H), 1.69 (t, J = 12.9 Hz, 1H), 1.61 - 1.47 (m, 1H) 1.47 - 1.43 (m, 9H). LCMS (ESI-MS) m / z = 477.2 [M+H] + .

[0326] 1-(((5S,7R)-3-(5-(2-Hydroxypropan-2-yl)pyrazin-2-yl)-7-methoxy-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile and 1-(((5R,7S)-3-(5-(2-Hydroxypropan-2-yl)pyrazin-2-yl)-7-methoxy-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile A mixture of TIFF2025522953000174.tif58165rac-1-(((5S,7R)-3-(5-(2-Hydroxypropan-2-yl)pyrazin-2-yl)-7-methoxy-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (200 mg, 0.42 mmol) was separated by preparative chiral HPLC under the conditions: CHIRALPAK IE, 2 x 25 cm, 5 μm, mobile phase A: MTBE (0.5% 2M NH3-MeOH)--HPLC, mobile phase B: EtOH--HPLC, flow rate: 17 mL / min, gradient: 10% B to 10% B in 24 min, wavelength: 220 / 254 nm, RT1 (min): 14.661, RT2 (min): 20.017, sample solvent: EtOH--HPLC, injection volume: 0.5 mL. The desired fractions were combined and lyophilized to obtain the product.

[0327] The first eluting isomer: 1-(((5R,7S)-3-(5-(2-Hydroxypropan-2-yl)pyrazin-2-yl)-7-methoxy-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile as a white solid (44.6 mg, purity 99.7%, 100% ee). 11H NMR (300 MHz, DMSO-d6) δ 9.21 (d, J = 1.6 Hz, 1H), 8.60 (d, J = 1.6 Hz, 1H), 8.35 (d, J = 1.6 Hz, 1H), 8.29 (s, 1H), 7.84 (d, J = 8.3 Hz, 1H), 7.59 (dd, J = 8.3, 1.5 Hz, 1H), 5.40 (s, 1H), 4.44 (d, J = 15.1 Hz, 1H), 4.36 (d, J = 15.0 Hz, 1H), 3.92 (d, J = 10.2 Hz, 1H), 3.84 (d, J = 10.2 Hz, 1H), 3.39 (s, 3H), 2.27 (d, J = 15.1 Hz, 1H), 2.00 (d, J = 13.4 Hz, 1H), 1.80 (d, J = 15.1 Hz, 1H), 1.69 (t, J = 12.9 Hz, 1H), 1.61 - 1.47 (m, 1H), 1.47 - 1.43 (m, 9H). LCMS (ESI-MS) m / z = 477.2 [M+H] + .

[0328] Second eluting isomer: 1 - (((5S,7R)-3-(5-(2-hydroxypropan - 2 - yl)pyrazin - 2 - yl)-7 - methoxy - 2 - oxo - 1 - oxa - 3 - azaspiro[4.5]decane - 7 - yl)methyl)-1H - benzo[d]imidazole - 6 - carbonitrile as a white solid (41.0 mg, purity 98.6%, 100% ee). 11H NMR (300 MHz, DMSO-d6) δ 9.21 (d, J = 1.6 Hz, 1H), 8.60 (d, J = 1.6 Hz, 1H), 8.35 (d, J = 1.6 Hz, 1H), 8.29 (s, 1H), 7.84 (d, J = 8.3 Hz, 1H), 7.59 (dd, J = 8.3, 1.5 Hz, 1H), 5.40 (s, 1H), 4.44 (d, J = 15.1 Hz, 1H), 4.36 (d, J = 15.0 Hz, 1H), 3.92 (d, J = 10.2 Hz, 1H), 3.84 (d, J = 10.2 Hz, 1H), 3.39 (s, 3H), 2.27 (d, J = 15.1 Hz, 1H), 2.00 (d, J = 13.4 Hz, 1H), 1.80 (d, J = 15.1 Hz, 1H), 1.69 (t, J = 12.9 Hz, 1H), 1.61 - 1.47 (m, 1H), 1.47 - 1.43 (m, 9H). LCMS (ESI-MS) m / z = 477.2 [M+H] + .

[0329] Example 10. Preparation of 1 - (((5S,7S,8R)-3-(5-(2 - hydroxypropan - 2 - yl)pyrazin - 2 - yl)-8 - methoxy - 7 - methyl - 2 - oxo - 1 - oxa - 3 - azaspiro[4.5]decane - 7 - yl)methyl)-1H - benzo[d]imidazole - 6 - carbonitrile TIFF2025522953000175.tif53165

[0330] Detailed procedure rac - 1 - (((5S,7S,8R)-8 - hydroxy - 3-(5-(2 - ((4 - methoxybenzyl)oxy)propan - 2 - yl)pyrazin - 2 - yl)-7 - methyl - 2 - oxo - 1 - oxa - 3 - azaspiro[4.5]decane - 7 - yl)methyl)-1H - benzo[d]imidazole - 6 - carbonitrile Copper(I) iodide (448 mg, 2.35 mmol) was added under a nitrogen atmosphere to a solution of rac-1-(((5S,7S,8R)-8-hydroxy-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (800 mg, 2.35 mmol), 2-chloro-5-(2-((4-methoxybenzyl)oxy)propan-2-yl)pyrazine (686 mg, 2.35 mmol), N 1 ,N 2 -dimethylethane-1,2-diamine (414 mg, 4.70 mmol) and tripotassium phosphate (996 mg, 4.70 mmol) in 1,4-dioxane (10 mL). The resulting mixture was heated to 100 °C and stirred overnight. After cooling to room temperature, the resulting mixture was filtered and concentrated under vacuum. The residue was purified by silica gel column chromatography, eluting with methanol in 0% - 3% dichloromethane to give the desired product rac-1-(((5S,7S,8R)-8-hydroxy-3-(5-(2-((4-methoxybenzyl)oxy)propan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (1 g) as a pale yellow solid. LCMS (ESI-MS) m / z = 597.3 [M+H] + .

[0331] rac-1-(((5S,7S,8R)-8-hydroxy-3-(5-(2-((4-methoxybenzyl)oxy)propan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile and rac-1-(((5R,7S,8R)-8-hydroxy-3-(6-(2-((4-methoxybenzyl)oxy)propan-2-yl)pyridin-3-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile A mixture of TIFF2025522953000177.tif68165rac-1-(((5S,7S,8R)-8-hydroxy-3-(5-(2-((4-methoxybenzyl)oxy)propan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (1 g, 1.68 mmol) was separated by preparative chiral HPLC under the conditions: column: CHIRALPAK IE, 2x25 cm, 5 μm, mobile phase A: Hex: DCM = 3:1 (0.5% 2M NH3-MeOH)--HPLC, mobile phase B: IPA--HPLC, flow rate: 20 mL / min, gradient: 20% B to 20% B in 21 minutes, wavelength: 220 / 254 nm, RT1 (min): 10.051, RT2 (min): 16.651, sample solvent: EtOH--HPLC, injection volume: 1 mL. The desired fractions were combined and lyophilized to obtain the product.

[0332] The first eluting isomer: rac-1-(((5S,7S,8R)-8-hydroxy-3-(5-(2-((4-methoxybenzyl)oxy)propan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (200 mg) as a pale yellow solid. 11H NMR (400 MHz, DMSO-d6) δ 9.25 (d, J = 1.5 Hz, 1H), 8.56 (d, J = 1.5 Hz, 1H), 8.35 (d, J = 15.2 Hz, 1H), 8.23 (d, J = 1.6 Hz, 1H), 7.84 (d, J = 8.4 Hz, 1H), 7.59 (dd, J = 8.3, 1.5 Hz, 1H), 7.29 - 7.20 (m, 2H), 6.94 - 6.84 (m, 2H), 4.35 - 4.24 (m, 1H), 4.27 - 4.17 (m, 2H), 3.88 - 3.70 (m, 2H), 3.74 (s, 3H), 3.36 - 3.30 (m, 3H), 2.08 - 1.91 (m, 4H), 1.65 - 1.50 (m, 8H), 1.11 (s, 3H). LCMS (ESI-MS) m / z = 597.3 [M+H] + .

[0333] Second eluting isomer: rac-1-(((5R,7S,8R)-8-Hydroxy-3-(6-(2-((4-methoxybenzyl)oxy)propan-2-yl)pyridin-3-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile as a pale yellow solid (400 mg, yield 40.0%). 1 1H NMR (400 MHz, DMSO-d6) δ 9.30 (d, J = 1.5 Hz, 1H), 8.59 (d, J = 1.5 Hz, 1H), 8.42 (s, 1H), 8.25 (s, 1H), 7.83 (d, J = 8.1 Hz, 1H), 7.64 - 7.55 (m, 1H), 7.28 - 7.20 (m, 2H), 6.93 - 6.86 (m, 2H), 4.41 - 4.32 (m, 1H), 4.25 (s, 2H), 3.91 - 3.81 (m, 2H), 3.79 (s, 3H), 3.37 - 3.34 (m, 3H), 2.00 - 1.83 (m, 4H), 1.75 - 1.59 (m, 8H), 0.91 (s, 3H). LCMS (ESI-MS) m / z = 597.3 [M+H]+ .

[0334] rac-1-(((5S,7S,8R)-8-Methoxy-3-(5-(2-((4-methoxybenzyl)oxy)propan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000178.tif68165Iodomethane (35.5 mg, 0.25 mmol) was added to a solution of rac-1-(((5S,7S,8R)-8-hydroxy-3-(5-(2-((4-methoxybenzyl)oxy)propan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (150 mg, 0.25 mmol) and sodium hydride (60% in mineral oil, 10 mg, 0.25 mmol) in N,N-dimethylformamide (2 mL) at 0 °C. The resulting mixture was warmed to room temperature and stirred for 2 h. The mixture was quenched with 5 mL of water and the resulting solution was extracted with ethyl acetate (3 x 5 mL). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give the crude product. The residue was purified by silica gel column chromatography, eluting with methanol in dichloromethane from 0% to 2%, to give the desired product rac-1-(((5S,7S,8R)-8-methoxy-3-(5-(2-((4-methoxybenzyl)oxy)propan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (80 mg) as a white solid. 11H NMR (400 MHz, DMSO-d6) δ 9.25 (d, J = 1.5 Hz, 1H), 8.56 (d, J = 1.5 Hz, 1H), 8.35 (d, J = 15.2 Hz, 1H), 8.23 (d, J = 1.6 Hz, 1H), 7.84 (d, J = 8.4 Hz, 1H), 7.59 (dd, J = 8.3, 1.5 Hz, 1H), 7.29 - 7.20 (m, 2H), 6.94 - 6.84 (d, J = 8.3, 1.5 Hz, 1H), 6.94 - 6.84 (m, 2H), 4.30 - 4.17 (m, 4H), 3.88 - 3.70 (m, 2H), 3.74 (s, 3H), 3.39 (s, 3H), 3.10 - 3.03 (m, 1H), 2.08 - 1.91 (m, 3H, m, 4H), 1.65 - 1.50 (m, 8H), 1.11 (s, 3H). LCMS (ESI-MS) m / z = 611.3 [M+H] + .

[0335] rac-1-(((5S,7S,8R)-3-(5-(2-Hydroxypropan-2-yl)pyrazin-2-yl)-8-methoxy-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000179.tif68165Trifluoroacetic acid (44 mg, 0.39 mmol) was added to a solution of rac-1-(((5S,7S,8R)-8-methoxy-3-(5-(2-((4-methoxybenzyl)oxy)propan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (80 mg, 0.13 mmol) in dichloromethane (1 mL). The resulting mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated under vacuum to afford the crude product rac-1-(((5S,7S,8R)-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-8-methoxy-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (crude, 70 mg) as a yellow oil. LCMS (ESI-MS) m / z = 491.2 [M+H] + .

[0336] 1-(((5S,7S,8R)-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-8-methoxy-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile and 1-(((5R,7R,8S)-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-8-methoxy-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile A mixture of TIFF2025522953000180.tif68165rac-1-(((5S,7S,8R)-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-8-methoxy-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (70 mg, 0.14 mmol) was separated by preparative chiral HPLC under the conditions: column: CHIRAL ART Cellulose-SC, 2x25 cm, 5 μm, mobile phase A: Hex: DCM = 3:1 (0.5% 2M NH3-MeOH)--HPLC, mobile phase B: EtOH--HPLC, flow rate: 20 mL / min, gradient: 30%B - 30%B in 14 min, wavelength: 220 / 254 nm, RT1 (min): 9.92, RT2 (min): 12.16, sample solvent: EtOH--HPLC; injection volume: 0.8 mL. The desired fractions were combined and lyophilized to give the product.

[0337] The first eluting isomer: 1-(((5R,7R,8S)-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-8-methoxy-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile as a white solid (14.0 mg, purity 99.7%, 100% ee). 11H NMR (300 MHz, DMSO-d6) δ 9.18 (d, J = 1.5 Hz, 1H), 8.58 (d, J = 1.5 Hz, 1H), 8.37 (s, 1H), 8.25 - 8.20 (m, 1H), 7.84 (d, J = 8.3 Hz, 1H), 7.60 (dd, J = 8.3, 1.5 Hz, 1H), 5.42 (s, 1H), 4.31 (d, J = 14.5 Hz, 1H), 4.21 (dd, J = 8.3, 1.5 Hz, 1H), 5.42 (s, 1H), 4.21 (d, J = 14.5 Hz, 1H) d, J = 14.5 Hz, 1H), 3.81 (q, J = 10.3 Hz, 2H), 3.31 (s, 3H), 3.07 - 3.00 (m, 1H), 2.04 (d, J = 9.7 Hz, 2H), 1.95 (d, J = 15.0 Hz, 1H), 1.72 (d, J = 14.6 Hz, 1H), 1.65 - 1.48 (m, 2H), 1.44 (d, J = 2.3 Hz, 6H), 1.11 (s, 3H). LCMS (ESI-MS) m / z = 491.2 [M+H] + .

[0338] Second eluting isomer: 1-(((5S,7S,8R)-3-(5-(2-Hydroxypropan-2-yl)pyrazin-2-yl)-8-methoxy-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile as a white solid (14.8 mg, purity 99.6%, 99.5% ee). 11H NMR (300 MHz, DMSO-d6) δ 9.18 (d, J = 1.5 Hz, 1H), 8.58 (d, J = 1.5 Hz, 1H), 8.37 (s, 1H), 8.25 - 8.20 (m, 1H), 7.84 (d, J = 8.3 Hz, 1H), 7.60 (dd, J = 8.3, 1.5 Hz, 1H), 5.42 (s, 1H), 4.31 (d, J = 14.5 Hz, 1H), 4.21 (d, J = 14.5 Hz, 1H), 3.81 (q, J = 10.3 Hz, 2H), 3.31 (s, 3H), 3.07 - 3.00 (m, 1H), 2.04 (d, J = 9.7 Hz, 2H), 1.95 (d, J = 15.0 Hz, 1H), 1.72 (d, J = 14.6 Hz, 1H), 1.65 - 1.48 (m, 2H), 1.44 (d, J = 2.3 Hz, 6H), 1.11 (s, 3H). LCMS (ESI-MS) m / z = 491.2 [M+H] + .

[0339] Example 11. Preparation of 1-(((5S,7S)-8,8-difluoro-3-neopentyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000181.tif48165

[0340] Detailed procedure rac-1-(((5S,7S)-8,8-difluoro-3-neopentyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000182.tif58165In a mixture of rac-1-(((5S,7S)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (1 g, 2.89 mmol) and 1-iodo-2,2-dimethylpropane (859 mg, 4.34 mmol) in N,N-dimethylformamide (10 mL), sodium hydroxide (347 mg, 8.67 mmol) was added. The reaction mixture was stirred at 100 °C for 3 hours and cooled to room temperature. The mixture was quenched with aqueous ammonium chloride (50 mL), extracted with ethyl acetate (3 × 30 mL), dried over sodium sulfate, filtered, and concentrated to obtain the crude product. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0% - 60% petroleum ether, to give the product rac-1-(((5S,7S)-8,8-difluoro-3-neopentyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (200 mg) as a white solid. LCMS (ESI-MS) m / z = 417.2 [M+H] + .

[0341] 1-(((5S,7S)-8,8-difluoro-3-neopentyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile and 1-(((5R,7R)-8,8-difluoro-3-neopentyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile A mixture of TIFF2025522953000183.tif58165rac-1-(((5S,7S)-8,8-difluoro-3-neopentyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (200 mg, 0.48 mmol) was separated by preparative chiral HPLC under the following conditions: column: CHIRALPAK IH, 2 x 25 cm, 5 μm, mobile phase A: MTBE (0.5% 2M NH3-MeOH)--HPLC, mobile phase B: IPA--HPLC, flow rate: 20 mL / min, gradient: 40% B to 40% B in 12 minutes, wavelength: 220 / 254 nm; RT1 (min): 7.342; RT2 (min): 9.486; sample solvent: EtOH--HPLC, injection volume: 0.3 mL. The desired fractions were combined and lyophilized to give two products.

[0342] The first eluting isomer: 1-(((5S,7S)-8,8-difluoro-3-neopentyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile as a white solid (76.5 mg, purity 99.8%, 100% ee). 1 H NMR (400 MHz, DMSO-d6) δ 8.56 (s, 1H), 8.31 (d, J = 1.5 Hz, 1H), 7.83 (d, J = 8.5 Hz, 1H), 7.61 (dd, J = 8.4, 1.5 Hz, 1H), 4.65 (dd, J = 14.6, 6.1 Hz, 1H), 4.41 (dd, J = 14.6, 7.7 Hz, 1H), 3.46 - 3.37 (m, 2H), 2.90 - 2.71 (m, 3H), 2.15 - 1.93 (m, 3H), 1.88 - 1.63 (m, 3H), 0.82 (s, 9H). LCMS (ESI-MS) m / z = 417.2 [M+H] + .

[0343] Second eluting isomer: 1-(((5R,7R)-8,8-difluoro-3-neopentyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile as a white solid (19.9 mg, purity 99.0%, 98.4% ee). 1 H NMR (400 MHz, DMSO-d6) δ 8.56 (s, 1H), 8.31 (d, J = 1.5 Hz, 1H), 7.83 (d, J = 8.3 Hz, 1H), 7.61 (dd, J = 8.4, 1.5 Hz, 1H), 4.65 (dd, J = 14.7, 6.0 Hz, 1H), 4.41 (dd, J = 14.6, 7.7 Hz, 1H), 3.44-3.35 (m, 2H), 2.91-2.80 (m, 3H), 2.16-1.93 (m, 3H), 1.86-1.70 (m, 3H), 0.82 (s, 9H). LCMS (ESI-MS) m / z = 417.2 [M+H] + .

[0344] Example 12. Preparation of 1-(((5S,7S)-3-(2-chlorophenyl)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000184.tif48165

[0345] Detailed procedure 1-((3-(2-chlorophenyl)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000185.tif581651, In 1,4-dioxane (10 mL), to a diastereomeric mixture of 1-((8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (500 mg, 1.45 mmol) and 1-chloro-2-iodobenzene (519 mg, 2.18 mmol), N1,N2-dimethylethane-1,2-diamine (255 mg, 2.9 mmol), tribasic potassium phosphate (615 mg, 2.9 mmol) and copper(I) iodide (139 mg, 0.73 mmol) were added. The resulting mixture was stirred at 100 °C for 5 h under a nitrogen atmosphere and then cooled to room temperature. The mixture was quenched with aqueous ammonium chloride (50 mL), extracted with ethyl acetate (3 × 30 mL), dried over sodium sulfate, filtered and concentrated to give a crude product. The residue was purified by silica gel column chromatography and then eluted with ethyl acetate in 0% - 55% petroleum ether to give the product 1-((3-(2-chlorophenyl)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (400 mg) as a white solid. LCMS (ESI-MS) m / z = 457.1 [M+H] + .

[0346] rac-1-(((5S,7S)-3-(2-chlorophenyl)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H -benzo[d]imidazole-6-carbonitrile and rac-1-(((5R,7S)-3-(2-chlorophenyl)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5 ]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000186.tif581651 - ((3-(2-Chlorophenyl)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (400 mg, 0.88 mmol), a diastereomeric mixture, was separated by preparative chiral SFC under the following conditions: Column: YMC-Actus Triart Diol-HILIC, 3 x 25 cm, 5 μm, Mobile phase A: C O2 , Mobile phase B: MeOH (0.1% 2M NH3-MeOH), Flow rate: 75 mL / min; Gradient: Isocratic 24% B; Column temperature (°C): 35; Back pressure (bar): 100; Wavelength: 220 nm; RT1 (min): 4.26, Sample solvent: MeOH - HPLC, Injection volume: 3 mL. The desired fractions were combined and lyophilized to give two products.

[0347] The first eluting isomer: rac-1-(((5S,7S)-3-(2-Chlorophenyl)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile as a white solid (240 mg).

[0348] The second eluting isomer: rac-1-(((5R,7S)-3-(2-Chlorophenyl)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile as a white solid (20 mg).

[0349] 1-(((5R,7R)-3-(2-Chlorophenyl)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile and 1-(((5S,7S)-3-(2-Chlorophenyl)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile A mixture of TIFF2025522953000187.tif63165rac-1-(((5S,7S)-3-(2-chlorophenyl)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]dec-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (240 mg, 0.88 mmol) was separated by preparative chiral HPLC under the following conditions: column: CHIRALPAK IF, 2x25 cm, 5 μm, mobile phase A: MTBE (0.5% 2M NH3-MeOH)--HPLC, mobile phase B: EtOH--HPLC, flow rate: 20 mL / min, gradient: 20%B to 20%B in 23 minutes, wavelength: 220 / 254 nm; RT1 (min): 11.816, RT2 (min): 16.483, sample solvent: EtOH--HPLC, injection volume 2 mL. The desired fractions were combined and lyophilized to obtain two products.

[0350] The first eluting isomer: 1-(((5R,7R)-3-(2-chlorophenyl)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]dec-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (10.6 mg, 99.5% purity, 100% ee) was obtained as a white solid. 1 H NMR (400 MHz, DMSO-d6) δ 8.59 (s,1H),8.35 (s,1H),7.85 (d, J =8.4 Hz,1H),7.63 (dd, J = 8.4,1.5 Hz, 1H), 7.60 -7.51 (m, 2H), 7.45-7.36 (m, 2H), 4.70 (dd, J = 14.6, 5.9 Hz, 1H), 4.45 (dd, J = 14.6, 8.0 Hz, 1H), 3.77 -3.66 (m,2H),3.02-2.82 (m,1H),2.26-2.12 (m,3H),2.09-1.93 (m,2H),1.91-1.78 (m,1H). LCMS (ESI-MS) m / z = 457.1 [M+H] + .

[0351] Second eluting isomer: 1-(((5S,7S)-3-(2-chlorophenyl)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile as a white solid (31.6 mg, purity 99.2%, 100% ee). 1 H NMR (400 MHz, DMSO-d6) δ 8.59 (s, 1H), 8.35 (d, J = 1.5 Hz, 1H), 7.85 (d, J = 8.3 Hz, 1H), 7.63 (dd, J = 8.3, 1.5 Hz, 1H), 7.59-7.50 (m, 2H), 7.45-7.34 (m, 2H), 4.70 (dd, J = 14.6, 5.9 Hz, 1H), 4.45 (dd, J = 14.6, 8.0 Hz, 1H), 3.77-3.66 (m, 2H), 3.00-2.84 (m, 1H), 2.27-2.12 (m, 3H), 2.03-1.89 (m, 2H), 1.89-1.78(m, 1H). LCMS (ESI-MS) m / z = 457.1 [M+H] + .

[0352] Example 13. Preparation of 1-(((5S,7S)-3-(2-ethoxyphenyl)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000188.tif53165

[0353] Detailed procedure 1-((3-(2-ethoxyphenyl)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000189.tif581651, In 1,4-dioxane (10 mL), to a diastereomeric mixture of 1-((8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (250 mg, 0.72 mmol) and 1-bromo-2-ethoxybenzene (217 mg, 1.08 mmol), N1,N2-dimethylethane-1,2-diamine (127 mg, 1.44 mmol), tripotassium phosphate tribasic (305 mg, 1.44 mmol) and copper(I) iodide (69 mg, 0.36 mmol) were added. The resulting mixture was stirred at 100 °C for 6 h under a nitrogen atmosphere, cooled to room temperature, quenched with aqueous ammonium chloride (50 mL), and the resulting solution was extracted with ethyl acetate (3 × 30 mL). The organic layer was dried over sodium sulfate, filtered, concentrated to give the crude product, and the residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0% to 58% petroleum ether to give 1-((3-(2-ethoxyphenyl)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (220 mg) as a white solid. LCMS (ESI-MS) m / z = 467.2 [M+H] + .

[0354] rac-1-(((5S,7S)-3-(2-ethoxyphenyl)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H -benzo[d]imidazole-6-carbonitrile and rac-1-(((5R,7S)-3-(2-ethoxyphenyl)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000190.tif631651 - ((3-(2-Ethoxyphenyl)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (220 mg, 0.47 mmol) of the diastereomer mixture was separated by preparative achiral SFC under the following conditions: column: GreenSep Naphthyl, 3 x 25 cm, 5 μm, mobile phase A: CO2, mobile phase B: MeOH (0.1% 2M NH3-MeOH), flow rate: 75 mL / min; gradient: isocratic 25% B; column temperature (°C): 35; back pressure (bar): 100; wavelength: 220 nm; RT1 (min): 5.99, RT2 (min): 6.51, sample solvent: MeOH - HPLC, injection volume: 1 mL. The desired fractions were combined and lyophilized to obtain two products.

[0355] The isomer eluting first: rac-1-(((5R,7S)-3-(2-Ethoxyphenyl)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (30 mg) as a white solid.

[0356] The isomer eluting second: rac-1-(((5S,7S)-3-(2-Ethoxyphenyl)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (140 mg) as a white solid.

[0357] 1-(((5R,7R)-3-(2-Ethoxyphenyl)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile and 1-(((5S,7S)-3-(2-Ethoxyphenyl)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000191.tif58165rac-1-(((5S,7S)-3-(2-chlorophenyl)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (140 mg, 0.3 mmol) was separated by preparative chiral HPLC under the following conditions: column: CHIRALPAK ID, 2 x 25 cm, 5 μm, mobile phase A: MTBE (2 mM NH3-MeOH)--HPLC, mobile phase B: EtOH--HPLC, flow rate: 20 mL / min, gradient: 8% B to 8% B over 31 minutes, wavelength: 220 / 254 nm; RT1 (min): 20.16, RT2 (min): 26.092, sample solvent: EtOH--HPLC, injection volume 0.4 mL. The desired fractions were combined and lyophilized to give two products.

[0358] The first eluting isomer: 1-(((5R,7R)-3-(2-ethoxyphenyl)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile as a white solid (18.0 mg, 98.9% purity, 100% ee). 1 H NMR (400 MHz, DMSO-d6) δ 8.59 (s, 1H), 8.34 (s, 1H), 7.85 (d, J = 8.4 Hz, 1H), 7.63 (dd, J = 8.4, 1.5 Hz, 1H), 7.32 - 7.25 (m, 2H), 7.06 (d, J = 8.4 Hz, 1H), 6.94 (t, J = 7.6 Hz, 1H), 4.69 (dd, J = 14.7, 5.9 Hz, 1H), 4.44 (dd, J = 14.6, 8.0 Hz, 1H), 4.04 - 3.93 (m, 2H), 3.64 (s, 2H), 3.01 - 2.84 (m, 1H), 2.23 - 2.08 (m, 3H), 1.96 - 1.78 (m, 3H), 1.17 (t, J = 6.9 Hz, 3H). LCMS (ESI-MS) m / z = 467.2 [M+H] + .

[0359] Second eluting isomer: 1-(((5S,7S)-3-(2-ethoxyphenyl)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile as a white solid (40.4 mg, purity 99.8%, 100% ee). 1 H NMR (400 MHz, DMSO-d6) δ 8.59 (s, 1H), 8.34 (s, 1H), 7.85 (d, J = 8.4 Hz, 1H), 7.62 (dd, J = 8.4, 1.5 Hz, 1H), 7.31 - 7.24 (m, 2H), 7.10 - 7.04 (m, 1H), 6.94 (td, J = 7.7, 1.3 Hz, 1H), 4.69 (dd, J = 14.6, 5.9 Hz, 1H), 4.44 (dd, J = 14.6, 8.1 Hz, 1H), 4.01 - 3.92 (m, 2H), 3.64 (s, 2H), 3.00 - 2.82 (m, 1H), 2.23 - 2.12 (m, 3H), 1.96 - 1.82 (m, 3H), 1.17 (t, J = 6.9 Hz, 3H). LCMS (ESI-MS) m / z = 467.2 [M+H] + .

[0360] Example 14. Preparation of 1-(((5S,7S)-3-(5-(tert-butyl)pyrazin-2-yl)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000192.tif58165

[0361] Detailed procedure 2-Bromo-N-(3,3-dimethyl-2-oxobutyl)acetamide In chloroform (30 mL), to a mixture of calcium carbonate (7.15 g, 93.8 mmol), 1-amino-3,3-dimethylbutan-2-one hydrochloride (1.8 g, 15.6 mmol) in water (10 mL) was added at 5 °C under a nitrogen atmosphere. Subsequently, 2-bromoacetyl bromide (5.90 g, 39.1 mmol) was added at 5 °C. The resulting mixture was warmed to room temperature and stirred for 6 h. The reaction was diluted with aqueous sodium hydrogen carbonate (30 mL) and filtered. The organic layer was dried over sodium sulfate, filtered, and concentrated to give the crude product 2-bromo-N-(3,3-dimethyl-2-oxobutyl)acetamide (2.4 g) as a yellow oil. LCMS (ESI-MS) m / z = 236.0 [M+H] + .

[0362] 5-(tert-Butyl)pyrazin-2-ol In ammonia (20 ml, 7N in MeOH), to a mixture of 2-bromo-N-(3,3-dimethyl-2-oxobutyl)acetamide (2.3 g, 9.74 mmol) was added sodium iodide (0.26 g, 1.75 mmol). The resulting mixture was stirred at room temperature for 3 days. The mixture was quenched with aqueous ammonium chloride (50 mL), extracted with ethyl acetate (3 × 30 mL), dried over sodium sulfate, filtered, and concentrated to give the crude product 5-(tert-butyl)pyrazin-2-ol (740 mg) as a yellow oil. LCMS (ESI-MS) m / z = 153.1 [M+H] + .

[0363] 5-(tert-Butyl)pyrazin-2-yl trifluoromethanesulfonate In dichloromethane (10 mL), to a mixture of 5-tert-butylpyrazin-2-ol (730 mg, 4.80 mmol) was added triethylamine (1.46 g, 14.4 mmol) and trifluoromethanesulfonic anhydride (2.03 g, 7.19 mmol) at 0 °C under a nitrogen atmosphere. The mixture was warmed to room temperature and stirred for 6 hours. The mixture was quenched with water (50 mL), extracted with ethyl acetate (3 × 30 mL), dried over sodium sulfate, filtered, and concentrated to obtain a crude product. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0% - 30% petroleum ether to give the product 5-(tert-butyl)pyrazin-2-yl trifluoromethanesulfonate (500 mg) as a yellow oil. LCMS (ESI-MS) m / z = 285.0 [M+H] + .

[0364] 1-((3-(5-(tert-Butyl)pyrazin-2-yl)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000196.tif631651,4-Dioxane (10 mL), a diastereomeric mixture of 1-((8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (600 mg, 1.73 mmol) and 5-(tert-butyl)pyrazin-2-yl trifluoromethanesulfonate (738 mg, 2.60 mmol), was added to BrettPhos (93 mg, 0.173 mmol), BrettPhos Pd G3 (157 mg, 0.173 mmol) and cesium carbonate (1.69 g, 5.19 mmol). The resulting mixture was stirred at 90 °C overnight under a nitrogen atmosphere and cooled to room temperature. The reaction was quenched with aqueous ammonium chloride (50 mL), extracted with ethyl acetate (3 × 30 mL), dried over sodium sulfate, filtered and concentrated to give the crude product. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0% - 70% petroleum ether to give the product 1-((3-(5-(tert-butyl)pyrazin-2-yl)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (420 mg) as a pale yellow solid. LCMS (ESI-MS) m / z = 481.2 [M+H] + .

[0365] rac-1-(((5R,7S)-3-(5-(tert-butyl)pyrazin-2-yl)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile and rac-1-(((5S,7S)-3-(5-(tert-butyl)pyrazin-2-yl)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000197.tif731651 - ((3 - (5 - (tert - butyl)pyrazin - 2 - yl) - 8,8 - difluoro - 2 - oxo - 1 - oxa - 3 - azaspiro[4.5]decane - 7 - yl)methyl) - 1H - benzo[d]imidazole - 6 - carbonitrile (420 mg, 0.87 mmol), a diastereomeric mixture, was separated by preparative chiral SFC under the following conditions: column: DAICEL DCpak P4VP, 3 x 25 cm, 5 μm; mobile phase A: CO2; mobile phase B: MeOH (0.1% 2M NH3 - MeOH); flow rate: 60 mL / min; gradient: isocratic 17% B; column temperature (25 °C): 35; back pressure (bar): 100; wavelength: 254 nm; RT1 (min): 7.62, RT2 (min): 9; sample solvent: MeOH - HPLC; injection volume: 2.5 mL. The desired fractions were combined and lyophilized to give two products.

[0366] The isomer eluting first: rac - 1 - (((5R,7S) - 3 - (5 - (tert - butyl)pyrazin - 2 - yl) - 8,8 - difluoro - 2 - oxo - 1 - oxa - 3 - azaspiro[4.5]decane - 7 - yl)methyl) - 1H - benzo[d]imidazole - 6 - carbonitrile (300 mg) was obtained as a white solid.

[0367] The isomer eluting second: rac - 1 - (((5S,7S) - 3 - (5 - (tert - butyl)pyrazin - 2 - yl) - 8,8 - difluoro - 2 - oxo - 1 - oxa - 3 - azaspiro[4.5]decane - 7 - yl)methyl) - 1H - benzo[d]imidazole - 6 - carbonitrile (50 mg) was obtained as a white solid.

[0368] 1-(((5S,7S)-3-(5-(tert-Butyl)pyrazin-2-yl)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile and 1-(((5R,7R)-3-(5-(tert-Butyl)pyrazin-2-yl)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile A mixture of TIFF2025522953000198.tif68165rac-1-(((5S,7S)-3-(5-(tert-butyl)pyrazin-2-yl)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (300 mg, 0.62 mmol) was separated by preparative chiral HPLC under the following conditions: column: CHIRALPAK IE, 2 x 25 cm, 5 μm, mobile phase A: MTBE (0.5% 2M NH3-MeOH)--HPLC, mobile phase B: EtOH--HPLC, flow rate: 16 mL / min, gradient: 50%B - 50%B in 12 minutes, wavelength: 220 / 254 nm; RT1 (min): 7.528, RT2 (min): 9.869, sample solvent: EtOH, injection volume: 0.4 mL. The desired fractions were combined and lyophilized to obtain two products.

[0369] The first eluting isomer: 1-(((5S,7S)-3-(5-(tert-Butyl)pyrazin-2-yl)-8,8-difluoro-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile as a white solid (143.5 mg, purity 98.6%, 100% ee). 11H NMR (400 MHz, DMSO-d6) δ 9.20 (d, J = 1.6 Hz, 1H), 8.56 (s, 1H), 8.45 (d, J = 1.5 Hz, 1H), 8.30 (d, J = 1.4 Hz, 1H), 7.83 (d, J = 8.3 Hz, 1H), 7.61 (dd, J = 8.4, 1.5 Hz, 1H), 4.72 - 4.64 (m, 1H), 4.40 (dd, J = 14.7, 7.7) Hz, 1H), 3.91 (q, J = 10.4 Hz, 2H), 3.00 - 2.80 (m, 1H), 2.26 - 2.02 (m, 4H), 2.01 - 1.89 (m, 2H), 1.32 (s, 9H). LCMS (ESI-MS) m / z = 481.2 [M+H] +

[0370] Second eluting isomer: 1 - (((5R,7R)-3-(5-(tert - butyl)pyrazin - 2 - yl)-8,8 - difluoro - 2 - oxo - 1 - oxa - 3 - azaspiro[4.5]decane - 7 - yl)methyl)-1H - benzo[d]imidazole - 6 - carbonitrile as a white solid (51.7 mg, purity 99.8%, 100% ee). 1 1H NMR (400 MHz, DMSO-d6) δ 9.20 (d, J = 1.6 Hz, 1H), 8.56 (s, 1H), 8.45 (d, J = 1.6 Hz, 1H), 8.35 - 8.28 (m, 1H), 7.83 (d, J = 8.4 Hz, 1H), 7.61 (dd, J = 8.4, 1.5 Hz, 1H), 4.67 (dd, J = 14.7, 5.9 Hz, 1H), 4.40 (dd, J = 14.7), 7.7 Hz, 1H), 3.91 (q, J = 10.4 Hz, 2H), 3.02 - 2.82 (m, 1H), 2.24 - 2.04 (m, 4H), 1.99 - 1.83 (m, 2H), 1.32 (s, 9H). LCMS (ESI-MS) m / z = 481.2 [M+H] + .

[0371] Example 15. Preparation of 1-(((5S,7S)-8,8-difluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000199.tif58165

[0372] Detailed procedure 4,4-difluoro-3-methylcyclohexa-2,5-dien-1-one TIFF2025522953000200.tif32165 A solution of 4-fluoro-3-methylphenol (25.2 g, 1 equivalent, 200 mmol) in DCM (1.5 L) at room temperature was added dropwise with pyridine hydrofluoride (20 mL, 70% by weight, 0.78 equivalent, 156 mmol) over 10 minutes or more. Subsequently, phenyl-λ3-iodanediyl diacetate (77.3 g, 1.2 equivalents, 240 mmol) was added all at once, and the reaction mixture was stirred at room temperature for 15 minutes. Potassium carbonate (138 g, 5 equivalents, 1.00 mol) was added, and the resulting mixture was stirred under the same conditions for 10 minutes. The mixture was filtered, and the filtrate was washed with 1 M HCl (3x) and brine (1x), dried over Na2SO4, filtered, and concentrated under reduced pressure (45 °C, not less than 850 mbar because the product is volatile). It was reduced to about 200 mL. The crude solution was used in the next step without further purification. 1 Obtained by 1H NMR (400 MHz, CDCl3) δ 6.80 (dt, J = 10.2, 5.5 Hz, 1H), 6.32 - 6.26 (m, 1H), 6.15 - 6.09 (m, J = 1.5 Hz, 1H), 2.09 (m, J = 1.5 Hz, 1H), 2.09 (m, J = 10.2, 5.5 Hz, 1H), dt, J = 1.9, 1.0 Hz, 3H). 19 19F NMR (376 MHz, CDCl3) δ -102.58.

[0373] 4,4-difluoro-3-methylcyclohexa-2-en-1-one In TIFF2025522953000201.tif42165tert-butyl methyl ether (1.33 L), crude 4,4-difluoro-3-methylcyclohexa-2,5-dien-1-one (28.8 g, 1 equivalent, 200 mmol), 3,5-pyridinedicarboxylic acid, 1,4-dihydro-2,6-dimethyl-diester (HEH; 111 g, 2.2 equivalents, 440 mmol), were mechanically stirred at 60 °C overnight in the presence of silicon dioxide (400 g, 33.3 equivalents, 6.66 mol) until refluxed. Next, since there was no complete conversion (aliquot crude 19 by FNMR), 0.2 equivalent of HEH was added and the mixture was stirred at 60 °C for an additional 5 hours. The mixture was cooled to room temperature, filtered to remove silica, and the volume of the filtrate was reduced to 400 mL on a rotary evaporator. 1.6 L of pentane was added, the mixture was filtered through a silica pad, concentrated to 200 mL (fraction A), the pad was washed with 1 L of pentane:MTBE 4:1 and concentrated to 250 mL (fraction B) (Note: Most of the unreacted HHH can be removed through this filtration). NMR showed that there were still impurities of HEH in both fractions stored overnight in the freezer. Both fractions had precipitated HEH filtered off. Each of the fractions was washed with 5% aqueous CuSO4 (x2) (Note: This was done to remove any residue of HEH) and 3M HCl (×3) (Note: This is necessary to remove Hunig's pyridine). Subsequently, qNMR showed that 4,4-difluoro-3-methylcyclohex-2-en-1-one (14.6 g, 100 mmol, 50%) was obtained as a solution in MTBE and was used in the next step without further purification. 1 1H NMR (400 MHz, CDCl3) δ 6.00 (h, J = 1.5 Hz, 1H), 2.63 (dd, J = 7.2, 6.0 Hz, 2H), 2.52 - 2.41 (m, 2H), 2.05 (dt, J = 1.5, 0.8 Hz, 3H). 19 19F NMR (376 MHz, CDCl3) δ -100.50 (t, J = 13.3 Hz).

[0374] (S)-4,4-Difluoro-3-methyl-3-(nitromethyl)cyclohexan-1-one In a flask of TIFF2025522953000202.tif371651L, a solution of crude 4,4-difluoro-3-methylcyclohex-2-en-1-one (7.98 g, 210 mL, 0.26 molar concentration, 1 equivalent, 54.6 mmol) in MTBE was diluted to a total volume of MTBE (546 mL). 1-((1S,2S)-2-Aminocyclohexyl)-3-(3,5-bis(trifluoromethyl)phenyl)thiourea (2.10 g, 0.1 equivalent, 5.46 mmol) and nitromethane (33.3 g, 29.4 mL, 10 equivalents, 546 mmol) were added, and the mixture was stirred at 50 °C under N2 for 2 days. The flask was cooled to room temperature. The mixture was washed with saturated KHSO4 solution (3x), saturated NaHCO3 solution (3x), H2O (1x) and brine (1x). Dried (Na2SO4), filtered and concentrated. The crude product was purified by silica chromatography (220 g) using a gradient of 0 - 25% EtOAc in c-hexane (2 CV 0%, 10 CV 0 - 10%, 2 CV 10%, 10 CV 10 - 20%, 4 CV 20 - 25%, 2 CV 25%) to give (S)-4,4-difluoro-3-methyl-3-(nitromethyl)cyclohexan-1-one (4.11 g, 19.8 mmol, 36.3%) as a yellow oil. 1 H NMR (400 MHz, CDCl3) δ 4.63 (d, J = 11.1 Hz, 1H), 4.58 (d, J = 11.1 Hz, 1H), 2.88 - 2.80 (m, 1H), 2.72 - 2.62 (m, 1H), 2.56 - 2.47 (m, 2H), 2.45 - 2.30 (m, 2H), 1.28 (d, J = 1.0 Hz, 3H). 19 F NMR (376 MHz, CDCl3) δ -110.17 (dd, J = 24.0, 12.9 Hz), -110.31 (td, J = 10.4, 3.5 Hz).

[0375] (S)-8,8-Difluoro-7-methyl-7-(nitromethyl)-1,4-dioxaspiro[4.5]decane TIFF2025522953000203.tif32165 A solution of (S)-4,4-difluoro-3-methyl-3-(nitromethyl)cyclohexan-1-one (4.0 g, 1 equivalent, 19.3 mmol) in anhydrous DCM (3.8 M) was added with ethylene glycol (1.59 mL, 1.48 equivalents, 28.6 mmol) and trimethyl orthoformate (3.2 mL, 1.5 equivalents, 29.0 mmol). The resulting reaction mixture was stirred at room temperature for 5 minutes and then cooled to 0 °C in an ice bath. Methanesulfonic acid (186 μL, 0.148 equivalents, 2.86 mmol) was added dropwise to this mixture. The reaction mixture was removed from the ice bath, warmed to room temperature, and stirred overnight. The reaction was cooled back to 0 °C, quenched with 100 mL of water, and partitioned with 100 mL of DCM. The layers were separated and the aqueous layer was extracted with DCM (3 x 50 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated to give (S)-8,8-difluoro-7-methyl-7-(nitromethyl)-1,4-dioxaspiro[4.5]decane (4.2 g, 87%) as a clear orange-yellowish oil. 1 1H NMR (400 MHz, CDCl3) δ 4.82 (d, J = 11.0 Hz, 1H), 4.51 (d, J = 11.1 Hz, 1H), 4.02 - 3.92 (m, 4H), 2.29 - 2.15 (m, 1H), 2.15 - 2.09 (m, 1H), 2.08 - 1.95 (m, 1H), 1.91 - 1.79 (m, 3H), 1.30 (d, J = 1.4 Hz, 3H). 19 19F NMR (376 MHz, CDCl3) δ -109.08 (dddd, J = 244.3, 20.7, 7.2 Hz), -110.28 (dddd, J = 243.9, 21.2, 7.6, 3.0 Hz).

[0376] (S)-(8,8-Difluoro-7-methyl-1,4-dioxaspiro[4.5]decane-7-yl)methanamine TIFF2025522953000204.tif32165 An aqueous solution of (S)-8,8-difluoro-7-methyl-7-(nitromethyl)-1,4-dioxaspiro[4.5]decane (1 equiv) in EtOAc was added to Pd (10% in charcoal; 0.075 equiv) under an inert atmosphere. MeOH (MeOH:EtOAc 2:1, 0.1 M) was added to this mixture, the reaction vessel was emptied, and the balloon was filled with H2. The reaction mixture was stirred at room temperature overnight. Complete conversion was observed by TLC (TLC 5% MeOH / DCM shows complete conversion using ninhydrin as the staining solution). The reaction vessel was emptied and filled with nitrogen. The reaction contents were filtered through celite, washed with EtOAc, and concentrated in vacuo to give (S)-(8,8-difluoro-7-methyl-1,4-dioxaspiro[4.5]decane-7-yl)methanamine (3.7 g, quant.) as a pale yellow transparent oil, which was used in the next step without further purification. 1 1H NMR (400 MHz, CDCl3) δ 3.97 - 3.91 (m, 4H), 2.87 (d, J = 13.3 Hz, 1H), 2.77 (d, J = 13.4 Hz, 1H), 2.13 - 2.01 (m, 2H), 1.84 - 1.79 (m, 3H), 1.64 - 1.58 (m, 1H), 1.12 (d, J = 1.5 Hz, 3H). 19 19F NMR (376 MHz, CDCl3) δ -106.57 - -107.45 (m), -111.32 - -112.12 (m).

[0377] (S)-3-(((8,8-Difluoro-7-methyl-1,4-dioxaspiro[4.5]decane-7-yl)methyl)amino)-4-nitrobenzonitrile (S)-(8,8-Difluoro-7-methyl-1,4-dioxaspiro[4.5]decan-7-yl)methanamine (1.00 equiv) was dissolved in MeCN (0.35 M). K2CO3 (2.00 equiv) and 3-fluoro-4-nitrobenzonitrile (1.00 equiv) were added, and the resulting orange mixture was stirred at room temperature for 24 h. The reaction mixture was filtered through a frit, and the filtrate was concentrated and purified on a silica gel column (0 - 1% MeOH / DCM) to afford (S)-3-(((8,8-difluoro-7-methyl-1,4-dioxaspiro[4.5]decan-7-yl)methyl)amino)-4-nitrobenzonitrile (5.4 g, 88%). 1 1H NMR (400 MHz, CDCl3) δ 8.41 (s, 1H), 8.30 - 8.22 (m, 1H), 6.85 (dd, J = 8.7, 1.6 Hz, 1H), 4.07 - 3.90 (m, 4H), 3.56 - 3.44 (m, 2H), 2.22 - 2.09 (m, 2H), 1.93 - 1.82 (m, 3H), 1.76 (dd, J = 14.0, 2.6 Hz, 1H), 1.30 - 1.18 (m, 4H). 19 19F NMR (376 MHz, CDCl3) δ -107.02~-107.96 (m), -110.15~-111.12 (m).

[0378] (S)-4-Amino-3-(((8,8-difluoro-7-methyl-1,4-dioxaspiro[4.5]decan-7-yl)methyl)amino)benzonitrile In an inert atmosphere, a solution of (S)-3-(((8,8-difluoro-7-methyl-1,4-dioxaspiro[4.5]decan-7-yl)methyl)amino)-4-nitrobenzonitrile (1 eq) in ethyl acetate was added to Pd (10% on charcoal; 0.05 eq). To this mixture, MeOH (MeOH:AcOEt 2:1, 0.1 M) was added, the reaction vessel was emptied, and the balloon was filled with H2. The reaction mixture was stirred at room temperature for 2 h by TLC (TLC: 100% DCM, ninhydrin as the staining agent) until complete conversion was observed. The reaction vessel was emptied and filled with nitrogen. The reaction contents were filtered through celite, washed with EtOAc, and concentrated in vacuo to give (S)-4-amino-3-(((8,8-difluoro-7-methyl-1,4-dioxaspiro[4.5]decan-7-yl)methyl)amino)benzonitrile (4.9 g, 99%). 1 1H NMR (400 MHz, CDCl3) δ 7.02 (dd, J = 8.0, 1.8 Hz, 1H), 6.89 (d, J = 1.8 Hz, 1H), 6.67 (d, J = 7.9 Hz, 1H), 4.02 - 3.91 (m, 4H), 3.22 - 3.15 (m, 2H), 2.25 - 2.08 (m, 2H), 1.95 (dd, J = 14.2, 2.7 Hz, 1H), 1.85 (d, J = 6.5 Hz, 2H), 1.73 (dt, J = 14.2, 2.2 Hz, 1H), 1.27 (d, J = 1.5 Hz, 3H). 19 19F NMR (376 MHz, CDCl3) δ -105.99 - -106.88 (m), -109.87 (dddd, J = 242.6, 19.3, 10.3 Hz).

[0379] (S)-1-((8,8-difluoro-7-methyl-1,4-dioxaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000207.tif53165(S)-4-Amino-3-(((8,8-difluoro-7-methyl-1,4-dioxaspiro[4.5]decane-7-yl)methyl)amino)benzonitrile (4.9 g, 1 equivalent, 15 mmol) was added to trimethyl orthoformate (46 mL, 29 equivalents, 420 mmol), followed by formic acid (0.56 mL, 1 equivalent, 15 mmol). The resulting mixture was stirred at room temperature. After the start of the reaction, additional formic acid (0.56 mL, 1 equivalent, 15 mmol) was added at t = 90 min, 120 min, and 180 min, and then the mixture was stirred overnight. The reaction contents were partitioned between 500 mL of EtOAc and 300 mL of saturated aqueous NaHCO3, and the layers were separated. The aqueous layer was back-extracted with EtOAc (3 x 150 mL). The combined organics were dried over anhydrous Na2SO4, filtered, and concentrated under vacuum. The crude material was purified by silica gel chromatography (80 g of SiO2, 0 - 70% EtOAc:hexane) to give (S)-1-((8,8-difluoro-7-methyl-1,4-dioxaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (4.4 g, 13 mmol, 87%) as a white solid. 1 1H NMR (400 MHz, CDCl3) δ 8.17 (s, 1H), 7.86 (dd, J = 8.4, 0.7 Hz, 1H), 7.80 (d, J = 1.5 Hz, 1H), 7.54 (dd, J = 8.4, 1.5 Hz, 1H), 4.47 (d, J = 14.8 Hz, 1H), 4.31 (d, J = 14.9 Hz, 1H), 4.04 ‐3.90 (m, 4H), 2.29 ‐2.07 (m, 2H), 1.97 ‐1.82 (m, 3H), 1.73 ‐1.65 (m, 1H), 1.13 ‐1.09 (m, 3H). 19 19F NMR (376 MHz, CDCl3) δ -107.43 ‐ -108.46 (m), -110.25 (ddd, J = 243.6, 22.9, 7.7 Hz).

[0380] (S)-1-((2,2-difluoro-1-methyl-5-oxocyclohexyl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000208.tifTo a solution of iron(III) chloride (3.27 g, 3.5 equiv, 20.2 mmol) and water (2.18 g, 2.18 mL, 21 equiv, 121 mmol) in CH2Cl2 (19.2 mL) was added a solution of (S)-1-((8,8-difluoro-7-methyl-1,4-dioxaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (2.00 g, 1 equiv, 5.76 mmol) at room temperature. The resulting yellow to amber suspension was heated at 50 °C and iron(III) chloride (3.27 g, 3.5 equiv, 20.2 mmol) in water (2.18 g, 2.18 mL, 21 equiv, 121 mmol) was added at t = 1, 2, and 3 h while maintaining the temperature at 50 °C throughout the process. The reaction was quenched by adding saturated aqueous NaHCO3. The aqueous layer was extracted three times with CH2Cl2, the combined organics were washed with brine, dried over Na2SO4, and concentrated under reduced pressure. NMR indicated that the conversion was not complete. The reaction was restarted under the same conditions as before and iron(III) chloride (3.27 g, 3.5 equiv, 20.2 mmol) in water (2.18 g, 2.18 mL, 21 equiv, 121 mmol) was added at t = 2, 16, 19, and 25 h. The conversion was not complete (70%). The reaction was quenched by adding saturated aqueous NaHCO3. The aqueous layer was extracted three times with CH2Cl2, the combined organics were washed with brine, dried over Na2SO4, and concentrated under reduced pressure. This time, ferric chloride hexahydrate (5.4 g, 3.0 mL, 3.5 equiv, 20 mmol) was added to restart the reaction. The reaction was heated for 2 h, then more ferric chloride hexahydrate (5.4 g, 3.0 mL, 3.5 equiv, 20 mmol) was added and complete conversion was reached after 1 h (determined by LCMS analysis). The reaction was quenched by adding saturated aqueous NaHCO3.The aqueous layer was extracted three times with CH2Cl2. The combined organic matter was washed with brine, dried over Na2SO4, and concentrated under reduced pressure to obtain (S)-1-((2,2-difluoro-1-methyl-5-oxocyclohexyl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (993 mg, 3.27 mmol, 57%) as a light brown foam. 1 1H NMR (400 MHz, CDCl3) δ 8.07 (s, 1H), 7.89 (d, J = 8.4 Hz, 1H), 7.75 (s, 1H), 7.56 (dd, J = 8.3, 1.4 Hz, 1H), 4.38 (q, J = 15.3 Hz, 2H), 2.77 - 2.62 (m, 2H), 2.54 (d, J = 15.7 Hz, 1H), 2.50 - 2.32 (m, 2H), 2.25 (d, J = 14.0 Hz, 1H), 1.15 (s, 3H). 19 19F NMR (376 MHz, CDCl3) δ -109.71 - -109.83 (m), -109.83 - -109.94 (m).

[0381] (S)-1-((2,2-Difluoro-1-methyl-5-methylenecyclohexyl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000209.tif32165N2Under an atmosphere of N2, NaH (60% in mineral oil) (183 mg, 4.58 mmol, 1.4 equiv) was added portionwise to a stirred suspension of methyltriphenylphosphonium bromide (1.64 g, 4.58 mmol, 1.4 equiv) in anhydrous THF (14.6 mL) at 0 °C. The mixture was stirred at 0 °C for 5 h, then warmed to room temperature and stirred at that temperature for 1 h. A solution of (S)-1-((2,2-difluoro-1-methyl-5-oxocyclohexyl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (0.993 g, 3.27 mmol) in anhydrous THF (7.28 mL) was added slowly at 0 °C and allowed to reach room temperature overnight. TLC (CyH / EtOAc: 1 / 1) indicated complete conversion. The reaction mixture was concentrated under reduced pressure, the residue was quenched with saturated aqueous NH4Cl and extracted three times with EtOAc. The combined organic phases were washed with H2O and brine, dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (20 g of SiO2, 10 - 100% EtOAc in cHex) to give (S)-1-((2,2-difluoro-1-methyl-5-methylenecyclohexyl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (473 mg, 3.27 mmol, 48%) as a white crystalline solid. LC-MS: m / z: 302 [M+H] + 11H NMR (400 MHz, CDCl3) δ 8.08 (s, 1H), 7.87 (d, J = 8.3 Hz, 1H), 7.81 ‐7.77 (m, 1H), 7.54 (dd, J = 8.3, 1.5 Hz, 1H), 4.92 (p, J = 1.2 Hz, 1H), 4.76 (t, J = 1.7 Hz, 1H), 4.42 (d, J = 15.1 Hz, 1H), 4.29 (d, J = 15.1 Hz, 1H), 2.44 ‐2.33 (m, 3H), 2.15 ‐2.05 (m, 2H), 2.02 ‐1.91 (m, 2H), 1.42 (d, J = 1.3 Hz, 1H), 1.34 (d, J = 15.1 Hz, 1H), 1.02 (s, 4H), 0.95 (dd, J = 8.1, 6.2 Hz, 1H). 19 19F NMR (376 MHz, CDCl3) δ -105.09 ‐-105.83 (m), -105.86 ‐-106.80 (m), -108.95 (dd, J = 32.0, 11.1 Hz), -109.60 (dd, J = 31.7, 11.4 Hz).

[0382] 1-(((3R,5S)-6,6-difluoro-5-methyl-1-oxaspiro[2.5]octan-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile and 1 -(((3S,5S)-6,6-difluoro-5-methyl-1-oxaspiro[2.5]octan-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000210.tif32165DCM (7.2 mL), at 0 °C, to an aqueous solution of (S)-1-((2,2-difluoro-1-methyl-5-methylenecyclohexyl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (327 mg, 1.09 mmol, 1 equiv) was added m-CPBA (340 mg, 1.52 mmol, 1.4 equiv, 77% wt). The mixture was warmed to room temperature and stirred for 4 h. LCMS indicated that the reaction was not complete, then m-CPBA (340 mg, 1.52 mmol, 1.4 equiv, 77% wt) was added to the reaction mixture and it was stirred at room temperature overnight. The reaction mixture was quenched with a 1 / 1 mixture of saturated aqueous Na2S2O3 / saturated aqueous NaHCO3 and extracted 3 times with DCM. The combined organic layers were washed with saturated NaHCO3 and brine, dried (Na2SO4), filtered and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (40 g of SiO2, 10 CV with DCM / EtOAc 100 / 0 to 85 / 15, then 20 CV with 85 / 15, then up to 75 / 25) to afford the first diastereomer (undesired) 1-(((3R,5S)-6,6-difluoro-5-methyl-1-oxaspiro[2.5]octan-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (96 mg, 0.3 mmol, 28%), and the second diastereomer (desired) 1-(((3S,5S)-6,6-difluoro-5-methyl-1-oxaspiro[2.5]octan-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (64 mg, 0.2 mmol, 19%). LC-MS: m / z: 318 [M+H] + 11H NMR (400 MHz, CDCl3) δ 8.11 (s, 1H), 7.88 (d, J = 8.4 Hz, 1H), 7.76 (d, J = 1.3 Hz, 1H), 7.60 ‐7.54 (m, 1H), 4.42 (d, J = 15.2 Hz, 1H), 4.30 (d, J = 15.2 Hz, 1H), 2.61 (t, J = 4.3 Hz, 1H), 2.57 (d, J = 4.4 Hz, 1H), 2.38 ‐2.22 (m, 4H), 1.37 (dt, J = 11.8, 2.6 Hz, 2H), 1.30 (s, 3H), 1.07 (ddd, J = 13.8, 5.4, 2.6 Hz, 1H).

[0383] 1-(((5S,7S)-8,8-difluoro-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile Lithium 421652-methyl-2-propanoate (0.51 mL, 1.0 molar concentration in THF, 0.51 mmol, 2.7 equiv) was added to a solution of ethyl aminoformate (0.23 g, 2.6 mmol, 13.5 equiv) in NMP (1.3 mL). After stirring at room temperature for 10 minutes, a solution of -(((3S,5S)-6,6-difluoro-5-methyl-1-oxaspiro[2.5]octan-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (60 mg, 0.46 mmol, 1.00 equiv) in NMP (1.3 mL) was added dropwise. Subsequently, the reaction mixture was heated to 100 °C and stirred at that temperature for 21 hours until completely converted by TLC (100% DCM). The reaction mixture was cooled to room temperature, poured into H2O and extracted with EtOAc (x4). The combined organic extracts were washed with NH4Cl (x3), H2O and brine, dried, filtered and concentrated under reduced pressure. The crude product was purified by silica chromatography (4 g of SiO2, 0 - 7% MeOH in DCM) to give 1-(((5S,7S)-8,8-difluoro-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (48 mg, 0.13 mmol, 70%) as a viscous colorless oil. LC-MS: m / z : 361 [M+H] +

[0384] 1-(((5S,7S)-8,8-difluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile In a sealed tube, to 2-(5-chloropyrazin-2-yl)propan-2-ol (29 mg, 0.17 mmol, 1.25 equiv), potassium phosphate tribasic (57 mg, 0.27 mmol, 2.0 equiv), 1-(((5S,7S)-8,8-difluoro-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (48 mg, 0.13 mmol, 1 equiv), 1,4-dioxane (0.67 mL), methyl[2-(methylamino)ethyl]amine (12.0 mg, 0.28 mmol, 1.00 equiv) and CuI (26.9 mg, 0.14 mmol, 0.50 equiv) were added. The reaction mixture was placed under a nitrogen atmosphere and heated at 100 °C for 32 h. The reaction mixture was cooled to room temperature, diluted with DCM, water, and 7 M NH3 in MeOH, and stirred for 10 min. The organic layer was separated and the aqueous layer was extracted with DCM (×2). The combined organic extracts were washed with water (2x), dried, filtered, and concentrated under reduced pressure to give an orange oil. The residue was purified by FCC (0–10% MeOH in DCM) and concentrated to give 1-(((5S,7S)-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-8,8-difluoro-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (26 mg, 0.13 mmol, 39%) as an off-white solid. LC-MS: m / z: 497 [M+H] + 11H NMR (400 MHz, CDCl3) δ 9.37 (d, J = 1.6 Hz, 1H), 8.43 (d, J = 1.6 Hz, 1H), 8.37 (s, 1H), 7.90 (dd, J = 8.4, 0.7 Hz, 1H), 7.80 (d, J = 1.3 Hz, 1H), 7.57 (dd, J = 8.4, 1.4 Hz, 1H), 4.46 (d, J = 15.2 Hz, 1H), 4.35 (d, J = 15.2 Hz, 1H), 3.99 (d, J = 10.8 Hz, 1H), 3.91 (d, J = 10.8 Hz, 1H), 2.45 (dddd, J = 35.7, 18.0, 9.0, 3.9 Hz, 1H), 2.30 ‐2.10 (m, 2H), 2.09 ‐1.93 (m, 3H), 1.63 ‐1.57 (m, 6H), 1.43 (s, 3H). 19 19F NMR (376 MHz, CDCl3) δ -107.42 (d, J = 242.5 Hz), -109.03 (ddd, J = 242.8, 35.6, 10.3 Hz).

[0385] Example 16. Preparation of 1-(((5S,7R)-3-(5-(2-Hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-8-methylene-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000213.tif58165

[0386] Detailed procedure rac-1-(((5S,7S)-3-(5-(2-Hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2,8-dioxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile Copper(I) iodide (281.87 mg, 1.48 mmol) was added to a solution of rac-1-(((5S,7S)-7-methyl-2,8-dioxo-1-oxa-3-azaspiro[4.5]dec-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (500 mg, 1.48 mmol), 2-(5-chloropyrazin-2-yl)propan-2-ol (254.43 mg, 1.48 mmol, 1 equiv), N 1 ,N 2 -dimethylethane-1,2-diamine (260.35 mg, 2.96 mmol) and tripotassium phosphate (628.3 mg, 2.96 mmol) in 1,4-dioxane (5 mL) under a nitrogen atmosphere. The resulting mixture was heated to 100 °C and stirred overnight. The reaction mixture was cooled to room temperature, filtered, and the filtrate was concentrated under vacuum. The residue was purified by silica gel column chromatography, eluting with methanol in dichloromethane from 0% to 3% to give the desired product rac-1-(((5S,7S)-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2,8-dioxo-1-oxa-3-azaspiro[4.5]dec-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (300 mg) as a pale yellow solid. LCMS (ESI-MS) m / z = 475 [M+H] + . 11H NMR (400 MHz, DMSO-d6) δ(ppm) 9.27 (s, 1H), 8.64 (d, J = 4.0 Hz, 1H), 8.32 (d, J = 10.1 Hz, 2H), 7.86 ‐ 7.79 (m, 1H), 7.60 (t, J = 5.6 Hz, 1H), 5.44 (d, J = 3.3 Hz, 1H), 4.85 (d, J = 14.7 Hz, 1H), 4.60 (d, J = 14.9 Hz, 1H), 4.16 (d, J = 10.1 Hz, 1H), 4.05 (d, J = 10.5 Hz, 1H), 3.19 (s, 1H), 2.58 (d, J = 12.4 Hz, 2H), 2.46 ‐ 2.19 (m, 3H), 1.47 (t, J = 3.2 Hz, 6H), 0.95 (d, J = 3.6 Hz, 3H).

[0387] rac-1-(((5S,7R)-3-(5-(2-Hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-8-methylidene-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile Potassium tert-butoxide (106.38 mg, 0.95 mmol) was added to a solution of methyltriphenylphosphonium bromide (337.36 mg, 0.95 mmol) in benzene (3 mL). To this mixture was added dropwise a solution of rac-1-(((5S,7S)-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2,8-dioxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (300 mg, 0.63 mmol) in benzene (3 mL). The reaction mixture was stirred at 80 °C overnight. The reaction mixture was cooled to room temperature, diluted with water (10 mL), and extracted with ethyl acetate (3 x 10 mL). The combined organic layers were washed with brine (30 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under vacuum to give the crude product. The residue was purified by silica gel column chromatography, eluting with methanol in dichloromethane from 0% to 3%, to give rac-1-(((5S,7R)-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-8-methylene-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (100 mg) as a yellow solid. LCMS (ESI-MS) m / z = 473 [M+H] + . 11H NMR (400 MHz, DMSO-d6) δ (ppm) 9.28 (d, J = 1.5 Hz, 1H), 8.64 (d, J = 1.5 Hz, 1H), 8.37 (d, J = 1.4 Hz, 1H), 8.21 (s, 1H), 7.80 (d, J = 8.3 Hz, 1H), 7.57 (dd, J = 8.3, 1.4 Hz, 1H), 5.44 (s, 1H), 5.00 (s, 1H), 4.76 (d, J = 14.6 Hz, 1H), 4.49 (s, 1H), 4.44 (d, J = 14.6 Hz, 1H), 4.03 ‐ 3.92 (m, 2H), 2.97 (dd, J = 15.2, 11.1 Hz, 1H), 2.40 (ddd, J = 24.0, 14.3, 3.4 Hz, 2H), 2.28 (d, J = 13.1 Hz, 1H), 1.86 ‐ 1.74 (m, 2H), 1.50 ‐ 1.45 (m, 6H), 0.90 (s, 3H).

[0388] 1-(((5S,7R)-3-(5-(2-Hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-8-methylidene-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile and 1-(((5R,7S)-3-(5-(2-Hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-8-methylidene-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000216.tif53165rac-1-(((5S,7R)-3-(5-(2-Hydroxypropan-2-yl)pyrazin-2-yl)-8,8-dimethyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (100 mg, 0.21 mmol) was separated by preparative chiral HPLC under the conditions: column: CHIRAL ART Amylose-SA, 2 * 25 cm, 5 μm, mobile phase A: MtBE (0.5% 2M NH3-MeOH)--HPLC, mobile phase B: EtOH--HPLC, flow rate: 20 mL / min, gradient: 20% B to 20% B in 22 minutes; wavelength: 220 / 254 nm; RT1 (min): 8.922, RT2 (min): 16.131, sample solvent: EtOH--HPLC, injection volume: 1.3 mL. The desired fractions were combined and lyophilized to obtain the product.

[0389] The first eluting isomer: 1-(((5S,7R)-3-(5-(2-Hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-8-methylene-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile as a white solid (28.0 mg, purity 99.4%, 100% ee, yield 28.0%). 11H NMR (400 MHz, DMSO-d6) δ(ppm) 9.28 (d, J = 1.5 Hz, 1H), 8.64 (d, J = 1.5 Hz, 1H), 8.37 (d, J = 1.4 Hz, 1H), 8.21 (s, 1H), 7.80 (d, J = 8.3 Hz, 1H), 7.57 (dd, J = 8.3, 1.4 Hz, 1H), 5.44 (s, 1H), 5.00 (s, 1H), 4.76 (d, J = 14.6 Hz, 1H), 4.49 (s, 1H), 4.44 (d, J = 14.6 Hz, 1H), 4.03 ‐ 3.92 (m, 2H), 2.97 (dd, J = 15.2, 11.1 Hz, 1H), 2.40 (ddd, J = 24.0, 14.3, 3.4 Hz, 2H), 2.28 (d, J = 13.1 Hz, 1H), 1.86 ‐ 1.74 (m, 2H), 1.50 ‐ 1.45 (m, 6H), 0.90 (s, 3H). LCMS (ESI-MS) m / z = 473 [M+H] + .

[0390] Second eluting isomer: 1-(((5R,7S)-3-(5-(2-Hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-8-methylidene-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile as a white solid (32.2 mg, purity 99.7%, 99.8% ee, yield 32.2%). 11H NMR (400 MHz, DMSO-d6) δ(ppm) 9.28 (d, J = 1.5 Hz, 1H), 8.64 (d, J = 1.5 Hz, 1H), 8.37 (d, J = 1.4 Hz, 1H), 8.21 (s, 1H), 7.80 (d, J = 8.3 Hz, 1H), 7.57 (dd, J = 8.3, 1.4 Hz, 1H), 5.44 (s, 1H), 5.00 (s, 1H), 4.76 (d, J = 14.6 Hz, 1H), 4.49 (s, 1H), 4.44 (d, J = 14.6 Hz, 1H), 4.03 ‐ 3.92 (m, 2H), 2.97 (dd, J = 15.2, 11.1 Hz, 1H), 2.40 (ddd, J = 24.0, 14.3, 3.4 Hz, 2H), 2.28 (d, J = 13.1 Hz, 1H), 1.86 ‐ 1.74 (m, 2H), 1.50 ‐ 1.45 (m, 6H), 0.90 (s, 3H). LCMS (ESI-MS) m / z = 473 [M+H] + .

[0391] Example 17. Preparation of 1-(((5R,7S)-3-(5-(2-Hydroxypropan-2-yl)pyrazin-2-yl)-7,9,9-trimethyl-2-oxo-1-oxa-3-azaspiro[[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000217.tif58165

[0392] Detailed procedure 3,5,5-Trimethylcyclohex-2-en-1-one In a stirred mixture of ethyl acetoacetate (21 mL, 161.54 mmol) in tert-butanol (170 mL), potassium tert-butoxide (0.94 g, 8.38 mmol) and 4-methylpent-3-en-2-one (15.84 g, 161.54 mmol) were added, and the resulting mixture was stirred at room temperature for 0.5 h. Then, another batch of potassium tert-butoxide (3.63 g, 32.31 mmol) was added, and the resulting mixture was heated to 80 °C and stirred for 16 h. The reaction mixture was cooled to room temperature, filtered, and the filtrate was concentrated to give a crude product. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0% - 15% petroleum ether to give 3,5,5-trimethylcyclohex-2-en-1-one (15 g) as a colorless oil. LCMS (ESI-MS) m / z = 139 [M+H] + .

[0393] rac-3,3,5-Trimethyl-5-(nitromethyl)cyclohexan-1-one In a stirred mixture of 3,5,5-trimethylcyclohex-2-en-1-one (15 g, 108.53 mmol) in methanol (90 mL), nitromethane (6.62 g, 108.53 mmol) and pyrrolidine (7.72 g, 108.53 mmol) were added. The resulting mixture was stirred at room temperature for 16 h and concentrated under vacuum to give a crude product. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0% - 10% petroleum ether to give the desired product rac-3,3,5-trimethyl-5-(nitromethyl)cyclohexan-1-one (7 g) as a yellow oil. LCMS (ESI-MS) m / z = 200 [M+H] + .

[0394] rac-7,7,9-Trimethyl-9-(nitromethyl)-1,4-dioxaspiro[4.5]decane In dichloromethane (40 mL), to a stirred mixture of rac-3,3,5-trimethyl-5-(nitromethyl)cyclohexan-1-one (7 g, 35.13 mmol), ethylene glycol (2.18 g, 35.13 mmol), trimethoxymethane (3.73 g, 35.13 mmol) and methanesulfonic acid (0.51 g, 5.27 mmol) were added. The resulting mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated under vacuum to afford a crude product. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in 0% - 15% petroleum ether to give the desired product rac-7,7,9-trimethyl-9-(nitromethyl)-1,4-dioxaspiro[4.5]decane (5 g) as a yellow oil. LCMS (ESI-MS) m / z = 244 [M+H] + .

[0395] rac-7,9,9-trimethyl-1,4-dioxaspiro[4.5]decan-7-yl)methanamine In ethanol (75 mL) and water (15 mL), to a stirred mixture of rac-7,7,9-trimethyl-9-(nitromethyl)-1,4-dioxaspiro[4.5]decane (5 g, 20.55 mmol), iron (5.74 g, 102.75 mmol) and ammonium chloride (3.30 g, 61.65 mmol) were added. The resulting mixture was heated to 80 °C and stirred for 2 hours. The resulting mixture was cooled to room temperature, filtered, and the filter cake was washed with ethanol (50 mL). The filtrate was diluted with water (50 mL), extracted with dichloromethane (3 × 50 mL), the combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give rac-(7,9,9-trimethyl-1,4-dioxaspiro[4.5]decan-7-yl)methanamine (3 g) as a yellow oil. LCMS (ESI-MS) m / z = 214 [M+H] + .

[0396] rac-4-Nitro-3-(((7,9,9-trimethyl-1,4-dioxaspiro[4.5]decane-7-yl)methyl)amino)benzonitrile In acetonitrile (60 mL), to a stirred mixture of rac-(7,9,9-trimethyl-1,4-dioxaspiro[4.5]decane-7-yl)methanamine (3 g, 14.06 mmol), 3-fluoro-4-nitrobenzonitrile (2.34 g, 14.06 mmol) and potassium carbonate (3.89 g, 28.12 mmol) were added. The resulting mixture was heated to 70 °C and stirred overnight. The reaction mixture was cooled to room temperature, filtered, and the filtrate was concentrated under vacuum to give the crude product. The residue was purified by silica gel column chromatography, eluting with ethyl acetate in petroleum ether containing 0% - 10% ethyl acetate to give the desired product rac-4-nitro-3-(((7,9,9-trimethyl-1,4-dioxaspiro[4.5]decane-7-yl)methyl)amino)benzonitrile (5 g) as an orange solid. LCMS (ESI-MS) m / z = 360 [M+H] + .

[0397] rac-4-Amino-3-(((7,9,9-trimethyl-1,4-dioxaspiro[4.5]decane-7-yl)methyl)amino)benzonitrile In ethanol (60 mL) and water (20 mL), to a stirred mixture of rac-4-nitro-3-(((7,9,9-trimethyl-1,4-dioxaspiro[4.5]decane-7-yl)methyl)amino)benzonitrile (3 g, 8.34 mmol), iron (2.33 g, 41.73 mmol) and ammonium chloride (2.23 g, 41.73 mmol) were added. The resulting mixture was heated to 80 °C and stirred for 2 h. The mixture was cooled to room temperature, filtered, and the filter cake was washed with ethanol (50 mL). The filtrate was diluted with water (50 mL), extracted with dichloromethane (3 × 50 mL), the combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give the desired product rac-4-amino-3-(((7,9,9-trimethyl-1,4-dioxaspiro[4.5]decane-7-yl)methyl)amino)benzonitrile (2.2 g) as a yellow solid. LCMS (ESI-MS) m / z = 330 [M+H] + .

[0398] rac-1-((7,9,9-trimethyl-1,4-dioxaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile In acetonitrile (10 mL), formic acid (0.25 mL, 6.67 mmol) and trimethoxymethane (0.73 mL, 6.67 mmol) were added to a stirred mixture of rac-4-amino-3-(((7,9,9-trimethyl-1,4-dioxaspiro[4.5]decan-7-yl)methyl)amino)benzonitrile (2.2 g, 6.67 mmol). The resulting mixture was heated to 80 °C and stirred for 3 h. The reaction mixture was cooled to room temperature, diluted with water (50 mL) and extracted with dichloromethane (3 x 30 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give the crude product rac-1-((7,9,9-trimethyl-1,4-dioxaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (2 g) as a yellow oil. LCMS (ESI-MS) m / z = 340 [M+H] +

[0399] rac-1-((1,3,3-Trimethyl-5-oxocyclohexyl)methyl)-1H-benzo[d]imidazole-6-carbonitrile In formic acid (6 mL, 159.04 mmol), a mixture of rac-1-((7,9,9-trimethyl-1,4-dioxaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (2 g, 5.89 mmol) was stirred at 80 °C overnight. The mixture was cooled to room temperature and concentrated in vacuo to give the crude product rac-1-((1,3,3-trimethyl-5-oxocyclohexyl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (1.7 g) as a yellow oil. LCMS (ESI-MS) m / z = 296 [M+H] + .

[0400] rac-1-(((3R,5S)-5,7,7-Trimethyl-1-oxaspiro[2.5]octan-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile In dimethyl sulfoxide (20 mL), trimethylsulfoxonium iodide (1.27 g, 5.75 mmol) and potassium tert-butoxide (0.97 g, 8.63 mmol) were added to a stirred mixture of rac-1-((1,3,3-trimethyl-5-oxocyclohexyl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (1.7 g, 5.75 mmol). The resulting mixture was stirred at room temperature for 2 h. The reaction mixture was diluted with water (50 mL) and extracted with dichloromethane (3 x 50 mL). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give the crude product rac-1-(((3R,5S)-5,7,7-trimethyl-1-oxaspiro[2.5]octan-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (1.5 g) as a yellow oil. LCMS (ESI-MS) m / z = 310 [M+H] + .

[0401] rac-1-(((5R,7S)-7,9,9-trimethyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000227.tifA mixture of lithium tert-butoxide (90.56 mg, 1.13 mmol) and ethyl carbamate (1007.80 mg, 11.31 mmol) in 1-methyl-2-pyrrolidinone (6 mL) was stirred at room temperature for 5 minutes, then rac-1-(((3R,5S)-5,7,7-trimethyl-1-oxaspiro[2.5]octan-5-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (700 mg, 2.26 mmol) was added. The resulting mixture was heated to 75 °C and stirred for 16 hours. The reaction mixture was cooled to room temperature, diluted with water (60 mL), and extracted with dichloromethane (3 x 60 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to give the crude product rac-1-(((5R,7S)-7,9,9-trimethyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (500 mg) as a yellow solid. LCMS (ESI-MS) m / z = 353 [M+H] + .

[0402] rac-1-(((5R,7S)-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7,9,9-trimethyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile In dioxane (12 mL), to a stirred mixture of rac-1-(((5R,7S)-7,9,9-trimethyl-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (500 mg, 1.42 mmol), 2-(5-chloropyrazin-2-yl)propan-2-ol (244.88 mg, 1.42 mmol), tribasic potassium phosphate (903.41 mg, 4.25 mmol), methyl[2-(methylamino)ethyl]amine (300.15 mg, 3.41 mmol) and cuprous iodide (270.19 mg, 1.42 mmol) were added under a nitrogen atmosphere. The resulting mixture was heated to 100 °C and stirred for 16 h. The reaction mixture was cooled to room temperature, diluted with water (60 mL) and extracted with ethyl acetate (3 x 60 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo to give the crude product. The residue was purified by silica gel column chromatography, eluting with 0% - 10% methanol in dichloromethane to afford the desired product rac-1-(((5R,7S)-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7,9,9-trimethyl-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (450 mg) as a yellow solid. LCMS (ESI-MS) m / z = 489 [M+H] + .

[0403] 1-(((5S,7R)-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7,9,9-trimethyl-2-oxo-1-oxa-3-azaspiro[4.5 ]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile and 1-(((5R,7S)-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7,9,9-trimethyl-2-oxo-1-oxa-3-azaspiro[4.5]decane-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile The racemic compound of TIFF2025522953000229.tif631651 - (((5R,7S)-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7,9,9-trimethyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (350 mg) was separated by preparative chiral HPLC under the following conditions: column: CHIRALPAK IA, 3 × 25 cm, 5 μm, mobile phase A: hexane (0.5% 2M NH3-MeOH)--HPLC, mobile phase B: EtOH--HPLC, flow rate: 18 mL / min, gradient: 50%B to 50%B in 28 minutes, wavelength: 220 / 254 nm; RT1 (min): 17.68; RT2 (min): 23.745, sample solvent: EtOH--HPLC, injection volume: 0.8 mL. The desired fractions were combined and lyophilized to obtain two products.

[0404] The first eluting isomer: obtained as a white solid, 1-(((5S,7R)-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7,9,9-trimethyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (109.8 mg, purity 99.8%, 100.0% ee. LCMS (ESI-MS) m / z = 489 [M+H] + . 11H NMR (300 MHz, DMSO-d6) δ (ppm) 9.30 (d, J = 1.5 Hz, 1H), 8.62 (d, J = 1.5 Hz, 1H), 8.52 (s, 1H), 8.45 (d, J = 1.5 Hz, 1H), 7.86 (d, J = 8.4 Hz, 1H), 7.62 (dd, J = 8.4, 1.5 Hz, 1H), 4.72 (d, J = 14.5 Hz, 1H), 4.35 (d, J = 14.4 Hz, 2H), 4.05 ‐ 3.91 (m, 2H), 2.17 (d, J = 14.5 Hz, 1H), 1.97 (d, J = 14.4 Hz, 1H), 1.79 (d, J = 14.2 Hz, 1H), 1.67 (d, J = 14.4 Hz, 1H), 1.56 (d, J = 14.9 Hz, 1H), 1.47 (d, J = 3.4 Hz, 6H), 1.35 (d, J = 14.3 Hz, 1H), 1.27 (s, 3H), 1.04 (s, 3H), 0.80 (s, 3H).

[0405] Second eluting isomer: 1-(((5R,7S)-3-(5-(2-Hydroxypropan-2-yl)pyrazin-2-yl)-7,9,9-trimethyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile as a white solid (109.3 mg, purity 96.5%, 98.9% ee). LCMS (ESI-MS) m / z = 489 [M+H] + . 11H NMR (300 MHz, DMSO-d6) δ (ppm) 9.30 (d, J = 1.5 Hz, 1H), 8.62 (d, J = 1.5 Hz, 1H), 8.55 (s, 1H), 8.46 (d, J = 1.5 Hz, 1H), 7.86 (d, J = 8.4 Hz, 1H), 7.63 (dd, J = 8.4, 1.5 Hz, 1H), 4.73 (d, J = 14.4 Hz, 1H), 4.35 (d, J = 14.5 Hz, 1H), 4.03 ‐ 3.95 (m, 2H), 2.17 (d, J = 14.9 Hz, 1H), 1.97 (d, J = 14.4 Hz, 1H), 1.79 (d, J = 14.2 Hz, 1H), 1.67 (d, J = 14.3 Hz, 1H), 1.56 (d, J = 14.8 Hz, 1H), 1.47 (d, J = 3.5 Hz, 6H), 1.35 (d, J = 14.2 Hz, 1H), 1.27 (s, 3H), 1.04 (s, 3H), 0.80 (s, 3H).

[0406] Example 18. Preparation of 5-Fluoro-1-(((5S,7S,8R)-8-fluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000230.tif53165

[0407] Reaction Scheme TIFF2025522953000231.tif53165

[0408] Detailed Procedure rac-5-Fluoro-1-(((5S,7S,8R)-8-fluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile Copper(I) iodide (53.3 mg, 0.28 mmol) was added under nitrogen to a solution of rac-5-fluoro-1-(((5S,7S,8R)-8-fluoro-7-methoxy-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (100 mg, 0.28 mmol), 2-(5-chloropyridin-2-yl)propan-2-ol (48.3 mg, 0.28 mmol), N 1 ,N 2 -dimethylethane-1,2-diamine (49.4 mg, 0.56 mmol) and tripotassium phosphate (119 mg, 0.56 mmol) in 1,4-dioxane (2 mL). The resulting mixture was heated to 100 °C and stirred overnight. After cooling to room temperature, the resulting mixture was filtered and the filtrate was concentrated under vacuum. The residue was purified by silica gel column chromatography, eluting with methanol in dichloromethane from 0 to 2%. The fractions were combined and concentrated under vacuum to give the desired product rac-5-fluoro-1-(((5S,7S,8R)-8-fluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (100 mg) as a pale yellow solid. 11H NMR (400 MHz, DMSO-d6) δ 9.18 (d, J = 1.5 Hz, 1H), 8.58 (d, J = 1.5 Hz, 1H), 8.50 (s, 1H), 8.33 (d, J = 5.6 Hz, 1H), 7.81 (d, J = 10.1 Hz, 1H), 5.42 (s, 1H), 4.69 - 4.51 (m, 1H), 4.31 (d, J = 2.7 Hz, 2H), 3.84 (s, 2H), 2.13 - 1.96 (m, 2H), 1.95 - 1.87 (m, 2H), 1.83 - 1.66 (m, 2H), 1.44 (d, J = 2.0 Hz, 6H), 1.18 (s, 3H). LCMS (ESI-MS) m / z = 497 [M+H] + .

[0409] 5-Fluoro-1-(((5S,7S,8R)-8-fluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile and 5-fluoro-1-(((5R,7R,8S)-8-fluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000233.tif 5-fluoro-1-(((5S,7S,8R)-8-fluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (100 mg, 0.20 mmol) was separated by preparative chiral HPLC under the conditions: CHIRALPAK IE, 2 * 25 cm, 5 μm, mobile phase A: MTBE (0.5% 2M NH3-MeOH)--HPLC, mobile phase B: EtOH--HPLC, flow rate: 20 mL / min, gradient: isocratic 20, wavelength: 254 / 220 nm, RT1 (min): 8.262, RT2 (min): 10.453, sample solvent: EtOH--HPLC, injection volume: 1.0 mL. The desired fractions were combined and lyophilized to give the product.

[0410] The first eluting isomer: 5-fluoro-1-(((5S,7S,8R)-8-fluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile as a white solid (20.2 mg, purity 98.3%, 100% ee, yield 20.2%). 1 H NMR (400 MHz, DMSO-d6) δ 9.18 (d, J = 1.5 Hz, 1H), 8.58 (d, J = 1.5 Hz, 1H), 8.50 (s, 1H), 8.33 (d, J = 5.6 Hz, 1H), 7.81 (d, J = 10.1 Hz, 1H), 5.42 (s, 1H), 4.69 - 4.51 (m, 1H), 4.31 (d, J = 2.7 Hz, 2H), 3.84 (s, 2H), 2.13 - 1.96 (m, 2H), 1.95 - 1.87 (m, 2H), 1.83 - 1.66 (m, 2H), 1.44 (d, J = 2.0 Hz, 6H), 1.18 (s, 3H). LCMS (ESI-MS) m / z = 497 [M + H] + .

[0411] Second eluting isomer: 5-Fluoro-1-(((5R,7R,8S)-8-fluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile as a white solid (20.0 mg, purity 96.4%, 99.5% ee, yield 20.0%). 1 H NMR (400 MHz, DMSO-d6) δ 9.18 (d, J = 1.5 Hz, 1H), 8.58 (d, J = 1.6 Hz, 1H), 8.50 (s, 1H), 8.33 (d, J = 5.6 Hz, 1H), 7.81 (d, J = 10.1 Hz, 1H), 5.41 (s, 1H), 4.69 - 4.51 (m, 1H), 4.31 (d, J = 2.7 Hz, 2H), 3.84 (s, 2H), 2.13 - 1.99 (m, 2H), 1.97 - 1.88 (m, 2H), 1.83 - 1.67 (m, 2H), 1.44 (d, J = 2.0 Hz, 6H), 1.17 (s, 3H). LCMS (ESI-MS) m / z = 497 [M+H] + .

[0412] Example 19. Preparation of 4-chloro-1-(((5S,7S,8R)-8-fluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000234.tif68165

[0413] Reaction scheme TIFF2025522953000235.tif53165

[0414] Detailed procedure rac-4-chloro-1-(((5S,7S,8R)-8-fluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000236.tif53165Copper(I) iodide (51.4 mg, 0.27 mmol) was added under nitrogen to a solution of rac-4-chloro-1-(((5S,7S,8R)-8-fluoro-7-methoxy-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (100 mg, 0.27 mmol), 2-(5-chloropyridin-2-yl)propan-2-ol (46.6 mg, 0.27 mmol), N 1 ,N 2 -dimethylethane-1,2-diamine (47.6 mg, 0.54 mmol) and tripotassium phosphate (114.6 mg, 0.54 mmol) in 1,4-dioxane (2 mL). The resulting mixture was heated to 100 °C and stirred overnight. After cooling to room temperature, the resulting mixture was filtered and the filtrate was concentrated under vacuum. The residue was purified by silica gel column chromatography, eluting with methanol in dichloromethane from 0 to 2%. Fractions having the desired mass signal were combined and concentrated under vacuum to give the desired product rac-4-chloro-1-((5S,7S,8R)-8-fluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (100 mg) as a pale yellow solid. 11H NMR (400 MHz, DMSO-d6) δ 9.17 (d, J = 1.5 Hz, 1H), 8.58 (d, J = 1.5 Hz, 1H), 8.53 (s, 1H), 8.29 (d, J = 1.3 Hz, 1H), 7.82 (d, J = 1.3 Hz, 1H), 5.41 (s, 1H), 4.68 - 4.50 (m, 1H), 4.40 - 4.27 (m, 2H), 3.82 (d, J = 1.4 Hz, 2H), 2.12 - 1.98 (m, 2H), 1.96 - 1.88 (m, 2H), 1.82 - 1.67 (m, 2H), 1.44 (d, J = 2.3 Hz, 6H), 1.19 (s, 3H). LCMS (ESI-MS) m / z = 513 [M+H] + .

[0415] 4-chloro-1-(((5S,7S,8R)-8-fluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile and 4-chloro-1-(((5R,7R,8S)-8-fluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000237.tif53165rac-4-chloro-1-(((5S,7S,8R)-8-fluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile (100 mg, 0.19 mmol) was separated by preparative chiral HPLC under the conditions: CHIRALPAK IE, 2 * 25 cm, 5 μm, mobile phase A: MtBE (0.5% 2M NH3-MeOH)--HPLC, mobile phase B: EtOH--HPLC, flow rate: 19 mL / min, gradient: isocratic 30, wavelength: 254 / 220 nm, RT1 (min): 8.246, RT2 (min): 11.53, sample solvent: EtOH--HPLC, injection volume: 0.8 mL. The desired fractions were combined and lyophilized to obtain the product.

[0416] The first eluting isomer: 4-chloro-1-(((5R,7R,8S)-8-fluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile as a white solid (42.0 mg, purity 99.9%, 100% ee). 1 H NMR (400 MHz, DMSO-d6) δ 9.17 (d, J = 1.5 Hz, 1H), 8.58 (d, J = 1.5 Hz, 1H), 8.53 (s, 1H), 8.29 (d, J = 1.3 Hz, 1H), 7.82 (d, J = 1.3 Hz, 1H), 5.41 (s, 1H), 4.68 - 4.50 (m, 1H), 4.40 - 4.27 (m, 2H), 3.82 (d, J = 1.4 Hz, 2H), 2.12 - 1.98 (m, 2H), 1.96 - 1.88 (m, 2H), 1.82 - 1.67 (m, 2H), 1.44 (d, J = 2.3 Hz, 6H), 1.19 (s, 3H). LCMS (ESI-MS) m / z = 513 [M+H] + .

[0417] Second eluting isomer: 4-chloro-1-(((5S,7S,8R)-8-fluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile as a white solid (45.3 mg, purity 98.8%, 99.7% ee). 1 H NMR (400 MHz, DMSO-d6) δ 9.17 (d, J = 1.5 Hz, 1H), 8.58 (d, J = 1.5 Hz, 1H), 8.53 (s, 1H), 8.29 (d, J = 1.3 Hz, 1H), 7.82 (d, J = 1.3 Hz, 1H), 5.41 (s, 1H), 4.68 - 4.50 (m, 1H), 4.40 - 4.27 (m, 2H), 3.82 (d, J = 1.4 Hz, 2H), 2.12 - 1.98 (m, 2H), 1.96 - 1.88 (m, 2H), 1.82 - 1.67 (m, 2H), 1.44 (d, J = 2.3 Hz, 6H), 1.19 (s, 3H). LCMS (ESI-MS) m / z = 513 [M+H] + .

[0418] Example 20. Preparation of 1-(((5S,7S,8S)-8-fluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000238.tif63165The title compound was prepared using a procedure similar to the procedure described in this specification (see Scheme 7).

[0419] Example 21. Preparation and Characterization of a Crystal Form of 1-(((5S,7S,8R)-8-Fluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile TIFF2025522953000239.tif63165249.15 mg of 1-(((5S,7S,8R)-8-fluoro-3-(5-(2-hydroxypropan-2-yl)pyrazin-2-yl)-7-methyl-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole-6-carbonitrile was weighed and placed in a 40 mL glass vial, 10 mL of anhydrous MeOH was added to the vial, and the mixture was stirred (at 700 rpm) at 25 °C for 3 days. The suspension was dried in a vacuum oven at 30 °C for 3 hours and subsequently characterized by XRPD (Figure 1).

[0420] The diffractogram was obtained at room temperature using a Bruker D8 Advance powder diffractometer with copper Kα radiation (40 kV / 40 mA), a range of 3 - 40 degrees 2θ with 0.02 step size, a step time of 0.12 seconds, and a detector with a window of 10 degrees. The selected peaks are shown in the following table. TIFF2025522953000240.tif61165

[0421] Example 22. Preparation of Additional Compounds The following compounds can be prepared using known procedures and intermediates or procedures and intermediates similar to those described in the above examples. TIFF2025522953000241.tif126165

[0422] Example 23. TRPV4 Calcium Assay Method Transfection Human embryonic kidney 293T (HEK293T / 293T) cells are cultured in complete medium (Dulbecco's Modified Eagle Medium (DMEM) containing 10% fetal bovine serum (FBS) and 1x penicillin-streptomycin) in a T-75 tissue culture flask. At 90% confluence, cells are detached using TrypLE™ Express cell dissociation buffer. The TripLE™ Express buffer is neutralized by adding excess medium. Cells are gently and thoroughly triturated and counted using trypan blue in a Countess 3 automated cell counter. Cells are transferred to a 50 mL conical tube and spun down at 1000 revolutions per minute (RPM) for 5 minutes in a benchtop centrifuge. The medium containing the detachment enzyme is aspirated from the pellet and subsequently resuspended in complete DMEM to a desired concentration of 2×10 6 cells / mL.

[0423] The transfection preparation is scaled using a combination of 6 μg of TRPV4 wild-type or mutant plasmid DNA, 36 μL of Fugene6 transfection reagent, and 1000 μL of OptiMEM medium per 2×10 6 cells. The transfection mix is left to incubate at room temperature for 20 minutes. The total seeding mixture for one transfection (1000 μL of 2×10 6Mix the cells, 1000 μL of the aforementioned transfection mix, and 1000 μL of complete DMEM) in a 15 mL conical tube. Mix the cells and the transfection plasmid well and dispense into a reservoir. Plate this at 25 μL per well onto a black 384-well, black well, clear bottom, tissue culture-treated plate. This yields approximately 16,000 cells per well. The plasmid DNA constructs were designed by Acctio Biosciences in collaboration with Genscript and generated by Genscript for all hTRPV4 wild-type and hTRPV4 P / LP (ClinVar) mutations (G78W, P82S, T89I, P97R, R186Q, R232C, R232P, R237G, R237Q, R237L, R269C, R269H, R269S, G270V, R271P, F273L, K276E, E278K, G280S, Y281S, R315G, R315W, R316C, R316H, R316L, R316S, I331F, D333G, S403T, K403T, K407E, S542P, S542Y, L543R, Y591N, R594C, R594H, R594S, G600E, Y602C, R616Q, F617L, L618P, V620I, A716S, A716T, L735R, T740I, R775K, W785R, E797D, E797K, P799A, P799R, P799S, P799L, G800D, K801E, P827H, F471del, N797del). Plate the remaining cells in the same manner onto a 384-well, white well, clear bottom, tissue culture-treated plate for determination of expression and relative cell number. Incubate the plates overnight in a 37 °C incubator.

[0424] Determination of relative cell number The relative cell number was determined using the Promega (trademark)-manufactured Cell-Titer Fluor (CTF) assay kit. The reagents were prepared according to the manufacturer's instructions. An equal volume of CTF reagent was added to each well. The cell plate was returned to the incubator at 37 °C for 30 minutes. After 30 minutes, the plate was read on a BMG CLARIOstar using an EX / EM setting: excitation 380 - 400 nm / emission 505 nm.

[0425] Determination of expression Nano-Glo (registered trademark) Lytic reagent was prepared according to the manufacturer's instructions. This assay was compatible with the aforementioned CTF assay. Using the same plate as the cells obtained from the CTF assay, an equal volume of HiBit lysis reagent was added to each well. After leaving the reagent and cells at room temperature for 10 minutes, luminescence was read on a BMG CLARIOstar.

[0426] Preparation of compounds Agonist mode The TRPV4 agonist was made 10 mM in DMSO. The concentration of DMSO was kept constant throughout the dilution of any compound. The DMSO negative control and concentration - response curves were spotted in nanoliters at 6-fold the test concentration via a PICO8 automatic dispenser onto a 384w clear polypropylene plate containing 1xBSS + 20 mM HEPES buffer.

[0427] Antagonist mode The compound was made 10 mM in DMSO. The concentration of DMSO was kept constant throughout the dilution of any compound. The DMSO negative control and concentration - response curves were spotted in nanoliters at 5-fold the test concentration via a PICO8 automatic dispenser onto a 384w clear polypropylene plate containing 1xBSS + 20 mM HEPES buffer. The predetermined concentration corresponding to the EC 80 of the TRPV4 agonist for hTRPV4 was prepared 6-fold in 1xBSS + 20 mM HEPES and loaded into another 384-well v-bottom polypropylene plate.

[0428] Calcium Flux Assay Approximately 18 hours after transfection, the cell plates were removed from the incubator and an equal volume (25 uL) of Calcium 6 Dye was gently added. Calcium 6 Dye containing probenecid (final 5 mM) was prepared according to the manufacturer's instructions. The cell plates were returned to the incubator at 37 °C for 1 hour.

[0429] The FLIPR Penta was programmed to add the compound using the following settings. Pipetter Head = 384 Read Mode 1 Excitation Wavelength = 470 - 495 Read Mode 1 Emission Wavelength = 515 - 575 Read Mode 1 Exposure Time = 0.1000 Read Mode 1 Gain = 6.50 Read Mode 1 Excitation Intensity = 20 Volume of Addition = 12.5 μL Height = 40 μL Speed = 15 μL / sec

[0430] The raw data was exported and analyzed for the maximum signal (RFU) after agonist addition - the average baseline (RFU) for the first 15 seconds per well. The data was plotted in GraphPad Prism to determine the EC 50 or IC 50 of the molecule. Data for representative compounds is provided in the following table.

[0431] Example 24. CYP3A4 Incubation Assay The compound (final concentration 1 mM) was incubated in 100 mM phosphate buffer containing 100 pmol / mL of recombinant human CYP3A4 enzyme for 0, 5, 10, 15, and 25 minutes. Testosterone was used as a positive control. The reaction was initiated by the addition of NADPH (final concentration 1 mM) and carried out at 37 °C. The reaction was stopped by the addition of 4 volumes of cold acetonitrile (having 100 nM alprazolam, 200 nM labetalol, 200 nM imipramine, and 2 μM ketoprofen as internal standards). The samples were centrifuged at 3,220 g for 40 minutes to precipitate the protein. 70 μL of the supernatant was transferred to an assay plate containing 110 μL of acetonitrile for LC-MS / MS analysis.

[0432] Data for representative compounds of formula (I) obtained from the assays of Examples 23 and 24 are shown in the following table. TIFF2025522953000242.tif169165TIFF2025522953000243.tif173165TIFF2025522953000244.tif168165TIFF2025522953000245.tif197165TIFF2025522953000246.tif165165TIFF2025522953000247.tif221165

[0433] Example 25. Incubation with CYP3A4 The following table shows the percentage remaining for the examples shown after incubation with recombinant human CYP3A4 for 25 minutes under the same conditions as described for Example 23. TIFF2025522953000248.tif42165

[0434] Example 26 Selectivity for the compound of Example 5 against TRP The compound of Example 5 was tested for selectivity against a panel of human TRP family ion channels in the TRP family FLIPR assay as described below. 1. TRPC3 channel (human TRPC3 variant 1 gene expressed in HEK293 cells). 2. TRPC4 channel (human TRPC4 gene expressed in HEK293 cells). 3. TRPC5 channel (human TRPC5 gene expressed in HEK293 cells). 4. TRPC6 channel (human TRPC6 gene expressed in HEK293 cells). 5. TRPM2 channel (human TRPM2 gene transiently expressed in HEK293 cells). 6. TRPM3 channel (human TRPM3 gene transiently expressed in HEK293 cells). 7. TRPM4 channel (human TRPM4 gene expressed in CHO cells). 8. TRPM5 channel (human TRPM5 gene expressed in CHO cells). 9. TRPA1 channel (human TRPA1 gene expressed in CHO cells). 10. TRPV1 channel (human TRPV1 gene expressed in HEK293 cells). 11. TRPV2 channel (human TRPV2 gene transiently expressed in HEK293 cells). 12. TRPV3 channel (human TRPV3 gene transiently expressed in HEK293 cells). 13. TRPV4 channel (human TRPV4 gene expressed in CHO cells). 14. TRPV5 channel (human TRPV5 gene transiently expressed in HEK293 cells). 15. TRPV6 channel (human TRPV6 gene expressed in HEK293 cells).

[0435] Data acquisition was performed via the FLIPR control software attached to the FLIPR system (MDS-AT), and the data was analyzed using Microsoft Excel (Microsoft Corporation, REDmond, WA) or GraphPad Prism. The concentration-response data was fitted to the Hill equation in the following form. TIFF2025522953000249.tif27165(where Base is the reaction in the low - concentration test substance, Max is the maximum reaction at high concentration, xhalf is the EC 50 or IC 50 which is the test substance that brings about half - maximal activation or inhibition, and rate is the Hill coefficient.) Assuming a simple binding model, fitting was performed by non - linear least - squares method. If necessary, the fitting was weighted by the standard deviation. No assumptions were made about the fit parameters. The fit parameters were determined by the algorithm.

[0436] Results and Summary The effects of each test substance were evaluated at eight (8) concentrations: 0.003, 0.01, 0.03, 0.1, 0.3, 1, 3, and 10 μM (n ≤ 4 wells / concentration). The results of the test substances were shown as % of E MAX (in the case of an agonist) and % of the control (in the case of an antagonist). The results of the positive control confirmed the sensitivity of the test system to the activation and inhibition of the ion channel.

[0437] Agonist / antagonist effects of test substances on the TRPC3 channel In the agonist mode, the reference agonist GSK1702934A was used as an internal control (0 - 30 μM). The effect of the test substance was calculated as the % of the E MAX response induced by the reference agonist GSK1702934A (30 μM).

[0438] In the antagonist mode, the channel was activated with a positive control agonist (15 μM GSK1702934A). The inhibition of the signal by the test substance was calculated compared to the signal produced by the agonist alone. The reference antagonist verapamil was used as an internal control.

[0439] Agonist / antagonist effects of test substances on the TRPC4 and TRPC5 channels In the agonist mode, (-)-englerin A, which is the reference agonist, was used as an internal control (0 - 30 μM). The effect of the test substance was calculated as the percentage of the EMAX response induced by (-)-englerin A (30 μM), which is the reference agonist.

[0440] In the antagonist mode, the channel was activated with the positive control agonist, (0.5 μM (-)-englerin A). The inhibition of the signal by the test substance was calculated relative to the signal generated by the agonist alone. Verapamil, which is the reference antagonist, was used as an internal control.

[0441] Agonist / antagonist effects of test substances on the TRPC6 channel In the agonist mode, Na-ATP, which is the reference agonist, was used as an internal control (0 - 100 μM). The effect of the test substance was calculated as the percentage of the EMAX response induced by Na-ATP (100 μM), which is the reference agonist.

[0442] In the antagonist mode, the channel was activated with the positive control agonist (10 μM Na-ATP). The inhibition of the signal by the test substance was calculated relative to the signal generated by the agonist alone. Verapamil, which is the reference antagonist, was used as an internal control.

[0443] Agonist / antagonist effects of test substances on the TRPM2 channel In the agonist mode, H2O2, which is the reference agonist, was used as an internal control (0 - 1000 μM). The effect of the test substance was calculated as the percentage of the E MAX response induced by H2O2 (333 μM), which is the reference agonist.

[0444] In the antagonist mode, the channel was activated with the positive control agonist (300 μM H2O2). The inhibition of the signal by the test substance was calculated relative to the signal generated by the agonist alone. Flufenamic acid (0 - 1000 μM), which is the reference antagonist, was used as an internal control.

[0445] Agonist / antagonist effects of test substances on the TRPM3 channel In the agonist mode, pregnenolone, a reference agonist, was used as an internal control (0 - 100 μM). The effect of the test substance was calculated as the % of the E MAX response induced by pregnenolone (100 μM), a reference agonist.

[0446] In the antagonist mode, the channel was activated with a positive control agonist (pregnenolone 30 μM). The inhibition of the signal by the test substance was calculated compared to the signal generated by the agonist alone. Mefenamic acid, a reference antagonist (0 - 200 μM), was used as an internal control.

[0447] Agonist / antagonist effects of test substances on the TRPM4 channel In the agonist mode, A23187, a reference agonist, was used as an internal control (0 - 30 μM). The effect of the test substance was calculated as the % of the E MAX response induced by A23187 3.3 μM, a reference agonist.

[0448] In the antagonist mode, the channel was activated with a positive control agonist (A23l87 10 μM). The inhibition of the signal by the test substance was calculated compared to the signal generated by the agonist alone. Flufenamic acid, a reference antagonist (0 - 200 μM), was used as an internal control.

[0449] Agonist / antagonist effects of test substances on the TRPM5 channel In the agonist mode, Mg - ATP, a reference agonist, was used as an internal control (0 - 100 μM). The effect of the test substance was calculated as the % of the E MAX response induced by Mg - ATP 33.3 μM, a reference agonist.

[0450] In the antagonist mode, the channel was activated with a positive control agonist (Mg-ATP 75 μM). The inhibition of the signal by the test substance was calculated relative to the signal generated by the agonist alone. Triphenylphosphine oxide (TPPO, 0 - 300 μM), a reference antagonist, was used as an internal control.

[0451] Agonist / antagonist effects of test substances on the TRPA1 channel In the agonist mode, allyl isothiocyanate, a reference agonist, was used as an internal control (0 - 600 μM). The effect of the test substance was calculated as the % of the E MAX response induced by 600 μM allyl isothiocyanate, a reference agonist.

[0452] In the antagonist mode, the channel was activated with a positive control agonist (allyl isothiocyanate 80 μM). The inhibition of the signal by the test substance was calculated relative to the signal generated by the agonist alone. Ruthenium red (0 - 30 μM), a reference antagonist, was used as an internal control.

[0453] Agonist / antagonist effects of test substances on the TRPV1 channel In the agonist mode, capsaicin, a reference agonist, was used as an internal control (0 - 1 μM). The effect of the test substance was calculated as the % of the EMAX response induced by 1 μM capsaicin, a reference agonist.

[0454] In the antagonist mode, the channel was activated with a positive control agonist (capsaicin 0.1 μM). The inhibition of the signal by the test substance was calculated relative to the signal generated by the agonist alone. Ruthenium red (0 - 10 μM), a reference antagonist, was used as an internal control.

[0455] Agonist / antagonist effects of test substances on the TRPV2 channel In the agonist mode, cannabidiol, a reference agonist, was used as an internal control (0 - 300 μM). The effect of the test substance was calculated as the % of the E MAXCalculated as % of the reaction.

[0456] In the antagonist mode, the channel was activated with a positive control agonist (capsaicin 100 μM). The inhibition of the signal by the test substance was calculated relative to the signal generated by the agonist alone. Ruthenium red (0 - 30 μM), a reference antagonist, was used as an internal control.

[0457] Agonist / antagonist effects of test substances on TRPV3 channels In the agonist mode, 2 - aminoethoxydiphenyl borate (2 - APB), a reference agonist, was used as an internal control (0 - 300 μM). The effect of the test substance was calculated as % of the EMAX reaction induced by 300 μM of cannabidiol, a reference agonist. MAX Calculated as % of the reaction.

[0458] In the antagonist mode, the channel was activated with a positive control agonist (2 - APB 50 μM). The inhibition of the signal by the test substance was calculated relative to the signal generated by the agonist alone. Ruthenium red (0 - 30 μM), a reference antagonist, was used as an internal control.

[0459] Agonist / antagonist effects of test substances on TRPV4 channels In the agonist mode, GSK1016790A, a reference agonist, was used as an internal control (0 - 3 μM). The effect of the test substance was calculated as % of the EMAX reaction induced by 3 μM of GSK1016790A.

[0460] In the antagonist mode, the channel was activated with a positive control agonist GSK1016790A (0.3 μM). The inhibition of the signal by the test substance was calculated relative to the signal generated by the agonist alone. Ruthenium red (0 - 10 μM), a reference antagonist, was used as an internal control.

[0461] Antagonist effects of test substances on TRPV5 channels In the antagonist mode, the channel was activated by 60 μM cadmium ions. The inhibition of the signal by the test substance was calculated relative to the signal generated in the control. 2-APB (0 - 300 μM), a reference antagonist, was used as an internal control.

[0462] Antagonistic effect of the test substance on the TRPV6 channel In the antagonist mode, the channel was activated by 0.9 μM calcium ions. The inhibition of the signal by the test substance was calculated relative to the signal generated in the control. 2-APB (0 - 300 μM), a reference antagonist, was used as an internal control.

[0463] The data of the compound of Example 5 are shown in the following table. TIFF2025522953000250.tif111165

[0464] Example 27. Representative pharmaceutical dosage forms containing a compound of formula I ("Compound X") for therapeutic or prophylactic use in humans are exemplified below.

[0465] (i) Tablets 1 mg / tablet Compound X □ 100.0 Lactose 77.5 Povidone 15.0 Croscarmellose sodium 12.0 Microcrystalline cellulose 92.5 Magnesium stearate 3.0 300 (ii) Tablets 2 mg / tablet Compound X □ 20.0 Microcrystalline cellulose 410.0 Starch 50.0 Sodium starch glycolate 15.0 Magnesium stearate 5.0 500.0 (iii) Capsules mg / capsule Compound X □ 10.0 Colloidal silicon dioxide 1.5 Lactose 465.5 α-Starch 120.0 Magnesium stearate 3.0 600.0 (iv) Injection 1 (1 mg / ml) mg / ml Compound X (free acid form) 1.0 Disodium hydrogen phosphate 12.0 Sodium dihydrogen phosphate 0.7 Sodium chloride 4.5 1.0 N Sodium hydroxide solution (pH adjustment to 7.0 - 7.5) qs Water for injection q.s. ad 1 mL (v) Injection 2 (10 mg / ml) mg / ml Compound X (free acid form) 10.0 Sodium dihydrogen phosphate 0.3 Disodium hydrogen phosphate 1.1 Polyethylene glycol 400 200.0 1.0 N Sodium hydroxide solution (pH adjustment to 7.0 - 7.5) q.s. Water for injection q.s. ad 1 mL (vi) Aerosol mg / can Compound X□ 20.0 Oleic acid 10.0 Trichloromonofluoromethane 5,000.0 Dichlorodifluoromethane 10,000.0 Dichlorotetrafluoroethane 5,000.0

[0466] The above preparations can be obtained using conventional procedures well known in the pharmaceutical art.

[0467] All publications, patents, and patent documents are hereby incorporated by reference as if each were individually incorporated by reference. The present disclosure has been described with reference to various specific and preferred embodiments and techniques. However, it should be understood that many variations and modifications can be made while remaining within the spirit and scope of the content of the present disclosure.

Claims

1. Formula I: a compound or a salt thereof, wherein L １ is CR ａ R ｂ and L ２ is CR ｃ R ｄ and R ａ is H, halo, cyano, (C １ -C ５ ), alkyl, (C １ -C ５ ), alkoxy, (C ３ -C ５ ), cycloalkyloxy, or (C ３ -C ５ ), cycloalkyl, and any (C １ -C ５ ), alkyl, (C １ -C ５ ), alkoxy, (C ３ -C ５ ), cycloalkyloxy, and (C ３ -C ５ ), cycloalkyl may be substituted with one or more fluoros, R ｂ is H, halo, cyano, (C １ -C ５ ), alkyl, (C １ -C ５ ), alkoxy, or (C ３ -C ５ ), cycloalkyl, and any (C １ -C ５ ), alkyl, (C １ -C ５ ), alkoxy, and (C ３ -C ５ ), cycloalkyl may be substituted with one or more fluoros, or R ａ and R ｂ together form oxo (=O) methylene (=CH ２ ), or R ａ and R ｂ together with the atoms to which they are attached form spiro(C ３ -C ５ ), cycloalkyl, R ｃ is H, halo, cyano, (C １ -C ５ )-alkyl, (C １ -C ５ )-alkoxy, (C ３ -C ５ )-cyclopropyloxy, or (C ３ -C ５ )-cycloalkyl, and any (C １ -C ５ )-alkyl, (C １ -C ５ )-alkoxy, (C ３ -C ５ )-cyclopropyloxy, and (C ３ -C ５ )-cycloalkyl may be substituted with one or more fluoros, R ｄ is H, halo, cyano, (C １ -C ５ )-alkyl, (C １ -C ５ )-alkoxy, or (C ３ -C ５ )-cycloalkyl, and any (C １ -C ５ )-alkyl, (C １ -C ５ )-alkoxy, and (C ３ -C ５ )-cycloalkyl may be substituted with one or more fluoros, or R ｃ and R １ together with the atom to which they are attached form a fused cyclopropyl ring, or R ｃ and R ｄ together are oxo (=O) or methylene (=CH ２ ), or R ｃ and R ｄ together with the atom to which they are attached form spiro(C ３ -C ５ )-cycloalkyl, R ａ 、R ｂ 、R ｃ 、and R ｄ at least one of which is other than H, R １ is H, hydroxy, cyano, halo, methoxycarbonyl, cyclopropyl, (C １ -C ３ )alkyl, or (C １ -C ３ )alkoxy, and any cyclopropyl, (C １ -C ３ )alkyl, and (C １ -C ３ )alkoxy may be substituted with one or more groups independently selected from hydroxy, (C １ -C ３ )alkoxy, benzyloxy, cyano, and fluoro, or R １ and R ｃ together with the atom to which they are attached form a fused cyclopropyl ring, Each R ２ is independently selected from the group consisting of halo, cyano, hydroxy, ethynyl, (C １ -C ３ )alkylsulfonyl, cyclopropyl, (C １ -C ３ )alkyl, and (C １ -C ３ )alkoxy, and any cyclopropyl, (C １ -C ３ )alkyl, and (C １ -C ３ )alkoxy may be substituted with one or more fluoros, R ４ is (C １ -C ７ ) alkyl, (C ３ -C ７ ) cycloalkyl, phenyl, or 6-membered heteroaryl, and any R ４ may be substituted with one or more R ｘ ; Each R ｘ is independently selected from the group consisting of hydroxy, halo, cyano, (C １ -C ５ ), alkyl, (C １ -C ５ ), alkoxy, (C ３ -C ５ ), cycloalkyloxy, and (C ３ -C ７ ), cycloalkyl, and any (C １ -C ５ ), alkyl, (C １ -C ５ ), alkoxy, (C ３ -C ５ ), cycloalkyloxy, and (C ３ -C ５ ), cycloalkyl may be substituted with one or more groups independently selected from halo, cyano, (C １ -C ５ ), alkyl, and hydroxy, and A is 0, 1, or 2, said compound or a salt thereof, or Formula I: a compound or a salt thereof, wherein L １ is CR ａ R ｂ and L ２ is CR ｃ R ｄ and R ａ is H, halo, cyano, (C １ -C ５ )-alkyl, (C １ -C ５ )-alkoxy, (C ３ -C ５ )-cycloalkyloxy, or (C ３ -C ５ )-cycloalkyl, and any (C １ -C ５ )-alkyl, (C １ -C ５ )-alkoxy, (C ３ -C ５ )-cycloalkyloxy, and (C ３ -C ５ )-cycloalkyl may be substituted with one or more fluoros, R ｂ is H, halo, cyano, (C １ -C ５ )-alkyl, (C １ -C ５ )-alkoxy, or (C ３ -C ５ )-cycloalkyl, and any (C １ -C ５ )-alkyl, (C １ -C ５ )-alkoxy, and (C ３ -C ５ )-cycloalkyl may be substituted with one or more fluoros, or R ａ and R ｂ together are oxo (=O) methylene (=CH ２ ), or R ａ and R ｂ together with the atoms to which they are attached form spiro(C ３ -C ５ )-cycloalkyl, R ｃ is H, halo, cyano, (C １ -C ５ )-alkyl, (C １ -C ５ )-alkoxy, (C ３ -C ５ )-cycloalkyloxy, or (C ３ -C ５ )-cycloalkyl, and any (C １ -C ５ )-alkyl, (C １ -C ５ )-alkoxy, (C ３ -C ５ )-cycloalkyloxy, and (C ３ -C ５ )-cycloalkyl may be substituted with one or more fluoros, R ｄ is H, halo, cyano, (C １ -C ５ )-alkyl, (C １ -C ５ )-alkoxy, or (C ３ -C ５ )-cycloalkyl, and any (C １ -C ５ )-alkyl, (C １ -C ５ )-alkoxy, and (C ３ -C ５ )-cycloalkyl may be substituted with one or more fluoros, or R ｃ and R １ together with the atoms to which they are attached form a fused cyclopropyl ring, or R ｃ and R ｄ together are oxo (=O) or methylene (=CH ２ )), or R ｃ and R ｄ together with the atoms to which they are attached form spiro(C ３ -C ５ )-cycloalkyl, R １ is substituted with one or more fluoros and is (C １ -C ３ )alkyl, Each R ２ is independently selected from the group consisting of halo, cyano, hydroxy, ethynyl, (C １ -C ３ )alkylsulfonyl, cyclopropyl, (C １ -C ３ )alkyl, and (C １ -C ３ )alkoxy, and any cyclopropyl, (C １ -C ３ )alkyl, and (C １ -C ３ )alkoxy may be substituted with one or more fluoros, R ４ is (C １ -C ７ )alkyl, (C ３ -C ７ )cycloalkyl, phenyl, or 6-membered heteroaryl, and any R ４ may be substituted with one or more R ｘ s, Each R ｘ is independently selected from the group consisting of hydroxy, halo, cyano, (C １ -C ５ -C １ -C ５ -C ３ -C ５ -C ３ -C ７ -C １ -C ５ -C １ -C ５ -C ３ -C ５ -C ３ -C ５ -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C -C [[ A is 0, 1, or 2, said compound or a salt thereof.

2. L １ is CR ａ R ｂ and L ２ is CR ｃ R ｄ and R ａ is H, halo, cyano, (C １ -C ５ -alkyl, (C １ -C ５ -alkoxy, (C ３ -C ５ -cycloalkyloxy, or (C ３ -C ５ -cycloalkyl, and any (C １ -C ５ -alkyl, (C １ -C ５ -alkoxy, (C ３ -C ５ -cycloalkyloxy, and (C ３ -C ５ -cycloalkyl may be substituted with one or more fluoros, R ｂ is H, halo, cyano, (C １ -C ５ -alkyl, (C １ -C ５ -alkoxy, or (C ３ -C ５ -cycloalkyl, and any (C １ -C ５ -alkyl, (C １ -C ５ -alkoxy, and (C ３ -C ５ -cycloalkyl may be substituted with one or more fluoros, or R ａ and R ｂ together are oxo (=O) methylene (=CH ２ ), or R ａ and R ｂ together with the atoms to which they are attached form spiro(C ３ -C ５ -cycloalkyl, R ｃ is H, halo, cyano, (C １ -C ５ alkyl, (C １ -C ５ alkoxy, (C ３ -C ５ cycloalkyloxy, or (C ３ -C ５ cycloalkyl, and any (C １ -C ５ alkyl, (C １ -C ５ alkoxy, (C ３ -C ５ cycloalkyloxy, and (C ３ -C ５ cycloalkyl may be substituted with one or more fluoro, R ｄ is H, halo, cyano, (C １ -C ５ alkyl, (C １ -C ５ alkoxy, or (C ３ -C ５ cycloalkyl, and any (C １ -C ５ alkyl, (C １ -C ５ alkoxy, and (C ３ -C ５ cycloalkyl may be substituted with one or more fluoro, or R ｃ and R １ together with the atom to which they are attached form a fused cyclopropyl ring, or R ｃ and R ｄ together are oxo (=O) or methylene (=CH ２ ), or R ｃ and R ｄ together with the atom to which they are attached form spiro(C ３ -C ５ cycloalkyl, R ａ 、R ｂ 、R ｃ 、and R ｄ at least one of which is other than H, R １ is H, hydroxy, cyano, halo, methoxycarbonyl, cyclopropyl, (C １ -C ３ )alkyl, or (C １ -C ３ )alkoxy, and any cyclopropyl, (C １ -C ３ )alkyl, and (C １ -C ３ )alkoxy may be substituted with one or more groups independently selected from hydroxy, (C １ -C ３ )alkoxy, benzyloxy, cyano, and fluoro, or R １ and R ｃ together with the atom to which they are attached form a fused cyclopropyl ring, Each R ２ is independently selected from the group consisting of halo, cyano, hydroxy, ethynyl, (C １ -C ３ )alkylsulfonyl, cyclopropyl, (C １ -C ３ )alkyl, and (C １ -C ３ )alkoxy, and any cyclopropyl, (C １ -C ３ )alkyl, and (C １ -C ３ )alkoxy may be substituted with one or more fluoros, R ４ is (C １ -C ７ )alkyl, (C ３ -C ７ )cycloalkyl, phenyl, or 6-membered heteroaryl, and any R ４ may be substituted with one or more R ｘ s, Each R ｘ is independently selected from the group consisting of hydroxy, halo, cyano, (C １ -C ５ )alkyl, (C １ )alkoxy, and (C ５ )cycloalkyl, and any (C ３ )alkyl, (C ７ )alkoxy, and (C １ )cycloalkyl may be substituted with one or more groups independently selected from halo, cyano, (C ５ )alkyl, and hydroxy, and １ -C ５ )alkoxy, and (C ３ )cycloalkyl may be substituted with one or more groups independently selected from halo, cyano, (C ５ )alkyl, and hydroxy, and １ -C ５ )alkyl may be substituted with one or more groups independently selected from halo, cyano, (C A is 0, 1, or 2, the compound or salt according to Claim 1.

3. L １ is CR ａ R ｂ and L ２ is CR ｃ R ｄ and R ａ is H, halo, cyano, (C １ -C ５ alkyl, (C １ -C ５ alkoxy, (C ３ -C ５ cycloalkyloxy, or (C ３ -C ５ cycloalkyl, and any (C １ -C ５ alkyl, (C １ -C ５ alkoxy, (C ３ -C ５ cycloalkyloxy, and (C ３ -C ５ cycloalkyl may be substituted with one or more fluorines, R ｂ is H, halo, cyano, (C １ -C ５ alkyl, (C １ -C ５ alkoxy, or (C ３ -C ５ cycloalkyl, and any (C １ -C ５ alkyl, (C １ -C ５ alkoxy, and (C ３ -C ５ cycloalkyl may be substituted with one or more fluorines, or R ａ and R ｂ together form oxo (=O) methylene (=CH ２ ), or R ａ and R ｂ together with the atoms to which they are attached form spiro(C ３ -C ５ cycloalkyl, R ｃ is H, halo, cyano, (C １ -C ５ -C １ -C ５ -C ３ -C ５ -C ３ -C ５ -C １ -C ５ -C １ -C ５ -C ３ -C ５ -C ３ -C ５ -C ｄ is H, halo, cyano, (C １ -C ５ -C １ -C ５ -C ３ -C ５ -C １ -C ５ -C １ -C ５ -C ３ -C ５ -C ｃ and R １ together with the atom to which they are attached form a fused cyclopropyl ring, or R ｃ and R ｄ together are oxo (=O) or methylene (=CH ２ ), or R ｃ and R ｄ together with the atom to which they are attached form spiro(C ３ -C ５ ), R １ is substituted with one or more fluoro atoms in (C １ -C ３ )alkyl, Each R ２ is independently selected from the group consisting of halo, cyano, hydroxy, ethynyl, (C １ -C ３ )alkylsulfonyl, cyclopropyl, (C １ -C ３ )alkyl, and (C １ -C ３ )alkoxy, and any cyclopropyl, (C １ -C ３ )alkyl, and (C １ -C ３ )alkoxy may be substituted with one or more fluoros, R ４ is (C １ -C ７ )alkyl, (C ３ -C ７ )cycloalkyl, phenyl, or 6-membered heteroaryl, and any R ４ may be substituted with one or more R ｘ s, Each R ｘ is independently selected from the group consisting of hydroxy, halo, cyano, (C １ -C ５ )alkyl, (C １ -C ５ )alkoxy, and (C ３ -C ７ )cycloalkyl, and any (C １ -C ５ )alkyl, (C １ -C ５ )alkoxy, and (C ３ -C ５ )cycloalkyl may be substituted with one or more groups independently selected from halo, cyano, and hydroxy, and A is 0, 1, or 2, the compound or salt according to Claim 1.

4. L １ is CR ａ R ｂ and L ２ is CR ｃ R ｄ and R ａ is H, halo, cyano, (C １ -C ５ alkyl, (C １ -C ５ alkoxy, or (C ３ -C ５ cycloalkyl, and any (C １ -C ５ alkyl, (C １ -C ５ alkoxy, and (C ３ -C ５ cycloalkyl may be substituted with one or more fluorines, R ｂ is H, halo, cyano, (C １ -C ５ alkyl, (C １ -C ５ alkoxy, or (C ３ -C ５ cycloalkyl, and any (C １ -C ５ alkyl, (C １ -C ５ alkoxy, and (C ３ -C ５ cycloalkyl may be substituted with one or more fluorines, or R ａ and R ｂ together form oxo (=O) methylene (=CH ２ ), or R ａ and R ｂ together with the atoms to which they are attached form spiro (C ３ -C ５ cycloalkyl, R ｃ is H, halo, cyano, (C １ -C ５ )-alkyl, (C １ -C ５ )-alkoxy, or (C ３ -C ５ )-cycloalkyl, and any (C １ -C ５ )-alkyl, (C １ -C ５ )-alkoxy, and (C ３ -C ５ )-cycloalkyl may be substituted with one or more fluoros, R ｄ is H, halo, cyano, (C １ -C ５ )-alkyl, (C １ -C ５ )-alkoxy, or (C ３ -C ５ )-cycloalkyl, and any (C １ -C ５ )-alkyl, (C １ -C ５ )-alkoxy, and (C ３ -C ５ )-cycloalkyl may be substituted with one or more fluoros, or R ｃ and R １ together with the atoms to which they are attached form a fused cyclopropyl ring, or R ｃ and R ｄ together are oxo (=O) or methylene (=CH ２ ), or R ｃ and R ｄ together with the atoms to which they are attached form spiro(C ３ -C ５ )-cycloalkyl, R ａ 、R ｂ 、R ｃ 、and R ｄ at least one of which is other than H, R １ is H, hydroxy, cyano, halo, methoxycarbonyl, cyclopropyl, (C １ -C ３ )alkyl, or (C １ -C ３ )alkoxy, and any cyclopropyl, (C １ -C ３ )alkyl, and (C １ -C ３ )alkoxy may be substituted with one or more groups independently selected from hydroxy, (C １ -C ３ )alkoxy, benzyloxy, cyano, and fluoro, or R １ and R ｃ together with the atom to which they are attached form a fused cyclopropyl ring, Each R ２ is independently selected from the group consisting of halo, cyano, hydroxy, ethynyl, (C １ -C ３ )alkylsulfonyl, cyclopropyl, (C １ -C ３ )alkyl, and (C １ -C ３ )alkoxy, and any cyclopropyl, (C １ -C ３ )alkyl, and (C １ -C ３ )alkoxy may be substituted with one or more fluorines, R ４ is (C １ -C ７ )alkyl, (C ３ -C ７ )cycloalkyl, phenyl, or 6-membered heteroaryl, and any R ４ may be substituted with one or more R ｘ ; Each R ｘ is independently selected from the group consisting of hydroxy, halo, cyano, (C １ -C ５ )alkyl, (C １ -C ５ )alkoxy, and (C ３ -C ７ )cycloalkyl, and any (C １ -C ５ )alkyl, (C １ -C ５ )alkoxy, and (C ３ -C ５ )cycloalkyl may be substituted with one or more groups independently selected from halo, cyano, (C １ -C ５ )alkyl, and hydroxy, and A is 0, 1, or 2, the compound or salt according to Claim 1, or L １ is CR ａ R ｂ and L ２ is CR ｃ R ｄ and R ａ is H, halo, cyano, (C １ -C ５ )-alkyl, (C １ -C ５ )-alkoxy, or (C ３ -C ５ )-cycloalkyl, and any (C １ -C ５ )-alkyl, (C １ -C ５ )-alkoxy, and (C ３ -C ５ )-cycloalkyl may be substituted with one or more fluorines, R ｂ is H, halo, cyano, (C １ -C ５ )-alkyl, (C １ -C ５ )-alkoxy, or (C ３ -C ５ )-cycloalkyl, and any (C １ -C ５ )-alkyl, (C １ -C ５ )-alkoxy, and (C ３ -C ５ )-cycloalkyl may be substituted with one or more fluorines, or R ａ and R ｂ together form oxo(=O)methylene(=CH ２ ), or R ａ and R ｂ together with the atoms to which they are attached form spiro(C ３ -C ５ )cycloalkyl, R ｃ is H, halo, cyano, (C １ -C ５ )-alkyl, (C １ -C ５ )-alkoxy, or (C ３ -C ５ )-cycloalkyl, and any (C １ -C ５ )-alkyl, (C １ -C ５ )-alkoxy, and (C ３ -C ５ )-cycloalkyl may be substituted with one or more fluoro, R ｄ is H, halo, cyano, (C １ -C ５ )-alkyl, (C １ -C ５ )-alkoxy, or (C ３ -C ５ )-cycloalkyl, and any (C １ -C ５ )-alkyl, (C １ -C ５ )-alkoxy, and (C ３ -C ５ )-cycloalkyl may be substituted with one or more fluoro, or R ｃ and R １ together with the atom to which they are attached form a fused cyclopropyl ring, or R ｃ and R ｄ together are oxo (=O) or methylene (=CH ２ ), or R ｃ and R ｄ together with the atom to which they are attached form spiro(C ３ -C ５ )-cycloalkyl, R １ is substituted with one or more fluoro groups and is (C １ -C ３ )alkyl, Each R ２ is independently selected from the group consisting of halo, cyano, hydroxy, ethynyl, (C １ -C ３ ) alkylsulfonyl, cyclopropyl, (C １ -C ３ ) alkyl, and (C １ -C ３ ) alkoxy, and any cyclopropyl, (C １ -C ３ ) alkyl, and (C １ -C ３ ) alkoxy may be substituted with one or more fluoros. R ４ is (C １ -C ７ )alkyl, (C ３ -C ７ )cycloalkyl, phenyl, or 6-membered heteroaryl, and any R ４ may be substituted with one or more R ｘ s, Each R ｘ is independently selected from the group consisting of hydroxy, halo, cyano, (C １ -C ５ )alkyl, (C １ -C ５ )alkoxy, and (C ３ -C ７ )cycloalkyl, and any (C １ -C ５ )alkyl, (C １ -C ５ )alkoxy, and (C ３ -C ５ )cycloalkyl may be substituted with one or more groups independently selected from halo, cyano, and hydroxy, and A is 0, 1, or 2, the compound or salt according to Claim 1.

5. L １ is CR ａ R ｂ and L ２ is CR ｃ R ｄ and R ａ is H, halo, cyano, (C １ -C ５ ), alkyl, (C １ -C ５ ), alkoxy, or (C ３ -C ５ ), cycloalkyl, and any (C １ -C ５ ), alkyl, (C １ -C ５ ), alkoxy, and (C ３ -C ５ ), cycloalkyl may be substituted with one or more fluoros, R ｂ is H, halo, cyano, (C １ -C ５ ), alkyl, (C １ -C ５ ), alkoxy, or (C ３ -C ５ ), cycloalkyl, and any (C １ -C ５ ), alkyl, (C １ -C ５ ), alkoxy, and (C ３ -C ５ ), cycloalkyl may be substituted with one or more fluoros, or R ａ and R ｂ together are oxo (=O) methylene (=CH ２ ), or R ａ and R ｂ together with the atoms to which they are attached form spiro(C ３ -C ５ ), cycloalkyl spiro(C ３ -C ５ ), cycloalkyl, R ｃ is H, halo, cyano, (C １ -C ５ -alkyl, (C １ -C ５ -alkoxy, or (C ３ -C ５ -cycloalkyl, and any (C １ -C ５ -alkyl, (C １ -C ５ -alkoxy, and (C ３ -C ５ -cycloalkyl may be substituted with one or more fluoro, R ｄ is H, halo, cyano, Rc(C １ -C ５ -alkyl, (C １ -C ５ -alkoxy, or (C ３ -C ５ -cycloalkyl, and any (C １ -C ５ -alkyl, (C １ -C ５ -alkoxy, and (C ３ -C ５ -cycloalkyl may be substituted with one or more fluoro, or R ｃ and R １ together with the atom to which they are attached form a fused cyclopropyl ring, or R ｃ and R ｄ together are oxo (=O) or methylene (=CH ２ ) and R ｃ and R ｄ together with the atom to which they are attached form spiro(C ３ -C ５ )cycloalkyl spiro(C ３ -C ５ )cycloalkyl, R ａ 、 R ｂ 、 R ｃ 、 and R ｄ at least one of which is other than H, R １ is H, hydroxy, cyano, halo, methoxycarbonyl, cyclopropyl, (C １ -C ３ )alkyl, or (C １ -C ３ )alkoxy, and any cyclopropyl, (C １ -C ３ )alkyl, and (C １ -C ３ )alkoxy may be substituted with one or more groups independently selected from hydroxy, (C １ -C ３ )alkoxy, benzyloxy, cyano, and fluoro, or R １ and R ｃ together with the atom to which they are attached form a fused cyclopropyl ring, Each R ２ is independently selected from the group consisting of halo, cyano, hydroxy, ethynyl, (C １ -C ３ )alkylsulfonyl, cyclopropyl, (C １ -C ３ )alkyl, and (C １ -C ３ )alkoxy, and any cyclopropyl, (C １ -C ３ )alkyl, and (C １ -C ３ )alkoxy may be substituted with one or more fluoros. R ４ is (C １ -C ７ )alkyl, (C ３ -C ７ )cycloalkyl, phenyl, or 6-membered heteroaryl, and any R ４ may be substituted with one or more R ｘ s, Each R ｘ is independently selected from the group consisting of hydroxy, halo, cyano, (C １ -C ５ -alkyl, (C １ -C ５ -alkoxy, and (C ３ -C ７ -cycloalkyl, and any (C １ -C ５ -alkyl, (C １ -C ５ -alkoxy, and (C ３ -C ５ -cycloalkyl may be substituted with one or more groups independently selected from halo, cyano, (C １ -C ５ -alkyl, and hydroxy, and A is 0, 1, or 2, the compound or salt according to Claim 4.

6. L １ is CR ａ R ｂ and L ２ is CR ｃ R ｄ and R ａ is H, halo, cyano, (C １ -C ５ ), alkyl, (C １ -C ５ ), alkoxy, or (C ３ -C ５ ), cycloalkyl, and any (C １ -C ５ ), alkyl, (C １ -C ５ ), alkoxy, and (C ３ -C ５ ), cycloalkyl may be substituted with one or more fluoros, R ｂ is H, halo, cyano, (C １ -C ５ ), alkyl, (C １ -C ５ ), alkoxy, or (C ３ -C ５ ), cycloalkyl, and any (C １ -C ５ ), alkyl, (C １ -C ５ ), alkoxy, and (C ３ -C ５ ), cycloalkyl may be substituted with one or more fluoros, or R ａ and R ｂ together are oxo (=O) methylene (=CH ２ ), or R ａ and R ｂ together with the atoms to which they are attached form spiro(C ３ -C ５ ), cycloalkyl spiro(C ３ -C ５ ), cycloalkyl, R ｃ is H, halo, cyano, (C １ -C ５ ), alkyl, (C １ -C ５ ), alkoxy, or (C ３ -C ５ ), cycloalkyl, where any (C １ -C ５ ), alkyl, (C １ -C ５ ), alkoxy, and (C ３ -C ５ ) cycloalkyl may be substituted with one or more fluoro, R ｄ is H, halo, cyano, Rc(C １ -C ５ ), alkyl, (C １ -C ５ ), alkoxy, or (C ３ -C ５ ), cycloalkyl, where any (C １ -C ５ ), alkyl, (C １ -C ５ ), alkoxy, and (C ３ -C ５ ) cycloalkyl may be substituted with one or more fluoro, or R ｃ and R １ together with the atom to which they are attached form a fused cyclopropyl ring, or R ｃ and R ｄ together are oxo (=O) or methylene (=CH ２ ), or R ｃ and R ｄ together with the atom to which they are attached form spiro(C ３ -C ５ ) cycloalkyl spiro(C ３ -C ５ ) cycloalkyl, R １ is substituted with one or more fluoro groups and is (C １ -C ３ )alkyl, Each R ２ is independently selected from the group consisting of halo, cyano, hydroxy, ethynyl, (C １ -C ３ )alkylsulfonyl, cyclopropyl, (C １ -C ３ )alkyl, and (C １ -C ３ )alkoxy, and any cyclopropyl, (C １ -C ３ )alkyl, and (C １ -C ３ )alkoxy may be substituted with one or more fluoros, R ４ is (C １ -C ７ )alkyl, (C ３ -C ７ )cycloalkyl, phenyl, or 6-membered heteroaryl, and any R ４ may be substituted with one or more R ｘ ; Each R ｘ is hydroxy, halo, cyano, (C １ -C ５ ) alkyl, (C １ -C ５ ) alkoxy, and (C ３ -C ７ )cycloalkyl; １ -C ５ ) alkyl, (C １ -C ５ ) alkoxy, and (C ３ -C ５ ) the cycloalkyl is optionally substituted with one or more groups independently selected from halo, cyano, and hydroxy; and A is 0, 1, or 2, the compound or salt according to Claim 4.

7. Formula (II): The compound or a salt thereof, wherein R ４ is (C ３ -C ７ ), cycloalkyl, phenyl, or 6-membered heteroaryl, and B is 0, 1, or 2, the compound or salt according to claim 4.

8. Formula (III): The compound or a salt thereof, wherein R ４ is (C ３ -C ７ ), cycloalkyl, phenyl, or 6-membered heteroaryl, and B is 0, 1, or 2, the compound or salt according to claim 4.

9. R ａ The compound or salt according to any one of claims 1 to 8, wherein R is H.

10. R ａ But halo, cyano, (C １ -C ５ ) alkyl, (C １ -C ５ ) alkoxy, and (C ３ -C ５ ) cycloalkyl; any (C １ -C ５ ) alkyl, (C １ -C ５ ) alkoxy, and (C ３ -C ５ 9. The compound or salt according to any one of claims 1 to 8, wherein cycloalkyl is optionally substituted with one or more fluoro.

11. R ａ is H, halo, cyano, CF ３ , CF ２ H, CFH ２ , OCF ３ , OCF ２ H, OCH ２ CF ３ , OCH ２ CF ２ H, (C １ -C ５ ), alkyl, or (C ３ -C ５ ), cycloalkyl, and the compound or salt according to any one of claims 1 to 8.

12. R ａ is halo, cyano, CF ３ , CF ２ H, CFH ２ , OCF ３ , OCF ２ H, OCH ２ CF ３ , OCH ２ CF ２ H, (C １ -C ５ ), alkyl, or (C ３ -C ５ ), cycloalkyl, the compound or salt according to any one of claims 1 to 8.

13. R ｂ The compound or salt according to any one of claims 1 to 12, wherein R is H.

14. R ｂ is halo, cyano, (C １ -C ５ -alkyl, (C １ -C ５ -alkoxy, and (C ３ -C ５ -cycloalkyl, and any (C １ -C ５ -alkyl, (C １ -C ５ -alkoxy, and (C ３ -C ５ -cycloalkyl may be substituted with one or more fluorines, the compound or salt according to any one of claims 1 to 12.

15. R ｂ is H, halo, cyano, CF ３ , CF ２ H, CFH ２ , OCF ３ , OCF ２ H, OCH ２ CF ３ , OCH ２ CF ２ H, (C １ -C ５ ), alkyl, or (C ３ -C ５ ) cycloalkyl, the compound or salt according to any one of claims 1 to 12.

16. R ｂ is halo, cyano, CF ３ , CF ２ H, CFH ２ , OCF ３ , OCF ２ H, OCH ２ CF ３ , OCH ２ CF ２ H, (C １ -C ５ ), alkyl, or (C ３ -C ５ ), cycloalkyl, the compound or salt according to any one of claims 1 to 12.

17. R ｃ The compound or salt according to any one of claims 1 to 16, wherein R is H.

18. R ｃ is selected from the group consisting of halo, cyano, (C １ -C ５ )alkyl, (C １ -C ５ )alkoxy, and (C ３ -C ５ )cycloalkyl, and any (C １ -C ５ )alkyl, (C １ -C ５ )alkoxy, and (C ３ -C ５ )cycloalkyl may be substituted with one or more fluorines, the compound or salt according to any one of claims 1 to 16.

19. R ｃ is H, halo, cyano, CF ３ , CF ２ H, CFH ２ , OCF ３ , OCF ２ H, OCH ２ CF ３ , OCH ２ CF ２ H, (C １ -C ５ ), alkyl, or (C ３ -C ５ ), cycloalkyl, the compound or salt according to any one of claims 1 to 16.

20. R ｃ is halo, cyano, CF ３ , CF ２ H, CFH ２ , OCF ３ , OCF ２ H, OCH ２ CF ３ , OCH ２ CF ２ H, (C １ -C ５ ), alkyl, or (C ３ -C ５ ), cycloalkyl, a compound or salt according to any one of claims 1 to 16.

21. R １ and R ｃ form a fused cyclopropyl ring together with the atoms to which they are attached, a compound or salt according to any one of claims 1 to 16.

22. R ｄ The compound or salt according to any one of claims 1 to 21, wherein R is H.

23. R ｄ is selected from the group consisting of halo, cyano, (C １ -C ５ -alkyl, (C １ -C ５ -alkoxy, and (C ３ -C ５ -cycloalkyl, and any (C １ -C ５ -alkyl, (C １ -C ５ -alkoxy, and (C ３ -C ５ -cycloalkyl) may be substituted with one or more fluorines, The compound or salt according to any one of claims 1 to 21.

24. R ｄ is H, halo, cyano, CF ３ , CF ２ H, CFH ２ , OCF ３ , OCF ２ H, OCH ２ CF ３ , OCH ２ CF ２ H, (C １ -C ５ ), alkyl, or (C ３ -C ５ ), cycloalkyl, the compound or salt according to any one of claims 1 to 21.

25. R ｄ is halo, cyano, CF ３ CF ２ H, CFH ２ OCF ３ OCF ２ H, OCH ２ CF ３ OCH ２ CF ２ H, (C １ -C ５ alkyl, or (C ３ -C ５ cycloalkyl), a compound or salt according to any one of claims 1 to 21.

26. R ｃ and R ｄ together with the atoms to which they are attached form a spiro(C ３ -C ５ ) cycloalkyl, a compound or a salt according to any one of claims 1 to 16.

27. R １ is selected from the group consisting of cyano, halo, methoxycarbonyl, cyclopropyl, (C １ -C ３ )alkyl, and (C １ -C ３ )alkoxy, and any cyclopropyl, (C １ -C ３ )alkyl, and (C １ -C ３ )alkoxy may be substituted with one or more groups independently selected from hydroxy, (C １ -C ３ )alkoxy, benzyloxy, and fluoro, a compound or salt according to any one of claims 1 to 20 and 22 to 26.

28. R １ is H, methyl, cyclopropyl, CF ３ , CF ２ H, CFH ２ , OCF ３ , or OCF ２ H, and is the compound or salt according to any one of claims 1 to 20 and 22 to 26.

29. R １ The compound or salt according to any one of claims 1 to 20 and 22 to 26, wherein R is hydroxy, hydroxymethyl, or cyanomethyl.

30. R １ The compound or salt according to any one of claims 1 to 20 and 22 to 26, wherein R is methyl.

31. R １ The compound or salt according to any one of claims 1 to 20 and 22 to 26, wherein R is H.

32. said group: is selected from the group consisting of: the compound or salt according to any one of Claims 1 to 31.

33. A is 0, the compound or salt according to any one of Claims 1 to 31.

34. A is 1, the compound or salt according to any one of Claims 1 to 31.

35. R ２ The compound or salt according to claim 34, wherein R is cyano.

36. R ２ The compound or salt according to claim 34, wherein R is 6-cyano.

37. R ４ is phenyl which may be substituted with one or more R ｘ The compound or salt according to any one of claims 1 to 36, wherein R

38. R ４ is as follows: selected from the group consisting of: the compound according to any one of Claims 1 to 36.

39. R ４ is a pyrimidin-2-yl which may be substituted with one or more R ｘ The compound or salt according to any one of claims 1 to 36, wherein R

40. R ４ is as follows: selected from the group consisting of: the compound according to any one of Claims 1 to 36.

41. R ４ may be replaced by one R ｘ and may be (C １ -C ７ ) alkyl, the compound according to any one of claims 1 to 36.

42. R ４ may be replaced by one R ｘ and may be (C ３ -C ７ ) cycloalkyl, the compound according to any one of claims 1 to 36.

43. R ４ is a 6-membered heteroaryl which may be substituted with one R ｘ The compound according to any one of claims 1 to 36

44. R ｘ is C optionally substituted with one or more groups independently selected from halo, cyano, and hydroxy １ -C ５ alkyl, a compound according to any one of claims 41 to 43

45. R ｘ is C substituted with hydroxy １ -C ５ alkyl, the compound according to any one of claims 41 to 43.

46. R ｘ The compound according to any one of claims 41 to 43, wherein R is 2-hydroxy-2-methylethyl.

47. Formula IV or V: a compound or a salt thereof, wherein B is 0, 1, or 2, the compound or salt according to Claim 1.

48. Formula IV or Va: a compound or a salt thereof, wherein B is 0, 1, or 2, the compound or salt according to Claim 1.

49. Formula VI or VII: a compound or a salt thereof, the compound or salt according to Claim 1.

50. R ｃ is fluoro and R ｄ is H or fluoro, the compound or salt according to claim 1.

51. as follows: The compound or salt according to claim 1, selected from the group consisting of the same and salts thereof.

52. The following: The compound or salt according to claim 1, selected from the group consisting of the same and salts thereof.

53. The following: The compound or salt according to claim 1, selected from the group consisting of the same and salts thereof.

54. The following: The compound or salt according to claim 1, selected from the group consisting of the same and salts thereof.

55. The compound or salt according to claim 1, or a salt thereof.

56. The compound or salt according to claim 1, or a salt thereof.

57. A pharmaceutical composition comprising the compound or pharmaceutically acceptable salt according to any one of claims 1 to 56 and a pharmaceutically acceptable excipient.

58. A method for treating a condition related to TRPV4, comprising administering the compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 56.

59. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 56 for use in the treatment of a condition related to TRPV4.

60. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 56 for prophylactic or therapeutic treatment of a condition related to TRPV4.

61. Use of the compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 56 in the manufacture of a medicament for treating a condition related to TRPV4.

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