EA043804B1 - HISTONE DACETYLASE INHIBITORS - Google Patents

Description

Related applications

This application claims benefit from U.S. Provisional Patent Application No. 62/697,498, filed July 13, 2018, the entire contents of which are incorporated herein by reference.

State of the art

Histone deacetylase (HDAC) inhibitors have been shown to modulate transcription and induce cell growth arrest, differentiation, and apoptosis. HDAC inhibitors also enhance the cytotoxic effects of therapeutic agents used in the treatment of cancer, including radiation and chemotherapy drugs. Marks, P., Rifkind, R.A., Richon, V.M., Breslow, R., Miller, T., Kelly, W.K., Histone deacetylases and cancer: causes and therapies, Nat. Rev. Cancer, 1, 194-202, (2001); and Marks, P.A., Richon, V.M., Miller, T., Kelly, W.K., Histone deacetylase inhibitors, Adv. Cancer Res., 91, 137-168, (2004). Moreover, recent evidence suggests that transcriptional dysregulation may contribute to the molecular pathogenesis of several neurodegenerative diseases, such as Huntington's disease, spinal muscular atrophy, amyotrophic lateral sclerosis, and ischemia. Langley, B., Gensert, J.M., Beal, M.F., Ratan, R.R., Remodeling chromatin and stress resistance in the central nervous system: histone deacetylase inhibitors as novel and broadly effective neuroprotective agents, Curr. Drug Targets CNS Neurol. Disord., 4, 41-50, (2005). A recent review summarized evidence that aberrant histone acetyltransferase (HAT) and histone deacetylase (HDAC) activities may represent a common underlying mechanism contributing to neurodegeneration. Moreover, using a mouse model of depression, Nestler recently highlighted the therapeutic potential of histone deacetylation inhibitors (HDAC5) in depression. Tsankova, N.M., Berton, O., Renthal, W., Kumar, A., Neve, R.L., Nestler, E.J., Sustained hippocampal chromatin regulation in a mouse model of depression and antidepressant action, Nat. Neurosci., 9, 519-525 (2006).

There are 18 known human histone deacetylases, grouped into four classes based on the structure of their accessory domains. Class I includes HDAC1, HDAC2, HDAC3, and HDAC8 and shares homology with yeast RPD3. HDAC4, HDAC5, HDAC7 and HDAC9 belong to class IIa and have homology with yeast. HDAC6 and HDAC 10 contain two catalytic sites and belong to class IIb. Class III (sirtuins) includes SIRT1, SIRT2, SIRT3, SIRT4, SIRT5, SIRT6 and SIRT7. HDAC11 is another recently identified member of the HDAC family and has conserved residues in the catalytic center that are common to class I and class II deacetylases and are sometimes classified as class IV.

In contrast, HDACs have been shown to be potent negative regulators of long-term memory processes. Nonspecific HDAC inhibitors improve synaptic plasticity as well as long-term memory (Levenson et al., 2004, J. Biol. Chem., 279:40545-40559; Lattal et al., Behav. Neurosci., 121:1125-1131; Vecsey et al., Behav. Neurosci., 121:1125-1131 al., 2007, J. Neurosci., 27:6128; Bredy, 2008, Learn. Mem., 15:460-467; Guan et al., 2009, Nature, 459:55-60; Malvaez et al., 2010 , Biol Psychiatry 67:36–43; Roozendaal et al. 2010 J Neurosci 30:5037–5046). For example, HDAC inhibition can convert a learning event that does not result in long-term memory into a learning event that does result in significant long-term memory (Stefanko et al., 2009, Proc. Natl. Acad. Sci., USA, 106:9447- 9452). In addition, HDAC inhibition may also generate a form of long-term memory that persists beyond the point where normal memory fails. HDAC inhibitors have been shown to improve cognitive deficits in genetic models of Alzheimer's disease (Fischer et al., 2007, Nature, 447:178-182; Kilgore et al., 2010, Neuropsychopharmacology, 35:870-880). These data suggest that memory modulation through HDAC inhibition has significant therapeutic potential for many memory and cognitive disorders.

The role of individual HDACs in long-term memory has now been examined in two recent studies. Kilgore et al., 2010, Neuropsychopharmacology, 35:870-880, showed that nonspecific HDAC inhibitors, such as sodium butyrate, inhibit class I HDACs (HDAC1, HDAC2, HDAC3, HDAC8) with little effect on family members HDAC class IIa (HDAC4, HDAC5, HDAC7, HDAC9). This suggests that class I HDAC inhibition may be critical for the improvements in cognitive function observed in many studies. Indeed, forebrain- and neuron-specific overexpression of HDAC2, but not HDAC1, reduces dendritic spine density, synaptic density, synaptic plasticity, and memory formation (Guan et al., 2009, Nature, 459:55-60). In contrast, HDAC2 knockout mice exhibited increased synaptic density, increased synaptic plasticity, and increased dendrite density in neurons. These HDAC2-deficient mice also demonstrated enhanced learning and memory in an arsenal of learning behavioral paradigms. This study demonstrates that HDAC2 is a key regulator of synaptogenesis and synaptic plasticity. In addition, Guan et al. showed that chronic treatment of mice with SAHA (an HDAC inhibitor 1, 2, 3, 6, 8) reproduced the effects observed in HDAC2-deficient mice and reversed cognitive impairment in HDAC2-overexpressing mice.

HDAC2 inhibition (selective or in combination with other class I HDAC inhibition)

- 1 043804 is an attractive therapeutic target. Such inhibition has the potential to improve cognition and facilitate learning by increasing synaptic and dendritic density in neuronal cell populations. In addition, HDAC2 inhibition may also be therapeutically useful in treating a wide range of other diseases and disorders.

Disclosure of the invention

This application presents compounds of formula III or III

R ⁴ (III); or ^R 3 (Sha);

and pharmaceutically acceptable salts and compositions thereof, wherein X, R ¹ , R ² , R ³ , R ⁴ , and q are as described herein.

Also disclosed is the use of the above compound for inhibiting histone deacetylase (HDAC) activity in a subject, as well as for treating a condition in a subject selected from a neurological disorder, a cognitive disorder or impairment, a fungal infection, an inflammatory disease, a mental disorder, and a neoplastic disease.

Detailed Description of the Invention

1. General description of connections.

This application provides a compound of formula I

or a pharmaceutically acceptable salt thereof, wherein ring A is phenyl or thiophenyl;

X represents (CR ^a R ^b ) _t , O or NR ⁵ ;

q and t are each independently 0, 1, 2 or 3;

R ¹ represents phenyl or heteroaryl, each of which is optionally substituted with 1 to 3 groups selected from R ^c ;

R ² represents halogen, (C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )alkoxy or OH;

R ³ represents hydrogen or halogen;

R ⁴ is halogen if ring A is phenyl, and R ⁴ is hydrogen if ring A is thiophenyl;

R ⁵ represents hydrogen, (C ₁ -C ₄ )alkyl or (C ₁ -C ₄ )alkylO(C ₁ -C ₄ )alkyl;

R ^a and R ^b are each independently hydrogen, (C ₁ -C ₄ )alkyl, halogen(C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )alkoxy or halogen; And

R ^c represents halogen, (C ₁ -C ₄ )alkyl, halogen(C ₁ -C ₄ )alkyl, (C ₁ -C ₄ .)alkoxy, halogen(C ₁ C ₄ )alkoxy, (C ₁ -C ₄ )alkylO(C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )alkylNH(C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )alkylN((C ₁ -C ₄ )alkyl) ₂ , -( C ₁ -C ₄ )alkylheteroaryl or -(C ₁ -C ₄ )alkylheterocyclyl, wherein said heteroaryl and heterocyclyl are each optionally and independently substituted with 1 to 3 groups selected from (C ₁ -C ₄ )alkyl, halogen( C ₁ C ₄ )alkyl, (C ₁ -C ₄ )alkoxy and halogen.

The disclosed compounds and compositions modulate histone deacetylases (HDACs) (see, e.g., Tables 2 and 3) and are suitable for a variety of therapeutic applications, such as, for example, in the treatment of neurological disorders, memory or cognitive function disorders or impairments, learning disabilities (disorder of learning extinction), fungal diseases or infections, inflammatory diseases, hematological diseases, neoplastic diseases, mental disorders and memory loss. Some of the compounds described herein have increased inhibitory activity in cell lysate assays compared to direct reference compounds. For example, it was found that the introduction of aromatic substituents at the 3-position of pyrrolidine (e.g., phenyl and heteroaryl variables for R ¹ ) resulted in a significant increase in efficiency in the HDAC2 SH-SY5Y cell lysate assay compared to analogues having a non-aromatic substitution at the 3-position of pyrrolidine. See, for example, table. 4, where compounds 19 and 20, each of which has an aromatic pyrimidinyl in R ¹ , have greater

- 2 043804 efficiency than non-cyclic, non-aromatic compounds of comparison A-C.

2. Definitions.

When used in connection with a description of a chemical group that may have multiple attachment sites, a hyphen (-) indicates the location where that group is attached to the variable for which it is defined. For example, -(C ₁ -C ₄ )alkylheteroaryl and -(C ₁ -C ₄ )alkylheterocyclyl mean that the site of attachment is at the (C ₁ -C ₄ )alkyl residue.

The terms halo and halogen refer to an atom selected from fluorine (fluoro, -F), chlorine (chloro, -Cl), bromine (bromo, -Br) and iodine (iodo, -I).

The term alkyl, when used alone or as part of a larger moiety such as a haloalkyl, means a saturated straight-chain or branched monovalent hydrocarbon radical. Unless otherwise specified, an alkyl group usually contains 1-6 carbon atoms, i.e. (C ₁ -C ₆ )alkyl.

The term haloalkyl includes mono-, poly- and perhaloalkyl groups, where the halogens are independently selected from fluorine, chlorine, bromine and iodine.

The term alkoxy means an alkyl radical united through a linking oxygen atom represented by -O-alkyl. For example, (C ₁ -C ₄ )alkoxy includes methoxy, ethoxy, propoxy and butoxy.

The term haloalkoxy is a haloalkyl group attached to another moiety via an oxygen atom, for example, but not limited to -OCHF ₂ or -OCF ₃ .

The term heteroaryl refers to a 5-12 membered (eg 5 or 6 membered) aromatic radical containing 1-4 heteroatoms selected from N, O and S. A heteroaryl group may be mono- or bicyclic. Monocyclic heteroaryl includes, for example, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, etc. Bicyclic heteroaryls include groups in which a monocyclic heteroaryl ring is fused to one or more aryl or heteroaryl rings. Non-limiting examples include indolyl, imidazopyridinyl, benzoxazolyl, benzoxodiazolyl, indazolyl, benzimidazolyl, benzthiazolyl, quinolyl, quinazolinyl, quinoxalinyl, pyrrolopyridinyl, pyrrolopyrimidinyl, pyrazolopyridinyl, thienopyridinyl, thienopyrimidinyl, indolizinyl, purinyl , naphthyridinyl and pteridinyl. It should be understood that when indicated, optional substituents on the heteroaryl group may be present at any substitutable position and include, for example, the position at which the heteroaryl is attached.

The term heterocyclyl means a 4-12 membered (e.g. 4-6 membered) saturated or partially unsaturated heterocyclic ring containing from 1 to 4 heteroatoms independently selected from N, O and S. The heterocyclyl ring can be monocyclic, bicyclic (e.g. bridged, fused or spirobicyclic ring) or tricyclic. The heterocyclyl ring can be attached to its side group at any heteroatom or carbon atom, resulting in a stable structure. Examples of such saturated or partially unsaturated heterocyclic radicals include, but are not limited to, tetrahydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, pyrrolidinyl, pyridinonyl, pyrrolidonyl, piperidinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, morpholinyl, dihydrofuranyl, dihydropyranyl, dihydropyridinyl, those trahydropyridinyl, dihydropyrimidinyl, oxetanyl, azetidinyl and tetrahydropyrimidinyl . The term heterocyclyl also includes, for example, unsaturated heterocyclic radicals fused to another unsaturated heterocyclic radical or an aryl or heteroaryl ring, such as, for example, tetrahydronaphthyridine, indolinone, dihydropyrrolotriazole, imidazopyrimidine, quinolinone, dioxaspiro-decane. It should also be understood that, when present, optional substituents on a heterocyclyl group may be present at any substitutable position and include, for example, the position at which the heterocyclyl is attached (eg, in the case of an optionally substituted heterocyclyl or a heterocyclyl that is optionally substituted).

The term fused refers to two rings that have two adjacent ring atoms to each other.

The term spiro refers to two rings that share a ring atom (such as carbon).

The term bridging refers to two rings that share three ring atoms with each other.

Enantiomers are one type of stereoisomer that can arise from a chiral center or chiral centers. Enantiomers are pairs of stereoisomers whose mirror images cannot superpose, most often because they contain an asymmetrically substituted carbon atom or a carbon atom that acts as a chiral center(s). R and S represent the absolute configuration of substituents around one or more chiral carbon atoms, where each chiral center is assigned a prefix R or S depending on whether the configuration of the chiral center is right-handed (clockwise rotation) or left-handed (counterclockwise rotation) . If the turn is directed along

- 3 043804 clockwise or to the right relative to the chiral carbon, the prefix R indicates the direction to the right. If the rotation is counterclockwise or to the left with respect to the chiral carbon, the prefix S indicates the direction to the left.

If one enantiomer is named or depicted by structure, the enantiomer depicted or named is at least 60, 70, 80, 90, 99, or 99.9% optically pure by weight. The percentage of optical purity by mass is the ratio of the mass of the enantiomer to the mass of the enantiomer plus the mass of its optical isomer.

When a compound is depicted structurally without specifying the stereochemistry at the chiral center, the structure includes either the configuration at the chiral center or, alternatively, any mixture of configurations at stereoisomers of the chiral center.

The term racemate or racemic mixture means a combination of equimolar amounts of two enantiomers, where such mixtures do not exhibit optical activity, i.e. they do not rotate the plane of polarized light.

As used herein, the terms subject and patient may be used interchangeably and mean the mammal in need of treatment, e.g., domestic animals (e.g., dogs, cats, etc.), farm animals (e.g., cows, pigs, horses, sheep , goats, etc.) and laboratory animals (for example, rats, mice, guinea pigs, etc.). Typically the subject is a person in need of treatment.

Included in this application are pharmaceutically acceptable salts as well as neutral forms of the compounds described herein. For use in medicinal preparations, salts of the compounds are non-toxic pharmaceutically acceptable salts. Pharmaceutically acceptable salt forms include pharmaceutically acceptable acid/anionic or basic/cationic salts. Pharmaceutically acceptable basic/cationic salts include sodium, potassium, calcium, magnesium, diethanolamine, n-methyl-O-glucamine, L-lysine, L-arginine, ammonium, ethanolamine, piperazine and triethanolamine salts. Pharmaceutically acceptable acid/anionic salts include, for example, acetate, benzenesulfonate, benzoate, bicarbonate, bitartrate, carbonate, citrate, dihydrochloride, gluconate, glutamate, glycolylarsanilate, hexylresorcinate, hydrobromide, hydrochloride, malate, maleylate, malonate, nitrate, stearate, succinate, sulfate, tartrate and tosylate.

The term pharmaceutically acceptable carrier refers to a non-toxic carrier, adjuvant or vehicle that does not interfere with the pharmacological activity of the compound with which it is formulated. Pharmaceutically acceptable carriers, adjuvants or bases that may be used in the compositions described herein include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffers such as phosphates, glycine, sorbic acid, potassium sorbate, mixtures of partial glycerides of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium acid phosphate, potassium acid phosphate, sodium chloride, zinc salts, colloidal silicon dioxide, magnesium trisilicate, polyvinylpyrrolidone, substances based on celluloses, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene polyoxypropylene block polymers, polyethylene glycol and lanolin.

The terms treat and treat refer to reversing, alleviating, reducing the likelihood of developing, or suppressing the progression of a disease or disorder, or one or more symptoms thereof, as described herein. In some embodiments, treatment may be administered after one or more symptoms have developed, i.e. therapeutic treatment. In other embodiments, treatment may be administered in the absence of symptoms. For example, treatment may be prescribed to a susceptible individual prior to the onset of symptoms (eg, in light of a history of symptoms and/or in light of genetic or other susceptibility factors), i.e. preventive treatment. Treatment can also be continued after symptoms have disappeared, for example to prevent or delay their recurrence. The term effective amount or therapeutically effective amount includes the amount of a compound described herein that will produce a biological or medical response in a subject, e.g., 0.01 to 100 mg/kg body weight/day of compound provided, e.g., 0.1 to 100 mg/kg body weight/day.

3. Description of illustrative connections.

In a first embodiment of the invention there is provided a compound of formula I

- 4 043804 or a pharmaceutically acceptable salt thereof, wherein the variables are as described above for Formula I.

In a second embodiment of the invention there is provided a compound of formula II

or a pharmaceutically acceptable salt thereof, wherein the variables are as described above for Formula I.

In a third embodiment of the invention, a compound of formula III or III is provided

or a pharmaceutically acceptable salt thereof, wherein the variables are as described above for Formula I.

In a fourth embodiment, q in any of formulas I, II, III, or IIIa is 0 or 1; a R ² is halogen if q is 1, where the other variables are as described above for Formula I.

In a fifth embodiment of the invention, the compound of formula I is a compound of formula IV or IVa:

or a pharmaceutically acceptable salt thereof, wherein the variables are as described above for Formula I.

In a sixth embodiment, R ³ in any one of Formulas I, II, III, IIIa, IV or IVa is halogen, the remaining variables being as described above for Formula I or the fourth embodiment. Alternatively, R ³ in any of Formulas I, II, III, IIIa, IV or IVa is fluorine, the remaining variables being as described above for Formula I or the fourth embodiment. As yet another alternative, R ³ in any of Formulas I, II, III, IIIa, IV or IVa is hydrogen, the remaining variables being as described above for Formula I or the fourth embodiment.

In the seventh embodiment, R ⁴ in any of formulas I, II, III, IIIa, IV or IVa is fluorine, the remaining variables being as described above for formula I or the fourth or sixth embodiment.

In the eighth embodiment, X in any of formulas I, II, III, IIIa, IV or IVa is (CR ^a R ^b )t, the remaining variables being as described above for formula I or for the fourth, sixth, or seventh embodiment of the invention.

In the ninth embodiment, both R ^a and R ^b in any of formulas I, II, III, IIIa, IV or IVa are hydrogen, the remaining variables being as described above for formula I or for the fourth, sixth, seventh , or the eighth embodiment of the invention.

In a tenth embodiment of the invention there is provided a compound of formula V or Va

- 5 043804

or a pharmaceutically acceptable salt thereof, wherein the variables are as described above for Formula I, or for the fourth, sixth, or seventh embodiment of the invention.

An eleventh embodiment of the invention provides a compound of formula VI or VIa

R ¹

R ¹

R ⁴ (VI); or ^R 3 (Via):

or a pharmaceutically acceptable salt thereof, wherein the variables are as described above for Formula I or for the fourth, sixth, seventh, or eighth embodiment of the invention.

A twelfth embodiment of the invention provides a compound of formula VII or VIIa

X ¹ R ¹

R ⁴ (VII); or ^R 3 (Vila);

or a pharmaceutically acceptable salt thereof, the remaining variables being as described above for Formula I or for the fourth, sixth, seventh, eighth, or ninth embodiment of the invention.

In a thirteenth embodiment, R ¹ in any of formulas I, II, III, IIIa, IV, IVa, V, Va, VI, VIa, VII or VIIa is phenyl or a 5-6 membered monocyclic heteroaryl, each of which is optional substituted with one or more groups selected from R ^c , the remaining variables being the same as described above for Formula I or the fourth, sixth, seventh, eighth, or ninth embodiment of the invention. Alternatively, R ¹ in any of formulas I, II, III, IIIa, IV, IVa, V, Va, VI, VIa, VII or VIIa is phenyl, pyridinyl, pyrazinyl, pyridazinyl or pyrimidinyl, each of which is optionally substituted with 1 -2 groups selected from R ^c , the remaining variables being as described above for Formula I or the fourth, sixth, seventh, eighth, or ninth embodiments of the invention. As a further alternative, R ¹ in any of formulas I, II, III, IIIa, IV, IVa, V, Va, VI, VIa, VII or VIIa is thiazolyl, thiadiazolyl, imidazolyl, pyrazolyl or oxazolyl, each of which is optional substituted with 1-2 groups selected from R ^c , the remaining variables being the same as described above for Formula I or the fourth, sixth, seventh, eighth, or ninth embodiments of the invention.

In a fourteenth embodiment, R ^c in any of formulas I, II, III, IIIa, IV, IVa, V, Va, VI, VIa, VII or VIIa is halogen, (C1-C4)alkyl or (C1-C4) alkylO(C1-C4)alkyl, the remaining variables being as described above for Formula I or for the fourth, sixth, seventh, eighth, ninth, or thirteenth embodiments of the invention. Alternatively, R ^c in any of formulas I, II, III, IIIa, IV, IVa, V, Va, VI, VIa, VII or VIIa is fluorine, methyl or CH2OCH ₃ , other variables being as described above for Formula I or for the fourth, sixth, seventh, eighth, ninth, or thirteenth embodiments of the invention. As a further alternative, R ^c in any of formulas I, II, III, IIIa, IV, IVa, V, Va, VI, VIa, VII or VIIa is halogen, halogen(C ₁ -C ₄ )αalkyl or (C ₁ -C ₄ )alkyl, the remaining variables being as described above for Formula I or for the fourth, sixth, seventh, eighth, ninth, or thirteenth embodiments of the invention. As another alternative, R ^c in any of

- 6 043804 formulas I, II, III, IIIa, IV, IVa, V, Va, VI, Via, VII or Vila is fluorine, methyl or CHF2, the remaining variables being as described above for formula I or fourth, sixth, seventh, eighth, ninth, or thirteenth embodiments of the invention.

In a fifteenth embodiment, the invention provides the compound described below in the Examples section. Included herein are pharmaceutically acceptable salts and free forms of the exemplified compounds.

4. Application, compositions and administration.

In some embodiments, the compounds and compositions described herein are useful for treating conditions associated with HDAC activity. Such conditions include, for example, the conditions described below.

Recent reports detail the importance of histone acetylation in central nervous system (CNS) functions such as neuronal differentiation, memory formation, drug addiction, and depression. (Citrome, Psychopharmacol. Bull., 2003, 37, Suppl. 2, 74-88; Johannessen, CNS Drug Rev., 2003, 9, 199-216; Tsankova et al., 2006, Nat. Neurosci., 9, 519 - 525). Thus, in one aspect, the present compounds and compositions may be useful in the treatment of a neurological disorder. Examples of neurological disorders include (i) chronic neurodegenerative diseases such as familial and sporadic amyotrophic lateral sclerosis (FALS and ALS, respectively), familial and sporadic Parkinson's disease, Huntington's disease, familial and sporadic Alzheimer's disease, multiple sclerosis, muscular dystrophy, olivopontocerebellar atrophy, multiple system atrophy, Wilson's disease, progressive supranuclear palsy, diffuse Lewy body disease, frontotemporal lobar degeneration (FTLD), corticodentatonigal degeneration, progressive familial myoclonic epilepsy, strionigral degeneration, torsion dystonia, essential tremor, Down syndrome, Gilles de la syndrome Tourette's disease, Hallervorden-Spatz disease, diabetic peripheral neuropathy, dementia pugilistica, AIDS-related dementia, age-related dementia, age-related memory loss, and neurodegenerative diseases associated with amyloidosis, such as those caused by prion protein (PrP), which is associated with transmissible spongiform encephalopathy ( Creutzfeldt-Jakob disease, Gerstmann-Straussler-Scheinker syndrome, scrapie and kuru), and diseases caused by excessive accumulation of cystatin C (hereditary cystatin C angiopathy); and (ii) acute neurodegenerative disorders such as traumatic brain injury (eg, surgery-related brain injury), cerebral edema, peripheral nerve injury, spinal cord injury, Leigh's disease, Guillain-Barré syndrome, lysosomal storage diseases such as lipofuscinosis, Alper's disease, restless legs syndrome, dizziness as a result of central nervous system degeneration; pathologies resulting from chronic alcohol or drug abuse, including, for example, degeneration of locus coeruleus and cerebellar neurons, drug-induced movement disorders; pathologies that occur with age, including degeneration of cerebellar and cortical neurons, leading to cognitive and motor impairment; and pathologies associated with chronic amphetamine abuse, including degeneration of basal ganglia neurons leading to movement disorders; pathological changes resulting from focal trauma such as stroke, focal ischemia, vascular insufficiency, hypoxic-ischemic encephalopathy, hyperglycemia, hypoglycemia, or direct trauma; pathologies that occur as a negative side effect of therapeutic drugs and treatments (for example, degeneration of neurons in the cingulate and entorhinal cortex in response to anticonvulsant doses of NMDA glutamate receptor antagonists) and dementia associated with Wernicke-Korsakoff.

Neurological disorders affecting sensory neurons include Friedreich's ataxia, diabetes, peripheral neuropathy, and retinal neuronal degeneration. Other neurological disorders include nerve damage or trauma associated with spinal cord damage. Neurological disorders of the limbic and cortical systems include cerebral amyloidosis, Pick's atrophy, and Rett syndrome. In another aspect, neurological disorders include mood disorders such as mood disorders and anxiety; social behavior disorders such as character defects and personality disorders; learning, memory and intelligence disorders such as mental retardation and dementia. Thus, in one aspect, the disclosed compounds and compositions may be useful in the treatment of schizophrenia, delirium, attention deficit disorder (ADD), schizoaffective disorder, Alzheimer's disease, Rubinstein-Taybi syndrome, depression, mania, attention deficit disorder, substance abuse disorder, dementia, agitation, apathy, anxiety, psychosis, personality disorders, bipolar disorders, unipolar affective disorder, obsessive-compulsive disorders, eating disorders, post-traumatic stress disorders, irritability, conduct disorders and disinhibition in adolescents.

Transcription is considered a key step in long-term memory processes (Alberini, 2009, Physiol. Rev., 89, 121-145). Transcription is promoted by specific chromatin modifications, such as histone acetylation, which modulate the interaction of histones with DNA (Kouzarides, 2007,

- 7 043804

Cell 128:693–705). Modifying enzymes such as histone acetyltransferases (HATs) and histone deacetylases (HDACs) regulate the acetylation state of histone tails. In general, histone acetylation promotes gene expression, whereas histone deacetylation leads to gene repression. Numerous studies have shown that active HAT, the cAMP response element activating protein CREB-binding protein (CBP), is required for long-term forms of synaptic plasticity and long-term memory (for review, see Barrett, 2008, Learn Mem, 15:460-467) . Thus, in one aspect, the present compounds and compositions may be useful for stimulating cognitive function and enhancing learning and memory formation.

The compounds and compositions described herein may also be useful for the treatment of fungal diseases or infections.

In another aspect, the compounds and compositions described herein may be useful for the treatment of inflammatory diseases such as stroke, rheumatoid arthritis, lupus erythematosus, ulcerative colitis and traumatic brain injury (Leoni et al., PNAS, 99(5), 2995-3000 (2002); Suuronen et al., J. Neurochem., 87, 407-416 (2003), Drug Discovery Today, 10:197-204 (2005)).

In yet another aspect, the compounds and compositions described herein can be used to treat cancer caused by proliferation of neoplastic cells. Such cancers include, for example, solid tumors, neoplasms, carcinomas, sarcomas, leukemias, lymphomas, and the like. In one aspect, cancers that can be treated with the compounds and compositions described herein include, but are not limited to: heart cancer, lung cancer, gastrointestinal cancer, genitourinary tract cancer, liver cancer, nervous system cancer, gynecological cancer, hematological cancer, skin cancer and adrenal cancer. In one aspect, the compounds and compositions described herein are useful for the treatment of cardiac cancer selected from sarcoma (angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyoma, fibroma, lipoma and teratoma. In another aspect, the compounds and compositions described herein are useful for the treatment of lung cancer selected from bronchogenic carcinoma (squamous cell, undifferentiated small cell, undifferentiated large cell adenocarcinoma), alveolar (bronchiolar) carcinoma, bronchial adenoma, sarcoma, lymphoma, chondromatous hamartoma and mesothelioma . In one aspect, the compounds and compositions described herein are useful for the treatment of cancer of the gastrointestinal tract selected from cancer of the esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach (carcinoma, lymphoma, leiomyosarcoma), pancreas (ductal adenocarcinoma). , glucoma, insulinoma, gastrinoma, carcinoid tumors, VIPoma), small intestine (adenocarcinoma, lymphoma, carcinoid tumors, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma) and colon (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma, leiomyoma) ohm ). In one aspect, the compounds and compositions described herein are useful for the treatment of cancers of the genitourinary tract selected from cancers of the kidney (adenocarcinoma, Wilm's tumor [nephroblastoma], lymphoma, leukemia), bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate (adenocarcinoma, sarcoma) and testicles (seminoma, teratoma, embryonal carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, interstitial cell carcinoma, fibroma, fibroadenoma, adenomatoid tumors, lipoma). In one aspect, the compounds and compositions described herein are useful for the treatment of liver cancer selected from hepatoma (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma and hemangioma.

In some embodiments, the compounds described herein relate to the treatment of bone cancer selected from osteosarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticulocellular sarcoma), multiple giant myeloma, malignant giant cell tumor chordoma, osteochondroma (osteocartilaginous exostoses), benign chondroma, chondroblastoma, chondromyxofibroma, osteoid osteoma and giant cell tumors. In one aspect, the compounds and compositions described herein are useful for the treatment of cancers of the nervous system selected from cancers of the skull (osteoma, hemangioma, granuloma, xanthoma, osteitis deformans), meninges (meningioma, meningiosarcoma, gliomatosis), brain (astrocytoma , medulloblastoma, glioma, ependymoma, germinoma [pinealoma], glioblastoma multiforme, oligodendroglioma, schwannoma, retinoblastoma, congenital tumors) and spinal cord (neurofibroma, meningioma, glioma, sarcoma).

In one aspect, the compounds and compositions described herein can be used for the treatment of gynecological cancer selected from cancer of the uterus (endometrial carcinoma), cervix (cervical carcinoma, precancerous cervical dysplasia), ovary (ovarian carcinoma [serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma], granulosathecal cell tumors, Sertoli-Leydig cell tumors, dysgerminoma, malignant teratoma), vulva (squamous cell carcinoma, intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, melanoma), boot cell carcinoma, squamous cell carcinoma, squamous cell carcinoma carcinoma, botryoid sarcoma ( embryonal rhabdomyosarcoma) and fallopian tubes (carcinoma).

- 8 043804

In one aspect, the compounds and compositions described herein are useful for the treatment of skin cancer selected from malignant melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, dysplastic nevi, lipomas, angiomas, dermatofibromas, keloids and psoriasis.

In one aspect, the compounds and compositions described herein can be used to treat adrenal cancer selected from neuroblastoma.

In one aspect, the compounds and compositions described herein can be used to treat cancer, which includes, but is not limited to, leukemias, including acute leukemias and chronic leukemias such as acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia leukemia (CLL), chronic myelogenous leukemia (CML) and hairy cell leukemia; lymphomas such as cutaneous T-cell lymphomas (CTCL), non-cutaneous peripheral T-cell lymphomas, human T-cell lymphotrophic virus (HTLV)-associated lymphomas such as adult T-cell leukemia/lymphoma (ATLL), Hodgkin's disease and non-Hodgkin's lymphomas , large cell lymphomas, diffuse large B-cell lymphomas (DLBCL); Burkitt's lymphoma; mesothelioma, primary lymphoma of the central nervous system (CNS); multiple myeloma; pediatric solid tumors such as brain tumors, neuroblastoma, retinoblastoma, elm B tumor, bone tumors and soft tissue sarcomas, advanced adult solid tumors such as head and neck cancer (eg cancer of the mouth, larynx and esophagus), genitourinary cancer (eg, prostate, bladder, kidney, uterine, ovarian, testicular, rectal, and colon cancer), lung cancer, breast cancer, pancreatic cancer, melanoma and other skin cancers, stomach cancer, brain tumors , liver cancer and thyroid cancer.

In one aspect, this application provides a method of treating a subject suffering from a neurological disorder, a disorder or impairment of memory or cognitive function, a learning disorder, a fungal disease or infection, an inflammatory disease, a hematological disease, a mental disorder, and a neoplastic disease, comprising administering to the subject an effective amount of a compound , described in this application, or a pharmaceutically acceptable salt thereof, or a composition containing a compound described in this application.

Also provided herein is a method of treating a subject suffering from (a) a disorder or impairment of cognitive function associated with Alzheimer's disease, posterior cortical atrophy, normal pressure hydrocephalus, Huntington's disease, seizure-induced memory loss, schizophrenia, Rubinstein-Taybi syndrome, Rett syndrome, depression, fragile X syndrome, dementia with Lewy bodies, vascular dementia, vascular cognitive impairment (VCI), Binswanger disease, frontotemporal lobar degeneration (FTLD), ADHD, dyslexia, major depressive disorder, bipolar disorder, and social, cognitive and learning disorders associated with autism, traumatic brain injury (TBI), chronic traumatic encephalopathy (CTE), multiple sclerosis (MS), attention deficit disorder, anxiety disorder, conditioned fear, panic disorder, obsessive-compulsive disorder, post-traumatic stress disorder (PTSD), phobia, social anxiety disorder, drug addiction recovery, age-related memory impairment (AAMI), age-related cognitive decline (ARCD), ataxia, Parkinson's disease or Parkinson's disease dementia; or (b) a hematological disease selected from acute myeloid leukemia, acute promyelocytic leukemia, acute lymphoblastic leukemia, chronic myelogenous leukemia, myelodysplastic syndromes and sickle cell anemia; or (c) a neoplastic disease; or (d) disorder of learning extinction, selected from fear extinction and post-traumatic stress disorder; or (e) hearing loss or hearing impairment; or (e) fibrotic diseases such as pulmonary fibrosis, renal fibrosis, cardiac fibrosis and scleroderma; or (g) bone pain in patients with cancer; or (h) neuropathic pain, comprising administering to the subject an effective amount of a compound described herein, or a pharmaceutically acceptable salt thereof, or a composition containing a compound described herein.

Also provided is a method of treating a subject suffering from Alzheimer's disease, Huntington's disease, frontotemporal dementia, Friedreich's ataxia, post-traumatic stress disorder (PTSD), Parkinson's disease, or drug addiction recovery, comprising administering to the subject an effective amount of a compound described herein, or a pharmaceutically acceptable salt, or composition containing a compound described in this application.

- 9 043804

Also provided is a compound described herein, or a pharmaceutically acceptable salt thereof, or a composition for the treatment of one or more of the described conditions.

Also provided is a compound described herein, or a pharmaceutically acceptable salt thereof, or a composition for the preparation of a medicament for the treatment of one or more of the described conditions.

Subjects suffering from one or more of the described conditions may also be selected prior to treatment with a compound described herein, or a pharmaceutically acceptable salt thereof, or the present composition.

The present disclosure also provides pharmaceutically acceptable compositions containing a compound described herein or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier. These compositions can be used to treat one or more of the conditions described above.

The compositions described herein can be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally, or through an implanted reservoir. The term parenteral as used herein includes subcutaneous, intravenous, intramuscular, intraarticular, intrasynovial, intrathoracic, intrathecal, intrahepatic, intralesional and intracranial injection or infusion methods. Included herein are liquid dosage forms, injectable dosage forms, solid dispersion forms, and dosage forms for topical or transdermal administration of the compound.

It should also be understood that the specific dosage and treatment regimen for any particular patient will depend on a variety of factors, including age, body weight, general health, gender, diet, timing of administration, elimination rate, drug combination, physician judgment, and the severity of the particular disease to be treated. The amount of compound present in the composition will also depend on the particular compound in the composition.

Examples

As shown in the Examples below, in some illustrative embodiments of the invention, compounds are prepared in accordance with the following general procedures. It should be understood that while the general methods describe the synthesis of certain compounds of the present invention, the following general methods and other methods known to one skilled in the art may be applied to all compounds and subclasses and species of each of these compounds as described herein .

General information.

The spots were visualized using UV light (254 and 365 nm). Purification by column and flash chromatography was performed using silica gel (200-300 mesh). Solvent systems are reported as solvent ratios.

NMR spectra were recorded on a Bruker 400 spectrometer (400 MHz). 1H chemical shifts are represented by δ (delta) values in ppm with tetramethylsilane (TMS,=0.00 ppm) as internal standard. See, for example, the data presented in table. 1.

LC-MS spectra were acquired on an Agilent 1200 series 6110 or 6120 mass spectrometer with ESI (+) ionization mode. See, for example, the data presented in table. 1.

Synthesis 1949-A. A mixture of 6-chloro-3-nitropyridin-2-amine (4.58 g, 26.4 mmol), 2,4-difluorophenylboronic acid (5.00 g, 31.7 mmol) and Cs ₂ CO ₃ (25.73 g, 79.2 mmol) in dioxane/H ₂ O (100 ml/10 ml) was treated with Pd(PPh ₃ ) ₄ (1.10 g, 0.95 mmol) under N2. The mixture was stirred at 100°C for 2 hours and then concentrated in vacuo. The residue was dissolved in EtOAc (200 ml) and half

- 10 043804 the prepared solution was washed with brine (100 ml x 3). The organic layer was dried over anhydrous Na2SO4 and then concentrated in vacuo. The residue was purified by silica gel column chromatography (PE:EAAc=7:1~5:1) to give 1949-A (4.0 g, 61%) as a yellow solid. MS 252.1 [M+H]+.

Synthesis 1949-B. A solution of 1949-A (4.0 g, 15.94 mmol) in pyridine (60 ml) was cooled to 0°C and then phenylcarbonochloridate (7.50 g, 47.81 mmol) was added dropwise. After addition was complete, the mixture was heated to 50°C and stirred at 50°C for 4 hours. The mixture was then concentrated in vacuo and the residue was purified by silica gel column chromatography (PE:DCM=3:2~1:1) to obtain 1949-B (7.1 g, 91%) as a yellow solid. MS 492.1 [M+H] ⁺ .

Synthesis of 1949-C Mixture of 1949-B (140 mg, 0.29 mmol), 2-pyrrolidin-3-yl-pyridine (51 mg, 0.34 mmol) and Cs ₂ CO ₃ (279 mg, 0.86 mol) in acetonitrile (5 ml) was stirred at room temperature for

h. The mixture was then diluted with water (10 ml) and extracted with EtOAc (10 mlx3). The combined organic layers were washed with brine (10 mlx3), dried over _{anhydrous Na2SO4} and then concentrated in vacuo. The residue was purified by Prep-TLC (DCM:EtOAc=1:1) to give 1949-C (70 mg, 57%) as a yellow solid. MS 426.2 [M+H] ⁺ .

Synthesis of Compound 1 A mixture of 1949-C (70 mg, 0.16 mmol) and Pd/C (70 mg) in MeOH/EtOAc (3 mL/3 mL) was stirred at room temperature for 1 h under H2 atmosphere. Pd/C was then removed by filtration through Celite, the filtrate was concentrated and the residue was purified by Prep-TLC (DCM:MeOH=20:1) to give Compound 1 (30 mg, 47%) as a yellow solid. MS 396.2 [M+H] ⁺ .

Compounds 2-6 were synthesized similarly using an appropriately substituted amine version of the reagent used for the synthesis of compound 1.

Compound 2. 40 mg, 54%, white solid.

Compound 3. 48 mg, 52%, white solid.

Compound 4. 70 mg, 69%, white solid.

Compound 5: 109 mg, 97%, white solid (derived from commercially available chiral building blocks).

Compound 6. 95 mg, 73%, gray solid (derived from commercially available chiral building blocks).

Example 2.

Synthesis 1981-A. A mixture of zinc dust (840 mg, 12.9 mmol) and celite (180 mg) in a sealed flask was heated under vacuum using a heat gun for 5 min. The flask was purged with N ₂ and cooled to room temperature. Anhydrous DMA (5.5 ml) was added to the mixture, followed by a mixture of TMSC1 and 1,2-dibromoethane (0.3 ml, v/v=7/5). The mixture was stirred at room temperature for 15 minutes under N ₂ atmosphere, after which a solution of tert-butyl-3-iodopyrrolidine-1-carboxylate (3.10 g, 10.4 mmol) in DMA (5.5 ml) was added. The resulting mixture was stirred at room temperature for 4 hours under N2 atmosphere, and then the mixture was used in the next step directly as 1981-A. The concentration of 1981-A was approximately 0.85 mol/L in DMA.

Synthesis 1981-B. A mixture of 2-bromopyrimidine (1.1 g, 6.92 mmol), CuI (197 mg, 1.04 mmol) and Pd(dppf) ₂ Cl ₂ (452 mg, 0.55 mmol) in DMA (10 ml) in atmosphere N ₂ was treated with 1981-A (10.0 ml). The resulting mixture was stirred at 85°C for 48 hours under N2, and then the mixture was diluted with water (50 ml) and extracted with EtOAc (20 ml x 3). The combined organic layers were washed with solution (20 mlx3), dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified by silica gel column chromatography. (PE:EtOAc=10:1~1:1) to give 1981-B (320 mg, 19%) as a yellow oil. MS 194.3 [M-56+H] ⁺ .

Synthesis 1981-C. To a solution of 1981-B (320 mg, 1.29 mmol) in DCM (6 mL) was added TFA (2 mL) dropwise. The reaction mixture was stirred at room temperature for 1 hour, after which the solution

- 11 043804 was concentrated in vacuo to give 1981-C as a crude product, which was used directly in the next step without further purification. MS 150.3 [M+H]+.

Synthesis 1981-D. A mixture of 1981-C (1.29 mmol, crude product from last step) and 1949-B (352 mg, 0.72 mmol) in DMSO (10 ml) was stirred at room temperature for 10 min, then Na ₂ CO ₃ ( 760 mg, 7.17 mmol) and the reaction mixture was stirred at room temperature for 2 hours. The mixture was then diluted with water (30 ml) and extracted with EtOAc (20 mlx3). The combined organic layers were washed with brine (20 mlx3), dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified by Prep-TLC (DCM:MeOH=40:1) to give 1981-D (205 mg, 67%) as a yellow oil. MS 427.1 [M+H] ⁺ .

Synthesis of Compound 7 A mixture of 1981-D (205 mg, 0.48 mmol) and Pd/C (205 mg) in MeOH/EtOAc (5/5 ml) was stirred at room temperature for 50 min under H ₂ atmosphere. Pd/C was removed by filtration through Celite, the filtrate was concentrated in vacuo and the residue was purified by Prep-TLC (DCM:MeOH=25:1) to give compound 7 (130 mg, 68%) as a brown solid. MS 397.2 [M+H] ⁺ .

Chiral resolution of compound 7. Enantiomers of racemic compound 7 (100 mg, 0.25 mmol) were separated by chiral chromatographic separation (Column: Chiralpak OJ-3; Solvent: MeOH; Flow rate: 2 ml/min; RT _8E i=2.287 min, RT _8E2 =2.553 min) to obtain the first elution peak of Enantiomer 1 (compound 8E1) (40 mg, 40%) as a yellow solid (MS 397.2 [M+H] ⁺ ) and the second elution peak of enantiomer 2 (compound 8E2) (20 mg, 20%) as a yellow solid. MS 397.2 [M+H] ⁺ . Stereochemistry was randomly assigned.

Compounds 9-21 were synthesized similarly using appropriately substituted versions of the boronic acid and brominated reagents used for the synthesis of compound 7.

Compound 9. 11 mg, 54%, yellow solid.

Compound 10. 11 mg, 47%, white solid.

Compound 11. 34 mg, 70%, white solid.

Compound 12. 18 mg, 48%, white solid.

Compound 13. 34 mg, 61%, light yellow solid.

Compound 15. 14 mg, 47%, white solid.

Compound 16. 35 mg, 63%, off-white solid.

Compound 17. 25 mg, 45%, white solid.

Compound 18. 30 mg, 52%, off-white solid.

Compound 19. 15 mg, 28%, light yellow solid.

Compound 20. 11 mg, 47%, light yellow solid.

Compound 21. 17 mg, 41%, light yellow solid.

Example 3. Synthesis of 14-E1 and 14-E2.

Synthesis 2147-A. To a mixture of tert-butyl 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,5-dihydro1H-pyrrole-1-carboxylate (33.6 g, 113 .9 mmol), 2-bromo-5-fluoropyrimidine (20 g, 113.6 mmol) and K2CO3

- 12 043804 (47.1 g, 341 mmol) in dioxane/H2O (500 ml/50 ml) PdCl ₂ (dppf) ₂ (4.6 g, 5.7 mmol) was added under nitrogen atmosphere. The resulting mixture was stirred at 100°C for 2 hours and then concentrated in vacuo. The residue was dissolved in EtOAc (500 ml) and the solution was washed with brine (200 mlx3). The organic layer was dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified by silica gel column chromatography. (PE:EAAc=10:1-5:1) to give 2147-A (25.5 g, 85%) as a gray solid. MS 210.1 [M-55] ⁺ .

Synthesis 2147-B. To a solution of 2147-A (1.0 g, 6.0 mmol) in DCM (30 mL) was added TFA (10 mL) dropwise at 0°C. The resulting solution was stirred at room temperature for 1 hour, after which the solvent was removed in vacuo to obtain 2147-B as a crude product, which was used directly in the next step. MS 166.1 [M+H] ⁺ .

Synthesis 2147-D. A solution of 1954-A (1.87 g, 7.45 mmol) in DMF (15 ml) was cooled to 0°C and then treated with NaH (60% in mineral oil) (596 mg, 14.9 mmol). The reaction mixture was stirred at 0°C for 30 minutes, then CDI (1.20 g, 7.45 mmol) was added and stirring was continued at 0°C for another 30 minutes. A solution of 2147-B in DMF was added to the reaction mixture and stirring was continued at 0°C for 1 hour. The mixture was then quenched with water (100 ml) and extracted with EtOAc (50 ml x 3). The combined organic layers were washed with brine (50 ml x 3), dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified by column chromatography (DCM:EtOAc=15:1-2:1) to give 2147-D (2.3 g, 70%) as a yellow solid. MS 443.2 [M+H] ⁺ .

Synthesis 14. A mixture of 2147-D (2.3 g, 5.2 mmol) and Pd/C (2.3 g) in MeOH/EtOAc (50 ml/50 ml) was stirred at room temperature for 1 h under H atmosphere ₂ . Pd/C was removed by filtration through a celite pad. The filtrate was concentrated in vacuo and the residue was purified by column chromatography (DCM:MeOH=20:1) to give compound 14 (1.14 g, 53%) as a white solid. MS 415.2 [M+H] ⁺ .

Chiral separation of 14-E1 and 14-E2. Enantiomers of 14 (1.14 g, 2.75 mmol) were separated using chiral SFC separation (column: Chiralcel OD-3; solvent: MeOH; flow rate: 2 ml/min; RT _t-2147-e1 = 1.849 min, RT _t-2147-E2 =2.175 min) to obtain compound 14-E1 isomer 1 (410 mg, 36%) as a yellow solid (MS 415.2 [M+H] ⁺ ) and compound 14-E2 (360 mg, 31%) as a yellow solid. MS 415.2 [M+H] ⁺ isomer 2.

Example 4. Synthesis of 19-E1 and 19-E2.

Synthesis 2145-A. A mixture of 1981-A (1.7 g, 6.8 mmol) and Pd/C (850 mg) in EtOAc (80 ml) was stirred at room temperature for 1 hour under H2. Pd/C was removed by filtration through celite. The filtrate was concentrated in vacuo to give 2145-A (1.6 g, 94%) as a colorless oil. MS 194.2 [M-55] ⁺ .

Synthesis 2145-B. To a solution of 2145-A (1.3 g, 522 mmol) in DCM (18 mL) was added TFA (6 mL) dropwise at 0°C. The reaction mixture was allowed to warm to room temperature and stirred at room temperature for 1 hour, after which the reaction mixture was concentrated in vacuo. The crude residue was dissolved in DMF (5 mL) and treated with TEA (1.58 g, 15.66 mmol) to give 2145-B as a solution used directly in the next step. MS 150.0 [M+H] ⁺ .

Synthesis of 2145-C Mixture of 6-chloro-3-nitropyridin-2-amine (50.0 g, 289.0 mmol), 4-fluorophenylboronic

- 13 043804 acid (48.5 g, 346.8 mmol) and K2CO ₃ (119.6 g, 867 mmol) in dioxane/H2O (1000/100 ml) were treated with Pd(PPh ₃ ) ₄ (5.0 g, 4.33 mmol) in an N2 atmosphere. The mixture was stirred at 95°C for 4 hours and then concentrated in vacuo. The residue was dissolved in EtOAc (2000 ml) and the solution was washed with brine (700 ml x 3). The organic layer was dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified by silica gel column chromatography (PE:EtOAc=10:1~5:1) to obtain 2145-C (36 g, 54%) as a yellow solid. MS 233.1 [M+H]+.

Synthesis 2145-E. A solution of 2145-C (1.0 g, 4.35 mmol) in DMF (20 ml) was cooled to 0°C and treated with NaH (60% in mineral oil) (210 mg, 5.22 mmol). The reaction mixture was stirred at 0°C for 30 minutes, then CDI (846 mg, 5.22 mmol) was added to the above mixture and stirring was continued at 0°C for another 30 minutes to obtain a solution as 2145-D. Solution 2145-B was added to solution 2145-D at 0°C and stirred for 1 hour. The reaction mixture was poured into water (420 ml), then extracted with EtOAc (50 ml x 3) and washed with brine (50 ml x 3). The organic layer was dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified by silica gel column chromatography (DCM:MeOH=100:1-30:1) to give 2145-E (1.2 g, 56.3%) as a yellow oil. MS 409.1 [M+H] ⁺ .

Synthesis 19. A mixture of 2145-E (1.2 g, 2.94 mmol) and Pd/C (1.2 g) in MeOH/EtOAc (20 ml/20 ml) was stirred at room temperature for 1 hour under H2 atmosphere. . Pd/C was removed by filtration through celite. The filtrate was concentrated in vacuo and the residue was purified by silica gel column chromatography (DCM:MeOH=100:1~15:1) to give 19 (700 mg, 63%) as an off-white solid. MS 379.4 [M+H] ⁺ .

Chiral separation of 19-E1 and 19-E2. Enantiomers of 19 (700 mg, 1.85 mmol) were separated using chiral SFC (column: Chiralpak AD-3; solvent: MeOH; flow rate: 1.5 ml/min; R ^T T-2145-E2= ^5.544 min to give 19-E2 (230 mg, 32.8%) as a white solid and RT _t-2145-e1 =4.009 min to give 19-E1 (260 mg, 37.1%) as a white solid.MS 379.4 [M+H] ⁺ .

Example 5: Alternative synthesis of 19-E1 and 19-E2.

Synthesis 2145-F. A solution of 2145-C (5.83 g, 25.0 mmol) in DMF (100 ml) was cooled to 0°C and treated with NaH (60% in mineral oil) (1.4 g, 35 mmol). The reaction mixture was stirred at 0°C for 30 minutes, then CDI (4.86 g, 30 mmol) was added to the above mixture and stirring was continued at 0°C for another 30 minutes to obtain solution 2145-D. A solution of 1981-B (9.26 g, 37.5 mmol) and TEA (18.93 g, 187.5 mmol) in DMF (40 mL) was then added to the 2145-D solution at 0°C and the resulting reaction mixture was stirred at 0°C for 1 hour. The reaction mixture was then poured into water (420 ml) and stirred for 10 minutes. The precipitate was collected by filtration and the precipitate was washed with water (150 ml), then with acetone (150 ml). Finally, the residue was concentrated to dryness to give 2145-F (9.5 g, 94%) as a light yellow solid. MS 407.1 [M+H] ⁺ .

Synthesis 19. A mixture of 2145-F (8.5 g, 20.9 mmol) and Pd/C (8.5 g) in MeOH/DCM (250 ml/200 ml) was stirred at room temperature for 3 hours under an atmosphere of H ₂ . Pd/C was removed by filtration through celite. The filtrate was concentrated in vacuo and the residue was purified by silica gel column chromatography (DCM:MeOH=100:1~15:1) to give 19 (4.1 g, 50%) as a light yellow solid. MS 379.4 [M+H] ⁺ .

Compounds 46, 50, and 51 were synthesized analogously to 19 using respectively substituted boronic acid and aryl bromide reagents.

Compound 46. 100 mg, 63%, light yellow substance.

Chiral resolution of 50 and 51. Enantiomers 19 were separated using chiral SFC (column: Chiralpak AD-3; solvent: MeOH; flow rate: 1.5 ml/min; RT5o = 3.267 min and RT5 ₀ = 5.375 min.

- 14 043804

Compound 50. 200 mg, 29%, white solid.

Compound 51. 230 mg, 33%, white solid.

Example 6. Synthesis of 20-E1 and 20-E2.

Synthesis 2292-A. Mixture of 2-bromo-5-fluoropyrimidine (20 g, 113.6 mmol), tert-butyl-3-(4,4,5,5tetramethyl-1,3,2-dioxaborolan-2-yl)-2,5 - Dihydro-III-pyrrole-1-carboxylate (33.7 g, 113.6 mmol) and K ₂ CO ₃ (47.0 g, 340.8 mmol) in dioxane/H ₂ O (500 ml/50 ml) were treated Pd(dppf) ₂ Cl ₂ (4.64 g, 5.7 mmol) under N ₂ atmosphere. The reaction mixture was stirred at 90°C for 3 hours and then concentrated in vacuo. The residue was dissolved in EtOAc (200 ml) and the solution was washed with saline (100 mx3). The organic layer was dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified by silica gel column chromatography (PE:DCM=1:1 to DCM) to give 2292-A (25.5 g, 85%) as a gray solid. MS 210.1 [M-55] ⁺ .

Synthesis 2292-B. A mixture of 2292-A (11.7 g, 44.1 mmol) in DCM (100 ml) was treated with TLA (30 ml) at 0°C. The reaction mixture was allowed to warm to room temperature and stirred at room temperature for 1 hour. The solution was then concentrated in vacuo and the residue was dissolved in DMF (50 ml) and treated with TEA (13.4 g, 132.3 mmol) to give 2292- B as the solution used directly in the next step. MS 165.1 [M+H] ⁺ .

Synthesis of 2292-C. A solution of 2145-C (9.32 g, 40.0 mmol) in DMF (100 ml) was cooled to 0°C and then treated with NaH (60% in mineral oil) (1.92 g, 48.0 mmol). The reaction mixture was stirred at 0°C for 30 minutes, then CDI (7.13 g, 44.0 mmol) was added to the above mixture and stirring was continued at 0°C for another 30 minutes to obtain solution 2145-D. The 2292-B solution was then added to the above mixture at 0°C and the resulting mixture was stirred at 0°C for 1 hour. The reaction mixture was then poured into water (450 ml) and stirred for 10 minutes. The precipitate was collected by filtration and the filter cake was washed with water (150 ml), then with acetone (150 ml). Finally, the residue was concentrated to dryness to give 2292-C (12.2 g, 70%) as a light yellow solid. MS 424.9 [M+H] ⁺ .

Synthesis 20. A solution of 2292-C (12.2 g, 28.6 mmol) and Pd/C (12.2 g) in MeOH/DCM (400 ml/300 ml) was stirred at room temperature for 2.5 h in _H2 atmosphere. Pd/C was removed by filtration through celite. The filtrate was concentrated in vacuo and the residue was purified by silica gel column chromatography (DCM-DCM:EA=1:1) to give 20 (6.8 g, 60%) as a yellow solid. MS 397.0 [M+H] ⁺ .

Chiral separation of 20-E1 and 20-E2. Enantiomers of 20 (360 mg, 0.91 mmol) were separated using chiral SFC (column: Chiralcel OJ-3; solvent: MeOH; flow rate: 1.5 ml/min; RT _T-2292-E2 = 2.862 min to give 20 -E2 (100 mg, 28%) as a white solid and RT _T-2292-E1 = 2.338 min to give 20-E1 (88 mg, 25%) as a white solid MS 397.0 [M+H ] ⁺ .

- 15 043804

Example 7.

N-N

2007-A 2007-B

Synthesis 2007-A. A solution of tert-butyl-3-hydroxypyrrolidine-1-carboxylate (821 mg, 4.75 mmol) in THF (10 ml) was treated with 3-bromopyridazine (500 mg, 3.16 mmol) and KOH (798 mg, 4.25 mmol ) at room temperature. The reaction mixture was then heated to 70°C and stirred at 70°C for 16 hours. The mixture was then diluted with water (20 ml) and extracted with EtOAc (20 mlx3). The combined organic layers were washed with brine (20 mlx3), dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified by Prep-TLC (E1OAc:PE=10:1) to give 2007-A (70 mg, 8%) as a yellow oil. MS 288.2 [M+H]+.

Synthesis 2007-B. To a solution of 2007-A (70 mg, 0.26 mmol) in DCM (6 mL) was added TFA (2 mL) dropwise. The reaction mixture was stirred at room temperature for 1 hour, after which the solvent was removed in vacuo to obtain 2007-B as a crude product, which was used directly in the next step without further purification.

Synthesis 2007-C. A mixture of 1949-B (71.0 mg, 0.14 mmol) and 2007-B (0.26 mmol, crude product from last step) in DMSO (5 ml) was stirred at room temperature for 10 min, then treated with Na ₂ CO ₃ (276 mg, 2.6 mmol). The resulting reaction mixture was stirred at room temperature for 2 hours, after which the mixture was diluted with water (20 ml) and extracted with EtOAc (10 ml x 3). The combined organic layer was washed with brine (10 mlx3), dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified by Prep-TLC (EtOAc:PE=5:1) to give 2007-C (40 mg, 45%) as a yellow solid. MS 443.2 [M+H] ⁺ .

Synthesis 22. A mixture of 2007-C (40 mg, 0.09 mmol) in MeOH (6 ml) was treated with Raney Ni (20 mg) and stirred at room temperature for 30 min under H ₂ atmosphere. Raney Ni was then removed by filtration through Celite, the filtrate was concentrated and the residue was purified by Prep-TLC (DCM:MeOH=10:1) to give 22 (18 mg, 48%) as a pink solid. MS 413.2 [M+H] ⁺ .

Example 8.

2008-A 2008-B

Synthesis 2008-A. A solution of tert-butyl-3-hydroxypyrrolidine-1-carboxylate (375 mg, 3.32 mmol) in THF (10 ml) was treated with 2-bromopyrimidine (350 mg, 3.22 mmol) and t-BuOK (1.08 g, 9.66 mmol) at room temperature. The reaction mixture was then heated to 70°C and stirred at 70°C for 3 hours. The mixture was then diluted with water (30 ml) and extracted with EtOAc (20 mlx3). The combined organic layers were washed with brine (20 mlx3), dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified by Prep-TLC (E1OAc:PE=10:1) to give 2008-A (400 mg, 48%) as a colorless oil. MS 288.2 [M+H]+.

- 16 043804

Synthesis 2008-B. To a solution of 2008-A (400 mg, 1.51 mmol) in DCM (6 mL) was added TFA (2 mL) dropwise. The resulting reaction mixture was stirred at room temperature for 1 hour, after which the solvent was removed in vacuo to obtain 2008-B as a crude product, which was directly used in the next step without further purification. MS 188.2 [M+H]+.

Synthesis 2008-C. A mixture of 1949-B (370.0 mg, 0.76 mmol) and 2008-B (1.51 mmol, crude product from last step) in DMSO (10 ml) was stirred at room temperature for 10 min, then treated with Na ₂ CO ₃ (800 mg, 7.55 mmol). The resulting reaction mixture was stirred at room temperature for 2 hours, after which the mixture was diluted with water (30 ml) and extracted with EtOAc (30 ml x 3). The combined organic layers were washed with brine (30 mlx3), dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified with Prep-TLC (EAAc:PE=5:1) to give 2008-C (250 mg, 75%) as a yellow solid. MS 443.2 [M+H]+.

Synthesis 23. A mixture of 2008-C (200 mg, 0.45 mmol) in MeOH (8 ml) was treated with Pd/C (200 mg) and the reaction mixture was stirred at room temperature for 1 hour under H ₂ atmosphere. Pd/C was then removed by filtration through Celite, the filtrate was concentrated and the residue was purified by Prep-TLC (DCM:MeOH=10:1) to give 23 (84 mg, 42%) as MS 413.2 [M+H] ⁺ .

Example 9.

off-white solid.

25E1

25E2 chiral separation

Synthesis 2058-A. A mixture of zinc dust (896 mg, 13.8 mmol) and anhydrous DMA (3 ml) was treated with TMSC1 and 1,2-dibromoethane (0.24 ml, v/v = 7/5) and the resulting reaction mixture was stirred at room temperature for 20 minutes in an atmosphere of N ₂ . A solution of tert-butyl-3(iodomethyl)pyrrolidine-1-carboxylate (3.3 g, 10.6 mmol) in anhydrous DMA (4 ml) was then added and the resulting reaction mixture was stirred at room temperature for 16 hours under N ₂ . The mixture was then used directly in the next step as 2058-A. The concentration of 2058-A was approximately 1.0 mol/L in DMA.

Synthesis 2058-B. A mixture of 2-bromopyrimidine (734 mg, 4.61 mmol), CuI (87 mg, 0.46 mmol) and Pd(PPh ₃ ) ₄ (266 mg, 0.23 mmol) in anhydrous DMA (15 ml) under N atmosphere ₂ were treated with 2058-A (6.0 ml). The resulting mixture was stirred at 60°C for 48 hours under N2 atmosphere. The mixture was then diluted with water (50 ml) and extracted with EtOAc (30 mlx3). The combined organic layers were washed with brine (30 mlx3), dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified by Prep-TLC (EtOAc:DCM=1:1) to give 2058-B (500 mg, 41%) as a yellow solid. MS 264.2 [M+H] ⁺ .

Synthesis 2058-C. To a solution of 2058-B (500 mg, 1.9 mmol) in DCM (10 mL) was added TFA (3 mL) dropwise. The reaction mixture was stirred at room temperature for 1 hour, after which the solution was concentrated in vacuo to give 2058-C as a crude product, which was used directly in the next step without further purification. MS 164.2 [M+H] ⁺ .

Synthesis 2058-D. A mixture of 2058-C (1.9 mmol, crude product from last step) and 1949-B (518 mg, 1.05 mmol) in DMSO (15 ml) was stirred at room temperature for 10 min, then treated with Na ₂ CO ₃ (1.11 g, 10.5 mmol) and the resulting reaction mixture was stirred at room temperature for 2 hours. The mixture was then diluted with water (50 ml) and extracted with EtOAc (30 ml x 3).

The combined organic layers were washed with brine (30 mlx3), dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified with Prep-TCS (DCM:EtOAc=1:2) to give 2058-D (400 mg, 86%) as a yellow solid. MS 441.2 [M+H]+.

Synthesis 24 A mixture of 2058-D (400 mg, 0.91 mmol) and Pd/C (400 mg) in MeOH/EtOAc (10 ml/10 ml) was stirred at room temperature for 1 hour under H ₂ atmosphere. Pd/C was removed by filtration through Celite, the filtrate was concentrated in vacuo and the residue was purified by Prep-TLC (DCM:MeOH=25:1) to give 24 (250 mg, 67%) as a yellow solid. MS 411.2 [M+H] ⁺ .

Chiral Resolution 24 Enantiomers 24 (250 mg, 0.61 mmol) were separated using chiral chromatographic separation (column: Chiralpak AD-3; solvent: MeOH; flow rate: 2 ml/min; RT _24E1 = 2.893 min, RT _24E2 = 3.892 min ) to yield enantiomer 1 (25E1) (62 mg, 25%) as a yellow solid (MC 411.2 [M+H] ⁺ ) and enantiomer 2 (25E2) (90 mg, 36%) as a yellow solid . MS 411.2 [M+H] ⁺ . Stereochemistry was randomly assigned.

Compounds 26-28 were synthesized similarly using suitably substituted brominated versions of the reagent used for the synthesis of 23.

Compound 26. 20 mg, 21%, yellow solid.

Compound 27. 110 mg, 59%, white solid.

Compound 28. 20 mg, 54%, light yellow solid.

Example 10.

Synthesis 2106-A. To a solution of 2-bromopyrimidine (50.0 g, 314.5 mmol) in DCM (600 ml) under nitrogen atmosphere, n-BuLi (150 ml, 377.5 mmol) was added dropwise at minus 78°C and stirred at minus 78 °C for 2 hours under nitrogen atmosphere. Tert-butyl 3-oxopyrrolidine-1-carboxylate (70 g, 377.5 mmol) in DCM (200 ml) was then added dropwise to the above mixture at minus 78°C. The resulting mixture was warmed to room temperature for 3 hours. The mixture was quenched with saturated NH4Cl (200 ml) and extracted with DCM (400 mlx3). The combined organic layers were washed with brine (200 mlx3), dried over anhydrous Na2SO4 and then concentrated in vacuo. The residue was purified by silica gel column chromatography (PE:EtOAc=10:1 to EtOAc) to give 2106-A (9.0 g, 11%) as a light yellow solid. MS 266.2 [M+H] ⁺ .

Synthesis 2106-B. To a solution of 2106-A (9.0 g, 34.0 mmol) in DCM (50 mL), DAST (18 mL) was added dropwise at −78°C and the solution was warmed to room temperature for 1 h under nitrogen atmosphere. The solvent was concentrated and the residue was purified by silica gel column chromatography (PE:EtOAc=10:1 to EtOAc) to give 2106-B (2.2 g, 24%) as a brown solid. MS 268.2 [M+H] ⁺ .

Synthesis 2106-C. To a solution of 2106-B (2.2 g, 8.21 mmol) in DCM (20 mL) 8 mL of TFA was added dropwise at 0°C. The solution was then stirred at room temperature for 1 hour. The solvent was removed in vacuo to give 2106-C as a crude product, which was used directly in the next step. MS 168.2 [M+H] ⁺ .

Synthesis 2106-D. A mixture of 1949-B (2.6 g, 5.47 mmol) and 2106-C (8.21 mmol, crude product from last step) in DMSO (40 ml) was stirred at room temperature for 10 min, followed by Na ₂ CO ₃ (5.8 g, 54.7 mmol) was added to the above mixture. The resulting mixture was continued to stir at room temperature for 2 hours. The mixture was diluted with water (200 ml), extracted with EtOAc (100 ml x 3). The combined organic layers were washed with brine (50 mlx3), dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified by silica gel column chromatography (PE:EtOAc=10:1 to EtOAc) to give 2106-D (2.0 g, 82%) as a yellow solid. MS 445.0 [M+H] ⁺ .

Synthesis 29 A mixture of 2106-D (12.0 g, 27.0 mmol) and Raney Ni (2.0 g) in MeOH (20 ml) was stirred at room temperature for 1 hour under H ₂ atmosphere. Raney's Ni was removed by filtration through a celite pad. The filtrate was concentrated in vacuo and the residue was purified by silica column chromatography

- 18 043804 kagel (PE:EtOAc=10:1 to EtOAc) to give 29 (8.0 g, 71%) as a yellow solid. MS 415.2 [M+H]+.

Chiral separation of 29-E1 and 29-E2. The enantiomers of compound 29 (8.0 g, 19.3 mmol) were separated using chiral SFC (column: Chiralcel OX-3; solvent: MeOH; flow rate: 1.5 ml/min; RT2i06 _-E i=2.814 min, RT _T-2 i06 _-E2 =4.362 min) to give 29-E1 (1.2 g, 11%) as a yellow solid (MS 415.2 [M+H]+) and 29-E2 (1.3 g , 12%) as a yellow solid. MS 415.2 [M+H] ⁺ .

Compound 45 was synthesized analogously to 29 using 2-(5-fluorothiophen-2-yl)-4,4,5,5tetramethyl-1,3,2-dioxoborolane as a reagent.

Compound 45. 90 mg, 19%, yellow solid.

Example 11.

Synthesis 1984-A. A solution of 2-bromo-5-methylthiazole (1.0 g, 5.59 mmol) in THF (20 ml) under N2 was treated dropwise with n-BuLi (2.7 ml, 6.70 mmol) at minus 78°C and the resulting reaction mixture was stirred at minus 78°C for 1 hour. Then a solution of tert-butyl 3-oxopyrrolidine-1-carboxylate (1.2 g, 6.70 mmol) was added dropwise to the reaction mixture at minus 78°C. in THF (10 ml). The reaction mixture was then allowed to warm to room temperature and stirred at room temperature for 3 hours. The mixture was diluted with saturated aqueous NH4Cl (40 ml) and extracted with EtOAc (30 ml x 3 ). The combined organic layers were washed with brine (20 mlx3), dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified by silica gel column chromatography (PE:EtOAc=10:1 to EtOAc) to give 1984-A (760 mg, 48%) as a light yellow solid. MS 285.2 [M+H]+.

Synthesis 1984-B. A solution of 1984-A (660 mg, 2.32 mmol) in DCM (10 ml) was cooled to 0°C and treated with pyridine (1.09 g, 13.94 mmol) followed by dropwise addition of SOCl ₂ (414 mg, 3 .48 mmol). The resulting reaction mixture was then heated to 45°C and stirred at 45°C for 16 hours. The mixture was then diluted with water (20 ml) and extracted with DCM (30 mlx3). The combined organic layers were washed with brine (20 mlx3), dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified by silica gel column chromatography (PE:EtOAc=10:1~1:5) to give 1984-B (120 mg, 19%) as a brown oil. MS 266.2 [M+H]+.

Synthesis 1984-C. A mixture of 1984-B (100 mg, 0.38 mmol) and Pd/C (100 mg) in MeOH (6 ml) was stirred at room temperature for 1 h under H2. Pd/C was then removed by filtration through Celite, the filtrate was concentrated and the residue was purified by Prep-TLC (EtOAc:PE=5:1) to give 1984-C (90 mg, 88%) as a light brown oil. MS 269.2 [M+H]+.

Synthesis 1984-D. To a solution of 1984-C (90 mg, 0.34 mmol) in DCM (3 mL) was added TFA (1 mL) dropwise. The resulting reaction mixture was stirred at room temperature for 1 hour, after which the solvent was removed in vacuo to obtain 1984-D as a crude product, which was used directly in the next step without further purification. MS 169.2 [M+H]+.

Synthesis 1984-E. A mixture of 1949-B (93 mg, 0.19 mmol) and 1984-D (0.34 mmol, crude from last step) in DMSO (5 mL) was treated with Na ₂ CO ₃ (200 mg, 1.89 mmol) and the resulting reaction mixture was stirred at room temperature for 2 hours. The mixture was then diluted with water (20 ml) and extracted with EtOAc (20 mlx3). The combined organic layers were washed with brine (20 mlx3), dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified by Prep-TLC (EtOAc:PE=5:1) to give 1984-E (80 mg, 95%) as a yellow solid. MS 446.2 [M+H] ⁺ .

Synthesis 30. A mixture of 1984-E (80 mg, 0.18 mmol) and Pd/C (80 mg) in MeOH (5 ml) was stirred at room temperature for 1 hour under H ₂ atmosphere. Pd/C was then removed by filtration through Celite, the filtrate was concentrated and the residue was purified by Prep-TLC (EtOAc:MeOH=15:1) to give 30 (44 mg, 59%) as a yellow solid. MS 416.2 [M+H]+.

- 19 043804

Example 12. Synthesis 31.

1954-A 1954-B

1954-C 1954-D

Synthesis 1954-A. A mixture of 6-chloro-3-nitropyridin-2-amine (4.58 g, 26.4 mmol), 2,4-difluorophenylboronic acid (5.00 g, 31.7 mmol) and K ₂ CO ₃ (10.9 g, 79.2 mmol) in dioxane/H ₂ O (100 ml/10 ml) was treated with Pd(PPh ₃ ) ₄ (1.10 g, 0.95 mmol) under nitrogen. The mixture was stirred at 100°C for 3 hours and then concentrated in vacuo. The residue was dissolved in EtOAc (200 ml) and the solution was washed with saline (100 ml x 3). The organic layer was dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified by silica gel column chromatography (PE:EtOAc=7:1-5:1) to give 1954-A (4.0 g, 61%) as a yellow solid. MS 252.1 [M+H]+.

Synthesis 1954-B. To a stirred solution of 1954-A (4.0 g, 15.94 mmol) in pyridine (60 mL) was added phenylcarbonochloridate (7.50 g, 47.81 mmol) dropwise at 0°C. After addition was complete, the mixture was stirred at 50°C for 4 hours. The mixture was concentrated in vacuo. The residue was purified by silica gel column chromatography (PE:DCM=3:2-1:1) to give 1954-B (7.1 g, 91%) as a yellow solid. MS 492.1 [M+H] ⁺ .

Synthesis 1954-C. To a mixture of zinc dust (449 mg, 6.9 mmol) in anhydrous DMA (2 ml) was added TMSC1 and 1,2-dibromoethane (0.24 ml, v/v = 7/5) and the reaction mixture was stirred at room temperature for 20 minutes in a nitrogen atmosphere. Then, a solution of tert-butyl-3-(iodomethyl)pyrrolidine-1-carboxylate (1.65 g, 5.3 mmol) in anhydrous DMA (1.5 ml) was added to the above mixture, and the resulting mixture was stirred at room temperature for 16 hours under nitrogen atmosphere conditions. The mixture was used directly in the next step as 1954-C. The concentration of 1954-C was approximately 1.0 mol/L in DMA.

Synthesis 1954-D. 1954 was added to a mixture of 3-bromopyrimidine (243 mg, 1.54 mmol), CuI (30 mg, 0.15 mmol) and Pd(PPh3)4 (89 mg, 0.077 mmol) in anhydrous DMA (6 ml) under nitrogen atmosphere. -C (2.0 ml). The resulting mixture was stirred at 60°C for 72 hours under a nitrogen atmosphere. The mixture was then diluted with water (20 ml) and extracted with EtOAc (20 mlx3). The combined organic layers were washed with brine (20 mlx3), dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified by Prep-TLC (DCM:EtOAc=2:1) to give 1954-D (180 mg, 44%) as a yellow solid. MS 263.2 [M+H] ⁺ .

Synthesis 1954-E. To a solution of 1954-D (160 mg, 0.69 mmol) in DCM (6 mL) was added TFA (2 mL) dropwise at 0°C. The resulting reaction mixture was stirred at room temperature for 1 hour, then concentrated in vacuo to give 1954-E as a crude product, which was used directly in the next step. MS 163.2 [M+H] ⁺ .

Synthesis 1954-F. A mixture of 1954-E (0.69 mmol, crude product from the previous step) and 1954-B (188 mg, 0.38 mmol) in DMSO (6 ml) was stirred at room temperature for 10 min, then Na ₂ CO ₃ ( 403 mg, 3.8 mmol) was added to the above mixture and the resulting reaction mixture was stirred at room temperature for 2 hours. The mixture was then diluted with water (30 ml) and extracted with EtOAc (10 mlx3). The combined organic layers were washed with brine (10 mlx3), dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified by Prep-TLC. (DCM:EtOAc=1:2) to give 1954-F (110 mg, 66%) as a yellow solid. MS 440.1 [M+H] ⁺ .

- 20 043804

Synthesis of Compound 31 A mixture of 1954-F (110 mg, 0.25 mmol) and Pd/C (110 mg) in MeOH/EtOAc (5/5 ml) was stirred at room temperature for 50 min under H ₂ atmosphere. Pd/C was removed by filtration through a celite pad. The filtrate was concentrated in vacuo and the residue was purified by Prep-TLC (DCM:MeOH=30:1) to give 31 (45 mg, 44%) as a yellow solid. MS 410.1 [M+H] ⁺ .

Compound 33 was synthesized similarly to 31 by using tert-butyl 3-iodopyrrolidone-1carboxylate and 2-bromo-5-methylpyrimidine as reagents.

Compound 33. 38 mg, 41%, light yellow solid.

Compound 48 was synthesized similarly to 31 by using tert-butyl 3-iodopyrrolidone-1carboxylate, 2-bromopyrmidine and 2-fluorophenylboronic acid as reagents.

Compound 48. 38 mg, 41%, light yellow solid.

Example 13. Synthesis of compound 32.

Synthesis 1985-A. Mixture of 2-bromo-5-methyl-1,3,4-thiadiazole (700 mg, 3.89 mmol), tert-butyl-3-(4,4,5,5tetramethyl-1,3,2-dioxaborolane-2 -yl)-2,5-dihydro-1H-pyrrole-1-carboxylate (1.15 g, 3.89 mmol) and Na ₂ CO ₃ (1.2 g, 11.7 mmol) in dioxane (40 ml) treated with PdCl ₂ (dppf) ₂ (159 mg, 0.2 mmol) under nitrogen. The reaction mixture was stirred at 90°C for 3 hours and then concentrated in vacuo. The residue was dissolved in EtOAc (30 ml) and the solution was washed with brine (10 ml x 3). The combined organic layer was dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified by silica gel column chromatography (PE:EAAc=10:1-1:1) to give 1985-A (400 mg, 39%) as a yellow solid. MS 268.1 [M+H]+.

Synthesis of 1985-B A mixture of 1985-A (400 mg, 1.5 mmol) and Pd/C (400 mg) in EtOAc (10 ml) was stirred at room temperature for 1 h under H2 atmosphere. The Pd/C was then removed by filtration through a celite pad. The filtrate was concentrated and the residue was purified by Prep-TLC (EA:PE=3:1) to give 1985-B (300 mg, 74%) as a yellow solid. MS 270.2 [M+H] ⁺ .

Synthesis 1985-C. A solution of 1985-B (300 mg, 1.1 mmol) in DCM (10 mL) was cooled to 0°C and then TFA (4 mL) was added dropwise at 0°C. The resulting solution was stirred at room temperature for 1 hour, after which the solvent was removed in vacuo to obtain 1985-C as a crude product, which was used directly in the next step. MS 170.2 [M+H] ⁺ .

Synthesis 1985-D. A mixture of 1954-B (300 mg, 0.6 mmol) and 1985-C (1.1 mmol, crude product from previous step) in DMSO (10 ml) was treated with Na ₂ CO ₃ (636 mg, 6.0 mmol), and the resulting mixture was stirred at room temperature for 2 hours. The mixture was then diluted with water (50 ml) and extracted with EtOAc (50 ml x 3). The combined organic layers were washed with brine (20 mlx3), dried over anhydrous Na2SO4 and then concentrated in vacuo. The residue was purified by Prep-TLC (EA:PE=5:1) to give 1985-D (150 mg, 56%) as a yellow solid. MS 447.2 [M+H] ⁺ .

Synthesis 32 A mixture of 1985-D (150 mg, 0.34 mmol) and Pd/C (150 mg) in MeOH (6 ml) was stirred at room temperature for 1 hour under H ₂ atmosphere. The Pd/C was then removed by filtration through a celite pad. The filtrate was concentrated and the residue was purified by Prep-TLC (EA:MeOH=15:1) to give 32 (83 mg, 55%) as a yellow solid. MS 417.2 [M+H] ⁺ .

Compound 37 was synthesized analogously to 32 using an appropriately substituted aryl bromide reagent.

Compound 37. 65 mg, 58%, light yellow solid.

- 21 043804

Example 14. Synthesis of compound 34.

^2060A 2060-B

Synthesis 2060-A. A solution of 1H-imidazole (115 mg, 1.69 mmol) in DMF (5 ml) was cooled to 0°C and treated with NaH (60% in mineral oil, 122 mg, 3.1 mmol). The reaction mixture was stirred at 0°C for 10 min, then tert-butyl 3-(iodomethyl)pyrrolidine-1-carboxylate (687 mg, 2.21 mmol) was added and the reaction mixture was heated to 40°C and stirred at 40°C for 3 hours. The mixture was then diluted with water (30 ml) and extracted with EtOAc (20 mlx3). The combined organic layers were washed with brine (20 mlx3), dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified by Prep-TLC (DCM:MeOH=30:1) to give 2060-A (105 mg, 25%) as a colorless oil. MS 197.2 [M+H] ⁺ .

Synthesis 2060-B. To a solution of 2060-A (201 mg, 0.80 mmol) in DCM (6 ml) TFA (2 ml) was added dropwise at 0°C. The resulting reaction mixture was allowed to warm to room temperature and stirred at room temperature for 1 hour, then concentrated in vacuo to give 2060-B as a crude product, which was used directly in the next step. MS 151.2 [M+H] ⁺ .

Synthesis of 2060-C A mixture of 2060-B (0.80 mmol, crude product from the previous step) and 1954-B (216 mg, 0.44 mmol) in DMSO (6 ml) was stirred at room temperature for 10 min, then Na ₂ CO ₃ (471 mg, 4.44 mmol) was added to the above mixture and stirred at room temperature for 2 hours. The mixture was diluted with water (20 ml) and extracted with EtOAc (10 mlx3). The combined organic layers were washed with brine (10 mlx3), dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified by Prep-TLC (DCM:MeOH=30:1) to give 2060-C (147 mg, 78%) as a yellow solid. MS 429.1 [M+H] ⁺ .

Synthesis 34. A mixture of 2060-C (124 mg, 0.29 mmol) and Pd/C (124 mg) in MeOH/EtOAc (5/5 ml) was stirred at room temperature for 2 hours under H ₂ atmosphere. Pd/C was removed by filtration through a celite pad. The filtrate was concentrated in vacuo and the residue was purified by Prep-TLC (DCM:MeOH=20:1) to give 34 (53 mg, 43%) as a white solid. MS 399.2. [M+N] ⁺ .

Compounds 35 were synthesized similarly to 34 using pyrazole as a reagent.

Compound 35. 60 mg, 64%, white solid. Example 16. Synthesis of compound 41. no ₂ '-A'. 'yS Pd(PPh ₃ ) ₄ . K ₂ CO ₃ , T 1 dioxane/H ₇ O A. ui ^ph vrS ---------- o pyridine T

Y O' Pd/C. N; EA street 2200-С About f 2200-F \ Р S 2200-А — 2200-В УУ ТФК/ДХМ 4 λ ^Ν 2200-В „ №\ Ν—\ , ) \ ) \ ChaoSOZDMSO ί^ΝΗΤΦΚ 2200-D 2200-E ^N Ni Raney, n _? H 7 ^Ng 0 MeOH

-22043804

Synthesis 2200-A. To a mixture of thiophen-2-ylboronic acid (14.1 g, 110 mmol), 6-chloro-3-nitropyridin2-amine (17.3 g, 100 mmol) and K ₂ CO ₃ (41.4 g, 300 mmol) in dioxane/H ₂ O (500/50 ml) Pd(PPh ₃ ) ₄ (5.8 g, 5.0 mmol) was added under nitrogen. The reaction mixture was stirred at 100°C for 2 hours and then concentrated in vacuo. The residue was dissolved in EtOAc (200 ml) and the solution was washed with saline (100 ml x 3). The organic layer was dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified by silica gel column chromatography (PE:EAAc=10:1-5:1) to give 2200-A (20.4 g, 84%) as a yellow solid. MS 222.0 [M+H]+.

Synthesis 2200-B. To a stirred solution of 2200-A (4.42 g, 20 mmol) in pyridine (80 mL) was added phenylcarbonochloridate (3.12 g, 60 mmol) dropwise at 0°C. After addition was complete, the mixture was stirred at 50°C for 4 hours. The mixture was then concentrated in vacuo and the residue was purified by silica gel column chromatography (PE:DCM=3:2-1:1) to give 2200-B (8.57 g, 93%) as a yellow solid. MS 462.1 [M+H] ⁺ .

Synthesis 2200-D. A mixture of 2200-C (108 mg, 0.44 mmol) and Pd/C (108 mg) in EtOAc (15 ml) was stirred at room temperature for 1 hour under H ₂ atmosphere. Pd/C was removed by filtration through a celite pad. The filtrate was concentrated in vacuo and the residue was purified by Prep-TLC (EAAc:PE=1:5) to give 41 (100 mg, 92%) as a white solid. MS 250.1 [M+H] ⁺ .

Synthesis 2200-E. To a solution of 2200-D (100 mg, 0.40 mmol) in DCM (3 mL) TFA (1 mL) was added dropwise at °C. The resulting solution was stirred at room temperature for 1 hour and then the solvent was removed in vacuo to give 2200-E as a crude product, which was used directly in the next step. MS 194.1 [M+H] ⁺ .

Synthesis 2200-F. A mixture of 2200-E (0.4 mmol, crude product from the previous step) and 2200-B (103 mg, 0.22 mmol) in DMSO (6 ml) was stirred at room temperature for 10 min, then Na ₂ CO ₃ ( 234 mg, 2.2 mmol) and the reaction mixture was stirred at room temperature for 2 hours. The mixture was diluted with water (30 ml) and extracted with EtOAc (10 mlx3). The combined organic layers were washed with brine (10 mlx3), dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified by Prep-TLC (DCM:EtOAc=1:1) to give 2200-F (80 mg, 91%) as a yellow solid. MS 397.0 [M+H] ⁺ .

Synthesis 41 A mixture of 2200-F (80 mg, 0.20 mmol) and Raney Ni (80 mg) in MeOH (15 ml) was stirred at room temperature for 1 hour under H ₂ atmosphere. Raney's Ni was removed by filtration through a celite pad. The filtrate was concentrated in vacuo and the residue was purified by Prep-TLC (DCM:MeOH=20:1) to give 41 (43 mg, 58%) as a brown solid. MS 367.2 [M+H] ⁺

Compound 42 was synthesized analogously to 41 using methyl magnesium bromide and the correspondingly substituted boronic acid reagent.

Compound 42. 23 mg, 31%, yellow solid.

Example 17. Synthesis of 43 and 44.

Synthesis 2332-A. A mixture of 2147-A (1.0 g, 3.8 mmol) and Pd/C (1.0 g) in EtOAc (20 ml) was stirred at room temperature for 1 hour under H ₂ atmosphere. Pd/C was removed by filtration through a celite pad. The filtrate was concentrated in vacuo and the residue was purified by silica gel column chromatography (PE:EtOAc=5:1-3:1) to give 2332-A (1.0 g, 99%) as a white solid. MS 268.1 [M+H] ⁺ .

Synthesis 2332-B. A solution of 2332-A (1.0 g, 3.7 mmol) in DCM (21 ml) was cooled to 0°C and dropwise to

- 23 043804 TFA (7 ml) was added at 0°C. The reaction mixture was allowed to warm to room temperature and stirred at room temperature for 1 hour. The solvent was then removed in vacuo and the residue was dissolved in DMF (7 ml) and treated with TEA (1.01 g, 10 mmol) to give 2332-B in as the solution used directly for the next step. MS 168.1 [M+H]+.

Synthesis of 2332-C. To a solution of thiophene (20.0 g, 238 mmol) in THF (500 ml) under nitrogen atmosphere, n-BuLi (100 ml, 250 mmol) was added dropwise at minus 78°C and the reaction mixture was stirred at minus 78°C for 1 hour in a nitrogen atmosphere. N-fluorobenzenesulfonimide (78.8 g, 250 mmol) in THF (300 ml) was then added dropwise to the above mixture at minus 78°C and warmed to room temperature over 1 hour. The reaction mixture was then cooled to minus 78°C and Another portion of n-BuLi (100 ml, 250 mmol) was added dropwise at minus 78°C and stirred at minus 78°C for 1 hour. Finally, 2-isopropoxy- 4,4,5,5-tetramethyl1,3,2-dioxaborolane (46.5 g, 250 mmol) in THF (200 ml). the reaction mixture was allowed to warm to room temperature and stirred for 16 hours. The mixture was poured into cooled saturated NH ₄ Cl (2000 ml), extracted with PE (400 ml x 3), and the combined organic layers were washed with brine (400 ml x 3), dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo to give 2332-C (32 g) as the crude product used directly in the next step. MS 229.0 [M+H] ⁺ .

Synthesis 2332-D. Mixture of 6-chloro-3-nitropyridin-2-amine (18.9 g, 109.6 mmol), 2332-C (30 g, crude from previous step) and K2CO3 (45.37 g, 328.8 mmol) in dioxane/H ₂ O (400/40 ml) was treated with Pd(PPh ₃ ) ₄ (2.0 g) under a nitrogen atmosphere. The mixture was stirred at 95°C for 3 hours and then concentrated in vacuo. The residue was dissolved in EtOAc (500 ml) and the solution was washed with brine (200 mlx3). The organic layer was dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified by silica gel column chromatography (PE:DCM=10:1-2:1) to give 2332-D (9.0 g, 34% (two steps)) as a yellow solid. MS 240.0 [M+H] ⁺ .

Synthesis 2332-F. A solution of 2332-D (820 mg, 3.43 mmol) in DMF (10 ml) was cooled to 0°C and treated with NaH (60% in mineral oil) (275 mg, 6.86 mmol). The reaction mixture was stirred at 0°C for 30 minutes, then CDI (556 mg, 3.43 mmol) was added to the above mixture and stirring was continued at 0°C for another 30 minutes. Finally, the 2332-B solution was added to the above mixture at 0°C and stirred for 1 hour. The mixture was then quenched with water (60 ml) and extracted with EtOAc (30 ml x 3 ). The combined organic layers were washed with brine (30 mlx3), dried over anhydrous Na2SO4 and then concentrated in vacuo. The residue was purified by silica gel column chromatography (DCM:EtOAc=15:1-2:1) to give 2332-F (1.18 g, 80%) as a yellow solid. MS 433.0 [M+H] ⁺ .

Synthesis of T-2332. A mixture of 2332-F (1.18 g, 2.7 mmol) and Raney Ni (1.2 g) in MeOH (20 ml) was stirred at room temperature for 1 hour under H ₂ atmosphere. Raney's Ni was removed by filtration through a pad of celite. The filtrate was concentrated in vacuo and the residue was purified by silica gel column chromatography (DCM:MeOH=50:1-20:1) to give T-2332 (850 mg, 77%) as a red solid. MS 403.0 [M+H] ⁺ .

Chiral resolution of 43 and 44. T-2332 (850 mg, 2.11 mmol) was resolved by chiral resolution (column: Chiralcel OJ-3; solvent: MeOH; flow rate: 2 ml/min; RT ₄₃ = 2.141 min, RT44 = 2.689 min) to give 43 (300 mg, 35%) as a light purple solid (MS 403.0 [M+H]+) and 44 (190 mg, 22%) as a white solid. MS 403.0 [M+H] ⁺ .

- 24 043804

Example 18. Synthesis of compounds 52 and 53. sk

Synthesis 2303-A. 2-Chloro-5-fluoropyrimidine (50 g, 378.0 mmol) was stirred in a solution of HBr in AcOH (33 wt%, 250 ml) at 40°C for 16 hours. The reaction mixture was then cooled to room temperature and the precipitate was collected filtrate. The filter cake was dissolved in EtOAc (500 ml) and made alkaline to pH 9 with saturated Na ₂ CO ₃ . The resulting mixture was extracted with EtOAc (500 mlx2). The combined organic layers were washed with brine (100 mlx3), dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. PE (20 mL) was added to the residue and the precipitate was collected by filtration, then dried in vacuo to give 2303-A (35.0 g, 53%) as a light brown solid. MS 177.2 [M+H]+.

Synthesis 2303-B. To a solution of 2303-A (5.0 g, 28.4 mmol) in DCM (70 mL), n-BuLi (13.6 mL, 34.1 mmol) was added dropwise at −78°C and the reaction mixture was stirred for 1 hour in N ₂ atmosphere. A solution of SM-A (6.3 g, 34.1 mmol) in DCM (20 ml) was then added dropwise to the mixture. The resulting mixture was warmed to room temperature and stirred for 3 hours. The mixture was then diluted with saturated NH4Cl (100 ml), extracted with DCM (100 mlx3). The combined organic layer was washed with brine (100 mlx3), dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified by silica gel column chromatography (PE:EtOAc=10:1 to EtOAc) to give 2303-B (550 mg) as crude product. The crude product was purified by Prep-HPLC to give 2303-B (177 mg, 2.2%) as a brown solid. MS 284.2 [M+H] ⁺ .

Synthesis 2303-C. A solution of 2303-B (1.6 g, 5.63 mmol) in DCM (50 ml) was treated dropwise with DAST (3.2 ml) at minus 78°C under N ₂ atmosphere. Then the solution was warmed to room temperature and stirred for 2 hours. Then the reaction was quenched with ice water and extracted with DCM (30 mlx3). The combined organic layer was washed with brine (100 mlx3), dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified by silica gel column chromatography. (PE:EtOAc=10:1~1:1) to give 2303-C (850 mg, 53%) as a brown solid. MS 286.2 [M+H] ⁺ .

Synthesis 2303-D. TFA (4 mL) was added dropwise to a solution of 2303-C (850 mg, 2.98 mmol) in DCM (30 mL) at 0°C. The solution was then allowed to warm to room temperature and stirred at room temperature for 1 hour. The solvent was removed in vacuo to give 2303-D as a crude product, which was used directly in the next step.

Synthesis 2303-E. To a solution of SM-B (1.2 g, 2.48 mmol) and 2303-D (1.1 g, crude product from last step) in DMSO (50 ml) was added Na ₂ CO ₃ (3.2 g, 29 .8 mmol). The mixture was stirred at room temperature for 2 hours. The mixture was diluted with water (200 ml) and extracted with EtOAc (100 ml x 3). The combined organic layers were washed with brine (100 mlx3), dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified by Prep-TLC (PE:EtOAc=1:5) to give 2303-E (680 mg, 61%) as a yellow solid. MS 445.2 [M+H] ⁺ .

Synthesis of 52 and 53. A mixture of 2303-E (680 mg, 1.53 mmol) and Raney Ni (680 mg) in MeOH (10 ml) was stirred at room temperature for 1 h under H ₂ atmosphere. Raney Ni was then removed by filtration

- 25 043804 via Celite. The filtrate was concentrated and the residue was purified by Prep-TLC (EA:MeOH=10:1) to give T-2303 (600 mg, 94%) as a yellow solid. Enantiomers were separated using chiral SFC (column: Chiralcel OJ-3; solvent: MeOH; flow rate = 1.5 ml/min; RT ₅₂ = 1.8 69 min, RT ₅₃ = 2.848 min) to give 52 (250 mg, 41%) as a yellow solid. MS 415.2 [M+H]+. 53 (240 mg, 40%) as a yellow solid. MS 415.2 [M+H] ⁺ .

Example 19. Synthesis of 54 and 55.

Synthesis 2294-A. To a solution of 3-bromopyridazine (50.0 g, 314.5 mmol) in DCM (400 ml), n-BuLi (2.5 M in hexane) (150 ml) was added dropwise at minus 78°C under N2 atmosphere and the reaction the mixture was stirred at minus 78°C for 1 hour. A solution of tert-butyl-3oxopyrrolidine-1-carboxylate (69.7 g, 377.0 mmol) in DCM (200 ml) was then added to the above mixture and the mixture was warmed to room temperature. temperature and stirred for 3 hours. The reaction mixture was poured into saturated NH ₄ Cl (300 ml), then extracted with DCM (400 ml x 3) and washed with brine (300 ml x 3). The organic layer was dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified by silica gel column chromatography (PE:EAAc=10:1 to EtOAc) to give 2294-A (7.0 g) as crude product. MS 266.2 [M+H] ⁺ .

Synthesis 2294-B. A solution of 2294-A (7.0 g, crude from last step) in DCM (100 mL) was cooled to −78°C and treated with DAST (3.0 mL) dropwise, after which the reaction mixture was allowed to warm to room temperature and stirred at room temperature for 2 hours. The mixture was then diluted with water (100 ml) and extracted with DCM (50 ml x 3). The combined organic layer was washed with brine (80 mlx3), dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified by silica gel column chromatography (EtOAc:PE=10:1~1:1) to give 2294-B (2.5 g, 3%) (two steps) as a brown solid. MS 268.2 [M+H] ⁺ .

Synthesis of 2294-C A solution of 2294-B (2.5 g, 9.4 mmol) in DCM (24 ml) was cooled to 0°C and treated with 8 ml TFA. After addition, the reaction mixture was allowed to warm to room temperature and stirred at room temperature for 1 hour. The solution was concentrated in vacuo to give 2294-C as a crude product, which was used directly in the next step. MS 168.2 [M+H] ⁺ .

Synthesis 2294-D. A solution of SM-A (2.5 g, 5.3 mmol) and 2294-C (9.4 mmol, crude from last step) in DMSO (50 mL) was treated with Na ₂ CO ₃ (5.5 g, 52. 2 mmol). The reaction mixture was stirred at room temperature for 2 hours. The mixture was then diluted with water (150 ml), extracted with EtOAc (100 ml x 3) and the combined organic layers were washed with brine (100 ml x 3), dried over anhydrous NaiSCL and then concentrated in vacuo. The residue was purified by silica gel column chromatography (EtOAc-EtOAc:MeOH=30:1) to give 2294-D (1.4 g, 82%) as a yellow solid. MS 427.2 [M+H] ⁺ .

Synthesis of T-2294. A mixture of 2294-D (1.4 g, 3.3 mmol) and Raney Ni (1.0 g) in DCM/MeOH (10 mL/10 mL) was stirred at room temperature for 1 h under H ₂ atmosphere. Raney Ni was then removed by filtration through celite. The filtrate was concentrated and the residue was purified by silica gel column chromatography (EtOAc-EtOAc:MeOH=10:1) to give T-2294 (800 mg, 61%) as a gray solid. MS 397.2 [M+H] ⁺ .

Chiral Resolution 55. T-2294 (800 mg, 2.02 mmol) was resolved with chiral SFC (column: Chiralcel OJ-3; solvent: MeOH; flow rate = 2 mL/min; RT ₅₅ = 2.599 min) to give 55 (226 mg, 17%) as a yellow solid, (RT ₅₄ =1.854 min) to give 54 (226 mg, 17%) as a yellow solid. MS 397.2 [M+H] ⁺ .

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Example 20. Synthesis of 39 and 40.

Synthesis 2201-A. To a solution of thiophene (20.0 g, 238 mmol) in THF (400 ml), n-BuLi (2.5 M in hexane) (100 ml) was added dropwise at minus 78°C and the reaction mixture was stirred at minus 78°C for 1 hour. Then a solution of NFSI (78.8 g, 250 mmol) in THF (400 ml) was added dropwise to the above solution at minus 78°C and the reaction mixture was warmed to room temperature and stirred for 1 hour. Then the reaction the mixture was again cooled to minus 78°C, another portion of n-BuLi (2.5 M in hexane) (100 ml) was added dropwise to the above mixture at minus 78°C and stirring was continued at minus 78°C for 1 Finally, a solution of 2-isopropoxy-4,4,5,5-tetramethyl-1,3,2dioxaborolane (46.5 g, 250 mmol) in THF (200 ml) was added dropwise to the above solution at minus 78°C . The reaction mixture was warmed to room temperature for 16 hours. The reaction mixture was poured into saturated NH ₄ Cl (1000 ml), then extracted with PE (300 ml x 3) and washed with saline (300 ml x 3). The organic layer was dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo to give 2201-A (32 g) as crude product. MS 147.1 [M-82] ⁺ .

Synthesis 2201-B. Mixture of 6-chloro-3-nitropyridin-2-amine (18.9 g, 109.6 mmol), 2201-A (32 g, crude from last step) and K2CO3 (45.4 g, 328.8 mmol) in dioxane/H ₂ O (400 ml/40 ml) was added Pd(PPh ₃ ) ₄ (2.0 g, 1.73 mmol) under N2 atmosphere. The mixture was stirred at 95°C for 4 hours and then concentrated in vacuo. The residue was dissolved in EtOAc (300 ml) and the solution was washed with saline (100 ml x 3). The organic layer was dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified by silica gel column chromatography (PE:DCM=10:1-2:1) to give 2201-B (9.0 g, 34%) as a yellow solid. MS 240.1 [M+H] ⁺ .

Synthesis 2201-C. A stirred solution of 2201-B (460 mg, 1.92 mmol) in pyridine (10 mL) was treated dropwise with phenylcarbonochloridate (900 mg, 5.77 mmol). After addition was complete, the mixture was stirred at 55°C for 2 hours. The mixture was concentrated in vacuo. The residue was purified by silica gel column chromatography. (PE:DCM=20:1~1:3) to give 2201-C (700 mg, 76%) as a yellow solid. MS 479.8 [M+H] ⁺ .

Synthesis 2201-D. Na ₂ CO 3 (233 mg, 2.2 mmol) was added to a solution of 2201-C (106 mg, 0.22 mmol) and ₂₁₄₅ -B (200 mg, 0.40 mmol) in DMSO (10 mL). The reaction mixture was stirred at room temperature for 2 hours. The mixture was diluted with water (30 ml), extracted with EtOAc (20 mlx3). The combined organic layers were washed with brine (20 mlx3), dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified by Prep-TLC (PE:EA=1:5) to give 2201-D (75 mg, 82%) as a yellow solid. MS 415.2 [M+H] ⁺ .

Synthesis of T-2201. A mixture of 2201-D (75 mg, 0.18 mmol) and Raney Ni (75 mg) in DCM/MeOH (3 mL/5 mL) was stirred at room temperature for 0.5 h. Raney Ni was removed by filtration through Celite. The filtrate was concentrated and the residue was purified by Prep-TLC (EA:MeOH=10:1) to give T-2201 (15 mg, 22%) as a gray solid. MS 385.2 [M+H] ⁺ .

Chiral resolution of 39 and 40. T-2201 (1.0 g, 2.6 mmol) was resolved by chiral resolution (column: Chiralcel OJ-3; solvent: MeOH; flow rate: 1.5 mL/min; RT ₃₉ = 2.225 min , RT ₄₀ =2.667 min) to give 39 (300 mg, 30%) as a brown solid (MS 385.0 [M+H] ⁺ ) and 40 (300 mg, 30%) as a purple solid. MS 385.0 [M+H] ⁺ .

Compound 49 was synthesized analogously to 39 and 40 using the respectively substituted reagents boronic acid and aryl bromide.

Compound 49. 30 mg, 23%, yellow solid.

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Example 22. Synthesis of 36.

247B-V 2066-A 2M6-V 2066-S

Synthesis 2066-A. A solution of 2475-B (400 mg, 1.7 mmol) in THF (10 mL) was treated dropwise with LDA (2.6 mL, 5.2 mmol) at −78°C under nitrogen and stirred for 1 h at − 78°C. Then, a solution of tert-butyl-3-oxoazetidine-1-carboxylate (414 mg, 2.2 mmol) in THF (5 ml) was added dropwise to the above mixture at minus 78°C, and then the reaction mixture was allowed to warm to room temperature and stirred for 16 hours. The mixture was diluted with saturated NH ₄ Cl (40 ml), extracted with EtOAc (30 mlx3). The combined organic layers were washed with brine (30 mlx3), dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified by silica gel column chromatography (PE:ESAAc=3:1-ESAAc) to give 2066-A (380 mg, 84%) as a colorless oil. MS 264.2 [M+H]+.

Synthesis 2066-B. A mixture of 2066-A (380 mg, 1.4 mmol) and Pd/C (380 mg) in EtOAc (10 ml) was stirred at room temperature for 1 hour under H2. The Pd/C was then removed by filtration through a celite pad. The filtrate was concentrated in vacuo and the residue was purified by silica gel column chromatography (PE:EtOAc=3:1-EtOAc) to give 2066-B (300 mg, 79%) as a colorless oil. MS 266.2 [M+H] ⁺ .

Synthesis of 2066-C. To a solution of 2066-B (150 mg, 0.57 mmol) in DCM (6 mL) was added TFA (2 mL) dropwise at 0°C. The reaction mixture was allowed to warm to room temperature and stirred at room temperature for 1 hour. The solution was concentrated in vacuo to give 2066-C as a crude product, which was used directly in the next step. MS 166.2 [M+H] ⁺ .

Synthesis 2066-D. A mixture of 2066-C (0.57 mmol, crude product from previous step) and 1954-B (154 mg, 0.32 mmol) in DMSO (6 ml) was stirred at room temperature for 10 min, then Na ₂ CO ₃ ( 339 mg, 3.2 mmol) and the reaction mixture was stirred at room temperature for 2 hours. The mixture was then diluted with water (20 ml) and extracted with EtOAc (10 mlx3). The combined organic layers were washed with brine (10 mlx3), dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified by Prep-TLC (EtOAc:MeOH=40:1) to give 2066-D (100 mg, 73%) as a yellow solid. MS 443.1 [M+H] ⁺ .

Synthesis 36 A mixture of 2066-D (100 mg, 0.23 mmol) and Pd/C (100 mg) in MeOH/EtOAc (5 ml/5 ml) was stirred at room temperature for 50 min under H ₂ atmosphere. Pd/C was removed by filtration through a celite pad. The filtrate was concentrated in vacuo and the residue was purified by Prep-TLC (DCM:MeOH=30:1) to give 36 (40 mg, 42%) as a yellow solid. MS 413.0 [M+H] ⁺ .

Example 23. Synthesis of compound 47.

F F

Synthesis 2341-A. To a solution of 2-(1-(tert-butoxycarbonyl)pyrrolidin-3-yl) acetic acid (1.15 g, 5.0 mmol), HOBt (810 mg, 6.0 mmol) and EDCI (1.44 g, 7.5 mmol) in DCM (20 ml) was added DIPEA (1.94 g, 15.0 mmol) and stirred at room temperature for 30 min under nitrogen. A solution of prop-2-yne-1-amine (413 mg, 7.5 mmol) in DCM (10 ml) and hemochloride was then added to the above mixture.

- 28 043804 The mixture was stirred at room temperature for 24 hours. The mixture was diluted with DCM (30 ml), washed with 0.5 N. HC1 (20 mlx2), saturated Na ₂ CO ₃ (20 mlx2) and saline (20 mlx2). The organic layer was dried over anhydrous Na ₂ SO ₄ and then concentrated in vacuo. The residue was purified by silica gel column chromatography (PE:EtOAc=10:1-2:1) to give 2341-A (1.0 g, 75%) as a colored oil. MS 211.0 [M-55] ⁺ .

Synthesis 2341-B. A solution of 2341-A (580 mg, 2.2 mmol) in acetonitrile (20 ml) was treated with gold trichloride (50 mg, 0.075 mmol) and the reaction mixture was stirred at 45°C for 72 h under nitrogen. The mixture was concentrated in vacuo. The residue was purified by silica gel column chromatography (PE EtOAc=10:1-1:1) to give 2341-B (380 mg, 66%) as a colorless oil. MS 267.0 [M+H] ⁺ .

Synthesis of 2341-C. A solution of 2341-B (380 mg, 1.4 mmol) in DCM (12 mL) was cooled to 0°C and then TFA (4 mL) was added dropwise. After addition, the reaction mixture was allowed to warm to room temperature and stirred at room temperature for 1 hour. The reaction mixture was then concentrated in vacuo to give 2341-C as a crude product. The residue was then dissolved in DMF (6 mL) and treated with TEA (424 mg, 4.2 mmol) to give 2341-C as a solution, which was used directly in the next step. MS 167.0 [M+H] ⁺ .

Synthesis 2341-D. A solution of 1954-A (252 mg, 1.0 mmol) in DMF (5 ml) was cooled to 0°C and treated with NaH (60% in mineral oil, 80 mg, 2.0 mmol). The reaction mixture was stirred at 0°C for 30 minutes, then CDI (162 mg, 1.0 mmol) was added and the reaction mixture was stirred at 0°C for an additional 30 minutes. Finally, the 2341-C solution was added to the above mixture at 0°C and the mixture was stirred at 0°C for 1 hour. The mixture was quenched with water (50 ml) and extracted with EtOAc (20 mlx3). The combined organic layers were washed with brine (20 mlx3), dried over anhydrous Na2SO4 and then concentrated in vacuo. The residue was purified by silica gel column chromatography (PE:EtOAc=10:1-1:2) to give 2341-D (280 mg, 63%) as a yellow solid. MS 444.1 [M+H] ⁺ .

Synthesis 47 A mixture of 2341-D (280 mg, 0.63 mmol) and Pd/C (280 mg) in MeOH/EtOAc (10 mL/10 mL) was stirred at room temperature for 1 h under H2 atmosphere. Pd/C was removed by filtration through a celite pad. The filtrate was concentrated in vacuo and the residue was purified by Prep-HPLC to give 47 (220 mg, 85%) as a light yellow solid. MS 414.2 [M+H] ⁺ .

Table 1

No. Structure MS calculation MS find Ή NMR data (400 MHz, DMSO-de) 1 0^ o=< ZI τι 395 396 5 8.54 (d, E=4.8Hz, 1H), 8.32 (s, 1H), 7.98 -7.92 (w, 1H), 7.78 - 7.76 (m, 1H) , 7.38 (d,J =7.6 Hz, 2H), 7.31 -7.26 (m, 2H), 7.187.13 (t, 2H), 5.23 (s, 2H), 3, 88 - 3.86 (t, 1H), 3.69 3.46 (t, 4H), 2.32 - 2.29 (t, 1H), 2.17-2.13 (t, 1H).

- 29 043804

2 z—' ZI '------( '------' hO P 395 396 δ 8.57 (d, J = 8.0 Hz, 1H), 8.47 (dd, 7 = 4.8, 1.2 Hz, 1H), 8.35 (s, 1H), 7.98 7 .92 (m, 1H), 7.76 (d, 7=8.0 Hz, 1H), 7.40 - 7.26 (m, 3H), 7.18-7.14 (m, 2H), 5.24 (s,2H), 3.96 - 3.91 (m, 1H), 3.70 - 3.66 (m, 1H), 3.53 - 3.33 (m, 3H), 2, 35 2.30 (m, 1H), 2.10-2.00 (m, 1H). 3 LL ,-2 rv^ 395 396 δ 8.53 - 8.51 (m, 2H), 8.36 (s, 1H), 7.98 - 7.91 (m, 1H), 7.40 - 7.35 (m, 3H), 7.32 7.26 (m, 1H), 7.18- 7.14 (m, 2H), 5.23 (s, 2H), 3.94 3.90 (m, 1H), 3.66 - 3.63 (m, 1H), 3.50 - 3.41 (m, 3 H), 2.33-2.31 (m, 1H), 2.05 1.98 (m, 1H). 4 I Ο Η ^vyS ο σ' F 394 395 δ 8.32 (s, 1H), 7.98 - 7.92 (m, 1H), 7.40 - 7.24 (m, 7H), 7.16-7.14 (m, 2H), 5 .23 (s, 2H), 3.93 - 3.89 (m, 1H), 3.69 - 3.65 (m, 1H), 3.49 - 3.21 (m, 3H), 2.29 2.27 (m, 1H), 2.04 - 1.99 (m, 1H).

- 30 043804

5 o~° ~ U n 7* w V F 411 412 δ 8.39 (s, 1H), 8.19 (dd, J= 5.2, 1.2 Hz, 1H), 7.94-7.90 (m, 1H), 7.75-7.71 (m, 1H), 7.40 - 7.39 (m, 1H), 7.32-7.18 (m, 1H), 7.167.02 (m, 2H), 7.01 -6.99 (m , 1H), 6.85 (d, J =8.4 Hz, 1H), 5.71 (s, 1H), 3.79 3.76 (m, 1H), 3.67-3.38 (m , 3H), 2.27 - 2.13 (m, 2H). 6 9 a°h ZI -Q-6-f 411 412 δ 8.39 (s, 1H), 8.20-8.18 (m, 1H), 7.97 - 7.90 (m, 1H), 7.75 - 7.71 (m, 1H), 7 .38 (dd,/= 8.0, 2.4 Hz, 1H), 7.31 -7.25 (m, 1H), 7.17-7.13 (m, 2H), 7.00 (dd ,/=6.0, 5.2 Hz, 1H), 6.85 (d,/= 8.0 Hz, 1H), 5.57 (s, 1H), 5.21 (s, 2H), 3 .79 - 3.53 (m, 4H), 2.27 2.07 (m, 2H), 7 Ό ^ν N— Ο Η ΐ ^Η2 ^VrS 0 F 396 397 6 8.80 (d,/= 4.8 Hz, 2H), 8.34 (s, 1H), 7.98 - 7.91 (m, 1H), 7.42 - 7.37 (m, 2H ), 7.32 -7.26 (m, 1H), 7.187.13 (m, 2H), 5.23 (s, 2H), 3.90 - 3.87 (m, 1H), 3.77 3 .75 (m, 2H), 3.64 - 3.59 (m, 1H), 3.55-3.51 (m, 1H), 2.36 - 2.33 (m, 1H), 2.27 - 2.22 (m, 1H).

- 31 043804

8E1 "ABOUT* N f R or S ο Η O V F 396 397 δ 8.79(t, J= 4.8 Hz, 2H), 8.33 (s, 1H), 7.98 - 7.91 (m, 1H), 7.42 - 7.37 (m, 2H ), 7.31 -7.25 (m, 1H), 7.177.13 (m, 2H), 5.22 (s, 2H), 3.90 - 3.87 (m, 1H), 3.76 3 .75 (m, 2H), 3.65 - 3.60 (m, 1H), 3.56 - 3.49 (m, 1H), 2.37 - 2.33 (m, 1H), 2.28 - 2.22 (m, 1H). 8E2 τι 396 397 δ 8.79(t, J= 4.8 Hz, 2H), 8.33 (s, 1H), 7.98 - 7.91 (m, 1H), 7.42 - 7.37 (m, 2H ), 7.31 -7.25 (m, 1H), 7.177.13 (m, 2H), 5.22 (s, 2H), 3.90 - 3.87 (m, 1H), 3.76 3 .75 (m, 2H), 3.65 - 3.60 (m, 1H), 3.56 - 3.49 (m, 1H), 2.37 - 2.33 (m, 1H), 2.27 - 2.22 (m, 1H). 9 /=Nw 7%H ^VyS about 7 F 396 397 δ 9.14-9.12 (m, 1H), 8.36 (s, 1H), 7.96 - 7.90 (m, 1H), 7.71 - 7.65 (m, 2H), 7 .37 (dd, 7=8.4, 2.4 Hz, 1H), 7.30 -7.24 (m, 1H), 7.167.11 (m, 2H), 5.22 (s, 2H), 3.97 - 3.93 (m, 1H), 3.80 3.76 (m, 1H), 3.68 - 3.65 (m, 2H), 3.55 - 3.49 (m, 1H) , 2.39-2.31 (m, 1H), 2.21-2.18 (m, 1H).

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10 LL τ τΓΑ Η ^Η y/vjH ΙΖ ΧΫ ³ 410 411 5 8.62 (d, 7=5.2 Hz, 2H), 8.32 (s, 1H), 7.98 - 7.91 (m, 1H), 7.31 - 7.28 (m, 2H ), 7.17-7.13 (m, 2H), 5.22 (s, 2H), 3.90 - 3.86 (m, 1H), 3.76 - 3.64 (m, 3H), 3.52 3.49 (m, 1H), 2.46 (s, 3H), 2.33 - 2.23 (m, 2H). eleven Ll ΣΖ 409 410 5 8.41 (d, 7 =2.4 Hz, 1H), 8.01 - 7.95 (m, 1H), 7.58 (dd, 7 = 8.0, 2.4 Hz, 1H), 7.45 (dd, 7=8.0, 4.8 Hz, 1H), 7.19-1.17 (m, 3H), 7.07 - 6.98 (m, 2H), 4.74 ( s, 2H), 3.95 - 3.91 (m, 1H), 3.73 - 3.69 (m, 1H), 3.56 3.38 (m, 3H), 2.47 (s, 3H ), 2.35 -2.34 (m, 1H), 2.181.93 (m, 1H). 12 3 Μ ΗΖ \=(VT> -Π 439 440 5 8.33 (s, 1H), 7.98 - 7.92 (m, 1H), 7.78 (t, 7= 7.6 Hz, 1H), 7.38 (dd, 7=8.0 , 2.0 Hz, 1H), 7.32 - 7.26 (m, 3H), 7.18-7.14 (m, 2H), 5.23 (s, 2H), 4.48 (s, 2H), 3.69 - 3.87 (m, 1H), 3.59 3.48 (m, 4H), 3.37 (s, 3H), 2.28 -2.27 (m, 1H), 2.162.11(m, 1H).

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13 A O n ^ ⁿ y ⁿ yS ο σ' F 424 425 δ 8.35 (s, 1H), 7.97-7.91 (m, 1H), 7.40 - 7.37 (m, 1H), 7.32-7.26 (m, 1H), 7 ,18-7.13 (m, 3H), 5.22 (s, 2H), 3.83 (t,7=7.6 Hz, 1H), 3.67 - 3.60 (m, 2H), 3.57 - 3.46 (m, 2H), 2.55 (s, 3H), 2.51 (s, 3H), 2.50 - 2.39 (m, 1H), 2.33 -2, 10 (m, 1H). 14 ZI P 414 415 5 8.89 (d, 7= 0.8 Hz, 2H), 8.35 (s, 1H), 7.97 - 7.91 (m, 1H), 7.40 - 7.37 (m, 1H ), 7.32 -7.26 (m, 1H), 7.187.13 (m, 2H), 5.22 (s, 2H), 3.94 - 3.88 (m, 1H), 3.80 3 .72 (m, 2H), 3.61 -3.59 (m, 1H), 3.55 - 3.52 (m, 1H), 2.38 -2.33 (m, 1H), 2.25 - 2.22 (m, 1H). 14E1 F _Z N $ ^H T ^v VyS o V F 414 415 5 8.88 (s, 2H), 8.54 (s, 1H), 7.97-7.91 (m, 1H), 7.38 (dd, 7=8.4, 2.4 Hz, 1H ), 7.31 7.25 (m, 1H), 7.18-7.13 (m, 2H), 5.22 (s, 2H), 3.923.88(m, 1H), 3.80- 3 .71 (m, 2H), 3.62-3.59 (m, 1H), 3.53-3.51 (m, 1H), 2.362.32 (m, 1H), 2.32 - 2.22 (m, 1H),

- 34 043804

14E2 IZ x/ ⁵ 414 415 δ 8.88 (s, 2H), 8.54 (s, 1H), 7.97 - 7.91 (m, 1H), 7.38 (dd, 7=8.4, 2.4 Hz, 1H ), 7.31 -7.25 (m, 1H), 7.187.13 (m, 2H), 5.22 (s,2H), 3.92 - 3.88(m, 1H), 3.80 3 .71 (m, 2H), 3.62 - 3.59 (m, 1H), 3.53-3.51 (m, 1H), 2.36 - 2.32 (m, 1H), 2.32 - 2.22 (m, 1H). 15 / ^N 4 w 7a h ^vrS 0 nA F 410 411 CD3CN as solvent, δ 8.56 (d, 7=5.2 Hz, 1H), 8.02 - 7.96 (m, 1H), 7.46 (dd, 7=8.0, 2.4 Hz , 1H), 7.19-7.14 (m, 3H), 7.07 6.98 (m, 2H), 4.73 (s, 2H), 3.91 - 3.87 (m, 1H) , 3.72 3.67 (m, 2H), 3.60 - 3.54 (m, 2H), 2.62 (s, 3H), 2.37 2.36 (m, 1H), 2.23 -2.15 (m, 1H). 16 LL ^xz >° A 424 425 6 8.33 (s, 1H), 7.98-7.91 (m, 1H), 7.38 (dd, 7= 8.0, 2.4 Hz, 1H), 7.32 - 7.26 (m, 1H), 7.17-7.14 (m, 3H), 5.22 (s, 2H), 3.88 - 3.84 (m, 1H), 3.74 - 3.63 (m , 3H), 3.16-3.48 (m, 1H), 2.41 (s, 6H), 2.30-2.19 (m, 2H).

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17 ZI '----( '----' m T.I. 410 411 δ 8.44 (s, 1H), 8.41 (s, 1H), 8.33 (s, 1H), 7.95 - 7.89 (m, 1H), 7.36 (dd, 7= 8 ,0, 2.4 Hz, 1H), 7.30 - 7.24 (m, 1H), 7.16-7.11 (m, 2H), 5.20 (s, 2H), 3.89 - 3.85 (m, 1H), 3.69 - 3.47 (m, 4H), 2.48 (s, 3H), 2.30-2.16 (m, 1H), 2.13 -2, 10 (m, 1H). 18 Y ^r °h ΠΊ 410 411 5 8.54 (s, 1H), 8.51 (s, 1H), 8.35 (s, 1H), 7.98 - 7.91 (m, 1H), 7.39 (dd, 7= 8 ,0, 2.4 Hz, 1H), 7.32 - 7.26 (m, 1H), 7.18 -7.13 (m, 2H), 5.22 (s, 2H), 3.89 ( t, 7= 4.0 Hz, 1H), 3.67 - 3.64 (m, 2H), 3.59 - 3.49 (m, 2H), 2.18 (s, 3H), 2.33 - 2.29 (m, 1H), 2.17-2.14 (m, 1H). 19 N-Г о Η Г ² ^ ⁿ y ⁿ yS 0 φ F 378 379 5 8.81 (t, 7= 3.6 Hz, 2H), 8.31 (s, 1H), 7.99 -7.95 (m, 2H), 7.53 (d, 7= 8.4 Hz, 1H), 7.41 (t, 7= 4.8 Hz, 1H), 7.24-7.15 (m, 3H), 5.13 (s, 2H), 3.90 - 3.87 (m, 1H), 3.78 - 3.72 (m, 2H), 3.65 - 3.60 (m, 1H), 3.56 3.50 (m, 1H), 2.38-2, 32 (m, 1H), 2.28 - 2.22 (m, 1H).

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19E1 YA N—< R or S H O F F 378 379 δ 8.81 (d, 7= 4.8 Hz, 2H), 8.32(s, 1H), 7.99 - 7.96 (m, 2H), 7.54 (d, 7= 8 Hz, 1H), 7.42 (t, 7= 4.8 Hz, 1H), 7.24 -7.19(m, 2H), 7.16 (d, 7 = 8 Hz, 1H), 5.14 ( s, 2H), 3.93 - 3.87 (m, 1H), 3.78 - 3.75 (m, 2H), 3.65 - 3.62 (m, 1H), 3.56 - 3, 50 (m, 1H), 2.38 -2.33 (m, 1H), 2.292.22 (m, 1H). 19E2 ^N (r or S ΛΊ h VN^N^A o NY Q F 378 379 δ 8.81 (d, 7= 4.8 Hz, 2H), 8.32(s, 1H), 7.99 - 7.96 (m, 2H), 7.54 (d, 7= 8 Hz, 1H), 7.42 (t, 7= 4.8 Hz, 1H), 7.24 -7.19(m, 2H), 7.16 (d, 7 = 8 Hz, 1H), 5.14 ( s, 2H), 3.93 -3.87 (m, 1H), 3.78 - 3.75 (m, 2H), 3.65 - 3.62 (m, 1H), 3.56 - 3.50 (m, 1H), 2 .38 -2.33 (m, 1H), 2.29- 2.22 (m, 1H). 20 F V.N N—Y O H ^VyS o N^J F F 396 397 δ 8.89 (s, 2H), 8.32 (s, 1H), 7.99 - 7.95 (m, 2H), 7.54 (d, 7 = 8 Hz, 1H), 7.24- 7.15 (m, 3H), 5.12 (s, 2H), 3.94 (t, 7 = 6 Hz, 1H), 3.80 - 3.72 (m, 2H), 3.63 - 3.60 (m, 1H), 3.56 - 3.50 (m, 1H), 2.37 - 2 .32 (m, 1H), 2.26-2.21 (m, 1H).

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20E1 E { ?N ^N \R or S О H Y ΥΊ о 9 F 396 397 δ 8.89 (s, 2H), 8.32 - 8.28 (m, 1H), 7.97 (dd, 7=8.8, 5.6 Hz, 2H), 7.54 (d, 7 =8.0 Hz, 1H), 7.22 (t, 7= 8.8 Hz, 2H), 7.16 (d, 7= 8.0 Hz, 1H), 5.13 (s, 2H), 3.93 - 3.89 (m, 1H), 3.80 -3.71(m, 2H), 3.65 - 3.59 (m, 1H), 3.56 - 3.52 (m, 1H ), 2.38 -2.34 (m, 1H), 2.272.21 (m, 1H). 20E2 E , N ^N -6 R or S /Y h AYY Ί o ^N A 9 F 396 397 δ 8.89 (s, 2H), 8.32 - 8.28 (m, 1H), 7.97 (dd, 7 = 8.8, 5.6 Hz, 2H), 7.54 (d,7= 8.0 Hz, 1H), 7.22 (t, 7= 8.8 Hz, 2H), 7.16 (d, 7= 8.0 Hz, 1H), 5.13 (s, 2H), 3.93 - 3.89 (m, 1H), 3.80-3.71 (m, 2H), 3.653.59 (m, 1H), 3.56 - 3.52 (m, 1H), 2.38 -2.34 (m, 1H), 2.27 -2.21 (m, 1H). 21 qXY °Y ZI OCY m 396 397 δ 8.89 (s, 2H), 8.34 (s, 1H), 7.93 - 7.88 (m, 1H), 7.41 (dd, 7=8, 2.4 Hz, 1H), 7.37-7.31 (m, 1H), 7.27 - 7.22 (m, 2H), 7.16 (d, 7= 8 Hz, 1H), 5.20 (d, 7 = 3, 6 Hz, 2H), 3.90 (1.7=7.2 Hz, 1H), 3.803.70 (m, 2H), 3.65 - 3.59 (m, 1H), 3.56 - 3, 52 (m, 1H), 2.38 - 2.32 (m, 1H), 2.25 - 2.20 (m, 1H).

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22 n-n Ca° L Η o nA S' F 412 413 δ 8.92 - 8.91 (m, 1H), 8.40 (s, 1H), 7.96 - 7.90 (m, 1H), 7.67 - 7.63 (m, 1H), 7 .39 - 7.37 (m, 1H), 7.32 - 7.25 (m, 2H), 7.17-7.13 (m, 2H), 5.77 (s, 1H), 5.22 (s, 2H), 3.83 (q,7= 4.0 Hz, 1H), 3.70 (q, 7=5.6 Hz, 2H), 3.59 - 3.55 (m, 1H) , 2.33 - 2.23 (m, 1H), 2.23 2.22 (m, 1H). 23 cu ^N H 7* A ⁿ yS 0 nA S' F 412 413 δ 8.64 (d, 7= 4.8 Hz, 2H), 8.39 (s, 1H), 7.98 - 7.91 (m, 1H), 7.39 -7.36 (m, 1H ), 7.31 -7.25 (m, 1H), 7.19-7.13 (m, 3H), 5.55 (s, 1H), 5.21 (s, 2H), 3.77 ( q, 7 = 3.6 Hz, 1H), 3.66 (t, 7 = 7.2 Hz, 2H), 3.58 - 3.54 (m, 1H), 2.29 2.53 (m, 1H), 2.20-2.17 (m, 1H). 24 n ZT AYA Π 410 411 6 8.74 (d, 7 = 5.2 Hz, 2H), 8.22 (s, 1H), 7.95 - 7.88 (m, 1H), 7.35 (t, 7 = 4.8 Hz, 2H), 7.29 -7.23 (m, 1H), 7.167.12 (m, 2H), 5.18 (s, 2H), 3.60 - 3.53 (m, 2H), 3 .39 3.35 (m, 1H), 3.12-3.07 (m, 1H), 3.04 - 2.94 (m, 2H), 2.76 -2.48 (m, 1H), 2.01 - 1.97 (m, 1H), 1.68 - 1.64 (m, 1H).

- 39 043804

25E1 V- /LR or S AR h ^VyS ο V F 410 411 5 8.76 (d, 7=4.8 Hz, 2H), 8.24 (s, 1H), 7.97 - 7.90 (m, 1H), 7.38 -7.36 (m, 2H ), 7.31 -7.25 (m, 1H), 7.18-7.13 (m, 2H), 5.20 (s, 2H), 3.63 3.53 (m, 2H), 3 .45-3.41 (m, 1H), 3.12 (t, 7=8.0 Hz, 1H), 3.04 - 2.95 (m, 2H), 2.78 2.75 (m, 1H), 2.05 - 2.02 (m, 1H), 1.70 - 1.66 (m, 1H). 25E2 ΖΛ / \ R or S ^Ν Λη _Η ΝΗ ₂ ο Ν^Ι V F 410 411 5 8.76 (d, 7= 4.8 Hz, 2H), 8.24 (s, 1H), 7.97 - 7.90 (m, 1H), 7.38 -7.36 (m, 2H ), 7.31 -7.25 (m, 1H), 7.18-7.13 (m, 2H), 5.20 (s, 2H), 3.63 3.53 (m, 2H), 3 .45-3.41 (m, 1H), 3.12 (t, 7= 8.0 Hz, 1H), 3.04 - 2.95 (m, 2H), 2.78 2.75 (m, 1H), 2.05 - 2.02 (m, 1H), 1.70 - 1.66 (m, 1H). 26 Μ ΖΙ Ύττ? -Π 410 411 5 9.11 (^7= 3.2 Hz, 1H), 8.27 (s, 1H), 7.97 - 7.90 (m,lH), 7.90 - 7.62 (m,2H) , 7.37 (dd, 7 =8.0, 2.0 Hz, 1H), 7.32 - 7.26 (m,lH), 7.18-7.13 (m,2H), 5.20 (s, 2H), 3.58 - 3.54 (m, 2H), 3.39 - 3.36 (m,lH), 3.16 - 3.04 (m,3H), 2.69 - 2 .67 (m, 1H), 2.00-1.98 (m,lH), 1.71 - 1.65 (m,lH).

- 40 043804

27 ^V <' ⁿ G ^! o U F 424 425 δ 8.58 (d, 7 = 5.2 Hz, 1H), 8.24 (s, 1H), 7.97 - 7.91 (m, 1H), 7.38 - 7.36 (m,lH ), 7.32 - 7.23 (m,2H), 7.78-7.13 (m, 2H), 5.01 (s, 2H), 3.61 - 3.53 (m,2H), 3.41 - 3.30 (m,lH), 3.12-3.08 (m,lH), 3.00-2.90 (m,2H), 2.77-2.51 (m,lH ), 2.44 (s, 3H), 2.12 - 1.96 (m, 1H), 1.73 - 1.59 (m,lH). 28 ή m ζζ “Π 424 425 δ 8.26 (s, 1H), 7.97 -7.90 (m, 1H), 7.54 -7.49 (m, 2H), 7.37 (dd, 7 = 8.0, 2, 4 Hz, 1H), 7.31-7.25 (m,lH), 7.18-7.13 (m,2H), 5.20 (s, 2H), 3.56 3.54 (m, 2H), 3.40 - 3.36 (m, 1H), 3.31 -3.10 (m,lH), 3.04 - 2.95 (m, 2H), 2.68 -2.63 ( m, 1H), 2.58 (s, 3H), 1.99 - 1.97 (m,lH), 1.71 -1.64 (m,lH). 29 M ZI —O-0-ι '-----< '-----' hj Π 414 415 δ 8.94 (d, 7= 5.2 Hz, 2H), 8.50 (s, 1H), 7.94 (t, 7= 7.2 Hz, 1H), 7.60 (t, 7= 4.4 Hz, 1H), 7.40 (q, 7=2.4 Hz, 1H), 7.30 (t, 7= 2.4 Hz, 1H), 7.16 (t, 7= 8, 0 Hz, 2H), 5.24 (s, 2H), 4.13 -4.02 (m, 2H), 3.87 (t, 7= 11.6 Hz, 1H), 3.66 (t, 7 = 4.8 Hz, 1H), 2.60 - 2.55 (m, 2H).

- 41 043804

29E1 Y^N O N G ^VyS about 7 F 414 415 δ 8.94 (d, J= 4.8 Hz, 2H), 8.51 (s, 1H), 7.98 -7.92 (m, 1H), 7.60 (t, J= 4.4 Hz, 1H), 7.40 (q, 7=2.4 Hz, 1H), 7.32 - 7.26 (m, 1H), 7.16 (t, 7 = 8.0 Hz, 2H), 5.25 (s, 2H), 4.13-4.02 (m, 2H), 3, 87 (t, 7 = 9.2 Hz, 1H), 3.67 (q.7 = 10.0 Hz, 1H), 2.71 -2.66(m, 2H). 29E2 Y'N ζ IF _c m-^xJ R or S o _H ύ ^VyS o V F 414 415 6 8.94 (d, 7 = 5.2 Hz, 2H), 8.51 (s, 1H), 7.98 -7.92 (m, 1H), 7.60 (t, 7 = 4.8 Hz, 1H), 7.40 (q, 7=2.4 Hz, 1H), 7.32 - 7.26 (m, 1H), 7.16 (t, 7 = 6.0 Hz, 2H), 5.25 (s, 2H), 4.13-4.05 (m, 2H), 3.88 (t, 7 = 8.8 Hz, 1H), 3.67 (q, 7 = 7.2 Hz , 1H), 2.74 - 2.57 (m, 2H). Y ZT ΎΥ ΊΊ 415 416 δ 8.38 (s, 1H), 7.97 - 7.91 (m, 1H), 7.41 - 7.37 (m, 2H), 7.32 -7.26 (m, 1H), 7 ,18-7.13 (m, 2H), 5.21 (s, 2H), 3.88 3.79 (m, 2H), 3.64 - 3.60 (m, 2H), 3.57 - 3.49 (m, 1H), 2.42 (s, 3H), 2.38-2.32 (m, 1H), 2.16-2.11 (m, 1H).

- 42 043804

31 ' O n o F 409 410 δ 8.46 (s, 1H), 8.32 (s, 1H), 8.41 (d, 7=3.6 Hz, 1H), 8.26 (s, 1H), 7.94 - 7, 88 (m, 1H), 7.66 (d, J= 7.6 Hz, 1H), 7.36 -7.23 (m, 3H), 7.15-7.12 (m, 2H), 5 .17 (s, 2H), 3.53 3.47 (m, 2H), 3.38 -3.30 (m, 2H), 3.08 - 3.04 (m, 1H), 2.72 - 2.69 (m, 2H), 1.93 - 1.92 (m, 1H), 1.63 - 1.58 (m, 1H). 32 \il ^z “P 416 417 δ 8.43 (s, 1H), 7.97 - 7.91 (m, 1H), 7.40 - 7.38 (m, 1H), 7.32 -7.26 (m, 1H), 7 ,18-7.14 (m, 2H), 5.23 (s, 2H), 4.01 3.89 (m, 2H), 3.67 - 3.58 (m, 2H), 3.56 - 3.52 (m, 1H), 2.71 (s, 3H), 2.45-2.41 (m, 1H), 2.19-2.14 (m, 1H). 33 LL W J-----1 J-----1 IZ >O ,-z ,z/ i ^X^Z 410 411 δ 8.60 (s, 2H), 8.01 - 7.95 (m, 1H), 7.46 (dd,7=8.4, 6.4 Hz, 1H), 7.20-7.16 (m, 2H), 7.07 - 6.99 (m, 2H), 4.73 (s, 2H), 3.98 - 3.93 (m, 1H), 3.75 3.69 (m, 1H), 3.59 -3.35 (m, 1H), 3.47 - 3.42 (m, 2H), 2.62 (s, 3H), 2.41 -2.38(m, 1H) , 2.10-2.09 (m, 1H).

- 43 043804

34 O.J.-----. \-----. IZ th 398 399 δ 8.00 - 7.93 (m, 1H), 7.50 (s, 1H), 7.44 (d, 7=7.2 Hz, 1H), 7.16 (d, 7=8.0 Hz, 1H), 7.096.98 (m, 4H), 6.95 (s,lH), 4.70 (s, 2H), 4.03 (d, 7=8.4 Hz, 2H), 3, 57 - 3.50 (m, 2H), 3.46-3.40 (m, 1H), 3.203.16 (m, 1H), 2.71 (s, 1H), 2.06 - 1.99 ( m, 1H), 1.73 1.68 (m, 1H). 35 Pz z ZI Π 398 399 δ 8.00 - 7.94 (m, 1H), 7.55 7.43 (m, 3H), 7.16 (d, 7= 8.0 Hz, 1H), 7.07 - 6.98 ( m, 3H), 6.25 (s, 1H), 4.71 (s, 2H), 4.19 -4.17(m, 2H), 3.58 - 3.42 (m, 3H), 3 .25 - 3.20 (m, 1H), 2.83 -2.78 (m, 1H), 2.04 - 1.93 (m, 1H), 1.76-1.71 (m, 1H) . 36 - ^Z \A Z—' °4 ZI ΆΗ7Η “P 412 413 δ 8.25 (s, 1H), 7.92 - 7.91 (m, 1H), 7.36 (dd, 7= 6.0, 2.4 Hz, 1H), 7.31-7, 25 (m, 1H), 7.17 -7.13 (m, 2H), 7.01 (s, 1H), 6.75 (s, 1H), 5.21 (s, 2H), 3.63 (q, 7=6.8 Hz, 1H), 3.61 (s, 3H), 3.57-3.51 (m, 1H), 3.42 -3.38(m, 1H), 3, 11 (t.7=8.4 Hz, 1H), 2.77 - 2.72 (m, 2H), 2.70 - 2.62 (m, 1H), 2.06 2.04 (m, 1H ), 1.70 - 1.65 (m, 1H).

- 44 043804

37 F ^F A AY AHA vvyS 0 nJ Φ F 446 447 δ 8.93 (s, 1H), 8.83 (s, 1H), 8.37 (s, 1H), 7.98 - 7.91 (m, 1H), 7.38 (dd J =8, 4, 2.4 Hz, 1H), 7.31-6.99 (m, 4H), 5.23 (s, 2H), 3.96 - 3.92 (m, 1H), 3.83 - 3 .79 (m, 1H), 3.69 3.62 (m, 2H), 3.57-3.51 (m, 1H), 2.42-2.35 (m, 1H), 2.24 - 2.15 (m, 1H). 39 Ajj ^N ( R or S O HY vn^n^ TY η o ^N J As F 384 385 5 8.80 (d, 7= 4.8 Hz, 2H), 8.29 (s, 1H), 7.48 (d, 7= 8.4 Hz, 1H), 7.41 (t, 7= 4.8 Hz, 1H), 7.16 (t, 7=4.0 Hz, 1H), 7.11 (d, 7=8.0 Hz, 1H), 6.66 (q, 7= 2, 0 Hz, 1H), 5.11 (s, 2H), 3.91 -3.85 (m, 1H), 3.77 - 3.71 (m, 2H), 3.63 - 3.57 (m , 1H), 3.54 - 3.48 (m, 1H), 2.37-2.31 (m, 1H), 2.282.21 (m, 1H). 40 ^N AR or S Al H Ύ 0 N^J 4 F 384 385 5 8.80 (d, 7= 4.8 Hz, 2H), 8.29 (s, 1H), 7.48 (d, 7= 8.4 Hz, 1H), 7.41 (t, 7= 4.8 Hz, 1H), 7.16 (t, 7= 4.0 Hz, 1H), 7.11 (d, 7= 8.0 Hz, 1H), 6.66 (q, 7= 2, 0 Hz, 1H), 5.11 (s, 2H), 3.91 - 3.85 (m, 1H), 3.77 - 3.71 (m, 2H), 3.63 - 3.57 (m, 1H), 3.54 -3.48(m, 1H), 2.37-2.31 (m, 1H), 2.28-2.21 ( m, 1H).

- 45 043804

41 Y and 0 N^l 6 ^S 366 367 5 8.80 (q, J= 3.2 Hz, 2H), 8.29 (s, 1H), 7.49 - 7.46 (m, 2H), 7.42 - 7.39 (m, 2H ), 7.12 (d, 7=8.0 Hz, 1H), 7.06 (t, 7 = 3.6 Hz, 1H), 5.10 (s, 2H), 3.87 (d.7 = 10.4 Hz, 1H), 3.75 (d, 7 = 6.4 Hz, 2H), 3.61 (t, 7 = 3.2 Hz, 1H), 3.52 (t, 7 = 9 .6 Hz, 1H), 2.36 -2.22 (m, 2H). 42 z—'°=< zz сЛ ^И * 400 401 5 8.80 (d, 7= 4.8 Hz, 2H), 8.28 (s, 1H), 7.48 (d,7= 8.4 Hz, 1H), 7.41 (t, 7= 4.8 Hz, 1H), 7.33 (d, 7= 4.0 Hz, 1H), 7.12 (t, 7= 4.4 Hz, 1H), 7.05 (d, 7= 4, 0 Hz, 1H), 5.19 (s, 2H), 3.67 (d,7= 11.2 Hz, 1H), 3.74 (d, 7=5.2 Hz, 2H), 3.59 (d, 7 = 8.4 Hz, 1H), 3.52 (t, 7 = 6.8 Hz, 1H), 2.37 - 2.24 (m, 2H). 43 ь ^N ZR or S o NyJ F 402 403 5 8.89 (s, 2H), 8.29 (s, 1H), 7.47 (d, 7=4.0 Hz, 1H), 7.177.10 (m, 2H), 6.66 (q, 7= 2.0 Hz, 1H), 5.12 (s,2H), 3.903.86 (m, 1H), 3.79 - 3.69 (m, 2H), 3.61 -3.47 (m , 2H), 2.37 -2.33 (m, 1H), 2.24-2.19 (m, 1H).

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44 F ί/N N—5 R or S Q F 402 403 δ 8.89 (s, 2H), 8.29 (s, 1H), 7.47 (d, 7=4.0 Hz, 1H), 7.177.10 (m, 2H), 6.66 (q, 7=2.0 Hz, 1H), 5.12 (s, 2H), 3.903.86 (m, 1H), 3.79 - 3.69 (m, 2H), 3.61 -3.47 (m , 2H), 2.37 -2.33 (m, 1H), 2.24-2.19 (m, 1H). 45 )n _F D H o nY 4 402 403 5 8.95 (d, 7=5.2 Hz, 2H), 8.46 (s, 1H), 7.60 (t, 7= 8.4 Hz, 1H), 7.49 (d, 7= 8.4 Hz, 1H), 7.17(t, 7= 4.0 Hz, 1H), 7.12 (d, 7= 8.0 Hz, 1H), 6.68-6.66 (m, 1H), 5.15 (s, 2H), 4.124.04 (m, 2H), 3.86 (t, 7= 10.0 Hz, 1H), 3.66 (t, 7= 8.0 Hz, 1H), 2.68 - 2.54 (m, 2H). 46 0 ^H T YY η ⁰ Φ F 381 382 5 8.35 (s, 1H), 7.97 (q,7=6.4 Hz, 2H), 7.54 (d, 7=8.4 Hz, 1H), 7.24-7.15 ( m, 3H), 6.76 (s, 1H), 5.13 (s, 2H), 3.81 (t, 7 = 6.4 Hz, 1H), 3.68 - 3.59 (m, 2H ), 3.57 - 3.60 (m, 2H), 2.33 - 2.28 (m, 1H), 2.25 (s, 3H), 2.20-2.15 (m, 1H). 47 Y Η Г ² ^YyS о 9' F 413 414 CD3OD as solvent δ 7.92 - 7.88 (m, 2H), 7.48 (d, 7=8 Hz, 1H), 7.26 (d, 7=8.4 Hz, 1H), 7.14 -7.10 (m, 2H), 6.69 (s, 1H) 3.78 - 3.71 (m, 1H), 3.67 - 3.61 (m, 1H), 3.54 -3, 47(m, 1H), 3.26-3.21 (m, 1H), 2.89 (d, 7=7.2 Hz, 2H), 2.78 - 2.73 (m, 1H), 2 ,thirty

- 47 043804

(d, 7= 0.8 Hz, ZN), 2.172.14 (m, 1H), 1.84 - 1.77 (m, 1H). 48 Ν-Ϋ X n 0 N^J σ' 378 379 5 8.80 (d, 7=5.2 Hz, 2H), 8.33 (s, 1H), 7.93 - 7.89 (m, 1H), 7.43 - 7.36 (m, 2H ), 7.36 -7.31 (m, 1H), 7.27-7.22 (m, 2H), 7.16 (d, 7= 8.4 Hz, 1H), 5.22 (s, 2H), 3.90 - 3.87 (m, 1H), 3.78 - 3.75 (m, 2H), 3.63 - 3.60 (m, 1H), 3.56 3.50 (m , 1H), 2.37 -2.33 (m, 1H), 2.27 - 2.22 (m, 1H). 49 F Ν— Λη _Η νη ₂ Ο _υ ^ν Α 0 0 ^s 384 385 5 8.78 (s, 2H), 8.18 (s, 1H), 7.38 -7.35 (m, 2H), 7.30 (dd, 7=5.2, 0.8 Hz, 1H ), 7.00 (d, 7 = 8.4 Hz, 1H), 6.94 (dd,7 = 5.2, 3.6 Hz, 1H), 4.99 (s, 2H), 3.80 - 3.75 (m, 1H), 3.68 3.58 (m, 2H), 3.48 - 3.46 (m, 1H), 3.42 - 3.37 (m, 1H), 2, 28 -2.19(m, 1H), 2.16-2.01(m, 1H),

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8.63 (d, 7=5.2 Hz, 1H), 8.30 (s, 1H), 7.99 - 7.95 (m, 2H), 7.53 (d, 7=8.0 Hz , 1H), 7.27 (d, 7=5.2 Hz, 1H), 7.24-7.19 (m, 2H), 7.15 (d, 7= 8.0 Hz, 1H), 5 .12 (s, 2H), 3.90 - 3.85 (m, 1H), 3.76 - 3.61 (m, 3H), 3.54 - 3.52 (m, 1H), 2.47 (s, 3H), 2.33 - 2.26 (m, 2H).

8.66 (d, 7= 5.2 Hz, 1H), 8.28 (s, 1H), 7.97 - 7.93 (m, 2H), 7.51 (d, 7= 8.4 Hz , 1H), 7.25 (d, 7=5.2 Hz, 1H), 7.21 7.17 (m, 2H), 7.14 (d, 7= 8.0 Hz, 1H), 5, 10 (s, 2H), 3.883.84 (m, 1H), 3.74 - 3.59 (m, 3H), 3.52 - 3.46 (m, 1H), 2.45 (s, 3H) , 2.32-2.19 (m, 2H).

9.02 (s, 2H), 8.47 (s, 1H), 7.97 - 7.94 (m, 2H), 7.53 (d, 7 = 8.4 Hz, 1H), 7.24 -7.14 (m, 3H), 5.10 (s,2H), 4.12-3.98 (m, 2H), 3.85 (1.7 = 8.4 Hz, 1H), 3, 69 - 3.62 (m, 1H), 2.68 - 2.55 (m, 2H).

δ 9.02 (s, 2H), 8.47 (s, 1H), 7.97 - 7.94 (m, 2H), 7.53 (d, J = 8.4 Hz, 1H), 7, 22-7.14 (m, 3H), 5.10 (s, 2H), 4.12-4.01 (m, 2H), 3.85 (t, 7=8.4 Hz, 1H), 3 .69 - 3.62 (m, 1H), 2.66 - 2.55 (m, 2H).

8.95 (d, 7=5.2 Hz, 2H), 8.49 (s, 1H), 7.98 (t, 7= 8.4 Hz, 2H), 7.62 - 7.55 (m , 2H), 7.24-7.16 (m, 3H), 5.13 (s, 2H), 4.14-4.04 (m, 2H), 3.90 -3.86(m, 1H ), 3.68(1.7=5.2 Hz, 1H), 2.67 - 2.57 (m, 2H).

8.95 (d, 7=5.2 Hz, 2H), 8.49 (s, 1H), 7.98 (t,7= 8.4 Hz, 2H), 7.62-7.55 (m , 2H), 7.24-7.16 (m, 3H), 5.13 (s, 2H), 4.14-4.04 (m, 2H), 3.90 -3.86 (m, 1H ), 3.68 (t, 7=5.2 Hz, 1H), 2.67 - 2.57 (m, 2H).

Enzymatic analysis of HDAC2 and HDAC1 (IC50 data for HDAC2 and HDAC1).

An assay protocol for measuring the deacetylation of a substrate peptide using

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HDAC2 or HDAC1.

The HDAC protein composition and corresponding substrate peptides are listed below.

Analysis name Expressive construct Regulatory subunit Peptide substrate HDAC1 Full-length human HDAC 1 with a C-terminal His tag and a C-terminal FLAG tag expressed in a baculovirus expression system. Absent FAMTSRHK(Ac)KLNH2 HDAC2 Full-length human HDAC2 with a C-terminal FLAG tag expressed in a baculovirus expression system. Absent FAMTSRHK(Ac)KLNH2

Preparation of analysis.

HDAC reactions were collected in 384-well plates (Greiner) in a total volume of 20 μl as follows.

HDAC proteins are pre-diluted in assay buffer containing 100 mM HEPES, pH 7.5, 0.1% BSA, 0.01% Triton X-100, 25 mM KCl and dispensed into a 384-well plate (10 µl per well). .

Test compounds are serially pre-diluted in DMSO and added to protein samples by acoustic pipetting (Labcyte Echo). The DMSO concentration in all samples was adjusted to 1%.

Control samples (0% inhibition in the absence of inhibitor, DMSO only) and 100% inhibition (in the absence of enzyme) were collected in quadruplicate and used to calculate %inhibition in the presence of compounds.

At this stage, compounds can be pre-incubated with enzyme if necessary.

Reactions are initiated by adding 10 μl of FAM-labeled substrate peptide previously diluted in the same assay buffer. The final concentration of the substrate peptide is 1 μM (HDAC 1-2). The reactions are allowed to proceed at room temperature. After incubation, reactions were quenched by adding 50 μl of termination buffer (100 mM HEPES, pH 7.5, 0.01% Triton X-100, 0.1% SDS). The end plates are analyzed on a microfluidic electrophoresis instrument (Caliper LabChip® 3000, Caliper Life Sciences/Perkin Elmer), which allows electrophoretic separation of the deacetylated product from the acetylated substrate. The measured parameter is the change in the relative intensity of the substrate peptide and the product. The activity in each test sample is defined as the product-to-sum ratio (PSR): P/(S+P), where P is the product peak height and S is the substrate peak height. Percent inhibition (P _Inhibition ) is determined using the following equation:

^{Ring (PSR} 0%ing ^-PSR compound ^)/(PSR 0%ing ^-PSR 100%ing ^{)x 100} where the PSR _{of the compound} is the product/total ratio in the presence of the compound;

PSR ₀ % _ing is the product/total ratio in the absence of compound; And

PSR ₁₀₀ % _ing is the product/total ratio in the absence of enzyme.

To determine the IC50 of compounds (50% inhibition), %ing data ( _Ring relative to compound concentration) are fit to a 4-parametric sigmoidal dose-response model using XLfit software (IDBS).

The results of this analysis for certain compounds are presented in table. 2 below. Table A indicates an IC50 value of less than 0.5 µM; B - for IC50 value from 0.5 µM to 1.0 µM; C - for an IC50 value greater than 1.0 µM and less than or equal to 2.0 µM; and D indicates an IC50 value greater than 2.0 μM. NT=not tested.

table 2

Compound No. IC50 for HDAC2, (µM) IC50 for HDAC1, (µM) Compound No. IC50 for HDAC2, (µM) IC50 for HDAC1, (µM)

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1 WITH V 2 D WITH 3 D D 4 WITH V 5 NT NT 6 NT NT 7 IN A 8E1 A A 8E2 WITH WITH 9 WITH V 10 V A eleven nt NT 12 nt NT 13 nt nt 14 V V 14E1 V A 14E2 D V 15 NT nt 16 nt nt 17 nt nt 18 V V 19 A A 19E1 A V 19E2 A A 20 V A 20E1 V V 20E2 A A 21 WITH V 22 nt NT 23 D D 24 WITH WITH 25E1 WITH WITH 25E2 D WITH

26 WITH With 27 NT nt 28 nt nt 29 V A 29E1 A A 29E2 WITH WITH thirty nt nt 31 nt nt 32 nt nt 33 nt nt 34 nt nt 35 nt nt 36 V A 37 nt nt 39 With With 40 A A 41 V A 42 nt NT 43 D With 44 V A 45 With V 46 With V 47 nt nt 48 With V 49 With A 50 With 1 51 V A 52 With V 53 V A 54 V V 55 A A

HDAC2 enzymatic inhibition assay in SH-SY5Y cell lysate with exogenous substrate

SH-SY5Y cells (Sigma) were cultured in modified Eagle's essential medium supplemented with 10% fetal bovine serum and pen/strap. 24 hours before compound dosing, 20 μl of cells were seeded into white 384-well plates at a density of 1500 cells/well. Compounds were serially diluted in neat DMSO and then diluted 1:100 v/v. in FBS-free medium and mixed. The media was removed from the cells on the plates, diluted compounds in serum-free medium (1% v/v final DMSO) were added and incubated at 37°C for 5 hours. Then 10 μl of HDAC-Glo2 reagent with 0.1% Triton X was added -100, the plate was stirred and allowed to develop at room temperature for 100 m. The plates were then read using a Spectramax LMax luminometer using an integration time of 0.4 s. Dose-response curves were generated with normalized data, where CI-994 at 100 μM was defined as 100% inhibition and pure DMSO as 0% inhibition.

The results of this analysis for certain compounds are presented in table. 3 below. Table A indicates an IC50 value of less than 0.5 µM; B represents an IC50 value of 0.5 to 1.0 μM; C represents an IC50 value greater than 1.0 μM and less than or equal to 2.0 μM; and D indicates an IC50 value greater than 2.0 μM. NT=not tested.

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Compound No. IC50 vs. HDAC2, SHSY5Y cell lysate (µM) 1 D 2 D 3 D 4 D 5 D 6 D 7 IN 8E2 D 8E2 IN 9 WITH 10 WITH eleven D 12 D 13 D 14 WITH 14E1 IN 14E2 D 15 D 16 D 17 WITH 18 IN 19 A 19E1 WITH 19E2 IN 20 WITH 20E1 With 20E2 V 21 With 22 D 23 D 24 WITH 25E1 IN 25E2 IN

Table 3

Compound No. IC50 vs. HDAC2, SHSY5Y cell lysate (µM) 26 IN 27 WITH 28 With 29 V 29E1 V 29E2 With thirty D 31 D 32 WITH 33 D 34 D 35 D 36 IN 37 D 39 D 40 IN 41 IN 42 D 43 WITH 44 IN 45 IN 46 D 47 D 48 WITH 49 WITH 50 D 51 WITH 52 D 53 IN 54 WITH 55 IN

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Comparison of pyrrolidine rings substituted at position 3 with heteroaromatic rings or attached through a methylene group with heteroaromatic rings with pyrrolidines substituted with non-aromatic groups at position 3 for pyrrolidine ureas.

In table 4 below shows a direct comparison of the activity levels between certain compounds having a non-aromatic substitution at the 3-position of the pyrrolidinyl moiety and the compounds of the present invention having an aromatic substitution at the 3-position of the pyrrolidinyl moiety (ie R ¹ with or without an X spacer group). The data show that increased efficiency in the HDAC2 activity assay in SH-SY5Y cell lysate occurs when the non-cyclic, non-aromatic R ¹ substituents in reference compounds AC are replaced by aromatic pyrimidinyl in compounds 19 and 20. A similar trend is shown for the other compounds in Table 1. 4. Compound 20, with 5-F-pyrimidine directly linked to position 3, is more than 2 times more potent than reference compound A.

Table 4

No. Structure IC50 for HDAC2, SH-SY5Y cell lysate (µM) Comparison connection A /0 n ⁰ QF 2.64 Comparison connection B ΤΙ ZI 14 Comparison connection C o N 0 F F 6.36 19 \ T ^z z— ZI 0.421

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0 F Ъ ⁿ G O f F 1.13 18 0% ZI '----( ⁴ ----' Yu n 0.550 29 IZ >=O ^z 0.768 Comparison connection E 24 26 O.J.-----. U----- _v CJ y-----. \-----OJ J-----. \ ь оIZ IZ IZ )=O )=O )=O A :i ^} >100 1.42 0.535

The contents of all references (including literary references, issued patents, published patent applications and pending patent applications) mentioned in this application are expressly incorporated herein in their entirety by reference. Unless otherwise specified, all technical and scientific terms used in this application have the meanings commonly known to one skilled in the art.

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Claims (20)

CLAIM 1. Compound of formula III or IIIa R ⁴ (III); or ^R 3 (Sha); or a pharmaceutically acceptable salt thereof, wherein X is (CR ^a R ^b ) _t , where t is 0 or 1 or O; q represents 0 or 1; R ¹ is phenyl, pyridinyl, pyrazinyl, pyridazinyl, pyrimidinyl, thiazolyl, thiadiazolyl, imidazolyl, pyrazolyl or oxazolyl, each of which is optionally substituted with 1 to 3 groups selected from R ^c ; R ² represents halogen; R ³ represents hydrogen or halogen; R ⁴ represents halogen; R ^a and R ^b each represent hydrogen; R ^c represents halogen, halogen(C ₁ -C ₄ )alkyl, (C ₁ -C ₄ )alkyl or (C ₁ -C ₄ )alkylO(C ₁ -C ₄ )alkyl. 2. A compound according to claim 1, characterized in that the compound is a compound of formula IV or IVa or a pharmaceutically acceptable salt thereof. 3. A compound according to claim 1 or 2, characterized in that R ³ is halogen. 4. A compound according to any one of claims 1 to 3, characterized in that R ³ represents fluorine. 5. A compound according to claim 1 or 2, characterized in that R ³ represents hydrogen. 6. A compound according to any one of claims 1 to 5, characterized in that R ⁴ represents fluorine. 7. A compound according to any one of claims 1 to 6, characterized in that X is (CR ^a R ^b ). 8. A compound according to any one of claims 1 to 7, characterized in that the compound is a compound of formula V or Va or a pharmaceutically acceptable salt thereof. 9. A compound according to any one of claims 1 to 7, characterized in that the compound is a compound of formula VI or VIa - 55 043804 or a pharmaceutically acceptable salt thereof. 10. A compound according to any one of claims 1 to 7, characterized in that the compound is a compound of formula VII or VIIa or a pharmaceutically acceptable salt thereof. 11. A compound according to any one of claims 1 to 10, characterized in that R ^c represents fluorine, methyl, CHF2 or CH2OCH3. 12. The connection according to claim 1, characterized in that the connection is selected from - 56 043804 - 57 043804 or pharmaceutically acceptable salts thereof. 13. The connection according to claim 1, characterized in that the connection is selected from - 58 043804 - 59 043804 - 60 043804 or pharmaceutically acceptable salts thereof. 14. A pharmaceutical composition containing a compound according to any one of claims 1 to 13 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. 15. Use of a compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof, or a composition according to claim 14 for inhibiting histone deacetylase (HDAC) activity in a subject. 16. Use of a compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof, or a composition according to claim 14, for the treatment of a condition in a subject selected from a neurological disorder, a disorder or impairment of cognitive function, a fungal infection, an inflammatory disease, a mental disorder and neoplastic diseases. - 61 043804 17. Application according to claim 16, characterized in that the state is a) disorder or impairment of cognitive function associated with Alzheimer's disease, posterior cortical atrophy, normal pressure hydrocephalus, Huntington's disease, seizure-induced memory loss, schizophrenia, Rubinstein-Taybi syndrome, Rett syndrome, depression, fragile X syndrome, dementia with body cells Lewy, stroke, vascular dementia, vascular cognitive impairment (VCI), Binswanger's disease, frontotemporal lobar degeneration (FTLD), ADHD, dyslexia, major depressive disorder, bipolar disorder and social, cognitive and learning disorders associated with autism, traumatic brain injury (TBI), chronic traumatic encephalopathy (CTE), multiple sclerosis (MS), attention deficit disorder, anxiety disorder, conditioned fear, panic disorder, obsessive-compulsive disorder, post-traumatic stress disorder (PTSD), phobia, social anxiety disorder, recovery from drug addiction, age-related memory impairment (AAMI), age-related cognitive decline (ARCD), ataxia, Parkinson's disease or Parkinson's disease dementia; or b) a hematological disease selected from acute myeloid leukemia, acute promyelocytic leukemia, acute lymphoblastic leukemia, chronic myelogenous leukemia, myelodysplastic syndromes and sickle cell anemia; or c) neoplastic disease; or d) disorder of learning extinction, selected from fear cessation and post-traumatic stress disorder; or e) hearing loss or hearing impairment; or f) fibrotic diseases such as pulmonary fibrosis, renal fibrosis, cardiac fibrosis and scleroderma; or g) bone pain in patients with cancer; or h) neuropathic pain. 18. The use of claim 17, wherein the condition is Alzheimer's disease, Huntington's disease, frontotemporal dementia, Friedreich's ataxia, post-traumatic stress disorder (PTSD), Parkinson's disease, or recovery from drug addiction. 19. The use of claim 16, wherein the condition is selected from Alzheimer's disease, Huntington's disease, frontotemporal lobar degeneration, Friedreich's ataxia, post-traumatic stress disorder, Parkinson's disease, Parkinson's disease dementia, drug addiction recovery, disorder or disorder cognitive function, neurological disorder with synaptic pathology, disorder of learning discrimination, mental disorders, cognitive impairment or disorders associated with Alzheimer's disease, dementia with Lewy bodies, schizophrenia, Rubinstein-Taybi syndrome, Rett syndrome, syndrome fragile X, multiple sclerosis, age-related memory impairment, age-related cognitive decline, and social, cognitive and learning disorders associated with autism. 20. The use of claim 16, wherein the condition is selected from a memory disorder or disorder, a fungal disease, an extinction learning disorder, and a hematological disease. Eurasian Patent Organization, EAPO Russia, 109012, Moscow, Maly Cherkassky lane, 2