RU2773516C2 - NEW IMIDAZO[4,5-c]QUINOLINE DERIVATIVES AS LRRK2 INHIBITORS - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: invention relates to compounds of formula I and their pharmaceutically acceptable salts having inhibitory activity against leucine-rich repeat kinase 2 (LRRK2). In formula I, R1 is selected from the group consisting of methyl, ethyl, cyclobutyl, cyclopentyl, etc., R2 is selected from the group consisting of 2,2-difluoropropyl, etc., R3 is selected from the group consisting of fluorine, chlorine, cyano, difluoromethyl and trifluoromethyl, wherein the compounds have one or more than one hydrogen atom which is replaced by a deuterium atom, provided that one or more hydrogen atoms are not replaced only by a deuterium atom in one or more of positions 2, 5, 7 or 8 of the quinoline ring. The invention also relates to specific compounds, in particular to [5-({8-chloro-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[4,5-c ]quinolin-2-yl}methyl)pyrazin-2-yl]methanol and 8-chloro-2-{[5-(2H3)methylpyrazin-2-yl]methyl}-1-[(2R,4R)-2- methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[4,5-c]quinoline, a pharmaceutical composition containing said compounds, and a method for treating a disease or disorder selected from the group consisting of Crohn's disease, Parkinson's disease, dementia with Lewy bodies, leprosy, Alzheimer's disease and tauopathy, with their use.

EFFECT: Crohn's disease, Parkinson's disease, dementia with Lewy bodies, leprosy, Alzheimer's disease and tauopathy treatment method improvement.

19 cl, 6 tbl, 96 ex

Description

FIELD OF THE INVENTION

The present invention relates to small molecule inhibitors of leucine repeat-rich kinase 2 (LRRK2). This invention also relates to methods for inhibiting LRRK2 in mammals, including humans, by administering small molecule inhibitors of LRRK2. The present invention also relates to the treatment of Parkinson's disease (PD) and other neurodegenerative and/or neurological disorders in mammals, including humans, with LRRK2 inhibitors. More specifically, this invention relates to novel imidazo[4,5-c]quinoline compounds useful in the treatment of neurodegenerative and/or neurological disorders such as PD, Alzheimer's disease (AD) and other LRRK2-associated disorders.

PRIOR ART

LRRK2 is a 286 kDa protein in the ROCO protein family with a multidomain complex structure. Protein motifs that have been identified for the LRRK2 protein include the armadil-like (ARM) domain, the ankyrin-like (ANK) domain, the leucine repeat-rich (LRR) domain, the Ras domain (the renin-angiotensin system) complex (ROC), the C-terminal ROC (COR ) domain, kinase domain and C-terminal domain of WD40. The ROC domain binds guanosine triphosphate (GTP), and the COR domain may be a regulator of the GTPase activity of the ROC domain. The kinase domain shares structural homology with MAPkinase kinase (MAPKKK) kinases and has been shown to phosphorylate a number of cellular proteins in vitro, however the endogenous substrate has yet to be determined. LRRK2 has been found in various regions of the brain, as well as in a number of peripheral tissues, including the heart, lung, spleen, and kidney.

Due to its multi-domain design, LRRK2 appears to have the ability to play complex roles in a variety of cellular processes, each associated with putative protein-protein interactions, guanosine triphosphatase (GTPase) activity, and kinase activity. For example, LRRK2 is associated with inhibition of nuclear factor activated T lymphocytes (NFAT) in the immune system and is associated with vesicle migration, presynaptic homeostasis, mammalian target of rapamycin (mTOR) signaling, MET receptor tyrosine kinase signaling in papillary renal carcinomas and thyroid carcinomas. gland, dynamic characteristics of the cytoskeleton, the mitogen-activated protein kinase (MAPK) signaling pathway, the tumor necrosis factor-α (TNF-α) pathway, the Wnt pathway, and autophagy. According to the latest genome-wide genetic association (GWA) studies, LRRK2 is implicated in the pathogenesis of various human diseases such as PD, inflammatory bowel disease (Crohn's disease), cancer, and leprosy (Lewis, P.A. and Manzoni, C. Science Signaling 2012, 5(207) , pe2).

Parkinson's disease (PD) is a relatively common age-related neurodegenerative disorder resulting from the progressive loss of dopamine-producing neurons, affecting up to 4% of the population over 80 years of age. PD is characterized by both motor symptoms such as resting tremor, rigidity, akinesia, and postural unsteadiness, as well as non-motor symptoms such as impaired cognition, sleep, and sense of smell. GWA studies have shown an association of LRRK2 with PD, and many patients with point mutations in LRRK2 exhibit symptoms that cannot be distinguished from those of patients with idiopathic PD. More than 20 LRRK2 mutations are associated with autosomal dominant parkinsonism, and the R1441C, R1441G, R1441H, Y1699C, G2019S, I2020T, and N1437H missense mutations are considered pathogenic. The LRRK2 R1441G mutation has been shown to increase the release of pro-inflammatory cytokines (higher levels of TNF-α, IL-1β, IL-12 and lower levels of IL-10) in microglial cells from transgenic mice, and thus may lead to direct toxicity in neurons (Gillardon, F. et al. Neuroscience 2012, 208, 41-48). In a mouse model of neuroinflammation, LRRK2 induction in microglia has been identified, and inhibition of LRRK2 kinase activity by small molecule inhibitors of LRRK2 (LRRK2-IN-1 or sunitinib) or knockout of the LRRK2 gene resulted in attenuated TNF-α secretion and induction of nitric oxide synthase (iNOS) ( Moehle, M. et al J. Neurosci 2012, 32(5), 1602-1611). The most common of the LRRK2 mutations, G2019S, is present in more than 85% of PD patients who have LRRK2 mutations. This mutation, which is present in the LRRK2 kinase domain, results in an increase in LRRK2 kinase activity. In the human brain, LRRK2 expression is highest in the same brain regions that are affected in PD patients, and LRRK2 is found in Lewy bodies, a hallmark of PD. Recent studies indicate that a potent selective brain-penetrating LRRK2 kinase inhibitor may be a therapeutic agent for the treatment of PD.

Dementia is the result of a wide variety of different pathological processes. The most common pathological processes causing dementia are AD, cerebral amyloid angiopathy (CM), and prion-mediated diseases (see, for example, Haan et al., Clin. Neurol. Neurosurg. 1990, 92(4): 305-310; Glenner et al., J. Neurol Sci. 1989, 94: 1-28). AD is a progressive neurodegenerative disorder characterized by memory impairment and cognitive dysfunction. AD affects about half of all people over the age of 85, the fastest growing part of the US population. In view of this, the number of patients with AD in the US is expected to increase from about 4 million to about 14 million by 2050. LRRK2 mutations are associated with AD-like pathology, suggesting that there may be overlap between neurodegenerative pathways in both AD and PD (Zimprach, A. et al. Neuron 2004, 44, 601-607). In addition, the LRRK2 R1628P variant (COR domain) is associated with an increased incidence of AD in a certain population, probably as a result of increased apoptosis and cell death (Zhao, Y. et al.; Neurobiology of Aging 2011, 32, 1990-1993).

An increase in the incidence of some non-skin cancers such as kidney, breast, lung, and prostate cancers, as well as acute myeloid leukemia (AML) has been reported in patients with Parkinson's disease with the LRRK2 G2019S mutation (Saunders-Pullman, R. et al. ; Movement Disorders, 2010, 25(15), 2536-2541). Since the G2019S mutation is associated with increased LRRK2 kinase activity, inhibition of this activity may be useful in the treatment of cancer such as kidney, breast, lung, prostate, and blood cancers.

Inflammatory bowel disease (IBD) or Crohn's disease (CD) is a complex disease and is believed to be the result of an inadequate immune response to the microbiota in the intestinal tract. GWA studies have recently identified LRRK2 as a major susceptibility gene for Crohn's disease, specifically the M2397T polymorphism in the WD40 domain (Liu, Z. et al. Nat. Immunol. 2011, 12, 1063-1070). In a recent study, LRRK2-deficient mice were found to be more susceptible to dextran sulfate-induced colitis compared to wild-type mice, indicating that LRRK2 may play a role in the pathogenesis of IBD (Liu, Z. and Lenardo, M. ; Cell Research 2012, 1-3).

Both non-selective and selective small molecule compounds with LRRK2 inhibitory activity have been described, such as staurosporine, sunitinib, LRRK2-IN-1, CZC-25146, TAE684 and described in WO 2011/141756, WO 2012/028629 and WO 2012/ 058193. It is desirable to provide compounds that are potent and selective inhibitors of LRRK2 with a favorable pharmacokinetic profile and ability to cross the blood-brain barrier. Accordingly, the present invention relates to novel imidazo[4,5-c]quinoline compounds with LRRK2 inhibitory activity and the use of these compounds in the treatment of LRRK2 associated diseases such as neurodegenerative diseases including PD.

SUMMARY OF THE INVENTION

The present invention relates to compounds of formula I

where

R ¹ is selected from the group consisting of methyl, ethyl, cyclobutyl, cyclopentyl,

R ² is selected from the group consisting of 2,2-difluoropropyl,

and R ³ is selected from the group consisting of fluorine, chlorine, cyano, difluoromethyl and trifluoromethyl; or their pharmaceutically acceptable salt.

The present invention also relates to pharmaceutical compositions containing a pharmaceutically acceptable carrier and a compound of formula I or a pharmaceutically acceptable salt thereof, which are present in a therapeutically effective amount.

The present invention also relates to methods of treating a disorder or condition selected from Parkinson's disease (but also including other neurological diseases, which may include migraine; epilepsy; Alzheimer's disease; brain injury; stroke; cerebrovascular diseases (including cerebral arteriosclerosis, cerebral amyloid angiopathy, hereditary cerebral hemorrhage and cerebral hypoxia-ischemia); cognitive disorders (including amnesia, senile dementia, HIV-associated dementia, Alzheimer's disease, Huntington's disease, dementia with Lewy bodies, vascular dementia, drug-related dementia, tardive dyskinesia, myoclonus, dystonia, delirium, Pick's disease, Creutzfeldt-Jakob disease, HIV-induced disease, Gilles de la Tourette syndrome, epilepsy, muscle spasms and disorders associated with muscle spasticity or weakness, including tremors, and mild cognitive impairment); dementia (including spasticity, syndro m Down and fragile X syndrome); sleep disorders (including hypersomnia, circadian rhythm sleep disorder, insomnia, parasomnia and sleep deprivation) and psychiatric disorders such as anxiety (including acute stress disorder, generalized anxiety disorder, social anxiety disorder, panic disorder, post-traumatic stress disorder, agoraphobia and obsessive- compulsive disorder); simulation disorder (including hysterical twilight clouding of consciousness); impulse disorders (including compulsive gambling and intermittent explosive disorder); mood disorders (including bipolar I disorder, bipolar II disorder, mania, mixed affective disorder, major depressive episode, chronic depression, seasonal depression, psychotic depression, temporary depression, premenstrual syndrome (PMS), premenstrual dysphoric disorder (PMDD), and postpartum depression); psychomotor disorder; psychotic disorders (including schizophrenia, schizoaffective disorder, schizophreniform and delusional disorder); drug addiction (including drug addiction, alcoholism, amphetamine addiction, cocaine addiction, nicotine addiction and drug withdrawal syndrome); eating disorders (including anorexia, bulimia, compulsive overeating, hyperphagia, obesity, compulsive eating disorders and pagophagia); sexual dysfunction; urinary incontinence; neuronal damage (including eye damage, retinopathy or macular degeneration, tinnitus, hearing impairment and loss, and brain swelling) and psychiatric disorders in children (including attention deficit disorder, attention deficit hyperactivity disorder, conduct disorder, and autism) in a mammal , preferably human, comprising administering to the subject a therapeutically effective amount of a composition containing a compound of formula I or a pharmaceutically acceptable salt thereof.

It is to be understood that both the foregoing general description and the following detailed description are for the purpose of illustration and explanation only and do not limit the invention as claimed.

DETAILED DESCRIPTION OF THE INVENTION

The present invention can be more easily understood with reference to the following detailed description of exemplary embodiments of the invention and examples included herein.

It should be understood that the present invention is not limited to specific synthesis methods, which, of course, can be changed. It should also be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting. In this description and in the appended claims, reference will be made to a number of terms which, as will be defined below, have the following meanings:

As used herein, the singular form "one" may mean one or more than one. Used in the appended claims in combination with the word "comprising" the word "one" may mean one or more than one. As used herein, the term "other" may mean at least the second or subsequent.

The term "approximately" refers to a relative value, meaning an approximation of plus or minus 10% of the nominal value to which it refers, in one embodiment to plus or minus 5%, in another embodiment to plus or minus 2%. This level of approximation is applicable to the scope of the present invention, unless otherwise stated that the value requires a narrower range.

The term "treatment" as used herein, unless otherwise indicated, means reversing, alleviating symptoms, inhibiting progression, or preventing the disorder or condition to which such term applies, or one or more symptoms of such disorder or condition. The term "treatment" as used herein, unless otherwise indicated, refers to the treatment process as defined immediately above. The term "treatment" also includes adjuvant and neoadjuvant treatment of a subject.

The term "therapeutically effective amount" means an amount of a compound of the present invention that (I) treats or prevents a particular disease, condition, or disorder, (II) ameliorates, reduces, or eliminates one or more symptoms of a particular disease, condition, or disorder, or (III) prevents or delays the onset of one or more of the symptoms of a particular disease, condition, or disorder described herein. By "pharmaceutically acceptable" is meant that the substance or composition must be chemically and/or toxicologically compatible with the other ingredients that make up the composition and/or the mammal being treated with them.

As used herein, the terms "reaction inert solvent" and "inert solvent" refer to a solvent or mixture thereof that does not react with the starting compounds, reagents, intermediates, or products, thereby not adversely affecting the yield of the desired product.

The term "neurological" refers to the central nervous system. Treatment of neurological conditions refers to the treatment of a condition, disease, ailment, and so on, affecting the central nervous system ("CNS"). Such diseases can affect tissues in the peripheral as well as the central nervous system.

If substituents are described as "independently selected" from a group, then each substituent is selected independently of the other. Therefore, each substituent may be identical to or different from the other substituent(s).

As used herein, the terms "Formula I", "Formula I", "Formula (I)", or "Formula (I)" may be referred to as "compound(s) of the invention". Such terms also include all forms of the compound of formula I, including its hydrates, solvates, isomers, crystalline and non-crystalline forms, isomorphs, polymorphs and metabolites. For example, the compounds of the invention or their pharmaceutically acceptable salts may exist in unsolvated and solvated forms. When the solvent or water is strongly bound, the complex will have a well-defined stoichiometry regardless of the humidity. However, when the solvent or water is loosely bound, as in channel solvates and hygroscopic compounds, the water/solvent content will depend on humidity and drying conditions. In such cases, the absence of stoichiometry will be the norm.

The compounds of the invention may exist as clathrates or other complexes. Included within the scope of the invention are complexes such as clathrates, drug-host inclusion complexes, wherein the drug and host are present in stoichiometric or non-stoichiometric amounts. Also included are complexes of the compounds of the invention containing two or more organic and/or inorganic components, which may be present in stoichiometric or non-stoichiometric amounts. The resulting complexes may be ionized, partially ionized, or non-ionized. For a review of such complexes, see J. Pharm. Sci., 64(8), 1269-1288, Haleblian (August 1975).

), сплошной клиновидной линии (

) или пунктирной клиновидной линии (

). Использование сплошной линии для изображения связей с асимметрическими атомомами углерода предназначено для обозначения того, что включены все возможные стереоизомеры (например, соответствующие энантиомеры, рацемические смеси и так далее) по данному атому углерода. Использование либо сплошной, либо пунктирной клиновидной линии для изображения связей с асимметрическими атомами углерода предназначено для обозначения того, что должен быть включен только показанный стереоизомер. Возможно, что соединения формулы (I) могут содержать более одного асимметрического атома углерода. В таких соединениях использование сплошной линии для изображения связей с асимметрическими атомами углерода предназначено для обозначения того, что должны быть включены все возможные стереоизомеры. Например, если не оговорено особо, предполагается, что соединения формулы (I) могут существовать в виде энантиомеров и диастереомеров или в виде их рацематов и смесей. Использование сплошной линии для изображения связей с одним или более чем одним асимметрическим атомом углерода в соединении формулы (I) и использование сплошной или пунктирной клиновидной линии для изображения связей с другими асимметрическими атомами углерода в одном и том же соединении предназначено для обозначения того, что присутствует смесь диастереомеров.The compounds of the invention may have asymmetric carbon atoms. The carbon-carbon bonds of the compounds of the invention may be indicated here using a solid line (

), a solid wedge-shaped line (

) or dotted wedge line (

). The use of a solid line to depict bonds to asymmetric carbons is intended to indicate that all possible stereoisomers (eg, corresponding enantiomers, racemic mixtures, etc.) at that carbon atom are included. The use of either a solid or dashed wedge line to depict bonds to asymmetric carbons is intended to indicate that only the stereoisomer shown should be included. It is possible that the compounds of formula (I) may contain more than one asymmetric carbon atom. In such compounds, the use of a solid line to depict bonds to asymmetric carbon atoms is intended to indicate that all possible stereoisomers should be included. For example, unless otherwise indicated, it is contemplated that compounds of formula (I) may exist as enantiomers and diastereomers or as racemates and mixtures thereof. The use of a solid line to represent bonds to one or more asymmetric carbons in a compound of formula (I) and the use of a solid or dashed wedge to represent bonds to other asymmetric carbons in the same compound are intended to indicate that a mixture is present. diastereomers.

Stereoisomers of formula I include cis and trans isomers, optical isomers such as R and S enantiomers, diastereomers, geometric isomers, rotational isomers, conformational isomers and tautomers of the compounds of the invention, including compounds exhibiting more than one type of isomerism; and mixtures thereof (such as racemates and diastereomeric pairs). Also included are acid or base addition salts where the counterion is optically active, eg D-lactate or L-lysine, or racemic, eg DL-tartrate or DL-arginine.

When any racemate crystallizes, two different types of crystals are possible. The first type is the racemic compound (true racemate) mentioned above, where a single homogeneous crystal form is formed containing both enantiomers in equimolar amounts. The second type is a racemic mixture or conglomerate, where two forms of crystals are formed in equimolar amounts, each containing a separate enantiomer.

The chiral compounds of the invention (and their chiral precursors) can be obtained in an enantiomerically enriched form using chromatography, typically high pressure liquid chromatography (HPLC) or supercritical fluid chromatography (SFC), on a resin with an asymmetric stationary phase and with a mobile phase consisting of a hydrocarbon, typically heptane or hexane, containing 0 to 50% isopropanol, typically 2 to 20%, and 0 to 5% alkylamine, typically 0.1% diethylamine (DEA) or isopropylamine. Concentration of the eluent results in an enriched mixture.

Diastereomeric mixtures can be separated into individual diastereoisomers based on their physicochemical differences by methods well known to those skilled in the art, such as chromatography and/or fractional crystallization. Enantiomers can be separated by converting an enantiomeric mixture to a diastereomeric mixture by reacting with an appropriate optically active compound (e.g., a chiral auxiliary such as a chiral alcohol or Mosher acid chloride), separating the diastereoisomers, and converting (e.g., hydrolysis) the individual diastereoisomers to the corresponding pure enantiomers . Enantiomers can also be separated using a chiral HPLC column. Alternatively, specific stereoisomers can be synthesized using an optically active starting material, via asymmetric synthesis using optically active reagents, substrates, catalysts, or solvents, or by converting one stereoisomer to another via asymmetric transformation. The present invention includes tautomeric forms of the compounds of the invention. When structural isomers are capable of internal interconversion across a low energy barrier, tautomeric isomerism ("tautomerism") may exist. It can exist in the form of proton tautomerism in compounds of the invention containing, for example, an imino, keto or oxime group, or so-called valence tautomerism in compounds which contain an aromatic moiety. It follows that a single compound may exhibit more than one type of isomerism. Different ratios of tautomers in solid and liquid form depend on the different substituents in the molecule, as well as the particular crystallization technique used to isolate the compound.

The compounds of this invention may be used in the form of salts derived from inorganic or organic acids. Depending on the particular compound, the salt of the compound may benefit from one or more physical properties, such as improved pharmaceutical stability at varying temperatures and humidity, or desirable solubility in water or oil. In some cases, a salt of the compound may also be used to aid in the isolation, purification and/or redissolution of the compound.

When the salt is to be administered to a patient (as opposed to, for example, in vitro use), the salt is preferably pharmaceutically acceptable. The term "pharmaceutically acceptable salt" refers to a salt obtained by combining a compound of formula I with an acid whose anion or base the cation of which is generally considered to be suitable for human consumption. Pharmaceutically acceptable salts are particularly useful as products of the methods of the present invention due to their higher aqueous solubility compared to the parent compound. For use in medicine, the salts of the compounds of this invention are non-toxic "pharmaceutically acceptable salts." Salts, which are encompassed by the term "pharmaceutically acceptable salts", refer to non-toxic salts of the compounds of this invention, which are usually obtained by reacting a free base with a suitable organic or inorganic acid.

Suitable pharmaceutically acceptable acid addition salts of the compounds of the present invention, when possible, include those derived from inorganic acids such as hydrochloric, hydrobromic, hydrofluoric, boric, fluoroboric, phosphoric, metaphosphoric, nitric, carbonic, sulfonic and sulfuric acids, and organic acids such as as acetic, benzenesulfonic, benzoic, citric, ethanesulfonic, fumaric, gluconic, glycolic, isothionic, lactic, lactobionic, maleic, malic, methanesulfonic, trifluoromethanesulfonic, succinic, toluenesulfonic, tartaric and trifluoroacetic acids. Suitable organic acids generally include, for example, the classes of organic acids: aliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic, carboxylic, and sulfonic acids.

Specific examples of suitable organic acids include acetate, trifluoroacetate, formate salt, propionate, succinate, glycolate, gluconate, digluconate, lactate, malate, tartaric acid, citrate, ascorbate, glucuronate, maleate, fumarate, pyruvate, aspartate, glutamate, benzoate, anthranilic acid. , stearate, salicylate, para-hydroxybenzoate, phenylacetate, mandelate, embonate (pamoate), methanesulfonate, ethanesulfonate, benzenesulfonate, pantothenate, toluenesulfonate, 2-hydroxyethanesulfonate, sulfonate, cyclohexylaminosulfonate, β-hydroxybutyrate, galactarate, galacturonate, adipate, alginate, butyrate, camphorate, camphorsulfonate, cyclopentane propionate, dodecyl sulfate, glucoheptanoate, glycerophosphate, heptanoate, hexanoate, nicotinate, 2-naphthalenesulfonate, oxalate, palmoate, pectinate, 3-phenylpropionate, picrate, pivalate, thiocyanate and undecanoate.

In addition, when the compounds of the invention have an acid moiety, suitable pharmaceutically acceptable salts thereof may include alkali metal salts, ie sodium or potassium salts; alkaline earth metal salts, for example calcium or magnesium salts; and salts formed with suitable organic ligands, such as quaternary ammonium salts. In another embodiment, base salts are formed from bases that form non-toxic salts, including aluminum, arginine, benzathine, choline, diethylamine, diolamine, glycine, lysine, meglumine, olamine, tromethamine, and zinc salts.

Organic salts can be derived from salts of secondary, tertiary or quaternary amines such as tromethamine, diethylamine, N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N-methylglucamine) and procaine. Basic nitrogen-containing groups can be quaternized with agents such as lower alkyl(C ₁ -C ₆ ) halides (e.g. methyl, ethyl, propyl and butyl chlorides, bromides and iodides), dialkyl sulfates (i.e. dimethyl, diethyl, dibutyl - and diamyl sulfates), long chain halides (eg decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides), arylalkyl halides (eg benzyl and phenethyl bromides) and others.

In one embodiment, hemi-salts of acids and bases can also be formed, such as hemisulfate and hemi-calcium salts.

Also within the scope of the present invention are so-called "prodrugs" of the compound of the invention. Thus, certain derivatives of the compounds of the invention, which themselves may have little or no pharmacological activity, when administered to the body or applied to the body, can be converted to a compound of the invention having the desired activity, for example, by hydrolytic cleavage. Such derivatives are called "prodrugs". Additional information on the use of prodrugs can be found in "Pro-drugs as Novel Delivery Systems, Vol. 14, ACS Symposium Series (T. Higuchi and V. Stella) and "Bioreversible Carriers in Drug Design," Pergamon Press, 1987 (ed. E. B. Roche, American Pharmaceutical Association). for example in "Design of Prodrugs" by H. Bundgaard (Elsevier, 1985).

The present invention also includes isotopically labeled compounds which are identical to the compounds represented by formula I, with the only difference that one or more than one atom is replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number normally found in nature. . Examples of isotopes that can be included in the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine and chlorine, such as ² H, ³ H, ¹³ C, ¹¹ C, ¹⁴ C, ¹⁵ N , ¹⁸ O, ¹⁷ O, ³² P, ³⁵ S, ¹⁸ F, and ³⁶ Cl, respectively. The compounds of the present invention, their prodrugs and pharmaceutically acceptable salts of said compounds or said prodrugs which contain the aforementioned isotopes and/or other isotopes of other atoms are within the scope of this invention. Certain isotopically labeled compounds of the present invention, for example compounds in which radioactive isotopes such as ³ H and ¹⁴ C have been introduced, are useful in drug and/or substrate tissue distribution assays. Tritiated, ie ³ H, and carbon-14, ie ¹⁴ C, are especially preferred isotopes due to their ease of preparation and detection. In addition, substitution with heavier isotopes such as deuterium, i.e. ² H, may provide some therapeutic benefits resulting from higher metabolic stability, such as increased in vivo half-life or reduced dosage requirements, and therefore may in some cases be preferred. The isotopically labeled compounds of formula I of this invention and their prodrugs can generally be prepared by following the procedures disclosed in the Schemes and/or Examples for the preparation of compounds and intermediates below by substituting a non-isotopically labeled reagent with a readily available isotopically labeled reagent.

Typically, a compound of the invention is administered in an amount effective to treat the condition described herein. The compounds of the invention are administered by any suitable route in the form of a pharmaceutical composition adapted to such route and at a dose effective for the treatment indicated. Therapeutically effective doses of the compounds required to treat a progressive medical condition are readily determined by one of skill in the art using preclinical and clinical approaches known in the medical arts.

Pharmaceutical compositions for use in accordance with the present invention may be formulated in a conventional manner using one or more pharmaceutically acceptable carriers, including excipients and excipients which facilitate the conversion of the active compound into formulations that may have pharmaceutical uses. The appropriate drug depends on the chosen route of administration. Pharmaceutically acceptable excipients and carriers are generally known to those skilled in the art and are thus included in the present invention. Such excipients and carriers are described, for example, in "Remington's Pharmaceutical Sciences" Mack Pub. Co., New Jersey (1991). The formulations of the invention can be designed to be short acting, fast release, long acting and sustained release formulations. Thus, pharmaceutical preparations can also be formulated as controlled release or slow release preparations.

The pharmaceutical composition contains a compound of the invention or combination in an amount generally ranging from about 1% to about 75%, 80%, 85%, 90%, or even 95% (by weight) of the composition, usually ranging from about 1% , 2%, or 3% to about 50%, 60%, or 70%, more commonly ranging from about 1%, 2%, or 3% to less than 50%, such as up to about 25%, 30%, or 35%.

The compounds of the invention may be administered orally. Oral administration may include ingestion, such that the compound enters the gastrointestinal tract, or buccal or sublingual administration may be used, in which the compound enters the bloodstream directly from the oral cavity.

In another embodiment, the compounds of the invention may also be administered directly to the bloodstream, to a muscle, or to an internal organ. Suitable routes of parenteral administration include intravenous, intra-arterial, intraperitoneal, intrathecal, intraventricular, intraurethral, intrasternal, intracranial, intramuscular and subcutaneous. Suitable devices for parenteral administration include needle (including microneedle) injectors, needleless injectors and infusion techniques.

In another embodiment, the compounds of the invention may also be administered topically to the skin or mucous membranes, namely dermally or transdermally. In another embodiment, the compounds of the invention may also be administered intranasally or by inhalation. In another embodiment, the compounds of the invention may be administered rectally or vaginally. In another embodiment, the compounds of the invention may also be administered directly to the eye or ear.

The dosage regimen for compounds and/or compositions containing compounds is based on a variety of factors, including the type, age, body weight, sex, and medical condition of the patient; the severity of the condition; route of administration; and the activity of the particular compound used. Thus, the dosing regimen can vary within wide limits. Dosage levels on the order of about 0.01 mg to about 100 mg per kilogram of body weight per day are useful in the treatment of the above conditions. In one embodiment, the total daily dose of a compound of the invention (given in single or divided doses) is typically from about 0.01 to about 100 mg/kg. In another embodiment, the total daily dose of the compound of the invention is from about 0.1 to about 50 mg/kg, and in another embodiment from about 0.5 to about 30 mg/kg (i.e., mg of the compound of the invention per kg of body weight). In one embodiment, the dosage is from 0.01 to 10 mg/kg/day. In another embodiment, the dosage is from 0.1 to 1.0 mg/kg/day. Dosage unit compositions may contain such amounts or proportions thereof to constitute a daily dose. In many cases, compound administration is repeated multiple times per day (typically no more than 4 times). Multiple doses per day can usually be used to increase the total daily dose if required.

For oral administration, the compositions may be provided in the form of tablets containing from about 0.01 mg to about 500 mg of the active ingredient, or in another embodiment from about 1 mg to about 100 mg of the active ingredient. Doses for intravenous administration may vary from about 0.1 to about 10 mg/kg/minute at a constant infusion rate.

Suitable subjects of the present invention include mammalian subjects. Mammals of the present invention include dogs, cats, cattle, goats, horses, sheep, pigs, rodents, lagomorphs, primates, and the like, and encompasses, but is not limited to, mammals during fetal development. In one embodiment, humans are suitable subjects. Human subjects can be either gender-differentiated or at any stage of development.

In another embodiment, the invention includes the use of one or more compounds of the invention for the manufacture of a medicament for the treatment of the conditions described herein.

For the treatment of the above conditions, the compound of the invention may be administered as the compound itself. Alternatively, pharmaceutically acceptable salts are suitable for medical use due to their higher aqueous solubility compared to the parent compound.

In another embodiment, the present invention includes pharmaceutical compositions. Such pharmaceutical compositions contain a compound of the invention, presented together with a pharmaceutically acceptable carrier. The carrier may be a solid, liquid, or both, and may be formulated with the compound in a single dose formulation, such as a tablet, which may contain from 0.05% to 95% by weight of the active compounds. The compound of the invention can be combined with suitable polymers as carriers for targeted drug delivery. Other pharmacologically active substances may also be present.

The compounds of the present invention may be administered by any suitable route, preferably in the form of a pharmaceutical composition adapted to such route and at a dose effective for the intended treatment. Active compounds and compositions, for example, can be administered orally, rectally, parenterally or topically.

Oral administration of a solid dosage form may be presented, for example, in discrete units such as hard or soft capsules, pills, cachets, lozenges or tablets, each containing a predetermined amount of at least one compound of the present invention. In such solid dosage forms, a compound of the present invention or combination is admixed with at least one inert excipient, diluent or carrier. Suitable excipients, diluents or carriers include substances such as sodium citrate or dicalcium phosphate and/or (a) one or more fillers or modifying agents (e.g. microcrystalline cellulose (available as Avicel™ from FMC Corp.), starches, lactose, sucrose, mannitol, silicic acid, xylitol, sorbitol, dextrose, calcium hydrogen phosphate, dextrin, alpha-cyclodextrin, beta-cyclodextrin, polyethylene glycol, medium chain fatty acids, titanium oxide, magnesium oxide, alumina, and the like); (b) one or more binders (e.g., carboxymethylcellulose, methylcellulose a, hydroxypropylcellulose, hydroxypropylmethylcellulose, gelatin, gum arabic, ethylcellulose, polyvinyl alcohol, pullulan, pregelatinized starch, agar, tragacanth, alginates, gelatin, polyvinylpyrrolidone, sucrose, gum arabic and the like); (c) one or more humectants (eg glycerin and the like); (d) one or more disintegrants (e.g., agar-agar, calcium carbonate, potato or cassava starch, alginic acid, certain complex silicates, sodium carbonate, sodium lauryl sulfate, sodium starch glycolate (available as Explotab™ from Edward Mendell Co.) , cross-linked polyvinylpyrrolidone, croscarmellose sodium type A (available as Ac-Di-Sol™), polyacrylin-potassium (ion exchange resin) and the like); (e) one or more dissolution retarders (eg, paraffin and the like); (e) one or more absorption accelerators (eg, quaternary ammonium compounds and the like); (g) one or more wetting agents (eg, cetyl alcohol, glycerol monostearate, and the like); (h) one or more absorbent (eg kaolin, bentonite and the like); and/or (and) one or more lubricants (e.g., talc, calcium stearate, magnesium stearate, stearic acid, polyoxyl stearate, cetanol, talc, hydrogenated castor oil, sucrose fatty acid esters, dimethylpolysiloxane, microcrystalline wax, yellow bees wax, white beeswax, solid polyethylene glycols, sodium lauryl sulfate and the like). In the case of capsules and tablets, dosage forms may also contain buffering agents.

Solid compositions of a similar type can also be used as fillers in soft or hard filled gelatin capsules using excipients such as lactose or milk sugar, as well as high molecular weight polyethylene glycols and the like.

Solid dosage forms such as tablets, dragees, capsules and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the art. They may also contain opacifying agents and may also be formulated to release the compound of the present invention and/or the additional pharmaceutical agent in a delayed manner. Examples of encapsulating compositions that can be used are polymeric materials and waxes. The drug may also be in microencapsulated form, optionally with one or more of the aforementioned excipients.

For tablets, the active agent typically makes up less than 50% (by weight) of the formulation, such as less than about 10%, such as 5% or 2.5% by weight. Most of the preparation consists of fillers, diluents, disintegrants, lubricants and possibly flavoring agents. The composition of these excipients is well known in the art. Often fillers/diluents consist of a mixture of two or more of the following: microcrystalline cellulose, mannitol, lactose (all types) starch and dicalcium phosphate. Filler/diluent mixtures typically comprise less than 98% of the formulation and preferably less than 95%, eg 93.5%. Preferred disintegrants include Ac-Di-Sol™, Explotab™, starch and sodium lauryl sulfate. If a disintegrant is present, it typically makes up less than 10% of the formulation, or less than 5%, such as about 3%. A preferred lubricant is magnesium stearate. If a lubricant is present, it typically makes up less than 5% of the formulation, or less than 3%, such as about 1%.

Tablets can be prepared by standard tableting processes, such as direct compression, or wet or dry granulation, or melt granulation, melt solidification, and extrusion. Tablet cores may be single or multilayer and may be coated with suitable coatings known in the art.

In another embodiment, oral administration may be in a liquid dosage form. Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups and elixirs. In addition to the compound of the present invention or combination, liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents, solubilizers and emulsifiers, for example ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol , 1,3-butylene glycol, dimethylformamide, oils (e.g., cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, sesame oil, and the like), Miglyole.RTM (available from CONDEA Vista Co., Cranford, N.J. ), glycerin, tetrahydrofurfuryl alcohol, polyethylene glycols and sorbitan fatty acid esters or mixtures of these substances, and the like.

In addition to such inert diluents, the composition may also include excipients such as wetting agents, emulsifiers and suspending agents, sweeteners, flavors and flavors.

Liquid forms of the compounds of the invention or combinations for oral administration include solutions in which the active compound is completely dissolved. Examples of solvents include all pharmaceutically recognized solvents suitable for oral administration, in particular solvents in which the compounds of the invention exhibit good solubility, such as polyethylene glycol, polypropylene glycol, edible oils, and glyceryl and glyceride systems. Glyceryl and glyceride systems may include, for example, the following branded products (and corresponding generic products): Captex™ 355 EP (glyceryl tricaprylate/caprate from Abitec, Columbus Ohio), Crodamol™ GTC/C (medium chain triglyceride from Croda, Cowick Hall, UK) or Labrafac™ CC (medium chain triglycerides from Gattefosse), Captex™ 500P (glyceryl triacetate, i.e. triacetin from Abitec), Capmul™ MCM (medium chain mono- and diglycerides from Abitec), Migyol™ 812 (caprylic/capric triglyceride from Condea, Cranford NJ) , Migyol™ 829 (Caprylic/Capric/Succinic Triglyceride from Condea), Migyol™ 840 (Propylene Glycol Dicaprylate/Dicaprate from Condea), Labrafil™ M1944CS (Oleoyl Macrogol-6-Glycerides from Gattefosse), Peceol™ (Glyceryl Monooleate from Gattefosse), and Maisine™ 35 -1 (glyceryl monooleate from Gattefosse). Of particular interest are medium chain (approximately C ₈ -C ₁₀ ) triglyceride oils. These solvents often make up the majority of the composition, ie, more than about 50%, usually more than about 80%, such as about 95% or 99%. Adjuvants and excipients can also be included with solvents, mainly as taste-masking agents, flavoring agents and flavoring agents, antioxidants, stabilizers, density and viscosity modifiers, and solubilizers.

In addition to the compound of the present invention or combinations, the suspensions may further contain carriers such as suspending agents, for example ethoxylated isostearyl alcohols, polyoxyethylene sorbitan and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, or mixtures of these substances and the like.

In another embodiment, the present invention includes a parenteral dosage form. "Parenteral administration" includes, for example, subcutaneous injections, intravenous injections, intraperitoneal injections, intramuscular injections, intrasternal injections, and infusions. Injectable preparations (eg, sterile injectable aqueous or oily suspensions) may be prepared according to the known art using suitable dispersing, wetting and/or suspending agents. Compositions suitable for parenteral injection generally include pharmaceutically acceptable sterile aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, as well as sterile powders for preparing sterile injectable solutions or dispersions immediately prior to use. Examples of suitable aqueous and non-aqueous carriers or diluents (including solvents or excipients) include water, ethanol, polyols (propylene glycol, polyethylene glycol, glycerin, and the like), suitable mixtures thereof, triglycerides, including vegetable oils such as olive oil, and injectable organic compounds. esters such as ethyl oleate. A preferred carrier is the ester of caprylic/capric acid with glycerol or propylene glycol under the tradename Miglyol.RTM (e.g. Miglyol.RTM 812, Miglyol.RTM 829, Miglyol.RTM 840) available from Condea Vista Co., Cranford, N.J. Proper fluidity can be maintained, for example, by the use of coating agents such as lecithin, by maintaining the required particle size in the case of dispersions, or by the use of surfactants.

These compositions for parenteral injection may also contain excipients such as preservatives, wetting agents, emulsifiers and dispersing agents. Prevention of contamination of the compositions by microorganisms can be ensured by the inclusion of various antibacterial and antifungal agents, for example parabens, chlorobutanol, phenol, sorbic acid, and the like. It may also be desirable to include isotonic agents such as sugars, sodium chloride and the like. Prolonged absorption of injectable pharmaceutical compositions can be achieved through the use of agents that delay absorption, such as aluminum monostearate and gelatin. In another embodiment, the present invention includes a topical dosage form. "Topical administration" includes, for example, transdermal administration, such as via transdermal patches or iontophoresis devices, intraocular administration, or intranasal or inhalation administration. Compositions for topical administration also include, for example, topical gels, sprays, ointments and creams. Topical administration may include a compound that enhances the absorption or penetration of the active ingredient through the skin or other affected areas. When the compounds of this invention are administered via a transdermal device, the administration is carried out using a patch with either a reservoir and a porous membrane or a variety of solid matrices. Typical formulations for this purpose include gels, hydrogels, lotions, solutions, creams, ointments, powders, dressings, foams, films, skin patches, cachets, implants, sponges, fibers, bandages, and microemulsions. Liposomes may also be used. Typical carriers include alcohol, water, mineral oil, liquid petrolatum, white petrolatum, glycerin, polyethylene glycol, and propylene glycol. Penetration enhancers may be included; see for example J. Pharm. Sci., 88 (10), 955-958, Finnin and Morgan (October 1999).

Formulations suitable for topical administration to the eye include, for example, eye drops wherein the compound of this invention is dissolved or suspended in a suitable carrier. A topical preparation suitable for ophthalmic or otic administration may be in the form of drops of a micronized suspension or solution in pH-adjusted isotonic sterile saline. Other formulations suitable for ocular and otic administration include ointments, biodegradable (eg, absorbable gel sponges, collagen) and non-biodegradable (eg, silicone) implants, cachets, lenses, and particulate or vesicular systems such as niosomes or liposomes. A polymer such as cross-linked polyacrylic acid, polyvinyl alcohol, hyaluronic acid, a cellulose polymer such as (hydroxypropyl)methylcellulose, hydroxyethylcellulose or methylcellulose, or a heteropolysaccharide polymer such as gellan gum may be included along with a preservative such as benzalkonium chloride. Such drugs can also be delivered by iontophoresis.

For intranasal or inhalation administration, the active compounds of the invention are suitably delivered in the form of a solution or suspension from an atomizer which is compressed or pumped by the patient, or in the form of an aerosol spray from a pressurized container or nebulizer using a suitable propellant. Formulations suitable for intranasal administration are usually administered in dry powder form (either singly, as a mixture, eg dry-mixed with lactose, or as a particle of a mixed component, eg mixed with phospholipids such as phosphatidylcholine) from a dry powder inhaler or as an aerosol spray from a pressurized container, pump, spray, nebulizer (preferably a nebulizer using electrohydrodynamics to produce a fine mist) or nebulizer, with or without the use of a suitable propellant such as 1,1,1,2-tetrafluoroethane or 1,1,1,2-tetrafluoroethane or 1, 1,1,2,3,3,3-heptafluoropropane. For intranasal use, the powder may contain a bioadhesive agent such as chitosan or cyclodextrin.

In another embodiment, the present invention includes a rectal or vaginal dosage form. Such a rectal dosage form may be presented as, for example, a suppository. Cocoa butter, polyethylene glycol and suppository wax are traditional suppository bases, but various alternatives can be used if desired. These bases are solid at normal room temperature but liquid at body temperature and will therefore melt in the rectum or vaginal cavity to release the active ingredient(s).

Many of the compounds of the present invention are sparingly soluble in water, eg having a solubility of less than about 1 μg/mL. Therefore, liquid formulations in solubilizing non-aqueous solvents such as the medium chain triglyceride oils discussed above are the preferred dosage forms for these compounds.

Solid amorphous dispersions, including those formed by spray drying, are also a preferred dosage form for the slightly soluble compounds of the invention. By "solid amorphous dispersion" is meant a solid material in which at least a portion of the slightly soluble compound is in amorphous form and dispersed in a water-soluble polymer. By "amorphous" is meant that the slightly soluble compound is not crystalline. By "crystalline" is meant that the compound exhibits long-range order in three dimensions of at least 100 repeat units in each dimension. Thus, the term "amorphous" is intended to include not only a substance that is essentially without order, but also a substance that may have some small degree of order, but order in less than three dimensions and/or only in the short range. distances. An amorphous substance can be characterized using techniques known in the art such as powder X-ray diffraction (PXRD), crystallography, NMR (nuclear magnetic resonance) of a solid, or by thermal methods such as differential scanning calorimetry (DSC) .

Preferably, at least a major portion (ie, at least about 60% (wt.)) of the slightly soluble compound in the solid amorphous dispersion is amorphous. The compound may be in a solid amorphous dispersion in relatively pure amorphous domains or regions, as a solid solution of the compound homogeneously distributed in the polymer, or any combination of these states or those states that are intermediate between them. Preferably, the solid amorphous dispersion is substantially homogeneous so that the amorphous compound is as homogeneously dispersed as possible in the polymer. The term "substantially homogeneous" as used herein means that the portion of the compound that is present in relatively pure amorphous domains or regions in the solid amorphous dispersion is relatively small, on the order of less than about 20% by weight, and preferably less than about 10%. (wt.) of the total amount of the drug.

Water-soluble polymers suitable for use in solid amorphous dispersions should be inert in that they do not chemically react with the poorly soluble compound in an undesirable manner, are pharmaceutically acceptable, and have at least some solubility in aqueous solution at physiological pH ( e.g. 1-8). The polymer may be neutral or ionizable and should have a water solubility of at least 0.1 mg/mL over at least part of the pH range 1-8.

Water-soluble polymers suitable for use in the present invention may be cellulosic or non-cellulosic. The polymers may be neutral or ionizable in aqueous solution. Of these, ionizable and cellulosic polymers are preferred, with ionizable cellulosic polymers being more preferred.

Representative water-soluble polymers include hydroxypropyl methylcellulose acetate succinate (HPMCAS), hydroxypropyl methylcellulose (HPMC), hydroxypropyl methylcellulose phthalate (HPMCP), carboxymethyl ethyl cellulose (CMEC), cellulose acetate phthalate (CAP), cellulose acetate trimellitate (CAT), polyvinyl pyrrolidone (PVP), hydroxypropyl cellulose (HPC) , methylcellulose (MS), ethylene oxide-propylene oxide block copolymers (PEO/PPO, also known as poloxamers), and mixtures thereof. Particularly preferred polymers include HPMCAS, HPMC, HPMCP, CMEC, CAP, CAT, PVP, poloxamers and mixtures thereof. HPMCAS is most preferred. See European Patent Application Publication No. 0901786 A2, the disclosure of which is incorporated herein by reference.

Solid amorphous dispersions can be obtained by any method of formation of solid amorphous dispersions, which results in at least the majority (at least 60%) of the slightly soluble compound is in the amorphous state. Such methods include mechanical, thermal and dissolution methods. Typical mechanical methods include milling and extrusion; melting methods include high temperature melting, solvent-modified melting, and melt solidification methods; and dissolution methods include precipitation with a non-solvent, spray coating, and spray drying. See, for example, the following US Pat. No. 5340591 and 4673564, which describe the formation of dispersions by grinding methods; and Nos. 5,707,646 and 4,894,235, which describe the formation of dispersions by melt solidification methods. In a preferred process, a solid amorphous dispersion is formed by spray drying as disclosed in European Patent Application Publication No. 0901786 A2. In this method, the compound and the polymer are dissolved in a solvent such as acetone or methanol, and then the solvent is quickly removed from the solution by spray drying to form a solid amorphous dispersion. Can be obtained solid amorphous dispersion containing up to about 99% (wt.) compounds, for example 1% (wt.), 5% (wt.), 10% (wt.), 25% (wt.), 50% ( wt.), 75% (wt.), 95% (wt.) or 98% (wt.), as required.

The solid dispersion may be used as a stand-alone dosage form or may serve as a manufacturing-use-product (PIP) product for other dosage forms such as capsules, tablets, solutions or suspensions. An example of an aqueous suspension is an aqueous suspension of a spray-dried compound/HPMCAS-HF (1:1, w/w) dispersion containing 2.5 mg/ml of the compound in 2% polysorbate-80. Solid dispersions for use in a tablet or capsule are usually mixed with other excipients or adjuvants commonly found in such dosage forms. For example, a typical capsule filler contains a spray-dried compound/HPMCAS-HF (2:1, w/w) dispersion (60%), lactose (free flowing) (15%), microcrystalline cellulose (e.g., Avicel. sup. Ph -102) (15.8%), sodium starch (7%), sodium lauryl sulfate (2%) and magnesium stearate (1%).

HPMCAS polymers are available in low, medium and high grades such as Aqoa.sup.(R)-LF, Aqoat.sup.(R)-MF, and Aqoat.sup.(R)-HF, respectively, from Shin-Etsu Chemical Co. , LTD, Tokyo, Japan. The higher grades MF and HF are generally preferred.

Other carrier substances and routes of administration known in the pharmaceutical art may also be used. Pharmaceutical compositions of the invention may be prepared by any of the well known pharmaceutical techniques such as efficient preparation and administration techniques. The foregoing discussions regarding effective formulations and administration techniques are well known in the art and are described in standard guidelines. Drug formulations are discussed in, for example, Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania, 1975; Liberman et al., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, NY, 1980; and ^Kibbe et al., Eds., Handbook of Pharmaceutical Excipients (3rd Ed.), American Pharmaceutical Association, Washington, 1999.

The compounds of the present invention may be used alone or in combination with other therapeutic agents in the treatment of various conditions and diseases. The compound(s) of the present invention and the other(s) therapeutic(s) agent(s) may be administered simultaneously (either in the same dosage form or in separate dosage forms) or sequentially.

Two or more compounds may be administered simultaneously, in parallel, or sequentially. Additionally, simultaneous administration can be accomplished by mixing the compounds prior to administration, or by administering the compounds at the same time but at different anatomical sites or using different routes of administration.

The phrases "parallel administration", "co-administration", "simultaneous administration" and "administered simultaneously" mean that the compounds are administered in combination. The present invention includes the use of a combination of an LRRK2 inhibitor compound as provided in Formula I and one or more additional pharmaceutically active agent(s). If a combination of active agents is administered, then they may be administered sequentially or simultaneously in separate dosage forms or combined into one dosage form. Accordingly, the present invention also includes pharmaceutical compositions containing an amount of: (a) a first agent containing a compound of formula I or a pharmaceutically acceptable salt of this compound; (b) a second pharmaceutically active agent; and (c) a pharmaceutically acceptable carrier, excipient or diluent.

Various pharmaceutically active agents may be selected for use in combination with the compounds of formula (I) depending on the disease, disorder or condition requiring treatment. For example, a pharmaceutical composition for use in the treatment of Parkinson's disease may contain a compound of Formula I, or a pharmaceutically acceptable salt thereof, together with another agent such as dopamine (levodopa, either alone or with a DOPA decarboxylase inhibitor), a monoamine oxidase (MAO) inhibitor, a catechol inhibitor. -O-methyltransferase (COMT) or an anticholinergic agent, or any combination thereof. Particularly preferred agents for combination with compounds of formula (I) for use in the treatment of Parkinson's disease include levodopa, carbidopa, tolcapone, entacapone, selegiline, benztropine and trihexyphenidyl, or any combination thereof. Pharmaceutically active agents which may be used in combination with compounds of formula (I) and compositions thereof include, without limitation:

(I) levodopa (or its methyl or ethyl ester) alone or in combination with a DOPA decarboxylase inhibitor (eg, carbidopa (SINEMET, CARBILEV, PARCOPA), benserazide (MADOPAR), α-methyldopa, monofluoromethyldopa, difluoromethyldopa, brocresin, or meta -hydroxybenzylhydrazine);

(II) anticholinergic agents such as amitriptyline (ELAVIL, ENDEP), butriptyline, benztropine mesylate (COGENTIN), trihexyphenidyl (ARTANE), diphenhydramine (BENADRYL), orphenadrine (NORFLEX), hyoscyamine, atropine (ATROPEN), scopolamine (TRANSDERM-SCOP) ), scopolamine methyl bromide (PARMINE), dicycloverine (BENTYL, BYCLOMINE, DIBENT, DILOMINE), tolterodine (DETROL), oxybutynin (DITROPAN, LYRINEL XL, OXYTROL), pentienate bromide, propantheline (PRO-BANTHINE), cyclizine, imipramine hydrochloride (TOFRANIL ), imipramine maleate (SURMONTIL), lofepramine, desipramine (NORPRAMIN), doxepin (SINEQUAN, ZONALON), trimipramine (SURMONTIL), and glycopyrrolate (ROBINUL);

(III) catechol-O-methyltransferase (COMT) inhibitors such as nitecapone, tolcapone (TASMAR), entacapone (COMTAN) and tropolone;

(iv) monoamine oxidase (MAO) inhibitors such as selegiline (EMSAM), selegiline hydrochloride (1-deprenyl, ELDEPRYL, ZELAPAR), dimethylselegiline, brofaromine, phenelzine (NARDIL), tranylcypromine (PARNATE), moclobemide (AURORIX, MANERIX), befloxatone , safinamide, isocarboxazid (MARPLAN), nialamide (NIAMID), rasagiline (AZILECT), iproniazid (MARSILID, IPROZID, IPRONID), iproclozide, toloxatone (HUMORYL, PERENUM), bifemelan, deoxypeganine, harmine (also known as telepatin or banasterin), harmaline, linezolid (ZYVOX, ZYVOXID) and pargyline (EUDATIN, SUPIRDYL);

(V) acetylcholinesterase inhibitors such as donepezil hydrochloride (ARICEPT®, MEMAC), physostigmine salicylate (ANTILIRIUM®), physostigmine sulfate (ESERINE), ganstigmine, rivastigmine (EXELON®), ladostigil, NP-0361, galantamine hydrobromide (RAZADYNE®, REMINYL®, NIVALIN®), tacrine (COGNEX®), tolserine, memoquin, huperzine A (HUP-A; Neuro-Hitech), fenserine, bisnorcymserine (also known as BNC), and INM-176;

(VI) amyloid-β (or fragments thereof), eg Aβ _1-15 -conjugated to a universal HLA DR-binding epitope (PADRE®), ACC-001 (Elan/Wyeth) and Affitope;

(vii) antibodies to amyloid-β (or fragments thereof) such as ponezumab, solanezumab, bapineuzumab (also known as AAB-001), AAB-002 (Wyeth/Elan), gantenerumab, intravenous Ig (immunoglobulin) (GAMMAGARD®) , LY2062430 (humanized m266; Lilly) and described in International Application Publication Nos. WO 04/032868, WO 05/025616, WO 06/036291, WO06/069081, WO 06/118959, US Patent Application Publication No. US 2003 /0073655, US 2004/0192898, US 2005/0048049, US 2005/0019328, European Patent Publication Nos. EP 0994728 and 1257584 and US Patent No. 5750349;

(VIII) amyloid-lowering or -inhibiting agents (including those that reduce amyloid production, accumulation and fibrillation) such as eprodisate, celecoxib, lovastatin, anapsos, colostrinin, pioglitazone, clioquinol (also known as PBT1), PBT2 (Prana Biotechnology ), flurbiprofen (ANSAID®, FROBEN®) and its R-enantiomer tarenflurbil (FLURIZAN®), nitroflurbiprofen, fenoprofen (FENOPRON, NALFON®), ibuprofen (ADVIL®, MOTRIN®, NUROFEN®), ibuprofen lysinate, meclofenamic acid, meclofenamate sodium (MECLOMEN®), indomethacin (INDOCIN®), diclofenac sodium (VOLTAREN®), diclofenac potassium, sulindac (CLINORIL®), sulindac sulfide, diflunisal (DOLOBID®), naproxen (NAPROSYN®), naproxen sodium (ANAPROX®, ALEVE ®), Insulin Degrading Enzyme (also known as Insulisin), Gingko Biloba Extract EGb-761 (ROKAN®, TEBONIN®), Tramiprosate (CEREBRIL®, ALZHEMED®), KIACTA®), Neprilysin (also known as Neutral Endopeptidase (NEP )), scillo-inositol (also known as scillitol), atorv astatine (LIPITOR®), simvastatin (ZOCOR®), ibutamorena mesylate, BACE (beta-secretase 1) inhibitors such as LY450139 (Lilly), BMS-782450, GSK-188909; gamma-secretase modulators and inhibitors such as ELND-007, BMS-708163 (Avagacestat) and DSP8658 (Dainippon); and RAGE inhibitors (receptor for advanced glycation end products) such as TTP488 (Transtech) and TTP4000 (Transtech) as well as those described in US Pat. No. 7,285,293, including PTI-777;

(IX) alpha-adrenergic receptor agonists and beta-adrenergic receptor blocking agents (beta-blockers); anticholinergic agents; anticonvulsants; antipsychotic agents; calcium channel blockers; catechol-O-methyltransferase (COMT) inhibitors; stimulants of the central nervous system; corticosteroids; agonists and antagonists of dopamine receptors; dopamine reuptake inhibitors; gamma-aminobutyric acid (GABA) receptor agonists; immunosuppressive agents; interferons; muscarinic receptor agonists; neuroprotective drugs; nicotinic receptor agonists; norepinephrine (norepinephrine) reuptake inhibitors; quinolines and trophic factors;

(X) histamine 3 (H3) antagonists such as PF-3654746 and described in US Patent Application Publication Nos. US 2008-0176925; International Patent Publication Nos. WO 2006/136924, WO 2007/063385, WO 2007/069053, WO 2007/088450, WO 2007/099423, WO 2007/105053, WO 2007/138431 and WO 2007/088462; and US Pat. No. 7,115,600;

(XI) N-methyl-D-aspartate (NMDA) receptor antagonists such as memantine (NAMENDA, AXURA, EBIXA), amantadine (SYMMETREL), acamprosate (CAMPRAL), besonprodil, ketamine (KETALAR), delucemine, dexanabinol, dexefaroxan, dextromethorphan, dextrophan, traxoprodil, CP-283097, himantane, idantadol, ipenoxazone, L-701252 (Merck), lancicemin, levorphanol (DROMORAN), methadone, (DOLOPHINE), neramexane, perzinfotel, phencyclidine, tianeptine (STABLON), dizocilpine (also known as MK-801), ibogaine, voacangin, tiletamine, riluzole (RILUTEK), aptiganel (CERESTAT), gavestinel, and remacimide;

(XII) phosphodiesterase (PDE) inhibitors, including (a) PDE1 inhibitors; (b) PDE2 inhibitors; (c) PDE3 inhibitors; (d) PDE4 inhibitors; (e) PDE5 inhibitors; (e) PDE9 inhibitors (e.g., PF-04447943, BAY 73-6691 (Bayer AG) and those described in US Patent Application Publication Nos. US 2003/0195205, US 2004/0220186, US 2006/0111372, US 2006/0106035 and USSN 12/118062 (filed May 9, 2008)); and (g) PDE10 inhibitors such as 2-({4-[1-methyl-4-(pyridin-4-yl)-1H-pyrazol-3-yl]phenoxy}methyl)quinoline (PF-2545920);

(XIII) serotonin (5-hydroxytryptamine) 1A (5-HT _1A ) receptor antagonists such as spiperone, levo-pindolol, lecozotan;

(XIV) serotonin (5-hydroxytryptamine) 2C (5-HT _2c ) receptor agonists such as vabicaserin and cyclonapine; serotonin (5-hydroxytryptamine) 4 (5-HT ₄ ) receptor agonists/antagonists such as PRX-03140 (Epix) and PF-04995274;

(XV) serotonin (5-hydroxytryptamine) receptor antagonists 3C (5-HT _3c ) such as ondansetron (Zofran);

(XVI) Serotonin (5-hydroxytryptamine) 6 (5-HT ₆ ) receptor antagonists such as mianserin (TOLVON, BOLVIDON, NORVAL), methiothepin (also known as methitepin), ritanserin, SB-271046, SB-742457 (GlaxoSmithKline) , Lu AE58054 (Lundbeck A/S), SAM-760 and PRX-07034 (Epix);

(XVII) serotonin (5-HT) reuptake inhibitors such as alaproclate, citalopram (CELEXA, CIPRAMIL), escitalopram (LEXAPRO, CIPRALEX), clomipramine (ANAFRANIL), duloxetine (CYMBALTA), femoxetine (MALEXIL), fenfluramine (PONDIMIN) , norfenfluramine, fluoxetine (PROZAC), fluvoxamine (LUVOX), indalpine, milnacipran (IXEL), paroxetine (PAXIL, SEROXAT), sertraline (ZOLOFT, LUSTRAL), trazodone (DESYREL, MOLIPAXIN), venlafaxine (EFFEXOR), zimelidine (NORMUD, ZELMID), bicifadine, dezvenlafaxine (PRISTIQ), brazofenzine, vilazodone, cariprazine, and tesofensine;

(XVIII) glycine transporter-1 inhibitors such as paliflutin, ORG-25935 and ORG-26041; and mGluR modulators such as AFQ-059 and amantidine;

(XIX) AMPA-type glutamate receptor modulators such as perampanel, mibampator, selurampanel, GSK-729327 and N-{(3S,4S)-4-[4-(5-cyanothiophen-2-yl)phenoxy]tetrahydrofuran- 3-yl}propan-2-sulfonamide;

(XX) P450 inhibitors such as ritonavir;

(XXI) tau targeted therapy targets such as davunetide;

etc.

The present invention further includes kits that are suitable for use in the practiced treatments described above. In one embodiment, the kit contains a first dosage form containing one or more compounds of the present invention and a dosage form container, in amounts sufficient to carry out the methods of the present invention.

In another embodiment, the kit of the present invention contains one or more compounds of the invention.

In one embodiment, the compound of the present invention is:

[(2S,4R)-4-(8-chloro-2-ethyl-1H-imidazo[4,5-c]quinolin-1-yl)tetrahydro-2H-pyran-2-yl]acetonitrile;

[(2R,4S)-4-(8-chloro-2-ethyl-1H-imidazo[4,5-c]quinolin-1-yl)tetrahydro-2H-pyran-2-yl]acetonitrile;

1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(4-methyl-2H-1,2,3-triazol-2-yl)methyl]-1H-imidazo[4,5 -c]quinoline-8-carbonitrile, ENT 1 (enantiomer 1);

1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(4-methyl-2H-1,2,3-triazol-2-yl)methyl]-1H-imidazo[4,5 -c]quinoline-8-carbonitrile, ENT 2 (enantiomer 2);

8-chloro-1-[(4S)-3,3-difluorotetrahydro-2H-pyran-4-yl]-2-[(5-methyl-1,2-oxazol-3-yl)methyl]-1H-imidazo [4,5-c]quinoline;

2-[(6-methylpyrimidin-4-yl)methyl]-1-[(3R)-1-methylpyrrolidin-3-yl]-1H-imidazo[4,5-c]quinolin-8-carbonitrile;

8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-[(5-methylpyrazin-2-yl)methyl]-1H-imidazo[4,5-c]quinoline, ENT 1 (enantiomer 1);

8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-[(5-methylpyrazin-2-yl)methyl]-1H-imidazo[4,5-c]quinoline, ENT 2 (enantiomer 2);

1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-2-[(1-methyl-1H-1,2,3-triazol-4-yl)methyl]-8-( trifluoromethyl)-1H-imidazo[4,5-c]quinoline;

[cis-4-(8-chloro-2-cyclobutyl-1H-imidazo[4,5-c]quinolin-1-yl)tetrahydro-2H-pyran-2-yl]acetonitrile, ENT 1 (enantiomer 1);

[cis-4-(8-chloro-2-cyclobutyl-1H-imidazo[4,5-c]quinolin-1-yl)tetrahydro-2H-pyran-2-yl]acetonitrile, ENT 2 (enantiomer 2);

8-(difluoromethyl)-2-[(4-methoxy-1H-pyrazol-1-yl)methyl]-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo [4,5-c]quinoline;

8-(difluoromethyl)-2-[(5-methylpyrazin-2-yl)methyl]-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[4,5 -c]quinoline;

{8-chloro-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[4,5-c]quinolin-2-yl}(5-methylpyrazin-2- yl) methanol, DIAST 1 (diastereomer 1);

{8-chloro-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[4,5-c]quinolin-2-yl}(5-methylpyrazin-2- yl) methanol, DIAST 2 (diastereomer 2);

1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-8-fluoro-2-(1H-1,2,4-triazol-1-ylmethyl)-1H-imidazo[4,5-с] quinoline, ENT 1 (enantiomer 1);

1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-8-fluoro-2-(1H-1,2,4-triazol-1-ylmethyl)-1H-imidazo[4,5-с] quinoline, ENT 2 (enantiomer 2);

1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-8-fluoro-2-[(4-methyl-1H-1,2,3-triazol-1-yl)methyl]-1H-imidazo [4,5-c]quinoline, ENT 1 (enantiomer 1);

1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-8-fluoro-2-[(4-methyl-1H-1,2,3-triazol-1-yl)methyl]-1H-imidazo [4,5-c]quinoline, ENT 2 (enantiomer 2);

1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-8-fluoro-2-[(5-methylpyrazin-2-yl)methyl]-1H-imidazo[4,5-c]quinoline, ENT 1 (enantiomer 1);

1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-8-fluoro-2-[(5-methylpyrazin-2-yl)methyl]-1H-imidazo[4,5-c]quinoline, ENT 2 (enantiomer 2);

8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-{[4-(methoxymethyl)-1H-1,2,3-triazol-1-yl]methyl}-1H -imidazo[4,5-c]quinoline, ENT 1 (enantiomer 1);

8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-{[4-(methoxymethyl)-1H-1,2,3-triazol-1-yl]methyl}-1H -imidazo[4,5-c]quinoline, ENT 2 (enantiomer 2);

8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-(1Н-1,2,4-triazol-1-ylmethyl)-1Н-imidazo[4,5-с] quinoline, ENT 1 (enantiomer 1);

8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-(1Н-1,2,4-triazol-1-ylmethyl)-1Н-imidazo[4,5-с] quinoline, ENT 2 (enantiomer 2);

8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-[(4-methoxy-1H-pyrazol-1-yl)methyl]-1H-imidazo[4,5-c ]quinoline, ENT 1 (enantiomer 1);

8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-[(4-methoxy-1H-pyrazol-1-yl)methyl]-1H-imidazo[4,5-c ]quinoline, ENT 2 (enantiomer 2);

8-fluoro-2-[(2-methylimidazo[2,1-b][1,3,4]thiadiazol-6-yl)methyl]-1-[(2R,4R)-2-methyltetrahydro-2H-pyran -4-yl]-1H-imidazo[4,5-c]quinoline;

2-[(5-methylpyrazin-2-yl)methyl]-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-8-(trifluoromethyl)-1H-imidazo[4,5 -c]quinoline;

2-cyclopentyl-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[4,5-c]quinoline-8-carbonitrile;

[cis-4-(8-chloro-2-methyl-1H-imidazo[4,5-c]quinolin-1-yl)tetrahydro-2H-pyran-2-yl]acetonitrile, ENT 1 (enantiomer 1);

1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(5-methyl-1,2,4-oxadiazol-3-yl)methyl]-1H-imidazo[4,5-c ]quinoline-8-carbonitrile, ENT 1 (enantiomer 1);

2-[(5-methylpyrazin-2-yl)methyl]-1-[(3R)-1-methylpyrrolidin-3-yl]-1H-imidazo[4,5-c]quinolin-8-carbonitrile;

1-[(3R)-1-methylpyrrolidin-3-yl]-2-[(5-methyl-2H-tetrazol-2-yl)methyl]-1H-imidazo[4,5-c]quinolin-8-carbonitrile ;

2-[(3-methyl-1,2-oxazol-5-yl)methyl]-1-[(3R)-1-methylpyrrolidin-3-yl]-1H-imidazo[4,5-c]quinoline-8 -carbonitrile;

2-[(4-Methoxy-1H-pyrazol-1-yl)methyl]-1-[(3R)-1-methylpyrrolidin-3-yl]-1H-imidazo[4,5-c]quinolin-8-carbonitrile ;

1-[(3R)-1-methylpyrrolidin-3-yl]-2-[(5-methyl-1,3,4-thiadiazol-2-yl)methyl]-1H-imidazo[4,5-c]quinoline -8-carbonitrile;

2-[(5-methyl-1,3-oxazol-2-yl)methyl]-1-[(3R)-1-methylpyrrolidin-3-yl]-1H-imidazo[4,5-c]quinoline-8 -carbonitrile;

1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-2-{[5-(trifluoromethyl)pyrazin-2-yl]methyl}-1H-imidazo [4,5-c] quinoline-8-carbonitrile;

8-chloro-2-[(6-methylpyrimidin-4-yl)methyl]-1-[(3R)-1-methylpyrrolidin-3-yl]-1H-imidazo[4,5-c]quinoline;

2-[(4-methoxy-1H-pyrazol-1-yl)methyl]-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-8-(trifluoromethyl)-1H-imidazo [4,5-c]quinoline;

8-chloro-2-[(5-methyl-1,2,4-oxadiazol-3-yl)methyl]-1-[(3R)-1-methylpyrrolidin-3-yl]-1H-imidazo[4,5 -c]quinoline;

8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-(1H-1,2,4-triazol-1-ylmethyl)-1H-imidazo[4,5-с] quinoline;

8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-{[4-(methoxymethyl)-1H-1,2,3-triazol-1-yl]methyl}-1H -imidazo[4,5-c]quinoline;

8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(5-methyl-1,2,4-oxadiazol-3-yl)methyl]-1H-imidazo[4 ,5-c]quinoline;

8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-[(4-methyl-1H-1,2,3-triazol-1-yl)methyl]-1H-imidazo [4,5-c]quinoline, ENT 1 (enantiomer 1);

8-chloro-2-[(4-cyclopropyl-1H-1,2,3-triazol-1-yl)methyl]-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-1H-imidazo [4,5-c]quinoline, ENT 2 (enantiomer 2);

8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(4-methyl-1H-1,2,3-triazol-1-yl)methyl]-1H-imidazo [4,5-c]quinoline, ENT 2 (enantiomer 2);

8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(5-methyl-2H-tetrazol-2-yl)methyl]-1H-imidazo[4,5-c ]quinoline, ENT 1 (enantiomer 1);

8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(5-methylpyrazin-2-yl)methyl]-1H-imidazo[4,5-c]quinoline, ENT 1 (enantiomer 1);

8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(5-methyl-1,2-oxazol-3-yl)methyl]-1H-imidazo[4,5 -c]quinoline, ENT 1 (enantiomer 1);

8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(5-methyl-1,2-oxazol-3-yl)methyl]-1H-imidazo[4,5 -c]quinoline, ENT 2 (enantiomer 2);

1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-8-fluoro-2-{[4-(methoxymethyl)-1H-1,2,3-triazol-1-yl]methyl}-1H -imidazo[4,5-c]quinoline, ENT 2 (enantiomer 2);

8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-(1H-1,2,4-triazol-1-ylmethyl)-1H-imidazo[4,5-c] quinoline, ENT 2 (enantiomer 2);

8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-{[4-(methoxymethyl)-1H-1,2,3-triazol-1-yl]methyl}-1H -imidazo[4,5-c]quinoline, ENT 1 (enantiomer 1);

2-[(5-methyl-1,2,4-oxadiazol-3-yl)methyl]-1-[(3R)-1-(2,2,2-trifluoroethyl)pyrrolidin-3-yl]-1H- imidazo[4,5-c]quinoline-8-carbonitrile;

2-[(4-methoxy-1H-pyrazol-1-yl)methyl]-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[4,5-с ]quinoline-8-carbonitrile;

8-chloro-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-2-(1,3-thiazol-2-ylmethyl)-1H-imidazo[4,5-с] quinoline;

8-chloro-1-[cis-2-(difluoromethyl)tetrahydro-2H-pyran-4-yl]-2-[(5-methyl-1,2-oxazol-3-yl)methyl]-1H-imidazo[ 4,5-c]quinoline, ENT 1 (enantiomer 1);

8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-[(5-methyl-1,2-oxazol-3-yl)methyl]-1H-imidazo[4.5 -c]quinoline, ENT 1 (enantiomer 1);

1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-2-(1,3-thiazol-2-ylmethyl)-1H-imidazo[4,5-c]quinolin-8- carbonitrile;

8-chloro-1-[(3R)-1-methylpyrrolidin-3-yl]-2-[(4-methyl-1H-1,2,3-triazol-1-yl)methyl]-1H-imidazo[4 ,5-c]quinoline;

1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-2-(1,2,3-thiadiazol-4-ylmethyl)-8-(trifluoromethyl)-1H-imidazo[4, 5-c]quinoline;

8-fluoro-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-2-(1,3-thiazol-2-ylmethyl)-1H-imidazo[4,5-c] quinoline;

2-(1,3-benzoxazol-2-ylmethyl)-1-[cis-3-fluorocyclopentyl]-1H-imidazo[4,5-c]quinoline-8-carbonitrile;

1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-2-(1H-1,2,4-triazol-1-ylmethyl)-8-(trifluoromethyl)-1H-imidazo[ 4,5-c]quinoline;

8-chloro-2-[(5-methylpyrazin-2-yl)methyl]-1-[(3R)-1-methylpyrrolidin-3-yl]-1H-imidazo[4,5-c]quinoline;

1-[cis-3-fluorocyclopentyl]-2-[(5-methyl-1,2-oxazol-3-yl)methyl]-1H-imidazo[4,5-c]quinoline-8-carbonitrile;

1-[(2R,4R-2-methyltetrahydro-2H-pyran-4-yl]-2-(1,3-thiazol-4-ylmethyl)-8-(trifluoromethyl)-1H-imidazo[4,5-c ]quinoline;

2-[(5-methyl-1,3,4-oxadiazol-2-yl)methyl]-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-8-(trifluoromethyl) -1H-imidazo[4,5-c]quinoline;

8-chloro-1-(2,2-dimethyltetrahydro-2H-pyran-4-yl)-2-(1H-1,2,4-triazol-1-ylmethyl)-1H-imidazo[4,5-c] quinoline;

8-chloro-1-(2,2-difluoropropyl)-2-[(4-methoxy-1H-pyrazol-1-yl)methyl]-1H-imidazo[4,5-c]quinoline;

8-fluoro-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-2-{[5-(trifluoromethyl)pyrazin-2-yl]methyl}-1H-imidazo[4, 5-c]quinoline;

1-[(2R,4R-2-methyltetrahydro-2H-pyran-4-yl]-2-[(5-methyl-1,3,4-thiadiazol-2-yl)methyl]-8-(trifluoromethyl)- 1H-imidazo[4,5-c]quinoline;

8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-[(5-methyl-1,2,4-oxadiazol-3-yl)methyl]-1H-imidazo[4 ,5-c]quinoline, ENT 1 (enantiomer 1);

2-[(6-methylpyrimidin-4-yl)methyl]-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-8-(trifluoromethyl)-1H-imidazo[4,5 -c]quinoline;

8-chloro-1-[cis-3-fluorocyclopentyl]-2-[(5-methylpyrazin-2-yl)methyl]-1H-imidazo[4,5-c]quinoline;

3-{8-chloro-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[4,5-c]quinolin-2-yl}-2-methylpropanenitrile, DIAST 2 (diastereomer 2);

8-fluoro-1-[cis-3-fluorocyclopentyl]-2-(1,2,3-thiadiazol-4-ylmethyl)-1H-imidazo[4,5-c]quinoline, ENT 2 (enantiomer 2);

3-{8-chloro-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[4,5-c]quinolin-2-yl}propanenitrile;

1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-2-[(5-methyl-2H-tetrazol-2-yl)methyl]-8-(trifluoromethyl)-1H-imidazo [4,5-c]quinoline;

8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(4-methyl-1H-1,2,3-triazol-1-yl)methyl]-1H-imidazo [4,5-c]quinoline;

8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-(1H-tetrazol-1-ylmethyl)-1H-imidazo[4,5-c]quinoline;

8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(1-methyl-1H-1,2,4-triazol-3-yl)methyl]-1H-imidazo [4,5-c]quinoline;

1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-8-fluoro-2-[(2-methylimidazo[2,1-b][1,3,4]thiadiazol-6-yl)methyl ]-1H-imidazo[4,5-c]quinoline, ENT 1 (enantiomer 1);

8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-(1H-tetrazol-1-ylmethyl)-1H-imidazo[4,5-c]quinoline, ENT 2 (enantiomer 2);

8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(4-methyl-2H-1,2,3-triazol-2-yl)methyl]-1H-imidazo [4,5-c]quinoline, ENT 1 (enantiomer 1);

8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(5-methyl-1,2,4-oxadiazol-3-yl)methyl]-1H-imidazo[4 ,5-c]quinoline, ENT 1 (enantiomer 1);

8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(5-methyl-1,3,4-thiadiazol-2-yl)methyl]-1H-imidazo[4 ,5-c]quinoline, ENT 2 (enantiomer 2);

8-(difluoromethyl)-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-2-[(4-methyl-1H-1,2,3-triazol-1-yl) methyl]-1H-imidazo[4,5-c]quinoline;

8-(difluoromethyl)-2-[(5-methyl-1,2-oxazol-3-yl)methyl]-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H -imidazo[4,5-c]quinoline;

8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-[(5-methyl-1,3,4-thiadiazol-2-yl)methyl]-1H-imidazo[4 ,5-c]quinoline, ENT 1 (enantiomer 1);

8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-(1H-1,2,4-triazol-1-ylmethyl)-1H-imidazo[4,5-c] quinoline, ENT 1 (enantiomer 1);

8-chloro-2-[(4-cyclopropyl-1H-1,2,3-triazol-1-yl)methyl]-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-1H-imidazo [4,5-c]quinolone, ENT 1 (enantiomer 1); or

[5-({8-chloro-1-[(2R,4R-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[4,5-c]quinolin-2-yl}methyl)pyrazin- 2-yl]methanol

or a pharmaceutically acceptable salt thereof.

In another embodiment, the compound of the present invention is:

8-chloro-2-{[5-( ² H ₃ )methylpyrazin-2-yl]methyl}-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[ 4,5-c]quinoline;

8-chloro-2-{[5-( ² H ₃ )methylpyrazin-2-yl]( ² H ₂ )methyl}-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl] -1H-imidazo[4,5-c]quinoline;

8-chloro-2-{[5-( ² H ₂ )methylpyrazin-2-yl]methyl}-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[ 4,5-c]quinoline;

8-chloro-2-{[5-( ² H ₁ )methylpyrazin-2-yl]methyl}-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[ 4,5-c]quinoline;

[5-({8-chloro-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[4,5-c]quinolin-2-yl}methyl)pyrazine -2-yl]( ² H ₂ ) methanol; or

[5-({8-chloro-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[4,5-c]quinolin-2-yl}methyl)pyrazine -2-yl]( ² H ₁ ) methanol,

or a pharmaceutically acceptable salt thereof.

In another embodiment of the present invention, the compound of formula I contains R ¹ which is ethyl or

R ² is

R ³ is chlorine, cyano, difluoromethyl or trifluoromethyl, or a pharmaceutically acceptable salt thereof.

In another embodiment, the compound of formula I contains R ¹ which is

R ² is

R ³ is chlorine or cyano,

or a pharmaceutically acceptable salt thereof.

In another embodiment, the compound of the present invention of formula I contains R ¹ which is

R ² is

R ³ is chlorine,

or a pharmaceutically acceptable salt thereof.

In yet another embodiment, the compound of the present invention of formula I contains R ¹ which is

иR ² is

and

R ³ is chlorine,

or a pharmaceutically acceptable salt thereof.

In another embodiment, the present invention relates to a method of treating a disease or disorder selected from the group consisting of Crohn's disease, Parkinson's disease, Lewy body dementia, frontotemporal dementia, corticobasal dementia, progressive supranuclear palsy, leprosy, Alzheimer's disease, tauopathy and alpha -synucleinopathy, in a patient, comprising administering to a patient in need of such treatment a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

In yet another embodiment of the present invention, the treatment of the disease or disorder is selected from the group consisting of Crohn's disease, Parkinson's disease, Lewy body dementia, frontotemporal dementia, corticobasal dementia, progressive supranuclear palsy, leprosy, Alzheimer's disease, tauopathy, and alpha synucleinopathy.

In another embodiment, the treatment for the disease or disorder is selected from the group consisting of Lewy body dementia, frontotemporal dementia, corticobasal dementia, progressive supranuclear palsy, leprosy, inflammatory bowel disease, irritable bowel syndrome, Alzheimer's disease, tauopathy and alpha synucleinopathy, disease Parkinson's disease, Parkinson's disease with dementia, syndrome at risk of developing Parkinson's disease, Alzheimer's disease with Lewy bodies, combination of Parkinson's disease and Alzheimer's disease, multiple system atrophy, striatonigral degeneration, olivopontocerebellar atrophy, Shy-Drager syndrome, ulcerative colitis, juvenile parkinsonism, disease Steele-Richardson-Olszewski, Lytico-Bodig disease or Guam complex (ALS (amyotrophic lateral sclerosis)-parkinsonism-dementia), corticobasal ganglion degeneration, progressive pallidar atrophy, parkinsonism-dementia complex, striopallidar syndrome, hereditary juvenile dystonia-parkinsonism, autosomal dominant Lewy body disease, Huntington's disease, Wilson's disease, hereditary ceruloplasmin deficiency, Hallervorden-Spatz disease, olivopontocerebellar and spinocerebellar attacks with retinal degeneration, Machado-Joseph disease, familial amyotrophy-dementia-parkinsonism, disinhibition complex- dementia-parkinsonism-amyotrophy, Gerstmann-Streussler-Scheinker syndrome, familial progressive subcortical gliosis, Lubag syndrome (X-linked dystonia-parkinsonism), familial basal ganglia calcification, mitochondrial cytopathies with striatal necrosis, waxy lipofuscinosis, familial parkinsonism with peripheral neuropathy, parkinsonism -pyramidal syndrome, neuroacanthocytosis and hereditary hemochromatosis.

In another embodiment, the present invention provides a method of treating a disease or disorder selected from a neurological disease, most preferably Parkinson's disease (but also other neurological diseases such as migraine; epilepsy; Alzheimer's disease; Niemann-Pick type C; brain injury; stroke ; cerebrovascular disease; cognitive disorder; sleep disorder) or psychiatric disorder (such as anxiety; feigning disorder; impulse disorder; mood disorder; psychomotor disorder; psychotic disorder; drug dependence; eating disorder and psychiatric disorder in children), in a mammal, preferably a human, comprising administering to said mammal a therapeutically effective amount of a compound of Formula I or a pharmaceutically acceptable salt thereof. In addition, the compounds of formula I and their pharmaceutically acceptable salts can also be used in methods of treating other LRRK2-associated disorders such as Crohn's disease, leprosy, and certain cancers such as kidney, breast, lung, prostate, and blood cancers. .

The Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, (DSM-IV-TR) (2000, American Psychiatric Association, Washington) provides a diagnostic tool for identifying many of the disorders described here. One skilled in the art understands that there are alternative nomenclatures, nosological classifiers, and classification systems for the disorders described herein, including those described in the DMS-IV-TR, and that terminology and classification systems evolve in the course of scientific progress in medicine.

General synthesis schemes

Compounds of formula I can be obtained by the methods described below, together with methods of synthesis known in the field of organic chemistry, or through modifications and transformations obvious to experts in this field of technology. The starting materials used here are commercially available or can be prepared by standard methods known in the art [such as those disclosed in standard manuals such as Compendium of Organic Synthetic Methods, Vol. I-XII (published by Wiley-Interscience)]. Preferred methods include, but are not limited to, those described below.

During the following synthetic sequences, it may be necessary and/or desirable to protect sensitive or reactive groups on any molecules of interest. This can be achieved with conventional protecting groups such as those described in T.W. Greene, Protective Groups in Organic Chemistry, John Wiley & Sons, 1981; T.W. Greene and P.G.M. Wuts, Protective Groups in Organic Chemistry, John Wiley & Sons, 1991; and T.W. Greene and P.G.M. Wuts, Protective Groups in Organic Chemistry, John Wiley & Sons, 1999, which are incorporated here by reference.

Compounds of formula I or their pharmaceutically acceptable salts can be obtained in accordance with the schemes of the reactions, which are discussed below. Unless otherwise stated, substituents in the schemes are as defined above. The isolation and purification of the products is carried out by standard techniques which are known to the skilled chemist.

One skilled in the art will appreciate that in many cases the compounds in Reaction Schemes 1-9 can be obtained as a mixture of diastereomers and/or enantiomers; they can be separated at different stages of synthetic schemes using standard techniques and a combination of techniques such as, for example, crystallization, normal phase chromatography, reversed phase chromatography and chiral chromatography, to obtain individual enantiomers of the invention.

One skilled in the art will appreciate that the various symbols, superscripts, and subscripts used in the diagrams, methods, and examples are used for convenience of presentation and/or to display the order in which they are presented in the diagrams, and are not intended to to be sure to match the symbols, superscripts, and subscripts in the appended claims. The schemes are an illustrative representation of methods useful in the synthesis of the compounds of the present invention. They do not limit the scope of the present invention in any way.

Interactions to obtain compounds of the invention can be carried out in suitable solvents, which can be easily selected by a person skilled in the field of organic synthesis. Suitable solvents may be substantially unreacted with the starting materials (reagents), intermediates or products at temperatures at which the reactions are carried out, for example at temperatures that may range from the freezing point of the solvent to the boiling point of the solvent. A given reaction may be carried out in a single solvent or a mixture of more than one solvent. Depending on the particular reaction step, suitable solvents may be selected by one skilled in the art for the particular reaction step.

Interactions can be controlled according to any suitable method known in the art. For example, product formation can be monitored by means of spectroscopy, such as nuclear magnetic resonance spectroscopy (for example, ¹ H or ¹³ C), infrared spectroscopy, spectrophotometry (for example, in the UV-visible regions), mass spectrometry, or by chromatographic methods such as as high performance liquid chromatography (HPLC) or thin layer chromatography (TLC).

Compounds of formula I and their intermediates can be prepared according to the following reaction schemes and their accompanying discussion. Unless otherwise noted, R ¹ , R ² and R ³ in the reaction schemes and subsequent discussions are as defined above. In General, the compounds of the present invention can be obtained by methods, including methods similar to those known in the field of chemistry, in particular in light of the description contained here. Several methods for preparing the compounds of this invention and their intermediates are provided as further aspects of the invention and are illustrated by the following reaction schemes. Other methods may be described in the experimental section. The schemes and examples provided here (including the corresponding description) are for illustration only and are not intended to limit the scope of the present invention.

Reaction Scheme 1

Reaction Scheme 1 shows the preparation of compounds of formula (I). Referring to scheme 1, the compounds of formula 1.1 and 1.2 are either commercially available or can be obtained by the methods described here, or other methods well known to those skilled in the art. In a compound of formula 1.1, the group denoted LG is a suitable leaving group, such as a halide (eg chlorine or bromine) or triflate, which is suitable to be subjected to nucleophilic substitution when reacted with an amine of formula 1.2. In the amine of Formula 1.2, the group designated PG is a suitable amine protecting group, such as an acid-labile protecting group selected from 2,4-dimethoxybenzyl (DMB), 4-methoxybenzyl (PMB), and tert-butoxycarbonyl (Boc). The compounds of formulas 1.1 and 1.2 may be reacted, for example, in the presence of a suitable base such as N,N-diisopropylethylamine (Hunig's base) or triethylamine, in a suitable solvent such as acetonitrile or N,N-dimethylformamide (DMF) to give the compound formulas 1.3. The interaction is usually carried out at an elevated temperature, such as from 50 to 100°C for a period of 1 to 48 hours. Removal of a protecting group, such as an acid-labile protecting group (PG), from a compound of formula 1.3 can generally be accomplished by treating a compound of formula 1.3 with an appropriate acid, such as acetic acid, trifluoroacetic acid, or hydrochloric acid, to give a compound of formula 1.4. It should also be understood that, in some cases, a compound of formula 1.1 may be reacted with an unprotected amine of formula R ² -NH ₂ to give the compound of formula 1.4 directly. Reduction of the nitro group in a compound of formula 1.4 using conditions suitable for the functional group present gives a compound of formula 1.5. For example, the nitro group in a compound of formula 1.4 can be reduced to the corresponding amine of formula 1.5 by treating a compound of formula 1.4 with zinc dust and ammonium hydroxide in methanol, or alternatively by hydrogenating a compound of formula 1.4 using a suitable catalyst such as platinum(IV) oxide in a suitable solvent such as methanol, acetonitrile, or a mixture thereof. Coupling of a diamine of formula 1.5 with a carboxylic acid of formula 1.6 then gives the desired compound of formula I, also designated 1.7. The coupling reaction with a diamine of formula 1.5 and a carboxylic acid of formula 1.6 can be carried out in an appropriate solvent such as N,N-dimethylformamide or N-propyl acetate in the presence of a suitable base such as N,N-diisopropylethylamine and a coupling agent such as 2 ,4,6-tripropyl-1,3,5,2,4,6-trioxatrifosfinan 2,4,6-trioxide or 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI). The coupling reaction is often carried out by heating from 60°C to 110°C.

Reaction Scheme 2

Reaction Scheme 2 shows the preparation of a compound of formula 1.7' which is a compound of formula I in which R ² is shown to be a chiral 2-methyltetrahydropyran-4-yl moiety. Using a published procedure, a Prins reaction of compound 2.1 with a compound of formula 2.2 produces a pyran of formula 2.3. Chiral resolution to obtain separate enantiomers using an enzymatic method gives the compound of formula 2.5 after hydrolysis of the separated ester of formula 2.4. Oxidation of a compound of formula 2.5 yields a ketone of formula 2.6 which is reacted with a compound of formula 2.7 using reductive amination to give a protected amine of formula 2.8. The protected amine of formula 2.8 may be reacted with a compound of formula 1.1 in a manner analogous to that previously described in Scheme 1 to give a compound of formula 1.3'. The compounds of formulas 1.4', 1.5' and 1.7' can then be obtained by a method similar to the methods described in scheme 1 for compounds of formulas 1.4, 1.5 and 1.7, respectively.

Reaction Scheme 3

Reaction Scheme 3 shows the preparation of a compound of formula 3.13 which is a compound of formula I wherein R ² is shown to be a chiral 2-cyanomethyltetrahydropyran-4-yl moiety. Using the published procedure, the Prins reaction of compound 3.1 with but-3-en-1-ol gave pyran of formula 3.2. Oxidation of the compound of formula 3.2 gave the ketone of formula 3.3, which was reacted with dimethoxybenzylamine using reductive amination to give the protected amine of formula 3.4. The protected amine of formula 3.4 may be reacted with a compound of formula 1.1 in a manner analogous to that previously described in Scheme 1 to give a compound of formula 3.5. Removal of the protecting group under acidic conditions gave the compound of formula 3.6. The nitro group of a compound of formula 3.6 is reduced by catalytic hydrogenation or by treatment with a metal such as zinc or iron to give a diamine of formula 3.7. Acylation of a compound of formula 3.7 with an acid of formula 3.8 under various coupling conditions known to those skilled in the art provides a compound of formula 3.9. The amide of formula 3.9 can be dehydrated under thermal conditions to give a compound of formula 3.10. Deprotection of the compound of formula 3.10 with a Lewis acid such as BCl ₃ , TMSI, AlCl ₃ or by palladium catalyzed hydrogenation gives the alcohol of formula 3.11. An alcohol of formula 3.11 can be converted to an activated leaving group such as a sulfonate, such as but not limited to a mesylate of formula 3.12. The compounds of formulas 3.13 can then be obtained by nucleophilic substitution of the mesylate with a cyanide anion.

Reaction Scheme 4

Reaction Scheme 4 shows the preparation of a compound of formula 4.8 which is a compound of formula I wherein R ² is shown to be a chiral 2-methyltetrahydropyran-4-yl moiety and R ³ is cyano. The interaction begins with a known acid of formula 4.1, which is subjected to interaction with N-hydroxy-2-nitroethanamine obtained in situ to obtain the compound of formula 4.2. Nitroamine 4.2 was treated with a carboxylic acid activating agent followed by condensation to give the quinolone of formula 4.3. The phenol of formula 4.3 can be converted to the activated chloride of formula 4.4 with phosphorus oxychloride or thionyl chloride. The chloride of formula 4.4 can be subjected to nucleophilic substitution with an appropriate amine, such as a compound of formula 2.8, to give a compound of formula 4.5. A compound of formula 4.5 can be deprotected to give a compound of formula 4.6 which in turn is reduced to give a diamine of formula 4.7. Compounds of formula 4.8 can be prepared from a compound of formula 4.7 by condensation with the appropriate acid R ¹ CO ₂ H using a method similar to that described previously.

Reaction Scheme 5

Reaction Scheme 5 shows the preparation of a compound of formula 5.6 which is a compound of formula I wherein R ² is shown to be a chiral 2-methyltetrahydropyran-4-yl moiety and R ³ is a difluoromethyl group. The compound of formula 5.1 is treated with 2,2-difluoro-1-phenylethan-1-one and a suitable palladium complex such as cataCXium A Pd G2 and a base such as tripotassium phosphate n-hydrate in an inert solvent such as toluene to give compounds of formula 5.2. The benzoyl group of a compound of formula 5.2 can be removed with a base such as sodium hydroxide or potassium hydroxide in water or other similar conditions. Alternatively, the benzoyl is removed in an alcoholic solvent with sodium methoxide. The protecting group of a compound of formula 5.3 (such as a DMB group) can be removed as previously described and the nitro group of a compound of formula 5.4 can be reduced to give a diamine of formula 5.5. Compounds of formula 5.6 can be prepared from a compound of formula 5.5 using a method similar to that described previously, by condensing a compound of formula 5.5 with the appropriate acid R ¹ CO ₂ H.

Reaction Scheme 6

Reaction Scheme 6 shows the preparation of a compound of formula 6.9 which is a compound of formula I wherein R ² is shown to be a chiral 4,4-difluoro-1-methylpyrrolidin-3-yl moiety and R ³ is cyano. This amine is prepared by the method described in U.S. Published Application 20150141402. This kind of compound can be prepared as in the examples above by forming a chloride of formula 6.3 by reacting a compound of formula 6.2 with phosphorus oxychloride or thionyl chloride in a suitable inert solvent. The chloride is treated with an amine of formula 6.4 in the presence of a suitable base such as Hunig's base (N,N-diisopropylethylamine) or triethylamine to give a compound of formula 6.5. The protecting group is removed by treating the compound of formula 6.5 with an acid such as trifluoroacetic acid or hydrochloric acid. The secondary amine of formula 6.6 can be methylated by standard reductive amination using formaldehyde and a reducing agent such as sodium triacetoxyborohydride or sodium cyanoborohydride. The nitro group of a compound of formula 6.7 may be reduced by hydrogenation over a platinum catalyst, or alternatively the nitro group may be reduced with a suitable metal such as iron or zinc. The claimed compounds of formula 6.9 can be obtained from the compound of formula 6.8 by condensation with the appropriate acid R ¹ CO ₂ H under the conditions described previously.

Reaction Scheme 7

Reaction Scheme 7 shows the preparation of a compound of formula 7.5 which is a compound of formula I in which R ² is shown to be a chiral 3,3-difluorotetrahydro-2H-pyran-4-amine moiety. The chloride of formula 7.1 is treated with an amine of formula 7.2 in the presence of a suitable base such as Hunig's base or triethylamine to give a compound of formula 7.3. The nitro group of a compound of formula 7.3 may be reduced by hydrogenation over a platinum catalyst, or alternatively the nitro group may be reduced with a suitable metal such as iron or zinc. The compounds of formula 7.5 can then be obtained from the compound of formula 7.4 by condensation with the appropriate acid R ¹ CO ₂ H under the conditions described previously.

Reaction Scheme 8

Reaction Scheme 8 shows the preparation of a compound of formula 8.5 which is a compound of formula I wherein R ² is shown to be a chiral (R)-1-methylpyrrolidin-3-amine moiety and R ³ is cyano. The chloride is treated with a chiral amine of formula 8.2 in the presence of a suitable base such as Hunig's base or triethylamine to give a compound of formula 8.3. The nitro group of a compound of formula 8.3 may be reduced by hydrogenation over a platinum catalyst, or alternatively the nitro group may be reduced with a suitable metal such as iron or zinc. Compounds of formula 8.5 may be prepared from a compound of formula 8.4 by condensation with the appropriate acid R ¹ CO ₂ H under the conditions previously described.

Reaction Scheme 9

Reaction Scheme 9 shows the preparation of a compound of formula 9.8 which is a compound of formula I wherein R ² is shown to be a chiral 2-methyltetrahydropyran-4-yl moiety and R ³ is trifluoromethyl. The chloride of formula 9.3 is treated with an amine of formula 2.8 in the presence of a suitable base such as Hunig's base or triethylamine to give a compound of formula 9.5. Removal of the protecting group under acidic conditions gives the compound of formula 9.6. The nitro group of a compound of formula 9.6 may be reduced by hydrogenation over a platinum catalyst, or alternatively the nitro group may be reduced with a suitable metal such as iron or zinc. The claimed compounds of formula 9.8 can be obtained from the compound of formula 9.7 by condensation with the appropriate acid R ¹ CO ₂ H under the conditions described previously.

The general methods described in Schemes 1 to 9 should not be construed as limiting. The person skilled in the art should understand that changes in the order of certain stages and reaction conditions can be made to obtain the compounds of formula I. The choice of which approach is best used can be made by one skilled in the art of organic synthesis. More specific examples of applicable methods for the preparation of compounds of formula I are provided below in the Examples section and furthermore, these methods are also not to be construed as limiting by those skilled in the art.

Experimental methods

The synthesis of various compounds of the present invention is illustrated below. Additional compounds within the scope of this invention can be obtained using the methods illustrated in these examples, either alone or in combination with methods generally known in the art.

молекулярными ситами, до достижения следующих стандартов контроля качества (QC) для воды: а) <100 м.д. для дихлорметана, толуола, N,N-диметилформамида и тетрагидрофурана; б) <180 м.д. для метанола, этанола, 1,4-диоксана и диизопропиламина. Для очень чувствительных реакций растворители дополнительно обрабатывали металлическим натрием, гидридом кальция или молекулярными ситами и дистиллировали непосредственно перед применением. Продукты в основном сушили под вакуумом перед проведением дальнейших взаимодействий или биологического тестирования. Данные масс-спектрометрии регистрировали на оборудовании для жидкостной хроматографии/масс-спектрометрии (LCMS), химической ионизации при атмосферном давлении (APCI) или газовой хроматографии/масс-спектрометрии (GCMS). Данные химических сдвигов для ядерного магнитного резонанса (NMR) выражены в миллионных долях (м.д., δ) относительно остаточных пиков использованных дейтерированных растворителей. В некоторых примерах хиральные разделения проводили, чтобы разделить энантиомеры или диастереомеры некоторых соединений по изобретению (в некоторых примерах раздельные энантиомеры обозначены как ENT 1 и ENT 2 в соответствии с порядком их элюирования, и раздельные диастереомеры обозначены как DIAST 1 и DIAST 2 в соответствии с порядком их элюирования). В некоторых примерах оптическое вращение энантиомера измеряли при использовании поляриметра. В соответствии с данными его наблюдаемого вращения (или данными его удельного вращения) энантиомер с вращением по часовой стрелке обозначали как (+)-энантиомер, а энантиомер с вращением против часовой стрелки обзначали как (-)-энантиомер. На рацемические соединения указывает наличие символов «(+/-)» перед структурой; в этих случаях указанная стереохимия представляет относительную (нежели абсолютную) конфигурацию заместителей в соединении.The experiments were mainly carried out in an inert gas atmosphere (nitrogen or argon), especially in cases where oxygen or humidity sensitive reagents or intermediates were used. Commercially available solvents and reagents were generally used without further purification. Anhydrous solvents were used where appropriate, mainly AcroSeal® products from Acros Organics, Aldrich Sure/Seal™ from Sigma-Aldrich, or DriSolv® products from EMD Chemicals. In other cases, commercially available solvents were passed through columns packed with 4

molecular sieves until the following quality control (QC) standards for water are achieved: a) <100 ppm. for dichloromethane, toluene, N,N-dimethylformamide and tetrahydrofuran; b) <180 ppm for methanol, ethanol, 1,4-dioxane and diisopropylamine. For very sensitive reactions, the solvents were further treated with sodium metal, calcium hydride, or molecular sieves and distilled immediately before use. The products were generally dried under vacuum prior to further interactions or biological testing. Mass spectrometry data were recorded on liquid chromatography/mass spectrometry (LCMS), atmospheric pressure chemical ionization (APCI), or gas chromatography/mass spectrometry (GCMS) equipment. Chemical shift data for nuclear magnetic resonance (NMR) are expressed in parts per million (ppm, δ) relative to the residual peaks of the deuterated solvents used. In some examples, chiral separations were performed to separate the enantiomers or diastereomers of some compounds of the invention (in some examples, the separate enantiomers are designated ENT 1 and ENT 2 according to their elution order, and the separate diastereomers are designated DIAST 1 and DIAST 2 according to the order their elution). In some examples, the optical rotation of an enantiomer was measured using a polarimeter. According to its observed rotation data (or its specific rotation data), the clockwise rotation enantiomer was designated as the (+)-enantiomer, and the counter-clockwise rotation enantiomer was designated as the (-)-enantiomer. Racemic compounds are indicated by the presence of the symbols "(+/-)" before the structure; in these cases, said stereochemistry represents the relative (rather than absolute) configuration of the substituents on the compound.

Reactions carried out with detectable intermediates were typically monitored by LCMS and allowed to proceed until complete conversion before adding further reagents. For standard synthesis procedures in other examples or methods, the reaction conditions (temperature and reaction time) can be changed. In general, after the interactions, thin layer chromatography or mass spectrometry is carried out and, if necessary, subjected to processing. Purification methods may vary depending on the experiments: in general, the solvents and solvent ratios used for the eluents/gradients were chosen to obtain appropriate R _f values or retention times. All of the starting materials in these preparations and examples are either commercially available or can be obtained by methods known in the art or as described here.

Interactions were carried out in air or in an atmosphere of inert gas (nitrogen or argon) using reagents or intermediates that are sensitive to oxygen or humidity. If necessary, the equipment was dried under dynamic vacuum using a hot air gun and anhydrous solvents (Sure-SealTM products from Aldrich Chemical Company, Mil-waukee, Wisconsin or DriSolvTM products from EMD Chemicals, Gibbstown, NJ) were used. Commercially available solvents and reagents were used without further purification. When indicated, the interactions were carried out under heating by microwave irradiation using Biotage Initiator or Personal Chemistry Emrys Optimizer microwave reactors or the like. The progress of the reaction was monitored using thin layer chromatography (TLC), liquid chromatography-mass spectrometry (LCMS) and high performance liquid chromatography (HPLC) assays. TLC was performed on pre-coated silica gel plates with a fluorescent indicator (excitation wavelength 254 nm) and visualized under UV light and/or with dyes such as I ₂ , KMnO ₄ , CoCl ₂ , phosphomolybdic acid and/or ceric ammonium molybdate. LCMS data were recorded on an Agilent 1100 Series detector with a Leap Technologies autosampler, Gemini C18 columns, MeCN/water gradients, and either TFA, formic acid, or ammonium hydroxide modifiers, or similar equipment. The eluate was analyzed using a Waters ZQ spectrometer, scanning in the detection mode of both positively and negatively charged ions from 100 to 1200 Da. Other similar devices were also used. HPLC data were recorded on an Agilent 1100 Series detector using Gemini or XBridge C18 columns, with MeCN/water gradients and TFA or ammonium hydroxide modifiers, and similar equipment. Purification was performed by medium pressure liquid chromatography (MPLC) using Isco CombiFlash Companion, AnaLogix IntelliFlash 280, Biotage SP1 or Biotage Isolera One instruments and Isco RediSep or Biotage Snap prepackaged silica cartridges and the like. Chiral purification was performed by chiral supercritical fluid chromatography (SFC) using Berger or Thar instruments and similar instruments; columns Chi-ralPAK-AD, -AS, -IC, Chiralcel-OD or -OJ; and mixtures of CO ₂ with MeOH, EtOH, iPrOH or MeCN, alone or modified using TFA or iPrNH ₂ . UV detection was used for triggered collection of fractions.

Mass spectrometry data were obtained from LCMS analyses. Mass spectrometry (MS) was performed using atmospheric pressure chemical ionization (APCI), electrospray ionization (ESI), electron impact ionization (EI), or electron scattering ionization (ES) sources. Proton nuclear magnetic resonance (1H NMR) chemical shifts are given in ppm in the direction of the lower magnetic field away from tetramethylsilane and were recorded on Varian spectrometers at 300, 400, 500, or 600 MHz. Chemical shifts are expressed in parts per million (ppm, δ) relative to the residual peaks of the deuterated solvents used. Peak shapes are described as follows: s, singlet; d, doublet; t, triplet; q, quartet; quin, quintet; m, multiplet; br s, broadened singlet; app, apparent. Analytical SFC data were obtained on a Berger analytical instrument as described above. Optical rotation data were obtained on a PerkinElmer Model 343 polarimeter using a 1 dm cell. Silica gel chromatography was predominantly performed using Biotage or ISCO medium pressure systems using prepackaged columns from various commercial vendors including Biotage and ISCO.

Unless otherwise noted, chemical interactions were carried out at room temperature (approximately 23 degrees Celsius).

The compounds and intermediates described below were named using the naming conventions provided by the ACD/ChemSketch 2012, File Version C10H41, Build 69045 software package (Advanced Chemistry Development, Inc., Toronto, Ontario, Canada). The naming conventions provided by the ACD/ChemSketch 2012 software package are well known to those skilled in the art and it is assumed that the naming conventions provided by the ACD/ChemSketch 2012 software package generally follow the IUPAC (International Union of Pure and Applied Chemistry) recommendations for nomenclature organic compounds and CAS identifier rules (Chemical Abstract Service).

In the experimental sections that follow, the following abbreviations may be used. ACN means acetonitrile; Ac ₂ O is acetic anhydride; wide means broad; °C means degrees Celsius; CDCl ₃ means deuterochloroform; CD ₃ OD is deuteromethanol; CH ₃ NO ₂ means nitromethane; d means doublet; DCM means dichloromethane; DEA means diethylamine; DIAST means diastereomer; DIEA means N,N-diisopropylethylamine; DMB means dimethoxybenzyl; DMSO means dimethyl sulfoxide; EDCI means 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride; ENT means enantiomer; EtOAc means ethyl acetate; EtOH means ethanol; ES means electrospray; FA stands for formic acid; g means gram; h means hour; HCl means hydrochloric acid; H ₂ means hydrogen; H ₂ O means water; HPLC stands for High Performance Liquid Chromatography; Hz means Hertz; K ₂ CO ₃ means potassium carbonate; l means liter; LC means liquid chromatography; LCMS stands for liquid chromatography-mass spectrometry; m means multiplet; M means molar; MeOH means methanol; MgSO ₄ means magnesium sulfate; MHz means megahertz; min means minute; ml means milliliter, mM means millimoles; μl means microliter; μM means micromole; MS means mass spectrometry; MsCl is methanesulfonyl chloride; MTBE means methyl tert-butyl ether; NADPH means nicotinamide adenine dinucleotide phosphate; N ₂ means nitrogen; NEt ₃ means triethylamine; NaHCO ₃ means sodium bicarbonate; Na ₂ SO ₄ means sodium sulfate; NH ₄ Cl means ammonium chloride; NH ₄ HCO ₃ means ammonium bicarbonate; NH ₄ OH means ammonium hydroxide; NMR stands for nuclear magnetic resonance; PE means petroleum ether; psi means pounds per square inch; Pt/C means platinum on carbon; RT means retention time or room temperature depending on the context; s means singlet; SFC stands for supercritical fluid chromatography; t means triplet; TFA means trifluoroacetic acid; THF means tetrahydrofuran; TLC means thin layer chromatography; and T3P is propylphosphinic acid anhydride.

Preparation of compound P1

(2R,4R)-N-(2,4-Dimethoxybenzyl)-2-methyltetrahydro-2H-pyran-4-amine (Р1)

Stage 1. Synthesis of cis-2-methyltetrahydro-2H-pyran-4-ol (C1).

But-3-en-1-ol (39.0 ml; 453 mmol) and acetaldehyde (25.5 ml; 454 mmol) were combined in aqueous sulfuric acid (20% (w/w); 565 g) and stirred at 80°C for 5 days. The reaction mixture was cooled to room temperature and extracted with diethyl ether and then with dichloromethane; the combined organic layers were dried over magnesium sulfate, filtered and concentrated in vacuo. Chromatography on silica gel (gradient: 0% to 25% ethyl acetate in heptane) provided the product as a colorless oil. Yield: 11.2 g; 96.4 mmol; 21%. ¹ H NMR (400 MHz, CDCl ₃ ) δ 3.99 (ddd, J=11.8, 4.9, 1.7 Hz, 1H), 3.71-3.80 (m, 1H), 3.35-3.46 (m, 2H) , 1.82-1.98 (m, 3H), 1.48 (dddd, J=12.5, 12.4, 11.1, 4.9 Hz, 1H), 1.21 (d, J=6.2 Hz, 3H), 1.14-1.24 (m, 1H).

Stage 2. Synthesis of (2R,4R)-2-methyltetrahydro-2H-pyran-4-ylbutanoate (C2).

Ethenyl butanoate (78.6 ml; 620 mmol) and Novozyme 435 (immobilized Candida antarctica yeast lipase B; 25 g) were added to a solution of Compound C1 (150 g; 1.29 mol) in tetrahydrofuran (1.3 L). The reaction mixture was stirred at room temperature for 2 hours, after which it was filtered through a pad of diatomaceous earth, which was then washed twice with dichloromethane. The combined filtrates were concentrated in vacuo and purified by silica gel chromatography (gradient: 0% to 10% ethyl acetate in heptane) to give the product as an oil. Yield: 51.5 g; 276 mmol; 45%. The absolute configuration of compound C2 and the following intermediates was confirmed by X-ray diffraction analysis performed on compound C32 (see preparation of compound P10). ¹ H NMR (400 MHz, CDCl ₃ ) δ 4.82-4.92 (m, 1H), 3.99 (ddd, J=11.9, 4.9, 1.7 Hz, 1H), 3.42-3.52 (m, 2H) , 2.25 (t, J=7.4 Hz, 2H), 1.92-2.00 (m, 1H), 1.84-1.91 (m, 1H), 1.52-1.69 (m, 3H), 1.28 (ddd, J=12, 11, 11 Hz, 1H), 1.20 (d, J=6.2 Hz, 3H), 0.94 (t, J=7.4 Hz, 3H).

Stage 3. Synthesis of (2R,4R)-2-methyltetrahydro-2H-pyran-4-ol (C3).

A solution of compound C2 (51.5 g; 276 mmol) in methanol and tetrahydrofuran (1:1 mixture; 700 ml) was treated with a solution of lithium hydroxide (19.9 g; 831 mmol) in water (120 ml) and the reaction mixture was stirred in overnight at room temperature. After removal of organic solvents by concentration under reduced pressure, the aqueous residue was extracted 4 times with dichloromethane; the combined organic layers were dried over magnesium sulfate, filtered and concentrated in vacuo to give the product as a colorless oil. Yield: 27.3 g; 235 mmol; 85%. ¹ H NMR (400 MHz, CDCl ₃ ) δ 3.99 (ddd, J=11.8, 4.8, 1.7 Hz, 1H), 3.71-3.80 (m, 1H), 3.35-3.47 (m, 2H) , 1.82-1.98 (m, 3H), 1.48 (dddd, J=12.5, 12.4, 11.1, 4.8 Hz, 1H), 1.21 (d, J=6.2 Hz, 3H), 1.14- 1.24(m, 1H).

Stage 4. Synthesis of (2R)-2-methyltetrahydro-4H-pyran-4-one (C4).

A solution of compound C3 (27.3 g; 235 mmol) in acetone (980 ml) was cooled in an ice bath and treated dropwise with Jones' reagent (2.5 M; 103 ml; 258 mmol). The reaction mixture was stirred for 10 minutes at 0°C, then warmed to room temperature, stirred for another 30 minutes, and cooled to 0°C. 2-Propanol (18 ml; 240 mmol) was added and stirring was continued for 30 minutes. After the mixture was concentrated under vacuum, the residue was partitioned between water and dichloromethane; the aqueous layer was extracted 3 times with dichloromethane, and the combined organic layers were dried over magnesium sulfate, filtered, and concentrated under reduced pressure to give the product as a light yellow oil. Yield: 23 g; 200 mmol; 85%. ¹ H NMR (400 MHz, CDCl ₃ ) δ 4.25 (ddd, J=11.5, 7.4, 1.3 Hz, 1H), 3.70 (dqd, J=12.2, 6.1, 2.7 Hz, 1H), 3.64 (ddd, J=12.2, 11.6, 2.8 Hz, 1H), 2.55 (dddd, J=14.6, 12.4, 7.4, 1.0 Hz, 1H), 2.37 (ddd, J=14.4, 2.3, 2.3 Hz, 1H), 2.21-2.31 (m, 2H), 1.29 (d, J=6.2 Hz, 3H).

Step 5. Synthesis of (2R,4R)-N-(2,4-dimethoxybenzyl)-2-methyltetrahydro-2H-pyran-4-amine (P1).

1-(2,4-Dimethoxyphenyl)methanamine (20.3 ml; 135 mmol) was added to a solution of compound C4 (10.3 g; 90.2 mmol) in methanol (200 ml) and the reaction mixture was stirred for 1 hour at room temperature. Then it was cooled to -78°C; a solution of lithium borohydride (2 M in tetrahydrofuran; 45.1 ml; 90.2 mmol) was added dropwise and stirring was continued at -78° C. for 2 hours. After slowly warming to room temperature overnight, the reaction mixture was quenched by careful addition of saturated aqueous sodium bicarbonate. Added ethyl acetate (250 ml) and enough water to dissolve the precipitate, and the aqueous layer was extracted with ethyl acetate; the combined organic layers were dried over magnesium sulfate, filtered and concentrated in vacuo. Chromatography on silica gel (gradient: 0% to 5% methanol in dichloromethane) gave the product as a colorless oil (10.4 g). Similar purification of mixed fractions gave additional product (3.7 g). Total yield: 14.1 g; 53.1 mmol; 59%. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.13 (d, J=8.0 Hz, 1H), 6.42-6.47 (m, 2H), 3.99 (ddd, J=11.6, 4.6, 1, 5 Hz, 1H), 3.82 (s, 3H), 3.80 (s, 3H), 3.76 (s, 2H), 3.36-3.45 (m, 2H), 2.63-2.73 (m, 1H), 1.85-1.92 (m , 1H), 1.78-1.85 (m, 1H), 1.38 (dddd, J=13, 12, 11, 4.7 Hz, 1H), 1.20 (d, J=6.2 Hz, 3H), 1.10 (ddd , J=11, 11, 11 Hz, 1H).

Preparation of compound P2

cis-2-[(Benzyloxy)methyl]-N-(2,4-dimethoxybenzyl)tetrahydro-2H-pyran-4-amine (Р2)

Stage 1. Synthesis of 2-[(benzyloxy)methyl]tetrahydro-2H-pyran-4-ol (C5).

A solution of (benzyloxy)acetaldehyde (25.0 g; 166 mmol) and but-3-en-1-ol (12.0 g; 166 mmol) in dichloromethane (550 ml) was added dropwise to a solution of trifluoroacetic acid (57 g; 500 mmol) in dichloromethane (500 ml) at 0°C. The reaction mixture was stirred at room temperature (20°C) for 18 hours, after which it was concentrated under vacuum. After the residue was dissolved in methanol (450 ml), it was treated with potassium carbonate (80 g; 580 mmol) and the reaction mixture was stirred for 5 hours at 20°C. The reaction mixture was added after a similar reaction using (benzyloxy)acetaldehyde (20.0 g; 133 mmol) and the combined mixtures were filtered. The filtrate was concentrated under reduced pressure and partitioned between water (500 ml) and ethyl acetate (200 ml). The aqueous layer was then extracted with ethyl acetate (2×150 ml) and the combined organic layers were concentrated in vacuo. Chromatography on silica gel (gradient: 20% to 25% ethyl acetate in petroleum ether) gave the product as a yellow oil. Based on the study of the ¹ H NMR spectrum, this substance was assumed to be a mixture of cis and trans isomers. Total yield: 42.9 g; 193 mmol; 64%. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.39-7.26 (m, 5H), 4.64-4.53 (m, 2H), [4.29-4.25 (m), 4.11-3.76 (m), and 3.59-3.40 (m ), total 6H], [1.96-1.83 (m), 1.71-1.48 (m), and 1.36-1.24 (m), total 4H, putative; partially covered by a peak of water].

Stage 2. Synthesis of 2-[(benzyloxy)methyl]tetrahydro-4H-pyran-4-one (C6).

Pyridinium chlorochromate (48 g; 220 mmol) was added to a solution of compound C5 (22.9 g; 103 mmol) in dichloromethane (350 ml) and the reaction mixture was stirred at room temperature (20° C.) for 18 hours. It was then combined with a similar reaction mixture prepared using compound C5 (20 g; 90 mmol) and the mixture was filtered, then concentrated in vacuo. The residue was purified by silica gel chromatography (eluent: 20% ethyl acetate in petroleum ether) to give the product as a colorless oil. Total yield: 36.2 g; 164 mmol; 85%. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.40-7.27 (m, 5H), 4.65-4.58 (m, 2H), 4.36 (ddd, J=11.5, 7.5, 1.5 Hz, 1H) , 3.85 (dddd, J=11.5, 4.3 Hz, 1H), 3.72 (ddd, J=12.3, 11.5, 2.8 Hz, 1H), 3.58 (dd, half ABX pattern, J =10.5, 4.0 Hz, 1H), 3.55 (dd, half ABX pattern, J=10.3, 5.3 Hz, 1H), 2.63 (dddd, J=15, 12, 7.5, 1 Hz, 1H), 2.56-2.47 (m, 1H), 2.40-2.32 (m, 2H).

Step 3. Synthesis of cis-2-[(benzyloxy)methyl]-N-(2,4-dimethoxybenzyl)tetrahydro-2H-pyran-4-amine (P2).

1-(2,4-Dimethoxyphenyl)methanamine (23 g; 140 mmol) was added to a solution of compound C6 (20 g; 91 mmol) in methanol (275 ml). The reaction mixture was stirred at room temperature (20°C) for 24 hours, after which it was cooled to -78°C and treated dropwise with lithium borohydride (2 M solution in tetrahydrofuran; 46.0 ml; 92.0 mmol). The reaction mixture was left to slowly warm to room temperature and then stirred at room temperature overnight. This was combined with the same reaction mixture prepared using compound C6 (16.18 g, 73.5 mmol) and concentrated in vacuo. The residue was mixed with saturated aqueous sodium bicarbonate (300 ml) and water (200 ml) and extracted with ethyl acetate (4×200 ml). The combined organic layers were dried over sodium sulfate, filtered, concentrated under reduced pressure, and purified by silica gel chromatography (gradient: 0% to 9% methanol in dichloromethane) to give the product as a light yellow oil. Total yield: 52.0 g; 140 mmol; 85%. LCMS m/z 371.9 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.38-7.25 (m, 5H), 7.12 (d, J=8.0 Hz, 1H), 6.46 (d, half AB quartet, J=2.5 Hz, 1H ), 6.43 (dd, half ABX pattern, J=8.0, 2.5 Hz, 1H), 4.58 (AB quartet, J _AB =12.0 Hz, Δv _AB =23.2 Hz, 2H), 4.07 ( ddd, J=11.5, 4.5, 1.5 Hz, 1H), 3.81 (s, 3H), 3.80 (s, 3H), 3.75 (s, 2H), 3.59-3.39 (m, 4H), 2.75-2.65 (m, 1H), 1.91-1.80 (m, 2H), 1.48-1.35 (m, 1H), 1.23-1.12 (m, 1H).

Getting R3

N ⁴ -{cis-2-[(Benzyloxy)methyl]tetrahydro-2H-pyran-4-yl}-6-chloroquinolin-3,4-diamine (P3)

Stage 1. Synthesis of 4,6-dichloro-3-nitroquinoline (C7).

N,N-dimethylformamide (3.1 ml; 40 mmol) and thionyl chloride (97%; 6.9 ml; 93 mmol) were added to a suspension of 6-chloro-3-nitroquinolin-4-ol (15.38 g; 68. 48 mmol) in dichloromethane (140 ml) and the reaction mixture was heated to reflux. After 5 hours, it was cooled to room temperature, diluted with more dichloromethane (25 ml) and poured into saturated aqueous sodium bicarbonate (250 ml). The aqueous layer was extracted with dichloromethane (100 ml), then passed through a pad of diatomaceous earth, which was then washed with dichloromethane (50 ml). The combined organic layers and organic filtrate were dried over magnesium sulfate, filtered and concentrated in vacuo to give the product as a tan solid. Yield: 16.8 g; quantitative. ¹ H NMR (400 MHz, CDCl ₃ ) δ 9.25 (s, 1H), 8.42 (d, J=2.2 Hz, 1H), 8.17 (d, J=8.9 Hz, 1H), 7.89 (dd, J=9.0, 2.2 Hz, 1H).

Step 2. Synthesis of N-{cis-2-[(benzyloxy)methyl]tetrahydro-2H-pyran-4-yl}-6-chloro-N-(2,4-dimethoxybenzyl)-3-nitroquinoline-4-amine ( C8).

Compound C7 (17.2 g; 70.8 mmol) was added slowly to a solution of compound P2 (20.8 g; 56.0 mmol) and N,N-diisopropylethylamine (21.7 g; 168 mmol) in acetonitrile (300 ml ). The reaction mixture was stirred for 16 hours at room temperature (25° C.), after which time LCMS analysis showed conversion to the product: LCMS m/z 578.0 (observed chlorine isotope pattern) [M+H] ⁺ . The reaction mixture was concentrated to half of its original volume, diluted with water (400 ml) and extracted with ethyl acetate (2×300 ml). The combined organic layers were dried over sodium sulfate, filtered, concentrated in vacuo and purified by silica gel chromatography (gradient: 0% to 25% ethyl acetate in petroleum ether) to give the product as a yellow solid. Yield: 26.1 g; 45.2 mmol; yield 81%. ¹ H NMR (400 MHz, CDCl ₃ ) δ 9.02 (s, 1H), 8.22 (d, J=2.5 Hz, 1H), 7.98 (d, J=9.0 Hz, 1H), 7.69 (dd, J=9.0, 2.5 Hz, 1H), 7.36-7.25 (m, 5H), 6.82 (bd d, J=8.5 Hz, 1H), 6.22-6.18 (m, 2H), 4.57 ( AB quartet, J _AB =12.3 Hz, Δv _AB =9.1 Hz, 2Н), 4.40-4.27 (m, 2Н), 4.15-4.07 (m, 1H), 3.83-3.73 (m, 1H), 3.69 (s, 3H), 3.59-3.40 (m, 4H), 3.54 (s, 3H), 2.00-1.91 (m, 3H), 1.78-1.66 (m, 1H).

Step 3 Synthesis of N-{cis-2-[(benzyloxy)methyl]tetrahydro-2H-pyran-4-yl}-6-chloro-3-nitroquinoline-4-amine (C9).

Trifluoroacetic acid (11.8 g; 103 mmol) was slowly added dropwise to a solution of compound C8 (6.00 g; 10.4 mmol) in dichloromethane (50 ml) at 20°C. The reaction mixture was stirred for 1 hour, after which LCMS analysis indicated a conversion to the product: LCMS m/z 427.9 (observed chlorine isotope pattern) [M+H] ⁺ . The mixture was then combined with the reaction mixture after similar conversion of compound C8 (1.95 g; 3.37 mmol) and concentrated in vacuo. The residue was diluted with saturated aqueous sodium bicarbonate (200 ml) and extracted with ethyl acetate (4×100 ml); the combined organic layers were dried over sodium sulfate, filtered and concentrated under reduced pressure to give the product as a yellow solid (6.40 g) which contained some ethyl acetate as determined by ¹ H NMR analysis. Combined solvent corrected yield: 5.69 g; 13.3 mmol; 96%. ¹ H NMR (400 MHz, CDCl ₃ ) δ 9.36 (s, 1H), 9.07 (broad d, J=9.0 Hz, 1H), 8.10 (d, J=2.0 Hz, 1H), 7.97 ( d, J=9.0 Hz, 1H), 7.73 (dd, J=9.0, 2.0 Hz, 1H), 7.38-7.26 (m, 5H), 4.59 (AB quartet, J _AB =12.0 Hz, Δv _AB =7.2 Hz, 2H), 4.34-4.22 (m, 1H), 4.18 (ddd, J=12.0, 4.5, 1.5 Hz, 1H), 3.69-3.62 (m, 1H), 3.62-3.52 (m, 2H), 3.49 (dd, ABC pattern component, J=10.3, 4.3 Hz, 1H), 2.21-2.12 (m, 2H), 1.88-1.76 (m, 1H ), 1.66-1.55 (m, 1H).

Step 4 Synthesis of N ⁴ -{cis-2-[(benzyloxy)methyl]tetrahydro-2H-pyran-4-yl}-6-chloroquinolin-3,4-diamine (P3).

Platinum on carbon (5%; 1.37 g) was added in one portion to a solution of compound C9 (6.0 g; 14 mmol) in tetrahydrofuran (200 ml) at a temperature of 20°C. The reaction mixture was purged with argon, then saturated with hydrogen and stirred under a hydrogen pressure of 50 psi (344.7 kPa) for 3 hours at 20°C. Filtration and concentration of the filtrate under vacuum gave the product as a brown solid. Yield: 5.75 g; 14.4 mmol; quantitative. LCMS m/z 397.8 (observed chlorine isotope pattern) [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.47 (s, 1H), 7.90 (d, J=9.0 Hz, 1H), 7.73 (d, J=2.0 Hz, 1H), 7.39 (dd, J=9.0, 2.0 Hz, 1H), 7.36-7.24 (m, 5H), 4.56 (AB quartet, J _AB =12.3 Hz, ΔvAB=9.9 Hz, 2H), 4.09 (ddd, J=12, 4.5, 1 Hz, 1H), 3.90 (br. s, 2H), 3.57-3.40 (m, 5H), 3.39-3.31 (br. m, 1H), 1.91-1.82 (m, 2H ), 1.66-1.53 (m, 1H), 1.43-1.33 (m, 1H).

Getting R4

3-Amino-4-[(4,4-difluoro-1-methylpyrrolidin-3-yl)amino]quinoline-6-carbonitrile (Р4)

Stage 1. Synthesis of 4-hydroxy-3-nitroquinoline-6-carbonitrile (C10).

This reaction was carried out to obtain two identical batches. A mixture of 6-bromo-3-nitroquinolin-4-ol (25.0 g; 92.9 mmol), potassium hexacyanoferrate(II) trihydrate (13.7 g; 32.4 mmol), 1,1'-bis(diphenylphosphino )ferrocene (5.15 g; 9.29 mmol), sodium carbonate (11.8 g; 111 mmol) and palladium(II) acetate (1.04 g; 4.63 mmol) in N,N-dimethylformamide (350 ml) was heated at 140°C for 16 hours. The reaction mixture was cooled to room temperature and the two batches were combined and filtered through diatomaceous earth. The filter cake was slowly washed with N,N-dimethylformamide (200 ml) and tert-butyl methyl ether (3.0 L), while the filtrate was stirred. A dark solid precipitated from the filtrate during stirring, and the resulting mixture was stirred at 20°C for 15 minutes and then filtered. This second filtrate was concentrated under vacuum to a volume of approximately 40 ml; the residue was diluted with tert-butyl methyl ether (ca. 200 ml) and the resulting yellow precipitate was collected by filtration and then triturated with ethyl acetate (ca. 200 ml). The product was obtained as a dark yellow solid. Total yield: 20 g; 93 mmol; fifty%. LCMS m/z 216.0 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.00 (s, 1H), 8.51 (d, J=2.0 Hz, 1H), 7.83 (dd, J=8.5, 1.5 Hz, 1H ), 7.69 (d, J=8.5 Hz, 1H).

Stage 2. Synthesis of 4-chloro-3-nitroquinoline-6-carbonitrile (C11).

Phosphorus oxychloride (9.85 g, 64.2 mmol) was added to a solution of compound C10 (5.00 g; 23.2 mmol) in N,N-dimethylformamide at a temperature of 15°C (30 ml), and the reaction mixture was stirred at 15°C for within 1.5 hours. It was then poured into ice water (100 ml) and the resulting suspension filtered. The collected solids were dissolved in tetrahydrofuran (100 ml) and filtered through a pad of silica gel. Concentration of the filtrate in vacuo gave the product as a white solid. Yield: 3.10 g; 13.3 mmol; yield 57%. ¹ H NMR (400 MHz, CDCl ₃ ) δ 9.39 (s, 1H), 8.83 (d, J=1.8 Hz, 1H), 8.35 (d, J=8.8 Hz, 1H), 8.10 (dd, J=8.8, 1.8 Hz, 1H).

Step 3. Synthesis of tert-butyl 4-[(6-cyano-3-nitroquinolin-4-yl)amino]-3,3-difluoropyrrolidine-1-carboxylate (C12).

tert-Butyl 4-amino-3,3-difluoropyrrolidine-1-carboxylate (prepared using the method described by D.C. Behenna et al., US Patent Application 2015 0141402 A1, May 21, 2015; 2.30 g; 10.3 mmol) was dissolved in acetonitrile (20 ml). To this solution were added N,N-diisopropylethylamine (2.01 g; 15.5 mmol) and compound C11 (3.04 g; 13.0 mmol) and the reaction mixture was stirred for 14 hours at 20°C. After removal of the volatiles under vacuum, purification by silica gel chromatography (gradient: 9% to 17% tetrahydrofuran in petroleum ether) gave the product as a pale yellow solid. Yield: 3.20 g; 7.63 mmol; 74%. ¹ H NMR (400 MHz, CDCl ₃ ) δ 9.52 (s, 1H), 9.21-9.04 (br. m, 1H), 8.48 (b. s, 1H), 8.20 (d, J=8.8 Hz, 1H ), 8.00 (dd, J=8.6, 1.5 Hz, 1H), 4.88-4.74 (m, 1H), 4.23 (broad dd, J=9.7, 8.8 Hz, 1H), 4.05 -3.89 (broad m, 1H), 3.89-3.75 (m, 1H), 3.60 (ddd, J=11.4, 8.4, 1.3 Hz, 1H), 1.51 (s, 9H).

Step 4. Synthesis of 4-[(4,4-difluoropyrrolidin-3-yl)amino]-3-nitroquinoline-6-carbonitrile (C13).

Trifluoroacetic acid (1 ml) was added to a solution of compound C12 (1.10 g; 2.62 mmol) in dichloromethane (2 ml) at 15°C. The reaction mixture was then stirred for 1 hour at 15° C., after this time LCMS analysis showed conversion to the product: LCMS m/z 320.1 [M+H] ⁺ , the mixture was concentrated in vacuo and neutralized by adding aqueous bicarbonate sodium (60 ml). The resulting mixture was extracted with ethyl acetate (3×50 ml) and the combined organic layers were concentrated under reduced pressure to give the product as a pale yellow solid. Yield: 810 mg; 2.54 mmol; 97%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.19 (s, 1H), 9.00 (s, 1H), 8.68-8.57 (broad m, 1H), 8.13 (br AB quartet, J _AB =8.5 Hz, Δv _AB =48.4 Hz, 2H), 4.61-4.43 (m, 1H), 3.58 (dd, J=12.0, 7.5 Hz, 1H), 3.41-3.28 (m, 1H), 3.26 -3.12 (m, 1H), 3.12 (dd, J=11.8, 7.3 Hz, 1H).

Step 5. Synthesis of 4-[(4,4-difluoro-1-methylpyrrolidin-3-yl)amino]-3-nitroquinoline-6-carbonitrile (С14).

Sodium triacetoxyborohydride (2.15 g; 10.1 mmol) was added to a mixture of compound C13 (810 mg; 2.54 mmol) in acetonitrile (5 ml) at 0°C. An aqueous solution of formaldehyde (37%; 824 mg; 10.2 mmol) was added to the reaction mixture at 0° C. over 20 minutes and stirring was then continued at room temperature for 1 hour, after which time LCMS analysis showed a conversion to product: LCMS m/z 334.1 [M+H] ⁺ . After removal of the solvents by concentration in vacuo, the residue was basified to pH 8 by addition of aqueous sodium bicarbonate, filtered and concentrated under reduced pressure to give the product as a red solid. Yield: 780 mg; 2.34 mmol; 92%. ¹ H NMR (400 MHz, CDCl ₃ ), characteristic peaks: δ 9.59 (broad d, J=8.8 Hz, 1H), 9.48 (s, 1H), 8.55 (broad s, 1H), 8.14 (d , J=8.4 Hz, 1H), 7.96 (dd, J=8.8, 1.3 Hz, 1H), 3.29-3.03 (m, 3H), 2.86 (ddd, J=9.9, 5, 1, 2.0 Hz, 1H), 2.47(s, 3H).

Step 6. Synthesis of 3-amino-4-[(4,4-difluoro-1-methylpyrrolidin-3-yl)amino]quinoline-6-carbonitrile (P4).

Palladium on carbon (10%; 1 g) was added to a solution of compound C14 (3.00 g; 9.00 mmol) in methanol (30 ml) and the reaction mixture was hydrogenated with hydrogen from a balloon for 2 hours at 25°C. It was then filtered through diatomaceous earth, concentrated in vacuo and purified by silica gel chromatography (gradient: 17% to 33% tetrahydrofuran in petroleum ether) to give the product as a pale yellow solid. Yield: 1.30 g; 4.29 mmol; 48%. ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.59 (s, 1H), 8.24 (d, J=1.8 Hz, 1H), 8.03 (d, J=8.8 Hz, 1H), 7.60 (dd, J=8.8, 1.8 Hz, 1H), 4.32-4.19 (m, 1H), 4.09-3.96 (m, 3H), 3.18-2.97 (m, 3H), 2.64 (ddd, J=9.7 , 6.6, 1.8 Hz, 1H), 2.41 (s, 3H).

Getting P5

6-Chloro-N ⁴ -(3,3-difluorotetrahydro-2H-pyran-4-yl)quinoline-3,4-diamine (Р5)

Stage 1. Synthesis of tert-butyl-(trans-3-hydroxytetrahydro-2H-pyran-4-yl)carbamate (C15).

A solution of trans-4-azidotetrahydro-2H-pyran-3-ol (see M. Chini et al., Tetrahedron 1994, 50, 1261-1274) (14.8 g; 103 mmol) and di-tert-butyl dicarbonate (23, 0 g; 105 mmol) in ethyl acetate (345 ml) was added to palladium on carbon (10%, 1.5 g) and the reaction mixture was stirred under a hydrogen pressure of 50 psi (344.7 kPa) at a temperature of 20 °С to 25°С within 22 hours. It was then filtered through diatomaceous earth and the filter pad washed with ethyl acetate and methanol. The combined filtrates were concentrated in vacuo and the residue was triturated once with a mixture of dichloromethane (10 ml) and [9:1, petroleum ether/tetrahydrofuran] (60 ml) to give the product as a white solid. Yield: 15.8 g; 72.7 mmol; 71%. ¹ H NMR (400 MHz, CDCl ₃ ) δ 4.71-4.62 (broad s, 1H), 4.01 (dd, J=11.4 Hz, 1H), 3.98-3.87 (m, 2H), 3.57-3.42 (m , 2H), 3.40 (ddd, J=12, 12, 2 Hz, 1H), 3.13 (dd, J=11.0, 9.5 Hz, 1H), 1.96-1.88 (m, 1H), 1.59-1.47 (m, 1H, estimated; partially covered by water peak), 1.47 (s, 9H).

Stage 2. Synthesis of tert-butyl-(3-oxotetrahydro-2H-pyran-4-yl)carbamate (C16).

A solution of compound C15 (35.1 g; 162 mmol) in dichloromethane (540 ml) was treated with [1,1,1-tris(acetyloxy)-1,1-dihydro-1,2-benziodoxol-3-(1H)- he] (Dess-Martin periodinan; 81.6 g; 192 mmol) and stirred at 25° C. for 18 hours. The reaction mixture was treated with saturated aqueous sodium bicarbonate and saturated aqueous sodium thiosulfate (250 ml); after stirring for 30 minutes the layers were separated and the aqueous layer was extracted twice with dichloromethane (200 ml). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo. Chromatography on silica gel (gradient: 10% to 30% ethyl acetate in petroleum ether) gave the product as a yellow oil (27.95 g) which contained some aroma derived from the oxidizing agent. This substance was transferred directly to the next stage. ¹ H NMR (400 MHz, CDCl ₃ ), product peaks only: δ 5.49-5.38 (broad s, 1H), 4.55-4.42 (m, 1H), 4.08 (AB quartet, J _AB =14.8 Hz, Δv _AB =40.3 Hz, 2H), 4.07-3.99 (m, 1H), 3.89 (ddd, J=12.0, 11.5, 3.0 Hz, 1H), 2.75-2.63 (m, 1H), 1.96-1.81 (m, 1H), 1.44 (s, 9H).

Stage 3. Synthesis of tert-butyl-(3,3-difluorotetrahydro-2H-pyran-4-yl)carbamate (C17).

A solution of compound C16 (from the previous step; 27.95 g) in dichloromethane (124 mL) was slowly added to a suspension of difluoro-4-morpholinylsulfonium tetrafluoroborate (XtalFluor- ^M® ; 39.5 g; 163 mmol) and triethylamine trihydrofluoride (28.6 g; 177 mmol) in dichloromethane (384 ml) at 0°C, and the reaction mixture was left to slowly warm to 25°C. After three days, the reaction mixture was treated with saturated aqueous sodium bicarbonate (500 ml) and extracted with dichloromethane (500 ml). The organic layer was dried over sodium sulfate, filtered and concentrated in vacuo. Chromatography on silica gel (eluent: 10% ethyl acetate in petroleum ether) gave the product as a yellow solid. Yield: 8.93 g; 37.6 mmol; 23% in two stages. ¹ H NMR (400 MHz, CDCl ₃ ) δ 4.91-4.75 (broad m, 1H), 4.18-3.94 (m, 3H), 3.55-3.43 (m, 1H), 3.46 (dd, J=30.4, 12.8 Hz, 1H), 2.07-1.97 (m, 1H), 1.86-1.71 (m, 1H), 1.47 (s, 9H).

Step 4. Synthesis of 3,3-difluorotetrahydro-2H-pyran-4-amine hydrochloride salt (C18).

A solution of hydrogen chloride in methanol (4 M; 16.8 ml; 67.2 mmol) was added to a solution of compound C17 (3.18 g; 13.4 mmol) in methanol (35 ml) at a temperature of 10°C. After the reaction mixture was stirred at 10° C. for 1 hour, it was concentrated under vacuum to give the product as a white solid. Yield: 2.32 g; 13.4 mmol; quantitative. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.03-8.89 (br s, 3H), 4.06-3.57 (m, 4H, assumed; partially overlain by water peak), 3.57-3.47 (m, 1H), 2.20 -2.08 (m, 1H), 1.88-1.72 (m, 1H).

Step 5. Synthesis of 6-chloro-N-(3,3-difluorotetrahydro-2H-pyran-4-yl)-3-nitroquinolin-4-amine (C19).

N,N-Diisopropylethylamine (9.2 ml, 52.8 mmol) was added to a solution of compound C7 (3.2 g, 13.2 mmol) and compound C18 (2.32 g, 13.4 mmol) in acetonitrile (40 ml) at a temperature of 10°C, and the reaction mixture was stirred at 10°C for 16 hours. It was then combined with two additional reaction mixtures prepared using Compound C7 (1.2 g; 4.9 mmol and 90 mg; 0.37 mmol) and concentrated in vacuo. Chromatography on silica gel (gradient: 0% to 20% ethyl acetate in petroleum ether) gave the product as a yellow solid. Total yield: 4.5 g; 13 mmol; 70%. LCMS m/z 344.0 (observed chlorine isotope pattern) [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 9.40 (s, 1H), 8.60 (broad d, J=10.1 Hz, 1H), 8.05 (d, J=1.8 Hz, 1H), 8.05 ( d, J=8.8 Hz, 1H), 7.77 (dd, J=9.2, 2.2 Hz, 1H), 4.40-4.26 (m, 1H), 4.17-4.02 (m, 2H), 3.59 ( broad ddd, J=12, 12, 1 Hz, 1H), 3.48 (dd, J=29.0, 12.8 Hz, 1H), 2.40-2.32 (m, 1H), 2.28-2.16 (m, 1H ).

Stage 6. Synthesis of 6-chloro-N ⁴ -(3,3-difluorotetrahydro-2H-pyran-4-yl)quinolin-3,4-diamine (P5).

A mixture of compound C19 (4.40 g; 12.8 mmol) and platinum on carbon (5%; 250 mg) in tetrahydrofuran (50 ml) was degassed with nitrogen at 20° C. and then subjected to hydrogenation at 50 psi (344 .7 kPa) and 20°C for 2 hours. The reaction mixture was filtered and the filter cake was washed with tetrahydrofuran (3×10 ml). The combined filtrates were concentrated in vacuo, combined with the crude product after a similar reaction using compound C19 (100 mg; 0.29 mmol) and purified by silica gel chromatography (gradient: 0% to 20% methanol in dichloromethane) to give product as a yellow solid. Total Yield: 3.9g; 12.4 mmol; 95%. LCMS m/z 314.1 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.48 (s, 1H), 7.90 (d, J=9.3 Hz, 1H), 7.78 (d, J=2.0 Hz, 1H), 7.41 (dd, J=9.0, 2.2 Hz, 1H), 4.10-4.01 (m, 2H), 3.99-3.93 (broad s, 2H), 3.85-3.69 (m, 2H), 3.51-3.42 (m, 1H ), 3.44 (dd, J=31.3, 12.7 Hz, 1H), 2.10-1.95 (m, 2H).

Getting R6

6-Chloro-N ⁴ -[(4S)-3,3-difluorotetrahydro-2H-pyran-4-yl]quinoline-3,4-diamine (P6)

Step 1: Synthesis of tert-butyl-[(3R,4S)-3-hydroxytetrahydro-2H-pyran-4-yl]carbamate (C20).

(3R,4S)-4-aminotetrahydro-2H-pyran-3-ol solution (see M.A. Brodney et al., in PCT International Patent Application No. WO 2016009297 A1 published January 21, 2016) (383 mg; 3.27 mmol) and di-tert-butyl dicarbonate (714 g; 3.27 mmol) in dichloromethane (33 ml) was treated with triethylamine (0.46 ml; 3.3 mmol) and the reaction mixture was stirred at room temperature overnight . Concentration in vacuo gave the product as a yellowish solid. Yield: 707 mg; 3.25 mmol; 99%. ¹ H NMR (400 MHz, CDCl ₃ ) δ 4.69-4.56 (br s, 1H), 4.02 (br dd, J=11.3, 4.7 Hz, 1H), 3.96-3.86 (m, 2H) , 3.58-3.44 (m, 2H), 3.40 (ddd, J=12.1, 11.7, 2.3 Hz, 1H), 3.13 (dd, J=11.3, 9.4 Hz, 1H), 1.96-1.87 (m, 1H), 1.58-1.48 (m, 1H, inferred; partially overlain by water peak), 1.47 (s, 9H).

Stage 2. Synthesis of tert-butyl-[(4S)-3-oxotetrahydro-2H-pyran-4-yl]carbamate (C21).

A solution of compound C20 (707 mg; 3.25 mmol) in dichloromethane (40 ml) was treated with [1,1,1-tris(acetyloxy)-1,1-dihydro-1,2-benziodoxol-3-(1H)- he] (95%; 1.74 g; 3.90 mmol) and stirred at room temperature for 4 hours. The reaction mixture was quenched with saturated aqueous sodium bicarbonate (50 ml) and saturated aqueous sodium thiosulfate (50 ml) and stirred for 30 minutes. The aqueous layer was extracted twice with dichloromethane and the combined organic layers were dried over magnesium sulfate, filtered and concentrated in vacuo. Chromatography on silica gel (gradient: 20% to 80% ethyl acetate in heptane) gave the product as a white solid. Yield: 546 mg; 2.54 mmol; 78%. GCMS m/z 215.1 [M ⁺ ]. ¹ H NMR (400 MHz, CDCl ₃ ) δ 5.49-5.36 (broad s, 1H), 4.49-4.36 (m, 1H), 4.05 (AB quartet, J _AB =14.6 Hz, Δv _AB =38.1 Hz, 2H), 4.04-3.96 (m, 1H), 3.85 (ddd, J=12.1, 11.3, 3.1 Hz, 1H), 2.70-2.59 (m, 1H), 1.92-1.78 (m , 1H), 1.41 (s, 9H).

Step 3: Synthesis of tert-butyl [(4S)-3,3-difluorotetrahydro-2H-pyran-4-yl]carbamate (C22).

A solution of compound C21 (540 mg; 2.51 mmol) in dichloromethane (5 ml) was added slowly to a suspension of difluoro-4-morpholinylsulfonium tetrafluoroborate (1.22 g; 5.02 mmol) and triethylamine trihydrofluoride (0.9 ml; 5. 5 mmol) in dichloromethane (10 ml) at 0°C. The ice bath was removed and the reaction mixture was stirred at room temperature overnight, then at 40° C. for 90 minutes. After cooling to room temperature, the reaction mixture was carefully treated with saturated aqueous sodium bicarbonate {Caution: gas evolution}. The aqueous layer was extracted twice with dichloromethane and the combined organic layers were washed with water, dried over magnesium sulfate, filtered and concentrated in vacuo. Chromatography on silica gel (gradient: 15% to 45% ethyl acetate in heptane) gave the product as a yellow solid which was used in the next step. By analysis ^{of 1} H NMR, it was found that this substance contained some impurities. GCMS m/z 138.1 {[M-(2-methylprop-1-ene and carbon dioxide)] + H} ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ), product peaks only: δ 4.93-4.81 (m, 1H), 4.16-3.93 (m, 3H), 3.51-3.43 (m, 1H), 3.45 (dd, J=30 .4, 12.9 Hz, 1H), 2.05-1.96 (m, 1H), 1.83-1.71 (m, 1H), 1.45 (s, 9H).

Step 4: Synthesis of (4S)-3,3-difluorotetrahydro-2H-pyran-4-amine hydrochloride salt (C23).

Concentrated hydrochloric acid (2 ml) was added to a solution of compound C22 (from the previous step; not more than 2.51 mmol) in ethanol (10 ml) and the reaction mixture was stirred at room temperature overnight. Removal of solvents under vacuum gave the product as a brown solid. Yield: 155 mg; 0.893 mmol; 36% in two stages. GCMS m/z 137.1 [M ⁺ ]. ¹ H NMR (400 MHz, CD ₃ OD) δ 4.09-3.86 (m, 3H), 3.65 (dd, J=31.2, 12.9 Hz, 1H), 3.65-3.56 (m, 1H), 2.23- 2.14 (m, 1H), 2.03-1.90 (m, 1H).

Step 5: Synthesis of 6-chloro-N-[(4S)-3,3-difluorotetrahydro-2H-pyran-4-yl]-3-nitroquinolin-4-amine (C24).

N,N-Diisopropylethylamine (0.41 ml; 2.4 mmol) was added to a mixture of compound C7 (190 mg; 0.782 mmol) and compound C23 (136 mg; 0.783 mmol) in acetonitrile (3 ml) and the reaction mixture was stirred at 60°C during the night. After cooling to room temperature, the reaction mixture was concentrated in vacuo and partitioned between water and ethyl acetate. A small amount of saturated sodium bicarbonate aqueous solution was added to adjust the aqueous layer to pH 9, and the aqueous layer was extracted twice with ethyl acetate. The combined organic layers were dried over magnesium sulfate, filtered, concentrated in vacuo and purified by silica gel chromatography (gradient: 5% to 35% ethyl acetate in heptane) to give the product as a bright yellow solid. Yield: 164 mg; 0.477 mmol; 61%. LCMS m/z 344.4 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 9.39 (s, 1H), 8.61 (broad d, J=10.2 Hz, 1H), 8.04 (d, J=8.6 Hz, 1H), 8.04 ( d, J=2.3 Hz, 1H), 7.77 (dd, J=9.0, 2.3 Hz, 1H), 4.40-4.26 (m, 1H), 4.17-4.02 (m, 2H), 3.59 ( broaden ddd, J=12, 12, 1.5 Hz, 1H), 3.48 (dd, J=29.1, 12.7 Hz, 1H), 2.40-2.32 (m, 1H), 2.29-2.16 (m , 1H).

Step 6: Synthesis of 6-chloro-N ⁴ -[(4S)-3,3-difluorotetrahydro-2H-pyran-4-yl]quinolin-3,4-diamine (P6).

Zinc dust (97.5%; 312 mg; 4.65 mmol) was added to a suspension of compound C24 (160 mg; 0.466 mmol) in methanol (3 ml) and concentrated ammonium hydroxide solution (3 ml). The reaction mixture was stirred at room temperature for 2 hours, after which it was filtered through diatomaceous earth. The filter pack was washed with dichloromethane and methanol and the combined filtrates were concentrated in vacuo. The residue was diluted with half-saturated aqueous sodium chloride and extracted three times with dichloromethane. The combined organic layers were dried over magnesium sulfate, filtered, concentrated under reduced pressure and purified by silica gel chromatography (gradient: 0% to 100% ethyl acetate in heptane) to give the product as a pale tan oil. Yield: 78 mg; 0.249 mmol; 54%. LCMS m/z 314.4 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.47 (s, 1H), 7.89 (d, J=9.0 Hz, 1H), 7.77 (d, J=2.3 Hz, 1H), 7.39 (dd, J=8.8, 2.1 Hz, 1H), 4.08-3.89 (m, 3H), 3.84-3.68 (m, 2H), 3.49-3.40 (m, 1H), 3.42 (dd, J=31.4 , 12.7 Hz, 1H), 2.07-1.94 (m, 2H).

Getting R7

N ⁴ -[(2R,4R)-2-Methyltetrahydro-2H-pyran-4-yl]-6-(trifluoromethyl)quinoline-3,4-diamine (P7)

Step 1. Synthesis of 3-nitro-6-(trifluoromethyl)quinolin-4-ol (С25).

A solution of 6-(trifluoromethyl)quinolin-4-ol (2.00 g; 9.38 mmol) in concentrated nitric acid (10 ml) was stirred for 14 hours at 50°C, after which it was poured into water (50 ml) . The resulting solid was isolated by filtration to give the product as a pale yellow solid. Yield: 1.80 g; 6.97 mmol; 74%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.29 (s, 1H), 8.46 (s, 1H), 8.11 (d, J=9.0 Hz, 1H), 7.92 (d, J=8.5 Hz, 1H).

Stage 2. Synthesis of 4-chloro-3-nitro-6-(trifluoromethyl)quinoline (C26).

Phosphorus oxychloride (3.25 ml; 34.9 mmol) was added to a solution of compound C25 (3.00 g; 11.6 mmol) in N,N-dimethylformamide (10 ml) at 15° C. and the reaction mixture was stirred at for 2 hours at 15°C. Then it was poured into water (80 ml). Collection of the precipitate by filtration gave the product as a solid (2.40 g). This material contained impurities as determined by ¹ H NMR analysis and was passed directly to the next step. ¹ H NMR (400 MHz, DMSO-d ₆ ), product peaks only: δ 9.22 (s, 1H), 8.40 (br s, 1H), 8.03 (br d, J=8.5 Hz, 1H), 7.92-7.97 (m, 1H).

Stage 3. Synthesis of N-(2,4-dimethoxybenzyl)-N-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-3-nitro-6-(trifluoromethyl)quinoline-4-amine (C27).

N,N-Diisopropylethylamine (3.36 g; 26.0 mmol) and compound P1 (2.43 g; 9.16 mmol) were slowly added to a solution of compound C26 (from the previous step; 2.40 g; not more than 8, 68 mmol) in acetonitrile (30 ml) at 15°C, and the reaction mixture was stirred for 30 minutes at 80°C. Water (100 ml) was added and the resulting mixture was extracted with ethyl acetate (3×100 ml). The combined organic layers were concentrated in vacuo and the residue was purified by silica gel chromatography (gradient: 9% to 25% ethyl acetate in petroleum ether) to give the product as a yellow solid. Yield: 3.40 g; 6.73 mmol; 58% in 2 stages. ¹ H NMR (400 MHz, CDCl ₃ ) δ 9.11 (s, 1H), 8.60 (broad s, 1H), 8.15 (d, J=9.0 Hz, 1H), 7.92 (dd, J=8.8 , 1.8 Hz, 1H), 6.84 (d, J=8.0 Hz, 1H), 6.22 (dd, J=8.3, 2.3 Hz, 1H), 6.16 (d, J=2.0 Hz, 1H), 4.33-4.44 (m, 2H), 4.02-4.10 (m, 1H), 3.77-3.87 (m, 1H), 3.68 (s, 3H), 3.50 (s, 3H), 3.36-3.46 ( m, 2H), 1.95-2.10 (m, 3H), 1.67-1.78 (m, 1H), 1.23 (d, J=6.0 Hz, 3H).

Step 4. Synthesis of N-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-3-nitro-6-(trifluoromethyl)quinoline-4-amine (C28).

Trifluoroacetic acid (7.67 g; 67.3 mmol) was added to a solution of compound C27 (3.40 g; 6.73 mmol) in dichloromethane (30 ml) at 15° C. and the reaction mixture was stirred for 30 minutes at 15°C. The solvents were removed under vacuum and the residue was diluted with water (100 ml) and extracted with ethyl acetate (3×100 ml). The combined organic layers were concentrated in vacuo to give the product (2.0 g) as a pale yellow solid, part of which was used directly in the next step. LCMS m/z 355, [M+H] ⁺ .

Step 5. Synthesis of N ⁴ -[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-6-(trifluoromethyl)quinoline-3,4-diamine (P7).

Iron powder (314 mg; 5.2 mmol) and ammonium chloride (301 mg; 5.3 mmol) were added to a solution of compound C28 (from the previous step; 200 mg; 0.54 mmol) in ethanol (5 ml) and water ( 1 ml), and the reaction mixture was stirred for 1 hour at 80°C. It was then filtered through diatomaceous earth and the filtrate concentrated in vacuo. Chromatography on silica gel (gradient: 9% to 33% ethyl acetate in petroleum ether) gave the product as a pale gray solid. Yield: 140 mg; 0.430 mmol; 80% in 2 stages. LCMS m/z 325.9 [M+H] ⁺ .

Getting R8

3-Amino-4-{[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]amino}quinoline-6-carbonitrile (Р8)

Step 1. Synthesis of 4-{(2,4-dimethoxybenzyl)[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]amino}-3-nitroquinoline-6-carbonitrile (C29).

To a solution of compound C11 (8.81 g; 37.7 mmol) in acetonitrile (80 ml) was added compound P1 (11.0 g; 41.5 mmol) followed by N,N-diisopropylethylamine (5.85 g; 45 .3 mmol). The reaction mixture was stirred for 2 hours at room temperature, after which it was concentrated in vacuo and purified by silica gel chromatography (eluent: 4:1, petroleum ether/ethyl acetate) to give the product as a viscous orange oil which slowly solidified. Yield: 15.0 g; 32.4 mmol; 86%. LCMS m/z 313.0 [M-(2,4-dimethoxybenzyl)+H] ⁺ . ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.18 (s, 1H), 8.55 (bd dd, J=1.3, 1 Hz, 1H), 8.15 (d, J=1.0 Hz, 2H ), 6.88 (d, J=8.0 Hz, 1H), 6.24-6.30 (m, 2H), 4.33 (broad AB quartet, J _AB =14.5 Hz, Δv _AB =12 Hz, 2H), 3.76 -3.92 (m, 2H), 3.62 (s, 3H), 3.42 (s, 3H), 3.3-3.4 (m, 2H, inferred; significantly overlapped by water peak), 1.83-2.00 (m, 2H), 1.70-1.83 (m, 1H), 1.42-1.54 (m, 1H), 1.09 (d, J=6.0 Hz, 3H).

Step 2. Synthesis of 4-{[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]amino}-3-nitroquinoline-6-carbonitrile (C30).

A mixture of compound C29 (15.0 g; 32.4 mmol) and trifluoroacetic acid (18.5 g; 162 mmol) in dichloromethane (150 ml) was stirred at room temperature for 30 minutes, after which it was concentrated to a volume of 20 ml and treated with saturated aqueous sodium bicarbonate (200 ml). The aqueous layer was extracted with dichloromethane (3×150 ml) and the combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo to give the product as a yellow solid. Yield: 5.68 g; 18.2 mmol; 56%. LCMS m/z 313.0 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.06-9.09 (m, 2H), 8.30 (broad d, J=9.0 Hz, 1H), 8.14 (dd, half ABX pattern, J=8, 7, 1.6 Hz, 1H), 8.01 (d, half AB quartet, J=8.8 Hz, 1H), 3.87-3.93 (m, 1H), 3.69-3.82 (m, 1H), 3.3-3.5 ( m, 2H, inferred; significantly overlapped by water peak), 1.87-2.03 (m, 2H), 1.60-1.72 (m, 1H), 1.36-1.47 (m, 1H), 1.11 (d, J=6.0 Hz, 3H).

Step 3. Synthesis of 3-amino-4-{[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]amino}quinoline-6-carbonitrile (P8).

Ethanol (60 ml) and water (15 ml) were added to a mixture of compound C30 (5.68 g; 18.2 mmol), iron (10.2 g; 183 mmol) and ammonium chloride (9.73 g; 182 mmol) . The reaction mixture was heated to 80°C for 1 hour, after which it was diluted with ethanol (100 ml) and filtered. The filtrate was concentrated in vacuo and the resulting solid partitioned between saturated aqueous sodium bicarbonate (100 ml) and dichloromethane (300 ml). The organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure to give the product as a brown solid. Yield: 4.73 g; 16.8 mmol; 92%. LCMS m/z 282.9 [M+H] ⁺ . ¹ H NMR (400 MHz, CD ₃ OD) δ 8.55 (d, J=1.2 Hz, 1H), 8.51 (s, 1H), 7.90 (d, J=8.8 Hz, 1H), 7.60 (dd , J=8.5, 1.8 Hz, 1H), 3.92-4.00 (m, 1H), 3.58-3.69 (m, 1H), 3.39-3.50 (m, 2H), 1.78-1.94 (m, 2H) , 1.56-1.69 (m, 1H), 1.29-1.40 (m, 1H), 1.17 (d, J=6.0 Hz, 3H).

Getting P9

Stage 1. Synthesis of 4-{[(3R)-1-methylpyrrolidin-3-yl]amino}-3-nitroquinoline-6-carbonitrile (С31).

N,N-Diisopropylethylamine (251 mg; 1.94 mmol) was added to a solution of compound C11 (210 mg; 0.899 mmol) and (3R)-1-methylpyrrolidin-3-amine (77.9 mg; 0.778 mmol) in acetonitrile ( 3 ml) at 20°C. The reaction mixture was stirred at 20°C for 2 hours, after which it was concentrated under vacuum. Purification of the residue by silica gel chromatography (gradient: 0% to 1% methanol in dichloromethane) gave the product as a yellow solid. Yield: 210 mg; 0.706 mmol; 91%. LCMS m/z 297.9 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 10.04-10.15 (br. m, 1H), 9.45 (s, 1H), 8.55 (d, J=1.5 Hz, 1H), 8.07 (d, half of AB quartet , J=8.5 Hz, 1H), 7.92 (dd, half ABX pattern, J=8.5, 1.8 Hz, 1H), 4.65-4.74 (m, 1H), 3.02-3.10 (m, 1H) , 2.84-2.90 (m, 1H), 2.80 (dd, half ABX pattern, J=9.9, 5.6 Hz, 1H), 2.61-2.71 (m, 1H) 2.46 (s, 3H), 2.41-2.50 (m, 1H), 2.06-2.16 (m, 1H).

Stage 2. Synthesis of 3-amino-4-{[(3R)-1-methylpyrrolidin-3-yl]amino}quinoline-6-carbonitrile (P9).

Ammonium chloride (36 mg; 0.673 mmol) and iron powder (75.1 mg; 1.34 mmol) were added to a solution of compound C31 (100 mg; 0.336 mmol) in a mixture of ethanol (1 ml) and water (0.25 ml) , and the reaction mixture was stirred at 80°C for 1 hour. It was then filtered and the filter cake was washed with methanol (30 ml). The organic layer from the combined filtrates was concentrated in vacuo and purified by silica gel chromatography (gradient: 0% to 15% methanol in dichloromethane) to give the product as a yellow solid. Yield: 112 mg; accepted as quantitative. ¹ H NMR (400 MHz, DMSO-d ₆ ), characteristic peaks: δ 8.65-8.71 (br.s, 1H), 8.58 (s, 1H), 7.89 (d, J=8.5 Hz, 1H), 7.62 (dd, J=8.5, 2.0 Hz, 1H), 5.56-5.70 (br.s, 1H), 5.43 (d, J=10.5 Hz, 1H), 4.32-4.46 (br.m, 1Н), 2.81 (s, 3Н), 1.84-1.95 (m, 1H).

Getting P10

Stage 1. Synthesis of 6-chloro-N-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-3-nitroquinolin-4-amine (С32).

Compound C7 (12.2 g, 50.2 mmol) was added to a solution of compound P1 (13.3 g, 50.1 mmol) and N,N-diisopropylethylamine (13.1 ml, 75.2 mmol) in acetonitrile (250 ml) and the reaction mixture was heated to 55° C. overnight. After concentration in vacuo, the residue was partitioned between aqueous sodium bicarbonate (100 ml) and dichloromethane (150 ml). The aqueous layer was extracted with dichloromethane (2×50 ml) and the combined organic layers were treated with trifluoroacetic acid (25 ml). {Caution: exothermic effect!}. After 20 minutes, a saturated aqueous sodium carbonate solution (150 ml) was added in portions and the mixture was allowed to stir for 10 minutes. The aqueous layer was extracted twice with dichloromethane and the combined organic layers concentrated in vacuo to give a reddish solid (17.3 g); it was triturated with diethyl ether (230 ml) to give a yellow solid (14.0 g). A portion of this solid (10 g) was purified by supercritical fluid chromatography (column: Lux Amylose-2, 5 μm; mobile phase: 65:35, carbon dioxide/methanol) to give the product as a crystalline solid. The specified absolute configuration was determined by single crystal X-ray diffraction analysis of this substance: see below. Yield: 7.1 g; 22 mmol; 62% (yield corrected for material not purified). ¹ H NMR (400 MHz, CDCl ₃ ) δ 9.36 (s, 1H), 9.11 (broad d, J=9 Hz, 1H), 8.12 (d, J=2.0 Hz, 1H), 7.98 (d, J=8.9Hz, 1H), 7.73(dd, J=8.9, 2.2Hz, 1H), 4.21-4.33(m, 1H), 4.08-4.15(m, 1H), 3.50-3.60( m, 2Н), 2.11-2.22 (m, 2Н), 1.77 (dddd, J=12, 12, 12.5 Hz, 1H), 1.49 (ddd, J=12, 12, 11 Hz, 1H), 1.28 ( d, J=6.2 Hz, 3H).

Single-crystal X-ray diffraction analysis of compound С32

X-ray analysis of single crystals

Data collection was carried out on a Bruker APEX diffractometer at room temperature. Data collection consisted of omega and phi scans.

The structure was determined by direct methods using the SHELX software package in the space group P2 ₁ 2 ₁ 2 ₁ . The structure was then refined using the least squares method in the full matrix approximation. All non-hydrogen atoms were identified and refined using anisotropic displacement parameters.

The hydrogen atom located on nitrogen was identified using the difference Fourier map and refined using limited distances. The rest of the hydrogen atoms were placed in the calculated positions and rotated about their carrier atoms. The final refinement included isotropic displacement parameters for all hydrogen atoms.

Absolute structure analysis using likelihood methods (Hooft, 2008) was performed using the PLATON program (Spek, 2003). The results show that the absolute structure is well defined. The method calculates that the probability that the structure is correct is 100.0. The Huft parameter is reported as 0.017 with a marginal studentized deviation (ESD) of 0.09.

The final R-factor was 4.8%. The final difference Fourier map showed no gaps or shifts in the electron density.

The relevant crystal summary, data acquisition, and refinement parameters are listed in Table A. Atomic coordinates, bond lengths, bond angles, and bias parameters are listed in Tables B-E.

Software and bibliography

SHELXTL, Version 5.1, Bruker AXS, 1997.

PLATON, A.L. Spek, J. Appl. Cryst. 2003, 36, 7-13.

MERCURY, C.F. Macrae, P.R. Edington, P. McCabe, E. Pidcock, G.P. Shields, R. Taylor, M. Towler, and J. van de Streek, J. Appl. Cryst. 2006, 39, 453-457.

OLEX2, O.V. Dolomanov, L.J. Bourhis, R.J. Gildea, J.A.K. Howard, and H. Puschmann, J. Appl. Cryst. 2009, 42, 339-341.

R.W.W. Hooft, L.H. Straver, and A.L. Spek, J. Appl. Cryst. 2008, 41, 96-103.

H.D. Flack, Acta Cryst. 1983, A39, 867-881.

Stage 2. Synthesis of 6-chloro-N ⁴ -[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]quinolin-3,4-diamine (P10).

Zinc dust (97.5%; 12.3 g; 183 mmol) was added in one portion to a suspension of compound C32 (7.40 g; 23.0 mmol) in methanol (100 ml) and concentrated ammonium hydroxide (100 ml). After 1 hour the reaction mixture was filtered through diatomaceous earth, the filter pad was washed with dichloromethane (70 ml). The combined filtrates were diluted with water and the aqueous layer was extracted with dichloromethane (2×60 ml). The combined organic layers were dried over sodium sulfate, filtered, concentrated in vacuo and purified by silica gel chromatography (gradient: 40% to 100% ethyl acetate in heptane) to give the product. Yield: 3.68 g; 12.6 mmol; 55%. ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.48 (s, 1H), 7.91 (d, J=8.9 Hz, 1H), 7.74 (d, J=2.2 Hz, 1H), 7.40 (dd, J=8.9, 2.2 Hz, 1H), 4.02 (brd dd, J=12.5 Hz, 1H), 3.88 (br s, 2H), 3.29-3.56 (m, 4H), 1.82- 1.96 (m, 2H), 1.56 (dddd, J=12.12, 12.5 Hz, 1H), 1.21-1.31 (m, 1H), 1.21 (d, J=6.2 Hz, 3H).

Getting P11

Stage 1. Synthesis of 6-chloro-N-(2,4-dimethoxybenzyl)-N-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-3-nitroquinolin-4-amine (С33) .

Compound C7 (4.00 g, 16.5 mmol), and the reaction mixture was heated at 50°C for 16 hours. It was then concentrated in vacuo and the residue partitioned between ethyl acetate (100 ml) and saturated aqueous sodium bicarbonate (100 ml), after which the aqueous layer was extracted with ethyl acetate (2×150 ml) and the combined organic layers washed with saturated aqueous sodium chloride ( 150 ml), dried over sodium sulfate, filtered and concentrated under reduced pressure. Chromatography on silica gel (gradient: 0% to 80% ethyl acetate in heptane) gave the product as an orange solid. Yield: 6.00 g; 12.7 mmol; 86%. LCMS m/z 472.5 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 9.02 (s, 1H), 8.24 (d, J=2.4 Hz, 1H), 7.98 (d, J=9.0 Hz, 1H), 7.69 (dd, J=9.0, 2.4 Hz, 1H), 6.83 (d, J=8.2 Hz, 1H), 6.24-6.18 (m, 2H), 4.34 (brs, 2H), 4.08-4.00 ( m, 1H), 3.82-3.70 (m, 1H), 3.69 (s, 3H), 3.55 (s, 3H), 3.49-3.38 (m, 2H), 2.02-1.85 (m, 3H), 1.66-1.52 ( m, 1H, assumed; partially covered by water peak), 1.21 (d, J=6.3 Hz, 3H).

Step 2. Synthesis of 2-(4-{(2,4-dimethoxybenzyl)[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]amino}-3-nitroquinolin-6-yl)-2, 2-difluoro-1-phenylethanone (C34).

Chloro[(di(1-adamantyl)-N-butylphosphine)-2-(2-aminobiphenyl)]palladium(II) ( ^cataCXium® A Pd G2; 85.0 mg ; 0.127 mmol), compound C33 (3.00 g; 6.36 mmol) and potassium orthophosphate monohydrate (5.86 g; 25.4 mmol). The tube was then evacuated and filled with nitrogen. This pumping cycle was repeated twice, after which a solution of 2,2-difluoro-1-phenylethanone (1.68 ml; 12.7 mmol) in toluene (37 ml) was added and the reaction mixture was heated at 110° C. for 24 hours. After cooling to room temperature, the reaction mixture was partitioned between saturated aqueous ammonium chloride (250 ml) and ethyl acetate (250 ml). The organic layer was dried over sodium sulfate, filtered, concentrated in vacuo and purified by silica gel chromatography (gradient: 0% to 100% ethyl acetate in heptane) to give the product as an orange solid. Yield: 2.07 g; 3.50 mmol; 55%. LCMS m/z 592.3 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 9.07 (s, 1H), 8.49 (s, 1H), 8.17 (d, half AB quartet, J=8.6 Hz, 1H), 8.08-7.99 (m, 3H ), 7.65 (dd, J=8.7 Hz, 1H), 7.50 (dd, J=8.7 Hz, 2H), 6.83 (d, J=8.2 Hz, 1H), 6.20 (broad d, J=8.6 Hz, 1H), 6.15 (s, 1H), 4.35 (br.s, 2H), 4.04-3.96 (m, 1H), 3.81-3.70 (m, 1H), 3.69 (s, 3H) , 3.47 (s, 3H), 3.30-3.18 (m, 2H), 2.07-1.87 (m, 3H), 1.76-1.64 (m, 1H), 1.21 (d, J=6.3 Hz, 3H).

Stage 3. Synthesis of 6-(difluoromethyl)-N-(2,4-dimethoxybenzyl)-N-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-3-nitroquinolin-4-amine ( C35).

Potassium hydroxide (1.97 g; 35.1 mmol) and water (1.22 ml; 67.7 mmol) were added to a solution of compound C34 (2.0 g; 3.38 mmol) in toluene (20 ml) and the resulting biphasic reaction mixture was vigorously stirred at 100° C. for 11 hours. An aliquot of the reaction mixture was partitioned between saturated aqueous sodium bicarbonate and ethyl acetate; LCMS analysis of the organic layer indicated the presence of both starting material and product. The reaction mixture was cooled to room temperature and partitioned between saturated aqueous sodium bicarbonate (125 ml) and ethyl acetate (150 ml). The organic layer was dried over sodium sulfate, filtered, concentrated in vacuo and subjected to silica gel chromatography (gradient: 0% to 100% ethyl acetate in heptane), which failed to separate compound C35 from compound C34. The isolated mixture was re-placed under reaction conditions for 24 hours, then treated in a similar manner; the crude residue (again a mixture of compounds C35 and C34) was again placed under the initial reaction conditions, this time for 48 hours. The reaction mixture was cooled to room temperature and partitioned between saturated aqueous sodium bicarbonate (125 ml) and ethyl acetate (150 ml). The organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure to give an oily orange residue (1.85 g) which contained both C35 and C34 as shown by LCMS analysis. This substance was used directly in step 4. LCMS m/z 488.5 [M+H] ⁺ .

Improved conversion of compound C34 to 6-(difluoromethyl)-N-(2,4-dimethoxybenzyl)-N-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-3-nitroquinolin-4-amine (C35).

Potassium hydroxide (267 mg; 4.76 mmol) was added to a solution of compound C34 (470 mg; 0.794 mmol) in toluene (4.7 ml) and water (0.28 ml; 16 mmol). The reaction mixture was heated to 100°C for 24 hours, after which it was cooled to room temperature and partitioned between water (150 ml) and dichloromethane (150 ml). The aqueous layer was extracted with dichloromethane (2×100 ml) and the combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo. Chromatography on silica gel (gradient: 0% to 80% ethyl acetate in heptane) gave the product as an orange solid. Yield: 337 mg; 0.691 mmol; 87%. ¹ H NMR (400 MHz, CDCl ₃ ) δ 9.10 (s, 1H), 8.45 (br.s, 1H), 8.14 (d, J=8.6 Hz, 1H), 7.83 (br.dd, J=8 .6, 1Hz, 1H), 6.86 (d, J=8.2Hz, 1H), 6.81 (t, JHF= _56.3Hz , 1H), 6.23 (dd, half ABX pattern, J=8, 2, 2.4 Hz, 1H), 6.17 (d, half AB quartet, J=2.4 Hz, 1H), 4.38 (broad AB quartet, J _AB =14 Hz, Δv _AB =8 Hz, 2H), 4.08-4.02 (m, 1H), 3.88-3.78 (m, 1H), 3.69 (s, 3H), 3.49 (s, 3H), 3.46-3.36 (m, 2H), 2.07-1.94 (m, 3H), 1.73-1.62 (m, 1H), 1.22 (d, J=6.3 Hz, 3H).

Step 4. Synthesis of 6-(difluoromethyl)-N-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-3-nitroquinolin-4-amine (C36).

A solution of compounds C35 and C34 (from step 3; 1.85 g; not more than 3.38 mmol) in dichloromethane (25 ml) was cooled to 0°C and treated with trifluoroacetic acid (1.16 ml; 15.1 mmol). The reaction mixture was allowed to warm to room temperature and stirred at room temperature for 20 minutes, after which it was cooled to 0°C, diluted with dichloromethane (20 ml) and basified to pH 8 by adding saturated aqueous sodium bicarbonate (100 ml). The aqueous layer was extracted with dichloromethane (2×50 ml) and the combined organic layers were washed with saturated aqueous sodium chloride (150 ml), dried over magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (gradient: 0% to 100% ethyl acetate in heptane) followed by trituration with diethyl ether (50 ml) to give the product as a yellow solid. Yield in two stages: 0.70 g; 2.1 mmol; 62%. LCMS m/z 338.3 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 9.44 (s, 1H), 9.34 (broad d, J=7.8 Hz, 1H), 8.37 (s, 1H), 8.11 (d, J=8.6 Hz, 1H), 7.86 (d, J=8.6 Hz, 1H), 6.84 (t, J _HF =56.5 Hz, 1H), 4.39-4.26 (m, 1H), 4.18-4.08 (m, 1H ), 3.61-3.48 (m, 2H), 2.27-2.12 (m, 2H), 1.89-1.74 (m, 1H). 1.58-1.46 (m, 1H), 1.28 (d, J=6.3 Hz, 3H).

Step 5. Synthesis of 6-(difluoromethyl)-N ⁴ -[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]quinolin-3,4-diamine (P11).

A Parr reactor was charged with a solution of compound C36 (0.70 g; 2.1 mmol) in tetrahydrofuran (150 ml) followed by a charge of platinum on carbon (5%; 600 mg). The mixture was purged with nitrogen three times, refilled with hydrogen, after which it was hydrogenated for 2 hours at 30 psi (206.8 kPa). The reaction mixture was then diluted with tetrahydrofuran (50 ml) and filtered through a pad of diatomaceous earth. The filter pack was washed with tetrahydrofuran (3×50 ml) and the combined filtrates were concentrated in vacuo, dissolved in dichloromethane (15 ml) and filtered through an Acrodisc® filter. The filtrate was concentrated under reduced pressure to give the product as a dark brown solid. The product contained some impurities based on ¹ N NMR analysis. Yield: 547 mg; 1.78 mmol; 85%. LCMS m/z 308.4 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ), characteristic peaks: δ 8.56 (s, 1H), 8.06 (d, J=8.6 Hz, 1H), 7.93 (br.s, 1H), 7.57 (br.d , J=8.6 Hz, 1H), 6.83 (t, J _HF =56.3 Hz, 1H), 4.03 (ddd, J=11.7, 4.7, 1.6 Hz, 1H), 3.87 ( br.s, 2Н), 3.61-3.49 (m, 1Н), 3.49-3.39 (m, 2Н), 1.98-1.90 (m, 1Н), 1.90-1.82 (m, 1Н), 1.63-1.51 (m, 1Н ), 1.21 (d, J=6.3 Hz, 3H).

Getting P12

Stage 1. Synthesis of tert-butyl-3,3-difluoro-4-[(6-fluoro-3-nitroquinolin-4-yl)amino]pyrrolidine-1-carboxylate (С37).

N,N-diisopropylethylamine (6.84 g, 52.9 mmol) followed by the addition of tert-butyl-4-amino-3,3-difluoropyrrolidine-1-carboxylate (obtained using the method described by D.C. Behenna et al., in US patent application 20150141402 A1 May 21, 2015; 9.81 g, 44.1 mmol). The reaction mixture was stirred at 20° C. for 48 hours after which it was concentrated in vacuo and purified by silica gel chromatography (gradient: 9% to 17% tetrahydrofuran in petroleum ether) to give the product as a pale yellow solid. Yield: 16.8 g; 40.7 mmol; 92%. ¹ H NMR (400 MHz, CDCl ₃ ) δ 9.39 (s, 1H), 8.87-8.69 (br.m, 1H), 8.13 (dd, J=9.5, 5.5 Hz, 1H), 7.79-7.70 (br.d, J=8 Hz, 1H), 7.63 (ddd, J=9.0, 7.5, 2.5 Hz, 1H), 4.87-4.71 (br.m, 1H), 4.31-4.09 ( br.m, 1H), 4.04-3.84 (br.m, 1H), 3.84-3.69 (m, 1H), 3.63-3.51 (br.m, 1H), 1.50 (s, 9H).

Stage 2. Synthesis of N-(4,4-difluoropyrrolidin-3-yl)-6-fluoro-3-nitroquinoline-4-amine (С38).

Trifluoroacetic acid (50 ml) was added to a solution of compound C37 (16.8 g; 40.7 mmol) in dichloromethane (100 ml) at 15°C and the reaction mixture was stirred for 3 hours at 15°C. At this point, LCMS analysis indicated product formation (LCMS m/z 313.1 [M+H] ⁺ ) and the reaction mixture was concentrated in vacuo. The residue was taken up in aqueous sodium bicarbonate (200 ml) and extracted with ethyl acetate (3×150 ml). Concentration of the combined organic layers under reduced pressure gave the product as a pale yellow solid. Yield: 12.5 g; 40.0 mmol; 98%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.07 (s, 1H), 8.30 (bd, J=9.2 Hz, 1H), 8.26 (dd, J=10.6, 2.6 Hz , 1H), 8.04 (dd, J=9.0, 5.9 Hz, 1H), 7.85-7.78 (m, 1H), 4.53-4.39 (m, 1H), 3.58 (dd, J=11.9, 7.5 Hz, 1H), 3.39-3.25 (m, 1H), 3.24-3.09 (m, 1H), 3.08 (dd, J=11.9, 7.5 Hz, 1H).

Step 3 Synthesis of N-(4,4-difluoro-1-methylpyrrolidin-3-yl)-6-fluoro-3-nitro-quinoline-4-amine (C39).

Sodium triacetoxyborohydride (33.9 g; 160 mmol) was added to a mixture of compound C38 (12.5 g; 40.0 mmol) in acetonitrile (150 ml) at 0°C. An aqueous solution of formaldehyde (37%; 13.0 g; 160 mmol) was slowly added over 20 minutes and the reaction mixture was stirred at room temperature for 1 hour, at which point LCMS analysis indicated that the reaction was complete (LCMS m/z 327 ,1 [M+H ⁺ ]). After the reaction mixture was concentrated to dryness, the residue was basified to pH 8 by adding aqueous sodium bicarbonate. The resulting solid was collected by filtration to give the product as a red solid. Yield: 11.8 g; 36.2 mmol; 90%. ¹ H NMR (400 MHz, CDCl ₃ ), characteristic peaks: δ 9.35 (s, 1H), 9.22 (broad d, J=9.2 Hz, 1H), 8.07 (dd, J=9.0.5, 5 Hz, 1H), 7.83 (dd, J=10.1, 2.6 Hz, 1H), 7.58 (ddd, J=9.2, 7.5, 2.6 Hz, 1H), 3.27 (dd, J=9.7, 6.2 Hz, 1H), 3.15-3.05 (m, 2H), 2.79 (ddd, J=9.9, 5.9, 2.0 Hz, 1H), 2.45 (s, 3H ).

Step 4 Synthesis of N ⁴ -(4,4-difluoro-1-methylpyrrolidin-3-yl)-6-fluoroquinolin-3,4-diamine (P12).

Palladium on carbon (10%; 3.85 g) was added to a solution of compound C39 (11.8 g; 36.2 mmol) in methanol (100 ml) and the resulting mixture was hydrogenated (30 psi (206.8 kPa)) at 25°C for 1 hour. This reaction mixture was combined with a similar reaction mixture prepared using compound C39 (3.60 g, 11.0 mmol) and filtered through diatomaceous earth. The filtrate was concentrated in vacuo and purified using silica gel chromatography (gradient: 9% to 17% tetrahydrofuran in petroleum ether). The product was obtained as a pale yellow solid. Total yield: 8.40 g; 28.3 mmol; 60%. LCMS m/z 291.1 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.45 (s, 1H), 7.95 (dd, J=9.0, 5.5 Hz, 1H), 7.43 (dd, J=10.6, 2.6 Hz , 1H), 7.23 (ddd, J=9.0, 8.1, 2.6 Hz, 1H), 4.26-4.12 (m, 1H), 4.05-3.89 (br. s, 2H), 3.79 (br. d, J=11.0 Hz, 1H), 3.23-2.93 (m, 3H), 2.63-2.55 (m, 1H), 2.38 (s, 3H).

Getting P13

Step 1. Synthesis of tert-butyl 4-[(6-chloro-3-nitroquinolin-4-yl)amino]-3,3-difluoropyrrolidine-1-carboxylate (С40).

To a solution of compound C7 (13.1 g; 53.9 mmol) in acetonitrile (60 mL) was added N,N-diisopropylethylamine (11.3 mL; 64.9 mmol) followed by a solution of tert-butyl-4-amino- 3,3-difluoropyrrolidine-1-carboxylate (prepared using the method described by D.C. Behenna et al. in US Patent Application 2015 0141402 A1 May 21, 2015; 12.0 g; 54.0 mmol) in acetonitrile (5 ml). After the reaction mixture was stirred at 20°C for 32 hours, it was diluted with water (100 ml). The resulting solid was collected by filtration and purified by silica gel chromatography (gradient: 0% to 25% tetrahydrofuran in petroleum ether) to give the product as a yellow solid. Yield: 12.0 g; 28.0 mmol; 52%. LCMS m/z 428.7 (observed chlorine isotope pattern) [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 9.41 (s, 1H), 8.91-8.78 (br.m, 1H), 8.08 (ynmp.s, 1H), 8.06 (d, J=9.0 Hz, 1H ), 7.79 (dd, J=9.0, 2.0 Hz, 1H), 4.86-4.72 (br.m, 1H), 4.30-4.12 (br.m, 1H), 4.03-3.86 (br.m, 1H), 3.86-3.71 (m, 1H), 3.64-3.52 (br. m, 1H), 1.51 (s, 9H).

Stage 2. Synthesis of 6-chloro-N-(4,4-difluoropyrrolidin-3-yl)-3-nitroquinolin-4-amine (C41).

Trifluoroacetic acid (60 ml) was added to a solution of compound C40 (11.9 g; 27.8 mmol) in dichloromethane (100 ml) and the reaction mixture was stirred at 20° C. for 12 hours. The solvents were then removed by concentration in vacuo and the residue was carefully basified by the addition of aqueous sodium bicarbonate (500 ml). The resulting mixture was extracted with 2-methyltetrahydrofuran (2×200 ml) and the combined organic layers were dried over sodium sulfate, filtered and concentrated under reduced pressure to give the product as a yellow solid (10.9 g) which was used in the next step. LCMS m/z 328.5 (observed chlorine isotope pattern) [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.08 (s, 1H), 8.57 (s, 1H), 8.42-8.29 (br s, 1H), 7.94 (br AB quartet, J _AB =8 Hz , Δv _AB =26 Hz, 2H), 4.45-4.30 (br. m, 1H), 3.57-3.46 (b. m, 1H), 3.33-3.22 (m, 1H, assumed; partially covered by water peak), 3.21- 2.98 (m, 3H).

Stage 3. Synthesis of 6-chloro-N-(4,4-difluoro-1-methylpyrrolidin-3-yl)-3-nitroquinoline-4-amine (С42).

Sodium triacetoxyborohydride (26.8 g; 126 mmol) was added to a solution of compound C41 (from the previous step; 10.4 g; not more than 26.5 mmol) in acetonitrile (110 ml) at 0°C. An aqueous solution of formaldehyde (37%; 10.3 g; 127 mmol) was added over 20 minutes and the reaction mixture was stirred at room temperature for 1 hour. It was then combined with a similar reaction mixture obtained using compound C41 (from the previous step; 500 mg; not more than 1.27 mmol) and concentrated in vacuo. The residue was basified to pH 8 by adding aqueous sodium bicarbonate and the resulting solid was collected by filtration to give the product as a red solid. Total yield: 8.60 g; 25.1 mmol; 90% in two stages. LCMS m/z 342.6 (observed chlorine isotope pattern) [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 9.38 (s, 1H), 9.30 (broad d, J=9.2 Hz, 1H), 8.18 (d, J=2.2 Hz, 1H), 8.01 ( d, J=8.8 Hz, 1H), 7.75 (dd, J=9.2, 2.2 Hz, 1H), 4.83-4.71 (m, 1H), 3.27 (ddd, J=10.1, 6 .2, 0.9 Hz, 1H), 3.16-3.07 (m, 2H), 2.81 (ddd, J=9.9, 5.7, 2.0 Hz, 1H), 2.46 (s, 3H).

Stage 4. Synthesis of 6-chloro-N ⁴ -(4,4-difluoro-1-methylpyrrolidin-3-yl)quinolin-3,4-diamine (P13).

Platinum(IV) oxide (5.0 g; 22 mmol) was added to a solution of compound C42 (8.50 g; 24.8 mmol) in methanol (100 ml) and the resulting mixture was hydrogenated at 25° C. for 4 hours at use of hydrogen from a cylinder. The reaction mixture was combined with a similar reaction mixture prepared using compound C42 (100 mg; 0.292 mmol), filtered through diatomaceous earth and concentrated in vacuo. Chromatography on silica gel (gradient: 17% to 100% tetrahydrofuran in petroleum ether) gave the product as a brown oil which solidified on standing overnight. Total yield: 5.02 g; 16.1 mmol; 64%. LCMS m/z 312.9 (observed chlorine isotope pattern) [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.48 (s, 1H), 7.90 (d, J=9.0 Hz, 1H), 7.81 (d, J=2.0 Hz, 1H), 7.41 (dd, J=8.8, 2.3 Hz, 1H), 4.29-4.16 (m, 1H), 3.95 (br. s, 2H), 3.86 (br. d, J=11.0 Hz, 1H), 3.19- 2.96 (m, 3H), 2.61 (ddd, J=9.7.2 Hz, 1H), 2.41 (s, 3H).

Example 1 and 2

[(2S,4R)-4-(8-Chloro-2-ethyl-1H-imidazo[4,5-c]quinolin-1-yl)tetrahydro-2H-pyran-2-yl]acetonitrile (1) and [ (2R,4S)-4-(8-chloro-2-ethyl-1H-imidazo[4,5-c]quinolin-1-yl)tetrahydro-2H-pyran-2-yl]acetonitrile (2)

Stage 1. Synthesis of 1-{cis-2-[(benzyloxy)methyl]tetrahydro-2H-pyran-4-yl}-8-chloro-2-ethyl-1H-imidazo[4,5-c]quinoline (С43) .

A solution of compound P3 (800 mg; 2.01 mmol) in propionic acid (10 ml) and 1,1,1-triethoxypropane (10 ml) was stirred at 110°C for 2.5 hours, after which it was combined with a similar reaction mixture prepared using compound P3 (100 mg; 0.251 mmol) and poured into water. The resulting mixture was neutralized with solid potassium carbonate and extracted with ethyl acetate (2×100 ml). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo. Purification by silica gel chromatography (gradient: 0% to 2% methanol in dichloromethane) gave the product as a yellow solid. Yield: 875 mg; 2.01 mmol; 89%. LCMS m/z 436.1 [M+H] ⁺ .

Stage 2. Synthesis of [cis-4-(8-chloro-2-ethyl-1H-imidazo[4,5-c]quinolin-1-yl)tetrahydro-2H-pyran-2-yl]methanol (C44).

A solution of compound C43 (875 mg; 2.01 mmol) in dichloromethane (17 ml) at 0° C. was treated with boron trichloride (1 M solution; 6.02 ml; 6.02 mmol) and the reaction mixture was stirred at 20° C. for 2 hours, after which it was poured into an aqueous solution of sodium bicarbonate (50 ml) and was extracted with ethyl acetate (2×50 ml). The combined organic layers were dried over sodium sulfate, filtered, concentrated under reduced pressure and purified by silica gel chromatography (gradient: 0% to 2.8% methanol in dichloromethane) to give the product as a yellowish foamy solid. Yield: 490 mg; 1.42 mmol; 71%. LCMS m/z 346.0 [M+H] ⁺ .

Stage 3. Synthesis of [cis-4-(8-chloro-2-ethyl-1H-imidazo[4,5-c]quinolin-1-yl)tetrahydro-2H-pyran-2-yl]methylmethanesulfonate (C45).

Triethylamine (430 mg, 4.25 mmol) and methanesulfonyl chloride (195 mg, 1.70 mmol) were added to a solution of compound C44 (490 mg; 1.42 mmol) in dichloromethane (10 ml) at 0°C. The reaction mixture was stirred at 20°C for 1 hour, after which it was poured into water (50 ml) and extracted with ethyl acetate (2×50 ml). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo to give the product as a yellow foamy solid (640 mg) which was passed directly to the next step. LCMS m/z 423.8 (observed chlorine isotope pattern) [M+H] ⁺ .

Step 4. Synthesis of [(2S,4R)-4-(8-chloro-2-ethyl-1H-imidazo[4,5-c]quinolin-1-yl)tetrahydro-2H-pyran-2-yl]acetonitrile ( 1) and [(2R,4S)-4-(8-chloro-2-ethyl-1H-imidazo[4,5-c]quinolin-1-yl)tetrahydro-2H-pyran-2-yl]acetonitrile (2 ).

To a solution of compound C45 (from the previous step; not more than 1.42 mmol) in dimethyl sulfoxide (15 ml) was added tetraethylammonium cyanide (708 mg; 4.53 mmol). The reaction mixture was heated at 80°C for 16 hours, after which it was cooled, poured into water and extracted with ethyl acetate (2×100 ml). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo. Chromatography on silica gel (gradient: 0% to 2.8% methanol in dichloromethane) gave the racemic mixture of compounds 1 and 2 as a yellowish foamy solid. Yield of racemic product: 349 mg; 0.984 mmol; 69% in two stages.

This substance was separated into its constituent enantiomers by supercritical fluid chromatography [column: Chiral Technologies Chiralpak AD, 5 μm; mobile phase: 7:3 carbon dioxide/(methanol containing 0.1% ammonium hydroxide)]. The first eluting enantiomer was referred to as compound 1 and the second eluting enantiomer as compound 2; both were obtained as solids. The indicated absolute configurations for compounds 1 and 2 were determined based on X-ray diffraction analysis performed on compound 2 (see below).

For compound 1, yield: 118 mg; 0.333 mmol; 34% as a result of separation. LCMS m/z 354.7 [M+H] ⁺ . ¹ H NMR (400 MHz, CD ₃ OD) δ 9.12 (s, 1H), 8.83-8.63 (vbr.m, 1H), 8.18 (d, J=9.0 Hz, 1H), 7.72 (dd, J =9.0, 2.0 Hz, 1H), 5.37-5.13 (vbr.m, 1H), 4.45-4.31 (m, 1H), 4.06-3.97 (m, 1H), 3.88 (ddd, J=12 .0, 12.0, 2.5 Hz, 1H), 3.21 (q, J=7.5 Hz, 2H), 2.94-2.44 (br.m, 2H), 2.88 (dd, half ABX pattern, J= 17.1, 4.5 Hz, 1H), 2.78 (breadth dd, half of the ABX pattern, J=17.1, 6.5 Hz, 1H), 2.31-2.14 (breadth m, 1H), 2.14-1.97 (breadth m, 1H), 1.52 (t, J=7.3 Hz, 3H).

For compound 2, yield: 88.8 mg; 0.250 mmol; 25% yield as a result of separation. LCMS m/z 354.7 [M+H] ⁺ . ¹ H NMR (400 MHz, CD ₃ OD) δ 9.11 (s, 1H), 8.82-8.59 (vbr.m, 1H), 8.17 (d, J=9.0 Hz, 1H), 7.71 (dd, J =9.0, 2.0 Hz, 1H), 5.39-5.12 (vbr.m, 1H), 4.44-4.31 (m, 1H), 4.06-3.96 (m, 1H), 3.88 (ddd, J=12 , 12.3 Hz, 1H), 3.20 (q, J=7.5 Hz, 2H), 2.88-2.69 (breadth m, 1H), 2.88 (dd, half ABX pattern, J=17.1, 4, 0 Hz, 1H), 2.78 (breadth dd, half of the ABX pattern, J=17.1, 6.5 Hz, 1H), 2.67-2.46 (breadth m, 1H), 2.29-2.14 (breadth m, 1H ), 2.14-1.97 (breadth m, 1H), 1.52 (t, J=7.3 Hz, 3H).

A sample of Compound 2 was crystallized from 2-methyltetrahydrofuran/hexanes by vapor diffusion and used to determine the absolute configuration by X-ray crystallography:

Single crystal X-ray diffraction analysis of compound 2

X-ray analysis of single crystals

Data collection was carried out on a Bruker APEX diffractometer at room temperature. Data collection consisted of omega and phi scans. The resolution was limited by crystal diffraction to approximately 0.9 angstroms.

The structure was determined by direct methods using the SHELX software package in the monoclinic space group P2 ₁ . The structure was then refined using the least squares method in the full matrix approximation. All non-hydrogen atoms were identified and refined using anisotropic displacement parameters.

Hydrogen atoms were placed in calculated positions and rotated about their carrier atoms. The final refinement included isotropic displacement parameters for all hydrogen atoms.

Absolute structure analysis using likelihood methods (Hooft, 2008) was performed using the PLATON program (Spek). The results show that the absolute structure is well defined. The method calculates that the probability that the structure is correct is 100.0. The Huft parameter is reported as 0.045 with an ESD of 0.002.

The final R-factor was 5.1%. The final difference Fourier map showed no gaps or shifts in the electron density.

Relevant crystal summaries, data acquisition, and refinement parameters are listed in Table F. Atomic coordinates, bond lengths, bond angles, and bias parameters are listed in Tables G, H, and J.

Software and bibliography

SHELXTL, Version 5.1, Bruker AXS, 1997.

PLATON, A.L. Spek, J. Appl. Cryst. 2003, 36, 7-13.

MERCURY, C.F. Macrae, P.R. Edington, P. McCabe, E. Pidcock, G.P. Shields, R. Taylor, M. Towler, and J. van de Streek, J. Appl. Cryst. 2006, 39, 453-457.

OLEX2, O.V. Dolomanov, L.J. Bourhis, R.J. Gildea, J. A. K. Howard, and H. Puschmann, J. Appl. Cryst. 2009, 42, 339-341.

R.W.W. Hooft, L.H. Straver, and A.L. Spek, J. Appl. Cryst. 2008, 41, 96-103.

H.D. Flack, Acta Cryst. 1983, A39, 867-881.

Examples 3 and 4

1-(4,4-Difluoro-1-methylpyrrolidin-3-yl)-2-[(4-methyl-2H-1,2,3-triazol-2-yl)methyl]-1H-imidazo[4,5 -c]quinolin-8-carbonitrile, ENT 1 (3), and 1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(4-methyl-2H-1,2,3- triazol-2-yl)methyl]-1H-imidazo[4,5-c]quinoline-8-carbonitrile, ENT 2 (4)

Stage 1. Synthesis of 4-bromo-5-methyl-1Н-1,2,3-triazole (С46).

N-Bromosuccinimide (5.89 g; 33.1 mmol) was added to a solution of 4-methyl-1H-1,2,3-triazole (2.50 g; 30.1 mmol) in chloroform (30 ml) and the reaction the mixture was stirred for 16 hours at room temperature (15°C). The mixture was then diluted with dichloromethane (100 ml), washed with water (2 x 100 ml), dried over sodium sulfate, filtered and concentrated in vacuo to give the product as a white solid (4.9 g) which was used directly in the next step.

Stage 2. Synthesis of tert-butyl-(4-bromo-5-methyl-2H-1,2,3-triazol-2-yl)acetate (С47).

tert-Butyl bromoacetate (8.8 g; 45 mmol) was added in one portion to a mixture of compound C46 (from the previous step; 4.9 g; not more than 30.1 mmol) and cesium carbonate (17.6 g; 54.0 mmol) in N,N-dimethylformamide (80 ml). The reaction mixture was stirred at room temperature (20°C) for 16 hours, after which it was diluted with water (100 ml) and was extracted with ethyl acetate (2×80 ml). The combined organic layers were washed with saturated aqueous sodium chloride (2×100 ml), dried over sodium sulfate, filtered and concentrated in vacuo. Chromatography on silica gel (gradient: 0% to 15% ethyl acetate in petroleum ether) gave the product as a colorless oil. Yield: 4.00 g; 14.5 mmol; 48% in 2 stages.

Stage 3. Synthesis of tert-butyl-(4-methyl-2Н-1,2,3-triazol-2-yl)acetate (С48), methyl-(4-methyl-2Н-1,2,3-triazol-2 -yl)acetate (C49) and (4-methyl-2H-1,2,3-triazol-2-yl)acetic acid (C50).

A mixture of compound C47 (3.50 g; 12.7 mmol) and palladium on carbon (10%; 500 mg) in methanol (35 ml) was stirred under hydrogen atmosphere (40 psi (275.8 kPa)) in for 4 hours at room temperature (17°C). The reaction mixture was filtered and the filtrate was concentrated in vacuo to give a yellow oil (3.00 g). Based on ¹ N NMR analysis, the product was determined to be a mixture of compound C48 (tert-butyl ether), compound C49 (methyl ester) and compound C50 (carboxylic acid); this substance was passed directly to the next stage for the hydrolysis of ester bonds. Peaks ¹ H NMR (400 MHz, CD ₃ OD) δ [7.50 (s) and 7.49 (s), total 1H], [5.23 (s), 5.17 (s) and 5.10 (s), total 2H], 3.75 ( s, from methyl ether), 2.30 (s, 3H), 1.46 (s, from tert-butyl ether).

Stage 4. Synthesis of (4-methyl-2H-1,2,3-triazol-2-yl)acetic acid (C50).

A mixture of compounds C48, C49 and C50 (from the previous step; 3.00 g; not more than 12.7 mmol) in trifluoroacetic acid (3 ml) was stirred for 2 hours at room temperature (17°C). After removing the solvent in vacuo, the residue was dissolved in tetrahydrofuran (10 ml) and treated with aqueous sodium hydroxide solution (2 M; 10 ml). The reaction mixture was stirred for 1 hour at room temperature (17° C.), concentrated in vacuo and partitioned between water (50 ml) and dichloromethane (20 ml). The aqueous layer was extracted with dichloromethane (2×20 ml) and then acidified with 1 M aqueous hydrochloric acid to pH 1. This acidic aqueous layer was extracted with ethyl acetate (3×40 ml) and the combined ethyl acetate layers were dried over sodium sulfate, filtered and concentrated under reduced pressure to give the product as a yellow solid. Yield: 1.9 g; 13 mmol; 100% for 2 stages. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.46 (s, 1H), 5.25 (s, 2H), 2.34 (s, 3H).

Step 5. Synthesis of N-{6-cyano-4-[(4,4-difluoro-1-methylpyrrolidin-3-yl)amino]quinolin-3-yl}-2-(4-methyl-2H-1,2 ,3-triazol-2-yl)acetamide (C51).

This experiment was carried out to obtain two identical batches. 1-[3-(Dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (139 mg; 0.725 mmol) was added to a solution of compound P4 (100 mg; 0.330 mmol) and compound C50 (55.8 mg; 0.395 mmol) in pyridine (1 .0 ml). The reaction mixture was then stirred at 25° C. for 1 hour, at the end of this time LCMS analysis showed conversion to the product: LCMS m/z 427.2 [M+H] ⁺ , the two batches were combined, diluted with water (50 ml) and was extracted with ethyl acetate (3×50 ml). The combined organic layers were concentrated in vacuo and purified by silica gel chromatography (gradient: 17% to 50% ethyl acetate in petroleum ether) to give the product as a white solid. Yield: 210 mg; 0.492 mmol; 75%. ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.84 (s, 1H), 8.29 (d, J=1.5 Hz, 1H), 8.11 (d, J=8.8 Hz, 1H), 8.08 (broad. s, 1H), 7.80 (dd, J=8.8, 1.5 Hz, 1H), 7.56 (s, 1H), 5.34 (s, 2H), 4.77 (broad d, J=10.8 Hz, 1H), 4.30-4.17 (m, 1H), 3.10 (dd, J=9.8, 6.4 Hz, 1H), 3.07-2.95 (m, 2H), 2.68 (ddd, J=9.8, 5 .9, 2.0 Hz, 1H), 2.42 (s, 3H), 2.40 (s, 3H).

Stage 6. Synthesis of 1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(4-methyl-2H-1,2,3-triazol-2-yl)methyl]-1H-imidazo [4,5-c]quinolin-8-carbonitrile, ENT 1 (3), and 1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(4-methyl-2H-1, 2,3-triazol-2-yl)methyl]-1H-imidazo[4,5-c]quinoline-8-carbonitrile, ENT 2 (4).

2,4,6-Tripropyl-1,3,5,2,4,6-trioxatrifosfinan 2,4,6-trioxide (50% solution in ethyl acetate; 0.92 ml; 1.5 mmol) was added to a solution of compound C51 (210 mg; 0.492 mmol) in N,N-dimethylformamide (1 ml) and propyl acetate (4 ml) at 15°C. The reaction mixture was stirred for 14 hours at 110°C, after which it was cooled and treated with aqueous sodium bicarbonate (60 ml). The resulting mixture was extracted with ethyl acetate (3 x 60 ml) and the combined organic layers concentrated in vacuo to give a racemic mixture of compounds 3 and 4 as a white solid. Yield of racemic product: 180 mg; 0.441 mmol; 90%.

This substance was separated into its constituent enantiomers by supercritical fluid chromatography [column: Regis Technologies, (S,S)-Whelk-0 ^® 1.10 μm; mobile phase: 55:45, carbon dioxide/(2-propanol containing 0.1% ammonium hydroxide)]. The product eluting first was designated Compound 3 and was obtained as a solid. Yield: 76.0 mg; 0.186 mmol; 42% as a result of separation. LCMS m/z 409.0 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 10.2-9.4 (vbr.s, 1H), 9.44 (s, 1H), 8.33 (d, J=8.8 Hz, 1H), 7.86 (dd, J= 8.6, 1.7 Hz, 1H), 7.43 (s, 1H), 6.41-6.09 (m, 2H), 5.96 (d, J=15.6 Hz, 1H), 3.75-3.57 (breadth m, 1H), 3.70 (dd, J=11.7, 11.7 Hz, 1H), 3.17-3.03 (m, 1H), 3.15 (dd, J=11.2, 11.2 Hz, 1H), 2.65 ( br.s, 3H), 2.32 (s, 3H).

The second eluted product, also isolated as a solid, was designated compound 4. Yield: 68.6 mg; 0.168 mmol; 38% as a result of separation. LCMS m/z 409.1 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 10.1-9.5 (vbr.s, 1H), 9.44 (s, 1H), 8.33 (d, J=8.8 Hz, 1H), 7.86 (dd, J= 8.8, 1.5 Hz, 1H), 7.43 (s, 1H), 6.36-6.10 (m, 2H), 5.96 (d, J=15.6 Hz, 1H), 3.75-3.57 (br. m, 1H), 3.70 (dd, J=11.7, 11.2 Hz, 1H), 3.17-3.03 (m, 1H), 3.15 (dd, J=11.7, 11.2 Hz, 1H), 2.65 ( br.s, 3H), 2.32 (s, 3H).

Example 5

Solution of compound P6 (75 mg; 0.24 mmol), (5-methyl-1,2-oxazol-3-yl) acetic acid (57.4 mg; 0.407 mmol) and N,N-diisopropylethylamine (0.11 ml ; 0.63 mmol) in tetrahydrofuran (4 ml) at 0°C was treated dropwise with 2,4,6-trioxide of 2,4,6-tripropyl-1,3,5,2,4,6-trioxatrifosfinan (50 % solution in ethyl acetate; 0.28 ml; 0.47 mmol) and the reaction mixture was left to warm to room temperature overnight. The resulting solution was concentrated under vacuum and the residue was dissolved in toluene (5 ml) and stirred at 110°C for 72 hours, then cooled to room temperature and partitioned between saturated aqueous sodium chloride and ethyl acetate. The organic layer was washed with saturated aqueous sodium chloride, dried over magnesium sulfate, filtered and concentrated. Chromatography on silica gel (gradient: 30% to 100% ethyl acetate in heptane) gave the product as a pale tan foam. Based on the analysis ^{of 1} H NMR, it was assumed that this substance existed as a mixture of rotamers. Yield: 79 mg; 0.189 mmol; 79%. LCMS m/z 419.5 (observed chlorine isotope pattern) [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ [9.27 (s) and 9.27 (s), total 1H], [8.52 (br.s) and 8.11 (br.s), total 1H], [8.22 (d, J=9.0 Hz) and 8.19 (d, J=9.0 Hz), total 1H], 7.66-7.57 (m, 1H), [6.11 (s) and 6.05 (s), total 1H], 5.69- 5.43 (m, 1H), [4.59 (AB quartet, J _AB = 16.8 Hz, Δν _Av = 19.5 Hz) and 4.50 (AB quartet, J _AB = 15.8 Hz, Δν _Av = 11.8 Hz ), total 2H], 4.43-4.27 (m, 2H), 3.92-3.63 (m, 2H), [3.30-3.17 (m) and 3.17-3.04 (m), total 1H], [2.40 (s) and 2.38 (s), total 3H], [2.23-2.14 (m) and 1.95-1.85 (m), total 1H].

Example 6

Step 1. Synthesis of lithium (6-methylpyrimidin-4-yl)acetate (С52).

n-Butyl lithium (2.5 M in hexanes; 5.00 ml; 12.5 mmol) was slowly added dropwise to a solution of 4,6-dimethylpyrimidine (1.08 g; 9.99 mmol) in tetrahydrofuran (20 ml) with temperature -78°С. After the reaction mixture was stirred for 20 minutes at -78°C, solid carbon dioxide (dry ice; 5.0 g) was added and the reaction mixture was warmed to room temperature (15°C) and stirred for 1 hour. Water (3.0 ml) was then added and the resulting mixture concentrated in vacuo to give the product as a white solid. Yield: 1.53 g; 9.68 mmol; 97%. ¹ H NMR (400 MHz, D ₂ O) δ 8.78 (s, 1H), 7.28 (s, 1H), [3.60 (s) and 3.59 (broad s), total 2H], 2.43 (s, 3H).

Step 2. Synthesis of 2-[(6-methylpyrimidin-4-yl)methyl]-1-[(3R)-1-methylpyrrolidin-3-yl]-1H-imidazo[4,5-c]quinoline-8 formate salt -carbonitrile (6).

This synthesis was carried out in a library format. A mixture of compound P9 (100 µmol), compound C52 (130 µmol) and 2,4,6-tripropyl-1,3,5,2,4,6-trioxatrifosfinan 2,4,6-trioxide (50% solution in ethyl acetate; 100 μl; 170 μmol) was treated with N,N-diisopropylethylamine (300 μmol) and 1,4-dioxane (1 ml) and the reaction vessel was sealed and shaken at 110° C. for 16 hours. After the solvents were removed using a ^Speedvac® concentrator, the residue was purified by reverse phase HPLC (column: Agela Durashell C18, 5 µm; mobile phase A: 0.225% formic acid in water; mobile phase B: acetonitrile; gradient: from 0% up to 31% B) to obtain the product. Yield: 1.5 mg; 3.5 µmol, 4%. LCMS m/z 384 [M+H] ⁺ . Retention time: 2.38 minutes (analytical HPLC conditions - column: Waters XBridge C18, 2.1×50 mm, 5 µm; mobile phase A: 0.05% ammonium hydroxide in water; mobile phase B: acetonitrile; gradient: 5 % B in 0.5 minutes; 5% to 100% B in 2.9 minutes; 100% B in 0.8 minutes; flow rate: 0.8 ml/minute).

Examples 7 and 8

8-Chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-[(5-methylpyrazin-2-yl)methyl]-1H-imidazo[4,5-c]quinoline, ENT 1 (7), and 8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-[(5-methylpyrazin-2-yl)methyl]-1H-imidazo[4.5 -c]quinoline, ENT 2 (8)

Stage 1. Synthesis of N-{6-chloro-4-[(3,3-difluorotetrahydro-2H-pyran-4-yl)amino]quinolin-3-yl}-2-(5-methylpyrazin-2-yl)acetamide (C53).

1-[3-(Dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (183 mg; 0.955 mmol) was added to a solution of compound P5 (150 mg; 0.478 mmol) and (5-methylpyrazin-2-yl)acetic acid (94.6 mg; 0.622 mmol) in pyridine (0.80 ml). The reaction mixture was stirred at 25°C for 4 hours, after which it was combined with a similar reaction mixture obtained using compound P5 (10.0 mg; 31.9 μmol), diluted with water (2 ml) and extracted with ethyl acetate (3× 3 ml). The combined organic layers were concentrated in vacuo to give the product as a brown oil which was used directly in the next step. Total yield: 214 mg; 0.478 mmol; 94%. LCMS m/z 448.2 [M+H] ⁺ .

Step 2. Synthesis of 8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-[(5-methylpyrazin-2-yl)methyl]-1H-imidazo[4,5-c ]quinoline, ENT 1 (7), and 8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-[(5-methylpyrazin-2-yl)methyl]-1H-imidazo [4,5-c]quinoline, ENT 2 (8).

2,4,6-Tripropyl-1,3,5,2,4,6-trioxatrifosfinan 2,4,6-trioxide (50% solution in ethyl acetate; 608 mg; 0.955 mmol) was added to a solution of compound C53 (214 mg; 0.478 mmol) in propyl acetate (1 ml) at 110°C, and the reaction mixture was stirred at 110°C for 48 hours. It was then concentrated in vacuo and purified by silica gel chromatography (gradient: 0% to 3% methanol in dichloromethane) to give a racemic mixture of compounds 7 and 8 as a yellow oil. Yield of racemic product: 150 mg; 0.349 mmol; 73%.

The enantiomers were separated using supercritical fluid chromatography ([column: Chiral Technologies ChiralCel OD, 5 µm; mobile phase: 7:3, carbon dioxide/(ethanol containing 0.1% ammonium hydroxide)]; then each enantiomer was separately purified by reverse phase HPLC (column: Agela Durashell, 5 µm; mobile phase A: 0.05% ammonium hydroxide in water; mobile phase B: acetonitrile; gradient: 32% to 52% B) The enantiomer eluting first was designated compound 7 , and the second eluting enantiomer as compound 8. Both compound 7 and compound 8 were obtained as solids, and based on analysis of ¹ H NMR spectra, it was suggested that both compounds exist as a mixture of rotamers.

For compound 7, yield: 21.3 mg; 49.6 µmol, 14% by separation. LCMS m/z 429.8 (observed chlorine isotope pattern) [M+H] ⁺ . ¹ H NMR (400 MHz, CD ₃ OD) δ [9.10 (s) and 9.06 (s), total 1H], 8.72-8.42 (m, 3H), [8.17 (d, J=8.8 Hz) and 8.15 (d, J=8.8 Hz), total 1H], 7.76-7.68 (m, 1H), [6.11-5.96 (m) and 5.96-5.80 (m), total 1H], 4.9-4.66 (m, 2H , estimated; partially overlain by water peak), 4.39-4.17 (m, 2H), 4.08-3.77 (m, 2H), [3.35-3.21 (m) and 3.17-3.04 (m), total 1H; supposed; partially covered by the solvent peak], [2.57 (s) and 2.54 (s), total 3H], [2.42-2.33 (m) and 2.32-2.21 (m), total 1H].

For compound 8, yield: 32.6 mg; 75.8 µmol, 22% by separation. LCMS m/z 429.7 (observed chlorine isotope pattern) [M+H] ⁺ . ¹ H NMR (400 MHz, CD ₃ OD) δ [9.10 (s) and 9.06 (s), total 1H], 8.71-8.43 (m, 3H), [8.17 (d, J=8.8 Hz) and 8.15 (d, J=9 Hz), total 1H], 7.76-7.69 (m, 1H), [6.10-5.96 (m) and 5.96-5.81 (m), total 1H], 4.9-4.67 (m, 2H, estimated ; partially overlain by a water peak), 4.39-4.17 (m, 2H), 4.08-3.77 (m, 2H), [3.35-3.21 (m) and 3.17-3.04 (m), total 1H; supposed; partially covered by the solvent peak], [2.57 (s) and 2.54 (s), total 3H], [2.42-2.33 (m) and 2.32-2.22 (m), total 1H].

Example 9

1-[(2R,4R)-2-Methyltetrahydro-2H-pyran-4-yl]-2-[(1-methyl-1H-1,2,3-triazol-4-yl)methyl]-8-( trifluoromethyl)-1H-imidazo[4,5-c]quinoline (9)

Stage 1. Synthesis of (1-methyl-1H-1,2,3-triazol-4-yl)methanol (С54).

Lithium aluminum hydride (685 mg; 18.0 mmol) was added to a suspension of ethyl 1-methyl-1H-1,2,3-triazole-4-carboxylate (1.40 g; 9.02 mmol) in tetrahydrofuran (20 ml) with a temperature of 0°C, and the reaction mixture was stirred at 0°C for 1 hour. Water was then added dropwise at 0° C. until gas evolution was observed, after which sodium sulfate was added and the mixture was stirred for 10 minutes. The mixture was then filtered and the filtrate concentrated in vacuo to give the product as a yellow oil. Yield: 700 mg; 6.19 mmol; 69%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 7.90 (s, 1H), 5.15 (t, J=5.5 Hz, 1H), 4.49 (d, J=5.5 Hz, 2H), 4.01 ( s, 3H).

Stage 2. Synthesis of (1-methyl-1H-1,2,3-triazol-4-yl)methylmethanesulfonate (C55).

Methanesulfonyl chloride (851 mg; 7.43 mmol) was added to a solution of compound C54 (700 mg; 6.19 mmol) and triethylamine (1.00 g; 9.88 mmol) in dichloromethane (20 ml) at 0°C. The reaction mixture was stirred at 0°C for 2 hours, after which water (100 ml) was added and the mixture was extracted with dichloromethane (2×100 ml). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo to give the product as a yellow oil which was used directly in the next step. Yield: 800 mg; 4.18 mmol; 68%.

Stage 3. Synthesis of (1-methyl-1H-1,2,3-triazol-4-yl)acetonitrile (C56).

To a solution of compound C55 (800 mg; 4.18 mmol) in acetonitrile (20 ml) was added potassium cyanide (1.50 g; 23.0 mmol). The reaction mixture was stirred at 60°C overnight, after which it was treated with water (150 ml) and was extracted with dichloromethane (3×100 ml). The combined organic layers were washed with saturated aqueous sodium chloride (80 ml), dried over sodium sulfate, filtered and concentrated in vacuo to give the product as a brown solid. Yield: 200 mg; 1.64 mmol; 39%. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.61 (s, 1H), 4.13 (s, 3H), 3.89 (br.s, 2H).

Stage 4. Synthesis of (1-methyl-1H-1,2,3-triazol-4-yl)acetic acid (C57).

A solution of compound C56 (200 mg; 1.64 mmol) in concentrated hydrochloric acid (4 ml) was stirred at 60° C. for 2 hours. After the reaction mixture was cooled to room temperature, it was diluted with water (10 ml) and washed with tert-butyl methyl ether (2×20 ml). The aqueous layer was then concentrated to dryness to give the product as a brown solid. Yield: 200 mg; 1.42 mmol; 87%. LCMS m/z 142.0 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 7.94 (s, 1H), 4.01 (s, 3H), 3.66 (s, 2H).

Step 5. Synthesis of N-[4-{[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]amino}-6-(trifluoromethyl)quinolin-3-yl]-2-(1-methyl -1H-1,2,3-triazol-4-yl)acetamide (C58).

1-[3-(Dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (118 mg; 0.615 mmol) was added in one portion to a solution of compound P7 (100 mg; 0.307 mmol) and compound C57 (52.1 mg; 0.369 mmol) in pyridine (0.8 ml), and the reaction mixture was stirred at 25°C for 16 hours. Then it was poured into water (50 ml) and extracted with ethyl acetate (3×30 ml); the combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo to give the product as a red oil (160 mg) which was used directly in the next step. LCMS m/z 449.2 [M+H] ⁺ .

Step 6. Synthesis of 1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-2-[(1-methyl-1H-1,2,3-triazol-4-yl)methyl] -8-(trifluoromethyl)-1H-imidazo[4,5-c]quinoline (9).

2,4,6-Tripropyl-1,3,5,2,4,6-trioxatrifosfinan 2,4,6-trioxide (1.6 M solution in ethyl acetate; 0.669 ml; 1.07 mmol) was added to a solution of compound C58 (from the previous step; not more than 0.307 mmol) in N,N-dimethylformamide (1 ml) and propyl acetate (4 ml). The reaction mixture was stirred at 110°C for 16 hours, after which it was poured into water (40 ml) and was extracted with ethyl acetate (2×30 ml). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo. Chromatography on silica gel (gradient: 0% to 1.5% methanol in dichloromethane) followed by reverse phase HPLC (column: Agela Durashell C18, 5 µm; mobile phase A: 0.05% ammonium hydroxide in water; mobile phase B : acetonitrile gradient: 5% to 95% B) gave the product as a solid. Yield: 29.5 mg; 68.5 µmol, 22% over two steps. LCMS m/z 431.1 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 9.35 (s, 1H), 9.13-8.89 (br. s, 1H), 8.38 (d, J=9.0 Hz, 1H), 7.86 (br. d, J =8.5 Hz, 1H), 7.64-7.54 (br.s, 1H), 5.53-5.38 (m, 1H), 4.62 (s, 2H), 4.29 (dd, J=12.0, 5.0 Hz , 1H), 4.07 (s, 3H), 3.83-3.68 (m, 2H), 2.77-2.57 (m, 1H), 2.50-2.31 (m, 1H), 2.0-1.59 (m, 2H, putative; partially overlapped water peak), 1.32 (d, J=6.5 Hz, 3H).

Examples 10 and 11

[cis-4-(8-Chloro-2-cyclobutyl-1H-imidazo[4,5-c]quinolin-1-yl)tetrahydro-2H-pyran-2-yl]acetonitrile, ENT 1 (10), and [ cis-4-(8-chloro-2-cyclobutyl-1H-imidazo[4,5-c]quinolin-1-yl)tetrahydro-2H-pyran-2-yl]acetonitrile, ENT 2 (11)

Step 1. Synthesis of N-[4-({cis-2-[(benzyloxy)methyl]tetrahydro-2H-pyran-4-yl}amino)-6-chloroquinolin-3-yl]cyclobutanecarboxamide (C59).

1-[3-(Dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (771 mg; 4.02 mmol) was added to a solution of compound P3 (800 mg; 2.01 mmol) and cyclobutanecarboxylic acid (221 mg; 2.21 mmol) in pyridine (20 ml). The reaction mixture was stirred at 25°C for 40 hours, after which it was concentrated under vacuum and partitioned between water (80 ml) and ethyl acetate (80 ml). The aqueous layer was extracted with ethyl acetate (80 ml) and the combined organic layers were dried over sodium sulfate, filtered and concentrated under reduced pressure to give the product as a foamy orange solid (1.01 g) which was used directly in the next step. LCMS m/z 479.9 (observed chlorine isotope pattern) [M+H] ⁺ .

Step 2. Synthesis of 1-{cis-2-[(benzyloxy)methyl]tetrahydro-2H-pyran-4-yl}-8-chloro-2-cyclobutyl-1H-imidazo[4,5-c]quinoline (С60) .

A solution of compound C59 (from the previous step; not more than 2.01 mmol) in acetic acid (20 ml) was stirred at 110° C. for 16 hours. This was combined with the same reaction mixture prepared using compound C59 (154 mg; 0.321 mmol) and concentrated in vacuo. The residue was mixed with half-saturated aqueous sodium bicarbonate (100 ml) and extracted with ethyl acetate (100 ml); the organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure to give the product as a yellow solid. Total yield: 1.07 g; 2.32 mmol; quantitative in two stages. LCMS m/z 462.0 (observed chlorine isotope pattern) [M+H] ⁺ .

Stage 3. Synthesis of [cis-4-(8-chloro-2-cyclobutyl-1H-imidazo[4,5-c]quinolin-1-yl)tetrahydro-2H-pyran-2-yl]methanol (C61).

Boron trichloride (1 M solution; 6.95 ml; 6.95 mmol) was added portionwise to a solution of compound C60 (1.07 g; 2.32 mmol) in dichloromethane (30 ml) at 10°C. The reaction mixture was stirred at 25°C for 1 hour, after which it was poured into saturated aqueous sodium bicarbonate (80 ml) and extracted with dichloromethane (2×50 ml). The combined organic layers were dried over sodium sulfate, filtered, concentrated in vacuo and purified using silica gel chromatography (gradient: 0% to 2% methanol in dichloromethane) to give the product as a yellowish solid. Yield: 643 mg; 1.73 mmol; 75%. LCMS m/z 371.9 (observed chlorine isotope pattern) [M+H] ⁺ .

Step 4. Synthesis of [cis-4-(8-chloro-2-cyclobutyl-1H-imidazo[4,5-c]quinolin-1-yl)tetrahydro-2H-pyran-2-yl]methylmethanesulfonate (C62).

Triethylamine (525 mg; 5.19 mmol) and methanesulfonyl chloride (0.160 ml; 2.07 mmol) were added to a solution of compound C61 (643 mg; 1.73 mmol) in dichloromethane (20 ml). The reaction mixture was stirred at 25°C for 1 hour, after which it was poured into water (50 ml) and was extracted with ethyl acetate (2×50 ml). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo to give the product as a light yellow foamy solid. Yield: 750 mg; 1.67 mmol; 96%. LCMS m/z 449.8 (observed chlorine isotope pattern) [M+H] ⁺ .

Step 5. Synthesis of [cis-4-(8-chloro-2-cyclobutyl-1H-imidazo[4,5-c]quinolin-1-yl)tetrahydro-2H-pyran-2-yl]acetonitrile, ENT 1 (10 ), and [cis-4-(8-chloro-2-cyclobutyl-1H-imidazo[4,5-c]quinolin-1-yl)tetrahydro-2H-pyran-2-yl]acetonitrile, ENT 2 (11) .

Tetraethylammonium cyanide (781 mg; 5.00 mmol) was added to a solution of compound C62 (750 mg; 1.67 mmol) in dimethyl sulfoxide (15 ml) and the reaction mixture was heated at 80° C. for 16 hours. Then it was diluted with tert-butyl methyl ether (100 ml) and washed successively with water (2×50 ml) and saturated aqueous sodium chloride solution (50 ml). The combined aqueous layers were extracted with tert-butyl methyl ether (50 ml) after which the combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo. Chromatography on silica gel (gradient: 0% to 2% methanol in dichloromethane) gave a racemic mixture of compounds 10 and 11 as a light yellow foamy solid. Yield of racemic product: 613 mg; 1.61 mmol; 96%.

A portion of this material (300 mg; 0.788 mmol) was separated into its constituent enantiomers by supercritical fluid chromatography [column: Chiral Technologies Chiralpak AS, 5 μm; mobile phase: 3:2, carbon dioxide/(ethanol containing 0.1% ammonium hydroxide)]. The enantiomer eluting first was designated compound 10 and was obtained as a solid. Yield: 91.1 mg; 0.239 mmol; 30% as a result of separation. LCMS m/z 381.0 (observed chlorine isotope pattern) [M+H] ⁺ . ¹ H NMR (400 MHz, CD ₃ OD) δ 9.14 (s, 1H), 8.72-8.55 (broad s, 1H), 8.17 (d, J=8.5 Hz, 1H), 7.71 (dd, J= 9.0, 2.0 Hz, 1H), 5.23-4.97 (vbr.m, 1H), 4.36 (dd, J=11.8, 5.3 Hz, 1H), 4.18-4.08 (m, 1H) , 4.03-3.95 (m, 1H), 3.86 (ddd, J=12.0, 12.0, 2.5 Hz, 1H), 2.88 (dd, half ABX pattern, J=17.1, 4.0 Hz , 1H), 2.77 (dd, half of the ABX pattern, J=17.1, 6.5 Hz, 1H), 2.73-2.42 (m, 6H), 2.33-1.93 (m, 4H).

The second enantiomer eluting, also isolated as a solid, was designated compound 11. Yield: 93.9 mg; 0.247 mmol; 31% as a result of separation. LCMS m/z 381.0 (observed chlorine isotope pattern) [M+H] ⁺ . ¹ H NMR (400 MHz, CD ₃ OD) δ 9.14 (s, 1H), 8.72-8.54 (broad s, 1H), 8.17 (d, J=9 Hz, 1H), 7.71 (d, J=9 Hz , 1H), 5.25-4.96 (vbr.m, 1H), 4.36 (dd, J=12.5 Hz, 1H), 4.19-4.07 (m, 1H), 4.03-3.95 (m, 1H), 3.86 ( broad.dd, J=12, 12Hz, 1H), 2.88 (dd, half ABX pattern, J=17.1, 4.0Hz, 1H), 2.77 (dd, half ABX pattern, J=17.1, 6.0 Hz, 1H), 2.73-2.42 (m, 6H), 2.33-1.92 (m, 4H).

Example 12

8-(Difluoromethyl)-2-[(4-methoxy-1H-pyrazol-1-yl)methyl]-1-[(2R,4R-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[ 4,5-s]quinoline (12)

Stage 1. Synthesis of ethyl-(4-methoxy-1H-pyrazol-1-yl)acetate (С63).

Ethyl bromoacetate (5.46 g; 32.7 mmol) was added in one portion to a mixture of 4-methoxy-1H-pyrazole hydrochloride (4.00 g; 29.7 mmol) and potassium carbonate (8.62 g; 62.4 mmol) in N,N-dimethylformamide (40 ml) at room temperature (30°C). The reaction mixture was stirred at room temperature (30°C) for 16 hours, after which it was diluted with water (200 ml) and was extracted with ethyl acetate (3×100 ml). The combined organic layers were washed with saturated aqueous sodium chloride (2×150 ml), dried over sodium sulfate, filtered and concentrated in vacuo. Chromatography on silica gel (gradient: 0% to 30% ethyl acetate in petroleum ether) gave the product as a colorless oil. Yield: 4.45 g; 24.2 mmol; 81%. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.30 (d, J=0.8 Hz, 1H), 7.15 (d, J=0.8 Hz, 1H), 4.80 (s, 2H), 4.24 (q, J=7.2 Hz, 2H), 3.76 (s, 3H), 1.29 (t, J=7.2 Hz, 3H).

Stage 2. Synthesis of (4-methoxy-1H-pyrazol-1-yl)acetic acid (C64).

An aqueous solution of sodium hydroxide (2 M; 24.2 ml; 48.4 mmol) was added in one portion to a solution of compound C63 (4.45 g; 24.2 mmol) in tetrahydrofuran (30 ml) at room temperature (29°C) , and the reaction mixture was stirred at room temperature (29°C) for 3 hours. It was then concentrated under reduced pressure, diluted with water (50 ml) and extracted with dichloromethane (2×30 ml). The organic layers were discarded and the aqueous layer was acidified to pH 1 with 1 M hydrochloric acid and extracted with ethyl acetate (4×50 ml). After the combined ethyl acetate layers were dried over sodium sulfate, they were filtered and concentrated in vacuo to give the product as a white solid. Yield: 2.80 g; 17.9 mmol; 74%. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 7.44 (s, 1H), 7.21 (s, 1H), 4.80 (s, 2H), 3.65 (s, 3H).

Stage 3. Synthesis of 8-(difluoromethyl)-2-[(4-methoxy-1H-pyrazol-1-yl)methyl]-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl] -1H-imidazo[4,5-c]quinoline (12).

To a solution of compound P11 (50 mg; 0.16 mmol) in toluene (1.5 ml) was added compound C64 (26.7 mg; 0.171 mmol) and N,N-diisopropylethylamine (31.2 μl; 0.179 mmol) followed by by adding 2,4,6-tripropyl-1,3,5,2,4,6-trioxatrifosfinan 2,4,6-trioxide (50% solution in ethyl acetate; 0.107 ml; 0.180 mmol). The reaction mixture was heated at 60°C for 90 minutes and then at 100°C for 4 hours, after which it was partitioned between ethyl acetate (10 ml) and saturated aqueous sodium bicarbonate (10 ml). The organic layer was dried over sodium sulfate, filtered, concentrated in vacuo and purified by silica gel chromatography (gradient: 0% to 15% methanol in dichloromethane) to give the product as an off-white solid. Yield: 51 mg; 0.12 mmol; 75%. LCMS m/z 428.5 [M+H] ⁺ . ¹ H NMR (400 MHz, CD ₃ OD) δ 9.28 (s, 1H), 8.98-8.81 (broad s, 1H), 8.34 (d, J=8.6 Hz, 1H), 7.92 (d, J= 9.0 Hz, 1H), 7.48 (s, 1H), 7.30 (s, 1H), 7.08 (t, J _HF =56.0 Hz, 1H), 5.82 (s, 2H), 5.44-5.29 (broad. m, 1H), 4.23 (dd, J=11.7, 5.1 Hz, 1H), 3.81-3.66 (m, 2H), 3.71 (s, 3H), 2.76-2.55 (breadth m, 1H), 2.47-2.24 (br. m, 1H), 1.90-1.56 (b. m, 2H), 1.28 (d, J=6.3 Hz, 3H).

Example 13

8-(Difluoromethyl)-2-[(5-methylpyrazin-2-yl)methyl]-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[4,5 -c]quinoline(13)

The reaction of compound P11 (50 mg; 0.16 mmol) with (5-methylpyrazin-2-yl)acetic acid was carried out using the method described for the synthesis of compound 12 from compound P11 in example 12. In this case, silica gel chromatography was performed twice ( gradient: 0% to 15% methanol in dichloromethane) to give the product as a light orange solid. Yield: 39 mg; 92 µmol, 58%. LCMS m/z 424.5 [M+H] ⁺ . ¹ H NMR (400 MHz, CD ₃ OD) δ 9.19 (s, 1H), 9.03-8.87 (broad s, 1H), 8.64 (s, 1H), 8.48 (s, 1H), 8.33 (d, J= 8.6 Hz, 1H), 7.90 (d, J=8.6 Hz, 1H), 7.08 (t, J _HF =56.0 Hz, 1H), 5.51-5.31 (breadth m, 1H), 4.80 ( s, 2H), 4.26 (dd, J=12.1, 5.1 Hz, 1H), 3.84-3.66 (m, 2H), 2.84-2.65 (br.m, 1H), 2.55 (s, 3H), 2.52-2.35 (br. m, 1H), 2.13-1.84 (b. m, 2H), 1.31 (d, J=5.9 Hz, 3H).

Examples 14 and 15

{8-Chloro-l-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[4,5-c]quinolin-2-yl}(5-methylpyrazin-2- yl)methanol, DIAST 1 (14), and {8-chloro-1-[(2R,4R)-2-methyl tetrahydro-2H-pyran-4-yl]-1H-imidazo[4,5-c]quinoline -2-yl}(5-methylpyrazin-2-yl)methanol, DIAST2 (15)

Stage 1. Synthesis of 8-chloro-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[4,5-c]quinoline (С65).

Formic acid (310 ml) was added to a mixture of iron powder (34.7 g; 621 mmol), ammonium chloride (33.2 g; 621 mmol) and compound C32 (20 g; 62.2 mmol) in 2-propanol (310 ml) at room temperature (14°C). The reaction mixture was heated at 80°C for 16 hours, after which it was diluted with ethanol (300 ml) and filtered. The collected solids were washed with 2-propanol (200 ml) and dichloromethane (100 ml) and the combined filtrates were concentrated in vacuo, then co-evaporated with ethanol (200 ml). The residue was diluted with dichloromethane (300 ml), basified by adding saturated aqueous sodium bicarbonate (500 ml) and then filtered through diatomaceous earth; the filter pack was washed with dichloromethane (300 ml). The aqueous layer of the combined filtrates was extracted with dichloromethane (4×100 ml) and the combined organic layers were washed with saturated aqueous sodium chloride (100 ml), dried over sodium sulfate, filtered and concentrated under reduced pressure. Chromatography on silica gel (gradient: 0% to 5% methanol in dichloromethane) gave a solid which was washed with a mixture of petroleum ether and ethyl acetate (3:1; 100 ml) and petroleum ether (50 ml) to give the product as a beige solid . Yield: 10.05 g; 33.3 mmol; 54%. LCMS m/z 301.8 (observed chlorine isotope pattern) [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 9.35 (s, 1H), 8.25 (d, J=9.0 Hz, 1H), 8.19 (s, 1H), 8.09 (d, J=2.3 Hz, 1H), 7.66 (dd, J=8.8, 2.3Hz, 1H), 5.02 (tt, J=12.0, 3.8Hz, 1H), 4.30 (ddd, J=11.9, 4 .6, 1.6 Hz, 1H), 3.77-3.89 (m, 2H), 2.33-2.46 (m, 2H), 2.09-2.22 (m, 1H), 1.83-1.95 (m, 1H), 1.38 (d , J=6.3 Hz, 3H).

Stage 2. Synthesis of {8-chloro-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[4,5-c]quinolin-2-yl}(5- methylpyrazin-2-yl)methanol, DIAST 1 (14), and {8-chloro-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[4,5- c]quinolin-2-yl}(5-methylpyrazin-2-yl)methanol, DIAST 2(15).

Compound C65 (100 mg; 0.33 mmol) was loaded into the tube and the tube was then evacuated and filled with nitrogen; this procedure was repeated twice, tetrahydrofuran (1.6 ml) was added and the solution was cooled to -78°C. A complex of 2,2,6,6-tetramethylpiperidinylmagnesium chloride and lithium chloride (1 M solution in tetrahydrofuran and toluene; 0.497 ml; 0.497 mmol) was added and the reaction mixture was left to stir for 1 hour at -78°C. In a separate tube, 5-methylpyrazine-2-carbaldehyde (80.9 mg; 0.662 mmol) was dissolved in tetrahydrofuran (1.6 ml) and the resulting solution was cooled in a dry ice-acetone bath for 10 minutes. This solution was then added to the reaction mixture, which was then left to stir while slowly heating to 15°C. After 1 hour, it was combined with two similar reaction mixtures obtained using compound C65 (50 mg; 0.17 mmol; 100 mg; 0.33 mmol) and the resulting mixture was diluted with water (20 ml) and extracted with ethyl acetate (3×15 ml). The combined organic layers were concentrated in vacuo and subjected to reverse phase HPLC (column: Phenomenex Synergi Max-RP, 10 µm; mobile phase A: 0.1% trifluoroacetic acid in water; mobile phase B: acetonitrile; gradient: 15% to 45 % C) to obtain a diastereomeric mixture of compounds 14 and 15 as a viscous brick-red oil. Combined yield of diastereomeric mixture: 180 mg; 0.425 mmol; 51%.

This substance was separated into its constituent diastereomers by supercritical fluid chromatography [column: Regis Technologies, (S,S)-Whelk-0 ^® 1, 10 μm; mobile phase: 3:2, carbon dioxide/(ethanol containing 0.1% ammonium hydroxide)]. The diastereomer eluting first, obtained as a light yellow solid, was designated compound 14. Yield: 58.6 mg; 0.138 mmol; 32% as a result of separation. LCMS m/z 423.9 (observed chlorine isotope pattern) [M+H] ⁺ . ¹ H NMR (400 MHz, CD ₃ OD) δ 9.12 (br.s, 1H), 8.92 (s, 1H), 8.83-8.74 (br.s, 1H), 8.48 (s, 1H), 8.18 (d, J=9.0Hz, 1H), 7.74(dd, J=9.0, 2.0Hz, 1H), 6.51(s, 1H), 5.58-5.46(m, 1H), 4.29(dd, J= 11.8, 5.3 Hz, 1H), 3.80-3.66 (br. m, 1H), 3.66-3.52 (b. m, 1H), 2.79-2.66 (m, 1H), 2.60 (s, 3H), 2.42-2.27 (br. m, 1H), 2.13-2.00 (br. m, 1H), 1.77-1.63 (br. m, 1H), 1.28 (br. d, J=5.5 Hz, 3H).

The second eluting diastereomer, also isolated as a light yellow solid, was designated compound 15. Yield: 56.8 mg; 0.134 mmol; 32% as a result of separation. LCMS m/z 423.9 (observed chlorine isotope pattern) [M+H] ⁺ . ¹ H NMR (400 MHz, CD ₃ OD) δ 9.12 (br.s, 1H), 8.92 (s, 1H), 8.82-8.74 (br.s, 1H), 8.47 (br.s, 1H), 8.18 ( d, J=8.5 Hz, 1H), 7.74 (dd, J=9.0, 2.0 Hz, 1H), 6.50 (s, 1H), 5.57-5.46 (m, 1H), 4.21 (dd, J=11.8, 4.8 Hz, 1H), 3.82-3.70 (br. m, 1H), 3.62-3.47 (b. m, 1H), 2.71-2.57 (b. m, 1H), 2.59 (s , 3H), 2.48-2.35 (m, 1H), 2.24-2.13 (br. m, 1H), 1.63-1.50 (b. m, 1H), 1.34 (d, J=6.0 Hz, 3H).

Examples 16 and 17

1-(4,4-Difluoro-1-methylpyrrolidin-3-yl)-8-fluoro-2-(1H-1,2,4-triazol-1-ylmethyl)-1H-imidazo[4,5-c] quinoline, ENT 1 (16), and 1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-8-fluoro-2-(1H-1,2,4-triazol-1-ylmethyl)-1H -imidazo[4,5-c]quinoline, ENT2 (17)

This interaction was carried out in a library format. N,N-Diisopropylethylamine (52 µl; 30 µmol) was added to a mixture of 1H-1,2,4-triazol-1-ylacetic acid (100 µmol) and compound P12 (29.6 mg; 100 µmol) in a mixture of ethyl acetate and toluene (0.5 ml), 3:2. 2,4,6-Tripropyl-1,3,5,2,4,6-trioxatrifosfinan 2,4,6-trioxide (50% solution in ethyl acetate; 0.19 mL; 0.32 mmol) was added and a reaction tube shaken and heated at 70°C for 10 hours, then at 110°C for 3 hours. The mixture was then partitioned between half-saturated aqueous sodium bicarbonate (1.5 ml) and ethyl acetate (2.4 ml) and subjected to vortex mixing. The organic layer was passed through a solid phase extraction cartridge (6 ml) filled with sodium sulfate (about 1 g); this extraction procedure was repeated twice and the combined eluates were concentrated in vacuo. Purification by reverse phase HPLC (column: Waters Sunfire C18, 5 µm; mobile phase A: 0.05% trifluoroacetic acid in water; mobile phase B: 0.05% trifluoroacetic acid in acetonitrile; gradient: 5% B per 1, 0 minute, then 5.0% to 75% B over 7.5 minutes, then 75% to 100% B) gave a racemic mixture of the two products. Separation into constituent enantiomers was performed using supercritical fluid chromatography [column: Chiral Technologies Chiralpak AD-H, 5 μm; mobile phase: 85:15 carbon dioxide/(methanol containing 0.2% ammonium hydroxide)]. The enantiomer eluting first was designated compound 16. Yield: 4.9 mg; 13 µmol, 13%. LCMS m/z 388.5 [M+H] ⁺ . Retention time: 2.91 minutes [analytical conditions, column: Chiral Technologies Chiralpak AD-H, 4.6×100 mm, 5 µm; mobile phase: 80:20 carbon dioxide/(methanol containing 0.2% ammonium hydroxide); back pressure: 150 bar (15 MPa); flow rate: 1.5 ml/minute].

The second enantiomer eluted was designated compound 17. Yield: 2.0 mg; 5.2 µmol, 5%. LCMS m/z 388.3 [M+H] ⁺ . Retention time: 3.31 minutes using the same analytical conditions.

Examples 18 and 19

1-(4,4-Difluoro-1-methylpyrrolidin-3-yl)-8-fluoro-2-[(4-methyl-1H-1,2,3-triazol-1-yl)methyl]-1H-imidazo [4,5-c]quinoline, ENT 1 (18), and 1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-8-fluoro-2-[(4-methyl-1H-1, 2,3-triazol-1-yl)methyl]-1H-imidazo[4,5-c]quinoline, ENT 2

(19)

(4-Methyl-1H-1,2,3-triazol-1-yl)acetic acid and compound P12 were used to form a racemic mixture of compounds 18 and 19 using the method described in examples 16 and 17. Separation into constituent enantiomers was carried out at using supercritical fluid chromatography [column: Chiral Technologies Chiralpak AD-H, 5 µm; mobile phase: 3:2, carbon dioxide/(methanol containing 0.2% ammonium hydroxide)]. 10 µmol, 10%. LCMS m/z 402.8 [M+H] ⁺ . Retention time: 1.68 minutes [analytical conditions, column: Chiral Technologies Chiralpak AD-H, 4.6×100 mm, 5 µm; mobile phase: 3:2 carbon dioxide/(methanol containing 0.2% ammonium hydroxide); back pressure: 150 bar (15 MPa); flow rate: 1.5 ml/minute].

The second enantiomer eluted was designated compound 19. Yield: 3.7 mg; 9.2 µmol, 9%. LCMS m/z 402.6 [M+H] ⁺ . Retention time: 4.1 minutes using the same analytical conditions.

Examples 20 and 21

1-(4,4-Difluoro-1-methylpyrrolidin-3-yl)-8-fluoro-2-[(5-methylpyrazin-2-yl)methyl]-1H-imidazo[4,5-c]quinoline, ENT 1 (20), and 1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-8-fluoro-2-[(5-methylpyrazin-2-yl)methyl]-1H-imidazo[4.5 -c]quinoline, ENT 2 (21)

(5-Methylpyrazin-2-yl)acetic acid and compound P12 were used to form a racemic mixture of compounds 20 and 21 using the method described in examples 16 and 17. Separation into its constituent enantiomers was carried out using supercritical fluid chromatography [column: Chiral Technologies Chiralpak AD-H, 5 µm; mobile phase: 85:15 carbon dioxide/(methanol containing 0.2% ammonium hydroxide)]. The enantiomer eluting first was designated compound 20. Yield: 2.0 mg; 4.8 µmol, 5%. LCMS m/z 413.9 [M+H] ⁺ . Retention time: 2.66 minutes [analytical conditions, column: Chiral Technologies Chiralpak AD-H, 4.6×100 mm, 5 µm; mobile phase: 80:20 carbon dioxide/(methanol containing 0.2% ammonium hydroxide); back pressure: 200 bar (20 MPa); flow rate: 1.5 ml/minute].

The second enantiomer eluted was designated compound 21. Yield: 1.8 mg; 4.4 µmol, 4%. LCMS m/z 413.9 [M+H] ⁺ . Retention time: 3.3 minutes using the same analytical conditions.

Examples 22 and 23

8-Chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-{[4-(methoxymethyl)-1H-1,2,3-triazol-1-yl]methyl}-1H -imidazo[4,5-c]quinoline, ENT 1 (22), and 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-{[4-(methoxymethyl)- 1H-1,2,3-triazol-1-yl]methyl}-1H-imidazo[4,5-c]quinoline, ENT 2 (23)

This interaction was carried out in a library format. N,N-Diisopropylethylamine (52 µl; 300 µmol) was added to a mixture of [4-(methoxymethyl)-1H-1,2,3-triazol-1-yl]acetic acid (which can be synthesized according to the method described by M.D Andrews et al., PCT International Patent Application WO 2014053967 A1, April 10, 2014; 100 µmol) and compound P13 (31.2 mg; 100 µmol) in a mixture of ethyl acetate and toluene (0.5 ml), 3:2 . Then 2,4,6-tripropyl-1,3,5,2,4,6-trioxatrifosfinan 2,4,6-trioxide (50% solution in ethyl acetate; 0.19 ml; 0.32 mmol) was added and the reaction the tube was shaken and heated at 70°C for 10 hours, then at 110°C for 3 hours. The reaction mixture was then partitioned between half-saturated aqueous sodium bicarbonate (1.5 ml) and ethyl acetate (2.4 ml) and subjected to vortex mixing. The organic layer was passed through a solid phase extraction cartridge (6 ml) filled with sodium sulfate (about 1 g); this extraction procedure was repeated twice and the combined eluates were concentrated in vacuo. Purification by reverse phase HPLC (column: Waters Sunfire C18, 5 µm; mobile phase A: 0.05% trifluoroacetic acid in water; mobile phase B: 0.05% trifluoroacetic acid in acetonitrile; gradient: 5% B over 1, 0 minute, then 5.0% to 75% B over 7.5 minutes, then 75% to 100% B) gave a racemic mixture of the two products. Separation into constituent enantiomers was performed using supercritical fluid chromatography [column: Chiral Technologies Chiralpak AD-H, 5 μm; mobile phase: 3:1, carbon dioxide/(methanol containing 0.2% ammonium hydroxide)]. The enantiomer eluting first was designated compound 22. Yield: 4.9 mg; 11 µmol, 11%. LCMS m/z 441.9 [M+H] ⁺ . Retention time: 2.4 minutes [analytical conditions, column: Chiral Technologies Chiralpak AD-H, 4.6×100 mm, 5 µm; mobile phase: 3:2 carbon dioxide/(methanol containing 0.2% ammonium hydroxide); back pressure: 150 bar (15 MPa); flow rate: 1.5 ml/minute].

The second enantiomer eluted was designated compound 23. Yield: 4.8 mg; 11 µmol, 11%. LCMS m/z 448.2 (observed chlorine isotope pattern) [M+H] ⁺ . Retention time: 2.95 minutes using the same analytical conditions.

Examples 24 and 25

8-Chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-(1Н-1,2,4-triazol-1-ylmethyl)-1Н-imidazo[4,5-с] quinoline, ENT 1 (24), and 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-(1H-1,2,4-triazol-1-ylmethyl)-1H -imidazo[4,5-c]quinoline, ENT2 (25)

1H-1,2,4-Triazol-1-ylacetic acid and compound P13 were used to form a racemic mixture of compounds 24 and 25 using the method described in examples 22 and 23. Separation into its constituent enantiomers was carried out using supercritical fluid chromatography [column: Chiral Technologies Chiralpak AD-H, 5 µm; mobile phase: 85:15 carbon dioxide/(methanol containing 0.2% ammonium hydroxide)]. In this case, the enantiomers were not completely separated, but the described samples were enriched in the indicated enantiomer. The enantiomer eluting first was designated compound 24. Yield: 2.3 mg; 5.7 µmol, 6%. LCMS m/z 404.5 (observed chlorine isotope pattern) [M+H] ⁺ . Retention time: 3.7 minutes [analytical conditions, column: Chiral Technologies Chiralpak AD-H, 4.6×100 mm, 5 µm; mobile phase: 75:25 carbon dioxide/(methanol containing 0.2% ammonium hydroxide); back pressure: 150 bar (15 MPa); flow rate: 1.5 ml/minute].

The second enantiomer eluted was designated compound 25. Yield: 1.0 mg; 2.5 µmol, 2%. LCMS m/z 403.9 [M+H] ⁺ . Retention time: 3.9 minutes using the same analytical conditions.

Examples 26 and 27

8-Chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-[(4-methoxy-1H-pyrazol-1-yl)methyl]-1H-imidazo[4,5-c ]quinoline, ENT 1 (26), and 8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-[(4-methoxy-1H-pyrazol-1-yl)methyl] -1H-imidazo[4,5-c]quinoline, ENT 2 (27)

This interaction was carried out in a library format.

N,N-Diisopropylethylamine (52 μl; 300 μmol) was added to a mixture of compound C64 (100 μmol) and compound P5 (31.2 mg; 99 μmol) in a mixture of ethyl acetate and toluene (0.5 ml) 3:2. Then 2,4,6-tripropyl-1,3,5,2,4,6-trioxatrifosfinan 2,4,6-trioxide (50% solution in ethyl acetate; 0.19 ml; 0.32 mmol) was added and the reaction the tube was shaken and heated at 70°C for 2 hours, then at 110°C for 6 hours. The reaction mixture was then partitioned between half-saturated aqueous sodium bicarbonate (1.5 ml) and ethyl acetate (2.4 ml) and subjected to vortex mixing. The organic layer was passed through a solid phase extraction cartridge (6 ml) filled with sodium sulfate (about 1 g); this extraction procedure was repeated twice and the combined eluates were concentrated in vacuo. Purification by reverse phase HPLC (column: Waters XBridge C18, 5 µm; mobile phase A: 0.03% ammonium hydroxide in water; mobile phase B: 0.03% ammonium hydroxide in acetonitrile; gradient: 5% to 100% C) gave a racemic mixture of the two products. Separation into constituent enantiomers was performed using supercritical fluid chromatography [column: Chiral Technologies Chiralcel OJ-H, 5 μm; mobile phase: 92:8, carbon dioxide/(methanol containing 0.2% ammonium hydroxide)]. The enantiomer eluting first was designated compound 26. Yield: 1.8 mg; 4.1 µmol, 4%. LCMS m/z 434.5 (observed chlorine isotope pattern) [M+H] ⁺ . Retention time: 1.98 minutes [analytical conditions, Chiral Technologies Chiralcel OJ-H, 4.6×100 mm, 5 µm; mobile phase: 90:10 carbon dioxide/(methanol containing 0.2% ammonium hydroxide); back pressure: 150 bar (15 MPa); flow rate: 1.5 ml/minute].

The second enantiomer eluted was designated compound 27. Yield: 1.8 mg; 4.1 µmol, 4%. LCMS m/z 435.5 [M+H] ⁺ . Retention time: 2.25 minutes using the same analytical conditions.

1. The required 6-fluoro-N ⁴ -[(2R,4R)-2 -methyltetrahydro-2H-pyran-4-yl]quinolin-3,4-diamine was synthesized from 6-fluoro-3-nitroquinolin-4-ol at using the general method described in the preparation of P7 for the synthesis of compound P7 from compound C25, except that the final reduction was carried out by hydrogenation over platinum on carbon, and not by treatment with iron powder and ammonium chloride.

2. Conditions for analytical HPLC. Column: Waters XBridge C18, 2.1×50 mm, 5 µm; mobile phase A: 0.0375% trifluoroacetic acid in water; mobile phase B: 0.01875% trifluoroacetic acid in acetonitrile; gradient: 1% to 5% B in 0.6 minutes; 5% to 100% B in 3.4 minutes; flow rate: 0.8 ml/minute.

3. In this case, the formation of amide and ring closure were carried out at separate stages: the condensation of the corresponding amine and carboxylic acid was carried out using 2,4,6-trioxide 2,4,6-tripropyl-1,3,5,2,4,6 -trioxatrifosfinan and triethylamine or N,N-diisopropylethylamine. The amide intermediate was cyclized by heating with 2,4,6-tripropyl-1,3,5,2,4,6-trioxatrifosfinan 2,4,6-trioxide and N,N-diisopropylethylamine in N,N-dimethylformamide.

4. Formation of an amide bond between compound P8 and cyclopentanecarboxylic acid was carried out using dimethyl carbonate and N,N-diisopropylethylamine to give N-(6-cyano-4-{[(2R,4R)-2-methyltetrahydro-2H-pyran-4- yl]amino}quinolin-3-yl)acetamide. This material was converted to Example 30 using the method described for the synthesis of C60 from C59 in Examples 10 and 11.

5. The racemate of Example 31 was separated into its constituent enantiomers by supercritical fluid chromatography [(column: Chiral Technologies Chiralpak AD, 5 µm; mobile phase: 3:1, carbon dioxide/(ethanol containing 0.1% ammonium hydroxide)] The compound eluted first was the compound of Example 31. The enantiomer of Example 31, [cis-4-(8-chloro-2-methyl-1H-imidazo[4,5-c]quinolin-1-yl)tetrahydro-2H- pyran-2-yl]acetonitrile, ENT 2, was the second enantiomer eluted, LCMS m/z 341.0 (observed chlorine isotope pattern) [M+H] ⁺ , and exhibited the following biological parameters: LRRK2, WT IC ₅₀ , 1660 nM.

6. The racemate of Example 32 was separated into its constituent enantiomers by supercritical fluid chromatography [column: Chiral Technologies Chiralpak AD, 5 µm; mobile phase: 7:3, carbon dioxide/(methanol containing 0.1% ammonium hydroxide)]. The compound eluted first was the compound of Example 32. The enantiomer of Example 32, 1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(5-methyl-1,2,4-oxadiazol-3 -yl)methyl]-1H-imidazo[4,5-c]quinoline-8-carbonitrile, ENT 2, was the second enantiomer eluted, LCMS m/z 409.8 [M+H] ⁺ , and exhibited the following biological parameters : LRRK2, WT IC ₅₀ , 473 nM.

7. Reaction of 5-methyl-1H-tetrazole with methyl bromoacetate in the presence of triethylamine gave methyl-(5-methyl-2H-tetrazol-2-yl)acetate, which was hydrolyzed with lithium hydroxide to give the desired (5-methyl-2H-tetrazol-2 -yl)acetic acid.

8. Conditions for analytical HPLC. Column: Waters XBridge C18, 2.1×50 mm, 5 µm; mobile phase A: 0.05% ammonium hydroxide in water; mobile phase B: acetonitrile; gradient: 5% B in 0.5 minutes; 5% to 100% B in 2.9 minutes; 100% B in 0.8 minutes; flow rate: 0.8 ml/minute.

9. Methyl-(5-methyl-1,3-oxazol-2-yl)acetate was synthesized using the method described by A.S.K. Hashmi et al., Organic Letters 2004, 6, 4391-4394. The hydrolysis of the ester was carried out using hydrochloric acid to give the desired (5-methyl-1,3-oxazol-2-yl)-acetic acid.

10. The desired 6-chloro-N ⁴ -[(3R)-1-methylpyrrolidin-3-yl]quinolin-3,4-diamine was synthesized from compound C7 using the method described in preparation of P9. The reduction of the nitro group in this case was carried out by hydrogenation over platinum(IV) oxide.

11. Conditions for analytical HPLC. Column: Waters Atlantis dC18, 4.6×50 mm, 5 µm; mobile phase A: 0.05% trifluoroacetic acid in water (v/v); mobile phase B: 0.05% trifluoroacetic acid in acetonitrile (v/v); gradient: 5.0% B over 1 minute, then linear from 5.0% to 95% B over 3.0 minutes, then 95% B over 1 minute; flow rate: 2 ml/minute.

12. The desired [4-(methoxymethyl)-1H-1,2,3-triazol-1-yl]acetic acid can be synthesized according to the method described by M. D. Andrews et al., PCT International Patent Application WO 2014053967 A1 , April 10, 2014.

13. The racemate of Example 46 was separated into its constituent enantiomers by supercritical fluid chromatography [column: Chiral Technologies ChiralCel OD, 5 µm; mobile phase: 7:3, carbon dioxide/(ethanol containing 0.1% ammonium hydroxide)]. The compound eluted first was the compound of Example 46. The enantiomer of Example 46, 8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-[(4-methyl-1H-1,2 ,3-triazol-1-yl)methyl]-1H-imidazo[4,5-c]quinoline, ENT 2, was the second enantiomer eluting, LCMS m/z 419.1 (observed chlorine isotope pattern) [M+H ] ⁺ , and showed the following biological parameters: LRRK2, WT IC ₅₀ , 21.4 nM; LRRK2, G2019S mutant IC ₅₀ , 16.1 nM.

14. The racemate of Example 47 was separated into its constituent enantiomers by supercritical fluid chromatography [column: Chiral Technologies Chiralpak AD-H, 5 µm; mobile phase: 65:35 carbon dioxide/(methanol containing 0.2% ammonium hydroxide)]. The second eluting compound was the compound of Example 47. The enantiomer of Example 47, 8-chloro-2-[(4-cyclopropyl-1H-1,2,3-triazol-1-yl)methyl]-1-(4,4 -difluoro-1-methylpyrrolidin-3-yl)-1H-imidazo[4,5-c]quinoline, ENT 1, was the first enantiomer eluted, LCMS m/z 444.3 [M+H] ⁺ , and exhibited the following biological parameters: LRRK2, WT IC ₅₀ , 97.3 nM.

15. Conditions for analytical HPLC. column: Chiral Technologies Chiralpak AD-H, 4.6×100 mm, 5 µm; mobile phase: 7:3 carbon dioxide/(methanol containing 0.2% ammonium hydroxide); back pressure: 200 bar (20 MPa); flow rate: 1.5 ml/minute.

16. The racemate of Example 48 (Example 82) was separated into its constituent enantiomers by supercritical fluid chromatography [column: Chiral Technologies Chiralpak AD-H, 5 µm; mobile phase: 4:1, carbon dioxide/(methanol containing 0.2% ammonium hydroxide)]. The second eluted compound was the compound of Example 48. The enantiomer of Example 48, 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(4-methyl-1H-1,2 ,3-triazol-1-yl)methyl]-1H-imidazo[4,5-c]quinoline, ENT 1, was the first enantiomer eluted, LCMS m/z 420.1 [M+H] ⁺ , and exhibited the following biological parameters: LRRK2, WT IC ₅₀ , 145 nM.

17. Conditions for analytical HPLC. Column: Chiral Technologies Chiralpak AD-H, 4.6×100 mm, 5 µm; mobile phase: 3:2 carbon dioxide/(methanol containing 0.2% ammonium hydroxide); back pressure: 120 bar (12 MPa); flow rate: 1.5 ml/minute.

18. The racemate of Example 49 was separated into its constituent enantiomers by supercritical fluid chromatography [column: Chiral Technologies Chiralpak AD-H, 5 µm; mobile phase: 3:1, carbon dioxide/(methanol containing 0.2% ammonium hydroxide)]. The compound eluted first was the compound of Example 49. The enantiomer of Example 49, 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(5-methyl-2/7-tetrazole -2-yl)methyl]-1H-imidazo[4,5-c]quinoline, ENT 2, was the second enantiomer eluted, LCMS m/z 421.1 [M+H] ⁺ , and exhibited the following biological parameters: LRRK2 , WT IC ₅₀ , 46.2 nM; LRRK2.

19. Conditions for analytical HPLC. Column: Chiral Technologies Chiralpak AD-H, 4.6×100 mm, 5 µm; mobile phase: 1:1 carbon dioxide/(methanol containing 0.2% ammonium hydroxide); back pressure: 120 bar (12 MPa); flow rate: 1.5 ml/minute.

20. The racemate of Example 50 was separated into its constituent enantiomers by supercritical fluid chromatography [column: Chiral Technologies Chiralpak AD-H, 5 µm; mobile phase: 3:1, carbon dioxide/(methanol containing 0.2% ammonium hydroxide)]. The compound eluted first was the compound of Example 50. The enantiomer of Example 50, 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(5-methylpyrazin-2-yl)methyl ]-1H-imidazo[4,5-c]quinoline, ENT 2, was the second enantiomer eluted, LCMS m/z 429.2 (observed chlorine isotope pattern) [M+H] ⁺ , and exhibited the following biological parameters: LRRK2 , WT IC ₅₀ , 181 nM.

21. The racemate of Examples 51 and 52 was separated into its constituent enantiomers by supercritical fluid chromatography [column: Chiral Technologies Chiralpak AD-H, 5 µm; mobile phase: 3:1, carbon dioxide/(methanol containing 0.2% ammonium hydroxide)]. The first eluting compound was Example 51 and the second eluting enantiomer was Example 52.

22. The racemate of Example 53 was separated into its constituent enantiomers by supercritical fluid chromatography [column: Chiral Technologies Chiralpak AD-H, 5 µm; mobile phase: 55:45 carbon dioxide/(methanol containing 0.2% ammonium hydroxide)]. The second eluting compound was the compound of Example 53. The enantiomer of Example 53, 1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-8-fluoro-2-{[4-(methoxymethyl)-1H-1 ,2,3-triazol-1-yl]methyl}-1H-imidazo[4,5-c]quinoline, ENT 1, was the first enantiomer eluted, LCMS m/z 432.7 [M+H] ⁺ , and showed the following biological parameters: LRRK2, WT IC ₅₀ , 229 nM.

23. The racemate of Example 54 was separated into its constituent enantiomers by supercritical fluid chromatography [Chiral Technologies Chiralpak AD-H, 5 μm; mobile phase: 4:1, carbon dioxide/(methanol containing 0.2% ammonium hydroxide)]. The second eluting compound was the compound of Example 54. The enantiomer of Example 54, 8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-(1H-1,2,4-triazol- 1-ylmethyl)-1H-imidazo[4,5-c]quinoline, ENT 1 (Example 93), was the first enantiomer eluted, LCMS m/z 405.3 [M+H] ⁺ , and exhibited the following biological parameters: LRRK2, WT IC ₅₀ , 11.0 nM; LRRK2.

24. Conditions for analytical HPLC. Column: Phenomenex Lux Amylose-1, 4.6×100 mm, 5 µm; mobile phase: 7:3 carbon dioxide/(methanol containing 0.2% ammonium hydroxide); back pressure: 150 bar (15 MPa); flow rate: 1.5 ml/minute.

25. The racemate of Example 55 was separated into its constituent enantiomers by supercritical fluid chromatography [column: Chiral Technologies ChiralCel OD-H, 5 µm; mobile phase: 7:3, carbon dioxide/(methanol containing 0.2% ammonium hydroxide)]. The compound eluted first was the compound of Example 55. The enantiomer of Example 55, 8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-{[4-(methoxymethyl)-1H-1 ,2,3-triazol-1-yl]methyl}-1H-imidazo[4,5-c]quinoline, ENT 2, was the second enantiomer eluting, LCMS m/z 449.3 (observed chlorine isotope pattern) [M +H] ⁺ and showed the following biological parameters: LRRK2, WT IC ₅₀ , 15.3 nM.

26. Conditions of analytical HPLC. Column: Chiral Technologies ChiralCel OD-H, 4.6×100 mm, 5 µm; mobile phase: 3:2 carbon dioxide/(methanol containing 0.2% ammonium hydroxide); back pressure: 120 bar (12 MPa); flow rate: 1.5 ml/minute.

1. tert-Butyl-(3R)-3-aminopyrrolidin-1-carboxylate and compound C11 were used to synthesize tert-butyl-(3R)-3-[(3-amino-6-cyanoquinolin-4-yl)amino]pyrrolidine -1-carboxylate according to the method described for the synthesis of compound P9 in the preparation of P9. This substance was converted to tert-butyl-(3R)-3-{8-cyano-2-[(5-methyl-1,2,4-oxadiazol-3-yl)methyl]-1H-imidazo[4,5- c]quinolin-1-yl}pyrrolidine-1-carboxylate using the method described for the synthesis of compounds 3 and 4 in examples 3 and 4. Deprotection with trifluoroacetic acid was followed by alkylation with 2,2,2-trifluoroethyltrifluoromethanesulfonate and N,N-diisopropylethylamine to give the compound of Example 56.

2. Conditions for analytical HPLC. Column: Waters XBridge C18, 2.1×50 mm, 5 µm; mobile phase A: 0.0375% trifluoroacetic acid in water; mobile phase B: 0.01875% trifluoroacetic acid in acetonitrile; gradient: 1% to 5% B in 0.6 minutes; 5% to 100% B in 3.4 minutes; flow rate: 0.8 ml/minute.

3. Reaction of compound P3 with (5-methyl-1,2-oxazol-3-yl)acetic acid, 2,4,6-trioxide 2,4,6-tripropyl-1,3,5,2,4,6 -trioxatrifosfinan and N,N-diisopropylethylamine gave 1-{cis-2-[(benzyloxy)methyl]tetrahydro-2H-pyran-4-yl}-8-chloro-2-[(5-methyl-1,2-oxazole -3-yl)methyl]-1H-imidazo[4,5-c]quinoline, which was debenzylated with boron trichloride and oxidized using Dess-Martin periodinan [1,1,1-tris(acetyloxy)-1,1-dihydro- 1,2-benziodoxol-3-(1H)-one]. The resulting cis-4-{8-chloro-2-[(5-methyl-1,2-oxazol-3-yl)methyl]-1H-imidazo[4,5-c]quinolin-1-yl}tetrahydro-2H -pyran-2-carbaldehyde was converted into racemic 8-chloro-1-[cis-2-(difluoromethyl)tetrahydro-2H-pyran-4-yl]-2-[(5-methyl-1,2-oxazol-3- yl)methyl]-1H-imidazo[4,5-c]quinoline by treatment with (diethylamino)sulfur trifluoride.

4. The compound of Example 59 was isolated from the corresponding racemic mixture by supercritical fluid chromatography (column: Chiral Technologies Chiralpak AD, 5 μm; mobile phase: 7:3, carbon dioxide/methanol). Example 59 was the first enantiomer eluted. The enantiomer of Example 59, 8-chloro-1-[cis-2-(difluoromethyl)tetrahydro-2H-pyran-4-yl]-2-[(5-methyl-1,2-oxazol-3-yl)methyl] -1H-imidazo[4,5-c]quinoline, ENT 2, was the second enantiomer eluted, LCMS m/z 433.0 [M+H] ⁺ , and exhibited the following biological parameters: LRRK2, WT IC ₅₀ , 631 nM .

5. The compound of Example 60 was isolated from the corresponding racemic mixture by supercritical fluid chromatography [column: Chiral Technologies Chiralcel OJ-H, 5 μm; mobile phase: 85:15, carbon dioxide/(ethanol containing 0.2% ammonium hydroxide)]. Example 60 was the eluting first enantiomer. The enantiomer of Example 60, 8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-[(5-methyl-1,2-oxazol-3-yl)methyl]-1H- imidazo[4,5-c]quinoline, ENT 2, was the second enantiomer eluted, LCMS m/z 419.3 [M+H] ⁺ , and exhibited the following biological parameters: LRRK2, WT IC ₅₀ , 42.2 nM.

6. Reaction of 1,2,3-thiadiazol-4-ylmethanol with methanesulfonyl chloride and triethylamine followed by displacement using potassium cyanide and hydrolysis in concentrated hydrochloric acid gave the desired 1,2,3-thiadiazol-4-ylacetic acid.

7. Conditions for analytical HPLC. Column: Waters XBridge C18, 2.1×50 mm, 5 µm; mobile phase A: 0.0375% trifluoroacetic acid in water; mobile phase B: 0.01875% trifluoroacetic acid in acetonitrile; gradient: 10% to 100% B in 4.0 minutes; flow rate: 0.8 ml/minute.

8. The compound of Example 75 was isolated from the corresponding racemic mixture by supercritical fluid chromatography [column: Chiral Technologies Chiralpak IC, 10 μm; mobile phase: 55:45 carbon dioxide/(2-propanol containing 0.1% ammonium hydroxide)]. Example 75 was the eluting first enantiomer. The enantiomer of Example 75, 8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-[(5-methyl-1,2,4-oxadiazol-3-yl)methyl]- 1H-imidazo[4,5-c]quinoline, ENT 2, was the second enantiomer eluted, LCMS m/z 420.0 (observed chlorine isotope pattern) [M+H] ⁺ , and exhibited the following biological parameters: LRRK2, WT IC ₅₀ not determined.

9. Treatment of ethyl 3-oxobutanoate with lithium trifluoromethanesulfonate, trifluoromethanesulfonic anhydride, and N,N-diisopropylethylamine gave ethyl 3-{[(trifluoromethyl)sulfonyl]oxy}but-2-enoate. This was reacted with zinc cyanide in the presence of tetrakis(triphenylphosphine)palladium(0) to give ethyl 3-cyanobut-2-enoate, which was hydrogenated over palladium on carbon followed by hydrolysis with sodium hydroxide to give the desired 3-cyanobutanoic acid.

10. The compound of Example 78 was isolated from the corresponding diastereomeric mixture by supercritical fluid chromatography [column: Phenomenex Lux Cellulose-2, 10 μm; mobile phase: 3:2, carbon dioxide/(2-propanol containing 0.1% ammonium hydroxide)]. Example 78 was the second eluting diastereomer. The diastereomer of Example 78, 3-{8-chloro-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[4,5-c]quinolin-2-yl} -2-methylpropanenitrile, DIAST 1, was the first eluting diastereomer, LCMS m/z 369.0 (observed chlorine isotope pattern) [M+H] ⁺ , and exhibited the following biological parameters: LRRK2, WT IC ₅₀ , 37.2 nM .

11. The compound of Example 79 was isolated from the corresponding racemic mixture by supercritical fluid chromatography [column: Chiral Technologies Chiralpak AY, 10 μm; mobile phase: 3:2, carbon dioxide/(ethanol containing 0.1% ammonium hydroxide)]. Example 79 was the eluting second enantiomer. The enantiomer of Example 79, 8-fluoro-1-[cis-3-fluorocyclopentyl]-2-(1,2,3-thiadiazol-4-ylmethyl)-1H-imidazo[4,5-c]quinoline, ENT 1, was the first enantiomer eluted, LCMS m/z 372.0 [M+H] ⁺ , and exhibited the following biological parameters: LRRK2, WT IC ₅₀ , 7.54 nM.

12. Reaction of 5-methyl-1H-tetrazole with methyl bromoacetate in the presence of triethylamine gave methyl-(5-methyl-2H-tetrazol-2-yl)acetate, which was hydrolyzed with lithium hydroxide to give the desired (5-methyl-2H-tetrazol-2 -yl)acetic acid.

13. Conditions for analytical HPLC. Column: Waters Atlantis dC18, 4.6×50 mm, 5 µm; mobile phase A: 0.05% trifluoroacetic acid in water (v/v); mobile phase B: 0.05% trifluoroacetic acid in acetonitrile (v/v); gradient: 5.0% B over 1 minute, then linear from 5.0% to 95% B over 3.0 minutes, then 95% B over 1 minute. Flow rate: 2 ml/minute.

14. Conditions for analytical HPLC. Column: Chiral Technologies Chiralpak AD-H, 4.6×100 mm, 5 µm; mobile phase: 75:25 carbon dioxide/(methanol containing 0.2% ammonium hydroxide); back pressure: 200 bar (20 MPa); flow rate: 1.5 ml/minute.

15. The compound of Example 85 was isolated from the corresponding racemic mixture by supercritical fluid chromatography [column: Chiral Technologies Chiralpak AD-H, 5 μm; mobile phase: 4:1 carbon dioxide/(methanol containing 0.2% ammonium hydroxide)]. Example 85 was the first enantiomer eluted. The enantiomer of Example 85, 1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-8-fluoro-2-[(2-methylimidazo[2,1-b][1,3,4]thiadiazol- 6-yl)methyl]-1H-imidazo[4,5-c]quinoline, ENT 2, was the second enantiomer eluted, LCMS m/z 458.3 [M+H] ⁺ , and exhibited the following biological parameters: LRRK2, WT IC ₅₀ , 55.9 nM.

16. Conditions for analytical HPLC. Column: Chiral Technologies Chiralpak AD-H, 4.6×100 mm, 5 µm; mobile phase: 70:30 carbon dioxide/(methanol containing 0.2% ammonium hydroxide); back pressure: 150 bar (15 MPa); flow rate: 1.5 ml/minute.

17. The compound of Example 83 was separated into its constituent enantiomers by supercritical fluid chromatography [column: Chiral Technologies Chiralpak AD-H, 5 µm; mobile phase: 4:1 carbon dioxide/(methanol containing 0.2% ammonium hydroxide)]. Example 86 was the eluting second enantiomer. The enantiomer of Example 86, 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-(1H-tetrazol-1-ylmethyl)-1H-imidazo[4,5-c]quinoline , ENT 1, was the first enantiomer eluted, LCMS m/z 407.1 [M+H] ⁺ , and exhibited the following biological parameters: LRRK2, WT IC ₅₀ , 271 nM.

18. Conditions of analytical HPLC. Column: Chiral Technologies Chiralcel OD-H, 4.6×100 mm, 5 µm; mobile phase: 7:3 carbon dioxide/(methanol containing 0.2% ammonium hydroxide); back pressure: 150 bar (15 MPa); flow rate: 1.5 ml/minute.

19. Example 87 was isolated from the corresponding racemic mixture by supercritical fluid chromatography [column: Chiral Technologies ChiralCel OD-H, 5 µm; mobile phase: 4:1 carbon dioxide/(methanol containing 0.2% ammonium hydroxide)]. Example 87 was the first enantiomer eluted. The enantiomer of example 87, 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(4-methyl-2H-1,2,3-triazol-2-yl)methyl ]-1H-imidazo[4,5-c]quinoline, ENT 2, was the second enantiomer eluted, LCMS m/z 420.2 [M+H] ⁺ , and exhibited the following biological parameters: LRRK2, WT IC ₅₀ , 37 .8 nM.

20. Analytical HPLC conditions. Column: Chiral Technologies Chiralpak AD-H, 4.6×100 mm, 5 µm; mobile phase: 60:40 carbon dioxide/(methanol containing 0.2% ammonium hydroxide); back pressure: 120 bar (12 MPa); flow rate: 1.5 ml/minute.

21. Example 88 was isolated from the corresponding racemic mixture by supercritical fluid chromatography [column: Chiral Technologies Chiralpak AD-H, 5 µm; mobile phase: 3:2, carbon dioxide/(methanol containing 0.2% ammonium hydroxide)]. Example 88 was the first enantiomer eluted. The enantiomer of example 88, 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(5-methyl-1,2,4-oxadiazol-3-yl)methyl]- 1H-imidazo[4,5-c]quinoline, ENT 2, was the second enantiomer eluted, LCMS m/z 420.5 [M+H] ⁺ , and exhibited the following biological parameters: LRRK2, WT IC ₅₀ , 261 nM.

22. Conditions of analytical HPLC. Column: Chiral Technologies Chiralpak AD-H, 4.6×100 mm, 5 µm; mobile phase: 7:3 carbon dioxide/(methanol containing 0.2% ammonium hydroxide); back pressure: 200 bar (20 MPa); flow rate: 1.5 ml/minute.

23. Example 89 was isolated from the corresponding racemic mixture by supercritical fluid chromatography [column: Chiral Technologies Chiralpak AD-H, 5 µm; mobile phase: 85:15 carbon dioxide/(methanol containing 0.2% ammonium hydroxide)]. Example 89 was the eluting second enantiomer. The enantiomer of example 89, 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(5-methyl-1,3,4-thiadiazol-2-yl)methyl]- 1H-imidazo[4,5-c]quinoline, ENT 1, was the first enantiomer eluted, LCMS m/z 434.8 [M+H] ⁺ , and exhibited the following biological parameters: LRRK2, WT IC ₅₀ , not determined.

24. Conditions for analytical HPLC. Column: Chiral Technologies Chiralcel OJ-H, 4.6×100 mm, 5 µm; mobile phase: 9:1 carbon dioxide/(methanol containing 0.2% ammonium hydroxide); back pressure: 150 bar (15 MPa); flow rate: 1.5 ml/minute.

25. Example 92 was isolated from the corresponding racemic mixture by supercritical fluid chromatography [column: Chiral Technologies Chiralcel OJ-H, 5 µm; mobile phase: 95:5, carbon dioxide/(methanol containing 0.2% ammonium hydroxide)]. Example 92 was the first enantiomer eluted. The enantiomer of example 92, 8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-[(5-methyl-1,3,4-thiadiazol-2-yl)methyl]- 1H-imidazo[4,5-c]quinoline, ENT 2, was the second enantiomer eluted, LCMS m/z 436.5 (observed chlorine isotope pattern) [M+H] ⁺ , and exhibited the following biological parameters: LRRK2, WT IC ₅₀ , 33.7 nM.

26. Conditions of analytical HPLC. Column: Phenomenex Lux Amylose-1, 4.6×100 mm, 5 µm; mobile phase: 7:3 carbon dioxide/(methanol containing 0.2% ammonium hydroxide); back pressure: 150 bar (15 MPa); flow rate: 1.5 ml/minute.

27. The compound of Example 93 was isolated from the corresponding racemic mixture by supercritical fluid chromatography [column: Chiral Technologies Chiralpak AD-H, 5 μm; mobile phase: 4:1 carbon dioxide/(methanol containing 0.2% ammonium hydroxide)]. Example 93 was the first enantiomer eluted. The enantiomer of Example 93, 8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-(1H-1,2,4-triazol-1-ylmethyl)-1H-imidazo[4 ,5-c]quinoline, ENT 2, was the second enantiomer eluted, LCMS m/z 405.6 [M+H] ⁺ , and showed the following biological parameters: LRRK2, WT IC ₅₀ , 10.3 nM.

28. Example 94 was isolated from the corresponding racemic mixture by supercritical fluid chromatography [column: Chiral Technologies Chiralpak AD-H, 5 µm; mobile phase: 3:1, carbon dioxide/(methanol containing 0.2% ammonium hydroxide)]. Example 94 was the first enantiomer eluted. The enantiomer of Example 94, 8-chloro-2-[(4-cyclopropyl-1H-1,2,3-triazol-1-yl)methyl]-1-(3,3-difluorotetrahydro-2H-pyran-4-yl )-1H-imidazo[4,5-c]quinoline, ENT 2, was the second enantiomer eluted, LCMS m/z 445.3 (observed chlorine isotope pattern) [M+H] ⁺ , and exhibited the following biological parameters: LRRK2 , WT IC ₅₀ , 9.35 nM.

Example 95

[5-({8-Chloro-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[4,5-c]quinolin-2-yl}methyl)pyrazine -2-yl]methanol (95)

Stage 1. Synthesis of benzyl-2-(5-methylpyrazin-2-yl)acetate

Suspension containing 2-(5-methylpyrazin-2-yl)acetic acid (1.00 g; 6.57 mmol) and benzyl alcohol (853 mg; 7.89 mmol; 0.820 ml) in tetrahydrofuran (26.3 ml) treated with N,N-diisopropylethylamine (1.72 ml; 9.86 mmol) and 2,4,6-tripropyl-1,3,5,2,4,6-trioxatrifosfinan 2,4,6-trioxide (50% solution in N,N-dimethylformamide, 4.69 ml, 7.89 mmol). The solids slowly dissolved when the reaction mixture was stirred at room temperature for 20 hours. The reaction mixture was quenched with saturated aqueous sodium bicarbonate and then extracted with ethyl acetate. The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo. Chromatography on silica gel (gradient: 0% to 80% ethyl acetate in heptane) gave the product as a yellow oil. Yield: 1.2 g; 76%. LCMS m/z 243.4 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.48 (d, J=1.5 Hz, 1H), 8.43 (d, J=1.5 Hz, 1H), 7.43-7.30 (m, 5H), 5.20 ( s, 2Н), 3.91 (s, 2Н), 2.58 (s, 3Н).

Stage 2. Synthesis of benzyl-(5-methyl-4-oxidopyrazin-2-yl)acetate

A solution of benzyl-2-(5-methylpyrazin-2-yl)acetate (1.22 g; 5.03 mmol) in dichloromethane (50 ml) was placed under local vacuum line and the reaction flask was refilled with nitrogen; this procedure was repeated three times. The solution was cooled to 0°C and meta-chloroperbenzoic acid (mCPBA; 886 mg; 5.13 mmol) was added in one portion keeping the temperature of the solution at 0°C. The reaction mixture was allowed to slowly warm to room temperature and stirred for 20 hours, after which it was quenched with saturated aqueous sodium bicarbonate. The aqueous layer was extracted with dichloromethane and the combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (gradient: 0% to 80% ethyl acetate in heptane) to give the product as a colorless oil which turned into a white solid on standing. A 2D NMR NOE (Nuclear Overhauser Effect) spectroscopy study showed that this substance was the desired regioisomer. Yield: 616 mg; 47%. LCMS m/z 259.2 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.41 (s, 1H), 8.20 (s, 1H), 7.44-7.31 (m, 5H), 5.20 (s, 2H), 3.84 (s, 2H), 2.47 ( s, 3H).

A regioisomeric N-oxide (200 mg; 15%) was also isolated, as well as some starting material (205 mg; 17%).

Stage 3. Synthesis of benzyl-{5-[(acetyloxy)methyl]pyrazin-2-yl}acetate

A solution of benzyl-(5-methyl-4-oxidopyrazin-2-yl)acetate (591 mg; 2.29 mmol) in acetic anhydride (9.15 ml) was heated to 70°C for 1 hour, and then at 100 °C within 24 hours. The reaction mixture was then cooled to room temperature, and acetic anhydride and acetic acid were removed under vacuum on a rotary evaporator. The residue was dissolved in ethyl acetate and washed with saturated aqueous sodium bicarbonate. The organic layer was dried over sodium sulfate, filtered and concentrated in vacuo. Chromatography on silica gel (gradient: 0% to 70% ethyl acetate in heptane) gave the product as a yellow oil. Yield: 392 mg; 57%. LCMS m/z 301.2 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.62 (d, J=1.5 Hz, 1H), 8.59 (d, J=1.5 Hz, 1H), 7.44-7.31 (m, 5H), 5.27 ( s, 2Н), 5.20 (s, 2Н), 3.96 (s, 2Н), 2.19 (s, 3Н).

Step 4. Synthesis of {5-[(acetyloxy)methyl]pyrazin-2-yl}acetic acid

A mixture of benzyl-{5-[(acetyloxy)methyl]pyrazin-2-yl}acetate (390 mg; 1.30 mmol) and palladium on carbon (150 mg; 10% Pd on a dry basis) in ethyl acetate (13, 0 ml) was placed in a Hastelloy reactor and purged with nitrogen three times, and then purged with hydrogen three times. The reaction mixture was stirred at room temperature under a hydrogen pressure of 30 psi (206.8 kPa) for 2 hours, after which it was filtered. The filter cake was washed with ethyl acetate and the combined filtrates were concentrated in vacuo to give the product as a yellow oil. Yield: 186 mg; product yield 68%. Analysis of the spectral data and thin layer chromatography showed that the product was contaminated with the hydrogenation product of the acetoxy group (approximately 3:4 methyl relative to acetoxymethyl, as determined by NMR). This substance was passed to the next stage without further purification.

Step 5. Synthesis of [5-({8-chloro-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[4,5-c]quinolin-2-yl }methyl)pyrazin-2-yl]methyl acetate

Mixture of compound P10 (246 mg; 0.843 mmol) and {5-[(acetyloxy)methyl]pyrazin-2-yl}acetic acid (186 mg; 0.885 mmol; as a mixture from the previous step) in toluene (17.7 ml) treated with N,N-diisopropylethylamine (176 μl; 1.01 mmol) and 2,4,6-tripropyl-1,3,5,2,4,6-trioxatrifosfinan 2,4,6-trioxide (50% solution in ethyl acetate ; 1.51 ml; 2.53 mmol). The reaction mixture was heated to 70°C for 1 hour and then at 110°C for 4 hours. The reaction mixture was allowed to cool to ambient temperature and then quenched by adding saturated aqueous sodium bicarbonate and extracted with ethyl acetate. The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo. Chromatography on silica gel (gradient: 0% to 10% methanol in dichloromethane) gave two products. The desired product was obtained as a light brown oil. Yield: 206 mg; fifty%. LCMS m/z 466.2 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 9.28 (s, 1H), 8.73 (s, 1H), 8.67 (s, 1H), 8.59 (s, 1H), 8.24 (d, J=9.0 Hz, 1H), 7.66 (dd, J=8.9, 2.1 Hz, 1H), 5.30 (broad s, 1H), 5.27 (s, 2H), 4.73 (s, 2H), 4.34 (dd, J= 12.0, 5.1 Hz, 1H), 3.73 (broad s, 2H), 2.75 (broad s, 1H), 2.48 (broad s, 1H), 2.17 (s, 3H), 1.85 (b.s. s, 1H), 1.74 (br.s, 1H), 1.38 (d, J=6.1 Hz, 3H). A light yellow solid was also obtained, identified as the deacetoxy product 8-chloro-2-[(5-methylpyrazin-2-yl)methyl]-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4- yl]-1H-imidazo[4,5-c]quinoline (95A). Yield: 131 mg; 36%.

Step 6. Synthesis of [5-({8-chloro-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[4,5-c]quinolin-2-yl }methyl)pyrazin-2-yl]methanol (95)

In a solution of [5-({8-chloro-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[4,5-c]quinolin-2-yl}methyl )pyrazin-2-yl]methyl acetate (206 mg; 0.442 mmol) in methanol (10 mL) was added potassium carbonate (61.1 mg; 0.442 mmol). The resulting white suspension was stirred at room temperature for 30 minutes, after which it was diluted with water and extracted with dichloromethane. The organic layer was dried over sodium sulfate, filtered and concentrated in vacuo. Chromatography on silica gel (gradient: 0% to 20% methanol in dichloromethane) gave a light yellow foam (151 mg). This material was recrystallized from diethyl ether and heptane to give the product as a light yellow solid. Yield: 130 mg; 69%. LCMS m/z 424.2 [M+H] ⁺ . ¹ H NMR (400 MHz, CDCl ₃ ) δ 9.29 (s, 1H), 8.72-8.67 (m, 2H), 8.59 (s, 1H), 8.24 (d, J=8.9 Hz, 1H), 7.66 ( dd, J=8.9, 2.1 Hz, 1H), 5.31 (br.s, 1H), 4.87 (d, J=5.4 Hz, 2H), 4.73 (s, 2H), 4.34 (dd, J=12.1, 5.2 Hz, 1H), 3.74 (br.s, 2H), 2.91 (br.s, 1H), 2.76 (br.s, 1H), 2.48 (br.s, 1H), 1.88 (br s, 1H), 1.75 (br s, 1H), 1.38 (d, J=6.1 Hz, 3H).

Example 96

8-Chloro-2-{[5-( ² H ₃ )methylpyrazin-2-yl]methyl}-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[ 4,5-c]quinoline (96); [5-({8-chloro-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[4,5-c]quinolin-2-yl}methyl)pyrazine -2-yl]( ² H ₂ ) methanol (96B)

To 1.2 g of 8-chloro-2-[(5-methylpyrazin-2-yl)methyl]-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[ 4,5-c]quinoline (95A) (yellow solid) was added 5.7 g of deuterated acetic acid (CD ₃ CO ₂ D) in the first container. The mixture was stirred at 120°C for 20 hours and then concentrated. Proton NMR analysis showed over 90% D/H exchange in the pyrazine methyl group.

In the second container to 3.0 g of 8-chloro-2-[(5-methylpyrazin-2-yl)methyl]-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H -imidazo[4,5-c]quinolone was added 50 ml of deuterated acetic acid. The mixture was stirred at 120°C for 24 hours and then concentrated.

The concentrated residues from the first and second containers were combined and dissolved in 75 ml of deuterated acetic acid. This solution was stirred at 120°C for 24 hours and then concentrated. The residue was dissolved in 50 ml of deuterated acetic acid and stirred at 120°C for 24 hours and then concentrated. The residue was dissolved in 120 ml of ethyl acetate and washed with 60 ml of a saturated aqueous sodium carbonate solution. The organic layer was dried over magnesium sulfate and concentrated to give 4.4 g of a dark solid.

A portion of this sample, 2.4 g, was dissolved in 100 ml of acetic acid and stirred at room temperature for 24 hours and then concentrated. The residue was dissolved in 100 ml of acetic acid and stirred at room temperature for 24 hours and concentrated. The residue was dissolved in 150 ml of ethyl acetate, washed with 80 ml of a mixture of brine/ammonium hydroxide, 3:1. The organic layer was dried over magnesium sulfate, concentrated to give 2.4 g of a dark substance, which was purified using the step gradient method (hold 20% B 0 to 1.5 minutes, 20% to 70% B 1.5 to 10 minutes and finally 70 to 100% B 10 to 12 minutes; with mobile phase A: 0.05% formic acid in water and mobile phase B: 0.05% formic acid in acetonitrile) on a Phenomenex Gemini NX C18 150 mm column × 21.2 mm 5, µm with a flow rate of 27 ml/min. The collected fractions were lyophilized to give samples of a yellowish fibrous solid with a total weight of 2.12 g.

Analytical data: [M+H] ⁺ determined 411.178 (calculated 411.178); retention time at HPLC 4.12 min on column C ¹⁸ 100 mm × 3.0 mm, 2.6 µm, with 5% B 0 to 1.5 min, 5 to 100% B 1.5 to 4.0 min and hold at 100% for 4.0 to 5.4 minutes (A: 0.1% formic acid in water, B: 0.1% formic acid in acetonitrile); ¹ H NMR (600 MHz, DMSO-d6) δ 9.17 (s, 1H), 8.66 (s, 2H), 8.46 (d, J=1.5 Hz, 1H), 8.19 (d, J=8.9 Hz , 1H), 7.74 (dd, J=8.9, 2.2 Hz, 1H), 5.27 (m, 1H), 4.77 (s, 2H), 4.16 (m, 1H), 3.69 (m, 1H), 3.60 (m, 1H), 2.47 (m, 1H), 2.21 (m, 1H), 2.09-1.92 (m, 1H), 1.85 (m, 1H), 1.22 (d, J=6.1 Hz, 3H) .

Biological analyzes

LRRK2 analysis

LRRK2 kinase activity was measured using Lantha Screen technology from Invitrogen. GST-labeled truncated LRRK2 from Invitrogen (p/n PV4874) was incubated with a fluorescein labeled ezrin/radixin/moesin complex (ERM) peptide substrate, also known as LRRKtide (Invitrogen p/n PR8976A), in the presence of a dose of the compound. At the end of the assay, the assay was stopped and detection was performed with a terbium labeled antiphospho-ERM antibody (Invitrogen cat. no. PR8975A). The analysis was carried out according to the following protocol. An effective compound dose was prepared by diluting the compound to an upper concentration of 0.3 mM in 100% DMSO and making serial semi-log dilutions in DMSO to give an 11-point curve, 100-fold of the final analyte concentration. Using an Echo sonic dosing system, 60 nL of compounds were transferred to a 384-well Corning small volume assay plate. 3 µl substrate working solution (200 nM LRRKtide, 2 mM ATP (adenosine triphosphate)), prepared in assay buffer (50 mM HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid), pH 7.5, 3 mM MgCl ₂ with 2 mM DTT (dithiothreitol) and 0.01% Brij35 added fresh) was added to the assay plate 60 nl of the compound. The kinase reaction was initiated with 3 μl of the LRRK2 enzyme working solution at a concentration of 4 μg/ml. Final reaction concentrations were 100 nM LRRKtide, 1 mM ATP, 2 μg/ml of LRRK2 enzyme, and a 3 μM upper dose compound dose. The reaction was allowed to proceed at room temperature for 30 minutes and then stopped by adding 6 μl of detection buffer (20 mM Tris (tris (hydroxymethyl) aminomethane) pH 7.6, 0.01% NP-40, 6 mM EDTA (ethylenediaminetetraacetic acid) with 2 nM terbium labeled antiphospho-ERM). After incubation for 1 hour at room temperature, the plate was read on an Envision analyzer with an excitation wavelength of 340 nm and detection of radiation at both 520 nm and 495 nm. The ratio of emission at 520 nm and 495 nm was used for data analysis. Inhibition of the G2019S mutant LRRK2 (Invitrogen cat. no. PV4881) was measured in exactly the same way. All final concentrations of substrate ATP and enzyme were the same.

a. Undefined

b. In this case, biological data was obtained using the formate salt of the compound of Example.

Intrinsic clearance (CL _int ) in human liver microsomes

Incubation mixtures (in duplicate) contained either Compound 95A or Compound 96 or Compound 95A and Compound 96 at final concentrations of 1 μM, human liver microsomes (BD Biosciences Bedford, MA, 0.25 μM CYP protein (cytochrome P450), which equivalent to a protein concentration of 0.801 mg/ml), NADPH (1.3 mM), MgCl ₂ (3.3 mM) and potassium phosphate buffer (100 mM; pH 7.4). The final volume of the reaction mixture (500 μl) contained 0.003% DMSO, 0.5% acetonitrile. Incubations were carried out at 37°C, aliquots were taken (50 µl) at time points 0, 5, 10, 15, 20, 30, 45 and 60 minutes and quenched by adding to cold acetonitrile containing an internal standard (200 µl) of mass spectrometry (MS). The quenched incubations were vortexed for 1 minute followed by centrifugation at 3000 rpm for 5 minutes at room temperature (Allegra X-12R, Beckman Coulter, Fullerton, CA). The supernatant (150 µl) was then withdrawn and added to a 96-well deep-well sample injection plate containing 150 µl of water with 0.1% formic acid (v/v), the plates were capped and vortexed for 1 minute and then analyzed using LC-MS/MS as described below. Control incubation mixtures were prepared similarly without the addition of the NADPH cofactor to control any metabolism without the involvement of CYP/FMO (flavin-containing monooxygenase). Individual calibration curves (0.5-2000 nM) were generated, processed and analyzed as described above.

The amount of substrate (compounds 95A or 96) and metabolite (compounds 95 or 96B) was measured and the results are presented in Table 4a and Table 4b. The compound of Example 96 has a reduced internal clearance (increased half-life - T _1/2 ) compared to its corresponding non-deuterated form (compound 95A), which may be beneficial (eg, reduced dosage) while providing useful properties. In addition, the compound of example 96 has a lower metabolite formation rate (compound 96B) compared to the non-deuterated metabolite (compound 95) formed from the compound of example 95A. In the pooled substrates (competition) experiment, the compound of Example 96 shows a reduced intrinsic clearance (increased T _1/2 ) compared to its corresponding non-deuterated form (compound 95A) and has a lower rate of metabolite formation compared to the non-deuterated form (compound 95A) .

Intrinsic clearance (CL _int ) in cynomolgus monkey liver microsomes

Incubation mixtures (in duplicate) contained either Compound 95A or Compound 96 or Compound 95A and Compound 96 at 1 μM final concentrations, pooled cynomolgus liver microsomes (Xenotech, LLC, Lenexa, KS, 0.25 μM CYP protein, equivalent to protein concentration 0.21 mg/ml), NADPH (1.3 mM), magnesium chloride (3.3 mM), and potassium phosphate buffer (100 mM; pH 7.4). The final volume of the reaction mixture (500 μl) contained 0.003% DMSO, 0.5% acetonitrile. Incubations were carried out at 37°C and aliquots (50 µl) were taken at time points 0, 5, 10, 15, 20, 30, 45 and 60 minutes and quenched by adding to cold acetonitrile containing an internal standard (200 µl) wt. spectrometry (MS). The quenched incubations were vortexed for 1 minute followed by centrifugation at 3000 rpm for 5 minutes at room temperature (Allegra X-12R, Beckman Coulter, Fullerton, CA). The supernatant (150 μl) was then withdrawn and added to a 96-well deep well sample injection plate containing 150 μl of water with 0.1% formic acid (v/v). The plates were capped and vortexed for 1 minute and then analyzed using LC-MS/MS as described below. Control incubation mixtures were prepared similarly without the addition of the NADPH cofactor to control any metabolism without the involvement of CYP/FMO (flavin-containing monooxygenase). Individual calibration curves (0.5-2000 nM) were generated, processed and analyzed as described above.

The amount of substrate (compounds 95A or 96) and metabolite (compounds 95 or 96B) was measured and the results are presented in Table 5a and Table 5b. The compound of example 96 has a reduced intrinsic clearance (increased half-life - T _1/2 ) compared to its corresponding non-deuterated form (compound 95A), which may be beneficial (eg, reduced dosage) while providing useful properties. In addition, the compound of example 96 has a lower rate of metabolite formation (compound 96B) compared to the non-deuterated form (compound 95A). In the pooled substrates (competition) experiment, the compound of Example 96 showed a similar trend when compared to incubation with single substrates. The compound of example 96 showed a reduced intrinsic clearance (increased half-life - T _1/2 ) compared to its corresponding non-deuterated form (compound 95A) and has a lower rate of metabolite formation compared to the non-deuterated form (compound 95A).

Enzymatic kinetics in human liver microsomes

Incubation mixtures (in triplicate) contained compound 95A or compound 96 (1-1000 μM, final concentrations), pooled human liver microsomes (BD Biosciences, Bedford, MA, 0.25 protein concentration), NADPH (1.3 mM), magnesium chloride (5 mM); and potassium phosphate buffer (100 mM; pH 7.4). The final volume of the reaction mixture (100 μl) contained 1% acetonitrile. Incubations were carried out at 37°C. At 15 minutes for Compound 95A or 30 minutes for Compound 96, 50 μl of incubation fluid was quenched by adding 200 μl of cold acetonitrile containing 0.1% formic acid (v/v) and an internal mass spectrometry standard (MS). Quenched samples were vortexed for 1 minute followed by centrifugation at 3000 rpm for 5 minutes at room temperature (Allegra X-12R, Beckman Coulter, Fullerton, CA). The supernatant (150 μl) was placed in a clean sample injection block and dried completely under nitrogen atmosphere, then redissolved in 150 μl of water containing 0.1% formic acid (v/v). The plates were capped and vortexed for 1 minute and then analyzed using LC-MS/MS as described below. Metabolite production 95 (from Substrate 95A) and 96B (from Substrate 96) was quantified using a calibration curve (0.5-2000 nM) prepared using a Synthetic Substrate Standard 95. Standard curve samples were prepared, processed and analyzed as described above. .

The 95 or 96B metabolite formation kinetics as measured in human liver microsomes is shown in Table 6. Similar to the above in this example, the compound of Example 96 has a reduced intrinsic clearance compared to its corresponding non-deuterated form (compound 95A), which may be beneficial (e.g. , reduced dosage) while providing beneficial properties.

LC-MS/MS analyzes of the data presented in tables 4a, 4b, 5a, 5b and 6

The disappearance of 95A and 96 substrates and the formation of 95 or 96 B metabolites was determined using an LC-MS/MS system, which consists of an AB Sciex 6500 triple quadrupole mass spectrometer equipped with an electrospray ion source (AB Sciex, Framingham, MA) and an Agilent liquid chromatograph. Technologies Infinity 1290 (Santa Clara, CA). A binary gradient was used with a flow rate of 0.500 ml/min using 0.1% formic acid in water as the aqueous mobile phase (solvent A) and 0.1% formic acid in acetonitrile (solvent B) as the organic phase. The LC profile of the gradient starts with 5% solvent B, which was brought to 98% B in 2 minutes and then held for 0.20 minutes and returned to initial conditions (5% B) in 0.5 minutes, with a total chromatogram registration time of 3 .00 minutes. The analytical column used was Phenomenex Kinetex 2.6 µm, 2.1×50 mm (Phenomenex, Torrance, CA) with an injection volume of 10 µl. The mass spectrometer was launched in the mode of recording positive ions with the source temperature set to 500°C and the ionization voltage value of 4.5 kV. The following MS/MS transitions were used: for substrate 95A (408→310), substrate 96 (411→313), metabolite 95 (424→326), and metabolite 96B (426→328). Analytes were quantified using Analyst software version 1.6.2 or earlier (AB Sciex, Framingham, MA).

In this application, references are made to various publications. The disclosures of these publications are incorporated by reference in this application in their entirety for all purposes.

Those skilled in the art will appreciate that various modifications and variations may be made to the present invention without departing from the scope or spirit of the invention. Other embodiments of the invention will be apparent to those skilled in the art from a consideration of the description and practice of the invention disclosed herein. It is intended that the description of the invention and the examples be taken as examples only, and the true scope and spirit of the invention is indicated in the appended claims.

Claims (176)

1. Compound of formula I

where R ¹ is selected from the group consisting of methyl, ethyl, cyclobutyl, cyclopentyl,

R ² is selected from the group consisting of 2,2-difluoropropyl,

and R ³ is selected from the group consisting of fluorine, chlorine, cyano, difluoromethyl and trifluoromethyl; or a pharmaceutically acceptable salt thereof, wherein the compound or pharmaceutically acceptable salt has one or more hydrogen atoms which is replaced by a deuterium atom, provided that one or more hydrogen atoms are not replaced by only a deuterium atom in one or more of the 2 positions, 5, 7 or 8 quinoline ring. 2. A compound selected from the group consisting of: [(2S,4R)-4-(8-chloro-2-ethyl-1H-imidazo[4,5-c]quinolin-1-yl)tetrahydro-2H-pyran-2-yl]acetonitrile; [(2R,4S)-4-(8-chloro-2-ethyl-1H-imidazo[4,5-c]quinolin-1-yl)tetrahydro-2H-pyran-2-yl]acetonitrile; 1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(4-methyl-2H-1,2,3-triazol-2-yl)methyl]-1H-imidazo[4,5 -c]quinoline-8-carbonitrile, ENT 1 (enantiomer 1); 1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(4-methyl-2H-1,2,3-triazol-2-yl)methyl]-1H-imidazo[4,5 -c]quinoline-8-carbonitrile, ENT 2 (enantiomer 2); 8-chloro-1-[(4S)-3,3-difluorotetrahydro-2H-pyran-4-yl]-2-[(5-methyl-1,2-oxazol-3-yl)methyl]-1H-imidazo [4,5-c]quinoline; 2-[(6-methylpyrimidin-4-yl)methyl]-1-[(3R)-1-methylpyrrolidin-3-yl]-1H-imidazo[4,5-c]quinolin-8-carbonitrile; 8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-[(5-methylpyrazin-2-yl)methyl]-1H-imidazo[4,5-c]quinoline, ENT 1 (enantiomer 1); 8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-[(5-methylpyrazin-2-yl)methyl]-1H-imidazo[4,5-c]quinoline, ENT 2 (enantiomer 2); 1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-2-[(1-methyl-1H-1,2,3-triazol-4-yl)methyl]-8-( trifluoromethyl)-1H-imidazo[4,5-c]quinoline; [cis-4-(8-chloro-2-cyclobutyl-1H-imidazo[4,5-c]quinolin-1-yl)tetrahydro-2H-pyran-2-yl]acetonitrile, ENT 1 (enantiomer 1); [cis-4-(8-chloro-2-cyclobutyl-1H-imidazo[4,5-c]quinolin-1-yl)tetrahydro-2H-pyran-2-yl]acetonitrile, ENT 2 (enantiomer 2); 8-(difluoromethyl)-2-[(4-methoxy-1H-pyrazol-1-yl)methyl]-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo [4,5-c]quinoline; 8-(difluoromethyl)-2-[(5-methylpyrazin-2-yl)methyl]-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[4,5 -c]quinoline; {8-chloro-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[4,5-c]quinolin-2-yl}(5-methylpyrazin-2- yl) methanol, DIAST 1 (diastereomer 1); {8-chloro-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[4,5-c]quinolin-2-yl}(5-methylpyrazin-2- yl) methanol, DIAST 2 (diastereomer 2); 1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-8-fluoro-2-(1H-1,2,4-triazol-1-ylmethyl)-1H-imidazo[4,5-c] quinoline, ENT 1 (enantiomer 1); 1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-8-fluoro-2-(1H-1,2,4-triazol-1-ylmethyl)-1H-imidazo[4,5-с] quinoline, ENT 2 (enantiomer 2); 1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-8-fluoro-2-[(4-methyl-1H-1,2,3-triazol-1-yl)methyl]-1H-imidazo [4,5-c]quinoline, ENT 1 (enantiomer 1); 1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-8-fluoro-2-[(4-methyl-1H-1,2,3-triazol-1-yl)methyl]-1H-imidazo [4,5-c]quinoline, ENT 2 (enantiomer 2); 1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-8-fluoro-2-[(5-methylpyrazin-2-yl)methyl]-1H-imidazo[4,5-c]quinoline, ENT 1 (enantiomer 1); 1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-8-fluoro-2-[(5-methylpyrazin-2-yl)methyl]-1H-imidazo[4,5-c]quinoline, ENT 2 (enantiomer 2); 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-{[4-(methoxymethyl)-1H-1,2,3-triazol-1-yl]methyl}-1H -imidazo[4,5-c]quinoline, ENT 1 (enantiomer 1); 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-{[4-(methoxymethyl)-1H-1,2,3-triazol-1-yl]methyl}-1H -imidazo[4,5-c]quinoline, ENT 2 (enantiomer 2); 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-(1H-1,2,4-triazol-1-ylmethyl)-1H-imidazo[4,5-c] quinoline, ENT 1 (enantiomer 1); 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-(1H-1,2,4-triazol-1-ylmethyl)-1H-imidazo[4,5-c] quinoline, ENT 2 (enantiomer 2); 8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-[(4-methoxy-1H-pyrazol-1-yl)methyl]-1H-imidazo[4,5-с ]quinoline, ENT 1 (enantiomer 1); 8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-[(4-methoxy-1H-pyrazol-1-yl)methyl]-1H-imidazo[4,5-с ]quinoline, ENT 2 (enantiomer 2); 8-fluoro-2-[(2-methylimidazo[2,1-b][1,3,4]thiadiazol-6-yl)methyl]-1-[(2R,4R)-2-methyltetrahydro-2H-pyran -4-yl]-1H-imidazo[4,5-c]quinoline; 2-[(5-methylpyrazin-2-yl)methyl]-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-8-(trifluoromethyl)-1H-imidazo[4,5 -c]quinoline; 2-cyclopentyl-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[4,5-c]quinoline-8-carbonitrile; [cis-4-(8-chloro-2-methyl-1H-imidazo[4,5-c]quinolin-1-yl)tetrahydro-2H-pyran-2-yl]acetonitrile, ENT 1 (enantiomer 1); 1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(5-methyl-1,2,4-oxadiazol-3-yl)methyl]-1H-imidazo[4,5-c ]quinoline-8-carbonitrile, ENT 1 (enantiomer 1); 2-[(5-methylpyrazin-2-yl)methyl]-1-[(3R)-1-methylpyrrolidin-3-yl]-1H-imidazo[4,5-c]quinolin-8-carbonitrile; 1-[(3R)-1-methylpyrrolidin-3-yl]-2-[(5-methyl-2H-tetrazol-2-yl)methyl]-1H-imidazo[4,5-c]quinolin-8-carbonitrile ; 2-[(3-methyl-1,2-oxazol-5-yl)methyl]-1-[(3R)-1-methylpyrrolidin-3-yl]-1H-imidazo[4,5-c]quinoline-8 -carbonitrile; 2-[(4-Methoxy-1H-pyrazol-1-yl)methyl]-1-[(3R)-1-methylpyrrolidin-3-yl]-1H-imidazo[4,5-c]quinolin-8-carbonitrile ; 1-[(3R)-1-methylpyrrolidin-3-yl]-2-[(5-methyl-1,3,4-thiadiazol-2-yl)methyl]-1H-imidazo[4,5-c]quinoline -8-carbonitrile; 2-[(5-methyl-1,3-oxazol-2-yl)methyl]-1-[(3R)-1-methylpyrrolidin-3-yl]-1H-imidazo[4,5-c]quinoline-8 -carbonitrile; 1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-2-{[5-(trifluoromethyl)pyrazin-2-yl]methyl}-1H-imidazo[4,5-с] quinoline-8-carbonitrile; 8-chloro-2-[(6-methylpyrimidin-4-yl)methyl]-1-[(3R)-1-methylpyrrolidin-3-yl]-1H-imidazo[4,5-c]quinoline; 2-[(4-methoxy-1H-pyrazol-1-yl)methyl]-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-8-(trifluoromethyl)-1H-imidazo [4,5-c]quinoline; 8-chloro-2-[(5-methyl-1,2,4-oxadiazol-3-yl)methyl]-1-[(3R)-1-methylpyrrolidin-3-yl]-1H-imidazo[4,5 -c]quinoline; 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-(1H-1,2,4-triazol-1-ylmethyl)-1H-imidazo[4,5-c] quinoline; 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-{[4-(methoxymethyl)-1H-1,2,3-triazol-1-yl]methyl}-1H -imidazo[4,5-c]quinoline; 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(5-methyl-1,2,4-oxadiazol-3-yl)methyl]-1H-imidazo[4 ,5-c]quinoline; 8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-[(4-methyl-1H-1,2,3-triazol-1-yl)methyl]-1H-imidazo [4,5-c]quinoline, ENT 1 (enantiomer 1); 8-chloro-2-[(4-cyclopropyl-1H-1,2,3-triazol-1-yl)methyl]-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-1H-imidazo [4,5-c]quinoline, ENT 2 (enantiomer 2); 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(4-methyl-1H-1,2,3-triazol-1-yl)methyl]-1H-imidazo [4,5-c]quinoline, ENT 2 (enantiomer 2); 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(5-methyl-2H-tetrazol-2-yl)methyl]-1H-imidazo[4,5-с ]quinoline, ENT 1 (enantiomer 1); 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(5-methylpyrazin-2-yl)methyl]-1H-imidazo[4,5-c]quinoline, ENT 1 (enantiomer 1); 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(5-methyl-1,2-oxazol-3-yl)methyl]-1H-imidazo[4,5 -c]quinoline, ENT 1 (enantiomer 1); 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(5-methyl-1,2-oxazol-3-yl)methyl]-1H-imidazo[4,5 -c]quinoline, ENT 2 (enantiomer 2); 1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-8-fluoro-2-{[4-(methoxymethyl)-1H-1,2,3-triazol-1-yl]methyl}-1H -imidazo[4,5-c]quinoline, ENT 2 (enantiomer 2); 8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-(1H-1,2,4-triazol-1-ylmethyl)-1H-imidazo[4,5-c] quinoline, ENT 2 (enantiomer 2); 8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-{[4-(methoxymethyl)-1H-1,2,3-triazol-1-yl]methyl}-1H -imidazo[4,5-c]quinoline, ENT 1 (enantiomer 1); 2-[(5-methyl-1,2,4-oxadiazol-3-yl)methyl]-1-[(3R)-1-(2,2,2-trifluoroethyl)pyrrolidin-3-yl]-1H- imidazo[4,5-c]quinoline-8-carbonitrile; 2-[(4-methoxy-1H-pyrazol-1-yl)methyl]-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[4,5-с ]quinoline-8-carbonitrile; 8-chloro-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-2-(1,3-thiazol-2-ylmethyl)-1H-imidazo[4,5-с] quinoline; 8-chloro-1-[cis-2-(difluoromethyl)tetrahydro-2H-pyran-4-yl]-2-[(5-methyl-1,2-oxazol-3-yl)methyl]-1H-imidazo[ 4,5-c]quinoline, ENT 1 (enantiomer 1); 8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-[(5-methyl-1,2-oxazol-3-yl)methyl]-1H-imidazo[4,5 -c]quinoline, ENT 1 (enantiomer 1); 1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-2-(1,3-thiazol-2-ylmethyl)-1H-imidazo[4,5-c]quinolin-8- carbonitrile; 8-chloro-1-[(3R)-1-methylpyrrolidin-3-yl]-2-[(4-methyl-1H-1,2,3-triazol-1-yl)methyl]-1H-imidazo[4 ,5-c]quinoline; 1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-2-(1,2,3-thiadiazol-4-ylmethyl)-8-(trifluoromethyl)-1H-imidazo[4, 5-c]quinoline; 8-fluoro-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-2-(1,3-thiazol-2-ylmethyl)-1H-imidazo[4,5-с] quinoline; 2-(1,3-benzoxazol-2-ylmethyl)-1-[cis-3-fluorocyclopentyl]-1H-imidazo[4,5-c]quinolin-8-carbonitrile; 1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-2-(1H-1,2,4-triazol-1-ylmethyl)-8-(trifluoromethyl)-1H-imidazo[ 4,5-c]quinoline; 8-chloro-2-[(5-methylpyrazin-2-yl)methyl]-1-[(3R)-1-methylpyrrolidin-3-yl]-1H-imidazo[4,5-c]quinoline; 1-[cis-3-fluorocyclopentyl]-2-[(5-methyl-1,2-oxazol-3-yl)methyl]-1H-imidazo[4,5-c]quinoline-8-carbonitrile; 1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-2-(1,3-thiazol-4-ylmethyl)-8-(trifluoromethyl)-1H-imidazo[4,5- c]quinoline; 2-[(5-methyl-1,3,4-oxadiazol-2-yl)methyl]-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-8-(trifluoromethyl) -1H-imidazo[4,5-c]quinoline; 8-chloro-1-(2,2-dimethyltetrahydro-2H-pyran-4-yl)-2-(1H-1,2,4-triazol-1-ylmethyl)-1H-imidazo[4,5-c] quinoline; 8-chloro-1-(2,2-difluoropropyl)-2-[(4-methoxy-1H-pyrazol-1-yl)methyl]-1H-imidazo[4,5-c]quinoline; 8-fluoro-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-2-{[5-(trifluoromethyl)pyrazin-2-yl]methyl}-1H-imidazo[4, 5-c]quinoline; 1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-2-[(5-methyl-1,3,4-thiadiazol-2-yl)methyl]-8-(trifluoromethyl) -1H-imidazo[4,5-c]quinoline; 8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-[(5-methyl-1,2,4-oxadiazol-3-yl)methyl]-1H-imidazo[4 ,5-c]quinoline, ENT 1 (enantiomer 1); 2-[(6-methylpyrimidin-4-yl)methyl]-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-8-(trifluoromethyl)-1H-imidazo[4,5 -c]quinoline; 8-chloro-1-[cis-3-fluorocyclopentyl]-2-[(5-methylpyrazin-2-yl)methyl]-1H-imidazo[4,5-c]quinoline; 3-{8-chloro-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[4,5-c]quinolin-2-yl}-2-methylpropanenitrile, DIAST 2 (diastereomer 2); 8-fluoro-1-[cis-3-fluorocyclopentyl]-2-(1,2,3-thiadiazol-4-ylmethyl)-1H-imidazo[4,5-c]quinoline, ENT 2 (enantiomer 2); 3-{8-chloro-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[4,5-c]quinolin-2-yl}propanenitrile; 1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-2-[(5-methyl-2H-tetrazol-2-yl)methyl]-8-(trifluoromethyl)-1H-imidazo [4,5-c]quinoline; 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(4-methyl-1H-1,2,3-triazol-1-yl)methyl]-1H-imidazo [4,5-c]quinoline; 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-(1H-tetrazol-1-ylmethyl)-1H-imidazo[4,5-c]quinoline; 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(1-methyl-1H-1,2,4-triazol-3-yl)methyl]-1H-imidazo [4,5-c]quinoline; 1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-8-fluoro-2-[(2-methylimidazo[2,1-b][1,3,4]thiadiazol-6-yl)methyl ]-1H-imidazo[4,5-c]quinoline, ENT 1 (enantiomer 1); 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-(1H-tetrazol-1-ylmethyl)-1H-imidazo[4,5-c]quinoline, ENT 2 (enantiomer 2); 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(4-methyl-2H-1,2,3-triazol-2-yl)methyl]-1H-imidazo [4,5-c]quinoline, ENT 1 (enantiomer 1); 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(5-methyl-1,2,4-oxadiazol-3-yl)methyl]-1H-imidazo[4 ,5-c]quinoline, ENT 1 (enantiomer 1); 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(5-methyl-1,3,4-thiadiazol-2-yl)methyl]-1H-imidazo[4 ,5-c]quinoline, ENT 2 (enantiomer 2); 8-(difluoromethyl)-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-2-[(4-methyl-1H-1,2,3-triazol-1-yl) methyl]-1H-imidazo[4,5-c]quinoline; 8-(difluoromethyl)-2-[(5-methyl-1,2-oxazol-3-yl)methyl]-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H -imidazo[4,5-c]quinoline; 8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-[(5-methyl-1,3,4-thiadiazol-2-yl)methyl]-1H-imidazo[4 ,5-c]quinoline, ENT 1 (enantiomer 1); 8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-(1H-1,2,4-triazol-1-ylmethyl)-1H-imidazo[4,5-c] quinoline, ENT 1 (enantiomer 1); 8-chloro-2-[(4-cyclopropyl-1H-1,2,3-triazol-1-yl)methyl]-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-1H-imidazo [4,5-s]quinoline and [5-({8-chloro-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[4,5-c]quinolin-2-yl}methyl)pyrazine -2-yl]methanol, or a pharmaceutically acceptable salt thereof, wherein the compound or pharmaceutically acceptable salt optionally has one or more hydrogen atoms that have been replaced by a deuterium atom. 3. The compound according to claim 2, which is selected from the group consisting of: 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-{[4-(methoxymethyl)-1H-1,2,3-triazol-1-yl]methyl}-1H -imidazo[4,5-c]quinoline, ENT 2 (enantiomer 2); 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-(1H-1,2,4-triazol-1-ylmethyl)-1H-imidazo[4,5-c] quinoline, ENT 1 (enantiomer 1); 2-[(6-methylpyrimidin-4-yl)methyl]-1-[(3R)-1-methylpyrrolidin-3-yl]-1H-imidazo[4,5-c]quinolin-8-carbonitrile; 8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-[(5-methylpyrazin-2-yl)methyl]-1H-imidazo[4,5-c]quinoline, ENT 2 (enantiomer 2); 1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(4-methyl-2H-1,2,3-triazol-2-yl)methyl]-1H-imidazo[4,5 -c]quinoline-8-carbonitrile, ENT 2 (enantiomer 2); 1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-2-[(1-methyl-1H-1,2,3-triazol-4-yl)methyl]-8-( trifluoromethyl)-1H-imidazo[4,5-c]quinoline; [(2S,4R)-4-(8-chloro-2-ethyl-1H-imidazo[4,5-c]quinolin-1-yl)tetrahydro-2H-pyran-2-yl]acetonitrile; 8-(difluoromethyl)-2-[(5-methylpyrazin-2-yl)methyl]-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[4,5 -c]quinoline; 8-chloro-1-[(4S)-3,3-difluorotetrahydro-2H-pyran-4-yl]-2-[(5-methyl-1,2-oxazol-3-yl)methyl]-1H-imidazo [4,5-c]quinoline; 1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(5-methyl-1,2,4-oxadiazol-3-yl)methyl]-1H-imidazo[4,5-c ]quinoline-8-carbonitrile, ENT 1 (enantiomer 1); 8-fluoro-2-[(2-methylimidazo[2,1-b][1,3,4]thiadiazol-6-yl)methyl]-1-[(2R,4R)-2-methyltetrahydro-2H-pyran -4-yl]-1H-imidazo[4,5-c]quinoline; 2-[(5-methylpyrazin-2-yl)methyl]-1-[(3R)-1-methylpyrrolidin-3-yl]-1H-imidazo[4,5-c]quinolin-8-carbonitrile; 1-[(3R)-1-methylpyrrolidin-3-yl]-2-[(5-methyl-1,3,4-thiadiazol-2-yl)methyl]-1H-imidazo[4,5-c]quinoline -8-carbonitrile; 1-[(3R)-1-methylpyrrolidin-3-yl]-2-[(5-methyl-2H-tetrazol-2-yl)methyl]-1H-imidazo[4,5-c]quinolin-8-carbonitrile ; 2-[(5-methylpyrazin-2-yl)methyl]-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-8-(trifluoromethyl)-1H-imidazo[4,5 -c]quinoline; [cis-4-(8-chloro-2-cyclobutyl-1H-imidazo[4,5-c]quinolin-1-yl)tetrahydro-2H-pyran-2-yl]acetonitrile, ENT 2 (enantiomer 2); 1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-8-fluoro-2-[(5-methylpyrazin-2-yl)methyl]-1H-imidazo[4,5-c]quinoline, ENT 1 (enantiomer 1); 1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-8-fluoro-2-[(4-methyl-1H-1,2,3-triazol-1-yl)methyl]-1H-imidazo [4,5-c]quinoline, ENT 2 (enantiomer 2) and 1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-8-fluoro-2-(1H-1,2,4-triazol-1-ylmethyl)-1H-imidazo[4,5-c] quinoline, ENT 2 (enantiomer 2); or a pharmaceutically acceptable salt thereof, wherein the compound or pharmaceutically acceptable salt optionally has one or more hydrogen atoms that have been replaced by a deuterium atom. 4. The compound according to claim 3, which is selected from the group consisting of: 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-{[4-(methoxymethyl)-1H-1,2,3-triazol-1-yl]methyl}-1H -imidazo[4,5-c]quinoline, ENT 2 (enantiomer 2); 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-(1H-1,2,4-triazol-1-ylmethyl)-1H-imidazo[4,5-c] quinoline, ENT 1 (enantiomer 1); 2-[(6-methylpyrimidin-4-yl)methyl]-1-[(3R)-1-methylpyrrolidin-3-yl]-1H-imidazo[4,5-c]quinolin-8-carbonitrile; 8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-[(5-methylpyrazin-2-yl)methyl]-1H-imidazo[4,5-c]quinoline, ENT 2 (enantiomer 2); 1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(4-methyl-2H-1,2,3-triazol-2-yl)methyl]-1H-imidazo[4,5 -c]quinoline-8-carbonitrile, ENT 2 (enantiomer 2); 1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-2-[(1-methyl-1H-1,2,3-triazol-4-yl)methyl]-8-( trifluoromethyl)-1H-imidazo[4,5-c]quinoline; [(2S,4R)-4-(8-chloro-2-ethyl-1H-imidazo[4,5-c]quinolin-1-yl)tetrahydro-2H-pyran-2-yl]acetonitrile; 8-(difluoromethyl)-2-[(5-methylpyrazin-2-yl)methyl]-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[4,5 -c]quinoline; 8-chloro-1-[(4S)-3,3-difluorotetrahydro-2H-pyran-4-yl]-2-[(5-methyl-1,2-oxazol-3-yl)methyl]-1H-imidazo [4,5-s]quinoline and 1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(5-methyl-1,2,4-oxadiazol-3-yl)methyl]-1H-imidazo[4,5-c ]quinoline-8-carbonitrile, ENT 1 (enantiomer 1); or a pharmaceutically acceptable salt thereof, wherein the compound or pharmaceutically acceptable salt optionally has one or more hydrogen atoms that have been replaced by a deuterium atom. 5. The compound according to claim 4, which is selected from the group consisting of: 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-{[4-(methoxymethyl)-1H-1,2,3-triazol-1-yl]methyl}-1H -imidazo[4,5-c]quinoline, ENT 2 (enantiomer 2); 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-(1H-1,2,4-triazol-1-ylmethyl)-1H-imidazo[4,5-c] quinoline, ENT 1 (enantiomer 1); 2-[(6-methylpyrimidin-4-yl)methyl]-1-[(3R)-1-methylpyrrolidin-3-yl]-1H-imidazo[4,5-c]quinolin-8-carbonitrile; 8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-[(5-methylpyrazin-2-yl)methyl]-1H-imidazo[4,5-c]quinoline, ENT 2 (enantiomer 2) and 1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(4-methyl-2H-1,2,3-triazol-2-yl)methyl]-1H-imidazo[4,5 -c]quinoline-8-carbonitrile, ENT 2 (enantiomer 2); or a pharmaceutically acceptable salt thereof, wherein the compound or pharmaceutically acceptable salt optionally has one or more hydrogen atoms that have been replaced by a deuterium atom. 6. The compound according to claim 5, which is 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-{[4-(methoxymethyl)-1H-1,2,3- triazol-1-yl]methyl}-1H-imidazo[4,5-c]quinoline, ENT 2 (enantiomer 2); or a pharmaceutically acceptable salt thereof, wherein the compound or pharmaceutically acceptable salt optionally has one or more hydrogen atoms that have been replaced by a deuterium atom. 7. The compound according to claim 5, which is 8-chloro-1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-(1H-1,2,4-triazol-1-ylmethyl)- 1H-imidazo[4,5-c]quinoline, ENT 1 (enantiomer 1); or a pharmaceutically acceptable salt thereof, wherein the compound or pharmaceutically acceptable salt optionally has one or more hydrogen atoms that have been replaced by a deuterium atom. 8. The compound according to claim 5, which is 2-[(6-methylpyrimidin-4-yl)methyl]-1-[(3R)-1-methylpyrrolidin-3-yl]-1H-imidazo[4,5-c ]quinoline-8-carbonitrile; or a pharmaceutically acceptable salt thereof, wherein the compound or pharmaceutically acceptable salt optionally has one or more hydrogen atoms that have been replaced by a deuterium atom. 9. The compound according to claim 5, which is 8-chloro-1-(3,3-difluorotetrahydro-2H-pyran-4-yl)-2-[(5-methylpyrazin-2-yl)methyl]-1H-imidazo [4,5-c]quinoline, ENT 2 (enantiomer 2); or a pharmaceutically acceptable salt thereof, wherein the compound or pharmaceutically acceptable salt optionally has one or more hydrogen atoms that have been replaced by a deuterium atom. 10. The compound according to claim 5, which is 1-(4,4-difluoro-1-methylpyrrolidin-3-yl)-2-[(4-methyl-2H-1,2,3-triazol-2-yl) methyl]-1H-imidazo[4,5-c]quinoline-8-carbonitrile, ENT 2 (enantiomer 2); or a pharmaceutically acceptable salt thereof, wherein the compound or pharmaceutically acceptable salt optionally has one or more hydrogen atoms that have been replaced by a deuterium atom. 11. The compound according to claim 1, where R ¹ is ethyl,

R ² is

R ³ is chlorine, cyano, difluoromethyl or trifluoromethyl, or a pharmaceutically acceptable salt thereof, wherein the compound or pharmaceutically acceptable salt has one or more hydrogen atoms which is replaced by a deuterium atom, provided that one or more hydrogen atoms are not replaced by only a deuterium atom in one or more of the 2 positions, 5, 7 or 8 quinoline ring. 12. Connection according to claim 1, where R ¹ is

R ² is

R ³ is chlorine or cyano, or a pharmaceutically acceptable salt thereof, wherein the compound or pharmaceutically acceptable salt has one or more hydrogen atoms which is replaced by a deuterium atom, provided that one or more hydrogen atoms are not replaced by only a deuterium atom in one or more of the 2 positions, 5, 7 or 8 quinoline ring. 13. The compound according to claim 1, where R ¹ is

and R ² is

and R ³ is chlorine, or a pharmaceutically acceptable salt thereof. 14. Compound [5-({8-chloro-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H-imidazo[4,5-c]quinolin-2-yl} methyl)pyrazin-2-yl]methanol or a pharmaceutically acceptable salt thereof, wherein the compound or pharmaceutically acceptable salt optionally has one or more hydrogen atoms that have been replaced by a deuterium atom. 15. Compound 8-chloro-2-{[5-( ² H ₃ )methylpyrazin-2-yl]methyl}-1-[(2R,4R)-2-methyltetrahydro-2H-pyran-4-yl]-1H -imidazo[4,5-c]quinoline or a pharmaceutically acceptable salt thereof. 16. Pharmaceutical composition having inhibitory activity against leucine-rich repeat kinase 2 (LRRK2), containing a therapeutically effective amount of a compound according to any one of paragraphs. 1-15 or a pharmaceutically acceptable salt thereof, together with a pharmaceutically acceptable carrier. 17. A pharmaceutical composition according to claim 16 for use in the treatment of a disease or disorder selected from the group consisting of Crohn's disease, Parkinson's disease, dementia with Lewy bodies, leprosy, Alzheimer's disease and tauopathy. 18. A method of treating a disease or disorder selected from the group consisting of Crohn's disease, Parkinson's disease, Lewy body dementia, leprosy, Alzheimer's disease, and tauopathy in a patient, said method comprising administering to a patient in need of such treatment a therapeutically effective amount a compound or a pharmaceutically acceptable salt thereof according to any one of paragraphs. 1-15 or a pharmaceutical composition according to claim 16. 19. The compound or its pharmaceutically acceptable salt according to any one of paragraphs. 1-15 for use in the treatment of a disease or disorder selected from the group consisting of Crohn's disease, Parkinson's disease, Lewy body dementia, leprosy, Alzheimer's disease, and tauopathy.