Phenyl-3,4-dihydroisoquinoline-2(1H)-yl-ethyl-1-one derivatives that act as orthoallosteric regulators of dopamine D1 receptors

Abstract

This invention provides certain compounds of formula I (phenyl)-3,4-dihydroisoquinoline-2(1H)-yl)ethyl-1-one and pharmaceutical compositions thereof as D1 allosteric modulators (PAM). This invention also provides a method for treating certain symptoms of dopaminergic CNS disorders using compounds of formula I, including Parkinson's disease, schizophrenia, ADHD, or Alzheimer's disease.

Description

[0001] This invention provides certain phenyl-3,4-dihydroisoquinoline-2(1H)-yl-ethyl-1-one related compounds, pharmaceutical compositions thereof, and methods for using them to treat dopaminergic CNS disorders, including Parkinson's disease, Alzheimer's disease, schizophrenia, and attention deficit hyperactivity disorder (ADHD).

[0002] Many currently used drugs act directly or indirectly through dopamine receptors. These include dopamine agonists and the dopamine precursor L-DOPA for Parkinson's disease, dopamine releasers for attention deficit disorder and narcolepsy, and dopamine reuptake inhibitors for depression. D1 receptors play a crucial role in motor activity and reward, and have a specific role in maintaining higher cognitive functions such as working memory, attention, and executive function (Arnsten AF, Cereb. Cortex (2013) 123, 2269-2281). To date, attempts to develop D1 agonists for clinical application have been unsuccessful, prompting the search for alternative methods to increase D1 receptor activity.

[0003] One approach is to identify allosteric potentiators of the dopamine D1 receptor, also known as positive allosteric modulators or PAMs (Svensson K et al., J. Pharmacol. Exp. Ther. (2017) 360: 117–128). Allosteric modulators are agents that enhance (positive allosteric modulators or PAMs) or inhibit (negative allosteric modulators or NAMs) the action of natural ligands by binding to sites different from the allosteric binding site on the receptor (allosteric binding sites). By increasing dopamine's affinity for the D1 receptor, D1 potentiators amplify the response to endogenous dopamine, increasing D1 tone at and at dopamine release. This mode of activity is the opposite of that of D1 agonists, which activate all D1 receptors they can access in the presence of D1 agonists. In animal models of cognitive and motor activity, D1 agonists exhibit a bell-shaped dose-response relationship, likely due to overstimulation at higher doses. Due to the constant activation of the D1 receptor, some D1 agonists also show rapid development of tolerance. Conversely, because D1 enhancers depend on endogenous tone and are subject to normal feedback control, they may have a much lower tendency to overstimulate. Given that dopamine and D1 receptor signaling are involved in these central nervous system functions, D1 enhancers that enhance D1 receptor activity could provide alternatives and / or ameliorative agents for the treatment of certain dopamine-related disorders.

[0004] Parkinson's disease is a chronic, progressive neurodegenerative disorder characterized by the loss of dopaminergic neurons in the brain. Parkinson's disease manifests as resting tremor and other motor symptoms (such as bradykinesia and postural instability) and nonmotor symptoms (such as cognitive impairment, sleep disturbances, and depression). Current treatments for Parkinson's disease include the administration of non-selective dopamine precursors such as levodopa and dopamine receptor agonists. Direct-acting dopamine receptor agonist therapy may also be associated with impulse control disorders, psychosis, and cognitive decline due to their relatively high affinity for D2 receptors. Schizophrenia is a debilitating illness with a complex pathological mechanism. One component of schizophrenia is cognitive impairment, which may be related to a lack of D1 receptor activation or downregulation. It has been hypothesized that more selective D1 activation relative to D2 regulation could be effective in treating cognitive impairment associated with schizophrenia. Alzheimer's disease is a chronic, progressive neurodegenerative disorder characterized by the loss of neurons and synapses in the cerebral cortex and certain subcortical regions. Disease progression includes cognitive impairment, which is hypothesized to be at least partly due to reduced D1 receptor activation; therefore, D1 activation may provide therapeutic benefit in treating cognitive impairment associated with Alzheimer's disease. ADHD is a neurodevelopmental disorder characterized by difficulty concentrating, hyperactivity, or difficulty controlling behavior according to a person's age. It is hypothesized that D1 activation may provide therapeutic benefit in treating ADHD. Therefore, a significant unmet need remains for safe and effective treatment of cognitive or other neurological deficits associated with schizophrenia, Parkinson's disease, Alzheimer's disease, and / or ADHD (e.g., alternative and / or modified dopamine D1 receptor orthoallosteric modulators (D1 PAM)).

[0005] WO 2014 / 193781 describes certain 3,4-dihydroisoquinoline-2(1H)-yl compounds as D1 PAM for the treatment of cognitive impairment associated with Parkinson's disease, Alzheimer's disease, schizophrenia, depression, or ADHD.

[0006] This invention provides novel compounds that are selective PAMs for the dopamine 1 receptor (D1) and exhibit a favorable combination of pharmacological properties, such as enhanced human D1 receptor signaling in response to dopamine, high oral in vivo availability, and in vivo efficacy in animals accustomed to their environment and activating motor function. Therefore, the compounds of this invention are considered for the treatment of Parkinson's disease, Alzheimer's disease, schizophrenia, and / or ADHD. The compounds of this invention can provide alternative treatments for such conditions.

[0007] This invention provides a compound having Formula I:

[0008]

[0009] in:

[0010] R 1 yes

[0011] R 2 It is -F or –Cl;

[0012] R 3 It is -F or –Cl; and

[0013] R 4 It is -H or -F;

[0014] The condition is when R 1 for When R4 is -F, then R4 is -F.

[0015] Compounds having Formula I are particularly useful in the treatment methods of the present invention, but certain configurations are preferred. Such preferred configurations are described in the following paragraphs. Although the invention as embodied in Formula I contemplates all individual enantiomers and diastereomers of the said compounds, as well as mixtures of enantiomers and / or diastereomers, including racemic mixtures, compounds having the absolute configurations described below are preferred. It should be understood that these preferred embodiments apply to the treatment methods and novel compounds of the present invention.

[0016] A particular embodiment of Formula I is a compound having Formula Ia:

[0017]

[0018] in:

[0019] R 1 yes

[0020] R 2 It is -F or –Cl;

[0021] R 3 It is -F or –Cl; and

[0022] R 4 It is -H or -F;

[0023] The condition is when R 1 for When R4 is -F, then R4 is -F.

[0024] A particular embodiment of Formula I is a compound having Formula Ib:

[0025]

[0026] Its free base form can also be named 2-(2-chloro-6-fluoro-phenyl)-1-[(1S,3R)-6-fluoro-3-(hydroxymethyl)-5-(3-hydroxy-3-methyl-butyl)-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl] ethyl ketone.

[0027] Furthermore, the present invention provides a pharmaceutical composition comprising a compound having formula I, Ia and / or Ib, and a pharmaceutically acceptable carrier, diluent or excipient.

[0028] The following specific examples are compounds having formula I or Ia.

[0029] This invention provides a compound, which is

[0030]

[0031] This invention provides a compound, which is

[0032]

[0033] This invention provides a compound, which is

[0034]

[0035] This invention provides a compound, which is

[0036]

[0037] This invention provides a compound, which is

[0038]

[0039] This invention provides a compound, which is

[0040]

[0041] This invention provides a compound, which is

[0042]

[0043] This invention provides a compound, which is

[0044]

[0045] This invention provides a compound, which is

[0046]

[0047] This invention provides a compound, which is

[0048]

[0049] Furthermore, the present invention provides compounds of one of the specific embodiments just listed above, and pharmaceutically acceptable carriers, diluents, or excipients. For example, the compound embodiments described herein contemplate co-crystallization.

[0050] The compounds of the present invention are selective PAMs of dopamine 1 (D1) receptors with minimal activity against dopamine 2 (D2) receptors. The compounds of the present invention can further provide their therapeutic benefits while avoiding the risk of drug-drug interactions. Therefore, the compounds of the present invention are considered for use in treating conditions in which reducing D1 activity is effective and D2 activation is not desired, such as Parkinson's disease and schizophrenia, including relief of certain associated symptoms, such as motor symptoms and cognitive deficits associated with Parkinson's disease, and cognitive deficits and negative symptoms associated with schizophrenia, such as mild cognitive impairment or dementia. The compounds of the present invention are also considered for use as a monotherapy or in combination with other therapies to improve motor symptoms of Parkinson's disease. The compounds of the present invention are also considered for use in treating certain symptoms of Alzheimer's disease, such as cognitive deficits, such as mild cognitive impairment. Furthermore, the compounds of the present invention are considered for use in treating certain symptoms of ADHD. The compounds of the present invention are also believed to be useful in treating dopaminergic CNS disorders, including Parkinson's disease, Alzheimer's disease, Lewy body dementia (LBD), vascular dementia, schizophrenia, ADHD, depression, autism, chronic musculoskeletal pain, fibromyalgia, cognitive impairment, sleep disorders, excessive daytime sleepiness, narcolepsy, shift work disorder, traumatic brain injury, chronic traumatic encephalopathy, obesity and appetite regulation, mood disorders, lethargy, apathy, and addiction disorders.

[0051] Furthermore, the present invention provides a compound having formula I, which is used for treatment. Furthermore, the present invention provides a compound having formula Ia, which is used for treatment. Furthermore, the present invention provides a compound having formula Ib, which is used for treatment.

[0052] On the other hand, the present invention provides a pharmaceutical composition comprising a compound having formula I, Ia, or Ib, and one or more pharmaceutically acceptable excipients, diluents, or carriers. Furthermore, this aspect of the invention provides a pharmaceutical composition for treating Parkinson's disease, such as cognitive impairment associated with Parkinson's disease, comprising a compound having formula I, Ia, or Ib, and one or more pharmaceutically acceptable excipients, carriers, or diluents. In another embodiment of this aspect of the invention, a pharmaceutical composition for alleviating motor impairment associated with Parkinson's disease is provided, comprising a compound having formula I, Ia, or Ib, and one or more pharmaceutically acceptable excipients, carriers, or diluents.

[0053] In another embodiment of this aspect of the invention, a pharmaceutical composition for treating Alzheimer's disease, such as alleviating cognitive deficits associated with Alzheimer's disease, is provided, comprising a compound having formula I, Ia, or Ib, and one or more pharmaceutically acceptable excipients, carriers, or diluents.

[0054] Another embodiment of this aspect of the invention provides a pharmaceutical composition for treating schizophrenia, such as reducing cognitive deficits associated with schizophrenia, comprising a compound having formula I, Ia, or Ib, and one or more pharmaceutically acceptable excipients, carriers, or diluents.

[0055] Another embodiment of the present invention provides a pharmaceutical composition for treating ADHD, comprising a compound having formula I, Ia or Ib, and one or more pharmaceutically acceptable excipients, carriers or diluents.

[0056] Furthermore, the present invention provides a method for treating Parkinson's disease, such as cognitive impairment associated with Parkinson's disease, or for reducing motor impairment associated with Parkinson's disease, comprising administering to a patient in need an effective amount of a compound having formula I, Ia, or Ib.

[0057] Furthermore, the present invention provides a method for treating Alzheimer's disease, such as cognitive impairment associated with Alzheimer's disease, comprising administering to a patient in need an effective amount of a compound having formula I, Ia, or Ib.

[0058] Furthermore, the present invention provides a method for treating schizophrenia, such as cognitive impairment associated with schizophrenia, comprising administering to a patient in need an effective amount of a compound having formula I, Ia, or Ib.

[0059] Furthermore, the present invention provides a method for treating ADHD, comprising administering to a patient in need an effective amount of a compound having formula I, Ia, or Ib.

[0060] In one embodiment of this aspect, the present invention provides a compound having formula I, Ia, or Ib for treating Parkinson's disease. In a specific embodiment, the present invention provides a compound having formula I, Ia, or Ib for treating cognitive impairment associated with Parkinson's disease. In another specific embodiment, the present invention provides a compound having formula I, Ia, or Ib for alleviating motor impairment associated with Parkinson's disease.

[0061] Furthermore, the present invention provides a compound having formula I, Ia, or Ib for treating schizophrenia, such as for treating cognitive impairment associated with schizophrenia.

[0062] Furthermore, the present invention provides a compound having formula I, Ia, or Ib for the treatment of ADHD.

[0063] Furthermore, the present invention provides a compound having formula I, Ia or Ib for treating Alzheimer's disease, such as for treating cognitive impairment associated with Alzheimer's disease.

[0064] In another aspect, the present invention provides the use of compounds having formula I, Ia or Ib in the preparation of medicaments for treating Parkinson's disease, such as treating cognitive impairment associated with Parkinson's disease or alleviating motor impairment associated with Parkinson's disease.

[0065] Furthermore, the present invention provides the use of compounds having formula I, Ia, or Ib in the preparation of medicaments for treating schizophrenia, such as for treating cognitive impairment associated with schizophrenia.

[0066] Furthermore, the present invention provides the use of compounds having formula I, Ia, or Ib in the preparation of medicaments for treating Alzheimer's disease, such as for treating cognitive impairment associated with Alzheimer's disease.

[0067] Furthermore, the present invention provides the use of compounds having formula I, Ia, or Ib in the preparation of medicaments for treating ADHD.

[0068] Although the compounds used in the methods of this invention can be administered directly without any formulation, the compounds are typically administered in the form of pharmaceutical compositions comprising a compound of formula I, Ia, or Ib as the active ingredient, and at least one pharmaceutically acceptable carrier, diluent, and / or excipient. These compositions can be administered via a variety of routes, including oral, sublingual, nasal, subcutaneous, intravenous, and intramuscular. Such pharmaceutical compositions and methods of their preparation are well known in the art. See, for example, Remington: The Science and Practice of Pharmacy (University of the Sciences in Philadelphia, ed., 21st ed., Lippincott Williams & Wilkins Co., 2005).

[0069] The compositions are preferably formulated as unit dosage forms, each containing about 0.5 mg to about 800 mg of the active ingredient. The term "unit dosage form" refers to a physically discrete unit suitable as a unit dose for use in human subjects and other mammals, each unit containing a predetermined amount of active substance calculated to produce the desired therapeutic effect, and at least one suitable pharmaceutically acceptable carrier, diluent, and / or excipient. It should be understood that the amount of compound actually administered will be determined by a physician based on relevant circumstances, including the condition to be treated, the chosen route of administration, the actual compound administered, the individual patient's age, weight, and response, and the severity of the patient's symptoms. It is considered that the compounds of the present invention (e.g., in the pharmaceutical compositions of the present invention) will be used for the treatment of Alzheimer's disease, Parkinson's disease, and / or schizophrenia, such as mild cognitive impairment associated with these diseases, through long-term administration.

[0070] The compounds of the present invention can be used in combination with other drugs for the treatment or improvement of dopaminergic CNS disorders. One or more such other drugs may be administered concurrently or sequentially with the compounds of the present invention via the usual route and dosage. For example, other active ingredients that can be combined with the compounds of the present invention for the effective treatment of Parkinson's disease include, but are not limited to: (A) dopamine precursors such as levodopa; melevodopa and etilevodopa; and (b) dopamine agonists, including pramipexole, ropinirole, apomorphine, rotigotine, bromocriptine, cabergoline, and pergolid. (e) Monoamine oxidase inhibitors, including selegiline and rasagiline; (d) COMT inhibitors, including tolcapone and entacapone; (e) Acetylcholinesterase inhibitors, including rivastigmine and donepezil; (f) Antidepressants, including sertraline, citalopram, mirtazapine, and trazodone. In addition, other active ingredients that can be combined with the compounds of the present invention for the effective treatment of Alzheimer's disease include, but are not limited to: acetylcholinesterase inhibitors, including levamisole and donepezil; NMDA antagonists, including memantine; certain antipsychotics, including risperidone, quetiapine, aripiprazole, olanzapine, and pimavanserin; and antidepressants, including sertraline, citalopram, mirtazapine, and trazodone.

[0071] As used herein, the term "patient" refers to a mammal, such as a human, that requires treatment for a condition or disease. Humans are preferred patients. As used herein, patients treated with the compounds of the present invention suffer from dopaminergic CNS disorders and thus share an etiological aspect, since dopamine signaling disorders are known to contribute to these diseases. Dopaminergic CNS disorders as used herein include Parkinson's disease, Alzheimer's disease, Lewy body dementia (LBD), vascular dementia, schizophrenia, ADHD, depression, autism, chronic musculoskeletal pain, fibromyalgia, cognitive impairment, sleep disorders, excessive daytime sleepiness, narcolepsy, shift work disorder, traumatic brain injury, chronic traumatic encephalopathy, obesity and appetite regulation, mood disorders, somnolence, emotional blunting, and addiction disorders. Identification of patients with these dopaminergic CNS disorders can be achieved through established methods known to those skilled in the art.

[0072] This invention provides a method for treating patients with dopaminergic diseases by simultaneously, separately, or sequentially combining a compound having formula I, Ia, or Ib with a dopamine precursor. This invention also provides a method for treating patients with Parkinson's disease by simultaneously, separately, or sequentially combining a compound having formula I, Ia, or Ib with a dopamine precursor.

[0073] This invention provides a method for treating patients with Alzheimer's disease by combining a compound having formula I, Ia, or Ib with a dopamine precursor simultaneously, separately, or sequentially.

[0074] This invention provides a method for treating patients with dopaminergic disorders by simultaneously, separately, or sequentially combining a compound having formula I, Ia, or Ib with a dopamine agonist. This invention also provides a method for treating patients with Parkinson's disease by simultaneously, separately, or sequentially combining a compound having formula I, Ia, or Ib with a dopamine agonist.

[0075] This invention provides a method for treating patients with Alzheimer's disease by combining a compound having formula I, Ia, or Ib with a dopamine agonist simultaneously, separately, or sequentially.

[0076] In embodiments of the invention, the patient is someone diagnosed with a medical risk, symptom, or condition, such as dopaminergic CNS disorder, requiring treatment with the dosing regimen described herein. In cases where conditions treatable by the methods of the invention are known through established and accepted classifications such as AD, PD, and LBD, their classifications can be found in various well-known medical texts. For example, the fifth edition of the Diagnostic and Statistical Manual of Mental Disorders (DSM-5) currently provides diagnostic tools for identifying many of the conditions described herein. Furthermore, the tenth revision of the International Classification of Diseases (ICD-10) provides classifications for many of the conditions described herein. Those skilled in the art will recognize that alternative nomenclature, disease taxonomy, and classification systems exist for the conditions described herein, including those described in DSM-5 and ICD-10, and that terminology and classification systems evolve with advancements in medical science. Cognitive impairment in subjects with Parkinson's disease is commonly referred to as a neurocognitive disorder. The diagnostic criteria in DSM-5 (Diagnostic and Statistical Manual of Mental Disorders, 5th Edition) describe evidence of a significant cognitive decline from previous performance levels (concern recorded by neuropsychological testing or by the notifying person), and that the cognitive deficit may or may not interfere with the independence of daily activities. This is considered severe or mild neurocognitive disorder. As used in this article, “weight loss” refers to weight reduction and / or chronic weight management in which treatment promotes maintaining weight within a desired range.

[0077] As used herein, the terms “treatment / treating” or “mitigating” are intended to refer to all processes that can slow, interrupt, prevent, control, or stop the progression of an existing condition and / or the reduction of its symptoms, but do not necessarily indicate the complete elimination of all symptoms.

[0078] As used herein, the term "effective amount" of a compound having formula I, Ia, or Ib refers to the amount, i.e., the dose, that effectively enhances a dopamine-mediated response in a patient. A preferred "effective amount" can be determined as the amount that promotes a patient's state of arousal or alertness compared to an untreated patient. In determining the effective amount or dose of a compound having formula I, Ia, or Ib, many factors are considered, including but not limited to the compound to be administered and its specific formulation; the patient's body type, age, and general health condition; the extent or severity of the disease involved; the individual patient's response; the manner of administration; and other relevant circumstances.

[0079] The abbreviations used in this article are defined as follows:

[0080] “Bn” refers to benzyl.

[0081] “CAS#” refers to the Chemical Abstracts Registry Number.

[0082] "Crotonyl" refers to but-2-en-1-yl.

[0083] "CNS" refers to the central nervous system.

[0084] "1D-NOESY" refers to the one-dimensional nuclear overhauser effect NMR spectrum.

[0085] "DCM" refers to dichloromethane or dichloromethane.

[0086] "DIPEA" refers to N,N-diisopropylethylamine.

[0087] “dmso-d6” refers to fully deuterated dimethyl sulfoxide.

[0088] “ee” refers to an enantiomer excess.

[0089] "ESMS" refers to electrospray mass spectrometry.

[0090] “EtOAc” refers to ethyl acetate.

[0091] “h” refers to hours or number of hours.

[0092] “HATU” refers to 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate or N-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethylammonium hexafluorophosphate N-oxide.

[0093] "HPLC" refers to High Performance Liquid Chromatography.

[0094] “LAH” refers to lithium aluminum hydride.

[0095] “MeOH” refers to methanol or methyl alcohol.

[0096] “min” refers to minutes or minutes.

[0097] "NMR" refers to nuclear magnetic resonance spectroscopy.

[0098] “PG” refers to the protective base.

[0099] “Ph” refers to phenyl.

[0100] “OAc” refers to acetate.

[0101] “rac-” refers to racemic or racemic compounds.

[0102] “RT” refers to room temperature or ambient temperature.

[0103] “SFC” refers to supercritical fluid chromatography.

[0104] "TBDMS" refers to tert-butyldimethylsilyl alkyl.

[0105] "TBDPS" refers to tert-butyldiphenylsilyl alkyl.

[0106] "THF" refers to tetrahydrofuran.

[0107] “t R "" refers to the retention period.

[0108] "w / w" refers to weight to weight or weight ratio as a unit of mass.

[0109] “wt%” refers to weight percentage.

[0110] “Z” refers to benzyloxycarbonyl group, -C(O)OCH2Ph, which serves as a protecting group.

[0111] General Chemistry

[0112] The compounds of the present invention can be prepared by general methods known and understood in the art or by the methods described herein. Suitable reaction conditions for the steps used in these schemes are well known in the art, and appropriate substitutions of solvents and co-reagents are known to those skilled in the art. Similarly, those skilled in the art will understand that synthetic intermediates can be isolated and / or purified as needed or desired by various well-known techniques, and often, various intermediates can be used directly in subsequent synthetic steps with little or no purification. Furthermore, those skilled in the art will understand that in some cases, the order of introduction of the components is not critical.

[0113] Option 1

[0114]

[0115] Scheme 1 describes the preparation of compound 8. Those skilled in the art will recognize suitable disubstituted phenylalanine 1 (e.g., X = Br, Cl, I; R) dissolved in polar protic solvents. 4=H,F), can be obtained by esterification with a suitable strong acid to obtain esterified salt 2. The salt is then washed with an aqueous alkaline solution to obtain a free base, dissolved in an aprotic solvent, and acylated with a suitable acyl chloride to obtain 3. The 2-substituted N-acylated phenylalanine methyl ester 3 is known in the art to be cyclized by treatment with paraformaldehyde in a suitable strong acid and stirring to obtain tetrahydroisoquinoline 4. Demethylation and decarboxylation can be achieved by treatment with an aqueous acid solution and stirring under reflux to obtain 5 as the corresponding amine salt. Those skilled in the art will recognize that N-protected tetrahydroisoquinoline 6 can be formed by dissolving amine salt 5 in a suitable polar aprotic solvent, adding a base and a suitable acid anhydride or alkyl chloroformate to obtain tert-butyl carbamate 6. Subsequent reduction to methanol derivative 7 can be carried out using a series of reducing agents, such as metal hydrides, borohydride salts, or diborane in a polar aprotic solvent. O-protected tetrahydroisoquinoline 8 can be achieved by first treating N-protected tetrahydroisoquinoline 7 with a suitable strong acid and concentrating under vacuum. Subsequently, the amine salt can be dissolved in a suitable aprotic solvent and treated with a base and a suitable protecting group (PG) (e.g., PG = OSi, OBn, OMOM, etc.) to obtain 8. For example, the protection of primary alcohols with acid-stabilized silyl groups such as TBDMS or TBDPS is well known in the art.

[0116] Option 2

[0117]

[0118] Scheme 2 illustrates the preparation of the necessary substituted intermediate 3 (e.g., X = Br, Cl, I; R). 4 =F) as an alternative method. Those skilled in the art will recognize that a suitable disubstituted benzaldehyde 13(R) 4 The Horner-Wadsworth-Emmons condensation between 14 and N-protected amino-2-dimethoxyphosphoryl acetate 14 (e.g., PG=Z) can be used to prepare olefinic carbamate 15 (e.g., PG=Z). A subsequent chiral reduction under H2 with a suitable chiral auxiliary, such as with an Rh(I) transition metal catalyst, as well as well described in the art, yields N-protected 16. Exchange of the N-protecting group can be carried out under a variety of conditions recognizable to those skilled in the art to obtain 3. Subsequent transformations of compound 3 can be carried out as in Scheme 1 to provide compound 8 (e.g., X=Br, Cl, I; R...). 4 =F).

[0119] Option 3

[0120]

[0121] Scheme 3 describes the synthesis of type I' compounds. The formation of an imine from tetrahydroisoquinoline 8 can be accomplished under various oxidizing conditions recognizable to those skilled in the art, particularly the halogenation of a secondary amine followed by elimination with a suitable strong base to provide dihydroisoquinoline 9. A stereoselective Pierce reaction can be used to treat imine 9 with a suitable alkyl magnesium halide to obtain trans-tetrahydroisoquinoline 10. The relative configuration of tetrahydroisoquinoline 10 can be determined using appropriate NMR spectroscopy experiments, particularly 1D-NOESY. Subsequent N-acylation can be achieved using amide coupling techniques well known in the art, such as benzotriazol-1-yl-oxotripyrrolidinephosphonium hexafluorophosphate, in the presence of a mild, non-nucleophilic base, to prepare compound 11. Carbon-carbon bond formation using a suitably substituted compound 11 (e.g., X = Br, Cl, I, etc.) can be achieved under transition metal catalysis, such as using Pd, Pt, Ni, or Cu, with a suitable borate ester or trifluoroborane salt, as well known in the art. Alternatively, coupling with appropriately substituted olefins or alkynes under Heck- or Sonogashira-type conditions can be achieved as is known in the art. For example, palladium-mediated cross-coupling of 11 with appropriately substituted trifluoroboranuide can be achieved to obtain substituted cyclopropyl compounds of type 12. Additionally, palladium-mediated cross-coupling with appropriately substituted olefins of 11 can be achieved to obtain alkyl-substituted compounds of type 12. Those skilled in the art will recognize that deprotection of protected alcohol 12 (where PG = OSi, OBn, OMOM, etc.) can be carried out under a variety of conditions. For example, the silyl protecting group can be removed with tetrabutylammonium fluoride to obtain type I' chiral compounds.

[0122] In the following exemplary preparations and examples, the solvent is typically removed under reduced pressure (evaporation). In some methods, the indicated yield is a representative crude yield of the product separated by evaporation or filtration and used directly without further purification. The relative stereochemistry around the tetrahydroisoquinoline framework is determined by appropriate NMR spectroscopy, particularly 1D-NOESY. In the case of configurations around substituted cyclopropyl groups in side-chain substitutions, relative configurations are described without determining the absolute stereochemistry. In the case of additional chiral centers in side-chain substitutions, the absolute configuration has not yet been determined. In both cases, when separated by chiral chromatographic techniques, the diastereomer pair is designated as "Isomer 1" or "Isomer 2".

[0123] Preparation 1

[0124] 2-Bromo-D-phenylalanine methyl ester hydrochloride

[0125]

[0126] 2-Bromo-D-phenylalanine (22.4 g, 91.8 mmol) was dissolved in MeOH (459 mL). Acetyl chloride (65.3 mL, 917.7 mmol) was added at room temperature, and the resulting reaction mixture was stirred for 36 hours. The reaction mixture was concentrated under reduced pressure to give the title compound (27.2 g, >99% yield). ESMS (m / z) 258 / 260 [M-Cl, 79 Br / 81 Br].

[0127] Preparation 2

[0128] 2-Bromo-N-(methoxycarbonyl)-D-phenylalanine methyl ester

[0129]

[0130] 2-Bromo-D-phenylalanine methyl ester hydrochloride (27.2 g, 92.3 mmol) was dissolved in a two-phase mixture of DCM (923 mL) and water (185 mL). NaHCO3 (31 g, 369.4 mmol) and methyl chloroformate (7.9 mL, 101.6 mmol) were added at room temperature, and the resulting mixture was stirred for 2.5 hours. The mixture was diluted with water and extracted with DCM. The organic extract was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by rapid silica gel chromatography, eluting with a gradient of 10–75% EtOAc in hexane, and the solvent was evaporated from the desired chromatographic fraction to give the title compound (29.1 g, >99% yield). ESMS (m / z): 316 / 318 [M+H, 79 Br / 81 Br].

[0131] Preparation 3

[0132] (3R)-5-bromo-3,4-dihydro-1H-isoquinoline-2,3-dicarboxylic acid dimethyl ester

[0133]

[0134] A mixture of methyl 2-bromo-N-(methoxycarbonyl)-D-phenylalanine (29.1 g, 92.1 mmol) and paraformaldehyde (9.13 g, 101.3 mmol) was stirred for 7 hours in glacial acetic acid (115 mL, 2 mol) containing concentrated H₂SO₄ (38.4 mL, 719.9 mmol). The reaction mixture was partitioned between water and EtOAc, the layers were separated, and the aqueous layer was extracted with EtOAc. The combined organic extracts were dried over Na₂SO₄, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by rapid silica gel chromatography, eluting with a gradient of 5–40% EtOAc in hexane solution, and the solvent was evaporated to give the title compound (27.6 g, 91% yield). ESMS (m / z): 328 / 330 [M+H, 79 Br / 81 Br].

[0135] Preparation 4

[0136] (3R)-5-bromo-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid methyl ester hydrochloride

[0137]

[0138] Dimethyl (3R)-5-bromo-3,4-dihydro-1H-isoquinoline-2,3-dicarboxylate (27.6 g, 84 mmol) was dissolved in 5N HCl (330.6 mL, 1.7 mol), and the resulting mixture was heated under reflux for three days. The mixture was concentrated under reduced pressure to give a white solid. The solid was collected by filtration, washed with diethyl ether, and dried under vacuum at 40 °C overnight to give (3R)-5-bromo-1,2,3,4-tetrahydroisoquinoline-3-carboxylate (1:1) (20.8 g, 71.1 mmol). Acetyl chloride (50.6 mL, 711.0 mmol) was added to a mixture of (3R)-5-bromo-1,2,3,4-tetrahydroisoquinoline-3-carboxylate (1:1) (20.8 g, 71.1 mmol) in MeOH (474 ​​mL) at 0 °C. The mixture was warmed to room temperature and stirred for 36 hours. The mixture was concentrated under reduced pressure and dried to give the title compound (21.9 g, 85% yield). ESMS (m / z): 270 / 272 [M-Cl, 79 Br / 81 Br].

[0139] Preparation 5

[0140] 2-tert-butyl-3-methyl-(3R)-5-bromo-3,4-dihydro-1H-isoquinoline-2,3-dicarboxylic acid ester

[0141]

[0142] (3R)-5-bromo-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid methyl ester hydrochloride (21.0 g, 68.5 mmol) was dissolved in 1,4-dioxane (685 mL). A saturated aqueous solution of NaHCO3 (685 mL, 17.5 mol) and di-tert-butyl dicarbonate (29.9 g, 137.0 mmol) were added at room temperature, and the two-phase mixture was stirred at room temperature for 90 min. The mixture was extracted with EtOAc, the organic extract was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by rapid silica gel chromatography, eluting with a gradient of 5–50% EtOAc in hexane solution, and the solvent for the desired chromatographic fraction was evaporated to give the title compound (19.5 g, 77% yield). MS (m / z): 270 / 272 [M- t Boc+H, 79 Br / 81 Br].

[0143] Preparation of 6

[0144] (3R)-5-bromo-3-(hydroxymethyl)-3,4-dihydro-1H-isoquinoline-2-carboxylic acid tert-butyl ester

[0145]

[0146] A solution of 2M lithium borohydride (99.4 mL, 198.8 mmol) in THF and MeOH (10.1 mL, 248.5 mmol) was added to a solution of 2-tert-butyl-3-methyl-(3R)-5-bromo-3,4-dihydro-1H-isoquinoline-2,3-dicarboxylic acid ester (18.4 g, 49.7 mmol) in THF (497 mL), and the resulting mixture was stirred at room temperature for 40 min. The reaction was quenched with water and extracted with EtOAc. The organic extract was separated, dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by rapid silica gel chromatography, eluting with a gradient of 5-80% EtOAc in hexane. The solvent was evaporated from the desired chromatographic fraction, and the resulting residue was dried under high vacuum overnight to give the title compound as a white solid (19.1 g, >99% yield). ESMS (m / z): 286 / 288 [M- t Bu+H, 79 Br / 81 Br].

[0147] Preparation 7

[0148] [(3R)-5-bromo-1,2,3,4-tetrahydroisoquinoline-3-yl]methoxy-tert-butyl-dimethylsilane

[0149]

[0150] Trifluoroacetic acid (75.5 mL, 998.3 mmol) was added to a solution of (3R)-5-bromo-3-(hydroxymethyl)-3,4-dihydro-1H-isoquinoline-2-carboxylic acid tert-butyl ester (15.5 g, 45.3 mmol) in DCM (226 mL) at room temperature. The reaction mixture was stirred at room temperature for 30 min and concentrated under reduced pressure. The resulting residue was dried under vacuum to give [(3R)-5-bromo-1,2,3,4-tetrahydroisoquinoline-3-yl]methanol-2,2,2-trifluoroacetic acid as a wet solid. [(3R)-5-bromo-1,2,3,4-tetrahydroisoquinoline-3-yl]methanol-2,2,2-trifluoroacetic acid was dissolved in DCM (753 mL). 1H-imidazolium (51.3 g, 753 mmol), N,N-dimethyl-4-pyridinium (460 mg, 3.77 mmol), and tert-butyldimethylchlorosilane (13.6 g, 90.4 mmol) were added, and the resulting mixture was stirred overnight at room temperature. A saturated NH4Cl solution was added, and the mixture was extracted with DCM. The DCM layer was separated, dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure. The crude products were combined from a reaction substantially identical to that with (3R)-5-bromo-3-(hydroxymethyl)-3,4-dihydro-1H-isoquinoline-2-carboxylic acid tert-butyl ester (6.6 g, 19.4 mmol). The combined crude products were purified by rapid silica gel chromatography, eluting with a gradient of 5–40% EtOAc in hexane solution, and the title compound (14.3 g, 89% yield) was given after solvent evaporation of the desired chromatographic fraction. ESMS (m / z): 356 / 358 [M+H, 79 Br / 81 Br].

[0151] Preparation of 8

[0152] [(3R)-5-bromo-3,4-dihydroisoquinoline-3-yl]methoxy-tert-butyl-dimethylsilane

[0153]

[0154] [(3R)-5-bromo-1,2,3,4-tetrahydroisoquinoline-3-yl]methoxy-tert-butyl-dimethyl-silane (4.2 g, 11.8 mmol) was dissolved in diethyl ether (118 mL), and N-chlorosuccinimide (2.36 g, 17.7 mmol) was added. The resulting mixture was stirred at room temperature for 30 min and concentrated under reduced pressure. The resulting residue was dissolved in a solution of 5% KOH in MeOH (42.0 mL, 30.3 mmol) and stirred at room temperature for 30 min. The mixture was poured into water and extracted with DCM. The DCM extract was dried over Na2SO4 and filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by rapid silica gel chromatography, eluting with a gradient of 5-100% EtOAc in hexane solution to give the title compound (3.40 g, 82% yield). ESMS (m / z): 354 / 356 [M+H, 79 Br / 81 Br].

[0155] Alternatively, N-chlorosuccinimide (106.7 g, 790 mmol) was added to a solution of [(3R)-5-bromo-3,4-dihydroisoquinoline-3-yl]methoxy-tert-butyl-dimethyl-silane (220 g, 520 mmol) in THF (3.85 L), and stirred for 30 min at room temperature in a suitable container. The mixture was concentrated under reduced pressure, and the residue was dissolved in a methanol solution of 5% w / w KOH (2.2 L, 1.7 mol) while stirring for 30 min at room temperature. The mixture was added to water (3 L) and extracted three times with DCM (3 × 1 L). The combined organic extracts were dried over anhydrous MgSO4 and filtered, and the filtrate was concentrated under reduced pressure to give the title compound (210 g, >99% yield). ESMS (m / z): 354 / 356 [M+H, 79 Br / 81 Br].

[0156] Preparation 9

[0157] [(1S,3R)-5-bromo-1-methyl-1,2,3,4-tetrahydroisoquinoline-3-yl]methoxy-tert-butyl-dimethyl-silane

[0158]

[0159] [(3R)-5-bromo-3,4-dihydroisoquinoline-3-yl]methoxy-tert-butyl-dimethyl-silane (3.4 g, 9.6 mmol) was dissolved in diethyl ether (160 mL) and cooled to -78 °C in a dry ice-acetone bath. A solution of 3M magnesium methyl chloride in THF (26.9 mL, 80.6 mmol) was added dropwise. The mixture was slowly warmed to room temperature and stirred overnight. The reaction mixture was slowly quenched by adding saturated NH4Cl solution. The resulting mixture was extracted with DCM, dried over NaSO4, filtered, and the filtrate was concentrated under reduced pressure. The crude product was combined with the product from a reaction run of 1.7 mmol of [(3R)-5-bromo-3,4-dihydroisoquinoline-3-yl]methoxy-tert-butyl-dimethyl-silane in substantially the same manner. The combined residues were purified by rapid silica gel chromatography, eluting with a gradient of 5-65% EtOAc in hexane solution. The solvent for the desired chromatographic fraction was evaporated to give the title compound (3.78 g, >99% yield). ESMS (m / z): 370 / 372 [M+H, 79 Br / 81 Br].

[0160] The relative configuration of the compound [(1S,3R)-5-bromo-1-methyl-1,2,3,4-tetrahydroisoquinoline-3-yl]methoxy-tert-butyl-dimethyl-silane was determined by NMR spectroscopy using 1D-NOESY. Selective excitation of the methyl group at 1.30 ppm produced a NOE for Ha at 3.11 ppm. This NOE enhancement was only consistent with the configuration where the methyl group and Ha are on the same side of the ring (trans isomer), because in the cis isomer, the methyl proton is too far from Ha to exhibit a NOE. Since the absolute chemistry at position 3 is known to be R, the absolute chemistry at position 1 is deduced to be S.

[0161]

[0162] Preparation 10

[0163] 1-[(1S,3R)-5-bromo-3-[[tert-butyl(dimethyl)silyl]oxymethyl]-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]-2-(2,6-dichlorophenyl)ethyl ketone

[0164]

[0165] 2,6-Dichlorophenylacetic acid (3.7 g, 18 mmol) was combined with [(1S,3R)-5-bromo-1-methyl-1,2,3,4-tetrahydroisoquinoline-3-yl]methoxy-tert-butyl-dimethylsilane (5 g, 13.5 mmol), HATU (7.7 g, 20 mmol), and DIPEA (7.1 mL, 41 mmol) in a DCM (70 mL). The mixture was stirred at room temperature under a nitrogen atmosphere for 2 hours. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by rapid silica gel chromatography, eluting in a gradient of 0–25% EtOAc in hexane, followed by solvent evaporation of the desired chromatographic fraction to give the title compound (6.2 g, 81% yield). ESMS (m / z): 558 (M+1).

[0166] Preparation 11

[0167] trans-2-[(1S,3R)-3-[[tert-butyl(dimethyl)silyl]oxymethyl]-2-[2-(2,6-dichlorophenyl)acetyl]-1-methyl-3,4-dihydro-1H-isoquinoline-5-yl]cyclopropanecarboxylic acid ethyl ester

[0168]

[0169] In a microwave-safe vial, 1-[(1S,3R)-5-bromo-3-[[tert-butyl(dimethyl)silyl]oxymethyl]-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]-2-(2,6-dichlorophenyl)acetone (500 mg, 0.9 mmol) was combined with [trans-2-ethoxycarbonylcyclopropyl]-trifluoro-borohydride (300 mg, 1.3 mmol; see PCT / FR2013 / 053057), KH2PO4 (370 mg, 2.7 mmol), and 1,1'-bis(diphenylphosphino)ferrocene-palladium(II)dichloromethane complex (115 mg, 0.1 mmol) in 1,4-dioxane (8 mL, 93.2 mmol) and water (2 mL). The vial was capped, purged with nitrogen, and irradiated in a microwave at 120°C for 2 hours. The reaction solution was concentrated under reduced pressure, and the resulting residue was purified by rapid silica gel chromatography, eluting with a gradient of 0-25% EtOAc in hexane, and evaporating the solvent of the desired chromatographic fraction to give the title compound (245 mg, 46% yield) in the form of a mixture of trans isomers. ESMS (m / z): 590 (M+1).

[0170] Preparation 12

[0171] 1-[(1S,3R)-3-[[tert-butyl(dimethyl)silyl]oxymethyl]-5-[trans-2-(2-hydroxy-2-methyl-propyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]-2-(2,6-dichlorophenyl)ethyl ketone

[0172]

[0173] Ethyl trans-2-[(1S,3R)-3-[[tert-butyl(dimethyl)silyl]oxymethyl]-2-[2-(2,6-dichlorophenyl)acetyl]-1-methyl-3,4-dihydro-1H-isoquinoline-5-yl]cyclopropanecarboxylate (240 mg, 0.4 mmol) was stirred in THF (4 mL) and cooled to 0 °C. A solution of 3M magnesium methyl chloride in THF (1.75 mL, 5.25 mmol) was added dropwise over 5 minutes at 0 °C using a syringe. The resulting mixture was stirred under nitrogen for 1 hour, quenched with a saturated aqueous NH4Cl solution, and extracted with EtOAc. The organic layer was separated, dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to give the title compound, a mixture of trans isomers surrounding the cyclopropyl substituent, which was usable without further purification. ESMS (m / z): 576 (M+1).

[0174] Preparation 13

[0175] cis-2-[(1S,3R)-3-[[tert-butyl(dimethyl)silyl]oxymethyl]-2-[2-(2,6-dichlorophenyl)acetyl]-1-methyl-3,4-dihydro-1H-isoquinoline-5-yl]cyclopropanecarboxylic acid ethyl ester

[0176]

[0177] Essentially using the method described in Preparation 2, 1-[(1S,3R)-5-bromo-3-[[tert-butyl(dimethyl)silyl]oxymethyl]-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]-2-(2,6-dichlorophenyl)acetone (500 mg, 0.9 mmol) was combined with [cis-2-ethoxycarbonylcyclopropyl]-trifluoro-borohydride (WZ5-E16784-076-B, 300 mg, 1.3 mmol; see PCT / FR2013 / 053057) to give the title compound, a mixture of cis isomers (280 mg, 53% yield). ESMS (m / z): 590 (M+1).

[0178] Preparation 14

[0179] 1-[(1S,3R)-3-[[tert-butyl(dimethyl)silyl]oxymethyl]-5-[cis-2-(2-hydroxy-2-methyl-propyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]-2-(2,6-dichlorophenyl)acetone isomer 1

[0180] as well as

[0181] 1-[(1S,3R)-3-[[tert-butyl(dimethyl)silyl]oxymethyl]-5-[cis-2-(2-hydroxy-2-methyl-propyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]-2-(2,6-dichlorophenyl)acetone isomer 2

[0182]

[0183] Essentially using the method described in Preparation 3, a mixture of cis-2-[(1S,3R)-3-[[tert-butyl(dimethyl)silyl]oxymethyl]-2-[2-(2,6-dichlorophenyl)acetyl]-1-methyl-3,4-dihydro-1H-isoquinoline-5-yl]cyclopropanecarboxylic acid ethyl ester (280 mg, 0.5 mmol) and a solution of 3M methyl magnesium bromide in THF (2 mL, 6 mmol) was subsequently purified on silica gel by rapid chromatography using a 0-25% hexane solution gradient of EtOAc. Solvent evaporation of the desired chromatographic fraction yielded the title compound as the isolated cis isomer 1-[(1S,3R)-3-[[tert-butyl(dimethyl)silyl]oxymethyl]-2-[2-(2,6-dichlorophenyl)acetyl]-1-methyl-3,4-dihydro-1H-isoquinoline-5-yl]cyclopropanecarboxylic acid ethyl ester (280 mg, 0.5 mmol) and THF solution (2 mL, 6 mmol). R)-3-[[tert-butyl(dimethyl)silyl]oxymethyl]-5-[cis-2-(2-hydroxy-2-methyl-propyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]-2-(2,6-dichlorophenyl)acetone isomer 1 (110 mg, 38% yield) and 1-[(1S,3R)-3-[[tert-butyl(dimethyl)silyl]oxymethyl]-5-[cis-2-(2-hydroxy-2-methyl-propyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]-2-(2,6-dichlorophenyl)acetone isomer 2 (65 mg, 23% yield). ESMS (m / z): 576 (M+1) for each.

[0184] Preparation 15

[0185] 4-[(1S,3R)-3-[[tert-butyl(dimethyl)silyl]oxymethyl]-2-[2-(2-chloro-6-fluoro-phenyl)acetyl]-1-methyl-3,4-dihydro-1H-isoquinoline-5-yl]but-2-one

[0186]

[0187] 1-[(1S,3R)-5-bromo-3-[[tert-butyl(dimethyl)silyl]oxymethyl]-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]-2-(2-chloro-6-fluoro-phenyl)ethyl ketone (2 g, 3.7 mmol), tris(dibenzylacetone)dipalladium(0) (169 mg, 0.2 mmol), and di-tert-butyl-(1-phenylindol-2-yl)phosphine (187 mg, 0.55 mmol) were suspended in DMF (12.3 mL). N,N-methyldicyclohexylamine (0.9 mL, 4 mmol) and but-3-en-2-ol (0.45 mL, 5.2 mmol) were added, and the mixture was degassed by bubbling with nitrogen for 10 min. The reaction vessel was sealed and heated to 100 °C with stirring for 1.5 h. The reaction mixture was cooled to room temperature and diluted with EtOAc. The organic layer was separated, washed with a saturated aqueous sodium chloride solution, dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel rapid chromatography using a 5–30% hexane solution gradient of EtOAc, yielding the title compound (1.5 g, 75% yield) as a white foam after solvent evaporation of the desired chromatographic fraction. ESMS (m / z): 532 (M+1).

[0188] Preparation of 16

[0189] 1-((1S,3R)-3-(((tert-butyldimethylsilyl)oxy)methyl)-5-((R)-4,4-difluoro-3-methyl-3-((trimethylsilyl)oxy)butyl)-1-methyl-3,4-dihydroisoquinoline-2(1H)-yl)-2-(2-chloro-6-fluorophenyl)ethyl-1-one

[0190] as well as

[0191] 1-((1S,3R)-3-(((tert-butyldimethylsilyl)oxy)methyl)-5-((S)-4,4-difluoro-3-methyl-3-((trimethylsilyl)oxy)butyl)-1-methyl-3,4-dihydroisoquinoline-2(1H)-yl)-2-(2-chloro-6-fluorophenyl)ethyl-1-one

[0192]

[0193] 4-[(1S,3R)-3-[[tert-butyl(dimethyl)silyl]oxymethyl]-2-[2-(2-chloro-6-fluoro-phenyl)acetyl]-1-methyl-3,4-dihydro-1H-isoquinoline-5-yl]but-2-one (580 mg, 1.1 mmol) and 18-crown-6 (29 mg, 0.1 mmol) were dissolved in 1,2-dimethoxyethane (5.5 mL). (Difluoromethyl)trimethylsilane (0.30 mL, 2 mmol) was added, followed by CsF (17 mg, 0.1 mmol). The mixture was heated to 60 °C and stirred overnight. Further addition of (difluoromethyl)trimethylsilane (0.30 mL, 2 mmol), 18-crown-6 (29 mg, 0.1 mmol), and CsF (17 mg, 0.1 mmol) was made, and the reaction mixture was stirred at 60 °C for 4 hours. The mixture was cooled to room temperature, diluted with EtOAc, and the separated organic extract was washed with a saturated aqueous NaCl solution. The organic layer was dried over Na₂SO₄, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by rapid silica gel chromatography using a 0–10% hexane solution gradient of EtOAc, yielding a mixture of the title compounds as white foam (493 mg, 68% yield) after solvent evaporation of the desired chromatographic fraction. ESMS (m / z): 656 (M+1).

[0194] Preparation of 17

[0195] (Z)-2-(((benzyloxy)carbonyl)amino)-3-(2-bromo-3-fluorophenyl)methyl acrylate

[0196]

[0197] 2-(benzyloxycarbonylamino)-2-dimethoxyphosphoryl-acetate methyl ester (25.5 g, 76.9 mmol) was dissolved in DCM (250 mL), and the solution was cooled to 0 °C. 1,8-diazabicyclo[5.4.0]undec-7-ene (12 mL, 79.9 mmol) was added, and the mixture was stirred at 0 °C for 30 min. 2-Bromo-3-fluorobenzaldehyde (13.0 g, 64 mmol) was slowly added to the DCM (250 mL) solution over 20 min at 0 °C. The mixture was stirred at 0 °C for 2 h. The mixture was transferred to a separatory funnel and washed successively with saturated NH4Cl aqueous solution (200 mL) and saturated NaCl aqueous solution (200 mL). The organic layer was dried with Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to give a white solid. The solid was recrystallized from hot EtOAc and collected by filtration to give the title compound (25 g, 80% yield) as a white solid. ESMS(m / z):407(M+1).

[0198] Preparation of 18

[0199] Methyl (R)-2-(((benzyloxy)carbonyl)amino)-3-(2-bromo-3-fluorophenyl)propionate

[0200] BMD-E17046-003

[0201]

[0202] In a glove box, methyl (Z)-2-(((benzyloxy)carbonyl)amino)-3-(2-bromo-3-fluorophenyl)acrylate (26 g, 63.8 mmol) was added to an autoclave containing MeOH (300 mL). [((R)-tert-butylmethylphosphine)(di-tert-butylphosphine)amine](1,5-cyclooctadiene)rhodium(I)tetrafluoroborate (430 mg, 0.8 mmol) was added, the autoclave was sealed, and the mixture was removed from the glove box. The mixture was flushed with H2 gas for 15 seconds and pressurized to 100 psi H2. The resulting mixture was stirred at room temperature for 15 hours. The autoclave was vented, and the mixture was concentrated under reduced pressure to give the title compound as a white solid, usable without further purification (26.3 g, 98% yield). ESMS (m / z): 410 (M+1).

[0203] To obtain an analytically pure sample, a portion of the crude product (66 mg) was purified by silica gel rapid chromatography, eluting with a 15% EtOAc hexane solution. After evaporation of the solvent for the desired chromatographic fraction, a white solid product was obtained. Chiral HPLC analysis ( OJ-H column, 0.2% isopropanol / MeOH, 1 mL / min): Major enantiomer t R 2.981 minutes, minor enantiomer t R 3.557 minutes. Measurement >99% ee.

[0204] Preparation of 19

[0205] Methyl (R)-3-(2-bromo-3-fluorophenyl)-2-((methoxycarbonyl)amino)propionate

[0206]

[0207] Methyl (R)-2-(((benzyloxy)carbonyl)amino)-3-(2-bromo-3-fluorophenyl)propionate (21.25 g, 51.8 mmol) was dissolved in DCM (110 mL), and a solution of 33% HBr in acetic acid (28.1 mL, 155 mmol) was added. The resulting mixture was stirred at room temperature for 1 hour. Water (100 mL) and DCM (100 mL) were added, and the mixture was stirred vigorously for 15 minutes. The organic layer was separated and discarded. DCM (200 mL) and Na₂CO₃ (35.1 g, 331.5 mmol) were added to the aqueous layer. Methyl chloroformate (5.2 mL) was added to the mixture, and the resulting mixture was stirred at room temperature overnight. The mixture was extracted with DCM. The combined organic extracts were washed with a saturated aqueous solution of NaCl, dried over Na₂SO₄, filtered, and the filtrate was concentrated under reduced pressure to give the title compound as a white solid (13.6 g, 78% yield). ESMS(m / z):333(M+1).

[0208] Preparation 20

[0209] (R)-5-bromo-6-fluoro-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid hydrochloride

[0210]

[0211] Methyl (R)-3-(2-bromo-3-fluorophenyl)-2-((methoxycarbonyl)amino)propionate (17 g, 50.9 mmol) and paraformaldehyde (6.9 g, 76.4 mmol) were suspended in acetic acid (128 mL). The solution was cooled to 0 °C and concentrated H₂SO₄ (43 mL) was added. The resulting mixture was heated to 35 °C and stirred overnight. The reaction mixture was cooled to room temperature and poured into water (200 mL). The resulting suspension was extracted with EtOAc, and the combined organic extracts were washed with saturated NaCl, dried over Na₂SO₄, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was dissolved in concentrated HCl (231 mL), and the resulting mixture was heated to 120 °C overnight in a round-bottom flask equipped with a reflux condenser. The reaction mixture was cooled to room temperature and concentrated under reduced pressure at 50 °C. The resulting residue was dried under vacuum at 40 °C for 18 hours to give the title compound (15 g, 94% yield) as a colorless solid. ESMS(m / z):273(M+1).

[0212] Preparation 21

[0213] (R)-(5-bromo-6-fluoro-1,2,3,4-tetrahydroisoquinoline-3-yl)methanol

[0214]

[0215] (R)-5-bromo-6-fluoro-1,2,3,4-tetrahydroisoquinoline-3-carboxylate (9.95 g, 32.0 mmol) was suspended in THF (80 mL), and the mixture was cooled to 0 °C. A solution of 1 M borane-tetrahydrofuran complex (96 mL, 96 mmol) was slowly added. The mixture was heated to 60 °C and stirred overnight. The reaction mixture was cooled to 0 °C, and MeOH (20 mL) and saturated NH4Cl aqueous solution (40 mL) were added. The mixture was stirred at room temperature for 10 min. 5N HCl aqueous solution (60 mL) was added, and the resulting mixture was heated to 45 °C for 4 h. The mixture was cooled to 0 °C, and 21% (w / w) concentrated NH4OH aqueous solution was added. The mixture was warmed to room temperature. The resulting mixture was extracted with CHCl3:isopropanol in a 3:1 ratio. The combined organic extracts were washed with a saturated aqueous NaCl solution, dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to give the title compound (9.5 g, 86% yield) as a beige solid. ESMS (m / z): 259 (M+1).

[0216] Preparation 22

[0217] (R)-5-bromo-3-(((tert-butyldiphenylsilyl)oxy)methyl)-6-fluoro-1,2,3,4-tetrahydroisoquinoline

[0218]

[0219] (R)-(5-bromo-6-fluoro-1,2,3,4-tetrahydroisoquinoline-3-yl)methanol (12.6 g, 48.3 mmol) was dissolved in a mixture of DCM (185 mL) and DMF (146 mL). Imidazole (16.4 g, 241.5 mmol) and 4,4-dimethylaminopyridine (118 mg, 1 mmol) were added, followed by tert-butyldiphenylchlorosilane (13.8 mL, 53.1 mmol). The resulting mixture was stirred at room temperature for 1 hour. MeOH (100 mL) was added, and the mixture was stirred for another 1 hour. A saturated aqueous solution of NH4Cl (200 mL) was added, and the mixture was extracted with DCM. The combined organic extracts were washed with a saturated aqueous solution of sodium chloride, dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by rapid silica gel chromatography, eluting with a gradient of 2-50% EtOAc in hexane solution, followed by solvent evaporation of the desired chromatographic fraction to give the title compound in an oily form (10.5 g, 45% yield). ESMS (m / z): 498 M+1.

[0220] Preparation 23

[0221] (R)-5-bromo-3-(((tert-butyldiphenylsilyl)oxy)methyl)-6-fluoro-3,4-dihydroisoquinoline

[0222]

[0223] (R)-5-bromo-3-(((tert-butyldiphenylsilyl)oxy)methyl)-6-fluoro-1,2,3,4-tetrahydroisoquinoline (10.9 g, 21.9 mmol) was dissolved in THF (200 mL), and N-chlorosuccinimide (4.4 g, 32.9 mmol) was added. The resulting mixture was stirred at room temperature for 40 min and concentrated under reduced pressure. The resulting residue was dissolved in a solution of 0.75 M KOH in MeOH (100 mL) and stirred at room temperature for 30 min. The reaction mixture was diluted with water (50 mL) and extracted with DCM. The combined organic extracts were washed with a saturated aqueous NaCl solution, dried over Na2SO4, filtered, and the filtrate was concentrated. The resulting residue was purified by silica gel rapid chromatography, eluting with a gradient of 5-50% EtOAc in hexane solution, and evaporating the solvent of the desired chromatographic fraction to give the title compound (9.63 g, 88% yield) as a colorless oil. ESMS(m / z):496(M+1).

[0224] Preparation 24

[0225] (1S,3R)-5-bromo-3-(((tert-butyldiphenylsilyl)oxy)methyl)-6-fluoro-1-methyl-1,2,3,4-tetrahydroisoquinoline

[0226]

[0227] (R)-5-bromo-3-(((tert-butyldiphenylsilyl)oxy)methyl)-6-fluoro-3,4-dihydroisoquinoline (7.5 g, 15 mmol) was dissolved in tert-butyl methyl ether (190 mL) under nitrogen atmosphere, and the solution was cooled to -15 °C. A solution of 3 M magnesium methyl chloride in THF (20 mL, 60 mmol) was added dropwise with stirring, and the reaction mixture was warmed to room temperature and stirred for another 2 hours. The mixture was cooled to 0 °C, and MeOH (4 mL) and saturated NH4Cl aqueous solution (4 mL) were added sequentially. The resulting mixture was extracted with DCM. The combined organic extracts were washed with saturated sodium chloride aqueous solution, dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by rapid silica gel chromatography, eluting with a gradient of 2–5% tert-butyl methyl ether in DCM solution, and the solvent was evaporated after the desired chromatographic fraction to give the title compound (1.7 g, 23% yield) as a yellow oil. ESMS(m / z):512(M+1).

[0228] Preparation 25

[0229] 1-[(1S,3R)-5-bromo-3-[[tert-butyl(diphenyl)silyl]oxymethyl]-6-fluoro-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]-2-(2-chloro-6-fluoro-phenyl)ethyl ketone

[0230]

[0231] (1S,3R)-5-bromo-3-(((tert-butyldiphenylsilyl)oxy)methyl)-6-fluoro-1-methyl-1,2,3,4-tetrahydroisoquinoline (5.2 g, 10.1 mmol) was dissolved in DMF (101 mL). 2-(2-chloro-6-fluorophenyl)acetic acid (2.5 g, 13.2 mmol), 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazole[4,5-b]pyridine 3-oxide hexafluorophosphate (5 g, 13.2 mmol), and DIPEA (8.8 mL, 50.6 mmol) were added sequentially, and the resulting mixture was stirred overnight at room temperature. Water (200 mL) was added, and the mixture was extracted with EtOAc. The combined organic extracts were washed with saturated sodium chloride aqueous solution, dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by rapid silica gel chromatography, eluting with a gradient of 0-25% EtOAc in hexane solution, followed by solvent evaporation of the desired chromatographic fraction to give the title compound (6.3 g, 91% yield) as a white foam. ESMS (m / z): 682 (M+1).

[0232] Preparation 26

[0233] 1-[(1S,3R)-3-[[tert-butyl(diphenyl)silyl]oxymethyl]-6-fluoro-5-(3-hydroxy-3-methyl-but-1-ynyl)-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]-2-(2-chloro-6-fluoro-phenyl)ethyl ketone

[0234]

[0235] 1-[(1S,3R)-5-bromo-3-[[tert-butyl(diphenyl)silyl]oxymethyl]-6-fluoro-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]-2-(2-chloro-6-fluoro-phenyl)ethyl ketone (5.8 g, 8.5 mmol), tri-tert-butylphosphine(chloro)(crotonyl)palladium(II) (675 mg, 1.7 mmol), and cuprous salt of 2-thiophenecarboxylic acid (258 mg, 1.35 mmol) were added to a 500 mL round-bottom flask. The flask was evacuated and backfilled with nitrogen. DMF (60 mL), DIPEA (6 mL, 42.5 mmol), and 2-methylbut-3-yn-2-ol (4.1 mL, 42.2 mmol) were added. The reaction mixture was heated to 40 °C and stirred overnight. The mixture was cooled to room temperature, diluted with EtOAc, and the phases were separated. The organic layer was washed successively with water and saturated sodium chloride aqueous solution, dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by rapid silica gel chromatography, eluting with a gradient of 20–40% EtOAc in hexane solution, and evaporating the solvent of the desired chromatographic fraction to give the title compound (5.4 g, 93% yield) as a white foam. ESMS (m / z): 686 (M+1).

[0236] Preparation 27

[0237] 1-[(1S,3R)-3-[[tert-butyl(diphenyl)silyl]oxymethyl]-6-fluoro-5-(3-hydroxy-3-methyl-butyl)-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]-2-(2-chloro-6-fluoro-phenyl)ethyl ketone

[0238]

[0239] Purge the Parr shaker with nitrogen and add 5 wt% platinum sulfide / carbon (2.5 g). Purge the vessel again with nitrogen. Add EtOAc (50 mL). Add a solution of 1-[(1S,3R)-3-[[tert-butyl(diphenyl)silyl]oxymethyl]-6-fluoro-5-(3-hydroxy-3-methyl-but-1-ynyl)-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]-2-(2-chloro-6-fluoro-phenyl)acetone (5.4 g, 7.9 mmol) dissolved in EtOAc (50 mL). Purge the reaction vessel with H2 gas and pressurize to 60 psi H2. Shake the reaction mixture at room temperature for 3 hours. Add another 5 wt% carbon-supported platinum sulfide (1.21 g) suspended in EtOAc (30 mL), and pressurize the vessel again to 60 psi H2. Shake the reaction vessel again for 8 hours. The container was depressurized and purged with nitrogen. The resulting suspension was filtered and the filtrate was concentrated under reduced pressure. The resulting residue was purified by rapid silica gel chromatography, eluting with a gradient of 0-100% EtOAc in hexane solution (gradient 0-100%), and the title compound (5.33 g, 7.72 mmol) was given as a white foam after solvent evaporation of the desired chromatographic fraction. MS (m / z): 690.4 (M+1).

[0240] Preparation 28

[0241] trans-2-((1S,3R)-3-(((tert-butyldiphenylsilyl)oxy)methyl)-2-(2-(2-chloro-6-fluorophenyl)acetyl)-6-fluoro-1-

[0242] Ethyl methyl-1,2,3,4-tetrahydroisoquinoline-5-yl)cyclopropane-1-carboxylate

[0243]

[0244] 1-[(1S,3R)-5-bromo-3-[[tert-butyl(diphenyl)silyl]oxymethyl]-6-fluoro-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]-2-(2-chloro-6-fluoro-phenyl)acetone (665 mg, 1 mmol), racemic-[(trans-2-ethoxycarbonylcyclopropyl]-trifluoroborate (CAS#1612792-88-7; see PCT / FR2013 / 053057; December 12, 2013) (257 mg, 1.2 mmol), di(1-adamantyl)-n-butylphosphonium iodide (50 mg, 0.1 mmol), [(di(1-adamantyl)-butylphosphine)-2-(2'-amino-1,1'-biphenyl)]palladium methanesulfonate(II) (75 mg, 0.1 mmol) 1.9 mmol) and Cs2CO3 (631 mg, 1.9 mmol) were suspended in a mixture of toluene (10 mL) and water (1 mL). The mixture was degassed with nitrogen for 5 min, the vial was sealed, and heated to 100 °C overnight with stirring. The reaction mixture was cooled to room temperature and diluted with EtOAc. The resulting mixture was washed sequentially with water, saturated aqueous NaHCO3 solution, water, and saturated aqueous NaCl solution, and the layers were separated. The organic phase was dried over MgSO4 and filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by rapid silica gel chromatography, eluting with a gradient of 0-100% ethyl tert-butyl ether in hexane, and evaporating the solvent of the desired chromatographic fraction to give a mixture of the title compounds as a yellow solid (448 mg, 61% yield). ESMS (m / z): 716 (M+1).

[0245] Preparation 29

[0246] 1-[(1R,3R)-3-[[tert-butyl(diphenyl)silyl]oxymethyl]-6-fluoro-5-[(trans-2-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]-2-(2-chloro-6-fluoro-phenyl)ethyl ketone

[0247]

[0248] A mixture of trans-2-((1S,3R)-3-(((tert-butyldiphenylsilyl)oxy)methyl)-2-(2-(2-chloro-6-fluorophenyl)acetyl)-6-fluoro-1-methyl-1,2,3,4-tetrahydroisoquinoline-5-yl)cyclopropane-1-carboxylate isomer 1 and trans-2-((1S,3R)-3-(((tert-butyldiphenylsilyl)oxy)methyl)-2-(2-(2-chloro-6-fluorophenyl)acetyl)-6-fluoro-1-methyl-1,2,3,4-tetrahydroisoquinoline-5-yl)cyclopropane-1-carboxylate isomer 2 (440 mg, 0.6 mmol) was dissolved in THF (4 mL) and cooled to 0 °C. A solution of 1 M methyl magnesium bromide in THF (1.4 mL, 1.4 mmol) was slowly added over 10 minutes, and the resulting reaction mixture was heated to room temperature and stirred overnight. The reaction mixture was cooled to 0 °C, and another solution of 1 M methyl magnesium bromide in THF (1.4 mL, 1.4 mmol) was added. The reaction mixture was warmed to room temperature and stirred for 8 hours. The mixture was quenched with MeOH (130 μL) and stirred overnight at room temperature. The reaction mixture was diluted with EtOAc, and the resulting solution was washed successively with saturated aqueous NaHCO3 solution, water, and saturated aqueous NaCl solution. The layers were separated, the organic phase was dried over MgSO4 and filtered. The filtrate was concentrated under reduced pressure. The resulting residue was purified by rapid silica gel chromatography, eluting with a gradient of 0-100% methyl tert-butyl ether in hexane, and evaporating the solvent of the desired fraction to give a mixture of the title compounds as a yellow solid (245 mg, 42% yield). ESMS (m / z): 702 (M+1).

[0249] Preparation 30

[0250] 1-[(1S,3R)-5-bromo-3-[[tert-butyl(diphenyl)silyl]oxymethyl]-6-fluoro-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]-2-(2,6-dichlorophenyl)ethyl ketone

[0251]

[0252] (1S,3R)-5-bromo-3-(((tert-butyldiphenylsilyl)oxy)methyl)-6-fluoro-1-methyl-1,2,3,4-tetrahydroisoquinoline (500 mg, 1 mmol), 2-(2,6-dichlorophenyl)acetic acid (260 mg, 1.3 mmol), and 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazole[4,5-b]pyridinium 3-oxide hexafluorophosphate (482 mg, 1.3 mmol) were suspended in DMF (10 mL). DIPEA (0.85 mL, 4.9 mL) was added, and the mixture was stirred at room temperature for 72 hours. The reaction mixture was diluted with EtOAc and washed successively with water and saturated NaCl aqueous solution. The layers were separated, the organic layer was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by rapid silica gel chromatography, eluting with a gradient of 5–15% EtOAc in hexane solution, and evaporating the solvent of the desired fraction to give the title compound (680 mg, 0.97 mmol) as a white foam. ESMS (m / z): 698 (M+1).

[0253] Preparation 31

[0254] Ethyl trans-2-[(1S,3R)-3-[[tert-butyl(diphenyl)silyl]oxymethyl]-2-[2-(2,6-dichlorophenyl)acetyl]-6-fluoro-1-methyl-3,4-dihydro-1H-isoquinoline-5-yl]cyclopropanecarboxylate

[0255]

[0256] The following ingredients were added: 1-[(1S,3R)-5-bromo-3-[[tert-butyl(diphenyl)silyl]oxymethyl]-6-fluoro-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]-2-(2,6-dichlorophenyl)ethyl ketone (604 mg, 0.9 mmol), racemic-[(trans-2-ethoxycarbonylcyclopropyl]-trifluoroborate (CAS#1612792-88-7; see PCT / FR2013 / 053057; December 12, 2013) (228 mg, 1 mmol), di(1-adamantyl)-n-butylphosphonium iodide (44 mg, 0.1 mmol), [(di(1-adamantyl)-butylphosphine)-2-(2'-amino-1,1'-biphenyl)]palladium(II) methanesulfonate (66 mg, 0.1 mmol) and C S₂CO₃ (565 mg, 1.7 mmol) was suspended in a mixture of toluene (10 mL) and water (1 mL). The resulting mixture was bubbled with nitrogen for 5 minutes, the reaction was sealed, and the mixture was heated to 100 °C overnight with stirring. The reaction mixture was cooled to room temperature, diluted with EtOAc, and washed successively with water, saturated NaHCO₃ aqueous solution, water, and saturated NaCl aqueous solution. The layers were separated, the organic phase was dried over organic MgSO₄, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by rapid silica gel chromatography, eluting with a gradient of 0-100% methyl tert-butyl ether in hexane, and evaporating the solvent of the desired chromatographic fraction to give the title compound as a yellow solid, a mixture of trans-diastereomers (417 mg, 64% yield). ESMS (m / z): 732 (M+1).

[0257] Preparation of 32

[0258] 1-[(1S,3R)-3-[[tert-butyl(diphenyl)silyl]oxymethyl]-6-fluoro-5-[trans-2-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]-2-(2,6-dichlorophenyl)ethyl ketone

[0259]

[0260] A mixture (405 mg, 0.6 mmol) of trans-2-[(1S,3R)-3-[[tert-butyl(diphenyl)silyl]oxymethyl]-2-[2-(2,6-dichlorophenyl)acetyl]-6-fluoro-1-methyl-3,4-dihydro-1H-isoquinoline-5-yl]cyclopropanecarboxylate isomer 1 and trans-2-[(1S,3R)-3-[[tert-butyl(diphenyl)silyl]oxymethyl]-2-[2-(2,6-dichlorophenyl)acetyl]-6-fluoro-1-methyl-3,4-dihydro-1H-isoquinoline-5-yl]cyclopropanecarboxylate isomer 2 was dissolved in THF (3 mL), and the mixture was cooled to 0 °C. A solution of 1 M methylmagnesium bromide in THF (2.8 mL, 2.8 mmol) was slowly added over 10 minutes, and the reaction mixture was warmed to room temperature and stirred overnight. The reaction mixture was cooled to 0 °C and quenched with MeOH (220 μL). The reaction mixture was diluted with EtOAc and washed successively with water and saturated NaCl aqueous solution. The layers were separated, the organic phase was dried over MgSO4, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by rapid silica gel chromatography, eluting with a gradient of 0-100% methyl tert-butyl ether in hexane, and evaporating the solvent of the desired chromatographic fraction to give the title compound as a yellow solid, a mixture of trans-diastereomers (336 mg, 55% yield). ESMS (m / z): 718 (M+1).

[0261] Examples 1 and 2

[0262] 2-(2,6-Dichlorophenyl)-1-[(1S,3R)-3-(hydroxymethyl)-5-[trans-2-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]ethenone, isomer 1

[0263] as well as

[0264] 2-(2,6-Dichlorophenyl)-1-[(1S,3R)-3-(hydroxymethyl)-5-[trans-2-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]ethenone, isomer 2

[0265]

[0266] A solution of 1 M tetrabutylammonium formate in THF (0.8 mL) was added to a solution of 1-[(1S,3R)-3-[[tert-butyl(dimethyl)silyl]oxymethyl]-5-[(trans-2-(2-hydroxy-2-methyl-propyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]-2-(2,6-dichlorophenyl)acetone (170 mg, 0.3 mmol) in THF (2.5 mL), and the resulting mixture was stirred for 30 minutes. The reaction mixture was concentrated under reduced pressure. The resulting residue was purified by rapid silica gel chromatography, eluting with a gradient of 0-65% EtOAc in hexane solution, and further purified by rapid silica gel chromatography with 0-50% EtOAc solution. Gradient elution with DCM solution of EtOAc, followed by solvent evaporation of the desired chromatographic fraction, yielded the title compound 2-(2,6-dichlorophenyl)-1-[(1S,3R)-3-(hydroxymethyl)-5-[trans-2-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]ethenone isomer 2 (20 mg, 14% yield). The mixed fraction from another rapid chromatography was further purified by reversed-phase chromatography on 18C silica gel. Using a gradient of 5-95% acetonitrile solutions of water containing ammonium bicarbonate, solvent evaporation was performed on the desired chromatographic fraction to give the title compound 2-(2,6-dichlorophenyl)-1-[(1S,3R)-3-(hydroxymethyl)-5-[trans-2-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]ethenone, isomer 1 (26 mg, 18% yield). ESMS (m / z): 462 (M+1) for each. 1 H nmr (400MHz, dmso-d6) isomer 1: δ 0.66-1.01 (m, 4H), 1.08-1.27 (m, 6H), 1.40-1.89 (m, 1H), 2.00-2.08 (m, 1H), 2.66-3.04 (m, 2H), 3.20-3.32 (m, 2H), 3.63-3.75 (m, 1H), 4.09-4.31 (m, 3H), 4.38-4.50 (m, 1H), 4.98-5.25 (m, 2H), 6.88-7.16 (m, 3H), 7.33 (t, 1H), 7.48 (d, 2H). 1H nmr (400MHz, dmso-d6) isomer 2: δ 0.66-1.01 (m, 4H), 1.06-1.31 (m, 5H), 1.40-1.89 (m, 2H), 1.99-2.05 (m, 1H), 2.67-3.00 (m, 2H), 3.24-3.32 (m, 2H), 3.66-3.77 (m, 1H), 4.09-4.01 (m, 3H), 4.40-4.49 (m, 1H), 4.98-5.25 (m, 2H), 6.92-7.15 (m, 3H), 7.34 (t, 1H), 7.48 (d, 2H).

[0267] Examples 3 and 4

[0268] 2-(2,6-Dichlorophenyl)-1-[(1S,3R)-3-(hydroxymethyl)-5-[cis-2-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]ethenone, isomer 1

[0269] as well as

[0270] 2-(2,6-Dichlorophenyl)-1-[(1S,3R)-3-(hydroxymethyl)-5-[cis-2-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]ethenone, isomer 2

[0271]

[0272] Using essentially the methods described in Examples 1 and 2, 1-[(1S,3R)-3-[[tert-butyl(dimethyl)silyl]oxymethyl]-5-[cis-2-(2-hydroxy-2-methyl-propyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]-2-(2,6-dichlorophenyl)ethenone isomer 1 (100 mg, 173 mmol) was used to give the title compound 2-(2,6-dichlorophenyl)-1-[(1S,3R)-3-(hydroxymethyl)-5-[cis-2-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]ethenone isomer 1 (61 mg, 76% yield). ESMS (m / z): 462 (M+1). 1Hnmr (400MHz, dmso-d6) isomer 1: δ 0.55 (s, 1H), 0.65 (s, 2H), 0.94-1.00 (m, 2H), 1.07-1.22 (m, 7H), 1.40-1.49 (m, 2H), 1.98-2.07 (m, 1H), 2.60-3.15 (m, 2H), 3.20-3.29 (m, 1H), 3.53-3.65 (m, 1H), 4.09-4.34 (m, 2H), 4.37-4.48 (m, 1H), 4.98-5.34 (m, 2H), 7.00-7.22 (m, 3H), 7.30-7.37 (m, 1H), 7.45-7.51 (m, 2H).

[0273] Using essentially the methods described in Examples 1 and 2, 1-[(1S,3R)-3-[[tert-butyl(dimethyl)silyl]oxymethyl]-5-[cis-2-(2-hydroxy-2-methyl-propyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]-2-(2,6-dichlorophenyl)ethenone isomer 2 (60 mg, 104 mmol) was used to give the title compound 2-(2,6-dichlorophenyl)-1-[(1S,3R)-3-(hydroxymethyl)-5-[cis-2-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]ethenone isomer 2 (42 mg, 87% yield). ESMS (m / z) for each: 462 (M+1). 1 H nmr (400MHz, dmso-d6) isomer 1: δ 0.81 (s, 3H), 0.83-0.94 (m, 4H), 1.14-1.26 (m, 5H), 1.28-1.37 (m, 1H), 1.50 (d, 1H), 2.09-2.18 (m, 1H), 2.66-3.05 (m, 2H), 3.27-3.32 (m, 1H), 3.56-3.67 (m, 1H), 4.08-4.32 (m, 2H), 4.39-4.48 (m, 1H), 4.96-5.26 (m, 2H), 7.00-7.16 (m, 3H), 7.33 (t, 1H), 7.48 (d, 2H).

[0274] Examples 5 and 6

[0275] 2-(2,6-Dichlorophenyl)-1-[(1S,3R)-5-[4,4-difluoro-3-hydroxy-3-methyl-butyl]-3-(hydroxymethyl)-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]ethenone, isomer 1

[0276] as well as

[0277] 2-(2,6-Dichlorophenyl)-1-[(1S,3R)-5-[4,4-difluoro-3-hydroxy-3-methyl-butyl]-3-(hydroxymethyl)-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]ethenone, isomer 2

[0278]

[0279] A mixture of 2-(2,6-dichlorophenyl)-1-[(1S,3R)-5-[(3R)-4,4-difluoro-3-hydroxy-3-methyl-butyl]-3-(hydroxymethyl)-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]acetone and 2-(2,6-dichlorophenyl)-1-[(1S,3R)-5-[(3S)-4,4-difluoro-3-hydroxy-3-methyl-butyl]-3-(hydroxymethyl)-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]acetone (493 mg, 0.75 mmol) was dissolved in THF (12 mL). A solution of 1 M tetrabutylammonium fluoride in THF (3 mL, 3 mmol) was added, and the resulting mixture was stirred at room temperature for 1 hour. The mixture was diluted with EtOAc and washed with a saturated aqueous NaCl solution. The organic layer was separated, dried over Na₂SO₄, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel rapid chromatography using a hexane solution gradient of 0–75% EtOAc, yielding a white, foamy title mixture (342 mg, 97% yield) after evaporation of the desired chromatographic fraction. ESMS (m / z): 470 (M+1).

[0280] Through chiral SFC ( The two diastereomers were further purified and separated using an OD-H column (21×250 mm). The diastereomers were eluted with methanol:CO2 (15:85) at a flow rate of 80 mL / min and a temperature of 40 °C. After evaporation of the solvent for the desired chromatographic fraction, isomer 1 (156 mg, 46% yield) and isomer 2 (141 mg, 41% yield) were obtained.

[0281] Isomer 1: ESMS (m / z): 470 (M+H). Analytical SFC t R 1.618 minutes OD-H column, 4×150mm, 15% methanol / CO2, 5mL / min, 225nm.

[0282] Isomer 2: ESMS (m / z): 470 (M+H). Analytical SFC t R 2.213 minutes OD-H column, 4×150mm, 15% methanol / CO2, 5mL / min, 225nm.

[0283] Example 7

[0284] 2-(2-chloro-6-fluoro-phenyl)-1-[(1S,3R)-6-fluoro-3-(hydroxymethyl)-5-(3-hydroxy-3-methyl-butyl)-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl] ethyl ketone

[0285]

[0286] 1-((1S,3R)-3-(((tert-butyldiphenylsilyl)oxy)methyl)-6-fluoro-5-(3-hydroxy-3-methylbutyl)-1-methyl-3,4-dihydroisoquinoline-2(1H)-yl)-2-(2-chloro-6-fluorophenyl)ethyl-1-one (5.3 g, 7.7 mmol) was dissolved in THF (129 mL), and a solution of 1 M tetrabutylammonium fluoride in THF (23 mL, 23 mmol) was added. The mixture was stirred at room temperature for 1 hour. The mixture was diluted with EtOAc, washed with a saturated aqueous solution of NaCl, and the layers were separated. The organic extract was dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by rapid silica gel chromatography, eluting with a gradient of 50-100% EtOAc in hexane, and evaporating the solvent of the desired chromatographic fraction to give the title compound (2.75 g, 79% yield) as a white solid. ESMS(m / z):452(M+1).

[0287] Examples 8 and 9

[0288] 2-(2-chloro-6-fluoro-phenyl)-1-[(1R,3R)-6-fluoro-3-(hydroxymethyl)-5-[trans-2-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]ethenone, isomer 1

[0289] as well as

[0290] 2-(2-chloro-6-fluoro-phenyl)-1-[(1R,3R)-6-fluoro-3-(hydroxymethyl)-5-[trans-2-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]ethenone, isomer 2

[0291]

[0292] A mixture (1.1 g, 1.5 mmol) of 1-[(1R,3R)-3-[[tert-butyl(diphenyl)silyl]oxymethyl]-6-fluoro-5-[trans-2-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]-2-(2-chloro-6-fluoro-phenyl) ketone isomer 1 and 1-[(1R,3R)-3-[[tert-butyl(diphenyl)silyl]oxymethyl]-6-fluoro-5-[trans-2-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]-2-(2-chloro-6-fluoro-phenyl) ketone isomer 2] was dissolved in THF (10 mL), and the mixture was cooled to 0 °C. A solution of 1 M tetrabutylammonium fluoride in THF (3 mL, 3 mmol) was added dropwise, and the reaction mixture was stirred and warmed to room temperature overnight. The reaction mixture was diluted with EtOAc and washed successively with water and saturated NaCl aqueous solution. The resulting layer was separated, the organic layer was dried over MgSO4, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by rapid silica gel chromatography, eluting with a gradient of 0-100% methyl tert-butyl ether in hexane, and evaporating the solvent of the desired chromatographic fraction to give a mixture of the title compounds as an amber oil (490 mg, 69% yield). ESMS (m / z): 464 (M+1).

[0293] The two diastereomers were further purified and separated by chiral SFC. Cellulose-2 column, 21×250 mm, 20% isopropanol / CO2, 80 mL / min, 40 °C), after solvent evaporation for the desired chromatographic fraction, yielded a white solid 2-(2-chloro-6-fluoro-phenyl)-1-[(1R,3R)-6-fluoro-3-(hydroxymethyl)-5-[trans-2-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]ethenone, isomer 1 (136 mg, 28% yield; analytical HPLC t) R : 2.769 min) and 2-(2-chloro-6-fluoro-phenyl)-1-[(1R,3R)-6-fluoro-3-(hydroxymethyl)-5-[trans-2-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]ethenone, isomer 2 (121 mg, 25% yield; analytical HPLC t R (3.368 minutes).

[0294] Isomer 1: ESMS(m / z): 464(M+H). 1H NMR (400MHz, CDCl3): δ0.94-1.17(m,2H),1.17-1.25(m,3H),1.25-1.44(m,4H),1.60(s,3H),1.80-1.90(m,1H),2.07-2.37(m,1H),2.91- 3.23(m,3H),3.47-3.57(m,1H),3.92(t,J=16.1Hz,1H),4.05(s,2H),4.42-4.66(m,1H),5.04(q,J=7.0Hz,0.5H),5.21(q,J=7.0Hz,0.5H). 6.83-7.09(m,3H),7.19-7.26(m,2H). Analytical SCF t R 2.769 minutes ( Cellulose-2 column, 4×150mm, 20% isopropanol / CO2, 5mL / min, 225nm.

[0295] Isomer 2: ESMS(m / z): 464(M+H). 1 H NMR (400MHz, CDCl3): δ0.71-0.81(m,1H),1.22(s,3H),1.24(s,3H),1.33-1.44 (m,4H),1.48-1.59(m,1H),1.59-1.69(m,2H),1.69-1.81(m,1H),2.87-3.08(m ,1H),3.08-3.31(m,1H),3.47-3.57(m,1H),3.70-3.95(m,1H),3.97-4.11(m,2 H),4.43-4.60(m,1H),5.01-5.26(m,1H),6.84-7.08(m,3H),7.20-7.26(m,2H). Analytical SFC t R 3.368 minutes ( Cellulose-2 column, 4×150mm, 20% isopropanol / CO2, 5mL / min, 225nm.

[0296] Examples 10 and 11

[0297] 2-(2,6-Dichlorophenyl)-1-[(1S,3R)-6-fluoro-3-(hydroxymethyl)-5-[trans-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]ethenone, isomer 1

[0298] as well as

[0299] 2-(2,6-Dichlorophenyl)-1-[(1S,3R)-6-fluoro-3-(hydroxymethyl)-5-[trans-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]ethenone, isomer 2

[0300]

[0301] A mixture (333 mg, 0.5 mmol) of 1-[(1S,3R)-3-[[tert-butyl(diphenyl)silyl]oxymethyl]-6-fluoro-5-[trans-2-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]-2-(2,6-dichlorophenyl)ethyl ketone isomer 1 and 1-[(1S,3R)-3-[[tert-butyl(diphenyl)silyl]oxymethyl]-6-fluoro-5-[trans-2-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]-2-(2,6-dichlorophenyl)ethyl ketone isomer 2] was dissolved in THF (4.6 mL), and the mixture was cooled to 0 °C. A solution of 1 M tetrabutylammonium fluoride was added dropwise to a solution of THF (0.50 mL, 0.50 mmol), and the resulting reaction mixture was heated to room temperature and stirred overnight. The reaction mixture was diluted with EtOAc and washed successively with water and saturated NaCl aqueous solution. The layers were separated, the organic phase was dried over MgSO4, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by rapid silica gel chromatography, eluting with a gradient of 0-100% methyl tert-butyl ether in hexane, and evaporating the solvent of the desired chromatographic fraction to give a mixture of the title compounds as an amber oil (211 mg, 72% yield). ESMS (m / z): 480 (M+1).

[0302] The two diastereomers were further purified and separated by chiral SFC. Cellulose-2 column, 21 × 250 mm, 20% IPA / CO2, 80 mL / min, 40 °C), after solvent evaporation of the desired chromatographic fraction, yielded the title compound 2-(2,6-dichlorophenyl)-1-[(1S,3R)-6-fluoro-3-(hydroxymethyl)-5-[trans-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]ethenone as a white solid, isomer 1 (41 mg, 21% yield; analytical HPLC t). R: 3.095 min) and 2-(2,6-dichlorophenyl)-1-[(1S,3R)-6-fluoro-3-(hydroxymethyl)-5-[trans-(1-hydroxy-1-methyl-ethyl)cyclopropyl]-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl]ethenone, isomer 2 (47 mg, 24% yield; analytical HPLC t R (3.793 minutes).

[0303] Isomer 1: ESMS(m / z): 480(M+H). 1 ¹H NMR (400MHz, CDCl₃): δ 0.92–1.07 (m, 2H), 1.20–1.27 (m, 5H), 1.60–1.66 (d, J = 6.6 Hz, 1H), 1.80–1.90 (m, 1H). 2.93–3.23 (m, 2H), 3.47–3.60 (m, 1H), 3.81–4.00 (m, 1H), 4.23 (s, 2H), 4.44–4.70 (m, 1H), 5.06 (q, J = 6.7 Hz, 0.5H), 5.23 (q, J = 6.7 Hz, 0.5H), 6.84–7.04 (m, 2H), 7.16–7.23 (m, 1H), 7.36 (d, J = 8.1 Hz, 2H). Analytical SFC t R 3.095 minutes ( Cellulose-2 column, 4.6 × 150 mm, 20% IPA / CO2, 5 mL / min, 225 nm.

[0304] Isomer 2: ESMS(m / z): 480(M+H). 1 H NMR (400MHz, CDCl3): δ0.70-0.83(m,1H),1.09-1.18(m,1H),1.23(d,J=6.1Hz, 1H),1.32(s,3H),1.35-1.46(m,4H),1.64(d,J=6.7Hz,2H),1.69-1.82(m,1H), 2.90-3.10(m,1H),3.16-3.37(m,1H),3.50-3.61(m,1H),3.70-4.00(m,1H),4. 23 (s, 2H), 4.48-4.63 (m, 1H), 5.09 (q, J = 6.7Hz, 0.6H), 5.24 (q, J = 6.7Hz, 0.4H). 6.85–7.05 (m, 2H), 7.15–7.23 (m, 1H), 7.36 (d, J = 8.0 Hz, 2H). Analytical SFC t R 3.793 minutes ( Cellulose-2 column, 4.6 × 150 mm, 20% IPA / CO2, 5 mL / min, 225 nm.

[0305] Human D1 receptor PAM assay

[0306] The PAM activity of the compounds of this invention can be measured essentially as described in Svensson et al., An Allosteric Potentiator of the Dopamine D1 Receptor Increases Locomotor Activity in Human D1 Knock-in Mices without Casusing Stereotypy or Tachyphylaxis, J. Pharmacol. Exp. Ther. (2017) 360: 117-128.

[0307] More specifically, HEK293 cells stably expressing the human D1 receptor (accession number NM_000794) were generated via gene transduction using the pBABE-bleo retroviral vector and treated with Zeocin. TM (InvivoGen) selection. Use TrypLE when the convergence is around 80%. TM Cells were harvested via Express (Gibco), suspended in FBS with 8% DMSO, and stored in liquid nitrogen. On the day of assay, cells were thawed and resuspended in STIM buffer (Hankes balanced salt solution supplemented with 0.1% BSA, 20 mM HEPES, 500 μM IBMX, and 100 μM ascorbic acid).

[0308] The test compound was serially diluted (1:2) with DMSO into an assay plate (ProxiPlate-384Plus, PerkinElmer) using an acoustic dispensing (Labcyte) method to provide 20 concentrations for full response curves. The test compound (80 nL) was added to 5 μL of STIM buffer containing 2000 cells and 5 μL of 2X dopamine solution in STIM buffer, which will generate EC. 20 The level response was 24 nM in the stock solution or 12 nM finally, with a final DMSO concentration of 0.8% in the wells. The plates were incubated at room temperature for a total reaction time of 60 minutes.

[0309] According to the manufacturer's instructions, use Detection (Cisbio) to quantify cAMP production. Typically, a cavitary compound containing anti-cAMP (5 μL) and a D2-conjugate (from...) are used. Add 5 μL of lysis buffer (from the kit) to the wells, incubate the plate for another 60 to 90 minutes, and then use EnVision... TM Time-resolved fluorescence was detected using a plate reader (PerkinElmer). Fluorescence data were converted to cAMP concentrations using a cAMP standard curve and analyzed using a 4-parameter nonlinear logistic equation (GenedataScreener, version 13.0.5 - standard). For synergist mode concentration-response curves, results are expressed as EC50 values ​​for dopamine alone. 20 The percentage window between the response at a given concentration (normalized to 0%) and the maximum response to dopamine (defined as the response to 5 μM dopamine at the final concentration, normalized to 100%).

[0310] The absolute EC was calculated based on the maximum and minimum responses of the control agonist (dopamine). 50 Value. % enhancement (% top / % top) is determined by fitting the top of the concentration response curve. Absolute EC50 values ​​for certain example compounds. 50 The highest values ​​for % are shown in Table 1 below:

[0311] Table 1:

[0312]

[0313] The absolute EC values ​​of the above example compounds in Table 1 50 The values ​​indicate enhanced signaling at the human D1 receptor in response to dopamine and demonstrate the activity of the compound of claim 1 as a positive allosteric modulator of the human dopamine D1 receptor. The example compounds 1-6 and 8-11 of the present invention represent chiral compounds and, as described in the examples herein, have been prepared and tested as individual stereoisomers. See Examples 1-6, 8-11 above and Table 1. Combinatorial data of individual stereoisomers (absolute EC50 values ​​of Examples 1-6, 8-11) 50 The range (from 5.6 nM to 86.7 nM) demonstrates that each individual stereoisomer represents an embodiment of the D1 orthoallosteric modifier of the present invention. The characterization and determination of the absolute stereochemistry of the individual stereoisomers of the examples provided herein are within the skill of those skilled in the art, and the methods used for such determinations are well known in the medicinal chemistry literature (see, for example, Chiral Analysis (Second Edition) Advances in Spectroscopy, Chromatography and Emerging Methods, 2018). For example, absolute configuration is typically determined by NMR based on CDA: diastereomeric derivatives involving covalent binding between a chiral adjuvant and an enantiomeric substrate adopt a preferred conformation, which can be predicted based on differential shielding caused by the aromatic ring bound to the chiral identifying agent.

[0314] Generation of human D1 receptor knock-in mice

[0315] Transgenic mice in which the mouse dopamine 1 (D1) receptor is replaced by its human counterpart can be generated using standard techniques (see generally Svensson et al., J. Pharmacol. Exp. Ther. (2017) 360: 117–128). For example, a mouse genome fragment is subcloned from an RP23 bacterial artificial chromosome library and then cloned into a PGK-neo targeting vector. The mouse open reading frame is replaced with the human D1 receptor open reading frame in exon 2. The neo selection marker flanking exon 2 is a frt site for subsequent removal. Exon 2 also has a loxP selection site flanking it, allowing selection to generate D1 knockout mice by hybridization with mice expressing the cre nuclease gene.

[0316] C57BL / 6N embryonic stem cell line B6-3 was cultured in a solution containing 20% ​​FBS and 2×10⁻⁶ oz. 6 Mouse embryonic fibroblasts were grown on a feeder layer containing high-glucose DMEM containing leukemia inhibitory factor and mitosis inactivation. 10 million embryonic stem cells were electroporated with 30 μg of linearized vector DNA and selected using G418 (200 μg / ml). Clones were isolated and analyzed by Southern blotting.

[0317] Clones containing the intended-size insert were inserted into blastocysts, and the resulting mice were genotyped by PCR. Male chimeras were crossed with female chimeras containing the flp nuclease gene to eliminate the selection marker. Offspring containing the marker-free human D1 receptor were identified by PCR. Male heterozygotes were mated with female C57BL / 6 mice. Male and female offspring containing the human D1 receptor were mated, and homozygotes were identified by PCR. Homozygotes were found to exhibit normal behavior and reproduction, and the population remained homozygous in subsequent generations.

[0318] Basic (habitual) motor activity

[0319] The in vivo efficacy of the compounds of this invention can be demonstrated by acting through the use of D1 receptors with kinetic activity in mice. Kinetic activity was measured by tracking mouse movement using an automated system. Mouse kinetic behavior was monitored in transparent plastic shoebox cages measuring 45×25×20cm, with 1cm deep wooden blocks used as absorbent bedding and a ventilated, filtered plastic top. The cages were placed within a rectangular frame containing a grid of 12 phototube beams (KinderScientific, Poway, CA) in an 8×4 configuration, located 2.5cm from the cage floor, for detecting body movement (walking) and recording it by a computer.

[0320] Male human D1 receptor knock-in mice were placed in a chamber and allowed to acclimatize for 60 minutes. During the acclimatization period, the mice exhibited the expected decrease in movement over time. After administration of the compound of the present invention, an increase in animal movement was observed in a dose-dependent manner.

[0321] Mice were randomly assigned to treatment groups. In the dose-response study, each mouse was placed in a 60-minute acclimatization box. The test compound, in a 20% hydroxypropyl-β-cyclodextrin carrier, was then orally administered to the mice at a dose volume of 10 mL / kg. After administration, the mice were returned to the LMA box, and the total number of steps taken by each mouse was recorded every 10 minutes during the 60-minute measurement period. Statistical analysis was performed using one-way ANOVA, followed by post-hoc analysis using Dunnett's test of comparisons.

[0322] The compounds in Example 7 were determined in a dose-dependent manner as described above, and it was found that they increased basal motor function in a dose-dependent manner (Table 2 below).

[0323] Table 2:

[0324]

[0325] The basic motor activity data for Example 7 shown in Table 2 illustrate that the compounds of the present invention, particularly Example 7, are effective in motor activation in environmentally adapted animals. This activity is believed to result from allosteric enhancement and central activation of the D1 receptor (see, for example, Svensson et al., J. Pharmacol. Exp. Ther. (2017) 360: 117–128). The data for Example 7 provided in Table 2 illustrate the pharmacologically favorable in vivo efficacy of the compounds of the present invention in enhancing endogenous dopamine-mediated responses. The data for Example 7 provided in Table 2 further illustrate the pharmacologically favorable oral bioavailability of Example 7 and compounds having Formula I.

[0326] Plasma and brain levels:

[0327] 2-(2-chloro-6-fluoro-phenyl)-1-[(1S,3R)-6-fluoro-3-(hydroxymethyl)-5-(3-hydroxy-3-methyl-butyl)-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]ethinone of Example 7 was orally administered to male mice at doses ranging from 3 mg / kg to 60 mg / kg under feeding conditions, and plasma and brain concentrations were measured 1 hour after administration. The unbound fraction of the compound was determined in vitro as previously described (Zamek-Gliszczynski Mj et al., Validation of 96-well equilibrium dialysis with non-radiolabeled drug for definitive measurement of protein binding and application to clinical development of highly-bound drugs. J. Pharm. Sci., (2011) 100: 2498-2507). As previously described, the ratio of unbound brain concentration (Cu, brain) to unbound plasma concentration (Cu, plasma) was measured (Kpuu) (Raub Tj et al., Brain Exposure of Two Selective Dual CDK4 and CDK6 Inhibitors and the Antitumor Activity of CDK4 and CDK6 Inhibition in Combination with Temozolomide in an Intracranial Glioblastoma Xenograft, Drug Metab. Dispos. (2015) 43:1360-71). The data for Example 7 given in Table 3 below are averages from three animals at each dose. “Con.” indicates concentration.

[0328] Table 3:

[0329]

[0330] The compounds of the present invention, such as Example 7, 2-(2-chloro-6-fluoro-phenyl)-1-[(1S,3R)-6-fluoro-3-(hydroxymethyl)-5-(3-hydroxy-3-methyl-butyl)-1-methyl-3,4-dihydro-1H-isoquinolin-2-yl]ethenone, exhibit a favorable combination of pharmacological properties, such as enhanced human D1 receptor signaling in response to dopamine, high oral in vivo availability, central nervous system availability, and in vivo efficacy in motor activation in acclimated animals. For example, Example 7 demonstrated enhanced human D1 receptor signaling in response to dopamine (10.2 ± 1.40 nM (n = 15)) and significant in vivo efficacy in motor activation in acclimated human D1 receptor knock-in mice when administered orally at 10, 30, and 60 mg / kg PO, illustrating the compound's favorable oral bioavailability. Furthermore, Example 7 was generally well-tolerated when administered to normal rats over a wide dose range and showed a favorable lack of toxicity in this in vivo experiment. Therefore, Example 7 demonstrates a favorable combination of pharmacological properties that support its potential use as a therapeutic agent for oral administration to enhance dopamine D1 receptor function, and for the treatment of Parkinson's disease, schizophrenia, ADHD, and / or Alzheimer's disease.

[0331] via CYP3A4(fm) CYP3A4 Determination of the fraction of compounds eliminated by metabolism

[0332] The fraction of total clearance of a drug via oxidative metabolism through cytochrome P450 (CYP) is an important indicator of potential victim-drug interaction-mediated side effects. (See generally Ogu, Chris C and Maxa, Jan L, Drug interactions due to cytochrome P450, BUMC Proceedings 2000; 13:421-423). A higher fraction of total clearance for a given drug via CYP oxidative pathways, particularly via a single CYP (e.g., CYP3A4), indicates a greater likelihood that the drug will be sensitive to undesirable victim-drug interactions when used for treatment. To determine fm... CYP3A4 (Fractions of CYP3A4 metabolism) were first determined in human hepatocytes, specifically the fraction of P450-mediated oxidation (fm). CYP Then, CYP3A4-mediated oxidation (fm) was determined using recombinant CYP phenotyping (RCP). CYP3A4_RCP The relative contribution of fm to other P450 values. Calculate fm using the following equation. CYP3A4 :

[0333] fm CYP3A4 =fm CYP x fm CYP3A4_RCP

[0334] Cytochrome P450 (CYP)-mediated oxidative metabolism (fm) in human hepatocytes CYP Determination of fractions

[0335] In the presence and absence of the pan-P450 inhibitor 1-aminobenzotriazole (ABT), the relative contribution of P450 to the overall metabolism of the compound is determined by drug clearance in assays using human hepatocytes. Essentially, the intrinsic clearance in refrigerated human hepatocytes is determined as described in McGinnity DF et al. (2004), Evaluation of fresh and cryopreserved hepatocytes as invitro drug metabolism tools for the prediction of metabolic clearance, Drug Metab Dispos, 32:1247-1253. Typically, the assay incubator contains 0.3 μM of the test compound and 10... 6 Cells / mL hepatocytes, with or without pre-incubation with ABT (1 mM) for 0.5 h. Parent compound loss was measured by LC / MS after incubation at 15, 30, 60, and 90 min. Intrinsic clearance rate (μL / min / 10⁻¹⁰) 6 (per cell) is calculated using the following equation for ±ABT. The metabolic fraction of CYP is the percentage of ABT inhibition.

[0336] CL int =-k dep ×Incubation volume / 10 6 cell,

[0337] Where k dep Substrate depletion constant (min) -1 ), is the remaining % of the logarithmic transformation from the y-axis to the time (min) on the x-axis using linear regression. -1 The slope is determined.

[0338] Using recombinant CYP phenotypic analysis (fm) CYP3A4_RCP ) Determine the fraction of CYP3A4-mediated metabolism

[0339] The relative fraction of compound metabolism contributed by a given CYP (e.g., CYP3A4) in the total P450 metabolism of a compound can be determined as follows:

[0340] The metabolism of a group of nine human recombinant CYPs (rCYPs) provided as Supersomes (BD Gentest, Woburn, MA), including rCYP 1A2, 2B6, 2C8, 2C9, 2C19, 2D6, 2J2, 3A4, and 3A5, was evaluated (see Cannady EA et al. (2015) Evacetrapib: in vitro and clinical disposition, metabolism, excretion, and assessment of drug interaction potential with strong CYP3A and CYP2C8 inhibitors, Pharmacol Res Perspect, 3:e00179). Incubation and calculations were performed essentially as described in Wickremsinhe ER et al. (2014), Disposition and metabolism of LY2603618, a Chk-1 inhibitor following intravenous administration in patients with advanced and / or metastatic solid tumors, Xenobiotica 44:827-841, except that incubation was performed for 2 hours to give a low turnover rate of the test compound. The intrinsic clearance rate (CL) of rCYPs was calculated according to the following equation. int ) scaled to human liver microsomes (HLM-scaled CL) int ):

[0341] HLM-Scaled CL int =-k dep ×(incubation volume / pmol rCYP)×RAF

[0342] Where k dep It is the substrate consumption rate constant (min) -1 pmol rCYP is the batch-specific amount of rCYP during incubation, and the relative activity factor (RAF) is the relative activity factor (pmol / mg) suitable for the rCYP / HLM pair. The rate constant was determined experimentally, and the remaining percentage (in min) was log-transformed from the y-axis to the time (in min) on the x-axis using linear regression. -1 Determine the slope. pmol rCYP and RAF are constants provided by the supplier for the supersome.

[0343] Fraction of CYP3A4-mediated metabolism (fm) CYP3A4_RCP) By using the HLM scale of CYP3A4 CL int Divide by the CL on the HLM scale for each P450 subtype as determined. int The sum of these factors determines the outcome (Cannady et al., 2015).

[0344] Hepatocyte metabolite profile

[0345] Hepatocyte metabolomics were determined to confirm metabolite formation via P450-mediated oxidation and phase II enzyme-mediated non-oxidative pathways (e.g., UDP-glucuronyltransferase and sulfotransferase). Its determination in human hepatocytes was essentially as described in Zhou X et al. (2016), *Difference in the Pharmacokinetics and Hepatic Metabolism of Antidiabetic Drugs in Zucker Diabetic Fatty and Sprague-Dawley Rats*, *DrugMetab Disposal* 44:1184-1192. Briefly, incubation was performed at 37°C in a CO2 incubator using 24-well plates containing 250,000 cells / well. A stock solution of the test compound was added to the culture medium to obtain a final incubation concentration of 2 μM. After 4 hours, the incubation, including the culture medium and cells, was quenched with an equal volume of acetonitrile. Samples were processed and analyzed using LC / MS, essentially as described in Zhou et al., 2016.

[0346] Calculate the fraction of compounds cleared by CYP3A4 metabolism (fm) CYP3A4 )

[0347] Basically, based on Burkey JL et al. (2002), Disposition of LY333531, a selective protein kinase C beta inhibitor, in the Fischer 344rat and beagle dog, Xenobiotica 32:1045-1052, the renal and bile excretion of the test compound was tested in cannulated dogs. Basically, as described in test example 7, the renal and bile excretion of the compound was found to be negligible. The fraction of the compound cleared by CYP3A4 metabolism was calculated using the following equation:

[0348] fm CYP3A4 =fm CYP x fm 3A4_RCP

[0349] Where fm CYP In hepatocytes CL intThe fraction of compounds metabolized by CYP measured in ±ABT assays.

[0350] fm 3A4_RCP The fraction of compounds metabolized via CYP3A4 in recombinant CYP phenotypic analysis.

[0351] Determine the fraction of first-pass metabolism evaded (F) G )

[0352] The fraction of metabolism in the gut can be estimated using the following equation found in Yang J et al. (2007), Prediction of intestinal first-pass drug metabolism, Curr Drug Metab 8:676-684:

[0353] F G =Q gut / (Q gut +fu gut x Cl int CYP3A,gut )

[0354] Q gut =Q villi x Cl perm / (Q villi +Cl perm )

[0355] Cl perm =P app ×Intestinal surface area

[0356] Cl int,CYP3A,gut =CL int,CYP3A,liver / fu mic × Microsomal protein in the intestine (mg) × 0.4 (internal calculation)

[0357] Q gut It is a mixed flow term that depends on villous blood flow and the permeability of compounds.

[0358] fu gut It is the fraction of unbound drugs in intestinal cells.

[0359] Q villi It is villous blood flow

[0360] Cl perm It is a term that defines the clearance through the permeability of intestinal cells.

[0361] P app Passive permeability measured in the internal MDCKII cell line

[0362] fu mic It is the fraction of unbound drug in human liver microsomes.

[0363] Human gut parameters were obtained from Yang et al., 2007 and Gertz M et al. (2010), Prediction of human intestinal first-pass metabolism of 25CYP3A substrates from in vitro clearance and permeability data, Drug Metab Dispos 38:1147-1158.

[0364] Mechanostatic model used to evaluate victim DDI suppressed by CYP3A4

[0365] Potential variations in drug AUC exposure of the test compound in the presence of itraconazole can be determined using a mechanostatic model essentially as described by Han B et al. (2013) Prediction of CYP3A Mediated Drug-Drug Interactions: Estimation of Gut Wall and Hepatic Contributions, ASCPT Annual Meeting, Indianapolis, IN.

[0366] AUC PO,inh / AUC PO =[1 / (Ax fm+(1-fm))]x[1 / (X x(1-F G )+F G )]

[0367] A = 1 / (1 + [I]) h / Ki)

[0368] X = 1 / (1 + [I]) gut / Ki)

[0369] Where fm is the fraction of liver CYP3A4 metabolism.

[0370] F G It is the fraction of intestinal cells that escape intestinal metabolism.

[0371] Ki is the dissociation constant of itraconazole from the enzyme.

[0372] [I] h Itraconazole concentration in the liver

[0373] [I] gut Itraconazole concentration in the intestine

[0374] The parameters for itraconazole as a CYP3A4 inhibitor (A = 0.10 and X = 0.10) are from Olkkola KT et al. (1994). Midazolam should be avoided in patients receiving the systemic antimycotics ketoconazole or itraconazole, Clin Pharmacol Ther 55:481-485.

[0375] Example 7, representing the intervention compound, was tested essentially as described above and found to be primarily metabolized via non-oxidative processes in hepatocytes, rather than via oxidative metabolism through P450 (the fraction metabolized by CYP was 32.3%). This compound was not examined in the RCP assay, but a worst-case parameter assuming 100% CYP3A4 metabolism was used to predict the AUC ratio in the presence of itraconazole. The predicted AUC ratio was less than 2. Therefore, the compounds of the present invention are considered to have minimal risk of harmful drug-drug interactions via any CYP, including CYP3A4 (see Table 4).

[0376] Table 4. Example 7 compound fm CYP3A4 F G Compared to the predicted AUC

[0377] .

Claims

1. A compound having the following formula: in: R 1 yes , or ; R 2 It is -F or –Cl; R 3 It is -F or –Cl; and R 4 It is -H or -F; The condition is when R 1 for When, then R 4 It is -F.

2. The compound according to claim 1, having the following formula: in: R 1 yes , , , , or ; R 2 It is -F or –Cl; R 3 It is -F or –Cl; and R 4 It is -H or -F; The condition is when R 1 for When, then R 4 It is -F.

3. The compound according to claim 1, wherein the compound is: 。 4. A pharmaceutical composition comprising a compound according to any one of claims 1 to 3, and a pharmaceutically acceptable excipient.

5. The pharmaceutical composition according to claim 4, comprising: ， And pharmaceutically acceptable excipients.

6. Use of the compound according to any one of claims 1 to 3 in the preparation of a medicament for treating Parkinson's disease or Alzheimer's disease.

7. Use of an effective amount of compound 2-(2-chloro-6-fluoro-phenyl)-1-[(1S,3R)-6-fluoro-3-(hydroxymethyl)-5-(3-hydroxy-3-methyl-butyl)-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl] ethyl ketone in the preparation of a medicament for the treatment of Parkinson's disease in patients in need.

8. Use of an effective amount of compound 2-(2-chloro-6-fluoro-phenyl)-1-[(1S,3R)-6-fluoro-3-(hydroxymethyl)-5-(3-hydroxy-3-methyl-butyl)-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl] ethyl ketone in the preparation of a medicament for the treatment of Alzheimer's disease in patients in need.

9. Use of an effective amount of compound 2-(2-chloro-6-fluoro-phenyl)-1-[(1S,3R)-6-fluoro-3-(hydroxymethyl)-5-(3-hydroxy-3-methyl-butyl)-1-methyl-3,4-dihydro-1H-isoquinoline-2-yl] ethyl ketone in the preparation of a medicament for the treatment of dopaminergic CNS disorders in patients in need.

10. Use of the compound according to any one of claims 1 to 3 and the dopamine precursor in the preparation of a medicament for treating patients with dopaminergic CNS disorders simultaneously, separately, or in sequence.

11. Use of the compound according to any one of claims 1 to 3 and the dopamine agonist in the preparation of a medicament for treating patients with dopaminergic CNS disorders simultaneously, separately, or in sequence.

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