COMPOUND AND PHARMACEUTICAL COMPOSITION OF INDOLINE DERIVATIVES REPLACED AS INHIBITORS OF DENGUE VIR

专利摘要:
The present invention relates to substituted indoline derivatives, methods to prevent or treat viral dengue infections when using said compounds and also relates to said compounds for use as a medicine, more preferably for use as a medicine to treat or prevent dengue viral infections. The present invention also relates to pharmaceutical compositions or preparations for combining the compounds, with the compositions or preparations for use as a medicament, more preferably for the prevention or treatment of dengue viral infections. The invention also relates to processes for preparing the compounds.
公开号:BR112019024195A2
申请号:R112019024195-7
申请日:2018-05-18
公开日:2020-06-23
发明作者:Bonfanti Jean-François；Jean-François Bonfanti；Rudolf Romanie Kesteleyn Bart；Bart Rudolf Romanie Kesteleyn；Alice Marie-Eve Bardiot Dorothée；Dorothée Alice Marie-Eve Bardiot；Didier M. Marchand Arnaud；Arnaud Didier M. Marchand；Coesemans Erwin；Erwin Coesemans；Michel Claude Fortin Jérôme；Jérôme Michel Claude Fortin；Jean Maurice MERCEY Guillaume；Guillaume Jean Maurice Mercey；Jean-Marie Bernard Raboisson Pierre；Pierre Jean-Marie Bernard Raboisson
申请人:Janssen Pharmaceuticals, Inc.；Katholieke Universiteit Leuven；
IPC主号:

专利说明:

[001] [001] The present invention relates to substituted indoline derivatives, with methods to prevent or treat dengue viral infections when using said compounds and also refers to said compounds for use as a medicine, more preferably for use as a medicine to treat or prevent dengue viral infections. The present invention also relates to pharmaceutical compositions or preparations for combining the compounds, with the compositions or preparations for use as a medicament, more preferably for the prevention or treatment of dengue viral infections. The invention also relates to processes for preparing the compounds. BACKGROUND OF THE INVENTION
[002] [002] Flaviviruses, which are transmitted by mosquitoes or ticks, cause life-threatening infections in humans, such as encephalitis and hemorrhagic fever. Four distinct but closely related serotypes of dengue flavivirus are known, so-called DENV-1, -2, -3 and -4. Dengue is endemic in most tropical and subtropical regions around the world, predominantly in urban and semi-urban areas. According to the World Health Organization (WHO), 2.5 billion people, of which 1 billion children are at risk of DENV infection (WHO, 2002). 50 to 100 million estimated cases of dengue fever [DF], half a million cases of severe dengue disease (ie, dengue hemorrhagic fever [DHF] and dengue shock syndrome [DSS]), and more than 20,000 deaths occur globally each year. DHF has become a leading cause of hospitalization and death among children in endemic regions. Altogether, dengue represents the most common cause of arboviral disease. Due to large recent outbreaks in countries located in Latin America, Southeast Asia and the Western Pacific (including Brazil, Puerto Rico, Venezuela, Cambodia, Indonesia, Vietnam, Thailand), the numbers of dengue cases have increased dramatically over the past few years . Not only is the number of dengue cases increasing as the disease is spreading to new areas, but outbreaks also tend to be more serious.
[003] [003] After infection with another serotype, the pre-existing heterologous antibodies form complexes with the newly infected dengue virus serotype but do not neutralize the pathogen. Instead, it is believed that virus entry into cells is facilitated, resulting in uncontrolled virus replication and higher peak viral titers. In both primary and secondary infections, higher viral titers are associated with more severe dengue disease. Since maternal antibodies can easily pass to breastfeeding children, this may be one of the reasons why children are more affected by severe dengue disease than adults.
[004] [004] In locations with two or more serotypes circulating simultaneously, also referred to as hyper-endemic regions, the risk of severe dengue disease is significantly higher due to an increased risk of experiencing a secondary, more serious infection. Furthermore, in a situation of hyperendemicity, the likelihood of the emergence of more virulent strains is increased, which in turn increases the likelihood of dengue hemorrhagic fever (DHF) or dengue shock syndrome.
[005] [005] Mosquitoes that carry dengue, including Aedes aegypti and Aedes albopictus (tiger mosquito), are moving north across the globe. According to the Centers for Disease Control and
[006] [006] Dengvaxia®, the dengue vaccine produced by Sanofi Pasteur, was first approved in Mexico and has since been approved in more countries. Nevertheless, the vaccine leaves plenty of room for improvement due to its limited efficacy, especially against DENV-1 and -2, low efficacy in patients without previous treatment of flavivirus and the prolonged dosing schedule.
[007] [007] Despite these shortcomings, the vaccine is a major advance in endemic definitions, since it will offer protection to a large part of the population, but probably not to very young children, on whom the greatest incidence of dengue falls. In addition, the dosing schedule and the very limited efficacy in subjects without previous treatment of flavivirus make it inadequate and probably not compensatory / profitable for travelers from non-endemic areas to dengue-endemic areas. The aforementioned shortcomings of dengue vaccines are the reason why there is a need for a pre-exposure prophylactic dengue antiviral.
[008] [008] Furthermore, today, specific antiviral drugs are not available for the treatment or prevention of infection by the dengue fever virus. There is clearly still a great unmet medical need for therapies for the prevention or treatment of viral infections in animals, more particularly in humans and especially for viral infections caused by flaviviruses, more particularly dengue viruses. Compounds with good antiviral potency, no or low levels of side effects, broad spectrum activity against multiple dengue virus serotypes, low toxicity and / or good pharmacokinetic or dynamic properties are highly needed.
[009] [009] WO-2010/021878 describes 2-phenylpyrrolidine and indoline derivatives as cold menthol receptor antagonists for the treatment of inflammatory and central diseases. WO-2013/045516 describes indole and indoline derivatives for use in the treatment of dengue viral infections.
[0010] [0010] The present invention now provides compounds, substituted indoline derivatives, which show high potent activity against all four (4) dengue virus serotypes. SUMMARY OF THE INVENTION
[0011] [0011] The present invention is based on the unexpected discovery that at least one of the problems mentioned above can be solved by the current compounds of the invention.
[0012] [0012] The present invention provides compounds that have been shown to have potent antiviral activity against all four (4) currently known serotypes. The present invention further demonstrates that these compounds efficiently inhibit the proliferation of the dengue virus (DENV). Therefore, these compounds constitute a useful class of potent compounds that can be used in the treatment and / or prevention of viral infections in animals, mammals and humans, more specifically for the treatment and / or prevention of infections with dengue virus.
[0013] [0013] The present invention also relates to the use of such compounds as medicines and their use for the manufacture of medicines for the treatment and / or prevention of viral infections, in particular with viruses belonging to the family of dengue viruses in animals or mammals, more particularly in humans. The invention also relates to methods for the preparation of all such compounds and to pharmaceutical compositions comprising them in an effective amount.
[0014] [0014] The present invention also relates to a method of treating or preventing viral infections of dengue in humans by administering an effective amount of one or more such compounds, or a pharmaceutically acceptable salt thereof, optionally in combination with a or more other drugs, such as another antiviral agent, to a patient in need.
[0015] [0015] One aspect of the invention is the supply of compounds of formula (I), including any stereochemically isomeric form thereof: Cl CH3
[0016] [0016] The compounds specifically mentioned above are selected from the group comprising: Cl Cl OMe OMe O O F O N F O N N H N H CUTE F OH F OH
[0017] [0017] A first group of compounds are compounds of the formula (I) in which A represents - (CH2) n- in which n is 3 or 4.
[0018] [0018] A second group of compounds are compounds of the formula (I) in which A represents -O- (CH2) n- in which n is 2 or 4.
[0019] [0019] A third group of compounds are compounds of formula (I) in which A represents
[0020] [0020] -O- (CH2) n- where n is 3 and one or two CH2 are replaced by one or two CH3; or A represents.
[0021] [0021] A fourth group of compounds are compounds of the formula (I) in which A represents -CH2-O- (CH2) n- in which n is 2.
[0022] [0022] A fifth group of compounds are compounds of the formula (I) where A represents -X-Y- where X is -O-, -OCH2- or -NH-; and Y is C3-4 cycloalkyl optionally substituted by fluorine.
[0023] [0023] A sixth group of compounds are compounds of the formula (I) where A represents -X-Y- where X is -O-, -OCH2- or -NH-; and Y is a bicyclist [1.1.1] pentanyl.
[0024] [0024] A seventh group of compounds are compounds of the formula (I) in which R1 is trifluoromethoxy, R2 is hydrogen and R3 is hydrogen.
[0025] [0025] An eighth group of compounds are compounds of the formula (I) in which R1 is trifluoromethoxy, R2 is hydrogen and R3 is methoxy.
[0026] [0026] A ninth group of compounds are compounds of the formula (I) in which R1 is trifluoromethyl, R2 is methoxy and R3 is hydrogen.
[0027] [0027] A tenth group of compounds are compounds of formula (I) in which R1 is trifluoromethoxy, R2 is fluorine and R3 is hydrogen.
[0028] [0028] An eleventh group of compounds are compounds of the formula (I) in which R1 is chlorine, R2 is methoxy and R3 is hydrogen.
[0029] [0029] Part of the current invention is also a pharmaceutical composition comprising a compound mentioned above or a stereoisomeric form thereof, salt, solvate or pharmaceutically acceptable polymorph together with one or more pharmaceutically acceptable excipients, diluents or carriers.
[0030] [0030] The pharmaceutically acceptable salts of said compounds include their acid and base addition salts. Suitable acid addition salts are formed from acids that form non-toxic salts. Suitable base salts are formed from bases that form non-toxic salts.
[0031] [0031] The pharmaceutically acceptable acid salts as mentioned above are intended to comprise the therapeutically active non-toxic acid addition salt forms that the compounds of formula (I) are capable of forming. These pharmaceutically acceptable acid addition salts can be conveniently obtained by treating the base form with such an appropriate acid. Suitable acids comprise, for example, inorganic acids such as hydrochloric acids, e.g., hydrochloric or hydrobromic acid, sulfuric, nitric, phosphoric acid and the like; or organic acids such as, for example, acetic, propanoic, hydroxyacetic, lactic, pyruvic, oxalic (ie, ethanedioic), malonic, succinic (ie, butanedioic acid), maleic, fumaric, malic, tartaric, citric, methanesulfonic, ethanesulfonic , benzenesulfonic, p-toluenesulfonic, cyclamic, salicylic, p-aminosalicylic, pamoic and similar acids.
[0032] [0032] The compounds of the invention can also exist in unsolvated and solvated forms. The term "solvate" is used herein to describe a molecular complex comprising the compound of the invention and one or more pharmaceutically acceptable solvent molecules, for example, ethanol.
[0033] [0033] The term "polymorph" refers to the ability of the compound of the invention to exist in more than one crystal shape or structure.
[0034] [0034] The compounds of the present invention can be administered as crystalline or amorphous products. They can be obtained, for example, as solid buffers, powders or films by methods such as precipitation, crystallization, lyophilization, spray drying or evaporative drying. They can be administered alone or in combination with one or more other compounds of the invention or in combination with one or more other drugs. They will generally be administered as a formulation in combination with one or more pharmaceutically acceptable excipients. The term "excipient" is used throughout this document to describe any ingredient other than the compound (s) of the invention. The choice of excipient depends largely on factors such as the particular mode of administration, the effect of the excipient on solubility and stability and the nature of the dosage form.
[0035] [0035] The compounds of the present invention or any subgroup thereof can be formulated in various dosage forms for administration purposes. As appropriate compositions, all compositions usually used for systemic drug administration can be cited. To prepare the pharmaceutical compositions of this invention, an effective amount of the particular compound, optionally in the form of addition salt, as the active ingredient is combined in admixture with a pharmaceutically acceptable carrier, which carrier can take a wide variety of forms depending on the form of preparation desired for administration. These pharmaceutical compositions are desirably in unit dosage form suitable, for example, for oral or rectal administration. For example, in the preparation of compositions in oral dosage form, any of the usual pharmaceutical means such as, for example, water, glycols, oils, alcohols and the like can be used in the case of liquid oral preparations such as suspensions, syrups, elixirs , emulsions, and solutions; or solid carriers such as starches, sugars, kaolin, thinners, lubricants, binders, disintegrating agents and the like in the case of powders, pills, capsules and tablets. Due to their ease of administration, tablets and capsules represent the most advantageous oral dosage unit forms, in which case, of course, solid pharmaceutical carriers are used. Also included are preparations in solid form that can be converted, immediately before use, to liquid forms.
[0036] [0036] It is especially advantageous to formulate the pharmaceutical compositions mentioned above in unit dosage form for ease of administration and uniformity of dosage. Unit dosage form as used herein refers to physically discrete units suitable as unitary dosages, each unit containing a predetermined amount of active ingredient calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. Examples of such unit dosage forms are tablets (including grooved or coated tablets), capsules, pills, powder packs, cachets, suppositories, injectable solutions and suspensions and the like and their multiple secretions.
[0037] [0037] Experts in the treatment of infectious diseases will be able to determine the effective amount from the test results presented hereinafter. In general it is contemplated that an effective daily amount would be 0.01 mg / kg to 50 mg / kg of body weight, more preferably 0.1 mg / kg to 10 mg / kg of body weight. It may be appropriate to administer the required dose as two, three, four or more underdoses at appropriate intervals throughout the day. Said underdoses can be formulated as unit dosage forms, for example, containing 1 to 1000 mg and, in particular, 5 to 200 mg of active ingredient per unit dosage form.
[0038] [0038] The exact dosage and frequency of administration depend on the particular compound of the invention used, the particular condition being treated, the severity of the condition being treated, the age, weight and general physical condition of the particular patient as well as other medication that the individual may be taking, as is well known to those skilled in the art. Furthermore, it is evident that the effective amount can be decreased or increased depending on the response of the treated subject and / or depending on the physician's judgment prescribing the compounds of the present invention. The ranges of effective amounts mentioned above are therefore guidelines only and are not intended to limit the scope or use of the invention to any extent.
[0039] [0039] The present disclosure is also intended to include any isotopes of atoms present in the compounds of the invention. For example, hydrogen isotopes include tritium and deuterium and carbon isotopes include C-13 and C-14.
[0040] [0040] As used herein, any chemical formula with bonds shown only as solid lines and not as solid minted or perforated minted bonds, or otherwise indicated as having a particular configuration (eg, R, S) in around one or more atoms, contemplate each possible stereoisomer, or mixture of two or more stereoisomers.
[0041] [0041] Formerly and henceforth, the terms "compound of the formula
[0042] [0042] The terms "stereoisomers", "stereoisomeric forms" or "stereochemically isomeric forms" previously or hereinafter are used interchangeably.
[0043] [0043] The invention includes all stereoisomers of the compounds of the invention as pure stereoisomers or as a mixture of two or more stereoisomers. Enantiomers are stereoisomers that are non-overlapping images in each other's mirror. A 1: 1 mixture of a pair of enantiomers is a racemate or racemic mixture. Diastereomers (or diastereoisomers) are stereoisomers that are not enantiomers, i.e., they are not related as mirror images. Substituents in bivalent cyclic (partially) saturated radicals can be in the cis or trans configuration; for example, if a compound contains a disubstituted cycloalkyl group, the substituents can be in the cis or trans configuration.
[0044] [0044] The term "stereoisomers" also includes any rotamers, also called conformational isomers, that the compounds of formula (I) can form.
[0045] Therefore, the invention includes enantiomers, diastereomers, racemates, E isomers, Z isomers, cis isomers, trans isomers, rotamers and any mixture thereof, whenever chemically possible.
[0046] [0046] The meaning of all these terms, i.e., enantiomers, diastereomers, racemates, cis isomers, trans isomers and mixtures thereof are known to the person skilled in the art.
[0047] [0047] The absolute configuration is specified according to the Cahn-Ingold-Prelog system. The configuration on an asymmetric atom is specified by R or S. The resolved stereoisomers whose absolute configuration is not known can be called
[0048] [0048] When a specific stereoisomer is identified, this means that said stereoisomer is substantially free, ie, associated with less than 50%, preferably less than 20%, more preferably less than 10%, even more preferably less than than 5%, in particular less than 2% and most preferably less than 1%, of the other stereoisomers. Thus, when a compound of formula (I) is for example specified as (R), this means that the compound is substantially free of the (S) isomer; when a compound of formula (I) is for example specified as E, this means that the compound is substantially free of the Z isomer; when a compound of formula (I) is for example specified as cis, this means that the compound is substantially free of the trans isomer.
[0049] [0049] Some of the compounds according to formula (I) can also exist in their tautomeric form. Such forms, to the extent that they may exist, although not explicitly indicated in formula (I) above, are intended to be included in the scope of the present invention.
[0050] [0050] The compounds of formula (I) of the present invention all have at least one asymmetric carbon atom as indicated in the figure below by the carbon atom marked with *: Cl CH3
[0051] [0051] Due to the presence of said chiral center, a "compound of the formula (I)" can be the enantiomer (R), the enantiomer (S), the racemic form or any possible combination of the two individual enantiomers for any reason. When the absolute configuration of an asymmetric carbon atom was not known, a relative stereochemistry descriptor was used: * R or * S (or R * and S *) to indicate the pure but unknown stereochemistry of the chiral center.
[0052] [0052] Since radical A allows substitutions by introducing asymmetric carbon atoms, the compounds of formula (I) can have more than one asymmetric carbon atom. When the absolute stereochemistry of more than one asymmetric carbon atoms has not been determined, relative stereochemistry has been indicated using the relative stereochemistry descriptors * R and * S and where possible in combination with cis and trans when radical A contains a cyclic moiety .
[0053] [0053] In one aspect, the present invention relates to a first group of compounds of formula (I) in which the compounds of formula (I) have specific rotation (-).
[0054] [0054] In a further aspect, the present invention relates to a second group of compounds of the formula (I) in which the compounds of the formula (I) have the specific rotation (+).
[0055] [0055] In one embodiment, the present invention relates to a compound of formula (I) having the specific rotation (+) in which said compound is selected from the group consisting of compounds (1C), (1D), (2A ), (4C), (4D), (5A), (6AB), (6BB), (7B), (8B), (9B), (10B), (11B), (12B), (13B), (14A), (15B), (17C), (17D), (18B), (19AB), (19BB), (20C), (20D), (21B), (22AB), (22BB), (23B ), (24B), (25B), (27B), (28AB), (28BB), (29AB), (29BB), (30A), (31A), (32B), (33C), and (33D) . Examples LC / MS Methods
[0056] [0056] Measurement by High Performance Liquid Chromatography
[0057] [0057] The flow from the column was conducted to the Mass Spectrometer (MS) that was configured with an ion source at atmospheric pressure. It is within the knowledge of the person skilled in the art to define the adjustment parameters (eg, scanning range, dwell time ...) in order to obtain ions allowing the identification of the nominal monoisotopic molecular weight (PM) of the compound. Data acquisition was performed with appropriate software.
[0058] [0058] The compounds are described by their retention times (Tr) and ions. If not specified differently in the data table, the reported molecular ion corresponds to [M + H] + (protonated molecule) and / or [M-H] - (deprotonated molecule). In case the compound was not directly ionizable, the type of adduct is specified (i.e., [M + NH4] +, [M + HCOO] -, etc ...). In the case of molecules with multiple isotopic patterns (Br, Cl), the reported value is that obtained for the lowest isotopic mass. All results were obtained with experimental uncertainties that are commonly associated with the method used.
[0059] [0059] Hereinafter, "SQD" stands for Single Quadropole Detector, "MSD" Selective Mass Detector, "TA" at room temperature, "Ethyl siloxane / silica bridge hybrid", "DAD" Diodes Array Detector, "HSS "High Resistance silica.
[0060] [0060] LC / MS method codes (Flow expressed in mL / min; column temperature (T) in ° C; Operating time in minutes).
[0061] [0061] The measurement by SFC was performed using an Analytical chromatography system with Supercritical fluid (SFC) composed of a binary pump for administration of carbon dioxide (CO2) and modifier, a self-sampler, a column oven, an array detector diode equipped with a high pressure flow cell resisting up to 400 bar. If configured with a Mass Spectrometer (MS), the flow from the column was conducted to the (MS). It is within the knowledge of the person skilled in the art to define the adjustment parameters (eg, scanning range, dwell time ...) in order to obtain ions allowing the identification of the nominal monoisotopic molecular weight (PM) of the compound. Data acquisition was performed with appropriate software.
[0062] [0062] Analytical SFC / MS Methods (Flow expressed in mL / min; column temperature (T) in ° C; Operating time in minutes, Backpressure (BPR) in bars). Flow time Code of operation column mobile phase gradient --------- method ------------ T of Col
[0063] [0063] The values are peak values or melting ranges and are obtained with experimental uncertainties that are commonly associated with this analytical method. DSC823e (indicated as DSC)
[0064] [0064] For a composite number, the melting points were determined with a DSC823e (Mettler-Toledo). Melting points were measured with a temperature gradient of 10 ° C / minute. The maximum temperature was 300 ° C. Optical Rotations:
[0065] [0065] Optical rotations were measured on a PerkinElmer 341 polarimeter with a sodium lamp and reported as follows: [Į] º (Ȝ, c g / 100 mL, solvent, T ° C).
[0066] [0066] [Į] ȜT = (100Į) / (lxc): where l is the path length in dm and c is the concentration in g / 100 mL for a sample at a temperature T (° C) and a wavelength Ȝ (in nm). If the wavelength of light used is 589 nm (line D of sodium), then the symbol D could be used instead. The rotation sign (+ or -) must always be given. When using this equation, the concentration and solvent are always provided in parentheses after rotation. Rotation is reported using degrees and no concentration units are given (assumed to be g / 100 mL).
[0067] [0067] Stereochemistry note: In the examples below, the stereochemistry indications * R and * S refer to pure stereochemistry but unknown to chiral centers. Abbreviations used in the experimental part (M + H) + protonated molecular ion iPrNH2 isopropylamine MH + aq. aqueous iPrOH 2-propanol Boc tert-butyloxycarbonyl K2CO3 potassium carbonate Boc2O di-tert-butyl dicarbonate KNO3 potassium nitrate l broad LiAlH4 aluminum hydride and lithium acetonitrile m / z mass to CH3CN charge CHCl3 chloroform Me methyl CH2Cl2 methyl chloride methanol CH3OH methanol MgSO4 magnesium sulfate CO2 carbon dioxide min minute (s) CsCO3 cesium carbonate MTBE methyl tert-butyl ether d doublet N2 nitrogen DCM dichloromethane Na2CO3 sodium carbonate DIEA diisopropylethylamine Na2SO4 sodium sulfate DIPE diisopropyl ether NaBH4 sodium borohydride DMA dimethylacetamide NaCl sodium chloride DMAP 4-dimethylaminopyridine NaHCO3 sodium bicarbonate DME 1,2-dimethoxyethane NaOH sodium hydroxide DMF dimethylformamide NH4Cl ammonium chloride DMSO dimethyl sulfoxide NH4HCO3 3-ammonium bicarbonate (3-dimethylamino-EDCl NMP N-methylpyrrolidone propyl) carbodiimide eq. equivalent q quartet diethyl ether rt or RT room temperature Et2O triethylamine 2- Et3N chloride SEMCl (trimethylsilyl) ethoxymethyl EtOAc ethyl acetate s singlet EtOH ethanol t hydrogen triple tBuOK potassium H2 tert-butanolate HNO3 nitric acid TEA triethylamine H2O water TEA triethylamine H2O trifluoroacetic H2SO4 sulfuric acid THF tetrahydrofuran hexafluorophosphate O- (7- aza-1H-benzotriazol-1-yl) - HATU 2-Me-THF 2-methyltetrahydrofuran N, N, N ', N'-tetramethyl-uronium - CAS [148893-10-1] HCl hydrochloric acid TMSCl trimethylsilyl chloride HPLC liquid chromatography TMSCF3 trifluoromethyltrimethylsilane high performance Example 1: 4- (3 - ((1- (4-chlorophenyl) -2-oxo-2 acid synthesis - (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) -2-methylbutanoic (Compound 1) and separation in Stereoisomers 1A, 1B, 1C and 1D Synthesis of intermediate 1a:
[0068] [0068] To a stirred solution of tert-butyl 4-bromo-2-methylbutanoate [CAS 1210410-44-8] (1.0 g, 4.22 mmol) in DMF (15 mL) was added 3-amino- 5-methoxyphenol [CAS 162155-27-3] (587 mg, 4.22 mmol) and Cs2CO3 (2.75 g, 8.43 mmol). The reaction was stirred at 60 ° C for 65 h, and allowed to reach room temperature. The mixture was poured into H2O (100 ml). The product was extracted with CH2Cl 2 (2 times). The combined organic layers were dried over MgSO4, filtered, and the solvent was evaporated under reduced pressure. The residue was purified by flash chromatography on silica gel (50 g) using a 100/0 to 50/50 heptane / EtOAc gradient. The desired fractions were combined, evaporated under reduced pressure and coevaporated with CH3CN, yielding tert-butyl 4- (3-amino-5-methoxyphenoxy) -2-methylbutanoate 1a (440 mg). Synthesis of intermediate 1b:
[0069] [0069] A mixture of 6- (trifluoromethoxy) indoline [CAS 959235-95-1] (5 g, 24.6 mmol), 2- (4-chlorophenyl) acetic acid [CAS 1878-66-6] (4, 2 g, 24.6 mmol), HATU (14.3 g, 36.9 mmol) and diisopropylethylamine (12.2 mL, 73.8 mmol) in DMF (60 mL) was stirred at room temperature for 20 h . The mixture was slowly poured into stirring H2O (275 ml) and the resulting suspension was stirred for 50 minutes. The solids were filtered off and washed (4x) with H2O. The solid residue was taken up in toluene (125 ml), filtered over a paper filter, and the filtrate was evaporated under reduced pressure. The solid residue was stirred in Et2O / heptane 2/1 (30 mL), filtered off, washed (3x) with Et2O / heptane 1/1 and dried under vacuum at 50 ° C to provide 2- (4-chlorophenyl) - 1- (6- (trifluoromethoxy) indolin-1-yl) ethanone 1b (7.33 g). Synthesis of intermediate 1c:
[0070] [0070] At -70 ° C, under a flow of N 2, 1 M LiHMDS in THF (41.2 mL, 41.2 mmol) was added to a solution of 2- (4-chlorophenyl) -1- (6 - (trifluoromethoxy) indolin-1-yl) ethanone 1b (7.33 g, 20.6 mmol) in 2-Me-THF (300 ml). The mixture was stirred for 50 min at -70 ° C and trimethylsilyl chloride (4.21 ml, 33.0 mmol) was added slowly. Stirring was continued at -70 ºC for 35 min and a solution of N-
[0071] [0071] A mixture of 2-bromo-2- (4-chlorophenyl) -1- (6- (trifluoromethoxy) indolin-1-yl) ethanone 1c (850 mg, 1.96 mmol), 4- (3-amino Tert-butyl-5-methoxyphenoxy) -2-methylbutanoate 1a (620 mg, 2.10 mmol) and diisopropylethylamine (506 µL, 2.93 mmol) in CH3CN (30 mL) was stirred at 60 ° C for 18 H. The mixture was allowed to reach room temperature and was poured into water (125 ml). The product was extracted (2x) with Et2O. The combined organic layers were washed with brine, dried over MgSO4, filtered off and evaporated under reduced pressure. The residue was purified by flash chromatography on silica gel (12 g) using a gradient of heptane / EtOAc / EtOH 100/0/0 to 40/45/15. The desired fractions were combined and evaporated under reduced pressure and coevaporated with dioxane to provide 4- (3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl ) tert -butyl amino) -5-methoxyphenoxy) -2-methylbutanoate 1d (1.27 g). Synthesis of Compound 1 and separation in Stereoisomers 1A, 1B, 1C and 1D:
[0072] [0072] A chilled (ice bath) solution of 4- (3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) - 5- tert-butyl methoxyphenoxy) -2-methylbutanoate 1d (1.27 g, 1.96 mmol) in
[0073] [0073] The 4 stereoisomers of Compound 1 (900 mg) were separated by preparative chiral SFC (Stationary phase: Chiralpak® Diacel AD 20 x 250 mm, mobile phase: CO2, iPrOH + iPrNH2 0.4%). The product fractions were combined and evaporated under reduced pressure. The stereoisomers in the product fractions of the first two eluted peaks were not completely separated and required additional separation through preparative chiral SFC (Stationary phase: Chiralpak® Diacel AD 20 x 250 mm, mobile phase: CO2, EtOH + 0.4% iPrNH2 ). The product fractions were combined and evaporated under reduced pressure.
[0074] [0074] The stereoisomer eluted first was purified by flash chromatography on silica gel (12 g) with a gradient of heptane / EtOAc / EtOH / HOAc 100/0/0/0 to 40/45 / 14.7 / 0, 3. The desired fractions were combined and evaporated under reduced pressure and coevaporated with CH3CN. The product was lyophilized from a solvent mixture of CH3CN (2 ml) and H2O (1.2 ml) to provide Stereoisomer 1A (63 mg).
[0075] [0075] The stereoisomer eluted in second place was purified by flash chromatography on silica gel (12 g) with a gradient of heptane / EtOAc / EtOH / HOAc 100/0/0/0 to 40/45 / 14.7 / 0, 3. The desired fractions were combined and evaporated and coevaporated with CH3CN. The product was lyophilized from a solvent mixture of CH3CN (2 ml) and H2O (1.2 ml) to provide Stereoisomer 1B (79 mg).
[0076] [0076] The stereoisomer eluted in third place was purified by preparative HPLC (Stationary phase: RP XBridge® Prep C18 OBD - 10 µm, 30 x 150 mm, mobile phase: 0.25% NH4HCO3 solution in water, CH3CN). The desired fractions were combined and the volatile organics were evaporated under reduced pressure. The residue was mixed with EtOAc (25 ml) and 1 N HCl (0.5 ml). After stirring for 10 min, the layers were separated. The organic layer was isolated, washed with brine, dried over MgSO4, filtered, evaporated under reduced pressure and coevaporated with CH3CN. The residue was lyophilized from a solvent mixture of CH3CN (1.5 ml) and H2O (0.75 ml) to provide Stereoisomer 1C (62 mg).
[0077] [0077] The stereoisomer eluted in fourth place was purified by flash chromatography on silica gel (12 g) with a gradient of heptane / EtOAc / EtOH / HOAc 100/0/0/0 to 40/45 / 14.7 / 0, 3. The desired fractions were combined and evaporated under reduced pressure and coevaporated with CH3CN. The product was lyophilized from a solvent mixture of CH3CN (2 mL) and H2O (1.2 mL) to provide Stereoisomer 1D (105 mg) Compound 1: 1
[0078] [0078] H NMR (400 MHz, DMSO-d6) į ppm 1.10 (dd, J = 7.0, 1.3 Hz, 3 H) 1.69 (dq, J = 13.6, 6.7 Hz, 1 H) 1.91 - 2.01 (m, 1 H) 2.43 - 2.48 (m, 1 H) 3.07 - 3.26 (m, 2 H) 3.61 (s, 3 H) 3.85 (tl, J = 6.5 Hz, 2 H) 4.04 (td, J = 10.3, 7.2 Hz, 1 H) 4.52 (td, J = 10.2 , 6.4 Hz, 1 H) 5.56 (d, J = 8.8 Hz, 1 H) 5.75 (t, J = 2.0 Hz, 1 H) 5.89 - 5.98 (m , 2 H) 6.43 (d, J = 8.6 Hz, 1 H) 7.01 (dd, J = 8.3, 1.4 Hz, 1 H) 7.33 (d, J = 8, 4 Hz, 1 H) 7.40 - 7.47 (m, 2 H) 7.51 - 7.58 (m, 2 H) 8.03 (s, 1 H) 12.16 (sl, 1 H)
[0079] [0079] LC / MS (LC-A method): Rt 1.14 min, MH + 593 Stereoisomer 1A: 1
[0080] [0080] H NMR (400 MHz, DMSO-d6) į ppm 1.10 (d, J = 7.0 Hz, 3 H) 1.69 (dq, J = 13.5, 6.6 Hz, 1 H ) 1.91 - 2.01 (m, 1 H) 2.46 - 2.48 (m, 1 H)
[0081] [0081] LC / MS (LC-A method): Rt 1.15 min, MH + 593
[0082] [0082] [Į] D20: -37.6 ° (c 0.415, DMF)
[0083] [0083] Chiral SFC (SFC-A method): Rt 3.52 min, MH + 593 100% chiral purity. Stereoisomer 1B: 1
[0084] [0084] H NMR (400 MHz, DMSO-d6) į ppm 1.09 (d, J = 7.0 Hz, 3 H) 1.69 (dq, J = 13.6, 6.6 Hz, 1 H ) 1.91 - 2.01 (m, 1 H) 2.44 - 2.48 (m, 1 H) 3.08 - 3.27 (m, 2 H) 3.61 (s, 3 H) 3 , 79 - 3.90 (m, 2 H) 4.04 (td, J = 10.4, 7.2 Hz, 1 H) 4.52 (td, J = 10.2, 6.6 Hz, 1 H) 5.55 (d, J = 8.8 Hz, 1 H) 5.75 (t, J = 2.0 Hz, 1 H) 5.92 - 5.97 (m, 2 H) 6.43 (d, J = 8.8 Hz, 1 H) 7.01 (dd, J = 8.1, 1.5 Hz, 1 H) 7.33 (d, J = 8.1 Hz, 1 H) 7 , 44 (d, J = 7.8 Hz, 2 H) 7.55 (d, J = 7.2 Hz, 2 H) 8.03 (s, 1 H) 12.16 (sl, 1 H)
[0085] [0085] LC / MS (LC-A method): Rt 1.15 min, MH + 593
[0086] [0086] [Į] D20: -65.3 ° (c 0.455, DMF)
[0087] [0087] Chiral SFC (SFC-A method): Rt 4.15 min, MH + 593 chiral purity 97.1%. Stereoisomer 1C: 1
[0088] [0088] H NMR (400 MHz, DMSO-d6) į ppm 1.11 (d, J = 7.0 Hz, 3 H) 1.70 (dq, J = 13.5, 6.5 Hz, 1 H ) 1.90 - 2.03 (m, 1 H) 2.44 - 2.49 (m, 1 H) 3.07 - 3.25 (m, 2 H) 3.62 (s, 3 H) 3 , 86 (t, J = 6.6 Hz, 2 H) 3.98 - 4.11 (m, 1 H) 4.46 - 4.57 (m, 1 H) 5.56 (d, J = 8 , 6 Hz, 1 H) 5.76 (t, J = 2.1 Hz, 1 H) 5.90 - 5.99 (m, 2 H) 6.44 (d, J = 8.8 Hz, 1 H) 7.01 (dd, J = 7.9, 1.8 Hz, 1 H) 7.34 (d, J = 8.4 Hz, 1 H) 7.44 (d, J = 8.4 Hz , 2 H) 7.55 (d, J = 7.8 Hz, 2 H) 8.04 (s, 1 H) 12.18 (ls, 1 H)
[0089] [0089] LC / MS (LC-A method): Rt 1.15 min, MH + 593
[0090] [0090] [Į] D20: + 35.2 ° (c 0.455, DMF)
[0091] [0091] Chiral SFC (SFC-A method): Rt 2.84 min, MH + 593 chiral purity 99.3%. Stereoisomer 1D: 1
[0092] [0092] H NMR (400 MHz, DMSO-d6) į ppm 1.10 (d, J = 7.0 Hz, 3 H) 1.70 (dq, J = 13.5, 6.6 Hz, 1 H ) 1.92 - 2.02 (m, 1 H) 2.46 - 2.49 (m, 1 H) 3.09 - 3.29 (m, 2 H) 3.62 (s, 3 H) 3 , 80 - 3.92 (m, 2 H) 4.05 (td, J = 10.5, 7.0 Hz, 1 H) 4.53 (td, J = 10.4, 6.5 Hz, 1 H) 5.56 (d, J = 8.8 Hz, 1 H) 5.76 (t, J = 2.0 Hz, 1 H) 5.93 - 5.97 (m, 2 H) 6.44 (d, J = 8.6 Hz, 1 H) 7.01 (dd, J = 8.1, 1.5 Hz, 1 H) 7.34 (d, J = 8.1 Hz, 1 H) 7 , 40 - 7.47 (m, 2 H) 7.56 (d, J = 8.4 Hz, 2 H) 8.04 (s, 1 H) 12.17 (ls, 1 H)
[0093] [0093] LC / MS (LC-A method): Rt 1.15 min, MH + 593
[0094] [0094] [Į] D20: + 64.3 ° (c 0.42, DMF)
[0095] [0095] Chiral SFC (SFC-A method): Rt 2.65 min, MH + 593 chiral purity 98.1%. Example 2: 4- (3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) -2 acid synthesis , 2-dimethylbutanoic (Compound 2) and chiral separation in Enantiomers 2A and 2B Synthesis of intermediate 2a:
[0096] [0096] To a stirred solution of methyl 4-bromo-2,2-dimethylbutanoate [CAS 4833-99-2] (2.5 g, 12 mmol) in DMF (35 mL) were added 3-amino-5- methoxyphenol [CAS 162155-27-3] (1.66 g, 12 mmol) and Cs2CO3 (7.79 g, 23.9 mmol). The reaction was stirred at 60 ° C for 65 h, and allowed to reach room temperature. The mixture was poured into H2O (150 ml). The product was extracted with CH2Cl2. The organic layer was dried over MgSO4, filtered, and the solvent was evaporated under reduced pressure. The residue was purified by flash chromatography on silica gel (25 g) using a gradient from heptane / CH2Cl2 / MeOH 100/0/0 to 0/100/0 to 0/99/1. The desired fractions were combined, evaporated under reduced pressure and coevaporated with toluene. The solids were dried under vacuum at 50 ° C to provide methyl 4- (3-amino-5-methoxyphenoxy) -2,2-dimethylbutanoate 2a (440 mg). Synthesis of intermediate 2b:
[0097] [0097] A mixture of 2-bromo-2- (4-chlorophenyl) -1- (6- (trifluoromethoxy) indolin-1-yl) ethanone 1c (1.57 g, 3.61 mmol), 4- (3 -amino- 5-methoxyphenoxy) -2,2-dimethylbutanoate methyl 2a (970 mg, 3.63 mmol) and diisopropylethylamine (961 µL, 5.58 mmol) in CH3CN (25 mL) was stirred at 55 ° C for 18 h. The mixture was allowed to reach room temperature and was poured into water (125 mL). The product was extracted (2x) with Et2O. The combined organic layers were dried over MgSO4, filtered and evaporated under reduced pressure. The residue was purified by flash chromatography on silica gel (40 g) using a gradient of heptane / EtOAc / EtOH 100/0/0 to 40/45/15. The desired fractions were combined and evaporated under reduced pressure and coevaporated with dioxane to provide 4- (3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl ) methyl) -5-methoxyphenoxy) -2-methylbutanoate 2b (2.24 g). Synthesis of Compound 2 and separation in Enantiomers 2A and 2B:
[0098] [0098] 1 M NaOH in water (9 mL, 9 mmol) was added to a stirring solution of 4- (3 - ((1- (4-chlorophenyl) -2-oxo-2- (6-
[0099] [0099] Compound 2 enantiomers (800 mg) were separated by preparative chiral SFC (Stationary phase: Chiralpak® Diacel AD 20 x 250 mm, mobile phase: CO2, EtOH + 0.4% iPrNH2). The product fractions were combined and evaporated under reduced pressure. The product eluted first was mixed with EtOAc (15 ml), water (5 ml) and 1 N HCl (1 ml). After stirring for 15 minutes, the layers were separated. The organic layer was dried over MgSO4, filtered, evaporated under reduced pressure and coevaporated with MeOH. The residue was triturated with water (4 ml) and MeOH (1.5 ml) while cooling in an ice bath. The solids were removed by filtration, washed (4x) with 4/1 H2O / MeOH and dried under vacuum at 45 ° C to provide Enantiomer 2A (292 mg). The second eluted product was mixed with EtOAc (15 ml), water (5 ml) and 1 N HCl (1 ml). After stirring for 30 minutes, the layers were separated. The organic layer was dried over MgSO4, filtered, evaporated under reduced pressure and coevaporated with MeOH. The residue was triturated with water (4 ml) and MeOH (1.5 ml) while cooling in an ice bath. The solids were removed by filtration, washed (4x) with 2/1 H2O / MeOH and dried under vacuum at 45 ° C to provide E nantiomer 2B (342 mg). Compound 2: 1
[00100] [00100] H NMR (400 MHz, DMSO-d6) į ppm 1.13 (d, J = 2.6 Hz, 6 H) 1.87 (t, J = 7.2 Hz, 2 H) 3.08 - 3.27 (m, 2 H) 3.61 (s, 3 H) 3.85 (t, J = 7.2 Hz, 2 H) 4.04 (td, J = 10.4, 7.2 Hz, 1 H) 4.52 (td, J = 10.2, 6.2 Hz, 1 H) 5.54 (d, J = 8.8 Hz, 1 H) 5.74 (t, J = 2 , 0 Hz, 1 H) 5.90 - 5.96 (m, 2 H) 6.43 (d, J = 8.6 Hz, 1 H) 7.01 (dd, J = 8.3, 1, 4 Hz, 1 H) 7.33 (d, J = 8.1 Hz, 1 H) 7.39 - 7.48 (m, 2 H) 7.50 - 7.60 (m, 2 H) 8, 03 (s, 1 H) 12.19 (ls, 1 H)
[00101] [00101] LC / MS (LC-A method): Rt 1.22 min, MH + 607 2A Enantiomer: 1
[00102] [00102] H NMR (400 MHz, DMSO-d6) į ppm 1.13 (d, J = 2.6 Hz, 6 H) 1.87 (t, J = 7.3 Hz, 2 H) 3.08 - 3.27 (m, 2 H) 3.61 (s, 3 H) 3.85 (t, J = 7.2 Hz, 2 H) 3.97 - 4.12 (m, 1 H) 4, 52 (td, J = 10.3, 6.5 Hz, 1 H) 5.54 (d, J = 8.6 Hz, 1 H) 5.74 (t, J = 2.1 Hz, 1 H) 5.90 - 5.96 (m, 2 H) 6.43 (d, J = 8.6 Hz, 1 H) 7.01 (dd, J = 8.1, 1.5 Hz, 1 H) 7 , 33 (d, J = 8.1 Hz, 1 H) 7.39 - 7.49 (m, 2 H) 7.55 (d, J = 8.6 Hz, 2 H) 8.03 (s, 1 H) 12.20 (ls, 1 H)
[00103] [00103] LC / MS (LC-A method): Rt 1.23 min, MH + 607
[00104] [00104] [Į] D20: + 49.6 ° (c 0.56, DMF)
[00105] [00105] Chiral SFC (SFC-B method): Rt 6.47 min, MH + 607 chiral purity 100%. 2B Enantiomer: 1
[00106] [00106] H NMR (400 MHz, DMSO-d6) į ppm 1.13 (d, J = 2.6 Hz, 6 H) 1.87 (t, J = 7.2 Hz, 2 H) 3.08 - 3.28 (m, 2 H) 3.61 (s, 3 H) 3.85 (t, J = 7.2 Hz, 2 H) 4.04 (td, J = 10.3, 7.3 Hz, 1 H) 4.52 (td, J = 10.3, 6.5 Hz, 1 H) 5.54 (d, J = 8.8 Hz, 1 H) 5.74 (t, J = 2 , 0 Hz, 1 H) 5.90 - 5.96 (m, 2 H) 6.43 (d, J = 8.6 Hz, 1 H) 7.01 (dd, J = 8.1, 1, 5 Hz, 1 H) 7.33 (d, J = 8.1 Hz, 1 H) 7.41 - 7.46 (m, 2 H) 7.55 (m, J = 8.6 Hz, 2 H ) 8.03 (s, 1 H) 12.20 (ls, 1 H)
[00107] [00107] LC / MS (LC-A method): Rt 1.23 min, MH + 607
[00108] [00108] [Į] D20: -49.2 ° (c 0.445, DMF)
[00109] [00109] Chiral SFC (SFC-B method): Rt 7.18 min, MH + 607 chiral purity 98.8%. Example 3: 4- (3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) -2 acid synthesis , 2-dimethylbutanoic (Compound 3) Synthesis of intermediate 3a:
[00110] [00110] 5-Oxaspospiro [2.4] heptan-4-one (930 mg, 8.29 mmol) was mixed with a solution of 33% HBr in AcOH (8 mL). The reaction mixture was stirred at room temperature for 1.5 h and poured into ice water (50 ml). After stirring for 10 min, the precipitate was filtered off, washed (5x) with water and dried under vacuum at 45 ° C to provide 1- (2-bromoethyl) cyclopropane-1-carboxylic acid 3a (753 mg). Synthesis of intermediate 3b:
[00111] [00111] A solution of 1- (2-bromoethyl) cyclopropane-1-carboxylic acid 3a (540 mg, 2.8 mmol) in MeOH (11 mL) was stirred under N2 atm while cooling in an ice bath. Thionyl chloride (304 µL, 4.2 mmol) was added dropwise. The reaction mixture was stirred at room temperature for 40 h. The solvents were evaporated under reduced pressure and coevaporated with CH3CN to provide methyl 1- (2-bromoethyl) cyclopropane-1-carboxylate 3b (380 mg). Synthesis of intermediate 3c:
[00112] [00112] To a stirred solution of methyl 1- (2-bromoethyl) cyclopropane-1-carboxylate 3b (380 mg, 1.84 mmol) in DMF (10 mL) were added 3-amino-5-methoxyphenol [CAS 162155 -27-3] (250 mg, 1.80 mmol) and Cs2CO3 (1.17 g, 3.59 mmol). The reaction was stirred at 60 ° C for 18 h, and allowed to reach room temperature. The mixture was poured into H2O (60 ml). The product was extracted (2x) with Et2O. The combined organic layers were washed with brine, dried over MgSO4, filtered, and the solvent was evaporated under reduced pressure and coevaporated with toluene. The residue was purified by flash chromatography on silica gel (12 g) using a gradient from heptane / CH2Cl2 / MeOH 100/0/0 to 0/100/0 to 0/99/1. The product fractions were combined, evaporated under reduced pressure and coevaporated with CH3CN to provide methyl 1- (2- (3-amino-5-methoxyphenoxy) ethyl) cyclopropane-1-carboxylate 3c (220 mg). Synthesis of intermediate 3d:
[00113] [00113] A mixture of 2-bromo-2- (4-chlorophenyl) -1- (6- (trifluoromethoxy) indolin-1-yl) ethanone 1c (320 mg, 0.736 mmol), 1- (2- (3- methyl amino-5-methoxyphenoxy) ethyl) cyclopropane-1-carboxylate 3c (220 mg, 0.829 mmol) and diisopropylethylamine (254 µL, 1.47 mmol) in 2-butanol (7.5 mL) was stirred at 55 ° C for 16 h. The mixture was allowed to reach room temperature and was poured into water (25 ml). The product was extracted (2x) with Et2O. The combined organic layers were washed with brine, dried over MgSO4, filtered and evaporated under reduced pressure. The residue was purified by flash chromatography on silica gel (12 g) using a gradient of heptane / EtOAc / EtOH 100/0/0 to 60/30/10. The product fractions were combined and evaporated under reduced pressure and coevaporated with dioxane to provide 1- (2- (3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1 -yl) ethyl) amino) -5-methoxyphenoxy) ethyl) cyclopropane-methyl 1-carboxylate 3d (456 mg). Synthesis of Compound 3:
[00114] [00114] 1 M NaOH in water (1.84 mL, 1.84 mmol) was added to a stirring solution of 1- (2- (3 - ((1- (4-chlorophenyl) -2-oxo- 2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) ethyl) cyclopropane-1-methyl carboxylate 3d (0.456 mg, 0.737 mmol) in dioxane (3 ml). The reaction mixture was stirred at room temperature under N2 atm for 42 h. Water (15 ml) and 1 N HCl (2 ml) were added. After stirring for 10 min, the precipitate was filtered off, washed (3 x) with water and dried under vacuum at 45 ° C. The residue was purified by preparative HPLC (Stationary phase: RP XBridge® Prep C18 OBD - 10 µm, 30 x 150 mm, mobile phase: 0.25% NH4HCO3 solution in water, CH3CN). The product fractions were combined and the organic solvents were evaporated. The remaining aqueous solution was extracted (2x) with Et2O. The combined organic layers were washed with brine, dried over MgSO4, filtered, evaporated under reduced pressure and coevaporated with MeOH. The resulting foam was stirred in 3/1 H2O / MeOH (4 mL), filtered off, washed (3 x) with 3/1 H2O / MeOH and dried under vacuum at 45 ° C to provide 1- (2- (3 - (((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) ethyl) cyclopropane-1-carboxylic (Compound 3, 255 mg) as a racemic mixture. Compound 3: 1
[00115] [00115] H NMR (400 MHz, DMSO-d6) į ppm 0.73 - 0.84 (m, 2 H) 1.00 - 1.11 (m, 2 H) 1.81 - 1.91 (m , 2 H) 3.07 - 3.26 (m, 2 H) 3.62 (s, 3 H) 3.97 (t, J = 7.3 Hz, 2 H) 4.05 (td, J = 10.3, 7.3 Hz, 1 H) 4.52 (td, J = 10.2, 6.6 Hz, 1 H) 5.55 (d, J = 8.6 Hz, 1 H) 5, 77 (t, J = 2.1 Hz, 1 H) 5.90 - 5.98 (m, 2 H) 6.42 (d, J = 8.6 Hz, 1 H) 7.00 (dd, J = 8.1, 1.5 Hz, 1 H) 7.33 (d, J = 8.1 Hz, 1 H) 7.39 - 7.49 (m, 2 H) 7.55 (d, J = 8.4 Hz, 2 H) 8.03 (s, 1 H) 12.19 (ls, 1 H)
[00116] [00116] LC / MS (LC-B method): Rt 2.19 min, MH + 605 Example 4: 4- (3 - ((1- (4-chlorophenyl) -2-oxo-2- (6 - (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) -3-methylbutanoic (Compound 4) and separation in Stereoisomers 4A, 4B, 4C and 4D Synthesis of intermediate 4a:
[00117] [00117] To a stirred solution of ethyl 4-bromo-3-methylbutanoate [CAS 56703-10-7] (1.0 g, 4.78 mmol) in DMF (15 mL) were added 3-amino-5- methoxyphenol [CAS 162155-27-3] (666 mg, 4.78 mmol) and Cs2CO3 (3.12 g, 9.57 mmol). The reaction was stirred at 70 ° C for 16 h, and allowed to reach room temperature. The mixture was poured into H2O (75 ml). The product was extracted (2x) with Et2O. The combined organic layers were washed with brine, dried over MgSO4, filtered, and the solvent was evaporated under reduced pressure. The residue was purified by flash chromatography on silica gel (25 g) using a 100/0 to 50/50 heptane / EtOAc gradient. The product fractions were combined, evaporated under reduced pressure and coevaporated with CH3CN, yielding ethyl 4- (3-amino-5-methoxyphenoxy) -3-methylbutanoate 4a (430 mg). Synthesis of intermediate 4b:
[00118] [00118] A mixture of 2-bromo-2- (4-chlorophenyl) -1- (6- (trifluoromethoxy) indolin-1-yl) ethanone 1c (430 mg, 1.15 mmol), 4- (3- amino Ethyl-5-methoxyphenoxy) -3-methylbutanoate 4a (430 mg, 1.61 mmol) and diisopropylethylamine (396 µL, 2.30 mmol) in CH3CN (15 mL) was stirred at 60 ° C for 18 h under N 2 atmosphere. The mixture was allowed to reach room temperature, and was poured into water (75 mL). The product was extracted (2x) with Et2O. The combined organic layers were washed with brine, dried over MgSO4, filtered and evaporated under reduced pressure. The residue was purified by flash chromatography on silica gel (12 g) using a gradient of heptane / EtOAc / EtOH 100/0/0 to 40/45/15. The product fractions were combined and evaporated under reduced pressure and coevaporated with EtOH to provide 4- (3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) -3-methylbutanoate 4b (714 mg). Synthesis of Compound 4 and separation in Stereoisomers 4A, 4B, 4C and 4D:
[00119] [00119] 1 M NaOH in water (2.9 mL, 2.9 mmol) was added to a stirring solution of 4- (3 - ((1- (4-chlorophenyl) -2-oxo-2- ( 6-
[00120] [00120] The 4 stereoisomers of Compound 4 (274 mg) were separated by preparative chiral SFC (Stationary phase: Chiralpak® Diacel AD 20 x 250 mm, mobile phase: CO2, iPrOH + 0.4% iPrNH2). The product fractions were combined and evaporated under reduced pressure. The stereoisomers in the product fractions of the second and third eluted peaks were not completely separated and required additional separation through preparative chiral SFC (Stationary phase: Chiralpak® Diacel AD 20 x 250 mm, mobile phase: CO2, EtOH + iPrNH2 at 0, 4%). The product fractions were combined and evaporated under reduced pressure. The 4 stereoisomers were solidified by lyophilization from a solvent mixture of CH3CN and water to provide Stereoisomers 4A (72 mg), 4B (35 mg), 4C (35 mg) and 4D (67 mg). Compound 4: 1
[00121] [00121] H NMR (400 MHz, DMSO-d6) į ppm 0.96 (d, J = 6.6 Hz, 3 H)
[00122] [00122] LC / MS (LC-B method): Rt 2.07 min, MH + 593 Stereoisomer 4A: 1
[00123] [00123] H NMR (400 MHz, DMSO-d6) į ppm 0.96 (d, J = 6.8 Hz, 3 H) 2.05 - 2.13 (m, 1 H) 2.22 (dq, J = 13.1, 6.6 Hz, 1 H) 2.33 - 2.40 (m, 1 H) 3.06 - 3.21 (m, 2 H) 3.62 (s, 3 H) 3 , 65 - 3.76 (m, 2 H) 4.05 (td, J = 10.4, 7.2 Hz, 1 H) 4.52 (td, J = 10.1, 6.4 Hz, 1 H) 5.56 (d, J = 8.8 Hz, 1 H) 5.76 (t, J = 2.0 Hz, 1 H) 5.93 (t, J = 1.9 Hz, 1 H) 5.97 (t, J = 1.7 Hz, 1 H) 6.43 (d, J = 8.6 Hz, 1 H) 7.00 (dd, J = 8.1, 1.5 Hz, 1 H) 7.33 (d, J = 8.1 Hz, 1 H) 7.43 (m, J = 8.6 Hz, 2 H) 7.55 (m, J = 8.6 Hz, 2 H) 8.03 (ls, 1 H) 11.51 (ls, 1 H)
[00124] [00124] LC / MS (LC-B method): Rt 2.04 min, MH + 593
[00125] [00125] [Į] D20: -59.6 ° (c 0.245, DMF)
[00126] [00126] Chiral SFC (SFC-C method): Rt 5.84 min, MH + 593 100% chiral purity. Stereoisomer 4B: 1
[00127] [00127] H NMR (400 MHz, DMSO-d6) į ppm 0.96 (d, J = 6.6 Hz, 3 H) 2.07 - 2.14 (m, 1 H) 2.22 (dq, J = 13.2, 6.5 Hz, 1 H) 2.38 (dd, J = 15.2, 5.5 Hz, 1 H) 3.02 - 3.23 (m, 2 H) 3.62 (s, 3 H) 3.65 - 3.76 (m, 2 H) 4.04 (td, J = 10.3, 7.3 Hz, 1 H) 4.52 (td, J = 10.5 , 6.2 Hz, 1 H) 5.56 (d, J = 8.8 Hz, 1 H) 5.76 (t, J = 2.0 Hz, 1 H) 5.93 (t, J = 1 , 8 Hz, 1 H) 5.97 (t, J = 1.7 Hz, 1 H) 6.43 (d, J = 8.6 Hz, 1 H) 7.00 (dd, J = 8.0 , 1.4 Hz, 1 H) 7.33 (d, J = 8.1 Hz, 1 H) 7.41 - 7.46 (m, 2 H) 7.55 (m, J = 8.4 Hz , 2 H) 8.03 (ls, 1 H) 12.00 (ls, 1 H)
[00128] [00128] LC / MS (LC-B method): Rt 2.04 min, MH + 593
[00129] [00129] [Į] D20: -47.5 ° (c 0.255, DMF)
[00130] [00130] Chiral SFC (SFC-C method): Rt 6.34 min, MH + 593 chiral purity 98.0%. Stereoisomer 4C: 1
[00131] [00131] H NMR (400 MHz, DMSO-d6) į ppm 0.96 (d, J = 6.6 Hz, 3 H) 2.06 - 2.14 (m, 1 H) 2.22 (dq, J = 13.3, 6.4 Hz, 1 H) 2.37 (dd, J = 15.3, 5.6 Hz, 1 H) 3.08 - 3.22 (m, 2 H) 3.62 (s, 3 H) 3.66 - 3.74 (m, 2 H) 4.04 (td, J = 10.5, 7.0 Hz, 1 H) 4.52 (td, J = 10.2 , 6.6 Hz, 1 H) 5.56 (d, J = 8.8 Hz, 1 H) 5.76 (t, J = 2.1 Hz, 1 H) 5.93 (t, J = 1 , 7 Hz, 1 H) 5.97 (t, J = 1.8 Hz, 1 H) 6.44 (d, J = 8.8 Hz, 1 H) 7.01 (dd, J = 8.1 , 1.5 Hz, 1 H) 7.33 (d, J = 8.1 Hz, 1 H) 7.44 (m, J = 8.6 Hz, 2 H) 7.55 (m, J = 8 , 6 Hz, 2 H) 8.03 (ls, 1 H) 10.85 - 12.62 (m, 1 H)
[00132] [00132] LC / MS (LC-B method): Rt 2.04 min, MH + 593
[00133] [00133] [Į] D20: + 47.7 ° (c 0.26, DMF)
[00134] [00134] Chiral SFC (SFC-C method): Rt 6.31 min, MH + 593 chiral purity 100%. 4D Stereoisomer: 1
[00135] [00135] H NMR (360 MHz, DMSO-d6) į ppm 0.96 (d, J = 7.0 Hz, 3 H) 2.06 - 2.15 (m, 1 H) 2.17 - 2, 28 (m, 1 H) 2.38 (dd, J = 15.4, 5.5 Hz, 1 H) 3.07 - 3.26 (m, 2 H) 3.62 (s, 3 H) 3 , 65 - 3.76 (m, 2 H) 4.05 (td, J = 10.2, 7.3 Hz, 1 H) 4.52 (td, J = 10.3, 6.4 Hz, 1 H) 5.57 (d, J = 8.8 Hz, 1 H) 5.76 (t, J = 2.0 Hz, 1 H) 5.93 (t, J = 1.5 Hz, 1 H) 5.97 (t, J = 1.5 Hz, 1 H) 6.46 (d, J = 8.4 Hz, 1 H) 7.01 (dd, J = 8.2, 1.6 Hz, 1 H) 7.33 (d, J = 8.4 Hz, 1 H) 7.44 (m, J = 8.4 Hz, 2 H) 7.55 (m, J = 8.4 Hz, 2 H) 8.03 (ls, 1 H) 11.95 (ls, 1 H)
[00136] [00136] LC / MS (LC-B method): Rt 2.04 min, MH + 593
[00137] [00137] [Į] D20: + 60.7 ° (c 0.285, DMF)
[00138] [00138] Chiral SFC (SFC-C method): Rt 7.58 min, MH + 593 100% chiral purity. Example 5: synthesis of 2- (1 - ((3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoro-methoxy) indolin-1-yl) ethyl) amino) - 5- methoxyphenoxy) methyl) cyclopropyl) acetic (Compound 5) and chiral separation in Enantiomers 5A and 5B Synthesis of intermediate 5a:
[00139] [00139] Methyl 2- (1- (Bromomethyl) cyclopropyl) acetate [855473-50-6] (306 mg, 1.478 mmol) was added dropwise to a solution of 3-methoxy-5-nitrophenol [7145-49 -5] (250 mg, 1.478 mmol) and K2CO3 (306 mg, 2.217 mmol) in DMF (2.5 mL). The mixture was stirred at 60 ° C for 16 h. The reaction was cooled to 0 ° C and diluted with water and ice. The mixture was extracted with EtOAc. The organic layer was washed with water, dried over MgSO4, filtered and the solvent was concentrated under reduced pressure to give methyl 2- (1 - ((3-methoxy-5-nitrophenoxy) methyl) cyclopropyl) acetate 5a. The yield was considered to be quantitative. The compound was used as such in the next step. Synthesis of intermediate 5b:
[00140] [00140] A solution of 2- (1 - ((3-methoxy-5-nitrophenoxy) methyl) cyclopropyl) methyl acetate 5a (520 mg, 1,761 mmol) in MeOH (8 mL), containing a catalytic amount of Pd / 10% C (300 mg, 0.282 mmol), was hydrogenated under atmospheric pressure of H2 at room temperature for 18 h. The catalyst was removed by filtration over a short pad of Celite® and the filter cake was rinsed several times with EtOAc. The combined filtrates were evaporated to give 2- (1 - ((3-amino-5-
[00141] [00141] A mixture of 2-bromo-2- (4-chlorophenyl) -1- (6- (trifluoromethoxy) indolin-1-yl) ethanone 1c (250 mg, 0.575 mmol), 2- (1 - ((3 - amino-5-methoxy-phenoxy) methyl) cyclopropyl) methyl acetate 5b (184 mg, 0.575 mmol) and diisopropylethylamine (200 µL, 1.15 mmol) in CH3CN (7.5 mL) was stirred at 60 ° C for 18 h. The solvent was concentrated under reduced pressure. Water / ice was added and the mixture was extracted with EtOAc. The organic layer was washed with water, dried over MgSO4, filtered and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (30 µm, 24 g, heptane / EtOAc 75/25). The product fractions were combined and evaporated under reduced pressure. The residue was crystallized from CH3CN / diisopropyl ether and dried to give 2- (1 - ((3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoro-methoxy ) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) methyl) cyclopropyl) methyl acetate 5c (206 mg). Synthesis of Compound 5 and separation in Enantiomers 5A and 5B:
[00142] [00142] LiOH monohydrate (63 mg, 1.502 mmol) in water (1.63 mL) was added dropwise to a solution of 2- (1 - ((3 - ((1- (4-chlorophenyl) - 2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) methyl) cyclopropyl) methyl acetate 5c (186 mg, 0.3 mmol) in THF (3.7 mL). The reaction mixture was stirred at room temperature for 24 h. 3 N HCl was added to acidify the reaction mixture, and the aqueous solution was extracted with EtOAc. The combined organic layers were dried over MgSO4, filtered and concentrated under reduced pressure. The residue was crystallized from CH3CN / diisopropyl ether and dried to give 2- (1 - ((3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) acid indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) methyl) cyclopropyl) acetic (Compound 5, 115 mg).
[00143] [00143] The separation of the Compound 5 enantiomers (71 mg) was performed by preparative chiral SFC (Stationary phase: Chiralpak® Diacel AD 20 x 250 mm, mobile phase: CO2, EtOH + 0.4% iPrNH2). For both enantiomers, the product fractions were combined and evaporated under reduced pressure. The residues were divided between water and Et2O. The mixtures were acidified to pH 1-2 by the addition of 1 N HCl and the layers were separated. The aqueous layer was extracted again with Et2O. The combined organic layers were dried over MgSO4, filtered and evaporated under reduced pressure. The residues were dried in vacuo at 50 ° C to provide E nantiomer 5A (22 mg) and Enantiomer 5B (23 mg) as off-white powders. Compound 5: 1
[00144] [00144] H NMR (500 MHz, DMSO-d6) į ppm 0.49 - 0.55 (m, 4 H) 2.33 (s, 2 H) 2.99 - 3.25 (m, 2 H) 3.62 (s, 3 H) 3.73 (s, 2 H) 4.05 (td, J = 10.40, 7.25 Hz, 1 H) 4.52 (td, J = 10.25, 6.31 Hz, 1 H) 5.57 (d, J = 9.14 Hz, 1 H) 5.74 (s, 1 H) 5.92 (s, 1 H) 5.95 (s, 1 H ) 6.45 (d, J = 9.14 Hz, 1 H) 7.02 (dl, J = 9.14 Hz, 1 H) 7.34 (d, J = 8.20 Hz, 1 H) 7 , 44 (d, J = 8.20 Hz, 2 H) 7.55 (d, J = 8.51 Hz, 3 H) 8.03 (s, 1 H) 12.01 (sl, 1 H)
[00145] [00145] LC / MS (LC-C method): Rt 3.18 min, MH + 605
[00146] [00146] PF = 111 ° C 5A enantiomer: 1
[00147] [00147] H NMR (360 MHz, DMSO-d6) į ppm 0.46 - 0.58 (m, 4 H) 2.33 (s, 2 H) 3.04 - 3.27 (m, 2 H) 3.61 (s, 3 H) 3.72 (s, 2 H) 4.05 (td, J = 10.2, 7.3 Hz, 1 H) 4.52 (td, J = 10.2, 6.6 Hz, 1 H) 5.57 (d, J = 9.1 Hz, 1 H) 5.74 (t, J = 2.0 Hz, 1 H) 5.92 (t, J = 1, 6 Hz, 1 H) 5.94 - 5.97 (m, 1 H) 6.45 (d, J = 8.8 Hz, 1 H) 7.01 (dd, J = 8.1, 1.5 Hz, 1 H) 7.33 (d, J = 8.4 Hz, 1 H) 7.44 (d, J = 8.4 Hz, 2 H) 7.51 - 7.59 (m, 2 H) 8.03 (s, 1 H) 12.12 (ls, 1 H)
[00148] [00148] LC / MS (LC-A method): Rt 1.15 min, MH + 605
[00149] [00149] [Į] D20: + 37.0 ° (c 0.135, DMF)
[00150] [00150] Chiral SFC (SFC-E method): Rt 5.84 min, MH + 605 chiral purity 100%. 5B Enantiomer: 1
[00151] [00151] H NMR (360 MHz, DMSO-d6) į ppm 0.46 - 0.57 (m, 4 H) 2.33 (s, 2 H) 3.05 - 3.26 (m, 2 H) 3.61 (s, 3 H) 3.72 (s, 2 H) 4.05 (td, J = 10.4, 7.3 Hz, 1 H) 4.52 (td, J = 10.3, 6.4 Hz, 1 H) 5.57 (d, J = 8.8 Hz, 1 H) 5.74 (t, J = 2.0 Hz, 1 H) 5.92 (t, J = 1, 8 Hz, 1 H) 5.94 - 5.97 (m, 1 H) 6.45 (d, J = 8.8 Hz, 1 H) 7.01 (dd, J = 8.2, 1.6 Hz, 1 H) 7.33 (d, J = 8.1 Hz, 1 H) 7.44 (d, J = 8.4 Hz, 2 H) 7.50 - 7.61 (m, 2 H) 8.03 (s, 1 H) 12.11 (ls, 1 H)
[00152] [00152] LC / MS (LC-A method): Rt 1.15 min, MH + 605
[00153] [00153] [Į] D20: -48.8 ° (c 0.16, DMF)
[00154] [00154] Chiral SFC (SFC-E method): Rt 6.53 min, MH + 605 chiral purity 100%. Example 6A: synthesis of (1R *, 2R *) - 2 - ((3 - ((1- (4-chlorophenyl) -2-oxo- 2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) methyl) - cyclopropanecarboxylic (Compound 6A) and separation in Stereoisomers 6AA and 6AB Synthesis of intermediate 6a:
[00155] [00155] To a solution of ethyl 2-formylcyclopropanecarboxylate [20417-61-2] (9 mL, 67.996 mmol) in MeOH (200 mL) was added portion by portion NaBH4 (5.15 g, 133.993 mmol) at 0 ° C and the mixture was stirred at 0 ° C for 2 h. CH 2Cl2 and water were added. The layers were separated; the aqueous layer was extracted with CH2Cl2 and the combined organic layers were dried over MgSO4, filtered and concentrated under reduced pressure to give ethyl 2- (hydroxymethyl) cyclopropanecarboxylate 6a (9.15 g). The compound was used as such in the next step. Synthesis of intermediate 6b:
[00156] [00156] Di-tert-butyl azodicarboxylate (4.8 g, 20.809 mmol) was added portion by portion to a solution of 3-methoxy-5-nitrophenol [7145-49-5] (3.2 g, 18.917 mmol ), Ethyl 2- (hydroxymethyl) cyclopropanecarboxylate 6a (3 g, 20.809 mmol) and PPh3 (5.46 g, 20.809 mmol) in THF (150 mL). The reaction was stirred at room temperature under N2 for 18 h. The solution was concentrated under reduced pressure. The crude residue was purified by preparative LC (20-45 µm irregular SiOH, 220 g, 85/15 to 75/25 heptane / EtOAc). The pure fractions were combined and concentrated under reduced pressure to give methyl 2 - ((3-methoxy-5-nitrophenoxy) methyl) cyclopropanecarboxylate 6b (1.4 g). Synthesis of intermediate 6c and chiral separation in enantiomers 6d and 6e:
[00157] [00157] A solution of methyl 2 - ((3-methoxy-5-nitrophenoxy)) cyclopropanecarboxylate 6b (1.3 g, 4.402 mmol) in EtOH (65 mL) containing a catalytic amount of 10% Pd / C (750 mg, 0.704 mmol) was hydrogenated under atmospheric pressure of H2 at room temperature for 4 h. The catalyst was removed by filtration over a short pad of Celite® and the filter cake was rinsed several times with EtOAc. The combined filtrates were evaporated. The crude residue was purified by preparative LC (Irregular SiOH 20-45 µm, 40 g, heptane / EtOAc 80/20). The pure fractions were combined and the solvent was evaporated to dryness to give methyl 2 - ((3-amino-5-methoxyphenoxy) methyl) cyclopropanecarboxylate 6c (780 mg). The enantiomers were separated by chiral SFC (Stationary phase: Chiralpak® AD-H 5 µm 250 x 30 mm, mobile phase: 80% CO2, 20% EtOH) to give the first eluted enantiomer 6d (trans R * , R *, 344 mg, [Į] D20: -78.6 ° (c 0.257, DMF)) and the second eluted enantiomer 6e (trans S *, S *, 371 mg, [Į] D20: +74 , 5 ° (c 0.251, DMF)). Synthesis of intermediate 6f:
[00158] [00158] A mixture of 2-bromo-2- (4-chlorophenyl) -1- (6- (trifluoromethoxy) indolin-1-yl) ethanone 1c (376 mg, 0.864 mmol), 2 - ((3- amino- 5- methoxyphenoxy) methyl) cyclopropanecarboxylatomethoxyphenoxy) methyl) cyclopropyl l) acetate (1R *, 2R *) - methyl 6d (344 mg, 1.297 mmol) and diisopropylethylamine (298 µL, 1.729 mmol) in CH3CN (12 mL) was stirred at 80 ° C for 5 days. The solvent was concentrated under reduced pressure. Water / ice was added and the mixture was extracted with EtOAc. The organic layer was washed with water, dried over MgSO4, filtered and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (25-30 µm, 40 g, heptane / EtOAc 80/20). The product fractions were combined and evaporated under reduced pressure to give 2 - ((3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethyl) indolin-1-yl) ethyl) amino) ) (5R-methoxyphenoxy) methyl) -cyclopropanecarboxylate of (1R *, 2R *) - methyl 6f (500 mg). Synthesis of Compound 6A and chiral separation in Stereoisomers 6AA and 6AB:
[00159] [00159] LiOH monohydrate (169 mg, 4.039 mmol) in water (10 mL) was added dropwise to a solution of 2 - ((3 - ((1- (4-chlorophenyl) -2-oxo-2 - (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) methyl) (1R *, 2R *) - cyclopropanecarboxylate - methyl 6f (500 mg, 0.808 mmol) in THF (10 mL ). The reaction mixture was stirred at room temperature for 18 h and at 45 ° C for 5 h. 3 N HCl was added to acidify the solution, and the mixture was extracted with EtOAc. The combined organic layers were dried over MgSO4, filtered and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (20-45 µm, 40 g, heptane / EtOAc 80/20). The product fractions were combined and evaporated under reduced pressure to give acid (1R *, 2R *) - 2 - ((3 - ((1- (4-chlorophenyl) -2-oxo- 2- (6- (trifluoromethyl) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) methyl) cyclopropanecarboxylic (Compound 6A, 780 mg). The two stereoisomers were separated by chiral SFC (Stationary phase: Chiralcel® OD-H 5 µm 250 x 30 mm, mobile phase: 55% CO2, 45% iPrOH) to give, after solidification from heptane / ether of diisopropyl, the Stereoisomer eluted first 6AA (123 mg) and the Stereoisomer eluted second 6A (125 mg). Stereoisomer 6AA: 1
[00160] [00160] H NMR (500 MHz, DMSO-d6) į ppm 0.75 - 0.96 (m, 1 H) 1.00 - 1.07 (m, 1 H) 1.44 - 1.57 (m , 1 H) 1.57 - 1.70 (m, 1 H) 3.09 - 3.27 (m, 2 H) 3.62 (s, 3 H) 3.69 (d ld, J = 10, 25, 7.72 Hz, 1 H) 3.80 - 3.95 (m, 1 H) 4.00 - 4.09 (m, 1 H) 4.39 - 4.65 (m, 1 H) 5 , 57 (dl, J = 8.51 Hz, 1 H) 5.76 (s, 1 H) 5.95 (s, 1 H) 5.97 (s, 1 H) 6.46 (dl, J = 8.83 Hz, 1 H) 7.02 (dl, J = 8.20 Hz, 1 H) 7.34 (d, J = 8.20 Hz, 1 H) 7.44 (d, J = 8, 51 Hz, 2 H) 7.55 (dl, J = 8.20 Hz, 2 H) 8.04 (ls, 1 H) 12.02 (ls, 1 H)
[00161] [00161] LC / MS (LC-C method): Rt 2.95 min, MH + 591
[00162] [00162] [Į] D20: -78.0 ° (c 0.282, DMF)
[00163] [00163] Chiral SFC (SFC-G method): Rt 1.08 min, MH + 591 chiral purity 99.82%. Stereoisomer 6AB: 1
[00164] [00164] H NMR (500 MHz, DMSO-d6) į ppm 0.79 - 0.90 (m, 1 H) 1.00 - 1.08 (m, 1 H) 1.43 - 1.57 (m , 1 H) 1.57 - 1.72 (m, 1 H) 2.95 - 3.27
[00165] [00165] LC / MS (LC-C method): Rt 2.95 min, MH + 591
[00166] [00166] [Į] D20: + 12.9 ° (c 0.272, DMF)
[00167] [00167] Chiral SFC (SFC-G method): Rt 1.87 min, MH + 591 chiral purity 99.55%. Example 6B: Synthesis of ((1S *, 2S *) - 2 - ((3 - ((1- (4-chlorophenyl) -2-oxo- 2- (6- (trifluoromethoxy) indolin-1-yl) ethyl ) amino) -5-methoxyphenoxy) methyl) - cyclopropanecarboxylic (Compound 6B) and separation in Stereoisomers 6BA and 6BB Synthesis of intermediate 6g:
[00168] [00168] A mixture of 2-bromo-2- (4-chlorophenyl) -1- (6- (trifluoromethoxy) indolin-1-yl) ethanone 1c (405 mg, 0.932 mmol), 2 - ((3- amino- 5- methoxyphenoxy) methyl) cyclopropanecarboxylatomethoxyphenoxy) methyl) cyclopropyl l) (1S *, 2S *) - methyl 6e acetate (371 mg, 1.398 mmol) and diisopropylethylamine (321 µL, 1.864 mmol) in CH3CN (12 mL) was stirred at 80 ° C for 48 h. The solvent was concentrated under reduced pressure. Water / ice was added and the mixture was extracted with EtOAc. The organic layer was washed with water, dried over MgSO4, filtered and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (25-30 µm, 40 g, heptane / EtOAc 80/20). The product fractions were combined and evaporated under reduced pressure to give 2 - ((3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethyl) indolin-1-yl) ethyl) amino) ) -5-methoxyphenoxy) methyl) -cyclopropanecarboxylate (1S *, 2S *) - methyl 6g (580 mg). Synthesis of Compound 6B and chiral separation in Stereoisomers 6BA and 6BB:
[00169] [00169] LiOH monohydrate (203 mg, 4.846 mmol) in water (10 mL) was added dropwise to a solution of 2 - ((3 - ((1- (4-chlorophenyl) -2-oxo-2 - (6 (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) methyl) cyclopropanecarboxylate (1S *, 2S *) - methyl 6g (600 mg, 0.969 mmol) in THF (10 mL). The reaction mixture was stirred at room temperature for 7 h. 3 N HCl was added to acidify the solution, and the mixture was extracted with EtOAc. The combined organic layers were dried over MgSO4, filtered and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (20-45 µm, 40 g, heptane / EtOAc 80/20). The product fractions were combined and evaporated under reduced pressure to give acid (1S *, 2S *) - 2 - ((3 - ((1- (4-chlorophenyl) -2-oxo- 2- (6- (trifluoromethyl) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) methyl) cyclopropanecarboxylic (Compound 6B, 348 mg). The two stereoisomers were separated by chiral SFC (Stationary phase: Chiralcel® OD-H 5 µm 250 x 30 mm, mobile phase: 55% CO2, 45% iPrOH) to give, after solidification from heptane / ether of diisopropyl, the Stereoisomer eluted first 6BA (109 mg) and the Stereoisomer eluted second 6BB (102 mg). Stereoisomer 6BA: 1
[00170] [00170] H NMR (500 MHz, DMSO-d6) į ppm 0.73 - 0.98 (m, 1 H) 0.98 - 1.08 (m, 1 H) 1.49 - 1.58 (m , 1 H) 1.58 - 1.71 (m, 1 H) 3.00 - 3.27 (m, 2 H) 3.61 (s, 3 H) 3.68 (dd, J = 10.40 , 7.57 Hz, 1 H) 3.84 (dd, J = 10.56, 6.15 Hz, 1 H) 4.00 - 4.08 (m, 1 H) 4.50 - 4.57 ( m, 1 H) 5.57 (d, J = 8.83
[00171] [00171] LC / MS (LC-D method): Rt 2.82 min, MH + 591
[00172] [00172] [Į] D20: -12.5 ° (c 0.28, DMF)
[00173] [00173] Chiral SFC (SFC-G method): Rt 1.10 min, without MH +, 100% chiral purity. Stereoisomer 6BB: 1
[00174] [00174] H NMR (500 MHz, DMSO-d6) į ppm 0.78 - 0.95 (m, 1 H) 1.00 -1.08 (m, 1H) 1.48 - 1.59 (m, 1 H) 1.59 - 1.68 (m, 1 H) 2.91 - 3.25 (m, 2 H) 3.61 (s, 3 H) 3.68 (d ld, J = 10.40 , 7.57 Hz, 1 H) 3.84 (dd, J = 10.56, 6.15 Hz, 1 H) 4.00 - 4.09 (m, 1 H) 4.30 - 4.58 ( m, 1 H) 5.57 (d, J = 8.83 Hz, 1 H) 5.75 (s, 1 H) 5.95 (s, 1 H), 5.96 (s, 1H) 6, 45 (dl, J = 8.83 Hz, 1 H) 7.01 (dl, J = 7.88 Hz, 1 H) 7.33 (d, J = 8.20 Hz, 1 H) 7.44 ( d, J = 8.20 Hz, 2 H) 7.55 (d, J = 8.20 Hz, 2 H) 8.03 (s, 1 H) 12.11 (ls, 1 H)
[00175] [00175] LC / MS (LC-D method): Rt 2.81 min, MH + 591
[00176] [00176] [Į] D20: + 81.4 ° (c 0.28, DMF)
[00177] [00177] Chiral SFC (SFC-G method): Rt 1.87 min, without MH +, chiral purity 99.02%. Example 7: synthesis of (1r, 3r) -3- (3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) - 5-methoxyphenoxy) cyclobutene carboxylic (Compound 7) and separation into Enantiomers 7A and 7B
[00178] [00178] Under a flow of N2, to a solution of ethyl 3-hydroxycyclobutanecarboxylate [17205-02-6] (1 g, 6.936 mmol) in CH2Cl2 (30 mL) were added pyridine (0.838 mL) and tosyl anhydride ( 2.49 g, 7.63 mmol). The mixture was stirred overnight at room temperature. The mixture was concentrated in vacuo, suspended in diethyl ether (200 ml) and washed with 0.5 M hydrochloric acid (2 × 60 ml), a saturated solution of sodium hydrogen carbonate (2 × 60 ml), water (60 ml) and brine (50 ml). The solution was dried over sodium sulfate, filtered and concentrated under reduced pressure to give ethyl 3- (tosyloxy) cyclobutanecarboxylate 7a (2.0 g). Synthesis of intermediate 7b:
[00179] [00179] Ethyl 3- (Tosyloxy) cyclobutanecarboxylate 7a (1.94 g, 6.504 mmol) was added dropwise to a mixture of 3-methoxy-5-nitrophenol [7145-49-5] (1.0 g, 5.912 mmol) and K2CO3 (981 mg, 7.095 mmol) in DMF (10 mL). The mixture was stirred at 80 ° C for 16 h. The mixture was diluted with water and extracted with EtOAc. The organic layer was washed with water, dried over MgSO4, filtered and the solvent was evaporated under reduced pressure. Purification was performed by flash chromatography on silica gel (15-40 µm, 80 g, heptane / EtOAc 95/5 to 85/15). The pure fractions were combined and evaporated to dryness to give 3- (3-methoxy-5-nitrophenoxy) cyclobutanecarboxylate of (1r, 3r) -ethyl 7b (1.1 g). Synthesis of intermediate 7c:
[00180] [00180] A solution of 3- (3-methoxy-5-nitrophenoxy) (1r, 3r) -ethyl 7b (1.1 g, 3.725 mmol) cyclobutanecarboxylate in EtOH (20 mL) containing a catalytic amount of Pd / C 10% (396 mg, 0.373 mmol) was hydrogenated under atmospheric pressure of H2 at room temperature for 4 h. The catalyst was removed by filtration over a short pad of Celite® and the filter cake was rinsed several times with MeOH. The combined filtrates were evaporated under reduced pressure to give 3- (3-amino-5-methoxyphenoxy) cyclobutanecarboxylate (1r, 3r) -ethyl 7c (920 mg). The compound was used as such in the next step. Synthesis of intermediate 7d:
[00181] [00181] A mixture of 2-bromo-2- (4-chlorophenyl) -1- (6- (trifluoromethoxy) indolin-1-yl) ethanone 1c (1.072 g, 2.467 mmol), 3- (3- amino-5 -methoxyphenoxy) (1r, 3r) -ethyl 7c cyclobutanecarboxylate (720 mg, 2.714 mmol) and diisopropylethylamine (850 µL, 4.934 mmol) in CH3CN (32 mL) was stirred at 60 ° C for 4 days. The mixture was diluted with water and extracted with EtOAc. The organic layer was washed with 1 N HCl and brine, dried over MgSO4, filtered and the solvent was evaporated in vacuo to give 3- (3 - ((1- (4-chlorophenyl) -2-oxo-2- (6 - (1r, 3r) -dyl (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) cyclobutanecarboxylate 7d (1.6 g), which was used as such in the next step. Synthesis of Compound 7 and chiral separation in Enantiomers 7A and 7B:
[00182] [00182] At 0 ° C, LiOH monohydrate (325 mg, 7.75 mmol) was added portion by portion to a solution of 3- (3 - ((1- (4-chlorophenyl) -2-oxo-2- ( 6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) (1r, 3r) -ethyl cyclobutanecarboxylate 7d (1.6 g, 2.585 mmol) in THF / water / MeOH (1/1 / 1) (30 mL). The reaction mixture was stirred at room temperature for 4 h. The mixture was diluted with water and 3 N HCl was added to acidify the solution. The mixture was extracted with EtOAc. The combined organic layers were dried over MgSO4, filtered and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (15-40 µm, 80 g, CH2Cl2 / MeOH 100/0 to 98/2). The product fractions were combined and evaporated under reduced pressure to give acid (1r, 3r) - 3- (3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethyl) indolin-1 -yl) ethyl) amino) -5-methoxyphenoxy) methyl) cyclobutene-carboxylic (Compound 7, 1.16 g). The two enantiomers were separated by chiral SFC (Stationary phase: Chiralcel® OD-H 5 µm 250 x 30 mm, mobile phase: 55% CO2, 45% MeOH) to give, after solidification from heptane / ether of diisopropyl, the Enantiomer eluted first 7A (358 mg) and the Enantiomer eluted second 7B (388 mg). Compound 7: 1
[00183] [00183] H NMR (500 MHz, DMSO-d6) į ppm 2.19 - 2.30 (m, 2 H) 2.57 (qd, J = 6.8, 3.6 Hz, 2 H) 2, 95 - 3.05 (m, 1 H) 3.08 - 3.27 (m, 2 H) 3.61 (s, 3 H) 4.03 (td, J = 10.4, 7.3 Hz, 1 H) 4.53 (td, J = 10.4, 6.3 Hz, 1 H) 4.68 (quin, J = 6.7 Hz, 1 H) 5.54 (d, J = 8.5 Hz, 1 H) 5.62 (s, 1 H) 5.85 (s, 1 H) 5.92 (s, 1 H) 6.53 (d, J = 8.5 Hz, 1 H) 7, 01 (dl, J = 7.9 Hz, 1 H) 7.33 (d, J = 8.2 Hz, 1 H) 7.45 (d, J = 8.2 Hz, 2 H) 7.55 ( d, J = 8.5 Hz, 2 H) 8.04 (s, 1 H) 12.34 (ls, 1 H)
[00184] [00184] LC / MS (LC-C method): Rt 2.95 min, MH + 591 7A enantiomer: 1
[00185] [00185] H NMR (500 MHz, DMSO-d6) į ppm 2.20 - 2.28 (m, 2 H) 2.53 - 2.60 (m, 2 H) 2.95 - 3.04 (m , 1 H) 3.07 - 3.26 (m, 2 H) 3.60 (s, 3 H) 4.03 (td, J = 10.4, 7.3 Hz, 1 H) 4.52 ( td, J = 10.2, 6.3 Hz, 1 H) 4.67 (t, J = 6.8 Hz, 1 H) 5.53 (d, J = 8.5 Hz, 1 H) 5, 61 (t, J = 2.0 Hz, 1 H) 5.84 (s, 1 H) 5.91 (s, 1 H) 6.52 (d, J = 8.5 Hz, 1 H) 7, 01 (dd, J = 8.2, 1.6 Hz, 1 H) 7.33 (d, J = 8.2 Hz, 1 H) 7.44 (d, J = 8.5 Hz, 2 H) 7.55 (d, J = 8.5 Hz, 2 H) 8.03 (s, 1 H) 12.24 - 12.40 (m, 1 H)
[00186] [00186] LC / MS (LC-C method): Rt 2.96 min, MH + 591
[00187] [00187] [Į] D20: -41.6 ° (c 0.298, DMF)
[00188] [00188] Chiral SFC (SFC-H method): Rt 1.25 min, MH + 591, chiral purity 100%. 7B Enantiomer: 1
[00189] [00189] H NMR (500 MHz, DMSO-d6) į ppm 2.19 - 2.28 (m, 2 H) 2.57 (qd, J = 6.8, 3.9 Hz, 2 H) 2, 95 - 3.04 (m, 1 H) 3.08 - 3.25 (m, 2 H) 3.60 (s, 3 H) 4.03 (td, J = 10.5, 7.1 Hz, 1 H) 4.53 (td, J = 10.4, 6.3 Hz, 1 H)
[00190] [00190] LC / MS (LC-C method): Rt 2.95 min, MH + 591
[00191] [00191] [Į] D20: + 43.7 ° (c 0.332, DMF)
[00192] [00192] Chiral SFC (SFC-H method): Rt 2.05 min, MH + 591, chiral purity 100%. Example 8: synthesis of (1s, 3s) -3- (3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) - 5-methoxyphenoxy) cyclobutene carboxylic (Compound 8) and separation in Enantiomers 8A and 8B Synthesis of intermediate 8a:
[00193] [00193] Under a flow of N2, to a solution of (1r, 3r) -ethyl [160351-88-2] 3-hydroxycyclobutanecarboxylate (1.86 g, 12.901 mmol) in CH2Cl2 (50 mL) was added pyridine ( 1.56 mL) and tosyl anhydride (4.63 g, 14.192 mmol). The mixture was stirred for 6 h at room temperature. The mixture was concentrated in vacuo, suspended in diethyl ether (200 ml) and washed with 0.5 M hydrochloric acid (2 × 60 ml), a saturated solution of sodium hydrogen carbonate (2 × 60 ml), water (60 mL) and brine (50 mL) and then dried over sodium sulfate, filtered and concentrated under reduced pressure to give 3- (tr-3-tosyloxy) cyclobutanecarboxylate (1r, 3r) -ethyl 8a (3.97 g). The compound was used as such in the next step. Synthesis of intermediate 8b:
[00194] [00194] 3- (Tosyloxy) cyclobutanecarboxylate (1r, 3r) -ethyl 8a (3.85 g, 12.904 mmol) was added dropwise to a mixture of 3-methoxy-5-nitrophenol [7145-49-5] (1.98 g, 11.73 mmol) and K2CO3 (1.95 g, 14.07 mmol) in DMF (20 mL). The mixture was stirred at 80 ° C for 16 h. The mixture was diluted with water and extracted with EtOAc. The organic layer was washed with water, dried over MgSO4, filtered and the solvent was evaporated under reduced pressure. Purification was performed by flash chromatography on silica gel (15-40 µm, 120 g, heptane / EtOAc 95/5 to 85/15). The pure fractions were combined and evaporated to dryness to give 3- (3-methoxy-5-nitrophenoxy) (1s, 3s) -ethyl 8b (2.04 g) cyclobutanecarboxylate (2.04 g). Synthesis of intermediate 8c:
[00195] [00195] A solution of 3- (3-methoxy-5-nitrophenoxy) (1s, 3s) -ethyl 8b (2.04 g, 6.908 mmol) cyclobutanecarboxylate in EtOH (50 mL) containing a catalytic amount of Pd / C 10% (735 mg, 0.691 mmol) was hydrogenated under atmospheric pressure of H2 at room temperature for 4 h. The catalyst was removed by filtration over a short pad of Celite® and the filter cake was rinsed several times with EtOH. The combined filtrates were evaporated under reduced pressure to give 3- (3-amino-5-methoxyphenoxy) cyclobutanecarboxylate (1s, 3s) -ethyl 8c (1.8 g). The compound was used as such in the next step. Synthesis of intermediate 8d:
[00196] [00196] A mixture of 2-bromo-2- (4-chlorophenyl) -1- (6- (trifluoromethoxy) indolin-1-yl) ethanone 1c (1.5 g, 3.462 mmol), 3- (3-amino - 5-methoxyphenoxy) (1s, 3s) -ethyl 8c cyclobutanecarboxylate (870 mg, 3.462 mmol) and diisopropylethylamine (1.19 mL, 6.924 mmol) in CH3CN (30 mL) was stirred at 80 ° C for 16 h . The mixture was diluted with water and extracted with EtOAc. The organic layer was washed with 1 N HCl and brine, dried over MgSO4, filtered and the solvent was evaporated in vacuo. The compound was crystallized from CH3CN / Et2O to give intermediate 8 (fraction 1, 820 mg). The filtrate was concentrated under reduced pressure to give another batch of crude intermediate 8 (fraction 2, 1 g).
[00197] [00197] The reaction was duplicated starting from 692 mg (1.592 mmol) of 1c (using the same reaction conditions as described above). The reaction product was crystallized from Et2O to give intermediate 8 (fraction 3, 400 mg). The filtrate was concentrated under reduced pressure to give another batch of intermediate 8 (fraction 4, 600 mg). Fractions 2 and 4 were combined and purified by flash chromatography on silica gel (15-40 µm, 80 g, heptane / EtOAc 90/10 at 70/30). The pure fractions were combined and evaporated to dryness to give fraction 5 (250 mg). Fractions 1, 3 and 5 were combined and dried to give 3- (3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethyl) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) (1s, 3s) -ethyl 8d cyclobutanecarboxylate (1.53 g). Synthesis of Compound 8 and chiral separation in Enantiomers 8A and 8B:
[00198] [00198] At 0 ° C, LiOH monohydrate (318 mg, 7.58 mmol) was added portion by portion to a solution of 3- (3 - ((1- (4-chlorophenyl) -2-oxo-2- ( 6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) (1s, 3s) -ethyl cyclobutanecarboxylate 8d (1.53 g, 2.529 mmol) in THF / water / MeOH (1/1 / 1) (30 mL). The reaction mixture was stirred at room temperature for 4 h. The mixture was diluted with water and 3 N HCl was added to acidify the solution. The mixture was extracted with EtOAc. The combined organic layers were dried over MgSO4, filtered and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (15-40 µm, 40 g, CH2Cl2 / MeOH 100/0 to 98.5 / 1.5). The pure fractions were combined and evaporated under reduced pressure to give (1s, 3s) -3- (3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethyl) indolin-1- acid il) ethyl) amino) -5-methoxyphenoxy) methyl) cyclobutanecarboxylic (Compound 8, 1.26 g). The two enantiomers were separated by chiral SFC (Stationary phase: Chiralcel® OD-H 5 µm 250 x 20 mm, mobile phase: 55% CO2, 45% EtOH) to give, after solidification from ether / ether of diisopropyl, the Enantiomer eluted first 8A (442 mg) and the Enantiomer eluted second 8B (433 mg). Compound 8: 1
[00199] [00199] H NMR (500 MHz, DMSO-d6) į ppm 2.03 - 2.13 (m, 2 H) 2.57 - 2.66 (m, 2 H) 2.66 - 2.76 (m , 1 H) 3.08 - 3.26 (m, 2 H) 3.61 (s, 3 H) 4.04 (td, J = 10.2, 7.3 Hz, 1 H) 4.44 - 4.58 (m, 2 H) 5.54 (d, J = 8.5 Hz, 1 H) 5.66 (s, 1 H) 5.87 (s, 1 H) 5.94 (s, 1 H) 6.49 (d, J = 8.5 Hz, 1 H) 7.02 (dl, J = 8.2 Hz, 1 H) 7.34 (d, J = 8.2 Hz, 1 H) 7.45 (d, J = 8.2 Hz, 2 H) 7.55 (d, J = 8.2 Hz, 2 H) 8.04 (s, 1 H) 12.27 (sl, 1 H)
[00200] [00200] LC / MS (LC-C method): Rt 2.90 min, MH + 591 8A enantiomer: 1
[00201] [00201] H NMR (500 MHz, DMSO-d6) į ppm 2.01 - 2.14 (m, 2 H) 2.55 - 2.72 (m, 3 H) 3.08 - 3.24 (m , 2 H) 3.61 (s, 3 H) 4.04 (td, J = 10.4, 7.3 Hz, 1 H) 4.41 - 4.57 (m, 2 H) 5.54 ( d, J = 8.8 Hz, 1 H) 5.66 (s, 1 H) 5.86 (s, 1 H) 5.93 (s, 1 H) 6.48 (dl, J = 8.8 Hz, 1 H) 7.01 (dl, J = 8.2 Hz, 1 H) 7.33 (d, J = 8.2 Hz, 1 H) 7.44 (d, J = 8.5 Hz, 2 H) 7.55 (d, J = 8.2 Hz, 2 H) 8.03 (s, 1 H)
[00202] [00202] LC / MS (LC-C method): Rt 2.90 min, MH + 591
[00203] [00203] [Į] D20: -47.1 ° (c 0.274, DMF)
[00204] [00204] Chiral SFC (SFC-I method): Rt 1.18 min, MH + 591, chiral purity 100%.
[00205] [00205] H NMR (500 MHz, DMSO-d6) į ppm 2.02 - 2.13 (m, 2 H) 2.55 - 2.70 (m, 3 H) 3.06 - 3.25 (m , 2 H) 3.61 (s, 3 H) 4.04 (td, J = 10.2, 7.3 Hz, 1 H) 4.39 - 4.57 (m, 2 H) 5.54 ( d, J = 8.8 Hz, 1 H) 5.66 (s, 1 H) 5.86 (s, 1 H) 5.93 (s, 1 H) 6.48 (dl, J = 8.8 Hz, 1 H) 7.01 (dl, J = 7.9 Hz, 1 H) 7.33 (d, J = 8.2 Hz, 1 H) 7.44 (d, J = 8.2 Hz, 2 H) 7.55 (d, J = 8.5 Hz, 2 H) 8.03 (s, 1 H)
[00206] [00206] LC / MS (LC-C method): Rt 2.91 min, MH + 591
[00207] [00207] [Į] D20: + 40.0 ° (c 0.25, DMF)
[00208] [00208] Chiral SFC (SFC-I method): Rt 2.16 min, MH + 591, chiral purity 100%. Example 9: synthesis of 3- (3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) methyl acid) bicycles [1.1.1] pentane-1-carboxylic (Compound 9) and chiral separation in Enantiomers 9A and 9B Synthesis of intermediate 9a:
[00209] [00209] 3-methoxy-5-nitrophenol [7145-49-5] (1.1 g, 6.4 mmol), di-tert-butyl azodicarboxylate (TBAD, 1.65 g, 7.04 mmol) and triphenylphosphine (2.35 g, 8.96 mmol) was dissolved in dry THF (25 mL) at room temperature under N2 atmosphere. A solution of methyl 3- (hydroxylmethyl) bicycle [1.1.1] pentane-1-carboxylate [180464-87-3]
[00210] [00210] A solution of methyl 3 - ((3-methoxy-5-nitrophenoxy)) bicyclo [1.1.1] pentane-1-carboxylate 9a (1.06 g, 3.44 mmol) in MeOH (150 mL ) containing a catalytic amount of 10% Pd / C (366 mg, 0.34 mmol) was hydrogenated under atmospheric pressure of H2 at room temperature for 10 min. The catalyst was removed by filtration over a short pad of Celite® under N2 atmosphere and the filter cake was rinsed several times with MeOH. The combined filtrates were evaporated to give methyl 3 - ((3-amino-5-methoxyphenoxy)) bicyclo [1.1.1] pentane-1-carboxylate 9b (961 mg) as a black oil which was used without further purification in next step. Synthesis of intermediate 9c:
[00211] [00211] A mixture of 2-bromo-2- (4-chlorophenyl) -1- (6- (trifluoromethoxy) indolin-1-yl) ethanone 1c (1.16 g, 2.67 mmol), 3 - (( Methyl 3-amino-5-methoxyphenoxy)) bicyclo [1.1.1] pentane-1-carboxylate 9b (961 mg, 3.47 mmol) and diisopropylethylamine (689 µL, 4.0 mmol) in CH3CN (50 ml) was stirred at 80 ° C overnight. The solvent was evaporated under reduced pressure and the residue was dissolved in CH2Cl2.
[00212] [00212] LiOH (92 mg, 3.84 mmol) was added to a solution of 3- ((3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1 -yl) ethyl) amino) - 5-methoxyphenoxy) methyl) bicyclo [1.1.1] methyl pentane-1-carboxylate 9c (1.21 g, 1.92 mmol) in a solvent mixture of MeOH (20 mL) , THF (40 ml) and water (20 ml). The reaction mixture was stirred at room temperature for 4 h. 1 N HCl (1 mL) was added and the organic volatiles were evaporated under reduced pressure. The residual aqueous mixture was diluted with water, acidified with 1 N HCl to pH 2 and extracted twice with Et2O. The combined organic layers were dried over MgSO4, filtered and evaporated under reduced pressure. The residue was dried under vacuum at 50 ° C to give 3 - (((3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoro-methoxy) indolin-1-yl) ethyl) acid) amino) -5-methoxyphenoxy) methyl) bicyclo [1.1.1] pentane-1-carboxylic (Compound 9, 1.06 g) as a pale yellow solid.
[00213] [00213] Compound 9 enantiomers (994 mg) were separated by preparative chiral SFC (Stationary phase: Chiralcel® Diacel OD 20 x 250 mm, mobile phase: CO2, EtOH + 0.4% iPrNH2). The product fractions were combined and evaporated under reduced pressure. The product eluted first was partitioned between EtOAc and water. 1 N HCl was added and the layers were separated. The aqueous layer was extracted again with EtOAc. The combined organic layers were dried over MgSO4, filtered, evaporated under reduced pressure and dried under vacuum at 50 ° C to give Enantiomer 9A (353 mg). The product eluted in second place was divided between EtOAc and water. 1 N HCl was added and the layers were separated. The aqueous layer was extracted again with EtOAc. The combined organic layers were dried over MgSO4, filtered, evaporated under reduced pressure and dried under vacuum at 50 ° C to give Enantiomer 9B (193 mg). Compound 9: 1
[00214] [00214] H NMR (360 MHz, DMSO-d6) į ppm 1.92 (s, 6 H) 3.06 - 3.27 (m, 2 H) 3.62 (s, 3 H) 3.87 ( s, 2 H) 4.06 (td, J = 10.3, 7.1 Hz, 1 H) 4.52 (td, J = 10.2, 6.6 Hz, 1 H) 5.58 (d , J = 8.8 Hz, 1 H) 5.75 (t, J = 2.0 Hz, 1 H) 5.92 - 5.97 (m, 2 H) 6.46 (d, J = 9, 1 Hz, 1 H) 7.01 (dd, J = 8.2, 1.6 Hz, 1 H) 7.34 (d, J = 8.1 Hz, 1 H) 7.41 - 7.47 ( m, 2 H) 7.51 - 7.58 (m, 2 H) 8.03 (ls, 1 H) 12.37 (ls, 1 H)
[00215] [00215] LC / MS (LC-A method): Rt 1.09 min, MH + 617 9A enantiomer: 1
[00216] [00216] H NMR (360 MHz, DMSO-d6) į ppm 1.92 (s, 6 H) 3.07 - 3.25 (m, 2 H) 3.61 (s, 3 H) 3.87 ( s, 2 H) 3.98 - 4.11 (m, 1 H) 4.51 (td, J = 10.2, 6.4 Hz, 1 H) 5.57 (d, J = 8.8 Hz , 1 H) 5.75 (t, J = 2.0 Hz, 1 H) 5.92 - 5.96 (m, 2 H) 6.46 (d, J = 9.1 Hz, 1 H) 7 .01 (dd, J = 8.1, 1.5 Hz, 1 H) 7.33 (d, J = 8.1 Hz, 1 H) 7.44 (d, J = 8.4 Hz, 2 H ) 7.50 - 7.59 (m, 2 H) 8.03 (ls, 1 H) 12.37 (ls, 1 H)
[00217] [00217] LC / MS (LC-B method): Rt 1.91 min, MH + 617
[00218] [00218] [Į] D20: -43.6 ° (c 0.5, DMF)
[00219] [00219] Chiral SFC (SFC-D method): Rt 5.26 min, MH + 617 chiral purity 98.6%. 9B Enantiomer:
[00220] [00220] H NMR (360 MHz, DMSO-d6) į ppm 1.93 (s, 6 H) 3.08 - 3.25 (m, 2 H) 3.62 (s, 3 H) 3.88 ( s, 2 H) 4.00 - 4.11 (m, 1 H) 4.52 (td, J = 10.2, 6.6 Hz, 1 H) 5.58 (d, J = 8.8 Hz , 1 H) 5.76 (t, J = 1.8 Hz, 1 H) 5.92 - 5.98 (m, 2 H) 6.47 (d, J = 8.8 Hz, 1 H) 7 , 01 (dd, J = 8.1, 1.5 Hz, 1 H) 7.33 (d, J = 8.4 Hz, 1 H) 7.44 (d, J = 8.4 Hz, 2 H ) 7.51 - 7.60 (m, 2 H) 8.04 (ls, 1 H) 12.38 (ls, 1 H)
[00221] [00221] LC / MS (LC-B method): Rt 1.91 min, MH + 617
[00222] [00222] [Į] D20: + 42.2 ° (c 0.41, DMF)
[00223] [00223] Chiral SFC (SFC-D method): Rt 6.47 min, MH + 617 chiral purity 99.5%. Example 10: synthesis of (1s, 3s) -3 - ((3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoro-methoxy) indolin-1-yl) ethyl) acid) amino) -5- methoxyphenoxy) methyl) cyclobutenocarboxylic (Compound 10) and chiral separation in Enantiomers 10A and 10B Synthesis of intermediate 10a:
[00224] [00224] Under a flow of N2, to a solution of methyl 3- (hydroxymethyl) cyclobutanecarboxylate [89941-55-9] (1.4 g, 9.71 mmol) in CH2Cl2 (20 mL) was added pyridine (1 , 17 mL) and tosyl anhydride (3.49 g, 10.682 mmol). The mixture was stirred for 16 h at room temperature. The mixture was concentrated in vacuo, suspended in diethyl ether (200 ml) and washed with hydrochloric acid at
[00225] [00225] 3 - ((Tosyloxy) methyl) cyclobutanecarboxylate 10a (2.15 g, 7.206 mmol) was added dropwise to a mixture of 3-methoxy-5-nitrophenol [7145-49-5] (1.22 g , 7.206 mmol) and K2CO3 (1.5 g, 10.809 mmol) in DMF (14 mL). The mixture was stirred at 60 ° C for 16 h. The mixture was diluted with water and extracted with EtOAc. The organic layer was washed with water, dried over MgSO4, filtered and the solvent was evaporated under reduced pressure. Purification by flash chromatography on silica gel (15-40 µm, 80 g, heptane / EtOAc 85/15 to 80/20) was performed. The pure fractions were combined and evaporated to dryness. The stereoisomers were separated by chiral SFC (Stationary phase: Chiralpak® AD-H 5 µm 250 x 30 mm, mobile phase: 55% CO2, 45% MeOH) to give 3 - ((3-methoxy-5-nitrophenoxy) ) (1s, 3s) -methyl 10b (541 mg) and 3 - ((3-methoxy-5-nitrophenoxy) methyl) (1r, 3r) - methyl 10c (428 mg) cyclobutanecarboxylate. Synthesis of intermediate 10d:
[00226] [00226] A solution of 3 - ((3-methoxy-5-nitrophenoxy) methyl) (1s, 3s) -methyl 10b (530 mg, 1,795 mmol) cyclobutanecarboxylate in MeOH (10 mL) containing a catalytic amount of Pd / 10% C (191 mg, 0.179 mmol) was hydrogenated under atmospheric pressure of H2 at room temperature for 4 h. The catalyst was removed by filtration over a short pad of Celite® and the filter cake was rinsed several times with MeOH. The combined filtrates were evaporated under reduced pressure to give 3 - ((3-
[00227] [00227] A mixture of 2-bromo-2- (4-chlorophenyl) -1- (6- (trifluoromethoxy) indolin-1-yl) ethanone 1c (580 mg, 1.334 mmol), 3 - ((3-amino- 5-methoxyphenoxy) methyl) cyclobutanecarboxylate (1s, 3s) -methyl 10d (460 mg, 1.734 mmol) and diisopropylethylamine (460 µL, 2.667 mmol) in CH3CN (10 mL) was stirred at 60 ° C for 48 h. The mixture was concentrated under reduced pressure and absorbed with EtOAc. The organic layer was washed with 1 N HCl and water, dried over MgSO4, filtered and the solvent was evaporated in vacuo to give, after crystallization from Et2O / diisopropyl ether, 3 - (((3 - ((1 - (1s, 3s) -methyl (1s, 3s) -methyl (6s-3s) -methyl 10e (625-(4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) mg). Synthesis of Compound 10 and chiral separation in Enantiomers 10A and 10B:
[00228] [00228] At 0 ° C, LiOH monohydrate (127 mg, 3.029 mmol) was added portion by portion to a solution of 3- (3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) (1s, 3s) -methyl 10e (625 mg, 1.01 mmol) cyclobutanecarboxylate in THF / water / MeOH (1/1/1) (15 mL). The reaction mixture was stirred at room temperature for 48 h. The mixture was cooled to 0 ° C and was diluted with water. 3 N HCl was added to acidify the solution and the mixture was extracted with EtOAc. The combined organic layers were dried over MgSO4, filtered and concentrated under reduced pressure. The residue was crystallized from Et2O / diisopropyl ether to give (1s, 3s) -3 - ((3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy ) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) methyl) cyclobutene carboxylic (Compound 10, 440 mg). The two enantiomers were separated by chiral SFC (Stationary phase: Whelk® O1 (S, S)
[00229] [00229] H NMR (500 MHz, DMSO-d6) į ppm 1.87 - 1.99 (m, 2 H) 2.18 - 2.30 (m, 2 H) 2.53 - 2.61 (m , 1 H) 2.97 (quin, J = 8.9 Hz, 1 H) 3.08 - 3.27 (m, 2 H) 3.62 (s, 3 H) 3.78 (dl, J = 6.3 Hz, 2 H) 4.05 (td, J = 10.4, 7.2 Hz, 1 H) 4.52 (td, J = 10.3, 6.5 Hz, 1 H) 5, 57 (d, J = 8.8 Hz, 1 H) 5.72 - 5.78 (m, 1 H) 5.94 (s, 2 H) 6.45 (dl, J = 8.8 Hz, 1 H) 6.95 - 7.06 (m, 1 H) 7.34 (d, J = 8.2 Hz, 1 H) 7.44 (d, J = 8.2 Hz, 2 H) 7.55 (d, J = 8.5 Hz, 2 H) 8.04 (s, 1 H) 12.08 (sl, 1 H)
[00230] [00230] LC / MS (LC-D method): Rt 2.94 min, MH + 605
[00231] [00231] PF = 128 ° C 10A enantiomer: 1
[00232] [00232] H NMR (500 MHz, DMSO-d6) į ppm 1.84 - 1.97 (m, 2 H) 2.15 - 2.30 (m, 2 H) 2.53 - 2.61 (m , 1 H) 2.94 (quin, J = 8.9 Hz, 1 H) 3.07 - 3.25 (m, 2 H) 3.61 (s, 3 H) 3.77 (dl, J = 6.3 Hz, 2 H) 4.05 (td, J = 10.4, 6.9 Hz, 1 H) 4.52 (td, J = 10.3, 6.1 Hz, 1 H) 5, 57 (d, J = 8.8 Hz, 1 H) 5.74 (t, J = 1.9 Hz, 1 H) 5.93 (s, 2 H) 6.44 (d, J = 8.8 Hz, 1 H) 7.01 (dd, J = 8.0, 1.4 Hz, 1 H) 7.33 (d, J = 8.2 Hz, 1 H) 7.44 (d, J = 8 , 5 Hz, 2 H) 7.55 (d, J = 8.2 Hz, 2 H) 8.03 (s, 1 H)
[00233] [00233] LC / MS (LC-C method): Rt 3.08 min, MH + 605
[00234] [00234] [Į] D20: -44.0 (c 0.314, DMF)
[00235] [00235] Chiral SFC (SFC-J method): Rt 1.63 min, MH + 605, chiral purity 100%. 10B Enantiomer: 1
[00236] [00236] H NMR (500 MHz, DMSO-d6) į ppm 1.85 - 1.97 (m, 2 H) 2.17 - 2.28 (m, 2 H) 2.53 - 2.58 (m , 1 H) 2.94 (quint, J = 8.8 Hz, 1 H) 3.07 - 3.24 (m, 2 H) 3.61 (s, 3 H) 3.72 - 3.82 ( m, 2 H) 4.05 (td, J = 10.4, 7.3 Hz,
[00237] [00237] LC / MS (LC-C method): Rt 3.08 min, MH + 605
[00238] [00238] [Į] D20: + 45.5 ° (c 0.308, DMF)
[00239] [00239] Chiral SFC (SFC-J method): Rt 2.14 min, MH + 605, chiral purity 99.57%. Example 11: synthesis of (1r, 3r) -3 - ((3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino acid) -5-methoxyphenoxy) methyl) cyclobutene carboxylic (Compound 11) and chiral separation in Enantiomers 11A and 11B Synthesis of intermediate 11a:
[00240] [00240] A solution of 3 - ((3-methoxy-5-nitrophenoxy) methyl) cyclobutene-carboxylate (1r, 3r) -methyl 10c (410 mg, 1.388 mmol) in MeOH (10 mL) containing a catalytic amount of 10% Pd / C (148 mg, 0.139 mmol) was hydrogenated under atmospheric pressure of H2 at room temperature for 4 h. The catalyst was removed by filtration over a short pad of Celite® and the filter cake was rinsed several times with MeOH. The combined filtrates were evaporated under reduced pressure to give (1r, 3r) 3 - ((3-amino-5-methoxyphenoxy) methyl) cyclobutanecarboxylate (370 mg).
[00241] [00241] A mixture of 2-bromo-2- (4-chlorophenyl) -1- (6- (trifluoromethoxy) indolin-1-yl) ethanone 1c (460 mg, 1.058 mmol), 3 - ((3- amino- 5-methoxyphenoxy) methyl) cyclobutanecarboxylate (1r, 3r) -methyl 11a (365 mg, 1.376 mmol) and diisopropylethylamine (365 µL, 2.117 mmol) in CH3CN (8 mL) was stirred at 60 ° C for 48 h. The mixture was concentrated under reduced pressure and absorbed with EtOAc. The organic layer was washed with 1 N HCl and water, dried over MgSO4, filtered and the solvent was evaporated in vacuo to give, after crystallization from Et2O / diisopropyl ether, 3 - (((3 - ((1 - (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) methyl) (1r, 3r) -methyl 11b (515 mg) cyclobutanecarboxylate . Synthesis of Compound 11 and chiral separation in Enantiomers 11A and 11B:
[00242] [00242] At 0ºC, LiOH monohydrate (105 mg, 2.496 mmol) was added portion by portion to a solution of 3- (3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) (1r, 3r) -methyl 11b (515 mg, 0.832 mmol) cyclobutanecarboxylate in THF / water / MeOH (1/1/1) (15 mL). The reaction mixture was stirred at room temperature for 48 h. The mixture was cooled to 0 ° C and was diluted with water. 3 N HCl was added to acidify the solution and the mixture was extracted with EtOAc. The combined organic layers were dried over MgSO4, filtered and concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel (15-40 µm, 24 g, CH2Cl2 / CH3OH 100/0 to 98/2). The pure fractions were combined and evaporated to dryness to give (1r, 3r) -3 - ((3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethyl) indolin-1 acid - yl) ethyl) amino) -5-methoxyphenoxy) methyl) cyclobutanecarboxylic (Compound 11, 460 mg). The two enantiomers were separated by chiral SFC
[00243] [00243] H NMR (500 MHz, DMSO-d6) į ppm 1.92 - 2.04 (m, 2 H) 2.21 - 2.31 (m, 2 H) 2.57 - 2.62 (m , 1 H) 3.04 - 3.26 (m, 3 H) 3.63 (s, 3 H) 3.88 (dl, J = 6.9 Hz, 2 H) 4.00 - 4.11 ( m, 1 H) 4.47 - 4.58 (m, 1 H) 5.57 (d, J = 8.8 Hz, 1 H) 5.78 (s, 1 H) 5.96 (dl, J = 9.7 Hz, 2 H) 6.45 (dl, J = 8.8 Hz, 1 H) 7.02 (dl, J = 8.2 Hz, 1 H) 7.34 (d, J = 8 , 2 Hz, 1 H) 7.45 (d, J = 8.2 Hz, 2 H) 7.55 (d, J = 8.5 Hz, 2 H) 8.04 (s, 1 H) 12, 12 (ls, 1 H)
[00244] [00244] LC / MS (LC-D method): Rt 2.95 min, MH + 605 11A enantiomer: 1
[00245] [00245] H NMR (500 MHz, DMSO-d6) į ppm 1.94 - 2.03 (m, 2 H) 2.21 - 2.31 (m, 2 H) 2.55 - 2.62 (m , 1 H) 3.04 - 3.25 (m, 3 H) 3.62 (s, 3 H) 3.87 (d, J = 7.3 Hz, 2 H) 4.04 (td, J = 10.4, 6.9 Hz, 1 H) 4.52 (td, J = 10.2, 6.3 Hz, 1 H) 5.57 (d, J = 8.8 Hz, 1 H) 5, 78 (t, J = 1.9 Hz, 1 H) 5.94 (s, 1 H) 5.96 (s, 1 H) 6.45 (d, J = 8.5 Hz, 1 H) 7, 01 (dd, J = 8.2, 1.6 Hz, 1 H) 7.33 (d, J = 8.2 Hz, 1 H) 7.44 (d, J = 8.5 Hz, 2 H) 7.55 (d, J = 8.5 Hz, 2 H) 8.03 (s, 1 H) 12.12 (ls, 1 H)
[00246] [00246] LC / MS (LC-C method): Rt 3.08 min, MH + 605
[00247] [00247] [Į] D20: -43.3 ° (c 0.319, DMF)
[00248] [00248] Chiral SFC (SFC-J method): Rt 1.73 min, MH + 605, 100% chiral purity. 11B Enantiomer: 1
[00249] [00249] H NMR (500 MHz, DMSO-d6) į ppm 1.92 - 2.02 (m, 2 H) 2.21 - 2.32 (m, 2 H) 2.55 - 2.62 (m , 1 H) 3.04 - 3.25 (m, 3 H) 3.62 (s, 3 H) 3.87 (d, J = 6.9 Hz, 2 H) 4.05 (td, J = 10.3, 7.1 Hz, 1 H) 4.52 (td, J = 10.3, 6.5 Hz, 1 H) 5.57 (d, J = 8.8 Hz, 1 H) 5, 78 (t, J = 1.9 Hz, 1 H) 5.94 (s, 1 H) 5.96 (s, 1 H) 6.45 (dl, J = 8.8 Hz, 1 H) 7, 01 (dl, J = 8.2 Hz, 1 H) 7.33 (d,
[00250] [00250] LC / MS (LC-C method): Rt 3.08 min, MH + 605
[00251] [00251] [Į] D20: + 45.5 ° (c 0.323, DMF)
[00252] [00252] Chiral SFC (SFC-J method): Rt 2.36 min, MH + 605, chiral purity 99.61%. Example 12: synthesis of 3- (3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) propanoic acid ( Compound 12) and chiral separation in Enantiomers 12A and 12B Synthesis of intermediate 12a:
[00253] [00253] 1,4-Diazabicyclo [2.2.2] octane (66 mg, 0.591 mmol) was added to a solution of 3-methoxy-5-nitrophenol [7145-49-5] (2.0 g, 11.825 mmol) , ethylacetylenecarboxylate (1.2 ml, 11.825 mmol) in CH2Cl2 (20 ml). The mixture was stirred at room temperature for 16 h. The mixture was poured into water and the layers were decanted. The organic layer was washed with water, brine, dried over MgSO4, filtered and the solvent was evaporated under reduced pressure to give ethyl 3- (3-methoxy-5-nitrophenoxy) acrylate 12a (3.22 g, E / Z mixture) ). Synthesis of intermediate 12b:
[00254] [00254] A mixture of 3- (3-methoxy-5-nitrophenoxy) ethyl acrylate 12a (3.2 g, 11.97 mmol) and Pd / C (10%) (2.5 g, 2.395 mmol) in CH3OH (100 ml) was hydrogenated under a pressure of 3 bar for 3 h. The catalyst was removed by filtration through a pad of Celite®. The filter cake was rinsed with CH3OH and the combined filtrates were concentrated under reduced pressure. Purification was carried out by flash chromatography on silica gel (15-40 µm, 80 g, heptane / EtOAc 75/25). The pure fractions were combined and evaporated to dryness to give ethyl 3- (3-amino-5-methoxyphenoxy) propanoate 12b (1.8 g). Synthesis of intermediate 12c:
[00255] [00255] A mixture of 2-bromo-2- (4-chlorophenyl) -1- (6- (trifluoromethoxy) indolin-1-yl) ethanone 1c (1.0 g, 2.301 mmol), 3- (3-amino - Ethyl 5-methoxyphenoxy) -propanoate 12b (716 mg, 2.991 mmol) and diisopropylethylamine (793 µL, 4.602 mmol) in CH3CN (29 mL) was stirred at 60 ° C for 16 h. The mixture was concentrated under reduced pressure and absorbed with EtOAc. The organic layer was washed with 1 N HCl and water, dried over MgSO4, filtered and the solvent was evaporated in vacuo. Purification was carried out by flash chromatography on silica gel (15-40 µm, 80 g, heptane / EtOAc 75/25). The pure fractions were combined and evaporated to dryness to give 3- (3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino acid) -5-methoxyphenoxy) propanoic 12c (660 mg). Synthesis of Compound 12 and chiral separation in Enantiomers 12A and 12B:
[00256] [00256] At 0 ° C, LiOH monohydrate (79 mg, 1.889 mmol) was added portion by portion to a solution of 3- (3 - ((1- (4-chlorophenyl) -2-oxo-2- (6 - (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) propanoic 12c (560 mg, 0.944 mmol) in THF / water / MeOH (1/1/1) (15 mL). The reaction mixture was stirred at room temperature for 2 h. The mixture was cooled to 0 ° C and was diluted with water. 3 N HCl was added to acidify the solution. The mixture was extracted with EtOAc. The combined organic layers were dried over MgSO4, filtered and concentrated under reduced pressure. The residue was purified through a reverse phase (Stationary phase: YMC-DispoPack AT ODS-25: 120 g, mobile phase: 75% NH4HCO3 gradient, 0.2%, 25% CH3CN to 35% NH4HCO3, 0, 2%, 65% CH3CN). The pure fractions were combined and evaporated to dryness to give 3- (3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino acid) -5-methoxyphenoxy) propanoic (Compound 12, 126 mg). The two enantiomers were separated into a 70 mg batch using chiral SFC (Stationary phase: Chiralcel® OD-H 5 µm 250 x 20 mm, mobile phase: 65% CO2, 35% iPrOH (+ 0.3 iPrNH2 %) to give, after lyophilization in a mixture of CH3CN (2 ml) / water (8 ml), the Enantiomer eluted first 12A (30 mg) and the Enantiomer eluted second 12B (35 mg). 1
[00257] [00257] H NMR (400 MHz, DMSO-d6) į ppm 2.56 (t, J = 6.1 Hz, 2 H) 3.04 - 3.26 (m, 2 H) 3.61 (s, 3 H) 3.95 - 4.11 (m, 3 H) 4.43 - 4.61 (m, 1 H) 5.56 (d, J = 8.6 Hz, 1 H) 5.74 (s , 1 H) 5.94 (dl, J = 7.1 Hz, 2 H) 6.46 (d, J = 9.1 Hz, 1 H) 7.00 (dl, J = 8.1 Hz, 1 H) 7.33 (d, J = 8.1 Hz, 1 H) 7.43 (d, J = 8.1 Hz, 2 H) 7.55 (d, J = 8.6 Hz, 2 H) 8.03 (s, 1 H)
[00258] [00258] LC / MS (LC-C method): Rt 2.84 min, MH + 565 12A Enantiomer: 1
[00259] [00259] H NMR (500 MHz, DMSO-d6) į ppm 2.62 (tl, J = 6.0 Hz, 2 H) 3.09 - 3.24 (m, 2 H) 3.62 (s, 3 H) 4.00 - 4.10 (m, 3 H) 4.47 - 4.57 (m, 1 H) 5.58 (d, J = 8.8 Hz, 1 H) 5.75 (s , 1 H) 5.95 (dl, J = 6.6 Hz, 2 H) 6.49 (dl, J = 8.8 Hz, 1 H) 7.02 (dl, J = 8.2 Hz, 1 H) 7.34 (d, J = 8.2 Hz, 1 H) 7.44 (d, J = 8.5 Hz, 2 H) 7.55 (d, J = 8.5 Hz, 2 H) 8.04 (s, 1 H) 12.04 - 12.63 (m, 1 H)
[00260] [00260] LC / MS (LC-C method): Rt 2.83 min, MH + 565
[00261] [00261] [Į] D20: -47.3 ° (c 0.275, DMF)
[00262] [00262] Chiral SFC (SFC-K method): Rt 2.50 min, MH + 565, chiral purity 100%. 12B Enantiomer: 1
[00263] [00263] H NMR (500 MHz, DMSO-d6) į ppm 2.58 - 2.67 (m, 2 H) 3.08 - 3.25 (m, 2 H) 3.62 (s, 3 H) 3.99 - 4.12 (m, 3 H) 4.52 (td, J = 10.3, 6.1 Hz, 1 H) 5.58 (s, 1 H) 5.75 (s, 1 H ) 5.95 (dl, J = 6.6 Hz, 2 H) 6.50 (ls, 1 H) 7.02 (dl, J = 7.9 Hz, 1 H) 7.34 (d, J = 8.2 Hz, 1 H) 7.44 (d, J = 8.5 Hz, 2 H) 7.55 (d, J = 8.5 Hz, 2 H) 8.04 (s, 1 H) 12 , 35 (ls, 1 H)
[00264] [00264] LC / MS (LC-C method): Rt 2.83 min, MH + 565
[00265] [00265] [Į] D20: + 41.8 ° (c 0.297, DMF)
[00266] [00266] Chiral SFC (SFC-K method): Rt 4.34 min, MH + 565, chiral purity 99.1%. Example 13: synthesis of 5- (3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) pentanoic acid ( Compound 13) and chiral separation in Enantiomers 13A and 13B Synthesis of intermediate 13a:
[00267] [00267] To a stirred solution of methyl 5-bromo-valerate [CAS 5454-83-1] (1.06 mL, 7.19 mmol) in DMF (25 mL) were added 3-amino-5-methoxyphenol [ CAS 162155-27-3] (1.0 g, 7.19 mmol) and Cs2CO3 (4.68 g, 14.4 mmol). The reaction was stirred at 60 ° C for 18 h, and allowed to reach room temperature. The mixture was poured into H2O (125 ml). The product was extracted (2 x) with CH2Cl2. The combined organic layers were dried over MgSO4, filtered, and the solvent was evaporated under reduced pressure. The residue was purified by flash chromatography on silica gel (25 g) using a gradient of heptane / EtOAc 100/0 at 50/50. The product fractions were combined, evaporated under reduced pressure and coevaporated with CH3CN. The product was dried under vacuum at 45 ° C to provide methyl 5- (3-amino-5-methoxyphenoxy) pentanoate 13a (200 mg). Synthesis of intermediate 13b:
[00268] [00268] A mixture of 2-bromo-2- (4-chlorophenyl) -1- (6- (trifluoromethoxy) indolin-1-yl) ethanone 1c (340 mg, 0.78 mmol), 5- (3-amino -5-methoxyphenoxy) methyl pentanoate 13a (198 mg, 0.78 mmol) and diisopropylethylamine (270 µL, 1.56 mmol) in CH3CN (30 mL) was stirred at 60 ° C for 18 h. The mixture was allowed to reach room temperature and was poured into water (150 ml). The product was extracted (2x) with Et2O. The combined organic layers were washed with brine, dried over MgSO4, filtered and evaporated under reduced pressure. The residue was purified by flash chromatography on silica gel (40 g) using a gradient of heptane / EtOAc / EtOH 100/0/0 to 40/45/15. The product fractions were combined and evaporated under reduced pressure and coevaporated with dioxane to provide 5- (3- ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) methyl pentanoate 13b (475 mg). Synthesis of Compound 13 and separation in Enantiomers 13A and 13B:
[00269] [00269] 1 M NaOH in water (1.96 mL, 1.96 mmol) was added to a stirring solution of 5- (3 - ((1- (4-chlorophenyl) -2-oxo-2- ( 6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) methyl pentanoate 13b (475 mg, 0.78 mmol) in dioxane (5 mL). The reaction mixture was stirred at room temperature for 4.5 h. The reaction mixture was diluted with water (20 ml), 1 N HCl (2.1 ml). After stirring for 10 min, the product was extracted with 2-Me-THF. The organic layer was separated, dried over MgSO4, filtered and evaporated under reduced pressure. The residue was purified by flash chromatography on silica gel (40 g) with a gradient of heptane / EtOAc / EtOH / HOAc 100/0/0/0 to 40/45 / 14.7 / 0.3. The product fractions were combined and evaporated under reduced pressure to a residual volume of ~ 7.5 ml, allowing precipitation of the reaction product. The solids were filtered off, washed (3x) with 1/3 EtOAc / heptane and dried under vacuum at 45 ° C to provide 5- (3 - ((1- (4-chlorophenyl) -2-oxo-2- ( 6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) pentanoic (Compound 13, 139 mg) as a racemic mixture.
[00270] [00270] Compound 13 enantiomers (112 mg) were separated by preparative chiral SFC (Stationary phase: Chiralpak® Diacel OD 20 x 250 mm, mobile phase: CO2, EtOH + 0.4% iPrNH2). The fractions containing the first eluted product were combined, evaporated under reduced pressure and coevaporated with CH3CN. The residue was lyophilized from a solvent mixture of CH3CN (1.5 ml) and water (1 ml) to provide Enantiomer 13A (39 mg). The fractions containing the second eluted product were combined, evaporated under reduced pressure and evaporated with CH3CN. The residue was lyophilized from CH3CN (1.75 ml) and water (1.25 ml) to provide Enantiomer 13B (33 mg). Compound 13 1
[00271] [00271] H NMR (400 MHz, DMSO-d6) į ppm 1.55 - 1.69 (m, 4 H) 2.25 (t, J = 7.0 Hz, 2 H) 3.07 - 3, 26 (m, 2 H) 3.62 (s, 3 H) 3.83 (t, J = 5.9 Hz, 2 H) 4.05 (td, J = 10.4, 7.2 Hz, 1 H) 4.52 (td, J = 10.3, 6.4 Hz, 1 H) 5.56 (d, J = 8.8 Hz, 1 H) 5.75 (t, J = 2.0 Hz , 1 H) 5.91 - 5.97 (m, 2 H) 6.42 (d, J = 8.8 Hz, 1 H) 7.01 (dd, J = 8.3, 1.7 Hz, 1 H) 7.33 (d, J = 8.1 Hz, 1 H) 7.38 - 7.48 (m, 2 H) 7.55 (d, J = 8.6 Hz, 2 H) 8, 03 (sl, 1 H) 12.01 (s, 1 H)
[00272] [00272] LC / MS (LC-A method): Rt 1.13 min, MH + 593 13A enantiomer: 1
[00273] [00273] H NMR (400 MHz, DMSO-d6) į ppm 1.55 - 1.69 (m, 4 H)
[00274] [00274] LC / MS (LC-B method): Rt 2.08 min, MH + 593
[00275] [00275] [Į] D20: -48.6 ° (c 0.43, DMF)
[00276] [00276] Chiral SFC (SFC-D method): Rt 5.27 min, MH + 593 100% chiral purity. 13B Enantiomer: 1
[00277] [00277] H NMR (400 MHz, DMSO-d6) į ppm 1.55 - 1.69 (m, 4 H) 2.24 (t, J = 7.2 Hz, 2 H) 3.04 - 3, 25 (m, 2 H) 3.62 (s, 3 H) 3.83 (t, J = 5.9 Hz, 2 H) 4.05 (td, J = 10.4, 7.2 Hz, 1 H) 4.52 (td, J = 10.3, 6.2 Hz, 1 H) 5.56 (d, J = 8.8 Hz, 1 H) 5.75 (t, J = 2.0 Hz , 1 H) 5.92 - 5.96 (m, 2 H) 6.42 (d, J = 8.8 Hz, 1 H) 7.01 (dd, J = 8.1, 1.5 Hz, 1 H) 7.33 (d, J = 8.1 Hz, 1 H) 7.42 - 7.46 (m, 2 H) 7.55 (d, J = 8.6 Hz, 2 H) 8, 03 (sl, 1 H) 12.00 (sl, 1 H)
[00278] [00278] LC / MS (LC-B method): Rt 2.08 min, MH + 593
[00279] [00279] [Į] D20: + 48.3 ° (c 0.42, DMF)
[00280] [00280] Chiral SFC (SFC-D method): Rt 6.94 min, MH + 593 100% chiral purity. Example 14: synthesis of 5- (3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoro-methoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenyl acid) pentanoic (Compound 14) and chiral separation in Enantiomers 14A and 14B
[00281] [00281] In a sealed tube under an N2 atmosphere, PdCl2 (PPh3) 2 (1.5 g, 2.2 mmol) was added to a degassed solution of 1-bromo-3-methoxy-5-nitrobenzene [CAS 16618- 67-0] (5.0 g, 22 mmol), methyl acrylate (6.0 mL, 67 mmol) in CH3CN (45 mL) and triethylamine (12 mL). The reaction was stirred at 70 ° C for 18 h. The mixture was concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel (30 µm, 120 g, heptane / EtOAc gradient from 90/10 to 75/25) to give 3- (3) -methoxy-5-nitrophenyl) acrylate -methyl 14a (2.0 g). Synthesis of intermediate 14b:
[00282] [00282] Under N2 at 0 ° C, diisobutylaluminum hydride (1 M in CH2Cl2) (20 mL, 20 mmol) was added dropwise to a solution of 3- (3-methoxy-5-nitrophenyl) acrylate ( E) -methyl 14a (2.4 g, 10.12 mmol) in CH2Cl2 (65 mL). The reaction was slowly warmed to room temperature and stirred for 2 h. The mixture was quenched with HCl (3 N) and extracted with CH2Cl2. The organic layer was dried over MgSO4, filtered and concentrated under reduced pressure to give (E) -3- (3-methoxy-5-nitrophenyl) prop-2-en-1-ol 14b (2.1 g). The compound was used as such in the next step.
[00283] [00283] Under N2 at 0 ° C, Dess-Martin periodinane (24 mL, 11.04 mmol) was slowly added to a solution of (E) -3- (3-methoxy-5-nitrophenyl) prop-2- en-1-ol 14b (2.1 g, 10.04 mmol) in CH2Cl2 (64 mL) and the mixture was stirred at room temperature for 72 h. Water was added and the mixture was filtered. The filtrate was extracted with CH2Cl2, dried over MgSO4 and evaporated to dryness to give 14c (2.5 g) (E) -3- (3-methoxy-5-nitrophenyl) acrylaldehyde. The compound was used as such in the next step. Synthesis of intermediate 14d:
[00284] [00284] Under N2, (carbetoxymethylene) triphenylphosphorane (5.0 g, 14.48 mmol) was added in one portion to a mixture of (E) -3- (3-methoxy-5-nitrophenyl) acrylaldehyde 14c (2, 5 g, 9.65 mmol, 80% pure) in CH2Cl2 (62 mL) and stirred at room temperature for 16 h. The mixture was concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel (30 µm, 80 g, heptane / EtOAc gradient from 85/15 to 60/30) to give 5- (3-methoxy-5-nitrophenyl) penta-2,4 (2E, 4E) -ethyl dienoate 14d (2.1 g). The compound was used as such in the next step. Synthesis of intermediate 14e:
[00285] [00285] A mixture of 5- (3E, 4E) -ethyl 5- (3-methoxy-5-nitrophenyl) penta-2,4-dienoate (1.9 g, 6.85 mmol) in EtOH (40 mL) and EtOAc (6.7 ml) was hydrogenated under an atmospheric pressure of H2 for 18 h with Pd / C (10%) (0.73 g, 0.69 mmol) as a catalyst. The catalyst was removed by filtration through a pad of Celite®. Celite® was washed with EtOAc and the combined filtrates were concentrated in vacuo. The residue was hydrogenated again in EtOH (40 ml) and EtOAc (6.7 ml) under an atmospheric pressure of H2 for 72 h with Pd / C (10%) (0.73 g, 0.69 mmol) as a catalyst . The catalyst was removed by filtration through a pad of Celite®. Celite® was washed with EtOAc and the combined filtrates were concentrated in vacuo to give ethyl 5- (3-amino-5-methoxyphenyl) pentanoate (1.4 g). The compound was used as such in the next step. Synthesis of intermediate 14f:
[00286] [00286] A mixture of 2-bromo-2- (4-chlorophenyl) -1- (6- (trifluoromethoxy) indolin-1-yl) ethanone 1c (1.6 g, 3.71 mmol), 5- (3 ethyl 14-amino-5-methoxyphenyl) pentanoate (1.4 g, 5.57 mmol) and diisopropylethylamine (1.3 mL, 7.43 mmol) in CH3CN (19 mL) was stirred at 80 ° C for 18 h. The mixture was absorbed with EtOA and washed with 0.5 N HCl (twice) and water. The organic layer was dried over MgSO4, filtered and the solvent was evaporated in vacuo. Purification was performed by flash chromatography on silica gel (30 µm, 40 g, heptane / EtOAc from 85/15 to 75/25) to give 5- (3 - ((1- (4-chlorophenyl) -2-oxo- 2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenyl) pentanoate 14f (1.8 g). The compound was used as such in the next step. Synthesis of Compound 14 and chiral separation in Enantiomers 14A and 14B:
[00287] [00287] A solution of LiOH monohydrate (0.62 g, 15 mmol) in water (16 mL) was added dropwise to a solution of 5- (3 - ((1- (4-chlorophenyl) -2 -oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenyl) 14f ethyl pentanoate (1.8 g, 2.98 mmol) in THF (36 mL). The mixture was stirred at room temperature for 18h. LiOH monohydrate (0.62 g, 15 mmol) was added again and the mixture was stirred at room temperature for 24 h. The solution was acidified with HCl (3 N) and extracted with EtOAc. The organic layer was dried over MgSO4, filtered and concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel (30 µm, 40 g, CH2Cl2 / MeOH 100/0 to 98/2) to give 5- (3 - ((1- (4-chlorophenyl) -2-oxo acid - 2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenyl) pentanoic (compound 14, 920 mg). The enantiomers were separated by
[00288] [00288] H NMR (500 MHz, DMSO-d6) į ppm 1.47 (ls, 4 H) 2.15 - 2.22 (m, 2 H) 2.34 - 2.43 (m, 2 H) 3.07 - 3.26 (m, 2 H) 3.62 (s, 3 H) 3.92 - 4.14 (m, 1 H) 4.40 - 4.64 (m, 1 H) 5, 56 (dl, J = 8.83 Hz, 1 H) 6.00 (s, 1 H) 6.13 (s, 1 H) 6.20 (s, 1 H) 6.37 (dl, J = 8 , 51 Hz, 1 H) 7.01 (dl, J = 7.88 Hz, 1 H) 7.33 (dl, J = 7.88 Hz, 1 H) 7.44 (dl, J = 8.20 Hz, 2 H) 7.56 (dl, J = 8.20 Hz, 2 H) 8.04 (sl, 1 H) 12.01 (sl, 1 H)
[00289] [00289] LC / MS (LC-C method): Rt 3.18 min, MH + 577 14A enantiomer: 1
[00290] [00290] H NMR (500 MHz, DMSO-d6) į ppm 1.40 - 1.61 (m, 4 H) 2.14 - 2.24 (m, 2 H) 2.33 - 2.42 (m , 2 H) 3.06 - 3.23 (m, 2 H) 3.62 (s, 3 H) 3.92 - 4.18 (m, 1 H) 4.40 - 4.59 (m, 1 H) 5.40 - 5.69 (m, 1 H) 6.00 (s, 1 H) 6.13 (s, 1 H) 6.20 (s, 1 H) 6.30 - 6.47 ( m, 1 H) 6.91 - 7.12 (m, 1 H) 7.28 - 7.38 (m, 1 H) 7.44 (d, J = 8.20 Hz, 2 H) 7.56 (d, J = 8.51 Hz, 2 H) 7.95 - 8.29 (m, 1 H) 11.99 (ls, 1 H)
[00291] [00291] LC / MS (LC-C method): Rt 3.21 min, MH + 577
[00292] [00292] [Į] D20: + 55.8 ° (c 0.312, DMF)
[00293] [00293] Chiral SFC (SFC-L method): Rt 1.32 min, without MH +, 100% chiral purity. 14B Enantiomer: 1
[00294] [00294] H NMR (500 MHz, DMSO-d6) į ppm 1.40 - 1.53 (m, 4 H) 2.15 - 2.25 (m, 2 H) 2.32 - 2.44 (m , 2 H) 3.00 - 3.27 (m, 2 H) 3.62 (s, 3 H) 3.95 - 4.18 (m, 1 H) 4.44 - 4.79 (m, 1 H) 5.57 (d, J = 8.83 Hz, 1 H) 6.00 (s, 1 H) 6.13 (s, 1 H) 6.20 (s, 1 H) 6.37 (dl , J = 8.83 Hz, 1 H) 7.02 (dl, J = 7.25 Hz, 1 H) 7.34 (d, J = 8.20 Hz, 1 H) 7.44 (d, J = 8.20 Hz, 2 H) 7.56 (d, J = 8.51 Hz, 2 H) 8.04 (s, 1 H) 12.00 (m, 1 H)
[00295] [00295] LC / MS (LC-C method): Rt 3.20 min, MH + 577
[00296] [00296] [Į] D20: -53.7 ° (c 0.326, DMF)
[00297] [00297] Chiral SFC (SFC-L method): Rt 1.74 min, without MH +, 100% chiral purity. Example 15: 4- (3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenyl) butanoic acid synthesis ( Compound 15) and chiral separation in Enantiomers 15A and 15B Synthesis of intermediate 15a:
[00298] [00298] 4-Ethoxy-4-oxobutylzinc bromide (5.2 mL, 2.6 mmol) and Pd2 (dba) 3 (0.20 g, 0.22 mmol) were added to a degassed 1-bromine solution -3-methoxy-5-nitrobenzene [CAS 16618-67-0] (0.5 g, 2.2 mmol), tri-tert-butylphosphine (87 mg, 0.43 mmol) and Cs2CO3 (1.4 g, 4.3 mmol) in THF (8.8 mL). The reaction was stirred at room temperature for 18 h under N2 atmosphere. The mixture was concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel (30 µm, 80 g, heptane / EtOAc from 95/5 to 80/20) to give ethyl 4- (3-methoxy-5-nitrophenyl) butanoate 15a (370 mg ). Synthesis of intermediate 15b:
[00299] [00299] A mixture of ethyl 4- (3-methoxy-5-nitrophenyl) butanoate 15a (0.37 g, 1.38 mmol) in EtOH (8.1 mL) was hydrogenated under an atmospheric pressure of H2 for 18 h with Pd / C (10%) (0.15 g, 0.14 mmol) as a catalyst. The catalyst was removed by filtration through a pad of Celite®. Celite® was washed with EtOAc and the filtrate was concentrated in vacuo to give ethyl 4- (3-amino-5-methoxyphenyl) butanoate 15b (350 mg). The compound was used as such in the next step. Synthesis of intermediate 15c:
[00300] [00300] A mixture of 2-bromo-2- (4-chlorophenyl) -1- (6- (trifluoromethoxy) indolin-1-yl) ethanone 1c (0.43 g, 0.98 mmol), 4- (3 Ethyl-amino-5-methoxyphenyl) butanoate 15b (0.35 g, 1.48 mmol) and diisopropylethylamine (0.34 mL, 2.0 mmol) in CH3CN (5.1 mL) was stirred at 80 ° C for 18 h. The mixture was absorbed with EtO Ac and washed with 0.5 N HCl (twice) and water. The organic layer was dried over MgSO4, filtered and the solvent was evaporated in vacuo. Purification by flash chromatography on silica gel (30 µm, 40 g, heptane / EtOAc from 85/15 to 75/25) gave 4- (3 - ((1- (4-chlorophenyl) -2-oxo- Ethyl 2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenyl) butanoate 15c (340 mg). Synthesis of Compound 15 and chiral separation in Enantiomers 15A and 15B:
[00301] [00301] Under N2, a solution of LiOH monohydrate (0.12 g, 2.9 mmol) in water (3.1 mL) was added dropwise to a solution of 4- (3 - ((1- Ethyl (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenyl) butanoate 15c (0.34 g, 0.58 mmol) in THF (7 mL). The mixture was stirred at room temperature for 72 h. The solution was acidified with HCl (3 N) and extracted with EtOAc. The organic layer was dried over MgSO4, filtered and concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel (30 µm, 24 g, CH2Cl2 / MeOH 100/0 to 98/2) to give, after solidification from Et2O, 4- (3 - ((1- ( 4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenyl) butanoic (compound 15, 285 mg). The enantiomers were separated by chiral SFC (Stationary phase: Chiralcel® OD-H 5 µm 250 x 30 mm, mobile phase: 65% CO2, 35% MeOH) to give, after solidification from Et2O, the eluted Enantiomer firstly 15A (75 mg) and the eluted Enantiomer secondly 15B (85 mg). Compound 15: 1
[00302] [00302] H NMR (500 MHz, DMSO-d6) į ppm 1.66 - 1.86 (m, 2 H) 2.11 - 2.26 (m, 2 H) 2.32 - 2.44 (m , 2 H) 3.03 - 3.26 (m, 2 H) 3.62 (s, 3 H) 4.04 (td, J = 10.32, 7.09 Hz, 1 H) 4.52 ( td, J = 10.32, 6.15 Hz, 1 H) 5.56 (d, J = 8.83 Hz, 1 H) 6.00 (s, 1 H) 6.14 (s, 1 H) 6.21 (s, 1 H) 6.38 (d, J = 8.83 Hz, 1 H) 7.01 (dd, J = 8.20, 1.26 Hz, 1 H) 7.33 (d , J = 8.20 Hz, 1 H) 7.38 - 7.50 (m, 2 H) 7.56 (d, J = 8.20 Hz, 2 H) 8.03 (s, 1 H) 12 .01 (s, 1H)
[00303] [00303] LC / MS (LC-D method): Rt 2.96 min, MH + 563 15A enantiomer: 1
[00304] [00304] H NMR (500 MHz, DMSO-d6) į ppm 1.56 - 1.87 (m, 2 H) 2.11 - 2.21 (m, 2 H) 2.35 - 2.42 (m , 2 H) 3.02 - 3.22 (m, 2 H) 3.62 (s, 3 H) 3.97 - 4.18 (m, 1 H) 4.39 - 4.61 (m, 1 H) 5.56 (d, J = 8.59 Hz, 1 H) 5.99 (s, 1 H) 6.13 (s, 1 H) 6.20 (s, 1 H) 6.37 (dl , J = 9.09 Hz, 1 H) 7.00 (dl, J = 8.59 Hz, 1 H) 7.33 (d, J = 8.08 Hz, 1 H) 7.43 (d, J = 8.59 Hz, 2 H) 7.55 (d, J = 8.59 Hz, 2 H) 8.03 (s, 1 H) 11.99 (ls, 1 H)
[00305] [00305] LC / MS (LC-C method): Rt 3.08 min, MH + 563
[00306] [00306] [Į] D20: -59.0 ° (c 0.293, DMF)
[00307] [00307] Chiral SFC (SFC-M method): Rt 2.19 min, without MH +, chiral purity 99.31%. 15B Enantiomer: 1
[00308] [00308] H NMR (500 MHz, DMSO-d6) į ppm 1.60 - 1.86 (m, 2 H) 2.10 - 2.21 (m, 2 H) 2.35 - 2.42 (m , 2 H) 3.02 - 3.23 (m, 2 H) 3.62 (s, 3 H) 3.82 - 4.26 (m, 1 H) 4.36 - 4.71 (m, 1 H) 5.56 (d, J = 9.09 Hz, 1 H) 5.99 (s, 1 H) 6.13 (s, 1 H) 6.20 (s, 1 H) 6.37 (d , J = 9.09 Hz, 1 H) 7.00 (dl, J = 8.59 Hz, 1 H) 7.33 (d, J = 8.08 Hz, 1 H) 7.43 (d, J = 8.59 Hz, 2 H) 7.55 (d, J = 8.59 Hz, 2 H) 8.03 (s, 1 H) 11.97 (s, 1 H)
[00309] [00309] LC / MS (LC-C method): Rt 3.08 min, MH + 563
[00310] [00310] [Į] D20: + 48.0 ° (c 0.225, DMF)
[00311] [00311] Chiral SFC (SFC-M method): Rt 3.73 min, without MH +, chiral purity 99.61%. Example 16: synthesis of 3- (3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxybenzyl) oxide) propanoic (Compound 16) Synthesis of intermediate 16a:
[00312] [00312] tert-Butyldimethylsilyl chloride (6.1 g, 40.48 mmol) was added to a solution of (3-amino-5-methoxyphenyl) methanol [1261566- 52-2] (3.1 g, 20, 24 mmol) and imidazole (4.13 g, 60.71 mmol) in CH2Cl2 (130 mL) at room temperature. The reaction was stirred at room temperature for 16 h, quenched with water and extracted with CH2Cl2. The organic layer was separated, washed with water and brine, dried over MgSO4, filtered and evaporated to dryness. The residue was purified by flash chromatography on silica gel (30 µm, 120 g, heptane / EtOAc
[00313] [00313] A mixture of benzyl bromide (3.8 mL, 31.8 mmol), 3- (((tert-butyldimethylsilyl) oxy) -methyl) -5-methoxyaniline 16a (3.4 g, 12.71 mmol ), K2CO3 (5.27 g, 38.14 mmol) and DMAP (155 mg, 1.27 mmol) in CH3CN (66 mL) was stirred at 60 ° C for 18 h. The mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel (30 µm, 120 g, heptane / EtOAc from 100/0 to 90/10). The pure fractions were combined and evaporated to dryness to give N, N-dibenzyl-3 - (((tert-butyldimethylsilyl) oxy) methyl) -5-methoxyaniline 16b (6 g). Synthesis of intermediate 16c:
[00314] [00314] TBAF (1 M in THF) (15.3 mL, 15.3 mmol) was added to a solution of N, N-dibenzyl-3 - ((((tert-butyldimethylsilyl) oxy) methyl) -5-methoxyaniline 16b (5.69 g, 12.71 mmol) in Me-THF (64 mL). The mixture was stirred at room temperature for 2 h. The mixture was diluted with EtOAc, washed 3 times with brine and with a saturated aqueous solution of NaHCO3, dried over MgSO4, filtered and concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel (30 µm, 120 g, heptane / EtOAc from 85/15 to 75/25). The pure fractions were combined and evaporated to dryness to provide 16c (4.1 (g) (3- (dibenzylamino) -5-methoxyphenyl) methanol (4.1 g). Synthesis of intermediate 16d:
[00315] [00315] Ethyl 3-bromopropionate (0.126 mL, 0.99 mmol) was added dropwise to a solution of 16c (300 mg, 0.90 mmol) (3- (dibenzylamino) -5-methoxyphenyl) methanol (60% dispersion in mineral oil) (40 mg, 0.99 mmol) in DMF (7.0 mL). The reaction was stirred at room temperature for 16 h. After dilution with EtOAc,
[00316] [00316] A mixture of ethyl 3 - ((3- (dibenzylamino) -5-methoxybenzyl) oxy) propanoate (97 mg, 0.22 mmol) in EtOH (1.3 mL) and EtOAc (0.66 mL) ) was hydrogenated under a pressure of 7 bar H2 at room temperature for 18 h with Pd / C (10%) (24 mg, 0.022 mmol) as a catalyst. The catalyst was removed by filtration through a pad of Celite®. Celite® was washed with EtOAc and the filtrate was concentrated under reduced pressure to give ethyl 3 - ((3-amino-5-methoxybenzyl) oxy) -propanoate (52 mg). Synthesis of intermediate 16f:
[00317] [00317] A mixture of 2-bromo-2- (4-chlorophenyl) -1- (6- (trifluoromethoxy) indolin-1-yl) ethanone 1c (74 mg, 0.17 mmol), 3 - ((3- ethyl amino-5-methoxybenzyl) oxy) -propanoate 16e (52 mg, 0.21 mmol) and diisopropylethylamine (59 µL, 0.34 mmol) in CH3CN (0.89 mL) was stirred at 80 ° C for 18 h. The mixture was concentrated under reduced pressure. Purification was performed by flash chromatography on silica gel (30 µm, 12 g, heptane / EtOAc from 85/15 to 75/25). The pure fractions were combined and evaporated to dryness to give 3 - ((3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-ethyl methoxybenzyl) oxy) propanoate 16f (48 mg). Synthesis of Compound 16:
[00318] [00318] A solution of LiOH monohydrate (33 mg, 0.79 mmol) in water (0.43 mL) was added dropwise to a solution of 3 - ((3- ((1- (4-chlorophenyl) ) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-
[00319] [00319] H NMR (400 MHz, DMSO-d6) į ppm 2.34 - 2.42 (m, 2 H) 3.03 - 3.23 (m, 2 H) 3.49 - 3.59 (m , 2 H) 3.63 (s, 3 H) 3.97 - 4.21 (m, 2 H) 4.27 (s, 2 H) 4.49 - 4.57 (m, 1 H) 5, 58 (d, J = 8.59 Hz, 1 H) 6.11 (s, 1 H) 6.21 (s, 1 H) 6.33 (s, 1 H) 6.48 (dl, J = 8 , 59 Hz, 1 H) 7.00 (dl, J = 8.59 Hz, 1 H) 7.33 (d, J = 8.08 Hz, 1 H) 7.43 (d, J = 8.59 Hz, 2 H) 7.56 (d, J = 8.59 Hz, 2 H) 8.03 (s, 1 H)
[00320] [00320] LC / MS (LC-C method): Rt 2.84 min, MH + 579 Example 17: 4- (3 - ((1- (4-chloro-2-methoxyphenyl) -2-oxo- 2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) -2-methylbutanoic (Compound 17) and separation into Stereoisomers 17A, 17B, 17C and 17D.
[00321] [00321] tert-Butyl 4-bromo-2-methylbutanoate [CAS 1210410-44-8] (3.9 g, 16.446 mmol) was added dropwise to a mixture of 3-methoxy-5-nitrophenol [7145- 49-5] (2.78 g, 16.446 mmol) and K2CO3 (3.409 g, 24.669 mmol) in DMF (25 mL). The mixture was stirred at room temperature for 7 h and at 60 ° C for 16 h. After cooling to 0ºC, the mixture was diluted with water and ice and extracted with EtOAc. The organic layer was washed with water, dried over MgSO4, filtered and the solvent was evaporated to dryness. The crude residue was purified by flash chromatography on silica gel (20-45 µm, 80 g, mobile phase: heptane / EtOAc 75/25). The pure fractions were combined and the solvent evaporated under reduced pressure to give tert-butyl 4- (3-methoxy-5-nitrophenoxy) -2-methylbutanoate 17a (2.59 g). Synthesis of intermediate 17b:
[00322] [00322] A mixture of tert-butyl 4- (3-methoxy-5-nitrophenoxy) -2-methylbutanoate 17a (2.9 g, 8.913 mmol) in MeOH (50 mL) was hydrogenated under an atmospheric pressure of H2 for 4 h with Pd / C (10%) (1.52 g, 1.426 mmol) as a catalyst. The catalyst was removed by filtration through a pad of Celite®. Celite® was washed with EtOAc and the filtrate concentrated under reduced pressure. The mixture was purified by flash chromatography on silica gel (20-45 µm, 40 g, mobile phase: heptane / EtOAc 85/15). The pure fractions were combined and the solvent evaporated to dryness to give tert-butyl 4- (3-amino-5-methoxyphenoxy) -2-methylbutanoate 17b (2.29 g). Synthesis of intermediate 17c:
[00323] [00323] A mixture of 6- (trifluoromethoxy) indoline [CAS 959235-95-1] (2.5 g, 12.3 mmol), 2- (4-chloro-2-methoxyphenyl) acetic acid [CAS 170737-95 -8] (2.47 g, 12.3 mmol), HATU (7 g, 18.5 mmol) and diisopropylethylamine (6.1 mL, 36.9 mmol) in DMF (40 mL) was stirred at temperature room for 4 h. Water and EtOAc were added. The organic layer was separated, washed with water, dried over MgSO4, filtered, and the solvent evaporated under reduced pressure. The residue was purified by chromatography on silica gel (15-40 µm, 80 g, heptane / EtOAc 85/15). The pure fractions were combined and the solvent was concentrated under reduced pressure to give, after crystallization from CH3CN / heptane, 2- (4-chloro-2-methoxyphenyl) -1- (6- (trifluoromethoxy) indolin-1-yl ) ethanone 17c (4.3 g). Synthesis of intermediate 17d:
[00324] [00324] At -78 ° C, under a flow of N 2, 1 M LiHMDS in THF (19.7 mL, 19.7 mmol) was added dropwise to a mixture of 2- (4-chloro-2- methoxyphenyl) -1- (6- (trifluoromethoxy) indolin-1-yl) ethanone 17c (3.8 g, 9.8 mmol) in THF (50 mL). TMSCl (1.5 mL, 11.8 mmol) was added dropwise. The mixture was stirred for 15 min at -78 ° C and a solution of N-bromosuccinimide (1.9 g, 10.8 mmol) in THF (35 ml) was added dropwise. After stirring for 2 h at -78 ° C, the reaction was quenched with a saturated solution of NH4Cl. The mixture was extracted with EtOAc. The organic layer was separated, dried over MgSO4, filtered and the solvent evaporated under reduced pressure to give 2-bromo-2- (4-chloro-2-methoxyphenyl) -1- (6- (trifluoromethoxy) indolin-1-yl) 17d ethanone (4.5 g). The compound was used as such in the next step. Synthesis of intermediate 17e and separation in Stereoisomers 17f, 17g, 17h and 17i:
[00325] [00325] A mixture of 2-bromo-2- (4-chloro-2-methoxyphenyl) -1- (6- (trifluoromethoxy) indolin-1-yl) ethanone 17d (2 g, 4.304 mmol), 4- (3 tert-butyl -amino- 5-methoxyphenoxy) -2-methylbutanoate 17b (763 mg, 2.583 mmol) and diisopropylethylamine (1.5 mL, 8.608 mmol) in CH3CN (70 mL) was stirred at 60 ° C for 6 H. The mixture was cooled to 0 ° C, diluted with water and ice and extracted with EtOAc. The organic layer was washed with water, dried over MgSO4, filtered and the solvent was evaporated under reduced pressure. Purification was performed by flash chromatography on silica gel (25-30 µm, 40 g, heptane / EtOAc 80/20). A second purification was carried out by flash chromatography on silica gel (25-30 µm, 40 g, heptane / EtOAc 85/15). The pure fractions were combined and evaporated to dryness to give 4- (3 - ((1- (4-chloro-2-methoxyphenyl) -2-oxo- 2- (6- (trifluoromethoxy) indolin-1-yl) ethyl ) tert-butyl 17e amino) -5-methoxyphenoxy) -2-methylbutanoate (787 mg). The stereoisomers were separated by chiral SFC (Stationary phase: Chiralpak® AD-H 5 µm 250 x 30 mm, mobile phase: 83% CO2, 17% EtOH) to give a mixture of 17f and 17g (348 mg) and 17h (164 mg) and 17i (184 mg) pure. 17f and 17g were further separated by chiral SFC (Stationary phase: Chiralpak® AD-H 5 µm 250 x 30 mm, mobile phase: 88% CO2, 12% EtOH) to give 17f (145 mg) and 17g (140 mg). Synthesis of Compound 17:
[00326] [00326] A mixture of 4- (3 - ((1- (4-chloro-2-methoxyphenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5- tert-butyl methoxyphenoxy) -2-methylbutanoate 17e (180 mg, 0.265 mmol) in HCl (4 M in dioxane) (3 mL) was stirred at room temperature for 18 h. The mixture was concentrated under reduced pressure. 5 ml of triethylamine was added and the solution was evaporated in vacuo. Purification was performed by flash chromatography on silica gel (30 µm, 12 g, CH2Cl2 / MeOH from 99/1 to 96/4). The pure fractions were combined and evaporated to dryness (m = 115 mg). The residue was solidified in pentane / diisopropyl ether and a few drops of CH3CN to give 4- (3 - ((1- (4-chloro-2-methoxyphenyl) -2-oxo-2- (6- (trifluoromethoxy acid ) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) -2-methylbutanoic (Compound 17, 75 mg). Synthesis of Stereoisomer 17A:
[00327] [00327] A mixture of 17f (145 mg, 0.214 mmol) in HCl (4 M in dioxane) (5 ml) was stirred at room temperature for 6 h. The mixture was concentrated under reduced pressure. 5 ml of Et3N was added and the solution was evaporated in vacuo. Purification was performed by flash chromatography on silica gel (15-40 µm, 12 g, CH2Cl2 / MeOH from 99.5 / 0.5 to 95/5). The pure fractions were combined and evaporated to dryness (m = 93 mg). The residue was solidified from pentane / diisopropyl ether and a few drops of CH3CN to give Stereoisomer 17A (64 mg). Synthesis of Stereoisomer 17B:
[00328] [00328] A mixture of 17g (135 mg, 0.199 mmol) in HCl (4 M in dioxane) (5 ml) was stirred at room temperature for 18 h. The mixture was concentrated under reduced pressure. 5 ml of Et3N was added and the solution was evaporated in vacuo. Purification was performed by flash chromatography on silica gel (15-40 µm, 12 g, CH2Cl2 / MeOH from 99.5 / 0.5 to 95/5). The pure fractions were combined and evaporated to dryness (m = 65 mg). The residue was solidified from pentane / diisopropyl ether and a few drops of CH3CN to give Stereoisomer 17B (38 mg). Synthesis of the 17C Stereoisomer:
[00329] [00329] A mixture of 17h (162 mg, 0.239 mmol) in HCl (4 M in dioxane) (5 ml) was stirred at room temperature for 6 h. The mixture was concentrated under reduced pressure. 5 ml of Et3N was added and the solution was evaporated in vacuo. Purification was performed by flash chromatography on silica gel (15-40 µm, 12 g, CH2Cl2 / MeOH from 99.5 / 0.5 to 95/5). The pure fractions were combined and evaporated to dryness (m = 85 mg). The residue was solidified from pentane / diisopropyl ether and a few drops of CH3CN to give Stereoisomer 17C (68 mg). Synthesis of the 17D Stereoisomer:
[00330] [00330] A mixture of 17i (179 mg, 0.264 mmol) in HCl (4 M in dioxane) (5 ml) was stirred at room temperature for 6 h. The mixture was concentrated under reduced pressure. 5 ml of Et3N was added and the solution was evaporated in vacuo. Purification was performed by flash chromatography on silica gel (15-40 µm, 24 g, CH2Cl2 / MeOH from 99.5 / 0.5 to 95/5). The pure fractions were combined and evaporated to dryness (m = 98 mg). The residue was solidified from pentane / diisopropyl ether and a few drops of CH3CN to give Stereoisomer 17D (54 mg). Compound 17: 1
[00331] [00331] H NMR (500 MHz, DMSO-d6) į ppm 1.11 (dl, J = 6.62 Hz, 3 H) 1.54 - 1.82 (m, 1 H) 1.82 - 2, 07 (m, 1 H) 3.10 - 3.22 (m, 2 H) 3.62 (s, 3 H) 3.77 - 3.87 (m, 2 H) 3.91 (s, 3 H ) 3.96 - 4.18 (m, 1 H) 4.30 - 4.43 (m, 1 H) 5.60 (dl, J = 8.20 Hz, 1 H) 5.76 (ls, 1 H) 5.87 (s 1 H) 5.88 (s, 1H) 6.46 (dl, J = 8.51 Hz, 1 H) 6.98 - 7.6 (m, 2 H) 7.15 (ls, 1 H) 7.28 - 7.44 (m, 2 H) 8.03 (ls, 1 H) 12.20 (ls, 1 H) (1H not having CH CO2H under DMSO)
[00332] [00332] LC / MS (LC-C method): Rt 3.16 min, MH + 623 17A Stereoisomer: 1
[00333] [00333] H NMR (400 MHz, DMSO-d6) į ppm 1.10 (dl, J = 7.07 Hz, 3 H) 1.59 - 1.82 (m, 1 H) 1.82 - 2, 04 (m, 1 H) 3.04 - 3.24 (m, 2 H) 3.61 (s,
[00334] [00334] LC / MS (LC-D method): Rt 3.07 min, MH + 623
[00335] [00335] [Į] D20: -18.4 ° (c 0.305, DMF)
[00336] [00336] Chiral SFC (SFC-N method): Rt 4.75 min, MH + 623, chiral purity 99.3%. Stereoisomer 17B: 1
[00337] [00337] H NMR (400 MHz, DMSO-d6) į ppm 1.09 (dl, J = 6.57 Hz, 3 H) 1.64 - 1.83 (m, 1 H) 1.83 - 2, 09 (m, 1 H) 3.00 - 3.23 (m, 2 H) 3.61 (s, 3 H) 3.68 - 3.86 (m, 2 H) 3.90 (m, 3 H ) 3.96 - 4.21 (m, 1 H) 4.26 - 4.56 (m, 1 H) 5.59 (dl, J = 8.08 Hz, 1 H) 5.75 (ls, 1 H) 5.86 (s, 1H) 5.87 (s, 1 H) 6.44 (dl, J = 8.59 Hz, 1 H) 6.97 - 7.06 (m, 2 H) 7, 14 (s, 1 H) 7.26 - 7.34 (m, 2 H) 8.02 (ls, 1 H) 12.20 (ls, 1 H) (1H not having CH CO2H under DMSO)
[00338] [00338] LC / MS (LC-D method): Rt 3.03 min, MH + 623
[00339] [00339] [Į] D20: -51.0 ° (c 0.298, DMF)
[00340] [00340] Chiral SFC (SFC-N method): Rt 5.90 min, MH + 623, chiral purity 97.94%. Stereoisomer 17C: 1
[00341] [00341] H NMR (400 MHz, DMSO-d6) į ppm 1.09 (d, J = 6.57 Hz, 3 H) 1.61 - 1.84 (m, 1 H) 1.88 - 2, 02 (m, 1 H) 3.07 - 3.26 (m, 2 H) 3.61 (s, 3 H) 3.76 - 3.88 (m, 2 H) 3.90 (s, 3 H ) 3.97 - 4.18 (m, 1 H) 4.27 - 4.45 (m, 1 H) 5.59 (dl, J = 8.59 Hz, 1 H) 5.75 (s, 1 H) 5.86 (ls, 1 H) 5.87 (ls, 1 H) 6.39 - 6.49 (m, 1 H) 6.97 - 7.02 (m, 1 H) 7.02 - 7.05 (m, 1 H) 7.14 (d, J = 2.02 Hz, 1 H) 7.29 - 7.32 (m, 1 H) 7.32 - 7.37 (m, 1 H ) 8.02 (s, 1 H) 12.19 (ls, 1 H) (1H not having CH CO2H under DMSO)
[00342] [00342] LC / MS (LC-C method): Rt 3.16 min, MH + 623
[00343] [00343] [Į] D20: + 41.6 ° (c 0.257, DMF)
[00344] [00344] Chiral SFC (SFC-N method): Rt 6.86 min, MH + 623, chiral purity 98.89%. 17D Stereoisomer: 1
[00345] [00345] H NMR (500 MHz, DMSO-d6) į ppm 1.11 (d, J = 6.94 Hz, 3 H) 1.70 (dq, J = 13.44, 6.55 Hz, 1 H ) 1.97 (dq, J = 13.64, 6.80 Hz, 1 H) 3.09 -3.26 (m, 2 H) 3.62 (s, 3 H) 3.85 (tl, J = 6.31 Hz, 2 H) 3.91 (s, 3 H) 3.97 - 4.09 (m, 1 H) 4.31 - 4.47 (m, 1 H) 5.60 (dl, J = 8.51 Hz, 1 H) 5.76 (s, 1 H) 5.86 (ls, 1 H) 5.88 (ls, 1 H) 6.45 (dl, J = 8.51 Hz, 1 H) 5.76 (s, 1 H) 5.86 (ls, 1 H) 5.88 (ls, 1 H) 6.45 (dl, J = 1.26 Hz, 1 H) 7.32 ( d, J = 8.20 Hz, 1 H) 7.34 (dl, J = 8.20 Hz, 1 H) 7.34 (dl, (1H having no CH CO2H under DMSO)
[00346] [00346] LC / MS (LC-C method): Rt 3.15 min, MH + 623
[00347] [00347] [Į] D20: + 15.8 ° (c 0.297, DMF)
[00348] [00348] Chiral SFC (SFC-N method): Rt 8.14 min, MH + 623, chiral purity 98.6%. Example 18: 4- (3 - ((1- (4-chloro-2-methoxyphenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5- acid synthesis methoxyphenoxy) -2,2-dimethyl-butanoic (Compound 18) and chiral separation in Enantiomers 18A and 18B.
[00349] [00349] A mixture of 2-bromo-2- (4-chloro-2-methoxyphenyl) -1- (6- (trifluoromethoxy) indolin-1-yl) ethanone 17d (800 mg, 1.291 mmol), 4- (3 - methyl amino-5-methoxyphenoxy) -2,2-dimethylbutanoate 2a (518 mg, 1.937 mmol) and diisopropylethylamine (445 µL, 2.583 mmol) in CH3CN (6 mL) was stirred at 80 ° C for 48 h. The mixture was concentrated to dryness. The residue was absorbed with EtOAc, washed with 1 N HCl (twice) and with water. The organic layer was dried over MgSO4, filtered and the solvent was evaporated in vacuo to give 4- (3 - ((1- (4-chloro-2-methoxyphenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin Methyl -1-yl) ethyl) amino) -5-methoxyphenoxy) -2,2-dimethylbutanoate 18a (950 mg). The compound was used as such in the next step. Synthesis of Compound 18 and chiral separation in Enantiomers 18A and 18B:
[00350] [00350] At 0 ° C, LiOH monohydrate (184 mg, 4.38 mmol) was added portion by portion to a solution of 4- (3 - ((1- (4-chloro-2-methoxyphenyl) -2-oxo -2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) -2,2-methylmethylbutanoate 18a (950 mg, 1.459 mmol) in THF / MeOH / water (1/1 / 1) (30 mL). The mixture was stirred at room temperature for 16 h and then at 60 ° C for 16 h. The mixture was diluted with water and extracted with EtOAc. The aqueous layer was slowly acidified to pH 5-6 with 3 N HCl and extracted with EtOAc. The organic layers were dried over MgSO4, filtered and the solvent was evaporated in vacuo. Purification was performed by flash chromatography on silica gel (15-40 µm, 40 g, CH2Cl2 / CH3OH, from 100/0 to 99/1). The pure fractions were combined and evaporated to dryness (m = 350 mg). A small portion of the residue was crystallized from Et2O / diisopropyl ether. The precipitate was filtered off and dried to give 4- (3 - ((1- (4-chloro-2-methoxyphenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) acid) amino) -5-methoxyphenoxy) -2,2-dimethylbutanoic (Compound 18, 25 mg). The remaining amount (290 mg) was used for chiral separation. The enantiomers were separated by chiral SFC (Stationary phase: Chiralcel® OD-H 5 µm 250 x 30 mm, mobile phase: 75% CO2, 25% MeOH) to give, after solidification from heptane / diether. -isopropyl, Enantiomer eluted first 18A (68 mg) and Enantiomer eluted second 18B (70 mg). Compound 18: 1
[00351] [00351] H NMR (500 MHz, DMSO-d6) į ppm 1.14 (d, J = 2.2 Hz, 6 H) 1.87 (t, J = 7.1 Hz, 2 H) 3.09 - 3.26 (m, 2 H) 3.61 (s, 3 H) 3.80 - 3.88 (m, 2 H) 3.90 (s, 3 H) 3.98 - 4.07 (m , 1 H) 4.33 - 4.42 (m, 1 H) 5.60 (d, J = 8.5 Hz, 1 H) 5.74 (s, 1 H) 5.85 (s, 1 H ) 5.87 (s, 1 H) 6.44 (d, J = 8.5 Hz, 1 H) 6.98 - 7.06 (m, 2 H) 7.15 (d, J = 1.6 Hz, 1 H) 7.33 (dd, J = 11.7, 8.2 Hz, 2 H) 8.03 (s, 1 H) 12.23 (s, 1 H)
[00352] [00352] LC / MS (LC-C method): Rt 3.40 min, MH + 637
[00353] [00353] PF = 138 ° C 18A enantiomer: 1
[00354] [00354] H NMR (500 MHz, DMSO-d6) į ppm 1.14 (d, J = 2.2 Hz, 6 H) 1.87 (tl, J = 7.1 Hz, 2 H) 3.09 - 3.25 (m, 2 H) 3.61 (s, 3 H) 3.81 - 3.87 (m, 2 H) 3.90 (s, 3 H) 3.98 - 4.07 (m , 1 H) 4.33 - 4.42 (m, 1 H) 5.59 (d, J = 8.5 Hz, 1 H) 5.74 (s, 1 H) 5.84 (s, 1 H ) 5.87 (s, 1 H) 6.44 (dl, J = 8.5 Hz, 1 H) 6.98 - 7.06 (m, 2 H) 7.14 (d, J = 1.9 Hz, 1 H) 7.33 (dd, J = 10.7, 8.5 Hz, 2 H) 8.02 (s, 1 H) 11.94 - 12.35 (m, 1 H)
[00355] [00355] LC / MS (LC-C method): Rt 3.40 min, MH + 637
[00356] [00356] [Į] D20: -30.2 ° (c 0.315, DMF)
[00357] [00357] Chiral SFC (SFC-O method): Rt 1.31 min, without MH +, 100% chiral purity. 18B Enantiomer: 1
[00358] [00358] H NMR (500 MHz, DMSO-d6) į ppm 1.13 (d, J = 2.5 Hz, 6 H) 1.87 (t, J = 7.3 Hz, 2 H) 3.10 - 3.25 (m, 2 H) 3.61 (s, 3 H) 3.80 - 3.87 (m, 2 H) 3.90 (s, 3 H) 4.02 (td, J = 10 , 2, 7.1 Hz, 1 H) 4.33 - 4.41 (m, 1 H) 5.59 (d, J = 8.8 Hz, 1 H) 5.73 - 5.76 (m, 1 H) 5.84 (s, 1 H) 5.87 (s, 1 H) 6.44 (d, J = 8.5 Hz, 1 H) 6.97 - 7.08 (m, 2 H) 7.14 (d, J = 1.9 Hz, 1 H) 7.33 (dd, J = 11.2, 8.4 Hz, 2 H) 8.02 (s, 1 H) 11.92 - 12 , 44 (m, 1 H)
[00359] [00359] LC / MS (LC-C method): Rt 3.40 min, MH + 637
[00360] [00360] [Į] D20: + 28.0 ° (c 0.354, DMF)
[00361] [00361] Chiral SFC (SFC-O method): Rt 1.60 min, without MH +, chiral purity 99.45%. Example 19A: synthesis of (1R *, 2R *) - 2 - ((3 - ((1- (4-chloro-2-methoxyphenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1- il) ethyl) amino) -5-methoxyphenoxy) methyl) -cyclopropanecarboxylic acid (Compound 19A) and separation into Stereoisomers 19AA and 19AB.
[00362] [00362] A mixture of 2-bromo-2- (4-chloro-2-methoxyphenyl) -1- (6- (trifluoromethoxy) indolin-1-yl) ethanone 17d (0.37 g, 0.796 mmol), 2- ((3-amino-5-methoxyphenoxy) methyl) cyclopropanecarboxylatomethoxyphenoxy) methyl) cyclopropyl) (1R *, 2R *) - methyl 6d (0.317 g, 1.194 mmol) and diisopropylethylamine (0.274 mL, 1.593 mmol) in CH3CN (10 mL) was stirred at 80 ° C for 6 h. The reaction was cooled to 0 ° C and was diluted with water and ice. The mixture was extracted with EtOAc. The organic layer was washed with water, dried over MgSO4, filtered and the solvent was evaporated to dryness. The compound was purified by flash chromatography on silica gel (25-30 µm, 24 g, heptane / EtOAc 80/20). The pure fractions were combined and the solvent was removed under reduced pressure to give 2 - ((3 - ((1- (4-chloro-2-methoxyphenyl) -2-oxo-2- (6- (trifluoromethyl) indolin-1 -yl) ethyl) amino) -5-methoxyphenoxy) methyl) -
[00363] [00363] A solution of LiOH monohydrate (77 mg, 1.826 mmol) in water (5 mL) was added to a solution of 2 - ((3 - ((1- (4-chloro-2-methoxyphenyl) -2 -oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) methyl) (1R *, 2R *) cyclopropanecarboxylate - methyl 19a (237 mg, 0.365 mmol) in THF ( 5 mL). The mixture was stirred at room temperature for 18h and concentrated in vacuo. The compound was purified by flash chromatography on silica gel (20-45 µm, 24 g, CH2Cl2 / MeOH 99.5 / 0.5 to 98/2). The pure fractions were combined and concentrated under reduced pressure to give acid (1R *, 2R *) - 2 - ((3 - ((1- (4-chloro-2-methoxyphenyl) -2-oxo-2- (6- (trifluoromethyl) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) methyl) cyclopropanecarboxylic (Compound 19A, 170 mg). The stereoisomers were separated by chiral SFC (Stationary phase: Chiralcel® OD-H 5 µm 250 x 30 mm, mobile phase: 65% CO2, 35% MeOH) to give, after solidification from CH3CN / di ether -isopropyl / heptane, the Stereoisomer eluted first 19AA (67 mg) and the Stereoisomer eluted second 19AB (59 mg). Stereoisomer 19AA: 1
[00364] [00364] H NMR (500 MHz, DMSO-d6) į ppm 0.85 -0.95 (m, 1 H) 0.99 - 1.08 (m, 1 H) 1.55 (dt, J = 8 , 12, 4.30 Hz, 1 H) 1.57 - 1.67 (m, 1 H) 3.10 - 3.25 (m, 2 H) 3.61 (s, 3 H) 3.64 - 3.73 (m, 1 H) 3.83 (d ld, J = 10.40, 6.31 Hz, 1 H) 3.90 (s, 3 H) 3.98 -4.08 (m, 1 H) 4.29 - 4.44 (m, 1 H) 5.60 (dl, J = 8.51 Hz, 1 H) 5.76 (s, 1 H) 5.87 (s, 2 H) 6 , 45 (dl, J = 8.83 Hz, 1 H) 6.96 - 7.07 (m, 2 H) 7.14 (d, J = 1.26 Hz, 1 H) 7.31 (d, J = 8.51 Hz, 1 H) 7.32 - 7.38 (m, 1 H) 8.02 (sl, 1 H) 12.23 (sl, 1 H)
[00365] [00365] LC / MS (LC-D method): Rt 2.84 min, MH + 621
[00366] [00366] [Į] D20: -65.6 ° (c 0.25, DMF)
[00367] [00367] Chiral SFC (SFC-P method): Rt 1.44 min, without MH +, 100% chiral purity. Stereoisomer 19AB: 1
[00368] [00368] H NMR (500 MHz, DMSO-d6) į ppm 0.87 - 0.95 (m, 1 H) 0.99 - 1.07 (m, 1 H) 1.50 - 1.57 (m , 1 H) 1.59 - 1.70 (m, 1 H) 3.09 - 3.24 (m, 2 H) 3.61 (s, 3 H) 3.63 - 3.72 (m, 1 H) 3.85 (d ld, J = 10.40, 6.31 Hz, 1 H) 3.90 (s, 3 H) 3.99 - 4.09 (m, 1 H) 4.30 - 4 , 44 (m, 1 H) 5.60 (dl, J = 8.51 Hz, 1 H) 5.76 (s, 1 H) 5.87 (s, 2 H) 6.45 (dl, J = 8.51 Hz, 1 H) 7.00 - 7.09 (m, 2 H) 7.14 (d, J = 1.26 Hz, 1 H) 7.30 (d, J = 8.51 Hz, 1 H) 7.34 (dl, J = 8.20 Hz, 1 H) 8.02 (ls, 1 H) 12.26 (ls, 1 H)
[00369] [00369] LC / MS (LC-D method): Rt 2.85 min, MH + 621
[00370] [00370] [Į] D20: + 37.1 ° (c 0.28, DMF)
[00371] [00371] Chiral SFC (SFC-P method): Rt 2.20 min, without MH +, chiral purity 99.84%. Example 19B: synthesis of (1S *, 2S *) - 2 - ((3 - ((1- (4-chloro-2-methoxyphenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1- il) ethyl) amino) -5-methoxyphenoxy) methyl) -cyclopropanecarboxylic acid (Compound 19B) and separation into Stereoisomers 19BA and 19BB.
[00372] [00372] A mixture of 2-bromo-2- (4-chloro-2-methoxyphenyl) -1- (6- (trifluoromethoxy) indolin-1-yl) ethanone 17d (0.39 g, 0.839 mmol), 2- ((3-
[00373] [00373] A solution of LiOH monohydrate (98 mg, 2.334 mmol) in water (5 mL) was added to a solution of 2 - ((3 - ((1- (4-chloro-2-methoxyphenyl) -2 -oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) methyl) cyclopropanecarboxylate (1S *, 2S *) - methyl 19b (303 mg, 0.477 mmol) in THF ( 5 mL). The mixture was stirred at room temperature for 18 h and then concentrated in vacuo. The compound was purified by flash chromatography on silica gel (20-45 µm, 24 g, CH2Cl2 / MeOH 100/0 to 98/2). The pure fractions were combined and concentrated under reduced pressure to give acid (1S *, 2S *) - 2 - ((3 - ((1- (4-chloro-2-methoxyphenyl) -2-oxo-2- (6- (trifluoromethyl) indolin-1-yl) ethyl) amino) - 5-methoxyphenoxy) methyl) cyclopropanecarboxylic (Compound 19B, 250 mg). The stereoisomers were separated by chiral SFC (Stationary phase: Chiralcel® OD-H 5 µm 250 x 30 mm, mobile phase: 65% CO2, 35% MeOH) to give, after solidification from CH3CN / di ether -isopropyl / heptane, the Stereoisomer eluted first
[00374] [00374] H NMR (400 MHz, DMSO-d6) į ppm 0.85 - 0.95 (m, 1 H) 0.95 - 1.09 (m, 1 H) 1.54 (dt, J = 8 , 34, 4.42 Hz, 1 H) 1.55 - 1.66 (m, 1 H) 3.09 - 3.25 (m, 2 H) 3.61 (s, 3 H) 3.66 ( dd, J = 10.36, 7.33 Hz, 1 H) 3.85 (dd, J = 10.61, 6.06 Hz, 1 H) 3.89 (s, 3 H) 3.96 - 4 , 12 (m, 1 H) 4.26 - 4.43 (m, 1 H) 5.59 (d, J = 8.59 Hz, 1 H) 5.76 (s, 1 H) 5.87 ( s, 1 H) 5.88 (s, 1H) 6.44 (d, J = 9.09 Hz, 1 H) 6.93 - 7.06 (m, 2 H) 7.14 (d, J = 2.02 Hz, 1 H) 7.30 (d, J = 8.08 Hz, 1 H) 7.34 (d, J = 8.08 Hz, 1 H) 8.02 (s, 1 H) 12 , 20 (ls, 1 H)
[00375] [00375] LC / MS (LC-D method): Rt 2.84 min, MH + 621
[00376] [00376] [Į] D20: -47.6 ° (c 0.271, DMF)
[00377] [00377] Chiral SFC (SFC-P method): Rt 1.48 min, without MH +, 100% chiral purity. Stereoisomer 19BB: 1
[00378] [00378] H NMR (400 MHz, DMSO-d6) į ppm 0.85 - 0.97 (m, 1 H) 0.98 - 1.07 (m, 1 H) 1.51 - 1.58 (m , 1 H) 1.55 -1.67 (m, 1 H) 3.07 - 3.25 (m, 2 H) 3.61 (s, 3 H) 3.65 - 3.74 (m, 1 H) 3.83 (d ld, J = 10.36, 5.81 Hz, 1 H) 3.90 (s, 3 H) 3.97 - 4.14 (m, 1 H) 4.30 - 4 , 42 (m, 1 H) 5.60 (dl, J = 8.59 Hz, 1 H) 5.76 (s, 1 H) 5.87 (s, 2 H) 6.44 (dl, J = 8.59 Hz, 1 H) 6.98 - 7.07 (m, 2 H) 7.14 (d, J = 1.52 Hz, 1 H) 7.30 (d, J = 8.08 Hz, 1 H) 7.33 (dl, J = 8.08 Hz, 1 H) 8.02 (s, 1 H) 12.24 (ls, 1 H)
[00379] [00379] LC / MS (LC-D method): Rt 2.84 min, MH + 621
[00380] [00380] [Į] D20: + 56.8 ° (c 0.264, DMF)
[00381] [00381] Chiral SFC (SFC-P method): Rt 2.12 min, without MH +, chiral purity 99.59%. Example 20: 4- (3 - ((1- (4-chlorophenyl) -2- (5-methoxy-6- (trifluoro-methyl) indolin-1-yl) -2-oxoethyl) amino) -5 acid synthesis -methoxyphenoxy) -2- methylbutanoic (Compound 20) and separation into Stereoisomers 20A,
[00382] [00382] A mixture of 1-methoxy-4-nitro-2- (trifluoromethyl) benzene [CAS 654-76-2] (24.5 g, 110.8 mmol) and 4-chlorophenoxyacetonitrile [CAS 3598-13-8 ] (20.4 g, 121.9 mmol) in DMF (100 mL) was added dropwise over 30 min to a stirred solution of tBuOK (27.35 g, 243.7 mmol) in DMF (100 mL) at -10 ° C. After addition, the purple solution was maintained at -10 ° C for 1 h. 500 ml of ice water and 500 ml of 6 N HCl were added and the precipitate was filtered off, washed with water and dried under reduced pressure to give 40.4 g of 2- (5-methoxy-2-nitro-4 - (trifluoromethyl) phenyl) acetonitrile 20a (used as such in the next step). Synthesis of intermediate 20b:
[00383] [00383] A solution of 2- (5-methoxy-2-nitro-4- (trifluoromethyl) phenyl) acetonitrile 20a (26 g, 99.9 mmol) in ethanol / water (9/1) (500 mL) and AcOH (5.2 mL) was hydrogenated for 1 h at a pressure of 3.5 Bar with 10% Pd / C (15.3 g) as the catalyst. The reaction mixture was filtered through a pad of Celite® and the filter cake was washed with a solvent mixture of CH2Cl2 and CH3OH. The combined filtrates were concentrated under reduced pressure. The residue was filtered through a glass filter loaded with silica 60-200 µm and using heptane / EtOAc 80/20 as the eluent. Fractions containing the expected compound were combined and the solvent was concentrated under reduced pressure to give 5-methoxy-6- (trifluoromethyl) - 1H-indole 20b (15.6 g). Synthesis of intermediate 20c:
[00384] [00384] At 0 ° C, BH 3-Pyridine (23.5 mL, 232.4 mmol) was added dropwise to a solution of 5-methoxy-6- (trifluoromethyl) -1H-indole 20b (10 g, 46.5 mmol) in EtOH (60 mL). 6 N HCl (140 mL) was added slowly while maintaining the temperature below 10 ° C. The mixture was stirred at 0 ° C for 2 h. Water (200 mL) was added and the mixture was basified to pH 8-9 with a concentrated aqueous solution of NaOH (the reaction temperature was kept below 20 ° C). The precipitate was filtered off, washed with water (twice) and coevaporated under reduced pressure with toluene to give 5c-methoxy-6- (trifluoromethyl) indoline 20c (9 g). Synthesis of Compound 20 and separation into Stereoisomers 20A, 20B, 20C and 20D:
[00385] [00385] Compound 20 (330 mg) was obtained following the procedures described for the synthesis of Compound 1 starting from intermediate 20c. The 4 Stereoisomers 20A (50 mg), 20B (18 mg), 20C (68 mg) and 20D (32 mg) were obtained, in this order of elution, through two subsequent chiral SFC separations: (Stationary phase: Chiralpak® AD -H 5 µm 250 x 30 mm, mobile phase: 40% CO2, iPrOH 60%) e (Stationary phase: Chiralcel® OJ-H 5 µm 250 x 20 mm, mobile phase: 60% CO2, MeOH 40 %); followed by individual purification by flash chromatography on silica gel (15-40 µm, 12 g, CH2Cl2 / MeOH 99.5 / 0.5 to 90/10) and subsequent solidification from CH3CN / diisopropyl ether / heptane . Stereoisomer 20A:
[00386] [00386] H NMR (400 MHz, DMSO-d6) į ppm 1.10 (d, J = 7.07 Hz, 3 H) 1.60 - 1.72 (m, 1 H) 1.90 - 2, 01 (m, 1 H) 3.20 - 3.32 (m, 2 H) 3.61 (s, 3 H) 3.79 - 3.90 (m, 5 H) 3.93 - 4.09 ( m, 1 H) 4.42 - 4.53 (m, 1 H) 5.53 (dl, J = 8.59 Hz, 1 H) 5.74 (s, 1 H) 5.93 (sl, 1 H) 5.95 (ls, 1 H) 6.38 (dl, J = 9.09 Hz, 1 H) 7.23 (s, 1 H) 7.43 (dl, J = 8.08 Hz, 2 H) 7.55 (dl, J = 8.08 Hz, 2 H) 7.55 (dl, (1H not having CH CO2H under DMSO)
[00387] [00387] LC / MS (LC-C method): Rt 2.95 min, MH + 607
[00388] [00388] [Į] D20: -40.9 ° (c 0.257, DMF)
[00389] [00389] Chiral SFC (SFC-Q method): Rt 1.07 min, MH + 607, chiral purity 100%. Stereoisomer 20B: 1
[00390] [00390] H NMR (400 MHz, DMSO-d6) į ppm 1.09 (d, J = 6.94 Hz, 3 H) 1.69 (dq, J = 13.52, 6.53 Hz, 1 H ) 1.88 - 2.11 (m, 1 H) 3.08 - 3.28 (m, 2 H) 3.53 - 3.66 (m, 3 H) 3.79 - 3.90 (m, 5 H) 3.92 - 4.11 (m, 1 H) 3.92 - 4.11 (m, 1 H) 4.32 - 4.67 (m, 1 H) 5.54 (d, J = 8.51 Hz, 1 H) 5.75 (s, 1 H) 5.93 (s, 1 H) 5.95 (s, 1H) 6.30 - 6.45 (m, 1 H) 7.23 (s, 1 H) 7.43 (d, J = 8.20 Hz, 2 H) 7.55 (d, J = 8.20 Hz, 2 H) 7.55 (d, (1H not having CH CO2H under DMSO)
[00391] [00391] LC / MS (LC-C method): Rt 2.95 min, MH + 607
[00392] [00392] [Į] D20: -50.0 ° (c 0.266, DMF)
[00393] [00393] Chiral SFC (SFC-Q method): Rt 1.07 min, MH + 607, chiral purity 100%. Stereoisomer 20C: 1
[00394] [00394] H NMR (400 MHz, DMSO-d6) į ppm 1.09 (dl, J = 6.94 Hz, 3 H) 1.52 - 1.83 (m, 1 H) 1.86 - 2, 06 (m, 1 H) 3.07 - 3.28 (m, 2 H) 3.61 (s, 3 H) 3.73 - 3.91 (m, 5 H) 3.94 - 4.04 ( m, 1 H) 4.37 - 4.58 (m, 1 H) 5.54 (dl, J = 8.51 Hz, 1 H) 5.75 (s, 1 H) 5.93 (s, 1 H) 5.95 (s, 1H) 6.39 (dl, J = 8.51 Hz, 1 H) 5.75 (s, 1 H) 5.93 (s, 1 H) 5.95 (s, 1H) 6.39 (dl, J = 8.20 Hz, 2 H) 7.55 (dl, J = 8.20 Hz, 2 H) 7.55 (dl, (1H not having CH CO2H under DMSO)
[00395] [00395] LC / MS (LC-C method): Rt 2.95 min, MH + 607
[00396] [00396] [Į] D20: + 26.0 ° (c 0.288, DMF)
[00397] [00397] Chiral SFC (SFC-Q method): Rt 1.56 min, MH + 607, chiral purity 99.68%. Stereoisomer 20D: 1
[00398] [00398] H NMR (400 MHz, DMSO-d6) į ppm 1.09 (d, J = 7.07 Hz, 3 H) 1.60 - 1.75 (m, 1 H) 1.85 - 1, 99 (m, 1 H) 3.11 - 3.27 (m, 2 H) 3.61 (s, 3 H) 3.77 - 3.91 (m, 5 H) 3.93 - 4.05 ( m, 1 H) 4.44 - 4.56 (m, 1 H) 5.54 (dl, J = 8.59 Hz, 1 H) 5.74 (s, 1 H) 5.93 (sl, 1 H) 5.95 (ls, 1 H) 6.38 (dl, J = 8.59 Hz, 1 H) 5.74 (s, 1 H) 5.93 (ls, 1 H) 5.95 (ls , 1 H) 6.38 (dl, J = 8.59 Hz, 2 H) 7.55 (dl, J = 8.08 Hz, 2 H) 8.33 (s, 1 H) 12.22 (sl , 1H) (1H having no CH CO2H under DMSO)
[00399] [00399] LC / MS (LC-C method): Rt 2.96 min, MH + 607
[00400] [00400] [Į] D20: + 57.4 ° (c 0.27, DMF)
[00401] [00401] Chiral SFC (SFC-Q method): Rt 2.19 min, MH + 607, 100% chiral purity. Example 21: 4- (3 - ((1- (4-chlorophenyl) -2- (5-methoxy-6- (trifluoro-methyl) indolin-1-yl) -2-oxoethyl) amino) -5 acid synthesis -methoxyphenoxy) -2,2-dimethylbutanoic (Compound 21) and chiral separation in Enantiomers 21A and 21B Synthesis of intermediate 21a:
[00402] [00402] A mixture of 2-bromo-2- (4-chlorophenyl) -1- (5-methoxy-6-
[00403] [00403] At 0ºC, LiOH monohydrate (254 mg, 6.046 mmol) was added to a solution of 4- (3 - ((1- (4-chlorophenyl) -2- (5-methoxy-6- (trifluoromethyl) methyl indolin-1-yl) -2-oxoethyl) amino) -5-methoxyphenoxy) -2,2-dimethylbutanoate 21a (1.28 g, 2.015 mmol) in THF / MeOH / water (15 mL). The mixture was warmed to room temperature and stirred for 48 h. The mixture was cooled to 0 ° C and water was added. The mixture was acidified to pH 4-5 with 3 N HCl and extracted with EtOAc. The combined organic layers were dried over MgSO4, filtered and the solvent was evaporated. Purification was performed by flash chromatography on silica gel (15-40 µm, 40 g, CH2Cl2 / CH3OH: 100/0 to 98/2). Fractions containing expected compound were combined and evaporated to dryness. A second purification was carried out by reverse phase (stationary phase: YMC-actus Triart-C18 10 µm 30 x 150 mm, mobile phase: gradient of 70% aqueous NH4HCO3, 0.2%, 30% CH3CN to aqueous NH4HCO3) 0%, 0.2%, 100% CH3CN). The pure fractions were combined and concentrated in vacuo to give 4- (3 - ((1- (4-chlorophenyl) -2- (5-methoxy-6- (trifluoromethyl) indolin-1-yl) -2-oxoethyl) acid amino) -5-methoxyphenoxy) - 2,2-dimethylbutanoic (Compound 21, 455 mg). The enantiomers were separated by chiral SFC (Stationary phase: Chiralcel® OD-H 5 µm 250 x 30 mm, mobile phase: 55% CO2, 45% MeOH) to give, after solidification from heptane / diether. -isopropyl, Enantiomer eluted first 21A (106 mg) and Enantiomer eluted second 21B (103 mg). Compound 21: 1
[00404] [00404] H NMR (500 MHz, DMSO-d6) į ppm 1.13 (d, J = 3.8 Hz, 6 H) 1.87 (t, J = 7.3 Hz, 2 H) 3.14 - 3.30 (m, 2 H) 3.61 (s, 3 H) 3.84 (m, 5 H) 3.98 (td, J = 10.4, 7.3 Hz, 1 H) 4, 51 (td, J = 10.3, 6.1 Hz, 1 H) 5.52 (d, J = 8.5 Hz, 1 H) 5.73 (t, J = 1.9 Hz, 1 H) 5.92 (s, 1 H) 5.94 (s, 1 H) 6.39 (d, J = 8.5 Hz, 1 H) 7.23 (s, 1 H) 7.43 (d, J = 8.5 Hz, 2 H) 7.55 (d, J = 8.5 Hz, 2 H) 8.33 (s, 1 H) 12.23 (sl, 1 H)
[00405] [00405] LC / MS (LC-C method): Rt 3.19 min, MH + 621 21A enantiomer: 1
[00406] [00406] H NMR (500 MHz, DMSO-d6) į ppm 1.08 - 1.15 (m, 6 H) 1.85 (t, J = 7.3 Hz, 2 H) 3.13 - 3, 30 (m, 2 H) 3.55 - 3.65 (m, 3 H) 3.80 - 3.89 (m, 5 H) 3.98 (td, J = 10.4, 7.3 Hz, 1 H) 4.52 (td, J = 10.4, 6.3 Hz, 1 H) 5.54 (d, J = 8.8 Hz, 1 H) 5.73 (t, J = 1.9 Hz, 1 H) 5.92 (s, 1 H) 5.95 (s, 1 H) 6.38 (d, J = 8.5 Hz, 1 H) 7.23 (s, 1 H) 7, 43 (d, J = 8.5 Hz, 2 H) 7.56 (d, J = 8.5 Hz, 2 H) 8.34 (s, 1 H)
[00407] [00407] LC / MS (LC-C method): Rt 3.21 min, MH + 621
[00408] [00408] [Į] D20: -41.7 ° (c 0.254, DMF)
[00409] [00409] Chiral SFC (SFC-H method): Rt 1.23 min, MH + 621, chiral purity 100%. 21B Enantiomer: 1
[00410] [00410] H NMR (500 MHz, DMSO-d6) į ppm 1.08 - 1.16 (m, 6 H) 1.86 (t, J = 7.1 Hz, 2 H) 3.15 - 3, 29 (m, 2 H) 3.61 (s, 3 H) 3.80 - 3.90 (m, 5 H) 3.98 (td, J = 10.2, 7.3 Hz, 1 H) 4 , 52 (td, J = 10.4, 6.3 Hz, 1 H) 5.53 (d, J = 8.5 Hz, 1 H) 5.70 - 5.75 (m, 1 H) 5, 92 (s, 1 H) 5.95 (s, 1 H) 6.38 (d, J = 8.5 Hz, 1 H) 7.23 (s, 1 H) 7.43 (d, J = 8 , 5 Hz, 2 H) 7.56 (d, J = 8.5 Hz, 2
[00411] [00411] LC / MS (LC-C method): Rt 3.21 min, MH + 621
[00412] [00412] [Į] D20: + 44.0 ° (c 0.275, DMF)
[00413] [00413] Chiral SFC (SFC-H method): Rt 2.38 min, MH + 621, 100% chiral purity. Example 22A: synthesis of (1R *, 2R *) - 2 - ((3 - ((1- (4-chlorophenyl) -2- (5-methoxy-6- (trifluoromethyl) indolin-1-yl) -2 -oxoethyl) amino) -5-methoxyphenoxy) methyl) -cyclopropanecarboxylic acid (Compound 22A) and separation in Stereoisomers 22AA and 22AB Synthesis of Compound 22A and separation in Stereoisomers 22AA and 22AB:
[00414] [00414] Compound 22A (284 mg) was synthesized from intermediate 20e using the procedures described for the synthesis of Compound 6A. The two stereoisomers were separated by chiral SFC (Stationary phase: Chiralcel® OD-H 5 µm 250 x 30 mm, mobile phase: 50% CO2, 50% EtOH) to give, after solidification from pentane / ether of diisopropyl, the Stereoisomer eluted first 22AA (79 mg) and the Stereoisomer eluted second 22AB (74 mg). Stereoisomer 22AA: 1
[00415] [00415] H NMR (500 MHz, DMSO-d6) į ppm 0.85 - 0.93 (m, 1 H) 1.00 - 1.08 (m, 1 H) 1.54 (dt, J = 8 , 12, 4.30 Hz, 1 H) 1.60 -1.68 (m, 1 H) 3.12 - 3.26 (m, 2 H) 3.61 (s, 3 H) 3.68 ( dd, J = 10.40, 7.57 Hz, 1 H) 3.79 -
[00416] [00416] LC / MS (LC-C method): Rt 2.80 min, MH + 605
[00417] [00417] [Į] D20: -75.0 ° (c 0.3, DMF)
[00418] [00418] Chiral SFC (SFC-R method): Rt 0.86 min, without MH +, 100% chiral purity. Stereoisomer 22AB: 1
[00419] [00419] H NMR (500 MHz, DMSO-d6) į ppm 0.88 (tl, J = 9.30 Hz, 1 H) 1.02 (dt, J = 8.43, 4.45 Hz, 1 H ) 1.47 - 1.58 (m, 1 H) 1.59 - 1.68 (m, 1 H) 3.13 - 3.28 (m, 2 H) 3.61 (s, 3 H) 3 , 64 - 3.72 (m, 1 H) 3.84 (s, 4 H) 3.92 - 4.06 (m, 1 H) 4.50 (td, J = 10.32, 6.15 Hz , 1 H) 5.55 (d, J = 8.83 Hz, 1 H) 5.75 (s, 1 H) 5.95 (s, 1 H) 5.96 (s, 1 H) 6.39 (d, J = 8.83 Hz, 1 H) 7.23 (s, 1 H) 7.43 (d, J = 8.51 Hz, 2 H) 7.55 (d, J = 8.51 Hz , 2 H) 8.33 (s, 1 H) 12.08 (ls, 1 H)
[00420] [00420] LC / MS (LC-D method): Rt 2.69 min, MH + 605
[00421] [00421] [Į] D20: + 10.0 ° (c 0.281, DMF)
[00422] [00422] Chiral SFC (SFC-R method): Rt 1.84 min, without MH +, 100% chiral purity. Example 22B: synthesis of (1S *, 2S *) - 2 - ((3 - ((1- (4-chlorophenyl) -2- (5-methoxy-6- (trifluoromethyl) indolin-1-yl) -2 -oxoethyl) amino) -5-methoxyphenoxy) methyl) -cyclopropanecarboxylic (Compound 22B) and separation in Stereoisomers 22BA and 22BB
[00423] [00423] Compound 22B (257 mg) was synthesized from intermediate 20e using the procedure described for the synthesis of Compound 6B. The two stereoisomers were separated by chiral SFC (Stationary phase: Chiralcel® OD-H 5 µm 250 x 30 mm, mobile phase: 50% CO2, 50% EtOH) to give, after solidification from pentane / ether of diisopropyl, the stereoisomer eluted first 22BA (49 mg) and the stereoisomer eluted second 22BB (61 mg). Stereoisomer 22BA: 1
[00424] [00424] H NMR (500 MHz, DMSO-d6) į ppm 0.83 - 1.00 (m, 1 H) 0.98 - 1.09 (m, 1 H) 1.54 (dt, J = 8 , 35, 4.33 Hz, 1 H) 1.58 - 1.70 (m, 1 H) 3.13 - 3.28 (m, 2 H) 3.61 (s, 3 H) 3.68 ( dd, J = 10.09, 7.57 Hz, 1 H) 3.78 - 3.89 (m, 4 H) 3.95 - 4.05 (m, 1 H) 4.50 (td, J = 10.25, 6.31 Hz, 1 H) 5.55 (d, J = 8.83 Hz, 1 H) 5.75 (s, 1 H) 5.95 (ls, 1 H) 5.96 ( sl, 1 H) 6.39 (dl, J = 8.83 Hz, 1 H) 7.23 (s, 1 H) 7.43 (d, J = 8.51 Hz, 2 H) 7.55 ( d, J = 8.51 Hz, 2 H) 8.33 (s, 1 H) 12.24 (sl, 1 H)
[00425] [00425] LC / MS (LC-D method): Rt 2.68 min, MH + 605
[00426] [00426] [Į] D20: -9.3 ° (c 0.291, DMF)
[00427] [00427] Chiral SFC (SFC-S method): Rt 1.48 min, MH + 605, chiral purity 100%.
[00428] [00428] H NMR (500 MHz, DMSO-d6) į ppm 0.82 - 0.95 (m, 1 H) 1.04 - 1.06 (m, 1 H) 1.55 (dt, J = 8 , 28, 4.22 Hz, 1 H) 1.58 - 1.68 (m, 1 H) 3.13 - 3.28 (m, 2 H) 3.61 (s, 3 H) 3.68 ( dd, J = 10.40, 7.25 Hz, 1 H) 3.81 - 3.88 (m, 4 H) 3.95 - 4.02 (m, 1 H) 4.46 - 4.55 ( m, 1 H) 5.54 (d, J = 8.83 Hz, 1 H) 5.75 (s, 1 H) 5.94 (s, 1 H) 5.96 (s, 1 H) 6, 39 (d, J = 9.14 Hz, 1 H) 7.23 (s, 1 H) 7.43 (d, J = 8.51 Hz, 2 H) 7.55 (d, J = 8.51 Hz, 2 H) 8.33 (s, 1 H) 12.20 (ls, 1 H)
[00429] [00429] LC / MS (LC-C method): Rt 2.80 min, MH + 605
[00430] [00430] [Į] D20: + 80.0 ° (c 0.275, DMF)
[00431] [00431] Chiral SFC (SFC-S method): Rt 3.12 min, MH + 605, chiral purity 99.55%. Example 23: synthesis of (1s, 3s) -3- (3 - ((1- (4-chlorophenyl) -2- (5-methoxy-6- (trifluoromethyl) indolin-1-yl) -2-oxoethyl) -2 amino) -5- methoxyphenoxy) cyclobutene-carboxylic (Compound 23) and chiral separation in Enantiomers 23A and 23B Synthesis of Compound 23 and chiral separation in Enantiomers 23A and 23B:
[00432] [00432] Compound 23A (280 mg) was synthesized from intermediate 20e using the procedures described for the synthesis of Compound 8. The two enantiomers were separated by chiral SFC (Stationary phase: Chiralcel® OD-H 5 µm 250 x 30 mm, mobile phase: 45% CO2, 55% EtOH) to give, after solidification from heptane / diisopropyl ether, the first eluted Enantiomer 23A (60 mg) and the second eluted Enantiomer 23B (71 mg). Compound 23: 1
[00433] [00433] H NMR (500 MHz, DMSO-d6) į ppm 2.02 - 2.15 (m, 2 H) 2.58 - 2.65 (m, 2 H) 2.66 - 2.75 (m , 1 H) 3.14 - 3.29 (m, 2 H) 3.61 (s, 3 H) 3.84 (s, 3 H) 3.98 (td, J = 10.4, 7.3 Hz, 1 H) 4.42 - 4.57 (m, 2 H) 5.51 (d, J = 8.5 Hz, 1 H) 5.65 (t, J = 1.9 Hz, 1 H) 5.86 (s, 1 H) 5.93 (s, 1 H) 6.42 (d, J = 8.5 Hz, 1 H) 7.23 (s, 1 H) 7.44 (d, J = 8.5 Hz, 2 H) 7.55 (d, J = 8.5 Hz, 2 H) 8.34 (s, 1 H) 12.11 - 12.40 (m, 1 H)
[00434] [00434] LC / MS (LC-C method): Rt 2.76 min, MH + 605 23A enantiomer: 1
[00435] [00435] H NMR (500 MHz, DMSO-d6) į ppm 2.02 - 2.15 (m, 2 H) 2.57 - 2.65 (m, 2 H) 2.66 - 2.75 (m , 1 H) 3.14 - 3.28 (m, 2 H) 3.61 (s, 3 H) 3.84 (s, 3 H) 3.98 (td, J = 10.3, 7.1 Hz, 1 H) 4.42 - 4.56 (m, 2 H) 5.51 (d, J = 8.5 Hz, 1 H) 5.65 (t, J = 2.0 Hz, 1 H) 5.86 (s, 1 H) 5.93 (s, 1 H) 6.42 (d, J = 8.5 Hz, 1 H) 7.23 (s, 1 H) 7.44 (d, J = 8.5 Hz, 2 H) 7.55 (d, J = 8.5 Hz, 2 H) 8.34 (s, 1 H) 12.02 - 12.49 (m, 1 H)
[00436] [00436] LC / MS (LC-C method): Rt 2.75 min, MH + 605
[00437] [00437] [Į] D20: -38.1 ° (c 0.307, DMF)
[00438] [00438] Chiral SFC (SFC-R method): Rt 0.84 min, MH + 605, chiral purity 100%. 23B Enantiomer: 1
[00439] [00439] H NMR (500 MHz, DMSO-d6) į ppm 2.02 - 2.15 (m, 2H) 2.56 - 2.75 (m, 3 H) 3.14 - 3.27 (m, 2 H) 3.61 (s, 3 H) 3.84 (s, 3 H) 3.93 - 4.04 (m, 1 H) 4.43 - 4.57 (m, 2 H) 5.51 (dl, J = 8.5 Hz, 1 H) 5.65 (s, 1 H) 5.86 (s, 1 H) 5.93 (s, 1 H) 6.42 (dl, J = 8, 5 Hz, 1 H) 7.23 (s, 1 H) 7.44 (dl, J = 8.5 Hz, 2 H) 7.55 (dl, J = 8.5 Hz, 2 H) 8.34 (s, 1 H) 12.07 - 12.47 (m, 1 H)
[00440] [00440] LC / MS (LC-C method): Rt 2.76 min, MH + 605
[00441] [00441] [Į] D20: + 36.9 ° (c 0.309, DMF)
[00442] [00442] Chiral SFC (SFC-R method): Rt 1.86 min, MH + 605, chiral purity 100%. Example 24: synthesis of (1s, 3s) -3 - ((3 - ((1- (4-chlorophenyl) -2- (5-methoxy-6- (trifluoromethyl) indolin-1-yl) -2-oxoethyl acid ) amino) -5-methoxyphenoxy) methyl) -cyclobutanecarboxylic (Compound 24) and chiral separation in Enantiomers 24A and 24B Synthesis of Compound 24 and chiral separation in Enantiomers 24A and 24B:
[00443] [00443] Compound 24 (550 mg) was synthesized from intermediate 20e using the procedures described for the synthesis of Compound 10. The two enantiomers were separated by chiral SFC (Stationary phase: Whelk® O1 (S, S) 5 µm 250 x 21.1 mm, mobile phase: 50% CO2, 50% MeOH) to give, after solidification from Et2O, the Enantiomer eluted first 24A (190 mg) and the Enantiomer eluted second 24B (177 mg). Compound 24: 1
[00444] [00444] H NMR (500 MHz, DMSO-d6) į ppm 1.93 (lq, J = 9.77 Hz, 2 H) 2.11 - 2.32 (m, 2 H) 2.53 - 2, 60 (m, 1 H) 2.97 (quin, J = 8.91 Hz, 1 H) 3.16 - 3.30 (m, 2 H) 3.62 (s, 3 H) 3.74 - 3 , 82 (m, 2 H) 3.85 (s, 3 H) 3.96 - 4.06 (m, 1 H) 4.51 (td, J = 10.25, 5.99 Hz, 1 H) 5.55 (d, J = 8.83 Hz, 1 H) 5.74 (s, 1 H) 5.94 (s, 2 H) 6.39 (d, J = 8.51 Hz, 1 H) 7.23 (s, 1 H) 7.44 (d, J = 8.51 Hz, 2 H) 7.55 (d, J = 8.51 Hz, 2 H) 8.34 (s, 1 H) 12.07 (ls, 1 H)
[00445] [00445] LC / MS (LC-C method): Rt 2.91 min, MH + 619
[00446] [00446] H NMR (500 MHz, DMSO-d6) į ppm 1.87 - 2.02 (m, 2 H) 2.15 - 2.30 (m, 2 H) 2.52 - 2.59 (m , 1 H) 2.95 (qt J = 8.83 Hz, 1 H) 3.07 - 3.29 (m, 2 H) 3.61 (s, 3 H) 3.77 (dl, J = 6 , 31 Hz, 2 H) 3.84 (s, 3 H) 3.95 - 4.07 (m, 1 H) 4.42 - 4.56 (m, 1 H) 5.54 (d, J = 8.83 Hz, 1 H) 5.74 (t, J = 2.05 Hz, 1 H) 5.81 - 6.01 (m, 2 H) 6.38 (d, J = 8.83 Hz, 1 H) 7.23 (s, 1 H) 7.43 (d, J = 8.51 Hz, 2 H) 7.55 (d, J = 8.51 Hz, 2 H) 8.33 (s, 1 H) 12.11 (ls, 1 H)
[00447] [00447] LC / MS (LC-C method): Rt 2.89 min, MH + 619
[00448] [00448] [Į] D20: -41.5 ° (c 0.224, DMF)
[00449] [00449] Chiral SFC (SFC-T method): Rt 1.81 min, without MH +, 100% chiral purity. 24B Enantiomer: 1
[00450] [00450] H NMR (500 MHz, DMSO-d6) į ppm 1.91 (q, J = 9.35 Hz, 2 H) 2.17 - 2.26 (m, 2 H) 2.53 - 2, 61 (m, 1 H) 2.94 (quin, J = 8.91 Hz, 1 H) 3.13 - 3.27 (m, 2 H) 3.61 (s, 3 H) 3.72 - 3 , 79 (m, 2 H) 3.84 (s, 3 H) 3.90 - 4.06 (m, 1 H) 4.50 (td, J = 10.32, 6.46 Hz, 1 H) 5.54 (d, J = 8.83 Hz, 1 H) 5.72 - 5.75 (m, 1 H) 5.91 - 5.95 (m, 2 H) 6.38 (d, J = 8.83 Hz, 1 H) 7.23 (s, 1 H) 7.43 (d, J = 8.51 Hz, 2 H) 7.55 (d, J = 8.20 Hz, 2 H) 8 , 33 (s, 1 H) 12.07 (ls, 1 H)
[00451] [00451] LC / MS (LC-C method): Rt 2.89 min, MH + 619
[00452] [00452] [Į] D20: + 36.6 ° (c 0.232, DMF)
[00453] [00453] Chiral SFC (SFC-T method): Rt 2.26 min, without MH +, chiral purity 98.71%. Example 25: synthesis of (1r, 3r) -3 - ((3 - ((1- (4-chlorophenyl) -2- (5-methoxy-6- (trifluoromethyl) indolin-1-yl) -2-oxoethyl acid ) amino) -5-methoxyphenoxy) methyl) -cyclobutanecarboxylic (Compound 25) and chiral separation in Enantiomers 25A and 25B
[00454] [00454] Compound 25 (310 mg) was synthesized from intermediate 20e using the procedures described for the synthesis of Compound 11. The two enantiomers were separated by chiral SFC (Stationary phase: Whelk® O1 (S, S) 5 µm 250 x 21.1 mm, mobile phase: 50% CO2, 50% MeOH) to give, after solidification from Et2O / pentane, the first eluted Enantiomer 25A (94 mg) and the second eluted Enantiomer place 25B (105 mg). Compound 25: 1
[00455] [00455] H NMR (400 MHz, DMSO-d6) į ppm 1.80 - 2.04 (m, 2 H) 2.22 - 2.31 (m, 2 H) 2.56 - 2.64 (m , 1 H) 3.09 (l qt, J = 7.33 Hz, 1 H) 3.14 - 3.28 (m, 2 H) 3.62 (s, 3 H) 3.84 (s, 3 H) 3.87 (d, J = 6.57 Hz, 2 H) 3.93 - 4.06 (m, 1 H) 4.45 - 4.56 (m, 1 H) 5.54 (d, J = 8.59 Hz, 1 H) 5.77 (s, 1 H) 5.94 (s, 1 H) 5.96 (s, 1 H) 6.38 (d, J = 9.09 Hz, 1 H) 7.23 (s, 1H) 7.43 (d, J = 8.59 Hz, 2 H) 7.55 (d, J = 8.59 Hz, 2 H) 8.33 (s, 1 H) 12.10 (ls, 1 H)
[00456] [00456] LC / MS (LC-C method): Rt 2.87 min, MH + 619 25A enantiomer: 1
[00457] [00457] H NMR (400 MHz, DMSO-d6) į ppm 1.91 - 2.03 (m, 2 H) 2.18 - 2.29 (m, 2 H) 2.55 - 2.62 (m , 1 H) 3.01 - 3.11 (m, 1 H) 3.14 - 3.28 (m, 2 H) 3.62 (s, 3 H) 3.84 (s, 3 H) 3, 86 (d, J = 7.07 Hz, 2 H) 3.93 - 4.05 (m, 1 H) 4.42 - 4.58 (m, 1 H) 5.54 (d, J = 8, 59 Hz, 1 H) 5.77 (s, 1 H) 5.92 - 5.95 (m, 1 H) 5.95 - 5.98 (m, 1 H) 6.38 (d, J = 8 , 59 Hz, 1 H) 7.23 (s, 1 H)
[00458] [00458] LC / MS (LC-C method): Rt 2.90 min, MH + 619
[00459] [00459] [Į] D20: -41.1 ° (c 0.28, DMF)
[00460] [00460] Chiral SFC (SFC-T method): Rt 1.91 min, without MH +, 100% chiral purity. 25B Enantiomer: 1
[00461] [00461] H NMR (400 MHz, DMSO-d6) į ppm 1.90 - 2.04 (m, 2 H) 2.19 - 2.29 (m, 2 H) 2.55 - 2.60 (m , 1 H) 3.00 - 3.30 (m, 3 H) 3.62 (s, 3 H) 3.84 (s, 3 H) 3.86 (dl, J = 7.07 Hz, 2 H ) 3.94 - 4.04 (m, 1 H) 4.45 - 4.55 (m, 1 H) 5.54 (d, J = 8.59 Hz, 1 H) 5.77 (s, 1 H) 5.88 - 5.95 (m, 1 H) 5.95 - 5.98 (m, 1 H) 6.38 (dl, J = 8.59 Hz, 1 H) 7.23 (s, 1 H) 7.43 (d, J = 8.59 Hz, 2 H) 7.55 (d, J = 8.08 Hz, 2 H) 8.33 (s, 1 H)
[00462] [00462] LC / MS (LC-C method): Rt 2.90 min, MH + 619
[00463] [00463] [Į] D20: + 40.6 ° (c 0.32, DMF)
[00464] [00464] Chiral SFC (SFC-T method): Rt 2.48 min, without MH +, chiral purity 98.68%. Example 26A: synthesis of (1R *, 2R *) - 2 - ((3 - ((1- (4-chlorophenyl) -2- (5-fluoro-6- (trifluoromethoxy) indolin-1-yl) -2 -oxoethyl) amino) -5-methoxyphenoxy) methyl) -cyclopropanecarboxylic (Compound 26A)
[00465] [00465] A solution of 4-bromo-2-fluoro-1- (trifluoromethoxy) benzene [CAS 105529-58-6] (98.7 g, 381.1 mmol) in concentrated H2SO4 (98%, 200 mL) was cooled to 0 ° C with an ice bath. KNO3 (43.0 g, 425.3 mmol) was added in portions. After addition, the ice bath was removed and the mixture was stirred at room temperature for 16 h. The reaction mixture was poured into ice water (2 L) while stirring. The mixture was extracted with CH2Cl2 (3 x 500 ml). The combined organic layers were washed with a saturated aqueous solution of NaHCO3 (2 x 500 ml), brine (500 ml), dried over MgSO4, filtered and concentrated under reduced pressure to give 1-bromo-5-fluoro-2-nitro- 4- (trifluoromethoxy) benzene 26a (117.2 g), which was used in the next step without further purification. Synthesis of intermediate 26b:
[00466] [00466] To a stirred suspension of 1-bromo-5-fluoro-2-nitro-4- (trifluoromethoxy) benzene 26a (70.0 g, 230 mmol) and NH4Cl (123.2 g, 2.30 mol) in iPrOH (1 L) and water (330 mL) reducing iron powder (64.3 g, 1.15 mol) was added under N2 atmosphere. The reaction mixture was stirred at 60 ° C for 16 h. The reaction mixture was diluted with EtOAc (1 L) and filtered through Celite®. The filtrate was concentrated under reduced pressure. The residue was partitioned between EtOAc (1 L) and water (800 ml). The layers were separated and the organic phase was washed with brine
[00467] [00467] To a mixture of 2-bromo-4-fluoro-5- (trifluoromethoxy) aniline 26b (18.4 g, 67.2 mmol), ethinyl (trimethyl) silane (19.9 g, 202.4 mmol, 28.00 ml) in Et3N (300 ml) CuI (1.28 g, 6.72 mmol) and Pd (PPh3) 2Cl2 (2.40 g, 3.42 mmol) were added. The reaction mixture was heated under an N2 atmosphere at 90 ° C for 16 h. After cooling to room temperature, the mixture was diluted with MTBE (300 ml) and filtered through Celite®. The filtrate was concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel (ISCO®, 220 g SepaFlash® Silica Flash Column, eluent: 0 to 5% EtOAc gradient in petroleum ether @ 100 mL / min). 4-Fluoro-5- (trifluoromethoxy) -2 - ((trimethylsilyl) ethynyl) aniline 26c (16.1 g, 90% purity) was obtained as a brown oil. Synthesis of intermediate 26d:
[00468] [00468] A mixture of 4-fluoro-5- (trifluoromethoxy) -2- ((trimethylsilyl) ethynyl) aniline 26c (16.1 g, 55.3 mmol) and tBuOK (18.6 g, 165.8 mmol) in NMP (220.00 ml) it was heated to 90 ° C for 16 h under N2 atmosphere. After cooling to room temperature, the reaction mixture was poured into ice water (1 L) and extracted with MTBE (3 x 300 ml). The combined organic phases were washed with water (2 x 200 ml), brine (300 ml), dried over MgSO4, filtered and concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel (ISCO®, 120 g SepaFlash® Silica Flash Column, eluent: 0 to 5% EtOAc gradient in petroleum ether, flow rate = 85 mL / min) to give the product 5-fluoro-6- (trifluoromethoxy)) - 1H-indole 26d (11 g) as a dark green oil. The residue was combined with another fraction (total amount = 17.2 g) and further purified by distillation under reduced pressure (oil pump, eg 60 ~ 64 ° C) to provide 5-fluoro-6- (toxic trifluorome) -1H -indole 26d (14.7 g, 95% purity) as a colorless oil. Synthesis of intermediate 26e:
[00469] [00469] At 0 ° C, BH 3-Pyridine (1.2 mL, 11 mmol) was added slowly to a solution of 5-fluoro-6- (trifluoromethoxy) -1H-indole 26d (500 mg, 2.3 mmol ) in EtOH (3.2 ml). 6 N HCl (7.6 mL) was added slowly while maintaining the temperature below 10 ° C. The mixture was stirred at 0 ° C for 2 h. Water (100 m L) was added and the mixture was basified to pH 14 with concentrated NaOH (the temperature was kept below 20 ° C). CH 2Cl2 was added. The organic layer was separated, dried over MgSO4, filtered and the solvent was evaporated under reduced pressure to give 5-fluoro-6- (trifluoromethoxy) indoline 26e (550 mg). The compound was used in the next step without further purification. Synthesis of intermediate 26f:
[00470] [00470] To a mixture of 2-bromo-2- (4-chlorophenyl) acetic acid [CAS 3381-73-5] (0.61 g, 2.4 mmol), 5-fluoro-6- (trifluoromethoxy) indoline 26e (0.55 g, 2.2 mmol, 89% purity) and DMAP (0.027 g, 0.22 mmol) in CH2Cl2 (14 mL) EDCI (0.51 g, 2.7 mmol) was added. The mixture was stirred at room temperature for 18h. The mixture was diluted with a 10% K2CO3 solution in water. The layers were decanted. The organic layer was washed with water, dried over MgSO4, filtered and the solvent was concentrated under reduced pressure to give 2-bromo-2- (4-chlorophenyl) -1- (5-fluoro-6- (trifluoromethoxy) indolin-1 -il) ethanone 26f (1.1 g, purple oil). The compound was used in the next step without further purification. Synthesis of Compound 26A:
[00471] [00471] Compound 26A (135 mg) was synthesized from intermediate 26f using the procedures described for the synthesis of
[00472] [00472] H NMR (500 MHz, DMSO-d6) į ppm 0.87 - 0.94 (m, 1 H) 1.01 - 1.07 (m, 1 H) 1.55 (dq, J = 8 , 55, 4.40 Hz, 1 H) 1.60 - 1.68 (m, 1 H) 3.12 - 3.30 (m, 2 H) 3.61 (s, 3 H) 3.68 ( dd, J = 10.40, 7.57 Hz, 1 H) 3.85 (dd, J = 10.40, 6.31 Hz, 1 H) 4.01 - 4.08 (m, 1 H) 4 , 48 - 4.55 (m, 1 H) 5.57 (d, J = 8.83 Hz, 1 H) 5.75 (s, 1 H) 5.94 (ls, 1 H) 5.95 ( sl, 1 H) 6.46 (dl, J = 8.83 Hz, 1 H) 7.40 - 7.48 (m, 3 H) 7.54 (d, J = 8.51 Hz, 2 H) 8.16 (dl, J = 6.94 Hz, 1 H) 12.22 (ls, 1 H)
[00473] [00473] LC / MS (LC-C method): Rt 2.97 min, MH + 609
[00474] [00474] PF = 120 ° C Example 26B: synthesis of acid (1S *, 2S *) - 2 - ((3 - ((1- (4-chlorophenyl) -2- (5-fluoro-6- (trifluoromethoxy)) indolin-1-yl) -2-oxoethyl) amino) -5-methoxyphenoxy) methyl) -cyclopropanecarboxylic acid (Compound 26B) Synthesis of Compound 26B:
[00475] [00475] Compound 26B (150 mg) was synthesized from intermediate 26f using the procedure described for the synthesis of Compound 6B. Compound 26B: 1
[00476] [00476] H NMR (500 MHz, DMSO-d6) į ppm 0.86 - 0.94 (m, 1 H) 1.04 (dq, J = 8.20, 4.31 Hz, 1 H) 1, 55 (dq, J = 8.43, 4.33 Hz, 1 H) 1.58 - 1.67 (m, 1 H) 3.12 - 3.30 (m, 2 H) 3.61 (s, 3 H) 3.68 (dd, J = 10.40, 7.57 Hz, 1 H) 3.85 (dd, J = 10.25, 6.15 Hz, 1 H) 4.04 (q, J = 8.72 Hz, 1 H) 4.47 - 4.55 (m, 1 H) 5.57 (d, J = 8.83 Hz, 1 H) 5.75 (s, 1 H) 5.94 (sl, 1 H) 5.95 (sl, 1 H) 6.46 (dl, J = 8.51 Hz, 1 H) 7.40 - 7.48 (m, 3 H) 7.54 (d, J = 8.51 Hz, 2 H) 8.16 (dl, J = 6.94 Hz, 1 H) 12.21 (ls, 1 H)
[00477] [00477] LC / MS (LC-C method): Rt 2.97 min, MH + 609
[00478] [00478] PF = 126 ° C Example 27: synthesis of ((1s, 3s) -3- (3 - ((1- (4-chlorophenyl) -2- (5-fluoro- 6- (trifluoromethoxy) indolin- 1-yl) -2-oxoethyl) amino) -5-methoxyphenoxy) cyclobutene-carboxylic (Compound 27) and separation in Enantiomers 27A and 27B Synthesis of Compound 27 and chiral separation in Enantiomers 27A and 27B:
[00479] [00479] Compound 27 (175 mg) was synthesized from intermediate 26f using the procedure described for the synthesis of Compound 8. The two enantiomers were separated by chiral SFC (Stationary phase: Chiralcel® OD-H 5 µm 250 x 20 mm, mobile phase: 55% CO2, 45% EtOH) to give, after solidification from heptane / diisopropyl ether, the first eluted Enantiomer 27A (33 mg) and the second eluted Enantiomer 27B (35 mg). Compound 27: 1
[00480] [00480] H NMR (500 MHz, DMSO-d6) į ppm 2.01 - 2.17 (m, 2 H) 2.57 - 2.65 (m, 2 H) 2.66 - 2.77 (m , 1 H) 3.08 - 3.28 (m, 2 H) 3.61 (s, 3 H) 3.99 - 4.10 (m, 1 H) 4.43 - 4.57 (m, 2 H) 5.54 (d, J = 8.5 Hz, 1 H) 5.66 (s, 1 H) 5.86 (s, 1 H) 5.93 (s, 1 H) 6.49 (dl , J = 8.5 Hz, 1 H) 7.45 (d, J = 8.5 Hz, 3 H) 7.55 (d, J = 8.5 Hz, 2 H) 8.16 (dl, J = 6.9 Hz, 1 H) 12.06 - 12.47 (m, 1 H)
[00481] [00481] LC / MS (LC-C method): Rt 2.88 min, MH + 609
[00482] [00482] H NMR (500 MHz, DMSO-d6) į ppm 2.00 - 2.12 (m, 2 H) 2.57 - 2.65 (m, 3 H) 3.11 - 3.25 (m , 2 H) 3.60 (s, 3 H) 4.04 (dl, J = 7.3 Hz, 1 H) 4.40 - 4.48 (m, 1 H) 4.48 - 4.57 ( m, 1 H) 5.53 (dl, J = 8.5 Hz, 1 H) 5.65 (s, 1 H) 5.85 (s, 1 H) 5.92 (s, 1 H) 6, 48 (dl, J = 8.5 Hz, 1 H) 7.44 (dl, J = 8.5 Hz, 3 H) 7.54 (dl, J = 8.5 Hz, 2 H) 8.16 ( dl, J = 6.9 Hz, 1 H)
[00483] [00483] LC / MS (LC-D method): Rt 2.79 min, MH + 609
[00484] [00484] [Į] D20: -40.5 ° (c 0.252, DMF)
[00485] [00485] Chiral SFC (SFC-I method): Rt 1.18 min, without MH +, 100% chiral purity. 27B Enantiomer: 1
[00486] [00486] H NMR (500 MHz, DMSO-d6) į ppm 2.00 - 2.13 (m, 2 H) 2.54 - 2.67 (m, 3 H) 3.10 - 3.27 (m , 2 H) 3.60 (s, 3 H) 3.99 - 4.10 (m, 1 H) 4.40 - 4.48 (m, 1 H) 4.48 - 4.56 (m, 1 H) 5.54 (dl, J = 8.5 Hz, 1 H) 5.66 (s, 1 H) 5.86 (s, 1 H) 5.92 (s, 1 H) 6.48 (dl , J = 8.5 Hz, 1 H) 7.44 (dl, J = 8.5 Hz, 3 H) 7.54 (dl, J = 8.5 Hz, 2 H) 8.16 (dl, J = 6.9 Hz, 1 H)
[00487] [00487] LC / MS (LC-D method): Rt 2.79 min, MH + 609
[00488] [00488] [Į] D20: + 37.5 ° (c 0.333, DMF)
[00489] [00489] Chiral SFC (SFC-I method): Rt 2.56 min, without MH +, 100% chiral purity. Example 28: synthesis of (1R *, 2S *) - 2 - ((3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) methyl) -2-fluorocyclopropanecarboxylic acid (Compound 28A) and separation into Stereoisomers 28AA and 28AB and synthesis of (1S *, 2R *) - 2 - ((3 - ((1- (4-chlorophenyl ) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) methyl) -2-fluorocyclopropanecarboxylic acid (Compound 28B) and separation into Stereoisomers 28BA and 28BB
[00490] [00490] Rh2 (OPiv) 4.2H2O (2 mol%, 0.599 mmol, 387 mg) was added to a solution of 1 - (((2-fluoroalyl) oxy) methyl) -4-methoxybenzene [CAS 1673563-84- 2] (29.9 mmol) in anhydrous CH2Cl2 (86 mL) in a three-necked round-bottom flask equipped with a bubble chamber. After cooling the solution to 0 ° C, a commercially available 83% diazo ethyl acetate solution in dichloromethane (3 equiv., 89.85 mmol) in anhydrous CH2Cl2 (86 mL) was slowly added using a 24 mL flow rate micro-pump. /H. The mixture was stirred at room temperature until completion of the reaction (indicated by TLC and F NMR analysis) and concentrated under reduced pressure. The crude residue was purified by column chromatography on silica gel (petroleum ether / EtOAc, 9/1 to 7/3) to give a mixture of diastereomers in 60% yield (dr 53:47). The diastereomers were separated by column chromatography on silica gel (petroleum ether / EtOAc, 100/0 to 80/20) to give 2-fluoro-2 - ((((4-methoxybenzyl) oxy) methyl) trans-cyclopropanecarboxylate -ethyl 28a and 2-fluoro-2 - cis-ethyl cyclopropanecarboxylate ((((4-methoxybenzyl) oxy) methyl) 28b. Synthesis of intermediate 28c:
[00491] [00491] DDQ (1.5 equiv., 27.9 mmol, 6.33 g) was added to a solution of trans-ethyl 2-fluoro-2 - (((4-methoxybenzyl) oxy) methyl) cyclopropanecarboxylate 28a (1 equiv., 18.6 mmol, 5.25 g) in dichloromethane (340 mL) and water (30 mL) at 0 ° C. The mixture was stirred for 20 h. A solution of saturated aqueous NaHCO 3 was added and the mixture was stirred for 30 minutes. The aqueous layer was extracted three times with CH2Cl2. The combined organic layers were washed with a saturated solution of NaHCO3 and brine. The organic layer was evaporated under reduced pressure and the crude residue was purified by chromatography on silica gel (petroleum ether / EtOAc, 9/1 to 6/4) to give 2-fluoro-2- (hydroxymethyl) -cyclopropanecarboxylate trans-ethyl 28c (876 mg). Synthesis of intermediate 28d:
[00492] [00492] DDQ (1.5 equiv., 15.8 mmol, 3.6 g) was added to a solution of cis-ethyl 2-fluoro-2 - (((4-methoxybenzyl) oxy) methyl) cyclopropanecarboxylate 28b (1 equiv., 10.6 mmol, 2.98 g) in dichloromethane (193 ml) and water (17 ml) at 0 ° C. The mixture was stirred for 20 h. A saturated aqueous NaHCO3 solution was added and the mixture was stirred for 30 minutes. The aqueous layer was extracted three times with CH2Cl2. The combined organic layers were washed with a saturated solution of
[00493] [00493] Under a flow of N2 at 10 ºC, di-tert-butyl azodicarboxylate (948 mg, 4,118 mmol) was added portion by portion to a solution of 3-methoxy-5-nitrophenol [7145-49-5] ( 633 mg, 3.743 mmol), trans-ethyl 2-fluoro-2- (hydroxymethyl) cyclopropanecarboxylate 28c (607 mg, 3.743 mmol) and PPh3 (1.08 g, 4.118 mmol) in THF (30 mL). The reaction was stirred at room temperature under N2 for 18 h. The solution was concentrated under reduced pressure. The crude residue was purified by column chromatography on silica gel (15-40 µm, 80 g, heptane / EtOAc 95/5 to 80/20). The pure fractions were combined and concentrated under reduced pressure to give trans-methyl 2-fluoro-2 - ((3-methoxy-5-nitrophenoxy) methyl) cyclopropane-carboxylate 28e (930 mg). Synthesis of intermediate 28f:
[00494] [00494] A solution of trans-methyl 2-fluoro-2 - ((3-methoxy-5-nitrophenoxy) methyl) cyclopropane-carboxylate 28e (810 mg, 2.586 mmol) in EtOH (20 mL) and THF (10 mL ) containing a catalytic amount of 10% Pd / C (275 mg, 0.259 mmol) was hydrogenated under atmospheric pressure of H2 at room temperature for 4 h. The catalyst was removed by filtration over a short pad of Celite® and the filter cake was rinsed several times with EtOH. The combined filtrates were evaporated under reduced pressure to give trans-methyl 2 - ((3-amino-5-methoxyphenoxy) -methyl) -2-fluorocyclopropanecarboxylate 28f (710 mg) which was used in the next step without further purification. Synthesis of intermediate 28g and separation in Stereoisomers 28h, 28i, 28j and 28k:
[00495] [00495] A mixture of 2-bromo-2- (4-chlorophenyl) -1- (6- (trifluoromethoxy) indolin-1-yl) ethanone 1c (927 mg, 2.133 mmol), 2 - ((3- amino- Transmethyl 5-methoxyphenoxy) -methyl) -2-fluorocyclopropanecarboxylate (725 mg, 2.559 mmol) and diisopropylethylamine (735 µL, 4.265 mmol) in CH3CN (4 mL) was stirred at 80 ° C for 12 h. The solution was concentrated under reduced pressure. The residue was taken up with EtOAc. The organic layer was washed with 1 N HCl, water, dried over MgSO4, filtered and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (15-40 µm, 40 g, heptane / EtOAc 95/5 to 80/20). The pure fractions were combined and evaporated under reduced pressure to give 2 - ((3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethyl) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) methyl) - methyl trans-2-fluorocyclopropanecarboxylate 28g (550 mg). The four stereoisomers were separated by chiral SFC (Stationary phase: Chiralcel® OD-H 5 µm 250 x 20 mm, mobile phase: 70% CO2, 30% EtOH) to give 28h (118 mg), 28i (114 mg ), 28j (158 mg) and 28k (165 mg). Synthesis of Compound 28A and separation in Stereoisomers 28AA and 28AB:
[00496] [00496] LiOH monohydrate (23.3 mg, 0.556 mmol) was added to a solution of the stereoisomer 28h (118 mg, 0.185 mmol) in THF / MeOH / water (1/1/1) (2 mL). The reaction mixture was stirred at room temperature for 4 h. The mixture was diluted with water and ice, slowly acidified with 1 N HCl and extracted with EtOAc. The combined organic layers were washed with water, dried over MgSO4, filtered and concentrated under reduced pressure to give, after crystallization from heptane / diisopropyl ether, (1R *, 2S *) - 2- ((3- ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) - 5-methoxyphenoxy) methyl) -2-fluorocyclopropanecarboxylic 28A (110 mg) (during this reaction occurred total racemization in the central chiral center).
[00497] [00497] A second batch of Compound 28A (100 mg) was similarly obtained starting from stereoisomer 28i. The two batches were combined. The two stereoisomers were separated by chiral SFC (Stationary phase: Chiralcel® OD-H 5 µm 250 x 20 mm, mobile phase: 65% CO2, 35% EtOH) to give the first eluted stereoisomer (94 mg) and the stereoisomer eluted second (80 mg). The stereoisomer eluted first was solidified from diisopropyl ether to give Stereoisomer 28AA (47 mg). The stereoisomer eluted second was solidified from heptane to give Stereoisomer 28AB (37 mg). Synthesis of Compound 28B and separation in Stereoisomers 28BA and 28BB:
[00498] [00498] LiOH monohydrate (31.2 mg, 0.744 mmol) was added to a solution of the 28j stereoisomer (158 mg, 0.248 mmol) in THF / MeOH / water (1/1/1) (2 mL). The reaction mixture was stirred at room temperature for 4 h. The mixture was diluted with water and ice, slowly acidified with 1 N HCl and extracted with EtOAc. The combined organic layers were washed with water, dried over MgSO4, filtered and concentrated under reduced pressure to give, after crystallization from MeOH / water, acid (1S *, 2R *) - 2 - ((3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) methyl) -2-fluorocyclopropane-carboxylic 28B (100 mg) (during this reaction total racemization occurred at the central chiral center).
[00499] [00499] A second batch of 28A (105 mg) was similarly obtained starting from the stereoisomer 28k.
[00500] [00500] The two stereoisomers were separated by chiral SFC (Stationary phase: Chiralcel® OD-H 5 µm 250 x 30 mm, mobile phase: 60% CO2, 30% MeOH) to give the eluted stereoisomer first ( 88 mg) and the second eluted stereoisomer (78 mg). The stereoisomer eluted first was solidified from heptane / diisopropyl ether to give Stereoisomer 28BA (54 mg). The second eluted stereoisomer was solidified from heptane / diisopropyl ether to give Stereoisomer 28BB (60 mg). Stereoisomer 28AA: 1
[00501] [00501] H NMR (500 MHz, DMSO-d6) į ppm 1.37 (dt, J = 11.7, 7.1 Hz, 1 H) 1.62 - 1.76 (m, 1 H) 2, 18 - 2.31 (m, 1 H) 3.07 - 3.23 (m, 2 H) 3.62 (s, 3 H) 3.98 - 4.10 (m, 1 H) 4.15 - 4.29 (m, 1 H) 4.43 (dd, J = 18.9, 12.0 Hz, 1 H) 4.48 - 4.57 (m, 1 H) 5.58 (d, J = 8.8 Hz, 1 H) 5.78 (s, 1 H) 5.97 (s, 1 H) 5.99 (s, 1 H) 6.49 (dl, J = 8.8 Hz, 1 H ) 7.01 (dl, J = 7.9 Hz, 1 H) 7.33 (d, J = 8.2 Hz, 1 H) 7.44 (d, J = 8.5 Hz, 2 H) 7 , 55 (d, J = 8.2 Hz, 2 H) 8.03 (s, 1 H) 12.71 (sl, 1 H)
[00502] [00502] LC / MS (LC-C method): Rt 2.83 min, MH + 609
[00503] [00503] [Į] D20: -43.3 ° (c 0.3, DMF)
[00504] [00504] Chiral SFC (SFC-F method): Rt 1.98 min, without MH +, 100% chiral purity. Stereoisomer 28AB: 1
[00505] [00505] H NMR (500 MHz, DMSO-d6) į ppm 1.32 - 1.42 (m, 1 H) 1.62 - 1.75 (m, 1 H) 2.24 (dt, J = 18 , 5, 9.2 Hz, 1 H) 3.08 - 3.25 (m, 2 H) 3.62 (s, 3 H) 3.99 - 4.08 (m, 1 H) 4.13 - 4.26 (m, 1 H) 4.39 - 4.57 (m, 2 H) 5.58 (dl, J = 8.8 Hz, 1 H) 5.78 (s, 1 H) 5.96 (sl, 1 H) 5.99 (sl, 1 H) 6.49 (dl, J = 8.8 Hz, 1 H) 7.01 (dl, J = 7.6 Hz, 1 H) 7.33 (dl, J = 8.2 Hz, 1 H) 7.44 (dl, J = 8.2 Hz, 2 H) 7.55 (dl, J = 8.2 Hz, 2 H) 8.03 (sl , 1 H) 12.71 (ls, 1 H)
[00506] [00506] LC / MS (LC-C method): Rt 2.84 min, MH + 609
[00507] [00507] [Į] D20: + 52.5 ° (c 0.301, DMF)
[00508] [00508] Chiral SFC (SFC-F method): Rt 3.29 min, without MH +, 100% chiral purity. Stereoisomer 28BA: 1
[00509] [00509] H NMR (500 MHz, DMSO-d6) į ppm 1.32 - 1.42 (m, 1 H)
[00510] [00510] LC / MS (LC-D method): Rt 2.71 min, MH + 609
[00511] [00511] [Į] D20: -57.1 ° (c 0.31, DMF)
[00512] [00512] Chiral SFC (SFC-F method): Rt 2.26 min, without MH +, 100% chiral purity. Stereoisomer 28BB: 1
[00513] [00513] H NMR (500 MHz, DMSO-d6) į ppm 1.35 (dt, J = 11.7, 7.1 Hz, 1 H) 1.59 - 1.71 (m, 1 H) 2, 17 - 2.28 (m, 1 H) 3.08 - 3.25 (m, 2 H) 3.62 (s, 3 H) 4.04 (td, J = 10.5, 7.1 Hz, 1 H) 4.17 - 4.29 (m, 1 H) 4.43 (dd, J = 19.5, 11.7 Hz, 1 H) 4.52 (td, J = 10.4, 6, 3 Hz, 1 H) 5.59 (d, J = 9.1 Hz, 1 H) 5.78 (t, J = 1.9 Hz, 1 H) 5.96 (s, 1 H) 5.99 (s, 1 H) 6.49 (d, J = 8.8 Hz, 1 H) 7.01 (dd, J = 8.0, 1.4 Hz, 1 H) 7.33 (d, J = 8.2 Hz, 1 H) 7.44 (d, J = 8.5 Hz, 2 H) 7.55 (d, J = 8.2 Hz, 2 H) 8.03 (s, 1 H) 12 79 (ls, 1 H)
[00514] [00514] LC / MS (LC-D method): Rt 2.70 min, MH + 609
[00515] [00515] [Į] D20: + 38.1 ° (c 0.289, DMF)
[00516] [00516] Chiral SFC (SFC-F method): Rt 3.68 min, without MH +, 100% chiral purity. Example 29: synthesis of (1S *, 2S *) - 2 - ((3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) methyl) -2-fluorocyclopropanecarboxylic acid (Compound 29A) and separation in Stereoisomers 29AA and 29AB and synthesis of (1R *, 2R *) - 2 - ((3 - ((1- (4-chlorophenyl) ) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) methyl) -2-fluorocyclopropanecarboxylic acid (Compound 29B) and separation in Stereoisomers 29BA and 29BB
[00517] [00517] Under a flow of N2 at 10ºC, di-tert-butyl azodicarboxylate (750 mg, 3.256 mmol) was added portion by portion to a solution of 3-methoxy-5-nitrophenol [7145-49-5] (501 mg, 2.96 mmol), cis-ethyl 2-fluoro-2- (hydroxymethyl) cyclopropanecarboxylate 28d (480 mg, 2.96 mmol) and PPh3 (854 mg, 3.256 mmol) in THF (23 mL). The reaction was stirred at room temperature under N2 for 18 h. The solution was concentrated under reduced pressure. The crude residue was purified by column chromatography on silica gel (15-40 µm, 80 g, heptane / EtOAc 70/30). The pure fractions were combined and concentrated under reduced pressure to give cis-methyl 2-fluoro-2 - ((3-methoxy-5-nitrophenoxy) methyl) cyclopropanecarboxylate 29a (660 mg). Synthesis of intermediate 29b:
[00518] [00518] A solution of cis-methyl 2-fluoro-2 - ((3-methoxy-5-nitrophenoxy) methyl) cyclopropane-carboxylate 29a (610 mg, 1.947 mmol) in EtOH (15 mL) and THF (7, 5 ml) containing a catalytic amount of 10% Pd / C (207 mg, 0.195 mmol) was hydrogenated under atmospheric pressure of H2 at room temperature for 4 h. The catalyst was removed by filtration over a short pad of Celite® and the filter cake was rinsed several times with EtOH. The combined filtrates were evaporated to give cis-methyl 2 - ((3-amino-5-methoxyphenoxy) methyl) -2-fluorocyclopropane-carboxylate 29b (560 mg) which was used in the next step without further purification. Synthesis of intermediate 29c and separation in Stereoisomers 29d, 29e, 29f and 29g:
[00519] [00519] A mixture of 2-bromo-2- (4-chlorophenyl) -1- (6- (trifluoromethoxy) indolin-1-yl) ethanone 1c (716 mg, 1.647 mmol), 2 - ((3- amino- Cis-methyl 5-methoxy-phenoxy) -methyl) -2-fluorocyclopropanecarboxylate 29b (560 mg, 1.977 mmol) and diisopropylethylamine (568 µL, 3.295 mmol) in CH3CN (3.5 mL) was stirred at 80 ° C for 12 h. The solvent was concentrated under reduced pressure. The residue was taken up with EtOAc. The organic layer was washed with 1 N HCl, water, dried over MgSO4, filtered and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (15-40 µm, 40 g, heptane / EtOAc 95/5 to 80/20). The pure fractions were combined and evaporated under reduced pressure to give 2 - ((3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethyl) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) methyl) -cis- 2-fluorocyclopropanecarboxylate 29c (500 mg). The four stereoisomers were separated by chiral SFC (Stationary phase: Chiralpak® AD-H 5 µm 250 x 30 mm, mobile phase: 65% CO2, 35% EtOH) to give a mixture of 29d + 29e (250 mg) , 29f (125 mg) and 29g (114 mg). The 29d + 29e mixture was further separated by chiral SFC (Stationary phase: Chiralpak® AD-H 5 µm 250 x 30 mm, mobile phase: 75% CO2, 25% EtOH) to give 29d (88 mg) and 29e (66 mg).
[00520] [00520] LiOH monohydrate (17.4 mg, 0.414 mmol) was added to a solution of stereoisomer 29d (88 mg, 0.138 mmol) in THF / MeOH / water (1/1/1) (1 mL). The reaction mixture was stirred at room temperature for 4 h. The mixture was diluted with water and ice, slowly acidified with 1 N HCl and extracted with EtOAc. The combined organic layers were washed with water, dried over MgSO4, filtered and concentrated under reduced pressure to give acid (1S *, 2S *) - 2 - ((3 - ((1- (4-chlorophenyl) -2-oxo- 2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) methyl) -2-fluorocyclopropanecarboxylic 29A (80 mg) (during this reaction total racemization occurred at the central chiral center).
[00521] [00521] A second batch of 29A (90 mg) was similarly obtained starting from the stereoisomer 29f. The two batches were combined. The two stereoisomers were separated by chiral SFC (Stationary phase: Chiralcel® OD-H 5 µm 250 x 20 mm, mobile phase: 65% CO2, 35% MeOH) and further purified by flash chromatography on silica gel (15 -40 µm, 4 g, CH2Cl2 / MeOH 99/1) to give the stereoisomer eluted first (43 mg) and the stereoisomer eluted second (40 mg). The stereoisomer eluted first was solidified in heptane / diisopropyl ether to give stereoisomer 29AA (29 mg). The stereoisomer eluted second was solidified in heptane / diisopropyl ether to give stereoisomer 29AB (27 mg). Synthesis of Compound 29B and separation in Stereoisomers 29BA and 29BB:
[00522] [00522] LiOH monohydrate (13 mg, 0.311 mmol) was added to a solution of stereoisomer 29e (66 mg, 0.104 mmol) in THF / MeOH / water (1/1/1) (1 mL). The reaction mixture was stirred at room temperature for 4 h. The mixture was diluted with water and ice, slowly acidified with 1 N HCl and extracted with EtOAc. The combined organic layers were washed with water, dried over MgSO4, filtered and concentrated under reduced pressure to give, after crystallization from MeOH / water, acid (1R *, 2R *) - 2 - ((3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) methyl) -2-fluorocyclopropane-carboxylic 29B (60 mg) (during this reaction total racemization occurred at the central chiral center).
[00523] [00523] A second batch of 29A (100 mg) was similarly obtained starting from the stereoisomer 29g. The two batches were combined. The two stereoisomers were separated by chiral SFC (Stationary phase: Chiralcel® OD-H 5 µm 250 x 20 mm, mobile phase: 70% CO2, 30% MeOH) and further purified by flash chromatography on silica gel (15 -40 µm, 4 g, CH2Cl2 / MeOH 99/1) to give the stereoisomer eluted first (38 mg) and the stereoisomer eluted second (31 mg). The stereoisomer eluted first was solidified in heptane / diisopropyl ether to give stereoisomer 29BA (24 mg). The second eluted stereoisomer was solidified in heptane / diisopropyl ether to give stereoisomer 29BB (20 mg). Stereoisomer 29AA: 1
[00524] [00524] H NMR (500 MHz, DMSO-d6) į ppm 1.30 - 1.37 (m, 1 H) 1.69 (dt, J = 19.7, 6.9 Hz, 1 H) 1, 98 - 2.06 (m, 1 H) 3.08 - 3.24 (m, 2 H) 3.63 (s, 3 H) 4.01 - 4.09 (m, 1 H) 4.10 - 4.23 (m, 2 H) 4.48 - 4.57 (m, 1 H) 5.59 (d, J = 8.8 Hz, 1 H) 5.81 (s, 1 H) 5.99 (dl, J = 5.7 Hz, 2 H) 6.49 (dl, J = 8.8 Hz, 1 H) 7.01 (dl, J = 7.9 Hz, 1 H) 7.33 (d , J = 7.9 Hz, 1 H) 7.44 (d, J = 8.5 Hz, 2 H) 7.55 (d, J = 8.2 Hz, 2 H) 8.03 (sl, 1 H) 12.58 (ls, 1 H)
[00525] [00525] LC / MS (LC-D method): Rt 2.67 min, MH + 609
[00526] [00526] [Į] D20: -15.7 ° (c 0.242, DMF)
[00527] [00527] Chiral SFC (SFC-P method): Rt 2.53 min, without MH +, 100% chiral purity. Stereoisomer 29AB: 1
[00528] [00528] H NMR (500 MHz, DMSO-d6) į ppm 1.29 - 1.38 (m, 1 H) 1.69 (dt, J = 19.5, 6.5 Hz, 1 H) 1, 97 - 2.10 (m, 1 H) 3.08 - 3.25 (m, 2 H) 3.63 (s, 3 H) 4.00 - 4.10 (m, 1 H) 4.10 - 4.23 (m, 2 H) 4.52 (dl, J = 6.0 Hz, 1 H) 5.60 (dl, J = 8.8 Hz, 1 H) 5.82 (sl, 1 H) 6.00 (dl, J = 6.6 Hz, 2 H) 6.50 (dl, J = 8.5 Hz, 1 H) 7.01 (dl, J = 7.6 Hz, 1 H) 7, 34 (dl, J = 7.9 Hz, 1 H) 7.44 (dl, J = 8.2 Hz, 2 H) 7.55 (dl, J = 8.2 Hz, 2 H) 8.04 ( ls, 1 H) 12.58 (ls, 1 H)
[00529] [00529] LC / MS (LC-D method): Rt 2.67 min, MH + 609
[00530] [00530] [Į] D20: + 77.4 ° (c 0.323, DMF)
[00531] [00531] Chiral SFC (SFC-P method): Rt 4.47 min, without MH +, chiral purity 99.20%. Stereoisomer 29BA: 1
[00532] [00532] H NMR (500 MHz, DMSO-d6) į ppm 1.33 (td, J = 9.8, 6.6 Hz, 1 H) 1.69 (dt, J = 19.9, 6.9 Hz, 1 H) 2.03 (ddd, J = 9.5, 7.1, 3.0 Hz, 1 H) 3.08 - 3.25 (m, 2 H) 3.63 (s, 3 H ) 4.05 (td, J = 10.3, 7.1 Hz, 1 H) 4.10 - 4.23 (m, 2 H) 4.52 (td, J = 10.2, 6.3 Hz , 1 H) 5.59 (d, J = 8.8 Hz, 1 H) 5.80 - 5.84 (m, 1 H) 6.00 (dl, J = 7.6 Hz, 2 H) 6 , 49 (dl, J = 8.8 Hz, 1 H) 7.01 (dd, J = 8.0, 1.4 Hz, 1 H) 7.33 (d, J = 8.2 Hz, 1 H ) 7.44 (d, J = 8.5 Hz, 2 H) 7.55 (d, J = 8.2 Hz, 2 H) 8.03 (s, 1 H) 12.58 (sl, 1 H )
[00533] [00533] LC / MS (LC-D method): Rt 2.67 min, MH + 609
[00534] [00534] [Į] D20: -74.2 ° (c 0.302, DMF)
[00535] [00535] Chiral SFC (SFC-P method): Rt 2.37 min, without MH +, 100% chiral purity. Stereoisomer 29BB: 1
[00536] [00536] H NMR (500 MHz, DMSO-d6) į ppm 1.34 (td, J = 9.8, 6.6 Hz, 1 H) 1.69 (dt, J = 19.9, 6.9 Hz, 1 H) 2.02 (ddd, J = 9.5, 7.1, 3.0 Hz, 1 H) 3.09 - 3.25 (m, 2 H) 3.63 (s, 3 H ) 4.05 (td, J = 10.4, 7.3 Hz, 1 H) 4.10 - 4.22 (m, 2 H) 4.52 (td, J = 10.3, 6.5 Hz , 1 H) 5.59 (d, J = 8.8 Hz, 1 H) 5.81
[00537] [00537] LC / MS (LC-D method): Rt 2.70 min, MH + 609
[00538] [00538] [Į] D20: + 12.0 ° (c 0.3, DMF)
[00539] [00539] Chiral SFC (SFC-P method): Rt 3.73 min, without MH +, chiral purity 99.14%. Example 30: synthesis of (1s, 3s) -3 - ((3 - ((1- (4-chlorophenyl) -2- (5-fluoro-6- (trifluoromethoxy) indolin-1-yl) -2-oxoethyl acid ) amino) -5-methoxyphenoxy) methyl) - cyclobutanecarboxylic (Compound 30) and chiral separation in Enantiomers 30A and 30B Synthesis of Compound 30 and chiral separation in Enantiomers 30A and 30B:
[00540] [00540] Compound 30 (105 mg) was synthesized from intermediate 26f using the procedures described for the synthesis of Compound 10. The two enantiomers were separated by chiral SFC (Stationary phase: Chiralcel® OJ-H 5 µm 250 x 20 mm, mobile phase: 70% CO2, 30% MeOH) to give, after lyophilization in CH3CN / water, the Enantiomer eluted first 30A (43 mg) and the Enantiomer eluted second 30B (47 mg). 30A Enantiomer: 1
[00541] [00541] H NMR (500 MHz, DMSO-d6) į ppm 1.85 - 1.98 (m, 2 H) 2.17 - 2.28 (m, 2 H) 2.53 - 2.59 (m , 1 H) 2.96 (quin, J = 8.9 Hz, 1 H) 3.09
[00542] [00542] LC / MS (LC-C method): Rt 3.08 min, MH + 623
[00543] [00543] [Į] D20: + 42.6 ° (c 0.298, DMF)
[00544] [00544] Chiral SFC (SFC-F method): Rt 2.91 min, without MH +, 100% chiral purity. 30B Enantiomer: 1
[00545] [00545] H NMR (500 MHz, DMSO-d6) į ppm 1.87 - 1.99 (m, 2 H) 2.16 - 2.29 (m, 2 H) 2.53 - 2.59 (m , 1 H) 2.96 (tl, J = 8.8 Hz, 1 H) 3.11 - 3.23 (m, 2 H) 3.61 (s, 3 H) 3.77 (dl, J = 6.0 Hz, 2 H) 4.05 (td, J = 10.2, 7.3 Hz, 1 H) 4.51 (td, J = 10.3, 6.5 Hz, 1 H) 5, 56 (d, J = 8.8 Hz, 1 H) 5.74 (s, 1 H) 5.93 (s, 2 H) 6.45 (dl, J = 8.8 Hz, 1 H) 7, 44 (dl, J = 8.2 Hz, 3 H) 7.54 (d, J = 8.5 Hz, 2 H) 8.16 (dl, J = 6.9 Hz, 1 H) 11.43 - 12.72 (m, 1 H)
[00546] [00546] LC / MS (LC-C method): Rt 3.07 min, MH + 623
[00547] [00547] [Į] D20: -44.2 ° (0.217 c, DMF)
[00548] [00548] Chiral SFC (SFC-F method): Rt 4.10 min, without MH +, chiral purity 99.09%. Example 31: synthesis of (1r, 3r) -3 - ((3 - ((1- (4-chlorophenyl) -2- (5-fluoro-6- (trifluoromethoxy) indolin-1-yl) -2-oxoethyl acid ) amino) -5-methoxyphenoxy) methyl) - cyclobutanecarboxylic (Compound 31) and chiral separation in Enantiomers 31A and 31B
[00549] [00549] Compound 31 (75 mg) was synthesized from intermediate 26f using the procedure described for the synthesis of Compound 11. The two enantiomers were separated by chiral SFC (Stationary phase: Chiralcel® OJ-H 5 µm 250 x 20 mm, mobile phase: 70% CO2, 30% MeOH) to give, after lyophilization in CH3CN / water, the Enantiomer eluted first 31A (23 mg) and the Enantiomer eluted second 31B (24 mg). 31A Enantiomer: 1
[00550] [00550] H NMR (500 MHz, DMSO-d6) į ppm 1.90 - 2.03 (m, 2 H) 2.20 - 2.30 (m, 2 H) 2.55 - 2.62 (m , 1 H) 3.07 (tl, J = 7.7 Hz, 1 H) 3.12 - 3.24 (m, 2 H) 3.62 (s, 3 H) 3.87 (dl, J = 6.9 Hz, 2 H) 4.05 (td, J = 10.3, 7.1 Hz, 1 H) 4.51 (td, J = 10.4, 6.6 Hz, 1 H) 5, 57 (d, J = 8.8 Hz, 1 H) 5.78 (s, 1 H) 5.94 (s, 1 H) 5.96 (s, 1 H) 6.45 (d, J = 8 , 8 Hz, 1 H) 7.44 (d, J = 8.5 Hz, 3 H) 7.54 (d, J = 8.5 Hz, 2 H) 8.16 (dl, J = 6.9 Hz, 1 H)
[00551] [00551] LC / MS (LC-C method): Rt 3.07 min, MH + 623
[00552] [00552] [Į] D20: + 43.1 ° (c 0.255, DMF)
[00553] [00553] Chiral SFC (SFC-F method): Rt 3.25 min, without MH +, 100% chiral purity. 31B Enantiomer: 1
[00554] [00554] H NMR (500 MHz, DMSO-d6) į ppm 1.90 - 2.03 (m, 2 H)
[00555] [00555] LC / MS (LC-C method): Rt 3.07 min, MH + 623
[00556] [00556] [Į] D20: -43.4 ° (c 0.244, DMF)
[00557] [00557] Chiral SFC (SFC-F method): Rt 4.85 min, without MH +, chiral purity 99.09%. Example 32: synthesis of (1s, 3s) -3- (3 - ((2- (6-chloro-5-methoxy-indolin-1-yl) -1- (4-chlorophenyl) -2-oxoethyl) amino acid ) -5- methoxyphenoxy) cyclobutanecarboxylic (Compound 32) and chiral separation in Enantiomers 32A and 32B.
[00558] [00558] Intermediate 32a (3.58 g) was synthesized from 6-chloro-5-methoxy-indoline [CAS 1369041-89-3] using the procedure described for the synthesis of intermediate 26f. Synthesis of intermediate 32b:
[00559] [00559] Intermediate 32b (210 mg) was synthesized from intermediate 32a using the procedure described for the synthesis of intermediate 8d. Synthesis of Compound 32 and chiral separation in Enantiomers 32A and 32B:
[00560] [00560] Compound 32 (165 mg) was synthesized from intermediate 32b using the procedure described for the synthesis of Compound 28. The two enantiomers were separated by chiral SFC (Stationary phase: Chiralcel® OD-H 5 µm 250 x 30 mm, mobile phase: 50% CO2, 50% EtOH) to give, after purification by flash chromatography on silica gel (15-40 µm; 4 g, CH2Cl2 / CH3OH 97/3) and solidification from heptane, diisopropyl ether, the Enantiomer eluted first 32A (26 mg) and the Enantiomer eluted second 32B (31 mg). Compound 32: 1
[00561] [00561] H NMR (500 MHz, DMSO-d6) į ppm 2.03 - 2.15 (m, 2 H) 2.57 - 2.66 (m, 2 H) 2.66 - 2.75 (m , 1 H) 3.06 - 3.23 (m, 2 H) 3.61 (s, 3 H) 3.80 (s, 3 H) 3.95 (td, J = 10.4, 7.3 Hz, 1 H) 4.43 - 4.52 (m, 2 H) 5.50 (d, J = 8.5 Hz, 1 H) 5.65 (t, J = 1.9 Hz, 1 H) 5.86 (s, 1 H) 5.93 (s, 1 H) 6.40 (d, J = 8.5 Hz, 1 H) 7.10 (s, 1 H) 7.44 (d, J = 8.5 Hz, 2 H) 7.54 (d, J = 8.5 Hz, 2 H) 8.11 (s, 1 H) 12.26 (sl, 1 H)
[00562] [00562] LC / MS (LC-C method): Rt 2.66 min, MH + 569 32A enantiomer: 1
[00563] [00563] H NMR (500 MHz, DMSO-d6) į ppm 2.03 - 2.15 (m, 2 H) 2.57 - 2.66 (m, 2 H) 2.66 - 2.76 (m , 1 H) 3.04 - 3.24 (m, 2 H) 3.57 - 3.64 (m, 3 H) 3.80 (s, 3 H) 3.89 - 4.00 (m, 1 H) 4.42 - 4.54 (m, 2 H) 5.50 (dl, J = 8.8 Hz, 1 H) 5.65 (s, 1 H) 5.86 (s, 1 H) 5 , 93 (s, 1 H) 6.40 (dl, J = 8.5 Hz, 1 H) 7.10 (s, 1 H) 7.44 (d, J = 8.2 Hz, 2 H) 7 , 54 (dl, J = 8.2 Hz, 2 H) 8.11 (s, 1 H) 12.26 (ls, 1 H)
[00564] [00564] LC / MS (LC-D method): Rt 2.56 min, MH + 569
[00565] [00565] [Į] D20: -55.4 ° (c 0.332, DMF)
[00566] [00566] Chiral SFC (SFC-R method): Rt 1.73 min, without MH +, 100% chiral purity. 32B Enantiomer: 1
[00567] [00567] H NMR (500 MHz, DMSO-d6) į ppm 2.02 - 2.14 (m, 2 H)
[00568] [00568] LC / MS (LC-D method): Rt 2.56 min, MH + 569
[00569] [00569] [Į] D20: + 53.4 ° (c 0.35, DMF)
[00570] [00570] Chiral SFC (SFC-R method): Rt 3.16 min, without MH +, chiral purity 99.59%. Example 33: synthesis of 3 - ((3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenoxy) amino acid ) - cyclobutanecarboxylic (Compound 33) and chiral separation in Stereoisomers 33A, 33B, 33C and 33D.
[00571] [00571] Under nitrogen, a mixture of 3-methoxy-5-nitroaniline [CAS 586-10-7] (0.50 g, 2.973 mmol), ethyl 3-oxocyclobutanecarboxylate [CAS 87121-89-9] (1, 27 g, 8.92 mmol) and acetic acid (0.34 mL, 5.947 mmol) in dry EtOH (26 mL) was stirred at room temperature for 30 min. NaBH3CN (0.374 g, 5.947 mmol) was added and the mixture was stirred at room temperature for 18 h. Brine was added and the mixture was extracted twice with CH2Cl2. The combined organic layers were dried over MgSO4, filtered and concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel (30 µm, 24 g, heptane / EtOAc from 85/15 to 75/25). The pure fractions were combined and concentrated under reduced pressure to give ethyl 3 - ((3-methoxy-5-nitrophenyl) amino) cyclobutane-carboxylate 33a (820 mg). The compound was used in the next step without further purification. Synthesis of intermediate 33b:
[00572] [00572] A solution of ethyl 3 - ((3-methoxy-5-nitrophenyl) amino) cyclobutane-carboxylate 33a (820 mg, 2.8 mmol) in EtOH (16 mL) and EtOAc (14 mL) containing an amount catalytic 10% Pd / C (300 mg, 0.28 mmol) was hydrogenated under atmospheric pressure of H2 at room temperature for 18 h. The catalyst was removed by filtration over a short pad of Celite® and the filter cake was rinsed several times with EtOAc. The combined filtrates were evaporated under reduced pressure to give ethyl 3 - ((3-amino-5-methoxyphenyl) amino) cyclobutanecarboxylate 33b (800 mg) which was used in the next step without further purification. Synthesis of intermediate 33c:
[00573] [00573] A mixture of 2-bromo-2- (4-chlorophenyl) -1- (6- (trifluoromethoxy) indolin-1-yl) ethanone 1c (0.936 g, 2.154 mmol), 3 - ((3- amino- Ethyl 5-methoxyphenyl) amino) cyclobutanecarboxylate 33b (0.74 g, 2.8 mmol) and diisopropylethylamine (0.742 mL, 4.307 mmol) in CH3CN (11 mL) was stirred at 80 ° C for 18 h. The mixture was concentrated under reduced pressure. Purification by flash chromatography on silica gel (30 µm, 40 g, heptane / EtOAc from 85/15 to 70/30) was performed. The pure fractions were combined and evaporated to dryness to give 3 - ((3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) Ethyl methoxyphenyl) amino) cyclobutanecarboxylate 33c (600 mg). Synthesis of Compound 33 and chiral separation in Stereoisomers 33A, 33B, 33C and 33D:
[00574] [00574] A solution of LiOH monohydrate (0.407 g, 9.71 mmol) in water (5.3 mL) was added dropwise to a solution of 3 - ((3 - ((1- (4-chlorophenyl ) Ethyl -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino) -5-methoxyphenyl) amino) cyclobutanecarboxylate 33c (600 mg, 0.971 mmol) in THF (12 mL). The mixture was stirred at room temperature for 72 h, acidified with AcOH, concentrated under reduced pressure and coevaporated twice with toluene. Purification was performed by flash chromatography on silica gel (30 µm, 24 g, CH2Cl2 / MeOH from 99/1 to 96/4). The pure fractions were combined and evaporated to dryness. A second purification was performed using Reverse Phase chromatography (Stationary phase: YMC-actus Triart-C18 10 µm 30 x 150 mm, mobile phase: 65% aqueous NH4HCO3 gradient, 0.2%, 35% CH3CN to NH4HCO3 25% aqueous, 0.2%, 75% CH3CN). The pure fractions were combined and evaporated to dryness to give 3- ((3 - ((1- (4-chlorophenyl) -2-oxo-2- (6- (trifluoromethoxy) indolin-1-yl) ethyl) amino acid ) - 5-methoxyphenyl) amino) cyclobutanecarboxylic (Compound 33, 80 mg). The stereoisomers were separated by chiral SFC (Stationary phase: Chiralcel® OD-H 5 µm 250 x 20 mm, mobile phase: 60% CO2, 40% EtOH) to give 4 fractions that were lyophilized from CH3CN / water to give Stereoisomer 33A (19 mg), Stereoisomer 33B (24 mg), Stereoisomer 33C (19 mg) and Stereoisomer 33D (26 mg). Stereoisomer 33A: 1
[00575] [00575] H NMR (500 MHz, DMSO-d6) į ppm 1.94 - 2.02 (m, 2 H) 2.43 (ddd, J = 11.8, 7.7, 4.1 Hz, 2 H) 2.91 (dt, J = 9.5, 4.7 Hz, 1 H) 3.07 -
[00576] [00576] LC / MS (LC-C method): Rt 2.86 min, MH + 590
[00577] [00577] [Į] D20: -26.2 ° (c 0.248, DMF)
[00578] [00578] Chiral SFC (SFC-I method): Rt 1.48 min, without MH +, 100% chiral purity. Stereoisomer 33B: 1
[00579] [00579] H NMR (500 MHz, DMSO-d6) į ppm 1.82 - 1.95 (m, 2 H) 2.41 - 2.47 (m, 2 H) 2.68 (tl, J = 8 , 5 Hz, 1 H) 3.08 - 3.21 (m, 2 H) 3.56 (s, 3 H) 3.62 (d ld, J = 15.4, 8.2 Hz, 1 H) 4.06 (td, J = 10.3, 7.1 Hz, 1 H) 4.46 - 4.56 (m, 1 H) 5.41 (s, 1 H) 5.43 (dl, J = 8.5 Hz, 1 H) 5.50 (s, 1 H) 5.60 (s, 2 H) 6.14 (dl, J = 8.8 Hz, 1 H) 7.01 (dl, J = 8.2 Hz, 1 H) 7.33 (d, J = 8.2 Hz, 1 H) 7.43 (d, J = 8.5 Hz, 2 H) 7.54 (d, J = 8, 5 Hz, 2 H) 8.03 (s, 1 H)
[00580] [00580] LC / MS (LC-C method): Rt 2.84 min, MH + 590
[00581] [00581] [Į] D20: -27.9 ° (c 0.248, DMF)
[00582] [00582] Chiral SFC (SFC-I method): Rt 2.20 min, without MH +, 100% chiral purity. Stereoisomer 33C: 1
[00583] [00583] H NMR (500 MHz, DMSO-d6) į ppm 1.93 - 2.03 (m, 2 H) 2.41 - 2.46 (m, 2 H) 2.82 - 2.95 (m , 1 H) 3.07 - 3.21 (m, 2 H) 3.56 (s, 3 H) 3.78 - 3.88 (m, 1 H) 4.00 - 4.11 (m, 1 H) 4.45 - 4.56 (m, 1 H) 5.37 (s, 1 H) 5.40 - 5.49 (m, 2 H) 5.58 - 5.66 (m, 2 H) 6.19 (dl, J = 8.5 Hz, 1 H) 7.00 (dl, J = 6.6 Hz, 1 H) 7.32 (dl, J = 8.2 Hz, 1 H) 7, 43 (d, J = 8.5 Hz, 2 H) 7.54 (d, J = 8.2 Hz, 2 H) 8.02 (ls, 1 H)
[00584] [00584] LC / MS (LC-C method): Rt 2.86 min, MH + 590
[00585] [00585] [Į] D20: + 26.7 ° (c 0.221, DMF)
[00586] [00586] Chiral SFC (SFC-I method): Rt 2.91 min, without MH +, 100% chiral purity. Stereoisomer 33D: 1
[00587] [00587] H NMR (500 MHz, DMSO-d6) į ppm 1.78 - 1.96 (m, 2 H) 2.40 - 2.47 (m, 2 H) 2.65 - 2.71 (m , 1H) 3.08 - 3.21 (m, 2 H) 3.56 (s, 3 H) 3.58 - 3.67 (m, 1 H) 4.06 (td, J = 10.2, 7.3 Hz, 1 H) 4.44 - 4.56 (m, 1 H) 5.38 - 5.46 (m, 2 H) 5.50 (s, 1 H) 5.60 (s, 2 H) 6.14 (dl, J = 8.8 Hz, 1 H) 7.00 (dl, J = 6.9 Hz, 1 H) 7.33 (d, J = 8.2 Hz, 1 H) 7.43 (d, J = 8.5 Hz, 2 H) 7.54 (d, J = 8.5 Hz, 2 H) 8.03 (s, 1 H)
[00588] [00588] LC / MS (LC-C method): Rt 2.84 min, MH + 590
[00589] [00589] [Į] D20: + 23.4 ° (c 0.295, DMF)
[00590] [00590] Chiral SFC (SFC-I method): Rt 5.35 min, without MH +, 100% chiral purity. Table: compounds prepared as described above Compound Structure Optical rotation Cl OMe
[00591] [00591] The antiviral activity of all compounds of the invention was tested against the DENV-2 strain 16681 which was labeled with enhanced green fluorescent protein (eGFP). The culture medium consists of a minimum essential medium supplemented with 2% heat-inactivated fetal calf serum, 0.04% gentamicin (50 mg / mL) and 2 mM L-glutamine. Vero cells, obtained from ECACC, were suspended in culture medium and 25 µL were added to 384-well plates (2500 cells / well), which already contain the antiviral compounds. Typically, these plates contain a 5-fold serial dilution of
[00592] [00592] In parallel, the toxicity of the compounds was evaluated on the same plates. As soon as the eGFP signal was read, 40 µL of ATPlite, a cell viability dye, was added to all wells of the 384-well plates. ATP is present in all metabolically active cells and the concentration decreases very quickly when the cells undergo necrosis or apoptosis. The ATPLite assay system is based on the production of light caused by the reaction of ATP with added luciferase and D-luciferin. The plates were incubated for 10 minutes at room temperature. Then, the plates were measured in a ViewLux. The semi-maximum cytotoxic concentration (CC50) was also determined, defined as the concentration required to reduce the luminescent signal by 50% compared to that of the cell control wells. Finally, the selectivity index (SI) was determined for the compounds, which was calculated as follows: SI = CC50 / EC50. Table 1: EC50, CC50 and SI for the compounds of the invention in the DENV-2 # antiviral assay of compound EC50 (µM) N CC50 (µM) N SI N 1 0.00050 3 11 3 24400 3 1A 0.018 3 9.6 3 523 3 1B 0.018 4 10 4 579 4 1C 0.000094 3 12 3 131193 3 1D 0.00023 3 12 3 57700 3 2 0.00062 3 12 3 19200 3 2A 0.00038 3 13 3 34 500 3 2B 0.019 3 11 4 565 3 3 0.00050 3 10 3 17700 3 4 0.00014 3 10 3 73900 3
[00593] [00593] Quantitative PCR assay with tetravalent reverse transcriptase (RT-qPCR)
[00594] [00594] The antiviral activity of the compounds of the invention was tested against DENV-1 strain TC974 # 666 (NCPV), DENV-2 strain 16681, DENV-3 strain H87 (NCPV) and DENV-4 strain H241
[00595] [00595] The culture medium consisted of a minimum essential medium supplemented with 2% heat-inactivated fetal calf serum, 0.04% gentamicin (50 mg / mL) and 2 mM of L-glutamine. Vero cells, obtained from ECACC, were suspended in culture medium and 75 µL / well were added to 96-well plates (10,000 cells / well), which already contain the antiviral compounds. Typically, these plates contain a 5-fold serial dilution of 9 steps of dilution of the test compound to 200 times the final concentration in 100% DMSO (500 nL; final concentration range: 25 µM - 0.000064 µM or 2 , 5 µM - 0.0000064 µM for the most active compounds). In addition, each plate contains wells that are assigned as virus controls (containing cells and viruses in the absence of compound) and cell controls (containing cells in the absence of virus and compound). As soon as the cells were added to the plates, the plates were incubated in a fully humidified incubator (37 ° C, 5% CO 2) until the next day. Dengue virus serotype 1, 2, 3 and 4 were diluted to obtain a Cp of ~ 22-24 in the assay. Therefore, 25 µL of virus suspension was added to all wells containing test compound and to the wells assigned as virus control. In parallel, 25 µL of culture medium was added to the cell controls. Then, the plates were incubated for 3 days in a fully humidified incubator (37 ° C, 5% CO 2). After 3 days, the supernatant was removed from the wells and the cells washed twice with ice-cold PBS (~ 100 µL). The cell pellets inside the 96-well plates were stored at -80 ° C for at least 1 day. The RNA was then extracted using the Cells-to-CTTM lysis kit, according to the manufacturer's guidance (Life Technologies). Cell lysates can be stored at -80 ° C or used immediately in the reverse transcription step.
[00596] [00596] In the preparation of the reverse transcription step, mixture A (table 3A) was prepared and 7.57 µL / well was distributed in a 96-well plate. After adding 5 µL of the cell lysates, a five minute denaturation step was carried out at 75 ° C (table 3B). Then, 7.43 µL of mixture B was added (table 3C) and the reverse transcription step was started (table 3D) to generate cDNA.
[00597] [00597] Finally, a mixture of RT-qPCR was prepared, mixture C (table 4A), and 22.02 µL / well was distributed in 96-well LightCycler qPCR plates to which 3 µL of cDNA was added and the qPCR was performed according to the conditions in table 4B on a LightCycler 480.
[00598] [00598] Using the LightCycler software and an internal LIMS system, dose response curves for each compound were calculated and the effective semi-maximum concentration (EC50) and the semi-maximum cytotoxic concentration (CC50) were determined (Tables 5-8). Table 3: Synthesis of cDNA using Mixture A, denaturation, Mixture B and reverse transcription. Mixture AA Plates 8 Vol. Of Samples 828 20 Reaction (µL) Volume for Mixing Item Concentration (µL) x Final Stock Unit 1 sample samples H2O Milli-Q 7.27 6019.56 R3utr425 µM 20 0.27 0.15 124 , 20 Ractin876 µM 20 0.27 0.15 124.20 Volume 7.57 mixture / well (µL) Cell lysates 5.00
B Denaturation Step: Temp Step Denaturation Time 75 ° C 5 'maintenance Maintenance 4 ° C ão
C Mixture B Samples 864 Mixing Item Concentration Volume for (µL) x Final Stock Unit 1 sample samples Buffer X 10.00 1.00 2.00 1728.0 HIFI 2 MgCl2 mM 25.00 3.50 2.80 2419 , 2 mM dNTPs 10.00 1.00 2.00 1728.0 Rnase Inhibitor U / µL 40.00 1.00 0.50 432.0 Expand RT U / µL 50.00 0.33 0.13 112, 3 Total Volume 7.43 Mixture (µL)
D Synthesis protocol cDNA Step Temp Time Transc rev 42 ° C 30 'Denaturation 99 ° C 5' Maintenance 4 ° C maintenance Table 4: Mixture and qPCR protocol.
A Mixture C Reaction Vol. Samples 833 25 (µL) Concentration Item Volume to (µL) Mixture x Final Stock Unit 1 sample samples H2O grade PCR 7.74 6447.42 Roche Mixture 2xMM X 2 1 12.50 10412.50 Roche F3utr258 µM 20 0.3 0.38 316.54 R3utr425 µM 20 0.3 0.38 316.54
P3utr343 µM 20 0.1 0.13 108.29 Factina743 µM 20 0.3 0.38 316.54 Ractin 876 µM 20 0.3 0.38 316.54 Pactina773 µM 20 0.1 0.13 108.29 Volume Mix / Tube 22.02 (µL) cDNA 3.00
B Protocol qPCR3 Step Rate Temp Time ramp pre-incub / skim 95 ° C 10 min 4.4 Denaturation 95 ° C 10 sec 4.4 annealing 58 ° C 1 min 2.2 40 cycles Elongation 72 ° C 1 sec 4, 4 Cooling 40 ° C 10 sec 1.5 Table 5: EC50, CC50 and SI for compounds against serotype 1 in RT-qPCR tests RT-qPCR serotype 1 TC974 # 666 # of EC50 compound (µM) N CC50 (µM ) N SI N 1C 0.00023 3> 2.5 3> 11000 3 1D 0.00027 3> 2.5 3> 11400 3 2A 0.00057 3> 2.5 3> 7920 3 4C 0.000066 4> 1 , 0 4> 61000 4 4D 0.00023 4> 1.0 4> 7140 4 5A 0.00058 3> 1.0 3> 1930 3 6AB 0.000022 3> 2.5 3> 133413 3 6BB 0.00013 3 > 2.5 2> 24 100 2 7B 0.00015 3> 2.5 3> 23 200 3 8B 0.000032 3> 1.0 3> 38 900 3 9B 0.00081 3> 1.0 3> 2080 3 10B 0, 00027 3> 2.5 3> 18700 3 11B 0.00020 4> 2.5 4> 11800 4 12B 0.00047 3> 2.5 3> 8660 3
RT-qPCR serotype 1 TC974 # 666 # of EC50 compound (µM) N CC50 (µM) N SI N 13B 0.00019 3> 2.5 3> 10400 3 14A 0.00026 3> 1.0 3> 6140 3 15B 0.00024 3> 1.0 2> 6280 2 17C 0.00019 3> 1.0 3> 8230 3 17D 0.00021 3> 1.0 3> 15400 3 18B 0.00055 3> 1.0 3> 2220 3 19AB 0.000021 3> 1.0 3> 56300 3 19BB 0.000091 3> 1.0 3> 13300 3 20C 0.00030 4> 1.0 4> 4540 4 20D 0.00089 4> 1.0 4 > 1820 4 21B 0.0010 3> 2.5 3> 1950 3 22AB 0.000091 3> 1.0 3> 15700 3 22BB 0.0015 3> 2.5 1> 1740 1 23B 0.000092 5> 1, 0 5> 18200 5 24B 0.0022 4> 1.0 4> 898 4 25B 0.0013 4> 1.0 4> 1210 4 27B 0.000013 4> 1.0 4> 114 234 4 28AB 0.00041 4> 1.0 4> 4260 4 28BB 0.00011 4> 1.0 4> 29800 4 29AB 0.00023 4> 1.0 4> 5680 4 29BB 0.000048 4> 1.0 4> 30400 4 30A 0.000094 5> 1.0 5> 54900 5 31A 0.00019 3> 1.0 3> 6090 3 32B 0.00011 4> 1.0 4> 10300 4 33C 0.00097 4> 1.0 4> 3500 4 33D 0 , 00048 4> 1.0 4> 7130 4 N = the number of independent experiments in which the compounds were tested.
Table 6: EC50, CC50 and SI for compounds against serotype 2 in RT-qPCR assays
RT-qPCR serotype 2 16681 # of CC50 compound EC50 (µM) N (µM) N SI N 1C 0.00018 3> 2.5 3> 15300 3 1D 0.00023 3> 2.5 3> 9200 3 2A 0, 00048 3> 2.5 3> 6500 3 4C 0.000060 4> 1.0 4> 51900 4 4D 0.00013 4> 1.0 4> 15600 4 5A 0.00042 3> 1.0 3> 3220 3 6AB 0.000017 3> 2.5 3> 187753 3 6BB 0.00012 3> 2.5 3> 23700 3 7B 0.00012 3> 2.5 3> 30600 3 8B 0.000025 3> 1.0 3> 54100 3 9B 0.00050 3> 1.0 3> 2680 3 10B 0.00011 3> 2.5 3> 27400 3 11B 0.00010 3> 2.5 3> 22400 3 12B 0.00053 3> 2.5 3 > 8920 3 13B 0.00012 3> 2.5 3> 23 900 3 14A 0.00020 3> 1.0 3> 5890 3 15B 0.00023 4> 1.0 3> 4310 3 17C 0.00011 3> 1, 0 3> 13700 3 17D 0.00010 3> 1.0 3> 24 100 3 18B 0.00034 4> 1.0 3> 4590 3 19AB 0.000014 3> 1.0 3> 92300 3 19BB 0.000078 3> 1.0 3> 14 100 3 20C 0.000071 4> 1.0 4> 19200 4 20D 0.00019 4> 1.0 4> 7660 4 21B 0.00016 3> 2.5 3> 12700 3 22AB 0.000029 4> 1.0 3> 46400 3 22BB 0.00033 3> 2.5 2> 9450 2 23B 0.000034 5> 1.0 4> 51700 4 24B 0.00026 4> 1.0 4> 11300 4 25B 0 .00027 4> 1.0 3> 6080 3 27B 0.000014 4> 1.0 4> 92800 4 28AB 0.0 0031 4> 1.0 4> 5530 4
RT-qPCR serotype 2 16681 # of CC50 compound EC50 (µM) N (µM) N SI N 28BB 0.00010 4> 1.0 4> 19900 4 29AB 0.00025 3> 1.0 3> 5180 3 29BB 0, 000025 4> 1.0 4> 60200 4 30A 0.000076 4> 1.0 4> 65700 4 31A 0.00011 3> 1.0 3> 11600 3 32B 0.000033 4> 1.0 3> 35800 3 33C 0.0010 4> 1.0 4> 4630 4 33D 0.00066 4> 1.0 4> 6620 4 N = the number of independent experiments in which the compounds were tested.
Table 7: EC50, CC50 and SI for compounds against serotype 3 in RT-qPCR assays RT-qPCR serotype 3 H87 # of CC50 compound EC50 (µM) N (µM) N SI N 1C 0.0024 4> 2, 5 4> 1300 4 1D 0.0034 4> 2.5 4> 949 4 2A 0.0061 4> 2.5 4> 390 4 4C 0.00077 4> 1.0 3> 1660 3 4D 0.0023 4> 1.0 4> 727 4 5A 0.0081 3> 1.0 3> 170 3 6AB 0.00013 3> 2.5 3> 15 300 3 6BB 0.0016 3> 2.5 3> 1320 3 7B 0.0018 3> 2.5 3> 1810 3 8B 0.00016 3> 1.0 3> 7020 3 9B 0.0080 3> 1.0 3> 203 3 10B 0.0029 3> 2.5 3> 1370 3 11B 0 , 0018 3> 2.5 3> 1650 3 12B 0.0047 4> 2.5 4> 401 4 13B 0.0019 4> 2.5 4> 1350 4 14A 0.0028 3> 1.0 3> 584 3 15B 0.0028 3> 1.0 2> 525 2
RT-qPCR serotype 3 H87 # of CC50 compound EC50 (µM) N (µM) N SI N 17C 0.0036 3> 1.0 3> 401 3 17D 0.0027 3> 1.0 3> 541 3 18B 0, 0043 3> 1.0 3> 281 3 19AB 0.00014 3> 1.0 3> 8790 3 19BB 0.00085 5> 1.0 5> 1910 5 20C 0.0034 4> 1.0 3> 249 3 20D 0.0089 4> 1.0 4> 178 4 21B 0.0065 3> 2.5 3> 357 3 22AB 0.00049 3> 1.0 3> 4430 3 22BB 0.0090 3> 2.5 3> 245 3 23B 0.00033 5> 1.0 5> 6770 5 24B 0.014 5> 1.0 5> 123 5 25B 0.0063 5> 1.0 5> 190 5 27B 0.000091 4> 1.0 4> 22 600 4 28AB 0.0053 3> 1.0 3> 277 3 28BB 0.0012 3> 1.0 3> 1680 3 29AB 0.0046 4> 1.0 4> 264 4 29BB 0.00039 3> 1.0 3 > 3050 3 30A 0.0019 3> 1.0 3> 840 3 31A 0.0015 4> 1.0 4> 791 4 32B 0.00057 3> 1.0 3> 2870 3 33C 0.011 4> 1.0 4 > 185 4 33D 0.0023 4> 1.0 4> 732 4 N = the number of independent experiments in which the compounds were tested. Table 8: EC50, CC50 and SI for compounds against serotype 4 in RT-qPCR assays RT-qPCR serotype 4 H241 # of EC50 CC50
N N SI N compound (µM) (µM) 1C 0.016 3 8.7 3 427 3 1D 0.024 3 7.3 3 280 3
RT-qPCR serotype 4 H241 # of EC50 CC50
NN SI N compound (µM) (µM) 2A 0.027 3 2.2 3 125 3 4C 0.015 3> 1.0 2> 73 2 4D 0.026 3 3.2 3 73 3 5A 0.058 3 8.0 3 206 3 6AB 0 , 0029 3> 1.0 2> 425 2 6BB 0.020 3 5.4 3 205 3 7B 0.013 3 8.8 2 582 2 8B 0.0013 3 8.7 2 8240 2 9B 0.036 3 6.7 3 372 3 10B 0.022 3> 1.0 2> 57 2 11B 0.017 3 5.7 3 273 3 12B 0.039 3 9.2 2 318 2 13B 0.015 3 3.7 2 323 2 14A 0.015 3 8.9 3 510 3 15B 0.035 3 10 , 0 3 456 3 17C 0.016 3 1.7 2 95 2 17D 0.015 3 2.9 3 168 3 18B 0.033 3 3.0 2 148 2 19AB 0.00069 3 6.4 3 8200 3 19BB 0.0068 3 5, 2 3 905 3 20C 0.020 3> 1.0 2> 60 2 20D 0.055 3> 1.0 1> 21 1 21B 0.048 3 9.0 3 115 3 22AB 0.0033 3 9.1 2 2890 2 22BB 0.071 3 8 , 2 3 126 3 23B 0.0027 3 6.0 2 2230 2 24B 0.066 3 7.8 3 115 3 25B 0.055 3 9.9 3 202 3 27B 0.00081 3 8.3 3 9330 3 28AB 0.069 3 9, 0 3 158 3 28BB 0.018 3> 1.0 2> 58 2 29AB 0.035 3 8.0 2 118 2
RT-qPCR serotype 4 H241 # of EC50 CC50
NN SI N compound (µM) (µM) 29BB 0.0028 3 8.3 3 2790 3 30A 0.018 3 7.3 3 399 3 31A 0.017 4 7.3 4 445 4 32B 0.010 3 4.3 3 295 3 33C 0 , 34 3 4.2 3 9.8 3 33D 0.022 3 5.4 3 256 3 N = the number of independent experiments in which the compounds were tested.

权利要求:
Claims (16)
[1]
1. Compound of formula (I), including any of its stereochemically isomeric forms, Cl CH3
The R3
O N A COOH (I) R1 H
N R2 characterized by the fact that R1 is trifluoromethyl, trifluoromethoxy or chlorine; R2 is hydrogen, fluorine or methoxy; R3 is hydrogen or methoxy; A represents - (CH2) n- where n is 3 or 4; -O- (CH2) n- where n is 2 or 4; -O- (CH2) n- where n is 3 and one or two CH2 are replaced by one or two CH3; -CH2-O- (CH2) n- where n is 2; or -X-Y- where X is -O-, -OCH2- or -NH-; and Y is C3-4 cycloalkyl optionally substituted by fluorine or Y is bicyclo [1.1.1] pentanil; or a pharmaceutically acceptable salt, solvate or polymorph thereof.
[2]
2. Compound according to claim 1, characterized by the fact that said compound is selected from
Cl Cl OMe OMe
O O
F O N F O N
N H N H
CUTE
F OH F OH
O O Cl Cl OMe OMe
O O
F O N F O N
N H N H
CUTE
F OH F OH
O O Cl Cl OMe OMe
THE
THE
F O N
N H N
O F O N H O
F O
F OH F
F OH
The Cl Cl OMe OMe
O O
O O
F O N F O N
N H N H
F O OH F O OH
F F Cl Cl OMe O OMe
F O
The N
N H O F O N
F N H O
F
F O
The F
OH
HO Cl Cl OMe OMe
O O
N F O N
F O N N H
H
F O O F
F F OH
OH O
Cl Cl OMe OMe
THE
THE
F O N
F O N N H
N H O O
F
F F
F OH
OH
Cl Cl MeO OMe MeO OMe
O O
F O N F O N
N H N H
CUTE
F OH F OH
O O Cl Cl OMe MeO OMe
O O
F
F N
F O N N H
N H F
O O
F O F MeO OH
OH O Cl Cl OMe OMe
O O
F F
F N F
N H N
F N H
O F O
MeO OH MeO OH
The Cl Cl OMe The OMe
F O
F N F
N H F O
F O N
F N H
OH MeO O MeO
HO Cl Cl OMe OMe
O O
THE
F O N N
N H F O N
O O H
F F O OH
F F
F OH F
Cl Cl OMe OMe
THE
THE
F N
The N H
F O N O
N H F F
The F
F O
F F
OH O
HO Cl Cl OMe OMe
O O
O O Cl N F O N
N H N H
OH OH NO
F H MeO
[3]
3. Compound according to claim 1, characterized by the fact that said compound has the specific rotation (+) measured in DMF as a solvent.
[4]
4. Compound according to claim 3, characterized by the fact that said compound is selected from: Cl Cl OMe OMe O * S O * S
F O N F O N
N H N H O * R O * S
F F
F OH F OH
O O Cl Cl OMe OMe O * R O * S
F O N F O N
N H N H
F O F O F OH F * R OH
O O Cl Cl OMe OMe O * S O * R
F O N F O N
N H N H
F O F O F * S OH F OH
O O
Cl Cl OMe OMe O * S O * S
F O N N
N H F O N O
The H * R * R
F O
F OH F
F OH
The Cl Cl OMe OMe
THE BONE
F O N
N N H
F O N H O O OH O * S * S F
F F F trans
OH Cl Cl OMe OMe
O O * S O * S
THE
F O N F O N
N H N H
F O OH F O OH
F F cis Cl Cl OMe OMe O * S O * S
F N F N
O N H O N H O cis O trans
F F
F F
O O
HO HO Cl Cl OMe OMe O * S O * S
F O N F O N
N H O N H
F O F O O
F F
OH
OH Cl Cl OMe OMe
O O * R * S
N F O N
F O N N H O
H
F F
F F
OH OH
THE
Cl Cl MeO OMe MeO OMe O * S O * S
F O N F O N
N H N H O * R O * S
F F
F OH F OH
O O Cl Cl MeO OMe MeO OMe
O O * S * S
N F O N
F O N N H
H O F O * R * R O
F
F F
OH
OH The Cl Cl MeO OMe OMe trans
O O * S F * S
N F N
F O N H O N H O * S * S F * S
F O
F MeO OH
OH trans O Cl Cl OMe OMe O O * S F * S F
F N F N
N H N H F * R F
O O MeO OH MeO OH
O O Cl Cl OMe OMe
O O F * S F * R
F N F N
F N H F N H O * R O * S O * R * S MeO OH MeO OH trans trans Cl Cl OMe OMe O * S O F O F * S
F N F
N N
F H N H
O OH F O MeO cis MeO cis O
HO
Cl Cl OMe OMe
O O * S * S
F O
F N N
F O N
N H H
F O F O OH F cis MeO F trans O
HO Cl Cl OMe OMe
O O * S trans * S trans
F O N F O N
N H F N H F
O O F O * R F O * S F * S F * R
OH OH Cl Cl OMe OMe
O O * S cis * S cis
F O N F O N
N H F N H F
O O F O * S F O * R F * S F * R
OH OH Cl Cl OMe OMe
O O * R * R
F N F N
O N H O N H
O O
F F
F F
F F cis O trans O
HO HO Cl Cl OMe OMe O * S O O * S
O Cl N F O N
N H N H
OH OH NO
F H MeO trans trans Cl OMe
O * S
THE
F O N
N H
F N OH
F H cis
[5]
5. Compound according to claim 1, characterized by the fact that R1 is trifluoromethoxy, R2 is hydrogen and R3 is hydrogen.
[6]
6. A compound according to claim 1,
characterized by the fact that A represents - (CH2) n- where n is 3 or 4.
[7]
7. A compound according to claim 1, characterized by the fact that A represents -O- (CH2) n- where n is 2 or
4.
[8]
A compound according to claim 1, characterized by the fact that A represents -O- (CH2) n- where n is 3 and one or two CH2 are replaced by one or two CH3; or A represents.
[9]
Compound according to claim 1, characterized in that A represents -X-Y- where X is an -O-, - OCH2- or -NH-; and Y is C3-4 cycloalkyl optionally substituted by fluorine.
[10]
10. Pharmaceutical composition, characterized in that it comprises a compound, as defined in any one of claims 1 to 9, together with one or more pharmaceutically acceptable excipients, diluents or carriers.
[11]
11. Pharmaceutical composition according to claim 10, characterized by the fact that it comprises a second or more active ingredients.
[12]
12. Pharmaceutical composition according to claim 11, characterized by the fact that the second or more active ingredients is an antiviral agent.
[13]
Compound of formula (I) according to any one of claims 1 to 9, characterized in that it is for use as a medicine.
[14]
A compound of formula (I) according to any one of claims 1 to 9, characterized by the fact that it is for use in the treatment of dengue infection and for the prevention or treatment of diseases associated with dengue infection.
[15]
15. Compound of formula (I) for use, according to claim 14, characterized by the fact that dengue infection is infection by virus of the strain DENV-1, DENV-2, DENV-3 or DENV-4.
[16]
16. Invention, in any form of its modalities or in any applicable class or category of claim, for example, product, process, use or sequence covered by the object initially described, disclosed or illustrated in the present patent application; A compound according to any one of claims 1 to 9, or a pharmaceutically acceptable stereoisomer thereof, for preparing a composition or medicament for preventing or treating diseases associated with dengue infection.

类似技术:

---

公开号 | 公开日 | 专利标题

---

BR112019024195A2|2020-06-23|COMPOUND AND PHARMACEUTICAL COMPOSITION OF INDOLINE DERIVATIVES REPLACED AS INHIBITORS OF DENGUE VIRAL REPLICATION

---

ES2884067T3|2021-12-10|Substituted indoline derivatives as inhibitors of dengue viral replication

---

ES2893298T3|2022-02-08|Mono- or disubstituted indole derivatives as inhibitors of dengue viral replication

---

JP6931357B2|2021-09-01|Substituted indoline derivative as a dengue virus replication inhibitor

---

ES2884151T3|2021-12-10|Substituted indoline derivatives as inhibitors of dengue virus replication

---

ES2882488T3|2021-12-02|Substituted indole derivatives as inhibitors of dengue virus replication

---

DK3201176T3|2019-03-25|MONO OR DISUBSTITUTED INDEX DERIVATIVES AS INHIBITORS OF DENGUE VIRUS REPLICATION

---

ES2887673T3|2021-12-27|Substituted indoline derivatives as inhibitors of dengue viral replication

---

BR112019024311A2|2020-07-28|substituted indoline derivatives as dengue viral replication inhibitors

---

ES2778431T3|2020-08-10|Mono- or disubstituted indole derivatives as inhibitors of dengue viral replication

---

ES2877404T3|2021-11-16|Mono or disubstituted indole derivatives as inhibitors of dengue viral replication

---

DK3201177T3|2019-03-04|MONO OR DISUBSTITUTED INDOLES AS INHIBITORS OF DENGUE VIRUS REPLICATION

---

JP6698693B2|2020-05-27|RSV antiviral pyrazolo and triazolopyrimidine compounds

---

CA3103929A1|2019-12-26|Phenyl substituted pyrazoles as modulators of roryt

---

OA19488A|2020-10-23|Substituted indoline derivatives as dengue viral replication inhibitors.

---

OA18876A|2019-09-13|Substituted indoline derivatives as dengue viral replication inhibitors

---

OA19265A|2020-06-05|Substituted indoline derivatives as dengue viral replication inhibitors

同族专利:

---

公开号 | 公开日

---

US20200255376A1|2020-08-13|

---

EP3630723A1|2020-04-08|

---

PH12019502559A1|2021-01-25|

---

WO2018215315A1|2018-11-29|

---

CR20190530A|2020-01-24|

---

AR111878A1|2019-08-28|

---

AU2018274100A1|2019-11-21|

---

TW201908291A|2019-03-01|

---

MX2019013893A|2020-01-20|

---

CA3060583A1|2018-11-29|

---

ECSP19083640A|2019-11-30|

---

MA48943A|2021-04-28|

---

EA201992782A1|2020-03-24|

---

UY37742A|2018-11-30|

---

JP2020520946A|2020-07-16|

---

CO2019012035A2|2020-01-17|

---

KR20200009027A|2020-01-29|

---

IL270726D0|2020-01-30|

---

CN110691773A|2020-01-14|

---

CL2019003294A1|2020-03-20|

---

NI201900119A|2020-03-23|

---

PE20200604A1|2020-03-10|

引用文献:

---

公开号 | 申请日 | 公开日 | 申请人 | 专利标题

---

---

US8143259B2|2008-08-19|2012-03-27|Janssen Pharmaceutica, Nv|Cold menthol receptor antagonists|

---

GB201116559D0|2011-09-26|2011-11-09|Univ Leuven Kath|Novel viral replication inhibitors|

---

EA032546B1|2014-10-01|2019-06-28|Янссен Фармасьютикалз, Инк.|Mono- or disubstituted indole derivatives as dengue viral replication inhibitors|

---

CA3013407A1|2016-03-31|2017-10-05|Janssen Pharmaceuticals, Inc.|Substituted indoline derivatives as dengue viral replication inhibitors|

---

AR108046A1|2016-04-01|2018-07-11|Janssen Pharmaceuticals Inc|INDOLINA DERIVATIVES REPLACED AS INHIBITORS OF THE VIRAL REPLICATION OF DENGUE|GB201116559D0|2011-09-26|2011-11-09|Univ Leuven Kath|Novel viral replication inhibitors|

---

US10696632B2|2015-05-08|2020-06-30|Janssen Pharmaceuticals, Inc.|Mono- or di-substituted indole derivatives as dengue viral replication inhibitors|

---

JO3633B1|2015-09-16|2020-08-27|Katholieke Univ Leuven Ku Leuven Research & Development|Mono- or di-substituted indole derivatives as dengue viral replication inhibitors|

---

KR20180052672A|2015-09-16|2018-05-18|얀센 파마슈티칼즈, 인코포레이티드|1- or 2-substituted indole derivatives as Dengue virus replication inhibitors|

---

SI3436435T1|2016-03-31|2021-08-31|Janssen Pharmaceuticals, Inc.|Substituted indole derivatives as dengue viral replication inhibitors|

---

CA3013407A1|2016-03-31|2017-10-05|Janssen Pharmaceuticals, Inc.|Substituted indoline derivatives as dengue viral replication inhibitors|

---

SG11201808237UA|2016-04-01|2018-10-30|Janssen Pharmaceuticals Inc|Substituted indole compound derivatives as dengue viral replication inhibitors|

---

AR108046A1|2016-04-01|2018-07-11|Janssen Pharmaceuticals Inc|INDOLINA DERIVATIVES REPLACED AS INHIBITORS OF THE VIRAL REPLICATION OF DENGUE|

---

JOP20180025A1|2017-03-31|2019-01-30|Janssen Pharmaceuticals Inc|Substituted indoline derivatives as dengue viral replication inhibitors|

---

JOP20180026A1|2017-03-31|2019-01-30|Univ Leuven Kath|Substituted indoline derivatives as dengue viral replication inhibitors|

---

TW201900611A|2017-05-22|2019-01-01|美商健生醫藥公司|Substituted porphyrin derivative as a dengue virus replication inhibitor |

法律状态:
2021-11-03| B350| Update of information on the portal [chapter 15.35 patent gazette]|

优先权:

---

申请号 | 申请日 | 专利标题

---

EP17172237|2017-05-22|

---

EP17172237.4|2017-05-22|

---

PCT/EP2018/063028|WO2018215315A1|2017-05-22|2018-05-18|Substituted indoline derivatives as dengue viral replication inhibitors|

---