WO2024110680A1 - Thiophene derivative compounds as antiviral agents - Google Patents
Thiophene derivative compounds as antiviral agents Download PDFInfo
- Publication number
- WO2024110680A1 WO2024110680A1 PCT/ES2023/070691 ES2023070691W WO2024110680A1 WO 2024110680 A1 WO2024110680 A1 WO 2024110680A1 ES 2023070691 W ES2023070691 W ES 2023070691W WO 2024110680 A1 WO2024110680 A1 WO 2024110680A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- thiophen
- carboxamide
- phenyl
- yloxy
- piperidin
- Prior art date
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- -1 Thiophene derivative compounds Chemical class 0.000 title claims description 142
- 239000003443 antiviral agent Substances 0.000 title description 4
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- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 claims description 20
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- DENPQNAWGQXKCU-UHFFFAOYSA-N thiophene-2-carboxamide Chemical compound NC(=O)C1=CC=CS1 DENPQNAWGQXKCU-UHFFFAOYSA-N 0.000 claims description 9
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- 208000030820 Ebola disease Diseases 0.000 claims description 5
- 125000001797 benzyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])* 0.000 claims description 5
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- 239000001257 hydrogen Substances 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000007912 intraperitoneal administration Methods 0.000 description 1
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- 229910052740 iodine Inorganic materials 0.000 description 1
- 239000011630 iodine Substances 0.000 description 1
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
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- VMGAPWLDMVPYIA-HIDZBRGKSA-N n'-amino-n-iminomethanimidamide Chemical compound N\N=C\N=N VMGAPWLDMVPYIA-HIDZBRGKSA-N 0.000 description 1
- 125000004123 n-propyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
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- 231100000065 noncytotoxic Toxicity 0.000 description 1
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- 235000019198 oils Nutrition 0.000 description 1
- 101150057545 p72 gene Proteins 0.000 description 1
- KDLHZDBZIXYQEI-UHFFFAOYSA-N palladium Substances [Pd] KDLHZDBZIXYQEI-UHFFFAOYSA-N 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 239000000312 peanut oil Substances 0.000 description 1
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- BOLDJAUMGUJJKM-LSDHHAIUSA-N renifolin D Natural products CC(=C)[C@@H]1Cc2c(O)c(O)ccc2[C@H]1CC(=O)c3ccc(O)cc3O BOLDJAUMGUJJKM-LSDHHAIUSA-N 0.000 description 1
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- QERYCTSHXKAMIS-UHFFFAOYSA-N thiophene-2-carboxylic acid Chemical compound OC(=O)C1=CC=CS1 QERYCTSHXKAMIS-UHFFFAOYSA-N 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/38—Heterocyclic compounds having sulfur as a ring hetero atom
- A61K31/381—Heterocyclic compounds having sulfur as a ring hetero atom having five-membered rings
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
- A61K31/445—Non condensed piperidines, e.g. piperocaine
- A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
- A61K31/4535—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a heterocyclic ring having sulfur as a ring hetero atom, e.g. pizotifen
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
- A61K31/5375—1,4-Oxazines, e.g. morpholine
- A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D333/00—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom
- C07D333/02—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings
- C07D333/04—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings not substituted on the ring sulphur atom
- C07D333/06—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings not substituted on the ring sulphur atom with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to the ring carbon atoms
- C07D333/08—Hydrogen atoms or radicals containing only hydrogen and carbon atoms
- C07D333/10—Thiophene
Definitions
- the present invention relates to thiophene derivatives and the use of these compounds for the prevention and/or treatment of viral diseases. More particularly, it refers to the use of said compounds in the treatment and/or prevention of the disease caused by the Ebola virus (EBOV) and/or African swine fever (ASF). Therefore, the invention falls within the field of medicinal chemistry.
- EBOV Ebola virus
- ASF African swine fever
- African swine fever In the case of veterinary infections, African swine fever is in the news, and is also one of the animal diseases with the greatest socioeconomic repercussions.
- the African swine fever virus (ASPV) produces a hemorrhagic disease with high mortality in domestic pigs and wild boars (Alejo, A.; Matamoros, T.; Guerra, M.; Andres, G. A Proteomic atlas of the African swine fever virus particle. J Virol 2018, 92, e01293-18).
- the present invention proposes new compounds derived from thiophene useful for the treatment and/or prevention of viral diseases, especially the disease caused by the Ebola virus and/or swine fever.
- the present invention presents a family of compounds that have antiviral activity in micromolar order, being useful for the treatment of viral diseases such as those caused by the Ebola virus and the African swine fever virus.
- the present invention relates to a compound of formula (I) or any of its isomers or its pharmaceutically acceptable salts: where:
- R 1 is a group selected from and -NH(CH 2 )nAr, where Bn is a benzyl group, n is an integer between 0 and 2, and Ar is an unsubstituted phenyl group (Ph) or a phenyl group substituted with a halogen or with the heterocid.
- R 2 , R 3 and R 4 are independently selected from -H, -OH, -O-C1-C3 alkyl and where at least two radicals selected from R 2 , R 3 and R 4 are H; and with the condition that if R 2 , R 3 and R 4 are -H, R 1 is and if one of the radicals selected from R 2 , R 3 and R 4 is -OH, R 1 e where the compound of formula (I) is not: for use as a medicament, or this aspect refers to the use of a compound of formula (I) as defined above for the manufacture of a medicament.
- R 1 is a group selected from NH(CH2)nAr, where n is 0 or 2.
- R 1 is a -NH(CH2)nAr group, where n is 0 or 2.
- R 1 is a group -NH(CH2)nAr where n is 0 and Ar is a phenyl unsubstituted or substituted by a chlorine.
- R 1 is a -NH(CH2)nAr group where n is 2 and Ar is an unsubstituted phenyl.
- R2 , R3 and R4 are selected independently of H
- R 4 e In another preferred embodiment, R 2 , R 3 and R 4 are -H.
- R 2 , R 3 and R 4 are independently selected from -OH and O-C1-C3 alkyl, preferably -OCH 3 , the other two substituents being H.
- R 1 is and one of R 2 , R 3 and R 4 is -O- C1-C3 alkyl, preferably -OCH3.
- the invention relates to the use of the compounds of formula (I) selected from the list comprising: N-phenyl-5-(2-(piperidin-4-yloxy)phenyl)thiophen -2-carboxamide (31) N-phenyl-5-(3-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide (32) N-phenyl-5-(4-(piperidin-4-yloxy)phenyl )thiophen-2-carboxamide (33) N-(3-chlorophenyl)-5-phenylthiophen-2-carboxamide (1 )
- a second aspect of the invention relates to the compounds of general formula (I) or any of their isomers or their pharmaceutically acceptable salts.
- R 1 , R 2 , R 3 and R 4 are as defined in the first aspect of the present invention with the condition that if R 2 , R 3 and R 4 are H, -R 1 is and if one of the radicals selected from R 2 , R 3 and R 4 is -OH, R 1 e for use in the treatment and/or prevention of viral diseases, preferably Ebola disease and African swine fever.
- the present invention also relates to the use of a compound of formula (I) as defined in the second aspect of the invention for the manufacture of a medicament for the treatment and/or prevention of viral diseases, preferably Ebola disease and of African swine fever.
- viral diseases preferably Ebola disease and of African swine fever.
- the present invention also relates to a pharmaceutical composition
- a pharmaceutical composition comprising at least one compound of formula (I) or any of its isomers or its pharmaceutically acceptable salts as defined above in the second aspect of the invention, together with a vehicle or excipient pharmaceutically acceptable for use in the treatment and/or prevention of viral diseases, preferably Ebola disease and African swine fever.
- viral diseases preferably Ebola disease and African swine fever.
- the compound must be in the therapeutically effective amount for use in the treatment and/or prevention of viral diseases. ⁇
- the therapeutically effective amount of the compound of formula (I) to be administered will depend, among other factors, on the individual (human or animal) that is going to be treated, on the severity of the disease suffered by said individual, on the form of elected administration, etc. For this reason, the doses mentioned in this invention should be considered only as guides for the person skilled in the art, and he should adjust the doses based on the variables mentioned above.
- the present invention also relates to a compound of formula (I) or any of its isomers or its pharmaceutically acceptable salts: (I) where: R 1 is a group selected d -NH(CH 2 )nAr, Bn being a benzyl group n an integer between 0 and 2 and
- R 2 , R 3 and R 4 are independently selected from -H, -OH, -OCH3 and where at least two of R 2 , R 3 and R 4 are -H; and with the condition that if R 2 , R 3 and R 4 are equal to -H, R 1 is and if one of R 2 , R 3 and R 4 is -OH, R 1 is and where the compound is not the following:
- R 1 is a group selected from NH(CH 2 )nAr, where n is 0 or 2.
- R 1 is a -NH(CH 2 )nAr group, where n is 0 or 2.
- R 1 is a group -NH(CH2)nAr where n is 0 and Ar is a phenyl unsubstituted or substituted by a chlorine.
- R 1 is a -NH(CH2)nAr group where n is 2 and Ar is an unsubstituted phenyl.
- R 2 , R 3 and R 4 are independently selected from -H, -
- R 2 is
- R 3 is
- R 4 is
- R 2 , R 3 and R 4 are H
- R 2 , R 3 and R 4 are independently selected from OH and O-C1-C3 alkyl, preferably -OCH 3 , the other two substituents being H.
- R 1 is and one of R 2 , R 3 and R 4 is an O- C1-C3 alkyl, preferably -OCH3.
- the invention relates to the compounds of formula (I) selected from the list comprising:
- a final aspect of the invention refers to a pharmaceutical composition that It comprises at least one compound of formula (I) defined in the just preceding aspect of the invention together with a pharmaceutically acceptable vehicle or excipient.
- solvate includes both pharmaceutically acceptable solvates, that is, solvates of the compound of formula (I) that can be used in the manufacture of a medicament, and pharmaceutically unacceptable solvates, which may be useful in the preparation of pharmaceutically acceptable solvates or salts.
- pharmaceutically acceptable solvate is not critical as long as it is pharmaceutically acceptable.
- the solvate is a hydrate.
- the solvates can be obtained by conventional solvation methods well known to those skilled in the art.
- the compounds of formula (I) for therapeutic use are prepared in solid form or aqueous suspension, in a pharmaceutically acceptable diluent. These preparations can be administered by any appropriate administration route, for which said preparation will be formulated in the pharmaceutical form appropriate to the chosen administration route. In a particular embodiment, the administration of the compound is carried out orally, topically, rectally or parenterally (including subcutaneous, intraperitoneal, intradermal, intramuscular, intravenous, etc.).
- the compounds described in the present invention for use, their pharmaceutically acceptable salts, solvates as well as the pharmaceutical compositions containing them can be used together with other additional drugs to provide a combination therapy.
- Said additional drugs may form part of the same pharmaceutical composition or, alternatively, may be provided in the form of a separate composition for administration simultaneous or not with that of the pharmaceutical composition comprising a compound of formula (I), or a salt or pharmaceutically acceptable solvate thereof.
- compounds of the invention for use also include compounds that differ only in the presence of one or more atoms. isotopically enriched.
- compounds that have such a structure with the exception of the substitution of a hydrogen for a deuterium or for tritium, or the substitution of a carbon for a carbon enriched in 13 C or 14 C or a nitrogen enriched in 15 N, are within of the scope of this invention.
- -Bn refers to a benzyl group, that is: -CH 2 -Ph, being
- halogen refers herein to fluorine, gold, bromine or iodine.
- the halogen proposed is chlorine.
- C1-C3 alkyl refers to linear or branched aliphatic hydrocarbon chains having 1 to 3 carbon atoms, for example, methyl, ethyl, n-propyl and i-propyl.
- treatment or prevention means to reverse, alleviate, inhibit the progress of, or prevent the disorder or condition to which it applies in such terms, one or more symptoms. of such disorder or condition.
- excipients, adjuvants and/or vehicles refers to molecular entities or substances with which the active ingredient is administered.
- excipients, adjuvants or pharmaceutical vehicles may be sterile liquids, such as waters and oils, including those of petroleum or animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like, excipients, disintegrants, wetting agents or diluents.
- Coupling constants are expressed in Hz.
- Mass spectra were acquired on a Thermo Mod. FinniganTM LXQTM spectrometer coupled to a high-performance liquid chromatograph (HPLC), using electrospray ionization (ESI).
- HPLC high-performance liquid chromatograph
- ESI electrospray ionization
- For thin layer chromatography previously coated aluminum sheets (ALUGRAM® Xtra SIL G/UV254) were used. Column chromatographies were performed in silica gel 60 (Merk) manually or automatically using the IsoleraOne equipment (Biotage®). The purity of the final compounds was verified to be >95% by HPLC analysis.
- HPLC conditions to assess purity were as follows: Surveyor HPLC equipped with a Surveyor PDA plus UV-VIS detector; ZORBAX® SB-C18 column (3.5 pm, 4.6 mm x 50 mm); gradient elution of H2O/CH3CN from 100/0 to 0/100 for 5 min; flow 800 pL/min; wavelength, UV 254 nm.
- the equipment used to determine the exact molecular mass was an Agilent 1200LC liquid chromatograph coupled to an Agilent 6500 mass spectrometer with ESI/APCI ionization source and hybrid quadrupole/time-of-flight (QTOF) analyzer located at the Spectrometry Service of Masses of the Institute of General Organic Chemistry (IQOG-CSIC).
- reaction mixture was bubbled with argon for a further 15 minutes for complete degassing and then heated under microwave irradiation (MW) for 20 minutes at 120 ° C.
- MW microwave irradiation
- the reaction crude was diluted with a 1:1 solution of AcOEt/H 2 O and extracted with AcOEt.
- the organic phase was washed with a 1:1 solution of saturated NaCl solution and H2O, and finally dried over anhydrous Na2SO4.
- the desiccant was then filtered and the volatiles evaporated to dryness under vacuum.
- the resulting residue was purified by column chromatography using solvent mixtures as eluent, indicated in each case, to obtain the desired products.
- the white solid residue obtained corresponding to the Boc-protected derivative, was dissolved in a 3:2 CH2CI2/TFA solution and stirred at room temperature for 3 hours. Once the deprotection was completed, the solvent was evaporated under vacuum and after dissolving the resulting product in CH2CI2, it was basified with NaHCO3 to obtain the product in neutral form. The volatiles were then evaporated to dryness in vacuo and the resulting residue was purified by column chromatography using the solvent mixtures indicated in each case as eluent to obtain the desired products.
- N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-(2-hydroxyphenyl)thiophen-2-carboxamide (9) (1.66 mmol, 700 mg), 4-hydroxypiperidin-1 - tere-butyl carboxylate (2.16 mmol, 434 mg), diisopropyl azodicarboxylate (2.16 mmol, 426 ⁇ L), triphenylphosphine (2.16 mmol, 434 mg), and 15 mL of THF.
- N-(4-chlorophenyl)-5-(2-hydroxyphenyl)thiophen-2-carboxamide (15) (0.76 mmol, 250 mg), tert-butyl 4-hydroxypiperidin-1-carboxylate (0.99 mmol, 199 mg), diisopropyl azodicarboxylate (0.99 mmol, 195 pL), triphenylphosphine (0.99 mmol, 259 mg) and 7 mL of THF.
- N-(4-chlorophenyl)-5-(3-hydroxyphenyl)thiophen-2-carboxamide (16) (1.71 mmol, 564 mg), tert-butyl 4-hydroxypiperidine-1-carboxylate (2.2 mmol, 447 mg), diisopropyl azodicarboxylate (2.2 mmol, 437 pL), triphenylphosphine (2.2 mmol, 582 mg) and 15 mL of THF.
- Example 2 Study of the antiviral activity of the compounds of formula (I) against the Ebola virus (IEBOV)
- HIV- May-luc recombinant viruses
- HAV human immunodeficiency virus
- HIV human immunodeficiency virus
- GP/G envelopes
- ZEBOV Ebola virus
- VSV Vesicular Stomatitis Virus
- Viral particles based on the HIV virus were generated, pseudotyped with the glycoprotein of the Ebola- Mayinga strain (GeneBank: U23187.1) (ZEBOV-GP) or the glycoprotein of the Vesicular Stomatitis Virus (VSV-G) (GenBank: X03633.1) that express the luciferase protein after infection of susceptible cells.
- G Vesicular Stomatitis Virus
- Luciferase activity was determined 48 h after infection by the luciferase assay (Luciferase Assay System, Promega, Madison, Wl) using the GloMax® Navigator Microplate Luminometer (Promega) for reading.
- VSV-Eb-luc Vesicular Stomatitis Virus
- VSV Vesicular Stomatitis Virus
- GP/G envelopes
- ZEBOV Ebola virus
- VSV Vesicular Stomatitis Virus
- BHK-21 cells were transfected with plasmids that encode the envelope protein of the Ebola virus (Mayinga strain) or the Vesicular Stomatitis Virus glycoprotein itself ( G) using Lipofectamine 3000 (Thermo Fisher Scientific). After 24 h, the cells are inoculated with recombinant pseudotypes of VSV-delta G-luc, deficient in their replicative capacity (MOI: 3-5) that contain the firefly luciferase gene in replacement of the virus's own glycoprotein. thus allowing the incorporation of heterologous glycoproteins into the new viral particles that are generated.
- MOI replicative capacity
- the viral particles are collected after 20-24 h post-inoculation, the cell debris is removed by centrifugation (1200 rpm, 10 minutes) and stored in aliquots in a -80 o C freezer.
- the calculation of the infectious titer was carried out by limiting dilution of the virus stock using Vero E6 cells as a susceptible line. Luciferase activity was determined 24 h after infection by luciferase assay (Steady- Gio® Luciferase Assay System, Promega) using the GloMax® Navigator Microplate Luminometer (Promega) for reading.
- 293T cells (2x104 cells/well) were incubated at 37 ° C for 1 h with each of the compounds and then exposed to 5000 TCID (culture infectious dose) of the above-mentioned recombinant viruses. After 48 h of incubation, cells were washed with PBS, lysed by the addition of Steady-Glo Lyssis Buffer (Promega), and light was measured on a GloMax®-Multi+ detection system (Promega, Madison, WI, USA) with the luficerase assay system (Promega, Madison, Wl).
- TCID culture infectious dose
- the recombinant VSV-G virus was used under the same conditions and in parallel to the infection with Ebola pseudotypes.
- Hela cells (2x10 4 ) treated with different concentrations were used: 10, 100 and 250 pM of each compound for 48 hours in a 96-well plate. After this period, the cells were washed with culture medium and viability was measured using the colorimetric method (Cell Titer 96 AQueous non-radioactive cell proliferation assay, Promega).
- Results at IC 50 Inhibitory concentration 50% or required to reduce viral infection by 50% in the pseudotypes indicated in each case.
- CCI 50 Cytotoxic concentration 50% or required to reduce cell growth by 50% in the type of cells indicated in each case
- C IS selectivity index (CC50/IC 50 )
- Element 3 Study of the antiviral activity of the compounds of formula (I) against the West African swine virus (ASPV)
- infections with ASFV were carried out on monolayers of Vero cells (ATCC-CCL-81; renal fibroblasts) at a multiplicity of infection (mdi) of 5 pfu/cell of the non-pathogenic ASFV Ba71 V isolate.
- Vero cells Enjuanes, L., Carrascosa, A.L., Moreno, M.A., Vinuela, E., 1976. Titration of African swine fever (ASF) virus. J Gen Virol 32, 471-477) or with fluorescent recombinant viruses for flow cytometry as described below.
- the viral inocula were always added to the minimum volume necessary to cover the cell mat, after pretreatment of the cells with the compounds for one hour at 37oC . After the pre-treatment time had elapsed, the viral inoculum was allowed to absorb for 60 minutes at 37 °C and finally, the medium was removed along with the viral inoculum and replaced with supplemented Dulbecco's modified Eagle culture medium (DMEM). with 2% fetal bovine serum (FBS) in combination with the different compounds to continue the treatment until reaching 16 hours post infection (hpi).
- DMEM Dulbecco's modified Eagle culture medium
- FBS fetal bovine serum
- DMSO dimethyl sulfoxide
- FBS fetal bovine serum
- Cell viability in the presence of the different compounds at various concentrations was determined after 24 hours using the commercial kit “Cell titer 96 Aqueous Non-Radioactive Cell Proliferation Assay” (Promega) by reading the absorbance values at 490 nm. following the manufacturer's instructions. Those concentrations that allowed cell viability equal to or greater than 80% were considered non-cytotoxic. This system is based on the colorimetric quantification of the reduction of tetrazolium to formazan by the activity of dehydrogenase enzymes of metabolically active cells.
- the cells were extracted and the subsequent DNA obtained and purified with the “Dneasy blood and tissue kit” (Qiagen). The concentrations obtained were quantified in the “UV-Vis Nanodrop ND- 1000” spectrophotometer (Fisher Termo Scientific).
- qPCR real-time quantitative PCR
- the mixture for amplification was composed of 150 ng of DNA from each of the samples, the Taqman probe (Roche), the primers OE3F and OE4R whose sequence is described by King et al., obtained from Fisher Scientific and “Premix Ex Taq (2X)” (Takara).
- the amplification and quantification process was carried out in an “ABI 7500 Fast Real-Time PCR System” thermocycler (Applied Biosystems) with the following parameters: 1 cycle of 94 o C for 10 minutes, 45 cycles of 94 o C for 15 seconds and 58 o C for 60 seconds.
- the cells were infected with the recombinant viruses made in the laboratory BPP30GFP or B54GFP, described in Barrado-Gil, L; Galindo, I.; Martinez-Alonso, D.; Viedma, S.; Alonso, C.
- the ubiquitin-proteasome system is required for African swine fever replication.
- Results at IC 50 Inhibitory concentration 50% or required to reduce viral infection by 50%.
- b CCI 50 Cytotoxic concentration 50% or required to reduce cell growth by 50%.
- C IS selectivity index (CC50/IC50).
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Abstract
The present invention relates to thiophene derivatives and to the use of these compounds of formula (I) to prevent and/or treat viral diseases. More particularly, the invention relates to the use of these compounds to treat and/or prevent the disease caused by Ebola virus (EBOV) and/or African swine fever (ASF).
Description
COMPUESTOS DERIVADOS DE TIOFENO COMO AGENTES ANTIVIRALES COMPOUNDS DERIVED FROM THIOPHENE AS ANTIVIRAL AGENTS
La presente invención se refiere a derivados de tiofenos y al uso de estos compuestos para la prevención y/o tratamiento de enfermedades víricas. De forma más particular, se refiere al uso de dichos compuestos en el tratamiento y/o prevención de la enfermedad causada por el virus del Ébola (EBOV) y/o de la peste porcina africana (PPA). Por tanto, la invención se encuadra en el campo de la química médica. The present invention relates to thiophene derivatives and the use of these compounds for the prevention and/or treatment of viral diseases. More particularly, it refers to the use of said compounds in the treatment and/or prevention of the disease caused by the Ebola virus (EBOV) and/or African swine fever (ASF). Therefore, the invention falls within the field of medicinal chemistry.
ANTECEDENTES DE LA INVENCIÓN BACKGROUND OF THE INVENTION
Actualmente, estamos asistiendo a un aumento de las enfermedades víricas emergentes y re-emergentes a nivel mundial. De hecho, las enfermedades infecciosas son una de las mayores preocupaciones de la sanidad pública tanto humana como veterinaria. Tal es el caso de la enfermedad por el virus del Ébola. Se trata de una enfermedad muy grave y contagiosa, causante de fiebre hemorrágica y con una alta tasa de mortalidad en humanos, que puede llegar a ser del 90%. A día de hoy existen pocas alternativas para el tratamiento de esta enfermedad, y debido a la gravedad de la infección y lo imprevisible de la aparición de nuevos brotes, la necesidad de desarrollar antivirales específicos es muy grande (Malvy, D.; McElroy, A. K.; de Clerck, H.; Gunther, S.; van Griensven, J. Ebola virus disease. Lancet 2019, 393, 936-948). En el caso de infecciones veterinarias, la peste porcina africana acapara la actualidad siendo además una de las enfermedades animales de mayores repercusiones socioeconómicas. El virus de la peste porcina africana (VPPA) produce una enfermedad hemorrágica de elevada mortalidad en el cerdo doméstico y en los jabalíes (Alejo, A.; Matamoros, T.; Guerra, M.; Andres, G. A Proteomic atlas of the African swine fever virus particle. J Virol 2018, 92, e01293-18). Currently, we are witnessing an increase in emerging and re-emerging viral diseases globally. In fact, infectious diseases are one of the greatest concerns of both human and veterinary public health. Such is the case of the Ebola virus disease. It is a very serious and contagious disease, causing hemorrhagic fever and with a high mortality rate in humans, which can be up to 90%. Today there are few alternatives for the treatment of this disease, and due to the severity of the infection and the unpredictability of the appearance of new outbreaks, the need to develop specific antivirals is very great (Malvy, D.; McElroy, A. K. ; de Clerck, H.; Gunther, S.; van Griensven, J. Ebola virus disease. In the case of veterinary infections, African swine fever is in the news, and is also one of the animal diseases with the greatest socioeconomic repercussions. The African swine fever virus (ASPV) produces a hemorrhagic disease with high mortality in domestic pigs and wild boars (Alejo, A.; Matamoros, T.; Guerra, M.; Andres, G. A Proteomic atlas of the African swine fever virus particle. J Virol 2018, 92, e01293-18).
A la vista de lo conocido en el estado de la técnica, sería deseable disponer de fármacos efectivos contra estas enfermedades causadas por virus. Con esa finalidad, la presente invención propone nuevos compuestos derivados de tiofeno útiles para el tratamiento y/o prevención de enfermedades víricas, especialmente de la enfermedad causada por el virus del Ébola y/o de la peste porcina. In view of what is known in the state of the art, it would be desirable to have effective drugs against these diseases caused by viruses. To this end, the present invention proposes new compounds derived from thiophene useful for the treatment and/or prevention of viral diseases, especially the disease caused by the Ebola virus and/or swine fever.
DESCRIPCIÓN DE LA INVENCIÓN
La presente invención presenta una familia de compuestos que poseen actividad antiviral en orden micromolar, resultandos útiles para el tratamiento de enfermedades víricas tales como las causadas por el virus del Ébola y el virus de la peste porcina africana. DESCRIPTION OF THE INVENTION The present invention presents a family of compounds that have antiviral activity in micromolar order, being useful for the treatment of viral diseases such as those caused by the Ebola virus and the African swine fever virus.
En un primer aspecto, la presente invención se refiere a un compuesto de fórmula (I) o cualquiera de sus isómeros o sus sales farmacéuticamente aceptables:
donde: In a first aspect, the present invention relates to a compound of formula (I) or any of its isomers or its pharmaceutically acceptable salts: where:
R1 es un grupo seleccionado de y -NH(CH2)nAr,
siendo Bn un grupo bencilo n un número entero comprendido entre 0 y 2 y Ar un grupo fenilo (Ph) sin sustituir o un grupo fenilo sustituido con un halógeno o con el heterocido
R 1 is a group selected from and -NH(CH 2 )nAr, where Bn is a benzyl group, n is an integer between 0 and 2, and Ar is an unsubstituted phenyl group (Ph) or a phenyl group substituted with a halogen or with the heterocid.
R2, R3 y R4 se seleccionan independientemente de -H, -OH, -O-alquilo C1-C3 y donde al menos dos radicales seleccionados de R2, R3 y R4 son H;
y con la condición de que si R2, R3 y R4 son -H, R1 es
y si uno de los radicales seleccionados de R2, R3 y R4 es -OH, R1 e
donde el compuesto de fórmula (I) no es:
para su uso como medicamento , o bien este aspecto se refiere al uso de un compuesto de fórmula (I) como se ha definido anteriormente para la fabricación de un medicamento. R 2 , R 3 and R 4 are independently selected from -H, -OH, -O-C1-C3 alkyl and where at least two radicals selected from R 2 , R 3 and R 4 are H; and with the condition that if R 2 , R 3 and R 4 are -H, R 1 is and if one of the radicals selected from R 2 , R 3 and R 4 is -OH, R 1 e where the compound of formula (I) is not: for use as a medicament, or this aspect refers to the use of a compound of formula (I) as defined above for the manufacture of a medicament.
/ En una realización preferida, R1 es un grupo seleccionado de NH(CH2)nAr, donde n es 0 ó 2.
/ In a preferred embodiment, R 1 is a group selected from NH(CH2)nAr, where n is 0 or 2.
En otra realización preferida, R1 es un grupo -NH(CH2)nAr, donde n es 0 ó 2. In another preferred embodiment, R 1 is a -NH(CH2)nAr group, where n is 0 or 2.
En otra realización preferida, R1 es un grupo -NH(CH2)nAr donde n es 0 y Ar es un fenilo sin sustituir o sustituido por un cloro. In another preferred embodiment, R 1 is a group -NH(CH2)nAr where n is 0 and Ar is a phenyl unsubstituted or substituted by a chlorine.
En otra realización preferida, R1 es un grupo -NH(CH2)nAr donde n es 2 y Ar es un fenilo sin sustituir. In another preferred embodiment, R 1 is a -NH(CH2)nAr group where n is 2 and Ar is an unsubstituted phenyl.
En otra realización preferida, R2, R3 y R4 se seleccionan independientemente de H,
In another preferred embodiment, R2 , R3 and R4 are selected independently of H,
En una realización preferida, R2e In a preferred embodiment, R 2 e
En una realización preferida, R3e In a preferred embodiment, R 3 e
En una realización preferida, R4e
En otra realización preferida, R2, R3 y R4 son -H. In a preferred embodiment, R 4 e In another preferred embodiment, R 2 , R 3 and R 4 are -H.
En otra realización preferida, R2, R3 y R4 se seleccionan independientemente de -OH y O-alquilo C1-C3, preferiblemente -OCH3, siendo los otros dos sustituyentes H. In another preferred embodiment, R 2 , R 3 and R 4 are independently selected from -OH and O-C1-C3 alkyl, preferably -OCH 3 , the other two substituents being H.
En otra realización preferida, R1 es y uno de R2, R3 y R4 es -O-
alquilo C1-C3, preferiblemente -OCH3. In another preferred embodiment, R 1 is and one of R 2 , R 3 and R 4 is -O- C1-C3 alkyl, preferably -OCH3.
En una realización más preferida de este primer aspecto, la invención se refiere al uso de los compuestos de fórmula (I) seleccionados de la lista que comprende: N-fenil-5-(2-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (31 ) N-fenil-5-(3-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (32) N-fenil-5-(4-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (33) N-(3-clorofenil)-5-feniltiofen-2-carboxamida (1 ) In a more preferred embodiment of this first aspect, the invention relates to the use of the compounds of formula (I) selected from the list comprising: N-phenyl-5-(2-(piperidin-4-yloxy)phenyl)thiophen -2-carboxamide (31) N-phenyl-5-(3-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide (32) N-phenyl-5-(4-(piperidin-4-yloxy)phenyl )thiophen-2-carboxamide (33) N-(3-chlorophenyl)-5-phenylthiophen-2-carboxamide (1 )
N-(2-(1 -bencilpiperidin-4-il)etil)-5-feniltiofen-2-carboxamida (2) N-(2-(1-bencilpiperidin-4-il)etil)-5-(2-hidroxifenil)tiofen-2-carboxamida (9) N-(4-clorofenil)-5-(2-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (25) N-(3-clorofenil)-5-(2-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (22) N-fenetil-5-(2-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (28) N-(4-morfolinofenil)-5-(2-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (30) N-(2-(1 -bencilpiperidin-4-il)etil)-5-(2-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (21 ) N-(3-clorofenil)-5-(4-hidroxifenil)tiofen-2-carboxamida (14) N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-phenylthiophen-2-carboxamide (2) N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-(2-hydroxyphenyl )thiophen-2-carboxamide (9) N-(4-chlorophenyl)-5-(2-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide (25) N-(3-chlorophenyl)-5-( 2-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide (22) N-phenethyl-5-(2-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide (28) N-(4- morpholinophenyl)-5-(2-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide (30) N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-(2-(piperidin- 4-yloxy)phenyl)thiophen-2-carboxamide (21 ) N-(3-chlorophenyl)-5-(4-hydroxyphenyl)thiophen-2-carboxamide (14)
(N-(2-(1 -bencilpiperidin-4-il)etil)-5-(4-hidroxifenil)tiofen-2-carboxamida (11 ) N-(3-dorofenil)-5-(3-hidroxifenil)tiofen-2-carboxamida (13) (N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-(4-hydroxyphenyl)thiophen-2-carboxamide (11) N-(3-dorophenyl)-5-(3-hydroxyphenyl)thiophen- 2-carboxamide (13)
N-(2-(1 -bencilpiperidin-4-il)etil)-5-(3-hidroxifenil)tiofen-2-carboxamida (10) N-(4-clorofenil)-5-(3-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (26) N-(3-clorofenil)-5-(3-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (23) N-fenetil-5-(3-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (29) N-(2-(1-bencilpiperidin-4-il)etil)-5-(4-metoxifenil)tiofen-2-carboxamida (35) N-(2-(1-bencilpiperidin-4-il)etil)-5-(3-metoxifenil)tiofen-2-carboxamida (34) N-(4-clorofenil)-5-(4-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (27) N-(3-clorofenil)-5-(4-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (24).
Un segundo aspecto de la invención se refiere a los compuestos de fórmula general (I) o cualquiera de sus isómeros o sus sales farmacéuticamente aceptables
donde R1, R2, R3 y R4 son como se ha definido en el primer aspecto de la presente invención con la condición de que si R2, R3 y R4 son H,-R1 es
y si uno de los radicales seleccionados de R2, R3 y R4 es -OH, R1 e
para su uso en el tratamiento y/o prevención de enfermedades víricas, preferiblemente la enfermedad del Ébola y de la peste porcina africana. N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-(3-hydroxyphenyl)thiophen-2-carboxamide (10) N-(4-chlorophenyl)-5-(3-(piperidin-4- yloxy)phenyl)thiophen-2-carboxamide (26) N-(3-chlorophenyl)-5-(3-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide (23) N-phenethyl-5-(3) -(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide (29) N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-(4-methoxyphenyl)thiophen-2-carboxamide (35) N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-(3-methoxyphenyl)thiophen-2-carboxamide (34) N-(4-chlorophenyl)-5-(4-(piperidin-4- yloxy)phenyl)thiophen-2-carboxamide (27) N-(3-chlorophenyl)-5-(4-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide (24). A second aspect of the invention relates to the compounds of general formula (I) or any of their isomers or their pharmaceutically acceptable salts. where R 1 , R 2 , R 3 and R 4 are as defined in the first aspect of the present invention with the condition that if R 2 , R 3 and R 4 are H, -R 1 is and if one of the radicals selected from R 2 , R 3 and R 4 is -OH, R 1 e for use in the treatment and/or prevention of viral diseases, preferably Ebola disease and African swine fever.
La presente invención también se refiere al uso de un compuesto de fórmula (I) como se ha definido en el segundo aspecto de la invención para la fabricación de un medicamento para el tratamiento y/o prevención de enfermedades víricas, preferiblemente la enfermedad del Ébola y de la peste porcina africana. The present invention also relates to the use of a compound of formula (I) as defined in the second aspect of the invention for the manufacture of a medicament for the treatment and/or prevention of viral diseases, preferably Ebola disease and of African swine fever.
La presente invención también se refiere a una composición farmacéutica que comprende al menos un compuesto de fórmula (I) o cualquiera de sus isómeros o sus sales farmacéuticamente aceptables como se ha definido anteriormente en el segundo aspecto de la invención, junto con un vehículo o excipiente farmacéuticamente aceptable para su uso en el tratamiento y/o prevención de enfermedades víricas, preferiblemente la enfermedad del Ébola y de la peste porcina africana. The present invention also relates to a pharmaceutical composition comprising at least one compound of formula (I) or any of its isomers or its pharmaceutically acceptable salts as defined above in the second aspect of the invention, together with a vehicle or excipient pharmaceutically acceptable for use in the treatment and/or prevention of viral diseases, preferably Ebola disease and African swine fever.
El compuesto se debe encontrar en la cantidad terapéuticamente efectiva para su uso en el tratamiento y/o prevención de enfermedades víricas.
δ The compound must be in the therapeutically effective amount for use in the treatment and/or prevention of viral diseases. δ
En general, la cantidad terapéuticamente efectiva del compuesto de fórmula (I) a administrar dependerá, entre otros factores, del individuo (humano o animal) que vaya a ser tratado, de la severidad de la enfermedad que padezca dicho individuo, de la forma de administración elegida, etc. Por este motivo, las dosis mencionadas en esta invención deben ser consideradas tan solo como guías para el experto en la materia, y éste debe ajustar las dosis en función de las variables citadas anteriormente. In general, the therapeutically effective amount of the compound of formula (I) to be administered will depend, among other factors, on the individual (human or animal) that is going to be treated, on the severity of the disease suffered by said individual, on the form of elected administration, etc. For this reason, the doses mentioned in this invention should be considered only as guides for the person skilled in the art, and he should adjust the doses based on the variables mentioned above.
La presente invención también se refiere a un compuesto de fórmula (I) o cualquiera de sus isómeros o sus sales farmacéuticamente aceptables:
(I) donde: R1 es un grupo seleccionado d -NH(CH2)nAr,
siendo Bn un grupo bencilo n un número entero comprendido entre 0 y 2 y The present invention also relates to a compound of formula (I) or any of its isomers or its pharmaceutically acceptable salts: (I) where: R 1 is a group selected d -NH(CH 2 )nAr, Bn being a benzyl group n an integer between 0 and 2 and
Ar un grupo fenilo (Ph) sin sustituir o un grupo fenilo sustituido con un halógeno o con el heterocido
Ar an unsubstituted phenyl group (Ph) or a phenyl group substituted with a halogen or with the heterocid
R2, R3 y R4 se seleccionan independientemente de -H, -OH, -OCH3 y
donde al menos dos de R2, R3 y R4 son -H; y con la condición de que si R2, R3 y R4son iguales a -H, R1 es
y si uno de R2, R3 y R4 es -OH, R1 es
y donde el compuesto no es el siguiente:
En una realización preferida, R1 es un grupo seleccionado de NH(CH2)nAr, donde n es 0 ó 2.
R 2 , R 3 and R 4 are independently selected from -H, -OH, -OCH3 and where at least two of R 2 , R 3 and R 4 are -H; and with the condition that if R 2 , R 3 and R 4 are equal to -H, R 1 is and if one of R 2 , R 3 and R 4 is -OH, R 1 is and where the compound is not the following: In a preferred embodiment, R 1 is a group selected from NH(CH 2 )nAr, where n is 0 or 2.
En otra realización preferida, R1 es un grupo -NH(CH2)nAr, donde n es 0 ó 2. In another preferred embodiment, R 1 is a -NH(CH 2 )nAr group, where n is 0 or 2.
En una realización preferida, R1 es un grupo -NH(CH2)nAr donde n es 0 y Ar es un fenilo sin sustituir o sustituido por un cloro. In a preferred embodiment, R 1 is a group -NH(CH2)nAr where n is 0 and Ar is a phenyl unsubstituted or substituted by a chlorine.
En otra realización preferida, R1 es un grupo -NH(CH2)nAr donde n es 2 y Ar es un fenilo sin sustituir. In another preferred embodiment, R 1 is a -NH(CH2)nAr group where n is 2 and Ar is an unsubstituted phenyl.
En otra realización preferida, R2, R3 y R4 se seleccionan independientemente de -H, - In another preferred embodiment, R 2 , R 3 and R 4 are independently selected from -H, -
OH, - OCH3 y
OH, - OCH 3 and
En una realización preferida, R2es In a preferred embodiment, R 2 is
En una realización preferida, R3es In a preferred embodiment, R 3 is
En una realización preferida, R4es
In a preferred embodiment, R 4 is
En otra realización preferida, R2, R3 y R4 son H
En otra realización preferida, R2, R3 y R4 se seleccionan independientemente de OH y O-alquilo C1-C3, preferiblemente -OCH3 , siendo los otros dos sustituyentes H. En otra realización preferida, R1 es y uno de R2, R3 y R4 es un O-
alquilo C1-C3, preferiblemente -OCH3 . In another preferred embodiment, R 2 , R 3 and R 4 are H In another preferred embodiment, R 2 , R 3 and R 4 are independently selected from OH and O-C1-C3 alkyl, preferably -OCH 3 , the other two substituents being H. In another preferred embodiment, R 1 is and one of R 2 , R 3 and R 4 is an O- C1-C3 alkyl, preferably -OCH3.
En una realización más preferida, la invención se refiere a los compuestos de fórmula (I) seleccionados de la lista que comprende: In a more preferred embodiment, the invention relates to the compounds of formula (I) selected from the list comprising:
N-fenil-5-(2-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (31 ) N-phenyl-5-(2-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide (31)
N-fenil-5-(3-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (32) N-phenyl-5-(3-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide (32)
N-fenil-5-(4-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (33) N-phenyl-5-(4-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide (33)
N-(2-(1 -bencilpiperidin-4-il)etil)-5-feniltiofen-2-carboxamida (2) N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-phenylthiophen-2-carboxamide (2)
N-(2-(1-bencilpiperidin-4-il)etil)-5-(2-hidroxifenil)tiofen-2-carboxamida (9) N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-(2-hydroxyphenyl)thiophen-2-carboxamide (9)
N-(4-clorofenil)-5-(2-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (25) N-(4-chlorophenyl)-5-(2-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide (25)
N-(3-clorofenil)-5-(2-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (22) N-(3-chlorophenyl)-5-(2-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide (22)
N-fenetil-5-(2-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (28) N-phenethyl-5-(2-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide (28)
N-(4-morfolinofenil)-5-(2-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (30) N-(4-morpholinophenyl)-5-(2-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide (30)
N-(2-(1 -bencilpiperidin-4-il)etil)-5-(2-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (21 )N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-(2-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide (21)
N-(3-clorofenil)-5-(4-hidroxifenil)tiofen-2-carboxamida (14) (N-(2-(1 -bencilpiperidin-4-il)etil)-5-(4-hidroxifenil)tiofen-2-carboxamida (11 ) N-(3-chlorophenyl)-5-(4-hydroxyphenyl)thiophen-2-carboxamide (14) (N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-(4-hydroxyphenyl)thiophen- 2-carboxamide (11)
N-(3-clorofenil)-5-(3-hidroxifenil)tiofen-2-carboxamida (13) N-(3-chlorophenyl)-5-(3-hydroxyphenyl)thiophen-2-carboxamide (13)
N-(2-(1 -bencilpiperidin-4-il)etil)-5-(3-hidroxifenil)tiofen-2-carboxamida (10) N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-(3-hydroxyphenyl)thiophen-2-carboxamide (10)
N-(4-clorofenil)-5-(3-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (26) N-(4-chlorophenyl)-5-(3-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide (26)
N-(3-clorofenil)-5-(3-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (23) N-(3-chlorophenyl)-5-(3-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide (23)
N-fenetil-5-(3-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (29) N-phenethyl-5-(3-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide (29)
N-(2-(1-bencilpiperidin-4-il)etil)-5-(4-metoxifenil)tiofen-2-carboxamida (35) N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-(4-methoxyphenyl)thiophen-2-carboxamide (35)
N-(2-(1-bencilpiperidin-4-il)etil)-5-(3-metoxifenil)tiofen-2-carboxamida (34) N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-(3-methoxyphenyl)thiophen-2-carboxamide (34)
N-(4-clorofenil)-5-(4-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (27) N-(4-chlorophenyl)-5-(4-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide (27)
N-(3-clorofenil)-5-(4-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (24) N-(3-chlorophenyl)-5-(4-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide (24)
Un último aspecto de la invención se refiere a una composición farmacéutica que
comprende al menos un compuesto de fórmula (I) definido en el aspecto justo anterior de la invención junto con un vehículo o excipiente farmacéuticamente aceptable. A final aspect of the invention refers to a pharmaceutical composition that It comprises at least one compound of formula (I) defined in the just preceding aspect of the invention together with a pharmaceutically acceptable vehicle or excipient.
Los compuestos de la invención y compuestos para su uso representados por la fórmula (I) pueden estar en forma cristalina como compuestos libres o como solvatos y se pretende que ambas formas estén dentro del alcance de la presente invención. En este sentido, el término “solvato”, tal como aquí se utiliza, incluye tanto solvatos farmacéuticamente aceptables, es decir, solvatos del compuesto de fórmula (I) que pueden ser utilizados en la elaboración de un medicamento, como solvatos farmacéuticamente no aceptables, los cuales pueden ser útiles en la preparación de solvatos o sales farmacéuticamente aceptables. La naturaleza del solvato farmacéuticamente aceptable no es crítica siempre y cuando sea farmacéuticamente aceptable. En una realización particular, el solvato es un hidrato. Los solvatos pueden obtenerse por métodos convencionales de solvatación bien conocidos por los técnicos en la materia. The compounds of the invention and compounds for use represented by formula (I) may be in crystalline form as free compounds or as solvates and both forms are intended to be within the scope of the present invention. In this sense, the term "solvate", as used herein, includes both pharmaceutically acceptable solvates, that is, solvates of the compound of formula (I) that can be used in the manufacture of a medicament, and pharmaceutically unacceptable solvates, which may be useful in the preparation of pharmaceutically acceptable solvates or salts. The nature of the pharmaceutically acceptable solvate is not critical as long as it is pharmaceutically acceptable. In a particular embodiment, the solvate is a hydrate. The solvates can be obtained by conventional solvation methods well known to those skilled in the art.
Los compuestos de fórmula (I) para su uso terapéutico se preparan en forma sólida o suspensión acuosa, en un diluyente farmacéuticamente aceptable. Estos preparados pueden ser administrados por cualquier vía de administración apropiada, para lo cual dicho preparado se formulará en la forma farmacéutica adecuada a la vía de administración elegida. En una realización particular, la administración del compuesto se efectúa por vía oral, tópica, rectal o parenteral (incluyendo subcutánea, intraperitoneal, intradérmica, intramuscular, intravenosa, etc.). The compounds of formula (I) for therapeutic use are prepared in solid form or aqueous suspension, in a pharmaceutically acceptable diluent. These preparations can be administered by any appropriate administration route, for which said preparation will be formulated in the pharmaceutical form appropriate to the chosen administration route. In a particular embodiment, the administration of the compound is carried out orally, topically, rectally or parenterally (including subcutaneous, intraperitoneal, intradermal, intramuscular, intravenous, etc.).
Los compuestos descritos en la presente invención para su uso, sus sales farmacéuticamente aceptables, solvatos así como las composiciones farmacéuticas que los contienen pueden ser utilizados junto con otros fármacos adicionales para proporcionar una terapia de combinación. Dichos fármacos adicionales pueden formar parte de la misma composición farmacéutica o, alternativamente, pueden ser proporcionados en forma de una composición separada para su administración simultánea o no a la de la composición farmacéutica que comprende un compuesto de fórmula (I), o una sal o solvato farmacéuticamente aceptables del mismo. The compounds described in the present invention for use, their pharmaceutically acceptable salts, solvates as well as the pharmaceutical compositions containing them can be used together with other additional drugs to provide a combination therapy. Said additional drugs may form part of the same pharmaceutical composition or, alternatively, may be provided in the form of a separate composition for administration simultaneous or not with that of the pharmaceutical composition comprising a compound of formula (I), or a salt or pharmaceutically acceptable solvate thereof.
A menos que se indique lo contrario, los compuestos de la invención para su uso también incluyen compuestos que difieren sólo en la presencia de uno o más átomos
isotópicamente enriquecidos. Por ejemplo, compuestos que tienen dicha estructura, a excepción de la sustitución de un hidrógeno por un deuterio o por tritio, o la sustitución de un carbono por un carbono enriquecido en 13C o 14C o un nitrógeno enriquecido en 15N, están dentro del alcance de esta invención. Unless otherwise indicated, compounds of the invention for use also include compounds that differ only in the presence of one or more atoms. isotopically enriched. For example, compounds that have such a structure, with the exception of the substitution of a hydrogen for a deuterium or for tritium, or the substitution of a carbon for a carbon enriched in 13 C or 14 C or a nitrogen enriched in 15 N, are within of the scope of this invention.
En la presente invención, “-Bn” se refiere a un grupo bencilo, esto es: -CH2-Ph, siendo In the present invention, “-Bn” refers to a benzyl group, that is: -CH 2 -Ph, being
Ph un grupo fenilo
Ph a phenyl group
El término “halógeno” se refiere en la presente invención a flúor, doro, bromo o yodo. En la presente invención, el halógeno proferido es el cloro. The term "halogen" refers herein to fluorine, gold, bromine or iodine. In the present invention, the halogen proposed is chlorine.
El término “alquilo C1-C3” se refiere a cadenas hidrocarbonadas alifáticas, lineales o ramificadas, que tienen de 1 a 3 átomos de carbono, por ejemplo, metilo, etilo, n-propilo e i-propilo. The term "C1-C3 alkyl" refers to linear or branched aliphatic hydrocarbon chains having 1 to 3 carbon atoms, for example, methyl, ethyl, n-propyl and i-propyl.
La expresión “tratamiento o prevención” tal y como se usa aquí, a menos que se indique lo contrario, significa revertir, aliviar, inhibir el progreso de, o prevenir el trastorno o afección al que se aplica en tales términos, uno o más síntomas de tal trastorno o afección. The term “treatment or prevention” as used herein, unless otherwise indicated, means to reverse, alleviate, inhibit the progress of, or prevent the disorder or condition to which it applies in such terms, one or more symptoms. of such disorder or condition.
La expresión “excipientes, adyuvantes y/o vehículos” se refiere a entidades moleculares o sustancias con las que se administra el ingrediente activo. Tales excipientes, adyuvantes o vehículos farmacéuticos pueden ser líquidos estériles, tales como aguas y aceites, incluyendo aquellas de petróleo o de origen animal, vegetal o sintético, tales como aceite de cacahuete, aceite de soja, aceite mineral, aceite de sésamo y similares, excipientes, disgregantes, agentes humectantes o diluyentes. The term “excipients, adjuvants and/or vehicles” refers to molecular entities or substances with which the active ingredient is administered. Such excipients, adjuvants or pharmaceutical vehicles may be sterile liquids, such as waters and oils, including those of petroleum or animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like, excipients, disintegrants, wetting agents or diluents.
A lo largo de la descripción y las reivindicaciones la palabra "comprende" y sus variantes no pretenden excluir otras características técnicas, aditivos, componentes o pasos. Para los expertos en la materia, otros objetos, ventajas y características de la invención se desprenderán en parte de la descripción y en parte de la práctica de la invención. Los siguientes ejemplos se proporcionan a modo de ilustración, y no se pretende que sean limitativos de la presente invención.
EJEMPLOS Throughout the description and claims the word "comprises" and its variants are not intended to exclude other technical characteristics, additives, components or steps. For those skilled in the art, other objects, advantages and features of the invention will emerge partly from the description and partly from the practice of the invention. The following examples are provided by way of illustration, and are not intended to be limiting of the present invention. EXAMPLES
A continuación, se ilustrará la invención mediante unos ensayos realizados por los inventores, que ponen de manifiesto la efectividad como antivirales de los compuestos de la invención. Next, the invention will be illustrated through tests carried out by the inventors, which demonstrate the effectiveness of the compounds of the invention as antivirals.
Ejemplo 1. Síntesis de compuestos de fórmula (I): Example 1. Synthesis of compounds of formula (I):
Se han sintetizado los compuestos finales indicados en la Tabla 1 , cuya actividad antiviral ha sido evaluada:
The final compounds indicated in Table 1 have been synthesized, whose antiviral activity has been evaluated:
Procedimientos experimentales. Se utilizaron disolventes de calidad analítica para todas las reacciones. Se utilizó argón para llevar a cabo las reacciones en una atmósfera inerte. Los reactivos utilizados para las reacciones fueron adquiridos en Sigma Aldrich o Fluorochem. Las reacciones por microondas se llevaron a cabo con un dispositivo InitiatorTM (Biotage®). Los puntos de fusión se registraron con un aparato Büchi Melting point M-560. Los espectros de RMN de 1H y 13C se registraron en un espectrómetro Bruker AVI II 300 MHz BACS60 o Bruker AV 500 MHz situados en el Centro de apoyo a la investigación de la Universidad Complutense de Madrid. Las abreviaturas utilizadas son s = singlete, d = doblete, t = triplete, q = cuadruplete, m = multiplete. Las constantes de acoplamiento (J) se expresan en Hz. Los espectros de masas se adquirieron en un espectrómetro Thermo Mod. FinniganTM LXQTM acoplado a un cromatógrafo líquido de alto rendimiento (HPLC), utilizando ionización por electrospray (ESI). Para la cromatografía en capa fina se utilizaron láminas de aluminio previamente recubiertas (ALUGRAM® Xtra SIL G/UV254). Las cromatografías en columna se realizaron en gel de sílice 60 (Merk) de forma manual o automática utilizando el equipo IsoleraOne (Biotage®). Se verificó que la pureza de los compuestos finales era >95% mediante análisis por HPLC. Las condiciones de HPLC para evaluar la pureza fueron las siguientes: HPLC Surveyor equipado con un PDA Surveyor más detector UV-VIS; columna ZORBAX® SB-C18 (3,5 pm, 4,6 mm x 50 mm); elución en gradiente de H2O/CH3CN de 100/0 a 0/100 durante 5 min; flujo 800 pL/min; longitud de onda, UV 254 nm. El equipo utilizado para determinar la masa molecular exacta fue un cromatógrafo de líquidos Agilent 1200LC acoplado a un espectrómetro de masas Agilent 6500 con fuente de ionización ESI/APCI y analizador de híbrido cuadrupolo/tiempo de vuelo (QTOF) situado en el Servicio de Espectrometría de Masas del Instituto de Química Orgánica General (IQOG-CSIC). Experimental procedures. Analytical grade solvents were used for all reactions. Argon was used to carry out the reactions in an inert atmosphere. The reagents used for the reactions were purchased from Sigma Aldrich or Fluorochem. Microwave reactions were carried out with an InitiatorTM device (Biotage®). The melting points were recorded with a Büchi Melting point M-560 device. The 1 H and 13 C NMR spectra were recorded on a Bruker AVI II 300 MHz BACS60 or Bruker AV 500 MHz spectrometer located at the Research Support Center of the Complutense University of Madrid. The abbreviations used are s = singlet, d = doublet, t = triplet, q = quadruplet, m = multiplet. Coupling constants (J) are expressed in Hz. Mass spectra were acquired on a Thermo Mod. FinniganTM LXQTM spectrometer coupled to a high-performance liquid chromatograph (HPLC), using electrospray ionization (ESI). For thin layer chromatography, previously coated aluminum sheets (ALUGRAM® Xtra SIL G/UV254) were used. Column chromatographies were performed in silica gel 60 (Merk) manually or automatically using the IsoleraOne equipment (Biotage®). The purity of the final compounds was verified to be >95% by HPLC analysis. HPLC conditions to assess purity were as follows: Surveyor HPLC equipped with a Surveyor PDA plus UV-VIS detector; ZORBAX® SB-C18 column (3.5 pm, 4.6 mm x 50 mm); gradient elution of H2O/CH3CN from 100/0 to 0/100 for 5 min; flow 800 pL/min; wavelength, UV 254 nm. The equipment used to determine the exact molecular mass was an Agilent 1200LC liquid chromatograph coupled to an Agilent 6500 mass spectrometer with ESI/APCI ionization source and hybrid quadrupole/time-of-flight (QTOF) analyzer located at the Spectrometry Service of Masses of the Institute of General Organic Chemistry (IQOG-CSIC).
Síntesis de 5-fenlltlofen-2-carboxamldas (1-3) v 5-bromotlofen-2-carboxamldas (4- 8)
Synthesis of 5-fenlltlofen-2-carboxymildes (1-3) and 5-bromotlofen-2-carboxymildes (4-8)
(4-8) (4-8)
Esquema 1. Procedimiento general de síntesis. Reactivos y condiciones: i) EDC, Et3N, HOBt, CH2CI2, 0 °C a 25 °C, 24h. Scheme 1. General synthesis procedure. Reagents and conditions: i) EDC, Et3N, HOBt, CH2CI2, 0 °C to 25 °C, 24h.
Procedimiento general: se añadió lentamente una solución de la amina correspondiente (1 ,02 equiv.) en CH2CI2 a una solución del ácido 2-tiofeno-carboxílico adecuado (1 equiv.), 1-etil-3-(3-dimetilaminopropil)carbodiimida (EDC) (1 ó 2 equiv.) yEt3N (2 ó 3 equiv.) en CH2CI2 a 0ºC . Para algunos derivados se añadió también 1 - hidroxibenzotriazol (HOBt) (1 equiv.). Tras agitar la mezcla de reacción durante toda la noche a temperatura ambiente, el crudo se lavó con H2O, disolución saturada de NH4CI, y disolución saturada de cloruro sódico para finalmente secarla sobre Na2SO4 anhidro. A continuación, el desecante se filtró y los volátiles se evaporaron hasta sequedad a vacío. El residuo resultante se purificó mediante cromatografía en columna flash utilizando como eluyente las mezclas de disolventes indicadas en cada caso para obtener los productos deseados. General procedure: A solution of the corresponding amine (1.02 equiv.) in CH2Cl2 was slowly added to a solution of the appropriate 2-thiophene-carboxylic acid (1 equiv.), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) (1 or 2 equiv.) yEt3N (2 or 3 equiv.) in CH2CI2 at 0 º C. For some derivatives, 1-hydroxybenzotriazole (HOBt) (1 equiv.) was also added. After stirring the reaction mixture overnight at room temperature, the crude oil was washed with H2O, saturated NH4CI solution, and saturated sodium chloride solution and finally dried over anhydrous Na2SO4. The desiccant was then filtered and the volatiles evaporated to dryness under vacuum. The resulting residue was purified by flash column chromatography using the solvent mixtures indicated in each case as eluent to obtain the desired products.
N-(3-Clorofenll)-5-fenlltlofen-2-carboxamlda (1 ) N-(3-Chlorofenll)-5-fenlltlofen-2-carboxamld (1)
Reactivos: Ácido 5-feniltiofen-2-carboxílico (1 ,5 mmol, 306 mg), 3-cloroanilina (1 ,53 mmol, 195 mg), EDC (3 mmol, 573 mg), Et3N (4,5 mmol, 626 pL) y CH2CI2 (6,0 mL). Purificación: CH2CI2/MeOH (9:1). Rendimiento: 69 mg, 15%. Sólido beige. Pf: 155 - 157 °C. 1H RMN (300 MHz, DMSO-d6) 10 δ,39 (s, 1 H), 8,04 (d, J = 4,0 Hz, 1 H), 7,93 (t, J = 2,0 Hz, 1 H), 7,73-7,81 (m, 2H), 7,69 (ddd, J = 8,3, 2,0, 1 ,0, 1 H), 7,65 (d, J = 4,0 Hz, 1 H), 7,51 - 7,37 (m, 4H), 7,18 (ddd, J = 8,0, 2,1 , 0,9 Hz, 1 H). 13C RMN (75 MHz, DMSO-d6) 6 159,9, 148,8, 140,1 , 138,2, 132,9, 132,9, 130,6, 130,4, 129,3 (2C), 128,8, 125,8 (2C), 124,5, 123,4, 119,6, 118,5. HRMS (ESI) m/z: [M + H]+ cale, para C17H13CINOS 314,0401 ; encontrado 314,0400. Pureza (HPLC): 97,8 %. Reagents: 5-Phenylthiophen-2-carboxylic acid (1.5 mmol, 306 mg), 3-chloroaniline (1.53 mmol, 195 mg), EDC (3 mmol, 573 mg), Et3N (4.5 mmol, 626 pL) and CH2CI2 (6.0 mL). Purification: CH2CI2/MeOH (9:1). Yield: 69 mg, 15%. Solid beige. mp: 155 - 157 °C. 1 H NMR (300 MHz, DMSO-d6) 10 δ.39 (s, 1 H), 8.04 (d, J = 4.0 Hz, 1 H), 7.93 (t, J = 2.0 Hz, 1 H), 7.73-7.81 (m, 2H), 7.69 (ddd, J = 8.3, 2.0, 1 .0, 1 H), 7.65 (d, J = 4.0 Hz, 1 H), 7.51 - 7.37 (m, 4H), 7.18 (ddd, J = 8.0, 2.1, 0.9 Hz, 1 H). 13 C NMR (75 MHz, DMSO-d6) 6 159.9, 148.8, 140.1, 138.2, 132.9, 132.9, 130.6, 130.4, 129.3 (2C) , 128.8, 125.8 (2C), 124.5, 123.4, 119.6, 118.5. HRMS (ESI) m/z: [M + H] + cale, for C17H13CINOS 314.0401 ; found 314.0400. Purity (HPLC): 97.8%.
N-(2-(1 -Bencllplperidln-4-ll)etll)-5-fenlltlofen-2-carboxamlda (2)
Reactivos: Ácido 5-feniltiofen-2-carboxílico (1 ,5 mmol, 306 mg), 2-(1 -bencilpiperidin-4- il)etan-1 -amina (1 ,53 mmol, 334 mg), EDC (3 mmol, 573 mg), Et3N (4,5 mmol, 626 pL) y CH2CI2 (6,0 mL). Purificación: CH2CI2/MeOH (9:1). Rendimiento: 108 mg, 18%. Sólido blanco. Pf: 150 - 152 °C. 1H RMN (300 MHz, DMSO-d6) δ 8,48 (t, J = 5,6 Hz, 1 H), 7,74 (d, J = 3,9 Hz, 1 H), 7,72 - 7,68 (m, 2H), 7,52 (d, J = 3,9 Hz, 1 H), 7,49 - 7,41 (m, 2H), 7,40 -7,21 (m, 6H), 3,47 (s, 2H), 3,27 (q, J = 7,05 Hz, 2H), 2,81 (d, J = 11 ,0 Hz, 2H), 1 ,95 (m, 2H), 1 ,69 (d, J = 11 ,4 Hz, 2H), 1 ,46 (q, J = 7,05 Hz, 2H), 1 ,39 - 1 ,26 (m, 1 H), 1 ,18 (tt, J = 5,4, 11 ,2 Hz, 2H). 13C RMN (75 MHz, DMSO-d6) δ 160,7, 147,0, 139,2, 133,1 , 129,2 (2C), 128,9, 128,8, 128,4 (2C), 128,1 (3C), 126,9, 125,6 (2C), 124,2, 62,3, 53,1 (2C), 36,8, 35,9, 32,8, 31 ,7 (2C). HRMS (ESI) m/z: [M + H]+ cale, para C25H29N2OS 405,1995; encontrado 405,1996. Pureza (HPLC) > 99 %. N-(2-(1 -Bencllplperidln-4-ll)etll)-5-fenlltlofen-2-carboxamld (2) Reagents: 5-Phenylthiophen-2-carboxylic acid (1.5 mmol, 306 mg), 2-(1-benzylpiperidin-4-yl)ethane-1-amine (1.53 mmol, 334 mg), EDC (3 mmol , 573 mg), Et3N (4.5 mmol, 626 pL) and CH 2 CI 2 (6.0 mL). Purification: CH2CI2/MeOH (9:1). Yield: 108 mg, 18%. White solid. mp: 150 - 152 °C. 1 H NMR (300 MHz, DMSO-d6) δ 8.48 (t, J = 5.6 Hz, 1 H), 7.74 (d, J = 3.9 Hz, 1 H), 7.72 - 7.68 (m, 2H), 7.52 (d, J = 3.9 Hz, 1 H), 7.49 - 7.41 (m, 2H), 7.40 -7.21 (m, 6H ), 3.47 (s, 2H), 3.27 (q, J = 7.05 Hz, 2H), 2.81 (d, J = 11.0 Hz, 2H), 1.95 (m, 2H ), 1 .69 (d, J = 11 .4 Hz, 2H), 1 .46 (q, J = 7.05 Hz, 2H), 1 .39 - 1 .26 (m, 1 H), 1 , 18 (tt, J = 5.4, 11 .2 Hz, 2H). 13 C NMR (75 MHz, DMSO-d6) δ 160.7, 147.0, 139.2, 133.1, 129.2 (2C), 128.9, 128.8, 128.4 (2C), 128.1 (3C), 126.9, 125.6 (2C), 124.2, 62.3, 53.1 (2C), 36.8, 35.9, 32.8, 31.7 (2C) ). HRMS (ESI) m/z: [M + H] + cale, for C25H29N2OS 405,1995; found 405,1996. Purity (HPLC) > 99%.
N-(2-(1 -Bencllplperldln-4-ll)etll)-5-bromotlofen-2-carboxamlda (3) N-(2-(1 -Bencllplperldln-4-ll)etll)-5-bromotlofen-2-carboxymilda (3)
Ácido 5-bromo-2-tiofencarboxílico (2 mmol, 414 mg), 2-(1 -benc¡lpiperid¡n-4-il)etan-1 - amina (2,04 mmol, 445 mg), EDC (2 mmol, 310 mg),Et3N (4 mmol, 557 pL), HOBt (2 mmol, 270 mg) y CH2CI2 (6,0 mL). El producto se usó en el paso siguiente sin purificación posterior. Rendimiento: 643 mg, 79%. Sólido blanco. 1H NMR (300 MHz, DMSO-d6) 6 8,51 (t, J = 5,6 Hz, 1 H), 7,56 (d, J = 4,0 Hz, 1 H), 7,34 - 7,21 (m, 7H), 3,42 (s, 2H), 3,24 (q, J = 6,7 Hz, 2H), 2,83 - 2,71 (m, 2H), 1 ,94 - 1 ,82 (m, 2H), 1 ,69 - 1 ,60 (m, 2H), 1 ,47 - 1 ,39 (m, 2H), 1 ,33 - 1 ,22 (m, 1 H), 1 ,22 - 1 ,08 (m, 2H). 13C NMR (75 MHz, DMSO-d6) 6 159,8, 142,1 , 138,7, 131 ,4, 128,7 (2C), 128,4, 128,1 (2C), 126,7, 116,4, 62,5, 53,2 (2C), 36,8, 35,8, 32,9, 31 ,8 (2C). 5-Bromo-2-thiophencarboxylic acid (2 mmol, 414 mg), 2-(1-benzylpiperidín-4-yl)ethane-1 -amine (2.04 mmol, 445 mg), EDC (2 mmol , 310 mg), Et3N (4 mmol, 557 pL), HOBt (2 mmol, 270 mg) and CH2CI2 (6.0 mL). The product was used in the next step without further purification. Yield: 643 mg, 79%. White solid. 1 H NMR (300 MHz, DMSO-d6) 6 8.51 (t, J = 5.6 Hz, 1 H), 7.56 (d, J = 4.0 Hz, 1 H), 7.34 - 7.21 (m, 7H), 3.42 (s, 2H), 3.24 (q, J = 6.7 Hz, 2H), 2.83 - 2.71 (m, 2H), 1 .94 - 1 .82 (m, 2H), 1 .69 - 1 .60 (m, 2H), 1 .47 - 1 .39 (m, 2H), 1 .33 - 1 .22 (m, 1 H), 1 .22 - 1 .08 (m, 2H). 13 C NMR (75 MHz, DMSO-d6) 6 159.8, 142.1, 138.7, 131.4, 128.7 (2C), 128.4, 128.1 (2C), 126.7, 116.4, 62.5, 53.2 (2C), 36.8, 35.8, 32.9, 31.8 (2C).
5-Bromo-N-(3-clorofenll)tlofen-2-carboxamlda (4) 5-Bromo-N-(3-chlorophenyl)tlofen-2-carboxamilde (4)
Ácido 5-bromo-2-tiofencarboxílico (1 ,5 mmol, 310 mg), 3-cloroanilina (1 ,53 mmol, 194 mg), EDC (1 ,5 mmol, 233 mg)E, t3N (3 mmol, 417 pL), HOBt (1 ,5 mmol, 203 mg) y CH2CI2 (6,0 mL). Purificación: Hexano/AcOEt 9:1. Rendimiento: 190 mg, 40%. Sólido blanco. 1H NMR (300 MHz, DMSO-d6) 10 δ,41 (s, 1 H), 7,91 - 7,82 (m, 2H), 7,63 (ddd, J = 8,2, 2,1 , 1 ,0 Hz, 1 H), 7,43 - 7,35 (m, 2H), 7,17 (ddd, J = 8,0, 2,1 , 1 ,0 Hz, 1 H). 13C NMR (75 MHz, DMSO-d6) δ 158,9, 141 ,3, 139,9, 133,0, 131 ,8, 130,4, 130,3, 123,6, 119,7, 118,6, 118,3. 5-Bromo-2-thiophencarboxylic acid (1.5 mmol, 310 mg), 3-chloroaniline (1.53 mmol, 194 mg), EDC (1.5 mmol, 233 mg)E, t3N (3 mmol, 417 pL ), HOBt (1.5 mmol, 203 mg) and CH2CI2 (6.0 mL). Purification: Hexane/AcOEt 9:1. Yield: 190 mg, 40%. White solid. 1 H NMR (300 MHz, DMSO-d6) 10 δ.41 (s, 1 H), 7.91 - 7.82 (m, 2H), 7.63 (ddd, J = 8.2, 2.1 , 1 .0 Hz, 1 H), 7.43 - 7.35 (m, 2H), 7.17 (ddd, J = 8.0, 2.1 , 1 .0 Hz, 1 H). 13 C NMR (75 MHz, DMSO-d6) δ 158.9, 141.3, 139.9, 133.0, 131.8, 130.4, 130.3, 123.6, 119.7, 118, 6, 118.3.
5-Bromo-N-(4-clorofenll)tlofen-2-carboxamlda (5) 5-Bromo-N-(4-chlorophenyl)tlofen-2-carboxamilde (5)
Ácido 5-bromo-2-tiofencarboxílico (3 mmol, 621 mg), 4-cloroanilina (3,06 mmol, 398 mg), EDC (3 mmol, 465 mg)E, t3N (6 mmol, 835 pL), HOBt (3 mmol, 405 mg) y CH2CI2
(10 mL). Purificación: Hexano/AcOEt 7:3. Rendimiento: 368 mg, 39%. Sólido blanco. 1H NMR (300 MHz, DMSO-d6) 1 δ0,39 (s, 1 H), 7,85 (d, J = 4,1 Hz, 1 H), 7,77 - 7,70 (m, 2H), 7,46 - 7,39 (m, 2H), 7,38 (d, J = 4,1 Hz, 1 H). 13C NMR (75 MHz, CDCl3) 6 158,8, 141 ,4, 137,4, 131 ,8, 130,1 , 128,7 (2C), 127,6, 121 ,9 (2C), 118,1. 5-Bromo-2-thiophencarboxylic acid (3 mmol, 621 mg), 4-chloroaniline (3.06 mmol, 398 mg), EDC (3 mmol, 465 mg)E, t3N (6 mmol, 835 pL), HOBt ( 3 mmol, 405 mg) and CH2CI2 (10 mL). Purification: Hexane/AcOEt 7:3. Yield: 368 mg, 39%. White solid. 1 H NMR (300 MHz, DMSO-d6) 1 δ0.39 (s, 1 H), 7.85 (d, J = 4.1 Hz, 1 H), 7.77 - 7.70 (m, 2H ), 7.46 - 7.39 (m, 2H), 7.38 (d, J = 4.1 Hz, 1 H). 13 C NMR (75 MHz, CDCl3) 6 158.8, 141.4, 137.4, 131.8, 130.1, 128.7 (2C), 127.6, 121.9 (2C), 118, 1.
5-Bromo-N-(2-fenlletll)tlofen-2-carboxamlda (6) 5-Bromo-N-(2-fenlletll)tlofen-2-carboxamld (6)
Ácido 5-bromo-2-tiofencarboxílico (3 mmol, 621 mg), feniletilamina (3,06 mmol, 383 pL), EDC (3 mmol, 465 mg), Et3N (6 mmol, 835 pL), HOBt (3 mmol, 405 mg) y CH2CI2 (10 mL). El producto se usó en el paso siguiente sin ninguna purificación posterior. Rendimiento: 253 mg, 56%. Sólido blanco. 1H NMR (300 MHz, DMSO-d6) 8, δ68 (t, J =5-Bromo-2-thiophencarboxylic acid (3 mmol, 621 mg), Phenylethylamine (3.06 mmol, 383 pL), EDC (3 mmol, 465 mg), Et3N (6 mmol, 835 pL), HOBt (3 mmol, 405 mg) and CH2CI2 (10 mL). The product was used in the next step without any further purification. Yield: 253 mg, 56%. White solid. 1 H NMR (300 MHz, DMSO-d6) 8, δ68 (t, J =
5.7 Hz, 1 H), 7,55 (d, J = 4,0 Hz, 1 H), 7,34 - 7,16 (m, 6H), 3,43 (dt, J = 8,0, 6,2 Hz, 2H), 2,81 (dd, J = 8,2, 6,6 Hz, 2H). 13C NMR (75 MHz, DMSO-d6) 159, δ9, 141 ,9, 139,3, 131 ,5,5.7 Hz, 1 H), 7.55 (d, J = 4.0 Hz, 1 H), 7.34 - 7.16 (m, 6H), 3.43 (dt, J = 8.0, 6 ,2 Hz, 2H), 2.81 (dd, J = 8.2, 6.6 Hz, 2H). 13 C NMR (75 MHz, DMSO-d6) 159, δ9, 141 .9, 139.3, 131 .5,
128.7 (2C), 128,5, 128,4 (2C), 126,2, 116,5, 40,8, 35,1. 128.7 (2C), 128.5, 128.4 (2C), 126.2, 116.5, 40.8, 35.1.
5-Bromo-N-(4-morfollnfenll)tlofen-2-carboxamlda (7) 5-Bromo-N-(4-morpholnfenll)tlofen-2-carboxamld (7)
Ácido 5-bromo-2-tiofencarboxílico (3 mmol, 621 mg), anilina (3,06 mmol, 561 mg), EDC (3 mmol, 465 mg)E, t3N (6 mmol, 835 pL), HOBt (3 mmol, 405 mg) y CH2CI2 (10 mL). El producto se usó sin ninguna purificación. Rendimiento: 808 mg, 73%. Sólido blanco. 5-Bromo-2-thiophencarboxylic acid (3 mmol, 621 mg), aniline (3.06 mmol, 561 mg), EDC (3 mmol, 465 mg)E, t3N (6 mmol, 835 pL), HOBt (3 mmol , 405 mg) and CH2CI2 (10 mL). The product was used without any purification. Yield: 808 mg, 73%. White solid.
5-Bromo-N-fenlltlofen-2-carboxamlda (8) 5-Bromo-N-fenlltlofen-2-carboxamld (8)
Ácido 5-bromo-2-tiofencarboxílico (3 mmol, 621 mg), anilina (3,06 mmol, 279 pL), EDC (3 mmol, 465 mg)E, t3N (6 mmol, 835 pL), HOBt (3 mmol, 405 mg) y CH2CI2 (10 mL). Purificación: Hexano/AcOEt 9:1 . Rendimiento: 450 mg, 53 %. Sólido blanco. 1H NMR (300 MHz, DMSO-d6) 1 δ0,28 (s, 1 H), 7,87 (d, J = 4,1 Hz, 1 H), 7,75 - 7,64 (m, 2H), 7,43 - 7,27 (m, 3H), 7,19 - 7,03 (m, 1 H). 13C NMR (75 MHz, DMSO) 6 159,2, 142,3, 138,8, 132,1 , 130,3, 129,1 , 124,4, 120,8, 118,2. 5-Bromo-2-thiophencarboxylic acid (3 mmol, 621 mg), aniline (3.06 mmol, 279 pL), EDC (3 mmol, 465 mg)E, t3N (6 mmol, 835 pL), HOBt (3 mmol , 405 mg) and CH2CI2 (10 mL). Purification: Hexane/AcOEt 9:1. Yield: 450 mg, 53%. White solid. 1 H NMR (300 MHz, DMSO-d6) 1 δ0.28 (s, 1 H), 7.87 (d, J = 4.1 Hz, 1 H), 7.75 - 7.64 (m, 2H ), 7.43 - 7.27 (m, 3H), 7.19 - 7.03 (m, 1H). 13 C NMR (75 MHz, DMSO) 6 159.2, 142.3, 138.8, 132.1, 130.3, 129.1, 124.4, 120.8, 118.2.
Síntesis de 5-(hldroxlfenll)-tlofen-2-carboxamldas (9-20)
Synthesis of 5-(hldroxlfenll)-tlofen-2-carboxymildes (9-20)
Esquema 2. Reactivos y condiciones: ii) Pd(PPh3)4, Na2CO3, tolueno/H2O/EtOH
(2:1 ,5:1), 120 °C (MW). Scheme 2. Reagents and conditions: ii) Pd(PPh 3 )4, Na2CO3, toluene/H 2 O/EtOH (2:1,5:1), 120 °C (MW).
Procedimiento general: sobre una mezcla de la correspondiente 5-bromotiofen-2- carboxamida (1 equiv. obtenida mediante el procedimiento general A), Na2CO3 (2,2 equiv.) y paladio (0) tetrakis(trifenilfosfina) (0,05 equiv.), desgasificada previamente con argón, se añadió una mezcla de tolueno, H2O y EtOH (2:1 , 5:1) sin interrumpir la atmósfera inerte. Finalmente se añadió el correspondiente borano (1 ,2 equiv.) para obtener los derivados sustituidos en orto, meta y para. La mezcla de reacción se burbujeó con argón durante 15 minutos más para su completa desgasificación y a continuación se calentó bajo irradiación de microondas (MW) durante 20 minutos a 120ºC . Finalizado este tiempo, el crudo de reacción se diluyó con una disolución 1 :1 de AcOEt/ H2O y se extrajo con AcOEt. La fase orgánica se lavó con una disolución 1 :1 de disolución saturada de NaCI y H2O, y finalmente se secó sobre Na2SO4 anhidro. A continuación, el desecante se filtró y los volátiles se evaporaron hasta sequedad a vacío. El residuo resultante se purificó por cromatografía en columna utilizando como eluyente mezclas de disolventes, indica en cada caso, para obtener los productos deseados. General procedure: on a mixture of the corresponding 5-bromothiophen-2-carboxamide (1 equiv. obtained by general procedure A), Na2CO3 (2.2 equiv.) and palladium (0) tetrakis (triphenylphosphine) (0.05 equiv. .), previously degassed with argon, a mixture of toluene, H2O and EtOH (2:1, 5:1) was added without interrupting the inert atmosphere. Finally, the corresponding borane (1.2 equiv.) was added to obtain the ortho, meta and para substituted derivatives. The reaction mixture was bubbled with argon for a further 15 minutes for complete degassing and then heated under microwave irradiation (MW) for 20 minutes at 120 ° C. At the end of this time, the reaction crude was diluted with a 1:1 solution of AcOEt/H 2 O and extracted with AcOEt. The organic phase was washed with a 1:1 solution of saturated NaCl solution and H2O, and finally dried over anhydrous Na2SO4. The desiccant was then filtered and the volatiles evaporated to dryness under vacuum. The resulting residue was purified by column chromatography using solvent mixtures as eluent, indicated in each case, to obtain the desired products.
N-(2-(1-Bencllplperldln-4-ll)etll)-5-(2-hldroxlfenll)tlofen-2-carboxamlda (9)N-(2-(1-Bencllplperldln-4-ll)etll)-5-(2-hldroxlfenll)tlofen-2-carboxamlda (9)
Reactivos: N-(2-(1 -bencilpiperidin-4-il)etil)-5-bromotiofen-2-carboxamida (3) (1 ,47 mmol, 601 mg), Na2CO3 (3,23 mmol, 343 mg), paladio (0) tetrakis(trifenilfosfina) (0,074 mmol, 87 mg), 2-(4,4,5,5-tetrametil-1 ,3,2-dioxaborolan-2-il)fenol (1 ,77 mmol, 371 pL) y una solución de tolueno (4 mL), H2O (3 mL) y EtOH (2 mL). Purificación: CH2C/IM2 eOH (95:5). Rendimiento: 116 mg, 20%. Sólido blanco. Pf: 185,5 - 187,5 °C. 1H RMN (300 MHz, DMSO-d6) 1 δ0,42 (s, 1 H), 8,37 (t, J = 5,6 Hz, 1 H), 7,73 - 7,64 (m, 2H), 7,58 (d, J = 4,0 Hz, 1 H), 7,35 - 7,20 (m, 5H), 7,16 (td, J = 7,7, 7,2, 1 ,6 Hz, 1 H), 6,97 (dd, J = 8,2, 1 ,2 Hz, 1 H), 6,87 (td, J = 7,5, 1 ,2 Hz, 1 H), 3,45 (s, 2H), 3,29 - 3,21 (m, 2H), 2,89 - 2,69 (m, 2H.) 2,03 - 1 ,81 (m, 2H), 1 ,73 - 1 ,61 (m, 2H), 1 ,52 - 1 ,38 (m, 2H), 1 ,37 - 1 ,24 (m, 1 H), 1 ,23 - 1 ,08 (m, 2H). 13C RMN (75 MHz, DMSO-d6) 1 δ61 ,3, 153,7, 143,2, 138,5, 129,1 , 128,8, 128,1 (2C), 127,6, 127,4, 126,9, 124,7, 120,0, 119,5, 116,3, 62,4, 53,2 (2C), 36,7, 36,0, 32,9, 31 ,8 (2C). HRMS (ESI) m/z: [M + H]+ cale, para C25H29N2O2S 421 ,1944; encontrado 421 ,1946. Pureza (HPLC) > 99%. Reagents: N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-bromothiophen-2-carboxamide (3) (1.47 mmol, 601 mg), Na2CO3 (3.23 mmol, 343 mg), palladium (0) tetrakis(triphenylphosphine) (0.074 mmol, 87 mg), 2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (1.77 mmol, 371 pL ) and a solution of toluene (4 mL), H2O (3 mL) and EtOH (2 mL). Purification: CH2C/IM2 eOH (95:5). Yield: 116 mg, 20%. White solid. mp: 185.5 - 187.5 °C. 1 H NMR (300 MHz, DMSO-d6) 1 δ0.42 (s, 1 H), 8.37 (t, J = 5.6 Hz, 1 H), 7.73 - 7.64 (m, 2H ), 7.58 (d, J = 4.0 Hz, 1 H), 7.35 - 7.20 (m, 5H), 7.16 (td, J = 7.7, 7.2, 1 , 6 Hz, 1 H), 6.97 (dd, J = 8.2, 1 .2 Hz, 1 H), 6.87 (td, J = 7.5, 1 .2 Hz, 1 H), 3 .45 (s, 2H), 3.29 - 3.21 (m, 2H), 2.89 - 2.69 (m, 2H.) 2.03 - 1 .81 (m, 2H), 1 .73 - 1 .61 (m, 2H), 1 .52 - 1 .38 (m, 2H), 1 .37 - 1 .24 (m, 1 H), 1 .23 - 1 .08 (m, 2H). 13 C NMR (75 MHz, DMSO-d6) 1 δ61 .3, 153.7, 143.2, 138.5, 129.1 , 128.8, 128.1 (2C), 127.6, 127.4 , 126.9, 124.7, 120.0, 119.5, 116.3, 62.4, 53.2 (2C), 36.7, 36.0, 32.9, 31.8 (2C) . HRMS (ESI) m/z: [M + H] + cale, for C25H29N2O2S 421 .1944; found 421 ,1946. Purity (HPLC) > 99%.
N-(2-(1 -Bencllplperldln-4-ll)etll)-5-(3-hldroxlfenll)tlofen-2-carboxamlda (10)N-(2-(1 -Bencllplperldln-4-ll)etll)-5-(3-hldroxlfenll)tlofen-2-carboxamlda (10)
Reactivos: N-(2-(1-bencilpiperidin-4-il)etil)-5-bromotiofen-2-carboxamida (3) (2,21 mmol, 900 mg), Na2CO3 (4,86 mmol, 515 mg), paladio (0) tetrakis(trifenilfosfina) (0,11
mmol, 127 mg), 3-(4,4,5,5-tetrametil-1 ,3,2-dioxaborolan-2-il)fenol (2,65 mmol, 583 mg) y una solución de tolueno (4 mL), H2O (3 mL) y EtOH (2 mL). Purificación: CH2CI2/MeOH (95:5). Rendimiento: 754 mg, 80%. Sólido blanco. Pf: 159 - 161 °C. 1H RMN (300 MHz, DMSO-d6) δ 9,68 (s, 1 H), 8,46 (t, J = 5,6 Hz, 1 H), 7,69 (d, J = 3,9 Hz, 1 H), 7,43 (d, J = 3,9 Hz, 1 H), 7,37 - 7,22 (m, 6H), 7,11 (d, J = 8,0 Hz, 1 H), 7,04 (m, 1 H), 6,76 (dd, J = 7,9, 2,2 Hz, 1 H), 3,43 (s, 2H), 3,26 (m, 2H), 2,78 (m, 2H), 2,03 - 1 ,82 (m, 2H), 1 ,72 - 1 ,60 (m, 2H), 1 ,50 - 1 ,39 (m, 2H), 1 ,34 - 1 ,24 (m, 1 H), 1 ,23 - 1 ,08 (m, 2H). 13C RMN (75 MHz, DMSO-d6) 1 δ60,8, 157,9, 147,3, 138,9, 134,3, 130,3, 128,8, 128,1 (5C), 126,8, 124,0, 116,5, 115,6, 112,3, 62,5, 53,2 (2C), 36,8, 35,9, 32,9, 31 ,8 (2C). HRMS (ESI) m/z: [M + H]+ calc, para C25H29N2O2S 421 ,1944; encontrado 421 ,1943. Pureza (HPLC): 96%. Reagents: N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-bromothiophen-2-carboxamide (3) (2.21 mmol, 900 mg), Na2CO3 (4.86 mmol, 515 mg), palladium (0) tetrakis(triphenylphosphine) (0.11 mmol, 127 mg), 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (2.65 mmol, 583 mg) and a toluene solution (4 mL) , H 2 O (3 mL) and EtOH (2 mL). Purification: CH2CI2/MeOH (95:5). Yield: 754 mg, 80%. White solid. mp: 159 - 161 °C. 1 H NMR (300 MHz, DMSO-d6) δ 9.68 (s, 1 H), 8.46 (t, J = 5.6 Hz, 1 H), 7.69 (d, J = 3.9 Hz, 1H), 7.43 (d, J = 3.9 Hz, 1H), 7.37 - 7.22 (m, 6H), 7.11 (d, J = 8.0 Hz, 1 H), 7.04 (m, 1H), 6.76 (dd, J = 7.9, 2.2 Hz, 1H), 3.43 (s, 2H), 3.26 (m, 2H ), 2.78 (m, 2H), 2.03 - 1.82 (m, 2H), 1.72 - 1.60 (m, 2H), 1.50 - 1.39 (m, 2H), 1 .34 - 1 .24 (m, 1H), 1 .23 - 1 .08 (m, 2H). 13 C NMR (75 MHz, DMSO-d6) 1 δ60.8, 157.9, 147.3, 138.9, 134.3, 130.3, 128.8, 128.1 (5C), 126.8 , 124.0, 116.5, 115.6, 112.3, 62.5, 53.2 (2C), 36.8, 35.9, 32.9, 31.8 (2C). HRMS (ESI) m/z: [M + H] + calc, for C25H29N2O2S 421 .1944; found 421 ,1943. Purity (HPLC): 96%.
N-(2-(1-Bencllplperldln-4-ll)etll)-5-(4-hldroxlfenll)tlofen-2-carboxamlda (11)N-(2-(1-Bencllplperldln-4-ll)etll)-5-(4-hldroxlfenll)tlofen-2-carboxamlda (11)
Reactivos: N-(2-(1-bencilpiperidin-4-il)etil)-5-bromotiofen-2-carboxamida (3) (2,21 mmol, 900 mg), Na2CO3 (4,86 mmol, 515 mg), paladio (0) tetrakis(trifenilfosfina) (0,11 mmol, 127 mg), 4-(4,4,5,5-tetrametil-1 ,3,2-dioxaborolan-2-il)fenol (2,65 mmol, 583 mg) y una solución de tolueno (4 mL), H2O (3 mL) y EtOH (2 mL). Purificación: CH2CI2/MeOH (95:5). Rendimiento: 550 mg, 60%. Sólido blanco. Pf: 164 - 166 °C. 1H RMN (300 MHz, DMSO-d6) δ 9,77 (s, 1 H), 8,37 (t, J = 5,4 Hz, 1 H), 7,66 (d, J = 3,9 Hz, 1 H), 7,50 (d, J = 8,6 Hz, 2H), 7,35 - 7,20 (m, 6H), 6,82 (d, J = 8,6 Hz, 2H), 3,44 (s, 2H), 3,25 (m, 2H), 2,78 (m, 2H), 2,03 - 1 ,82 (m, 2H), 1 ,72 - 1 ,60 (m, 2H), 1 ,51 - 1 ,38 (m, 2H), 1 ,36 - 1 ,24 (m, 1 H), 1 ,23 - 1 ,08 (m, 2H). 13C RMN (75 MHz, DMSO-d6) 160,9 δ, 157,9, 147,9, 137,4, 128,8, 128,1 (5C), 127,1 (2C), 126,9, 124,3, 122,3, 115,9 (2C), 62,4, 53,2 (2C), 36,8, 35,9, 32,9, 31 ,8 (2C). HRMS (ESI) m/z: [M + H]+ calc, para C25H29N2O2S 421 ,1944; encontrado 421 ,1948. Pureza (HPLC): 98%. Reagents: N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-bromothiophen-2-carboxamide (3) (2.21 mmol, 900 mg), Na2CO3 (4.86 mmol, 515 mg), palladium (0) tetrakis(triphenylphosphine) (0.11 mmol, 127 mg), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (2.65 mmol, 583 mg) and a solution of toluene (4 mL), H 2 O (3 mL), and EtOH (2 mL). Purification: CH2CI2/MeOH (95:5). Yield: 550 mg, 60%. White solid. mp: 164 - 166 °C. 1 H NMR (300 MHz, DMSO-d6) δ 9.77 (s, 1 H), 8.37 (t, J = 5.4 Hz, 1 H), 7.66 (d, J = 3.9 Hz, 1H), 7.50 (d, J = 8.6 Hz, 2H), 7.35 - 7.20 (m, 6H), 6.82 (d, J = 8.6 Hz, 2H) , 3.44 (s, 2H), 3.25 (m, 2H), 2.78 (m, 2H), 2.03 - 1 .82 (m, 2H), 1 .72 - 1 .60 (m , 2H), 1 .51 - 1 .38 (m, 2H), 1 .36 - 1 .24 (m, 1 H), 1 .23 - 1 .08 (m, 2H). 13 C NMR (75 MHz, DMSO-d6) 160.9 δ, 157.9, 147.9, 137.4, 128.8, 128.1 (5C), 127.1 (2C), 126.9, 124.3, 122.3, 115.9 (2C), 62.4, 53.2 (2C), 36.8, 35.9, 32.9, 31.8 (2C). HRMS (ESI) m/z: [M + H] + calc, for C25H29N2O2S 421 .1944; found 421 ,1948. Purity (HPLC): 98%.
N-(3-Clorofenll)-5-(2-hldroxlfenll)tlofen-2-carboxamlda (12) N-(3-Chlorofenll)-5-(2-hldroxlfenll)tlofen-2-carboxamld (12)
Reactivos: 5-bromo-N-(3-dorofenil)-2-tiofencarboxamida (4) (0,84 mmol, 265 mg), Na2CO3 (1 ,84 mmol, 195 mg), paladio (0) tetrakis(trifenilfosfina) (0,042 mmol, 49 mg), 2- (4,4,5,5-tetrametil-1 ,3,2-dioxaborolan-2-il)fenol (1 ,00 mmol, 211 pL) y una solución de tolueno (2 mL), H2O (2,5 mL) y EtOH (1 mL). Purificación: Hexano/AcOEt 9:1. Rendimiento: 121 mg, 44%. Sólido blanco. Pf: 246 -248 °C. 1H RMN (300 MHz, DMSO- d6) δ 10,56 (s, 1 H), 10,31 (s, 1 H), 8,00 (d, J = 4,1 Hz, 1 H), 7,94 (t, J = 2,0 Hz, 1 H,), 7,77 (dd, J = 7,9, 1 ,7 Hz, 1 H), 7,73 - 7,67 (m, 2H), 7,39 (t, J = 8,1 Hz, 1 H), 7,24 - 7,13 (m, 2H), 7,01 (dd, J = 8,2, 1 ,2 Hz, 1 H), 6,91 (ddd, J = 7,8, 7,2, 1 ,2 Hz, 1 H). 13C RMN (75 MHz, DMSO-d6) 1 δ60,5, 153,8, 145,0, 140,4, 137,5, 132,9, 130,4 (2C), 129,5, 129,2,
127,6, 124,8, 123,2, 119,8, 119,6, 118,5, 116,4. HRMS (ESI) m/z: [M + H]+ calc, para C17H13CINO2S 330,0350; encontrado 330,0348. Pureza (HPLC): 96,0%. Reagents: 5-bromo-N-(3-dorophenyl)-2-thiophencarboxamide (4) (0.84 mmol, 265 mg), Na2CO3 (1.84 mmol, 195 mg), palladium (0) tetrakis(triphenylphosphine) ( 0.042 mmol, 49 mg), 2- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (1.00 mmol, 211 pL) and a toluene solution (2 mL ), H 2 O (2.5 mL) and EtOH (1 mL). Purification: Hexane/AcOEt 9:1. Yield: 121 mg, 44%. White solid. mp: 246 -248 °C. 1 H NMR (300 MHz, DMSO- d6) δ 10.56 (s, 1 H), 10.31 (s, 1 H), 8.00 (d, J = 4.1 Hz, 1 H), 7 .94 (t, J = 2.0 Hz, 1 H,), 7.77 (dd, J = 7.9, 1.7 Hz, 1 H), 7.73 - 7.67 (m, 2H) , 7.39 (t, J = 8.1 Hz, 1 H), 7.24 - 7.13 (m, 2H), 7.01 (dd, J = 8.2, 1 .2 Hz, 1 H ), 6.91 (ddd, J = 7.8, 7.2, 1 .2 Hz, 1 H). 13 C NMR (75 MHz, DMSO-d6) 1 δ60.5, 153.8, 145.0, 140.4, 137.5, 132.9, 130.4 (2C), 129.5, 129.2 , 127.6, 124.8, 123.2, 119.8, 119.6, 118.5, 116.4. HRMS (ESI) m/z: [M + H] + calc, for C17H13CINO2S 330.0350; found 330.0348. Purity (HPLC): 96.0%.
N-(3-Clorofenll)-5-(3-hldroxlfenll)tlofen-2-carboxamlda (13) N-(3-Chlorofenll)-5-(3-hldroxlfenll)tlofen-2-carboxamld (13)
Reactivos: 5-bromo-N-(3-dorofenil)tiofen-2-carboxamida (4) (0,32 mmol, 101 mg), Na2CO3 (0,71 mmol, 75 mg), paladio (0) tetrakis(trifenilfosfina) (0,016 mmol, 19 mg), 3- (4,4,5,5-tetrametil-1 ,3,2-dioxaborolan-2-il)fenol (0,98 mmol, 85 mg) y una solución de tolueno (2 mL), H2O (1 ,5 mL) y EtOH (1 mL). Purificación: Hexano/AcOEt 1 :1. Rendimiento: 68 mg, 64%. Sólido blanco. Pf: 190 - 192 °C. 1H RMN (300 MHz, DMSO- d6) δ 10,37 (s, 1 H), 9,70 (s, 1 H), 8,01 (d, J = 3,9 Hz, 1 H), 7,92 (t, J = 2,0 Hz, 1 H), 7,72 - 7,63 (m, 1 H), 7,55 (d, J = 4,0 Hz, 1 H), 7,39 (t, J = 8,1 Hz, 1 H), 7,26 (t, 1 H), 7,21 - 7,14 (m, 2H), 7,13 - 7,08 (m, 1 H), 6,83 - 6,77 (m, 1 H). 13C RMN (75 MHz, DMSO) 6 159,9, 157,9, 149,0, 140,2, 137,9, 134,0, 132,9, 130,6, 130,4, 130,4, 124,3, 123,4, 119,6, 118,6, 116,6, 115,9, 112,4. HRMS (ESI) m/z: [M + H]+ calc, para Ci7Hi3CINO2S 330,0350; encontrado 330,0347. Pureza (HPLC) > 99%. Reagents: 5-bromo-N-(3-dorophenyl)thiophen-2-carboxamide (4) (0.32 mmol, 101 mg), Na2CO3 (0.71 mmol, 75 mg), palladium (0) tetrakis(triphenylphosphine) (0.016 mmol, 19 mg), 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (0.98 mmol, 85 mg) and a toluene solution (2 mL), H 2 O (1.5 mL) and EtOH (1 mL). Purification: Hexane/AcOEt 1:1. Yield: 68 mg, 64%. White solid. mp: 190 - 192 °C. 1 H NMR (300 MHz, DMSO- d6) δ 10.37 (s, 1 H), 9.70 (s, 1 H), 8.01 (d, J = 3.9 Hz, 1 H), 7 .92 (t, J = 2.0 Hz, 1 H), 7.72 - 7.63 (m, 1 H), 7.55 (d, J = 4.0 Hz, 1 H), 7.39 (t, J = 8.1 Hz, 1 H), 7.26 (t, 1 H), 7.21 - 7.14 (m, 2H), 7.13 - 7.08 (m, 1 H) , 6.83 - 6.77 (m, 1 H). 13 C NMR (75 MHz, DMSO) 6 159.9, 157.9, 149.0, 140.2, 137.9, 134.0, 132.9, 130.6, 130.4, 130.4, 124.3, 123.4, 119.6, 118.6, 116.6, 115.9, 112.4. HRMS (ESI) m/z: [M + H] + calc, for Ci 7 Hi 3 CINO2S 330.0350; found 330.0347. Purity (HPLC) > 99%.
N-(3-Clorofenll)-5-(4-hldroxlfenll)tlofen-2-carboxamlda (14) N-(3-Chlorofenll)-5-(4-hldroxlfenll)tlofen-2-carboxamld (14)
Reactivos: 5-bromo-N-(3-dorofenil)tiofen-2-carboxamida (4) (1 ,27 mmol, 400 mg), Na2CO3 (2,78 mmol, 295 mg), paladio (0) tetrakis(trifenilfosfina) (0,065 mmol, 75 mg), 4- (4,4,5,5-tetrametil-1 ,3,2-dioxaborolan-2-il)fenol (1 ,52 mmol, 334 mg) y una solución de tolueno (3,1 mL), H2O (2,33 mL) y EtOH (1 ,57 mL). Purificación: Hexano/AcOEt 9:1. Rendimiento: 73 mg, 18%. Sólido amarillo. Pf: descompone. 1H RMN (300 MHz, DMSO- d6) δ 10,31 (s, 1 H), 9,84 (s, 1 H), 7,97 (d, J = 4,0 Hz, 1 H), 7,91 (t, J = 2,0 Hz, 1 H), 7,67 (ddd, J = 8,3, 2,1 , 1 ,0 Hz, 1 H), 7,61 - 7,54 (m, 2H), 7,43 (d, J = 4,0 Hz, 1 H), 7,38 (t, J = 8,1 Hz, 1 H), 7,15 (ddd, J = 8,0, 2,1 , 0,9 Hz, 1 H), 6,88 - 6,81 (m, 2H). 13C RMN (75 MHz, DMSO-d6) δ 159,9, 158,2, 149,8, 140,3, 136,4, 132,9, 130,7, 130,4, 127,3 (2C), 126,9, 124,0, 123,3, 122,6, 119,6, 118,5 (2C). HRMS (ESI) m/z: [M + H]+ calc, para C17H13CINO2S 330,0350; encontrado 330,0346. Pureza (HPLC) > 99%. Reagents: 5-bromo-N-(3-dorophenyl)thiophen-2-carboxamide (4) (1.27 mmol, 400 mg), Na2CO3 (2.78 mmol, 295 mg), palladium (0) tetrakis(triphenylphosphine) (0.065 mmol, 75 mg), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (1.52 mmol, 334 mg) and a toluene solution (3 .1 mL), H 2 O (2.33 mL), and EtOH (1.57 mL). Purification: Hexane/AcOEt 9:1. Yield: 73 mg, 18%. Solid yellow. Pf: decompose. 1 H NMR (300 MHz, DMSO- d6) δ 10.31 (s, 1 H), 9.84 (s, 1 H), 7.97 (d, J = 4.0 Hz, 1 H), 7 .91 (t, J = 2.0 Hz, 1 H), 7.67 (ddd, J = 8.3, 2.1, 1.0 Hz, 1 H), 7.61 - 7.54 (m , 2H), 7.43 (d, J = 4.0 Hz, 1 H), 7.38 (t, J = 8.1 Hz, 1 H), 7.15 (ddd, J = 8.0, 2.1, 0.9 Hz, 1H), 6.88 - 6.81 (m, 2H). 13 C NMR (75 MHz, DMSO-d6) δ 159.9, 158.2, 149.8, 140.3, 136.4, 132.9, 130.7, 130.4, 127.3 (2C) , 126.9, 124.0, 123.3, 122.6, 119.6, 118.5 (2C). HRMS (ESI) m/z: [M + H] + calc, for C17H13CINO2S 330.0350; found 330.0346. Purity (HPLC) > 99%.
N-(4-Clorofenll)-5-(2-hldroxlfenll)tlofen-2-carboxamlda (15) N-(4-Chlorofenll)-5-(2-hldroxlfenll)tlofen-2-carboxamld (15)
Reactivos: 5-bromo-N-(4-dorofenil)tiofen-2-carboxamida (5) (1 ,16 mmol, 368 mg), Na2CO3 (2,55 mmol, 270 mg), paladio (0) tetrakis(trifenilfosfina) (0,058 mmol, 68 mg), 2- (4,4,5,5-tetrametil-1 ,3,2-dioxaborolan-2-il)fenol (1 ,4 mmol, 293 pL) y una soludón de tolueno (2 mL), H2O (1 ,5 mL) y EtOH (1 mL). Purificación: Hexano/AcOEt 9:1. Rendimiento: 100 mg, 26%. Sólido amarillo. Pf: 272 - 274 °C. 1H RMN (300 MHz,
DMSO-d6) δ 10,56 (s, 1 H), 10,29 (s, 1 H), 7,98 (d, J = 4,1 Hz, 1 H), 7,82 - 7,73 (m, 3H), 7,70 (d, J = 4,1 Hz, 1 H), 7,46 - 7,37 (m, 2H), 7,20 (ddd, J = 8,3, 7,3, 1 ,6 Hz, 1 H), 6,99 (dd, J = 8,2, 1 ,2 Hz, 1 H), 6,90 (ddd, J = 8,3, 7,2, 1 ,2 Hz, 1 H). 13C RMN (75 MHz, DMSO-d6) δ 160,3, 153,8, 144,8, 137,9, 137,7, 129,4, 129,0, 128,5 (2C), 127,5, 127,1 , 124,8, 121 ,7 (2C), 119,8, 119,60, 116,4. HRMS (ESI) m/z: [M + H]+ calc, para C17H13CINO2S 330,0350; encontrado 330,0335. Pureza (HPLC): 98,3%. Reagents: 5-bromo-N-(4-dorophenyl)thiophen-2-carboxamide (5) (1.16 mmol, 368 mg), Na2CO3 (2.55 mmol, 270 mg), palladium (0) tetrakis(triphenylphosphine) (0.058 mmol, 68 mg), 2- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (1.4 mmol, 293 pL) and a toluene solution (2 mL), H 2 O (1.5 mL) and EtOH (1 mL). Purification: Hexane/AcOEt 9:1. Yield: 100 mg, 26%. Yellow solid. mp: 272 - 274 °C. 1 H NMR (300 MHz, DMSO-d6) δ 10.56 (s, 1 H), 10.29 (s, 1 H), 7.98 (d, J = 4.1 Hz, 1 H), 7.82 - 7.73 ( m, 3H), 7.70 (d, J = 4.1 Hz, 1 H), 7.46 - 7.37 (m, 2H), 7.20 (ddd, J = 8.3, 7.3 , 1 .6 Hz, 1 H), 6.99 (dd, J = 8.2, 1 .2 Hz, 1 H), 6.90 (ddd, J = 8.3, 7.2, 1 .2 Hz, 1H). 13 C NMR (75 MHz, DMSO-d6) δ 160.3, 153.8, 144.8, 137.9, 137.7, 129.4, 129.0, 128.5 (2C), 127.5 , 127.1 , 124.8, 121 .7 (2C), 119.8, 119.60, 116.4. HRMS (ESI) m/z: [M + H] + calc, for C17H13CINO2S 330.0350; found 330.0335. Purity (HPLC): 98.3%.
N-(4-Clorofenll)-5-(3-hldroxlfenll)tlofen-2-carboxamlda (16) N-(4-Chlorofenll)-5-(3-hldroxlfenll)tlofen-2-carboxamld (16)
Reactivos: 5-bromo-N-(4-dorofenil)tiofen-2-carboxamida (5) (1 ,28 mmol, 404 mg), Na2CO3 (2,82 mmol, 298 mg), paladio (0) tetrakis(trifenilfosfina) (0,065 mmol, 75 mg), 3- (4,4,5,5-tetrametil-1 ,3,2-dioxaborolan-2-il)fenol (1 ,54 mmol, 338 mg) y una solución de tolueno (3,1 mL), H2O (2,33 mL) y EtOH (1 ,57 mL). Purificación: Hexano/AcOEt 9:1. Rendimiento: 180 mg, 43%. Sólido amarillo. Pf: descompone. 1H RMN (300 MHz, DMSO-d6) δ 10,35 (s, 1 H), 9,71 (s, 1 H), 8,00 (d, J = 4,0 Hz, 1 H), 7,83 - 7,73 (m, 2H), 7,55 (d, J = 3,9 Hz, 1 H), 7,48 - 7,38 (m, 2H), 7,26 (t, J = 7,8 Hz), 7,17 (dt, J = 7,7, 1 ,3 Hz, 1 H), 7,10 (t, J = 2,1 Hz, 1 H), 6,80 (ddd, J = 8,0, 2,4, 1 ,1 Hz, 1 H). 13C RMN (75 MHz, DMSO) 6 159,7, 157,9, 148,8, 138,2, 137,7, 134,1 , 130,4, 128,6 (2C), 127,4, 124,3, 121 ,8 (2C), 116,6, 115,9, 112,4. HRMS (ESI) m/z: [M + H]+ calc, para C17H13CINO2S 330,0350; encontrado 330,0355. Pureza (HPLC) > 99%. Reagents: 5-bromo-N-(4-dorophenyl)thiophen-2-carboxamide (5) (1.28 mmol, 404 mg), Na2CO3 (2.82 mmol, 298 mg), palladium (0) tetrakis(triphenylphosphine) (0.065 mmol, 75 mg), 3- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (1.54 mmol, 338 mg) and a toluene solution (3 .1 mL), H2O (2.33 mL) and EtOH (1.57 mL). Purification: Hexane/AcOEt 9:1. Yield: 180 mg, 43%. Yellow solid. Pf: decompose. 1 H NMR (300 MHz, DMSO-d6) δ 10.35 (s, 1 H), 9.71 (s, 1 H), 8.00 (d, J = 4.0 Hz, 1 H), 7 .83 - 7.73 (m, 2H), 7.55 (d, J = 3.9 Hz, 1 H), 7.48 - 7.38 (m, 2H), 7.26 (t, J = 7.8 Hz), 7.17 (dt, J = 7.7, 1.3 Hz, 1 H), 7.10 (t, J = 2.1 Hz, 1 H), 6.80 (ddd, J = 8.0, 2.4, 1 .1 Hz, 1 H). 13 C NMR (75 MHz, DMSO) 6 159.7, 157.9, 148.8, 138.2, 137.7, 134.1, 130.4, 128.6 (2C), 127.4, 124 .3, 121 .8 (2C), 116.6, 115.9, 112.4. HRMS (ESI) m/z: [M + H] + calc, for C17H13CINO2S 330.0350; found 330.0355. Purity (HPLC) > 99%.
N-(4-Clorofenll)-5-(4-hldroxlfenll)tlofen-2-carboxamlda (17) N-(4-Chlorofenll)-5-(4-hldroxlfenll)tlofen-2-carboxamld (17)
Reactivos: 5-bromo-N-(4-dorofenil)tiofen-2-carboxamida (5) (1 ,58 mmol, 500 mg), Na2CO3 (3,48 mmol, 368 mg), paladio (0) tetrakis(trifenilfosfina) (0,081 mmol, 93 mg), 4- (4,4,5,5-tetrametil-1 ,3,2-dioxaborolan-2-il)fenol (1 ,89 mmol, 417 mg) y una solución de tolueno (4 mL), H2O (3 mL) y EtOH (2 mL). Purificación: Hexano/AcOEt 9:1. Rendimiento: 208 mg, 40%. Sólido amarillo. Pf: 291 - 293 °C. 1H RMN (300 MHz, DMSO-d6) δ 10,28 (s, 1 H), 9,83 (s, 1 H), 7,96 (d, J = 4,0 Hz, 1 H), 7,81 - 7,72 (m, 2H), 7,62 - 7,52 (m, 2H), 7,46 - 7,37 (m, 3H), 6,88 - 6,78 (m, 2H). 13C RMN (75 MHz, DMSO) 6 159,9, 158,2, 149,5, 137,8, 136,6, 130,5, 128,6 (2C), 127,2 (2C), 127,2, 124,1 , 122,5, 121 ,7 (2C), 115,9 (2C). HRMS (ESI) m/z: [M + H]+ calc, para C17H13CINO2S 330,0350; encontrado 330,0351 . Pureza (HPLC) > 99%. Reagents: 5-bromo-N-(4-dorophenyl)thiophen-2-carboxamide (5) (1.58 mmol, 500 mg), Na2CO3 (3.48 mmol, 368 mg), palladium (0) tetrakis(triphenylphosphine) (0.081 mmol, 93 mg), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (1.89 mmol, 417 mg) and a toluene solution (4 mL), H 2 O (3 mL) and EtOH (2 mL). Purification: Hexane/AcOEt 9:1. Yield: 208 mg, 40%. Yellow solid. mp: 291 - 293 °C. 1 H NMR (300 MHz, DMSO-d6) δ 10.28 (s, 1 H), 9.83 (s, 1 H), 7.96 (d, J = 4.0 Hz, 1 H), 7 .81 - 7.72 (m, 2H), 7.62 - 7.52 (m, 2H), 7.46 - 7.37 (m, 3H), 6.88 - 6.78 (m, 2H) . 13 C NMR (75 MHz, DMSO) 6 159.9, 158.2, 149.5, 137.8, 136.6, 130.5, 128.6 (2C), 127.2 (2C), 127, 2, 124.1, 122.5, 121.7 (2C), 115.9 (2C). HRMS (ESI) m/z: [M + H] + calc, for C17H13CINO2S 330.0350; found 330.0351 . Purity (HPLC) > 99%.
N-Fenetll-5-(2-hldroxlfenll)tlofen-2-carboxamlda (18) N-Fenetll-5-(2-hldroxlfenll)tlofen-2-carboxamld (18)
Reactivos: 5-bromo-N-(2-feniletil)-2-tiofencarboxamida (6) (2,23 mmol, 692 mg), Na2CO3 (4,91 mmol, 520 mg), paladio (0) tetrakis(trifenilfosfina) (0,114 mmol, 131 mg),
2-(4,4,5,5-tetrametil-1 ,3,2-dioxaborolan-2-il)fenol (2,68 mmol, 561 pL) y una solución de tolueno (4 mL), H2O (3 mL) y EtOH (2 mL). El producto se usó sin ninguna purificación. Rendimiento: 263 mg, 45%. Sólido blanco. Pf: 200 -202 °C. 1H RMN (300 MHz, DMSO- d6) δ 10,42 (s, 1 H), 8,53 (t, J = 5,7 Hz, 1 H), 7,72 - 7,64 (m, 2H), 7,58 (d, J = 4,0 Hz, 1 H), 7,34 - 7,13 (m, 6H), 6,97 (dd, J = 8,2, 1 ,2 Hz, 1 H), 6,87 (td, J = 7,5, 1 ,3 Hz, 1 H), 3,46 (dt, J = 8,0, 6,1 Hz, 2H), 2,84 (dd, J = 8,3, 6,6 Hz, 2H). 13C RMN (75 MHz, DMSO-d6) δ 161 ,4, 153,7, 143,4, 139,5, 138,3, 129,1 , 128,6 (2C), 128,3 (2C), 127,6, 126,1 , 124,7, 120,0, 119,5, 116,3, 40,7, 35,2. HRMS (ESI) m/z: [M + H]+ cale, para C19H18NO2S 324,1053; encontrado 324,1043. Pureza (HPLC)> 99%. Reagents: 5-bromo-N-(2-phenylethyl)-2-thiophencarboxamide (6) (2.23 mmol, 692 mg), Na2CO3 (4.91 mmol, 520 mg), palladium (0) tetrakis(triphenylphosphine) ( 0.114 mmol, 131 mg), 2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (2.68 mmol, 561 pL) and a toluene solution (4 mL), H 2 O (3 mL) and EtOH (2 mL). The product was used without any purification. Yield: 263 mg, 45%. White solid. mp: 200 -202 °C. 1 H NMR (300 MHz, DMSO- d6) δ 10.42 (s, 1 H), 8.53 (t, J = 5.7 Hz, 1 H), 7.72 - 7.64 (m, 2H ), 7.58 (d, J = 4.0 Hz, 1 H), 7.34 - 7.13 (m, 6H), 6.97 (dd, J = 8.2, 1 .2 Hz, 1 H), 6.87 (td, J = 7.5, 1.3 Hz, 1 H), 3.46 (dt, J = 8.0, 6.1 Hz, 2H), 2.84 (dd, J = 8.3, 6.6 Hz, 2H). 13 C NMR (75 MHz, DMSO-d6) δ 161.4, 153.7, 143.4, 139.5, 138.3, 129.1, 128.6 (2C), 128.3 (2C), 127.6, 126.1, 124.7, 120.0, 119.5, 116.3, 40.7, 35.2. HRMS (ESI) m/z: [M + H] + cale, for C 19 H 18 NO 2 S 324.1053; found 324.1043. Purity (HPLC) > 99%.
N-Fenetll-5-(3-hldroxlfenll)tlofen-2-carboxamlda (19) N-Fenetll-5-(3-hldroxlfenll)tlofen-2-carboxamld (19)
Reactivos: 5-bromo-N-(2-feniletil)-2-tiofencarboxamida (6) (1 ,13 mmol, 350 mg), Na2CO3 (2,49 mmol, 264 mg), paladio (0) tetrakis(trifenilfosfina) (0,058 mmol, 67 mg), 3- (4,4,5,5-tetrametil-1 ,3,2-dioxaborolan-2-il)fenol (1 ,36 mmol, 298 mg) y una solución de tolueno (2 mL), H2O (1 ,5 mL) y EtOH (1 mL). Purificación: Hexano/AcOEt 1 :1. Rendimiento: 70 mg, 19%. Sólido blanco. Pf: 183 - 185 °C. 1H RMN (300 MHz, DMSO- d6) δ 9,67 (s, 1 H), 8,62 (t, J = 5,6 Hz, 1 H), 7,69 (d, J = 3,9 Hz, 1 H), 7,44 (d, J = 3,9 Hz, 1 H), 7,34 - 7,17 (m, 6H), 7,12 (dt, J = 7,7, 1 ,3 Hz, 1 H), 7,05 (t, J = 2,0 Hz, 1 H), 6,77 (ddd, J = 8,0, 2,4, 1 ,0 Hz, 1 H), 3,46 (dt, J = 8,1 , 6,2 Hz, 2H), 2,84 (dd, J = 8,4, 6,5 Hz, 2H).13C RMN (75 MHz, DMSO) δ 160,9, 157,9, 147,4, 139,4, 138,8, 134,3, 130,3, 128,9, 128,7 (2C), 128,4 (2C), 126,1 , 124,1 , 116,5, 115,6, 112,3, 40,8, 35,2. HRMS (ESI) m/z: [M + H]+ cale, para C19H18NO2S 324,1053; encontrado 324,1049. Pureza (HPLC) > 99%. Reagents: 5-bromo-N-(2-phenylethyl)-2-thiophencarboxamide (6) (1.13 mmol, 350 mg), Na2CO3 (2.49 mmol, 264 mg), palladium (0) tetrakis(triphenylphosphine) ( 0.058 mmol, 67 mg), 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (1.36 mmol, 298 mg) and a toluene solution (2 mL ), H2O (1.5 mL) and EtOH (1 mL). Purification: Hexane/AcOEt 1:1. Yield: 70 mg, 19%. White solid. mp: 183 - 185 °C. 1 H NMR (300 MHz, DMSO- d6) δ 9.67 (s, 1 H), 8.62 (t, J = 5.6 Hz, 1 H), 7.69 (d, J = 3.9 Hz, 1H), 7.44 (d, J = 3.9 Hz, 1H), 7.34 - 7.17 (m, 6H), 7.12 (dt, J = 7.7, 1, 3 Hz, 1 H), 7.05 (t, J = 2.0 Hz, 1 H), 6.77 (ddd, J = 8.0, 2.4, 1 .0 Hz, 1 H), 3 .46 (dt, J = 8.1, 6.2 Hz, 2H), 2.84 (dd, J = 8.4, 6.5 Hz, 2H). 13 C NMR (75 MHz, DMSO) δ 160.9, 157.9, 147.4, 139.4, 138.8, 134.3, 130.3, 128.9, 128.7 (2C), 128 .4 (2C), 126.1, 124.1, 116.5, 115.6, 112.3, 40.8, 35.2. HRMS (ESI) m/z: [M + H] + cale, for C 19 H 18 NO 2 S 324.1053; found 324.1049. Purity (HPLC) > 99%.
5-(2-Hldroxlfenll)-N-(4-morfollnofenll)tlofen-2-carboxamlda (20) 5-(2-Hldroxlfenll)-N-(4-morfollnofenll)tlofen-2-carboxamld (20)
Reactivos: 5-bromo-N-(4-morfolinofenil)tiofen-2-carboxamida (7) (2,2 mmol, 809 mg), Na2CO3 (4,84 mmol, 513 mg), paladio (0) tetrakis(trifenilfosfina) (0,11 mmol, 130 mg), 2- (4,4,5,5-tetrametil-1 ,3,2-dioxaborolan-2-il)fenol (2,64 mmol, 553 pL) y una solución de tolueno (3,1 mL), H2O (2,3 mL) y EtOH (1 ,55 mL). Purificación: filtración del sólido y lavado con agua y AcOEt. Rendimiento: 774 mg, 92%. Sólido amarillo. Pf : 264 - 266 ºC. 1H RMN (300 MHz, DMSO-d6) δ 9,99 (s, 1 H), 7,92 (d, J = 4,1 Hz, 1 H), 7,72 (dd, J = 8,0, 1 ,6 Hz, 1 H), 7,65 (d, J = 4,1 Hz, 1 H), 7,63 - 7,56 (m, 2H), 7,13 (td, J = 7,6, 7,1 , 1 ,6 Hz, 1 H), 7,01 (dd, J = 8,3, 1 ,2 Hz, 1 H), 6,97 - 6,89 (m, 2H), 6,85 - 6,75 (m, 1 H), 3,80 - 3,67 (m, 4H), 3,13 - 3,00 (m, 4H). 13C RMN (75 MHz, DMSO-d6) 160 δ,0, 155,7, 147,3, 144,8, 137,9, 131 ,1 , 129,1 , 128,1 , 127,1 , 123,9, 121 ,4 (2C), 119,9, 118,1 , 116,8, 115,3 (2C), 66,1 (2C), 48,9 (2C). HRMS (ESI) m/z: [M + H]+ cale, para C21H20N2O3S 381 ,1267;
encontrado 381 ,1267. Pureza (HPLC): 97,8%. Reagents: 5-bromo-N-(4-morpholinophenyl)thiophen-2-carboxamide (7) (2.2 mmol, 809 mg), Na2CO3 (4.84 mmol, 513 mg), palladium (0) tetrakis(triphenylphosphine) (0.11 mmol, 130 mg), 2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (2.64 mmol, 553 pL) and a toluene solution (3.1 mL), H 2 O (2.3 mL), and EtOH (1.55 mL). Purification: filtration of the solid and washing with water and AcOEt. Yield: 774 mg, 92%. Yellow solid. mp: 264 - 266 º C. 1 H NMR (300 MHz, DMSO-d6) δ 9.99 (s, 1 H), 7.92 (d, J = 4.1 Hz, 1 H), 7.72 (dd, J = 8.0, 1.6 Hz, 1 H), 7.65 (d, J = 4.1 Hz, 1 H), 7.63 - 7.56 (m, 2H), 7, 13 (td, J = 7.6, 7.1, 1.6 Hz, 1 H), 7.01 (dd, J = 8.3, 1.2 Hz, 1 H), 6.97 - 6, 89 (m, 2H), 6.85 - 6.75 (m, 1H), 3.80 - 3.67 (m, 4H), 3.13 - 3.00 (m, 4H). 13 C NMR (75 MHz, DMSO-d6) 160 δ,0, 155.7, 147.3, 144.8, 137.9, 131,1, 129.1, 128.1, 127.1, 123, 9, 121.4 (2C), 119.9, 118.1, 116.8, 115.3 (2C), 66.1 (2C), 48.9 (2C). HRMS (ESI) m/z: [M + H] + cale, for C21H20N2O3S 381 .1267; found 381 .1267. Purity (HPLC): 97.8%.
Síntesis de (plperldln-4-lloxl)fenll)tlofen-2-carboxamldas (21-30)
Synthesis of (plperldln-4-lloxl)fenll)tlofen-2-carboxymildes (21-30)
Esquema 3. Reactivos y condiciones, (iii) DIAD, PPh3, THF, 0 °C a 25ºC ; (iv) TFA, CH2CI2, 25 °C. Scheme 3. Reagents and conditions, (iii) DIAD, PPh3, THF, 0 °C to 25 º C; (iv) TFA, CH2CI2, 25 °C.
Procedimiento general: a una solución de trifenilfosfina (1 ,3 eq) en THF a 0 °C se añadió DIAD (1 ,3 eq). La mezcla de reacción se agitó a temperatura ambiente hasta que apareció una turbidez blanca. Tras enfriar de nuevo a 0 ºC, se añadió una solución del precursor 5-(2-hidroxifenil)-tiofen-2-carboxamida (1 equiv., obtenido mediante el procedimiento general B) y 4-hidroxipiperidina-1-carboxilato de tere-butilo (1 ,3 equiv.) en THF. Se dejó agitando toda la noche y a continuación se eliminó el disolvente a presión reducida. El residuo sólido blanco obtenido, correspondiente al derivado protegido con Boc, se disolvió en una solución de CH2CI2/TFA 3:2 y se agitó a temperatura ambiente durante 3 horas. Finalizada la desprotección, se evaporó el disolvente a vacío y tras disolver el producto resultante en CH2CI2 se basificó con NaHCO3 para obtener el producto en forma neutra. A continuación, los volátiles se evaporaron hasta sequedad a vacío y el residuo resultante se purificó por cromatografía en columna utilizando como eluyente las mezclas de disolventes, indicadas en cada caso, para obtener los productos deseados. General procedure: DIAD (1.3 eq) was added to a solution of triphenylphosphine (1.3 eq) in THF at 0 °C. The reaction mixture was stirred at room temperature until white turbidity appeared. After cooling again to 0 ° C, a solution of the precursor 5-(2-hydroxyphenyl)-thiophen-2-carboxamide (1 equiv., obtained by general procedure B) and tere 4-hydroxypiperidine-1-carboxylate was added. -butyl (1.3 equiv.) in THF. It was left stirring overnight and then the solvent was removed under reduced pressure. The white solid residue obtained, corresponding to the Boc-protected derivative, was dissolved in a 3:2 CH2CI2/TFA solution and stirred at room temperature for 3 hours. Once the deprotection was completed, the solvent was evaporated under vacuum and after dissolving the resulting product in CH2CI2, it was basified with NaHCO3 to obtain the product in neutral form. The volatiles were then evaporated to dryness in vacuo and the resulting residue was purified by column chromatography using the solvent mixtures indicated in each case as eluent to obtain the desired products.
N-(2-(1-Bencllplperldln-4-ll)etll)-5-(2-(plperldln-4-lloxl)fenll)tlofen-2-carboxamldaN-(2-(1-Bencllplperldln-4-ll)etll)-5-(2-(plperldln-4-lloxl)fenll)tlofen-2-carboxamlda
(21) (twenty-one)
Reactivos: N-(2-(1 -bencilpiperidin-4-il)etil)-5-(2-hidroxifenil)tiofen-2-carboxamida (9) (1 ,66 mmol, 700 mg), 4-hidroxipiperidin-1 -carboxilato de tere-butilo (2,16 mmol, 434 mg), azodicarboxilato de diisopropilo (2,16 mmol, 426 μL), trifenilfosfina (2,16 mmol, 434 mg) y 15 mL de THF. Se obtuvieron 150 mg de 4-(5-((2-(1 -bencilpiperidin-4-
il)etil)carbamoil)tiofen-2-il)fenoxi)piperidin-1-carboxilato de terc-butilo que se trataron con una solución de CH2CI2 (3 mL) y ácido trifluoroacético (2 mL). Purificación: AcOEt/MeOH 8:2. Rendimiento: 30 mg, 24%. Sólido blanco. Pf: 118 - 120 °C. 1H RMN (300 MHz, DMSO-d6) 68,39 (t, J = 5,7 Hz, 1 H), 7,77 (dd, J = 7,9, 1 ,7 Hz, 1 H), 7,68 (d, J = 4,0 Hz, 1 H), 7,58 (d, J = 4,0 Hz, 1 H), 7,33 - 7,26 (m, 5H), 7,26 - 7,18 (m, 2H), 7,02 - 6,97 (m, 1 H), 4,67 - 4,58 (m, 1 H), 3,42 (s, 2H), 3,28 - 3,23 (m, 2H), 3,01 - 2,95 (m, 2H), 2,81 - 2,74 (m, 2H), 2,58 (ddt, 2H), 2,01 - 1 ,94 (m, 2H), 1 ,92 - 1 ,86 (m, 2H), 1 ,70 - 1 ,64 (m, 2H), 1 ,64 - 1 ,55 (m, 2H), 1 ,46 (q, J = 7,0 Hz, 2H), 1 ,35 - 1 ,26 (m, 1 H), 1 ,20 - 1 ,12 (m, 2H). 13C RMN (75 MHz, DMSO-d6) 161 δ ,3, 152,9, 142,6, 139,2, 138,7, 129,2, 128,7 (2C), 128,1 (2C), 128,0, 127,0, 126,7, 125,2, 122,6, 120,8, 114,3, 74,5, 62,5, 53,3 (2C), 43,6 (2C), 36,7, 36,0, 32,9, 32,2 (2C), 31 ,9 (2C). HRMS (ESI) m/z: [M + H]+ cale, para C30H38N3O2S 504,2679; encontrado 504,2680. Pureza (HPLC): 98,7%. Reagents: N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-(2-hydroxyphenyl)thiophen-2-carboxamide (9) (1.66 mmol, 700 mg), 4-hydroxypiperidin-1 - tere-butyl carboxylate (2.16 mmol, 434 mg), diisopropyl azodicarboxylate (2.16 mmol, 426 μL), triphenylphosphine (2.16 mmol, 434 mg), and 15 mL of THF. 150 mg of 4-(5-((2-(1 -benzylpiperidin-4- tert-butyl yl)ethyl)carbamoyl)thiophen-2-yl)phenoxy)piperidine-1-carboxylate which were treated with CH2CI2 solution (3 mL) and trifluoroacetic acid (2 mL). Purification: AcOEt/MeOH 8:2. Yield: 30 mg, 24%. White solid. mp: 118 - 120 °C. 1 H NMR (300 MHz, DMSO-d6) 68.39 (t, J = 5.7 Hz, 1 H), 7.77 (dd, J = 7.9, 1.7 Hz, 1 H), 7 .68 (d, J = 4.0 Hz, 1 H), 7.58 (d, J = 4.0 Hz, 1 H), 7.33 - 7.26 (m, 5H), 7.26 - 7.18 (m, 2H), 7.02 - 6.97 (m, 1H), 4.67 - 4.58 (m, 1H), 3.42 (s, 2H), 3.28 - 3.23 (m, 2H), 3.01 - 2.95 (m, 2H), 2.81 - 2.74 (m, 2H), 2.58 (ddt, 2H), 2.01 - 1, 94 (m, 2H), 1 .92 - 1 .86 (m, 2H), 1 .70 - 1 .64 (m, 2H), 1 .64 - 1 .55 (m, 2H), 1 .46 ( q, J = 7.0 Hz, 2H), 1 .35 - 1 .26 (m, 1 H), 1 .20 - 1 .12 (m, 2H). 13 C NMR (75 MHz, DMSO-d6) 161 δ ,3, 152.9, 142.6, 139.2, 138.7, 129.2, 128.7 (2C), 128.1 (2C), 128.0, 127.0, 126.7, 125.2, 122.6, 120.8, 114.3, 74.5, 62.5, 53.3 (2C), 43.6 (2C), 36.7, 36.0, 32.9, 32.2 (2C), 31.9 (2C). HRMS (ESI) m/z: [M + H] + cale, for C30H38N3O2S 504.2679; found 504.2680. Purity (HPLC): 98.7%.
N-(3-Clorofenll)-5-(2-(plperldln-4-lloxl)fenll)tlofen-2-carboxamlda (22)N-(3-Chlorofenll)-5-(2-(plperldln-4-lloxl)fenll)tlofen-2-carboxamld (22)
Reactivos: N-(3-clorofenil)-5-(2-hidroxifenil)tiofen-2-carboxamida (12) (0,34 mmol, 112 mg), 4-hidroxipiperidin-1-carboxilato de terc-butilo (0,44 mmol, 89 mg), azodicarboxilato de diisopropilo (0,44 mmol, 87 pL), trifenilfosfina (0,44 mmol, 115 mg) y 3 mL de THF. Purificación: Hexano/AcOEt (8:2). Se obtuvieron 169 mg de 4-(5-((3- dorofenil)carbamoil)tiofen-2-il)fenoxi)piperidin-1-carboxilato de terc-butilo que se trataron con una solución de CH2CI2 (3 mL) y ácido trifluoroacético (2 mL). Purificación: CH2Cl2/MeOH 9:1. Rendimiento: 53 mg, 39%. Sólido blanco. Pf: 195 - 197 °C. 1H RMN (500 MHz, DMSO-d6) 1 δ0,42 (s, 1 H), 8,06 (d, J = 4,1 Hz, 1 H), 7,96 (t, J = 2,1 Hz, 1 H), 7,86 (dd, J = 7,9, 1 ,7 Hz, 1 H), 7,73 - 7,68 (m, 2H), 7,38 (t, J = 8,1 Hz, 1 H), 7,33 (ddd, J = 8,7, 7,2, 1 ,7 Hz, 1 H), 7,23 (d, J = 8,6 Hz, 1 H), 7, 15 (ddd, J = 8,0, 2, 1 , 0,9 Hz, 1 H), 7,05 - 7,01 (m, 1 H), 4,72 - 4,66 (m, 1 H), 3,05 - 3,00 (m, 2H), 2,67 - 2,59 (m, 2H), 2,05 - 1 ,97 (m, 2H), 1 ,69 - 1 ,61 (m, 2H).13C RMN (125 MHz, DMSO-d6) 160 δ,5, 152,9, 144,2, 140,4, 138,3, 132,9, 130,3, 129,6, 128,9, 128,1 , 125,4, 123,2, 122,3, 120,9, 119,6, 118,5, 114,3, 74,4, 43,4 (2C), 31 ,9 (2C). HRMS (ESI) m/z: [M + H]+ cale, para C22H22CIN2O2S 413,1085; encontrado 413,1083. Pureza (HPLC) > 99%. Reagents: N-(3-chlorophenyl)-5-(2-hydroxyphenyl)thiophen-2-carboxamide (12) (0.34 mmol, 112 mg), tert-butyl 4-hydroxypiperidine-1-carboxylate (0.44 mmol, 89 mg), diisopropyl azodicarboxylate (0.44 mmol, 87 pL), triphenylphosphine (0.44 mmol, 115 mg) and 3 mL of THF. Purification: Hexane/AcOEt (8:2). 169 mg of tert-butyl 4-(5-((3-dorophenyl)carbamoyl)thiophen-2-yl)phenoxy)piperidine-1-carboxylate were obtained, which were treated with a solution of CH2CI2 (3 mL) and trifluoroacetic acid. (2 mL). Purification: CH 2 Cl2/MeOH 9:1. Yield: 53 mg, 39%. White solid. mp: 195 - 197 °C. 1 H NMR (500 MHz, DMSO-d6) 1 δ0.42 (s, 1 H), 8.06 (d, J = 4.1 Hz, 1 H), 7.96 (t, J = 2.1 Hz, 1 H), 7.86 (dd, J = 7.9, 1 .7 Hz, 1 H), 7.73 - 7.68 (m, 2H), 7.38 (t, J = 8, 1 Hz, 1 H), 7.33 (ddd, J = 8.7, 7.2, 1 .7 Hz, 1 H), 7.23 (d, J = 8.6 Hz, 1 H), 7 , 15 (ddd, J = 8.0, 2, 1 , 0.9 Hz, 1 H), 7.05 - 7.01 (m, 1 H), 4.72 - 4.66 (m, 1 H ), 3.05 - 3.00 (m, 2H), 2.67 - 2.59 (m, 2H), 2.05 - 1 .97 (m, 2H), 1 .69 - 1 .61 (m , 2H). 13 C NMR (125 MHz, DMSO-d6) 160 δ,5, 152.9, 144.2, 140.4, 138.3, 132.9, 130.3, 129.6, 128.9, 128, 1, 125.4, 123.2, 122.3, 120.9, 119.6, 118.5, 114.3, 74.4, 43.4 (2C), 31.9 (2C). HRMS (ESI) m/z: [M + H] + cale, for C22H22CIN2O2S 413.1085; found 413.1083. Purity (HPLC) > 99%.
N-(3-Clorofenll)-5-(3-(plperldln-4-lloxl)fenll)tlofen-2-carboxamlda (23)N-(3-Chlorofenll)-5-(3-(plperldln-4-lloxl)fenll)tlofen-2-carboxamld (23)
Reactivos: N-(3-clorofenil)-5-(3-hidroxifenil)tiofen-2-carboxamida (13) (0,62 mmol, 206 mg), 4-hidroxipiperidin-1-carboxilato de terc-butilo (0,81 mmol, 163 mg), azodicarboxilato de diisopropilo (0,81 mmol, 160 pL), trifenilfosfina (0,81 mmol, 213 mg) y 10 mL de THF. Se obtuvieron 139 mg de 4-(5-((3-clorofenil)carbamoil)tiofen-2-
il)fenoxi)piperidin-1 -carboxilato de terc-butilo que se trataron con una solución de CH2CI2 (3 mL) y ácido trifluoroacético (2 mL). Purificación: CH2CL2/MeOH 9:1. Rendimiento: 17 mg, 16 %. Sólido amarillo. Pf: 150 - 152 °C. 1H RMN (300 MHz, DMSO-d6) 10 δ,40 (s, 1 H), 8,03 (d, J = 4,0 Hz, 1 H), 7,92 (t, J = 2,1 Hz, 1 H), 7,72 - 7,63 (m, 2H), 7,43 - 7,25 (m, 4H), 7,20 - 7,14 (m, 1 H), 7,03 - 6,95 (m, 1 H), 4,62 - 4,45 (m, 1 H), 3,04 - 2,91 (m, 2H), 2,69 - 2,57 (m, 2H), 2,02 - 1 ,90 (m, 2H), 1 ,58 - 1 ,41 (m, 2H), 1 ,27 - 1 ,10 (m, 1 H). 13C RMN (75 MHz, DMSO-d6) 15 δ9,9, 157,6, 148,7, 140,2, 138,3, 134,3, 133,0, 130,6, 130,5, 130,4, 124,8, 123,4, 119,7, 118,6, 118,2, 116,1 , 113,1 , 73,3, 43,5 (2C), 31 ,9 (2C). HRMS (ESI) m/z: [M + H]+ cale, para C22H22CIN2O2S 413,1085; encontrado 413,1080. Pureza (HPLC) > 99%. Reagents: N-(3-chlorophenyl)-5-(3-hydroxyphenyl)thiophen-2-carboxamide (13) (0.62 mmol, 206 mg), tert-butyl 4-hydroxypiperidine-1-carboxylate (0.81 mmol, 163 mg), diisopropyl azodicarboxylate (0.81 mmol, 160 pL), triphenylphosphine (0.81 mmol, 213 mg) and 10 mL of THF. 139 mg of 4-(5-((3-chlorophenyl)carbamoyl)thiophen-2- were obtained. tert-butyl yl)phenoxy)piperidine-1-carboxylate which were treated with a solution of CH2CI2 (3 mL) and trifluoroacetic acid (2 mL). Purification: CH2CL2/MeOH 9:1. Yield: 17 mg, 16%. Solid yellow. mp: 150 - 152 °C. 1 H NMR (300 MHz, DMSO-d6) 10 δ.40 (s, 1 H), 8.03 (d, J = 4.0 Hz, 1 H), 7.92 (t, J = 2.1 Hz, 1H), 7.72 - 7.63 (m, 2H), 7.43 - 7.25 (m, 4H), 7.20 - 7.14 (m, 1H), 7.03 - 6.95 (m, 1H), 4.62 - 4.45 (m, 1H), 3.04 - 2.91 (m, 2H), 2.69 - 2.57 (m, 2H), 2.02 - 1 .90 (m, 2H), 1 .58 - 1 .41 (m, 2H), 1 .27 - 1 .10 (m, 1 H). 13 C NMR (75 MHz, DMSO-d6) 15 δ9.9, 157.6, 148.7, 140.2, 138.3, 134.3, 133.0, 130.6, 130.5, 130, 4, 124.8, 123.4, 119.7, 118.6, 118.2, 116.1, 113.1, 73.3, 43.5 (2C), 31.9 (2C). HRMS (ESI) m/z: [M + H] + cale, for C22H22CIN2O2S 413.1085; found 413.1080. Purity (HPLC) > 99%.
N-(3-clorofenll)-5-(4-(plperldln-4-lloxl)fenll)tlofen-2-carboxamlda (24)N-(3-chlorophenll)-5-(4-(plperldln-4-lloxl)fenll)tlofen-2-carboxamld (24)
Reactivos: N-(3-clorofenil)-5-(4-hidroxifenil)tiofen-2-carboxamida (14) (0,88 mmol, 290 mg), 4-hidroxipiperidin-1 -carboxilato de terc-butilo (1 ,14 mmol, 229 mg), azodicarboxilato de diisopropilo (1 ,14 mmol, 226 pL), trifenilfosfina (1 ,14 mmol, 289 mg) y 5 mL de THF. Se obtuvieron 213 mg de 4-(4-(5-((3-clorofenil)carbamoil)tiofen-2- il)fenoxi)piperidin- 1 -carboxilato de terc-butilo que se trataron con una solución de CH2CI2 (4 mL) y ácido trifluoroacético (1 mL). Purificación: CH2CL2/MeOH 9:1 . Rendimiento: 38 mg, 22%. Sólido blanco. Pf: 224 - 226 °C. 1H RMN (300 MHz, DMSO-d6) 1 δ0,37 (s, 1 H), 8,02 (d, J = 4,0 Hz, 1 H), 7,92 (t, J = 2,0 Hz, 1 H), 7,73 - 7,61 (m, 3H), 7,50 (d, J = 3,9 Hz, 1 H), 7,39 (t, J = 8,1 Hz, 1 H), 7,20 - 7,13 (m, 1 H), 7,09 - 6,99 (m, 2H), 4,58 - 4,40 (m, 1 H), 3,05 - 2,91 (m, 2H), 2,71 - 2,58 (m, 2H), 2,09 - 1 ,87 (m, 2H), 1 ,61 - 1 ,40 (m, 2H). 13C RMN (75 MHz, DMSO) 6 159,9, 157,6, 149,1 , 140,3, 137,0, 132,9, 130,8, 130,4, 127,3 (2C), 125,5, 123,3, 123,2, 119,6, 118,5, 116,4 (2C), 72,9, 43,2 (2C), 31 ,5 (2C). HRMS (ESI) m/z: [M + H]+ cale, para C22H22CIN2O2S 413,1085; encontrado 413,1083. Pureza (HPLC) > 99%. Reagents: N-(3-chlorophenyl)-5-(4-hydroxyphenyl)thiophen-2-carboxamide (14) (0.88 mmol, 290 mg), tert-butyl 4-hydroxypiperidin-1-carboxylate (1,14 mmol, 229 mg), diisopropyl azodicarboxylate (1.14 mmol, 226 pL), triphenylphosphine (1.14 mmol, 289 mg) and 5 mL of THF. 213 mg of tert-butyl 4-(4-(5-((3-chlorophenyl)carbamoyl)thiophen-2-yl)phenoxy)piperidine-1-carboxylate were obtained, which were treated with a CH2CI2 solution (4 mL). and trifluoroacetic acid (1 mL). Purification: CH2CL2/MeOH 9:1. Yield: 38 mg, 22%. White solid. mp: 224 - 226 °C. 1 H NMR (300 MHz, DMSO-d6) 1 δ0.37 (s, 1 H), 8.02 (d, J = 4.0 Hz, 1 H), 7.92 (t, J = 2.0 Hz, 1 H), 7.73 - 7.61 (m, 3H), 7.50 (d, J = 3.9 Hz, 1 H), 7.39 (t, J = 8.1 Hz, 1 H), 7.20 - 7.13 (m, 1 H), 7.09 - 6.99 (m, 2H), 4.58 - 4.40 (m, 1 H), 3.05 - 2, 91 (m, 2H), 2.71 - 2.58 (m, 2H), 2.09 - 1 .87 (m, 2H), 1 .61 - 1 .40 (m, 2H). 13 C NMR (75 MHz, DMSO) 6 159.9, 157.6, 149.1, 140.3, 137.0, 132.9, 130.8, 130.4, 127.3 (2C), 125 .5, 123.3, 123.2, 119.6, 118.5, 116.4 (2C), 72.9, 43.2 (2C), 31 .5 (2C). HRMS (ESI) m/z: [M + H] + cale, for C22H22CIN2O2S 413.1085; found 413.1083. Purity (HPLC) > 99%.
N-(4-Clorofenll)-5-(2-(plperldln-4-lloxl)fenll)tlofen-2-carboxamlda (25)N-(4-Chlorofenll)-5-(2-(plperldln-4-lloxl)fenll)tlofen-2-carboxamld (25)
Reactivos: N-(4-clorofenil)-5-(2-hidroxifenil)tiofen-2-carboxamida (15) (0,76 mmol, 250 mg), 4-hidroxipiperidin-1 -carboxilato de terc-butilo (0,99 mmol, 199 mg), azodicarboxilato de diisopropilo (0,99 mmol, 195 pL), trifenilfosfina (0,99 mmol, 259 mg) y 7 mL de THF. Se obtuvieron 238 mg de 4-(5-((4-clorofenil)carbamoil)tiofen-2- il)fenoxi)piperidin- 1 -carboxilato de terc-butilo (238 mg) que se trataron con una solución de CH2CI2 (5 mL) y ácido trifluoroacético (3,3 mL). Purificación: la mezcla orgánica se evaporó hasta sequedad a vacío y a continuación se disolvió en CH2CI2 y lavó con NaOH 1 M, salmuera y agua. La fase orgánica se acidificó con HCI hasta pH = 3 y finalmente
la fase acuosa se basificó con NaOH 1 M hasta la aparición de un precipitado blanco. Rendimiento: 30,5 mg, 16%. Sólido blanco. Pf: 143 - 145 °C. 1H RMN (300 MHz, DMSO- d6) δ 10,29 (s, 1 H) 7,93 (d, J = 4,1 Hz, 1 H), 7,82 (dd, J = 7,9, 1 ,7 Hz, 1 H), 7,78 - 7,75 (m, 2H), 7,66 (d, J = 4,2 Hz, 1 H), 7,40 - 7,35 (m, 2H), 7,31 (ddd, J = 8,6, 7,2, 1 ,7 Hz, 1 H), 7,21 (d, J = 8,5 Hz, 1 H), 7,01 (t, J = 7,4 Hz, 1 H), 4,69 - 4,62 (m, 1 H), 3,02 - 2,93 (m, 2H), 2,62 - 2,53 (m, 2H), 2,01 - 1 ,96 (m, 2H), 1 ,66 - 1 ,57 (m, 2H). 13C RMN (75 MHz, DMSO-d6) 1 δ60,6, 152,9, 143,5, 138,7 (2C), 129,4 (2C), 128,4 (2C), 128,0, 126,9, 125,3, 122,5, 122,0 (2C), 120,8, 114,3, 74,7, 43,6 (2C), 32,2 (2C). HRMS (ESI) m/z: [M + H]+ cale, para C22H22CIN2O2S 413,1085; encontrado 413,1077. Pureza (HPLC): 98,7%. Reagents: N-(4-chlorophenyl)-5-(2-hydroxyphenyl)thiophen-2-carboxamide (15) (0.76 mmol, 250 mg), tert-butyl 4-hydroxypiperidin-1-carboxylate (0.99 mmol, 199 mg), diisopropyl azodicarboxylate (0.99 mmol, 195 pL), triphenylphosphine (0.99 mmol, 259 mg) and 7 mL of THF. 238 mg of tert-butyl 4-(5-((4-chlorophenyl)carbamoyl)thiophen-2-yl)phenoxy)piperidine-1-carboxylate (238 mg) were obtained, which were treated with a CH2CI2 solution (5 mL ) and trifluoroacetic acid (3.3 mL). Purification: The organic mixture was evaporated to dryness in vacuo and then dissolved in CH2Cl2 and washed with 1 M NaOH, brine and water. The organic phase was acidified with HCl until pH = 3 and finally The aqueous phase was basified with 1 M NaOH until a white precipitate appeared. Yield: 30.5 mg, 16%. White solid. mp: 143 - 145 °C. 1 H NMR (300 MHz, DMSO- d6) δ 10.29 (s, 1 H) 7.93 (d, J = 4.1 Hz, 1 H), 7.82 (dd, J = 7.9, 1 .7 Hz, 1 H), 7.78 - 7.75 (m, 2H), 7.66 (d, J = 4.2 Hz, 1 H), 7.40 - 7.35 (m, 2H ), 7.31 (ddd, J = 8.6, 7.2, 1.7 Hz, 1 H), 7.21 (d, J = 8.5 Hz, 1 H), 7.01 (t, J = 7.4 Hz, 1H), 4.69 - 4.62 (m, 1H), 3.02 - 2.93 (m, 2H), 2.62 - 2.53 (m, 2H) , 2.01 - 1 .96 (m, 2H), 1 .66 - 1 .57 (m, 2H). 13 C NMR (75 MHz, DMSO-d6) 1 δ60.6, 152.9, 143.5, 138.7 (2C), 129.4 (2C), 128.4 (2C), 128.0, 126 .9, 125.3, 122.5, 122.0 (2C), 120.8, 114.3, 74.7, 43.6 (2C), 32.2 (2C). HRMS (ESI) m/z: [M + H] + cale, for C22H22CIN2O2S 413.1085; found 413.1077. Purity (HPLC): 98.7%.
N-(4-Clorofenll)-5-(3-(plperldln-4-lloxl)fenll)tlofen-2-carboxamlda (26)N-(4-Chlorofenll)-5-(3-(plperldln-4-lloxl)fenll)tlofen-2-carboxamld (26)
Reactivos: N-(4-clorofenil)-5-(3-hidroxifenil)tiofen-2-carboxamida (16) (1 ,71 mmol, 564 mg), 4-hidroxipiperidin-1-carboxilato de terc-butilo (2,2 mmol, 447 mg), azodicarboxilato de diisopropilo (2,2 mmol, 437 pL), trifenilfosfina (2,2 mmol, 582 mg) y 15 mL de THF. Se obtuvieron 508 mg de 4-(5-((4-clorofenil)carbamoil)tiofen-2-il)fenoxi)piperidina-1- carboxilato de terc-butilo que se trataron con una solución de CH2CI2 (8 mL) y ácido trifluoroacético (2 mL). Purificación: CH2CL2/MeOH 9:1. Rendimiento: 137 mg, 34%. Sólido blanco. Pf: 169 - 171 °C. 1H RMN (300 MHz, DMSO-d6) 10, δ36 (s, 1 H), 8,02 (d, J = 4,0 Hz, 1 H), 7,82 - 7,74 (m, 2H), 7,65 (d, J = 3,9 Hz, 1 H), 7,47 - 7,39 (m, 2H), 7,35 (t, J = 8,1 Hz, 1 H), 7,30 - 7,23 (m, 2H), 7,02 - 6,94 (m, 1 H), 4,64 - 4,37 (m, 1 H), 3,02 - 2,88 (m, 2H), 2,67 - 2,54 (m, 2H), 2,02 - 1 ,86 (m, 2H), 1 ,58 - 1 ,38 (m, 2H). 13C RMN (75 MHz, DMSO-d6) 1 δ59,7, 157,7, 148,5, 138,4, 137,7, 134,3, 130,5, 130,4, 128,6 (2C), 127,4, 124,8, 121 ,8 (2C), 118,1 , 116,1 , 113,0, 73,5, 43,7 (2C), 32,3 (2C). HRMS (ESI) m/z: [M + H]+ cale, para C22H22CIN2O2S 413,1085; encontrado 413,1088. Pureza (HPLC) > 99%. Reagents: N-(4-chlorophenyl)-5-(3-hydroxyphenyl)thiophen-2-carboxamide (16) (1.71 mmol, 564 mg), tert-butyl 4-hydroxypiperidine-1-carboxylate (2.2 mmol, 447 mg), diisopropyl azodicarboxylate (2.2 mmol, 437 pL), triphenylphosphine (2.2 mmol, 582 mg) and 15 mL of THF. 508 mg of tert-butyl 4-(5-((4-chlorophenyl)carbamoyl)thiophen-2-yl)phenoxy)piperidine-1-carboxylate were obtained, which were treated with a solution of CH2CI2 (8 mL) and trifluoroacetic acid. (2 mL). Purification: CH2CL2/MeOH 9:1. Yield: 137 mg, 34%. White solid. mp: 169 - 171 °C. 1 H NMR (300 MHz, DMSO-d6) 10, δ36 (s, 1 H), 8.02 (d, J = 4.0 Hz, 1 H), 7.82 - 7.74 (m, 2H) , 7.65 (d, J = 3.9 Hz, 1 H), 7.47 - 7.39 (m, 2H), 7.35 (t, J = 8.1 Hz, 1 H), 7, 30 - 7.23 (m, 2H), 7.02 - 6.94 (m, 1H), 4.64 - 4.37 (m, 1H), 3.02 - 2.88 (m, 2H) ), 2.67 - 2.54 (m, 2H), 2.02 - 1 .86 (m, 2H), 1 .58 - 1 .38 (m, 2H). 13 C NMR (75 MHz, DMSO-d6) 1 δ59.7, 157.7, 148.5, 138.4, 137.7, 134.3, 130.5, 130.4, 128.6 (2C) , 127.4, 124.8, 121.8 (2C), 118.1, 116.1, 113.0, 73.5, 43.7 (2C), 32.3 (2C). HRMS (ESI) m/z: [M + H] + cale, for C22H22CIN2O2S 413.1085; found 413.1088. Purity (HPLC) > 99%.
N-(4-Clorofenll)-5-(4-(plperldln-4-lloxl)fenll)tlofen-2-carboxamlda (27)N-(4-Chlorofenll)-5-(4-(plperldln-4-lloxl)fenll)tlofen-2-carboxamld (27)
Reactivos: N-(4-clorofenil)-5-(4-hidroxifenil)tiofen-2-carboxamida (17) (3,61 mmol, 1 ,19 g), 4-hidroxipiperidin-1-carboxilato de terc-butilo (3,96 mmol, 798 mg), azodicarboxilato de diisopropilo (4,69 mmol, 925 pL), trifenilfosfina (4,69 mmol, 1 ,23 g) y 20 mL de THF. Se obtuvieron 102 mg de 4-(4-(5-((4-clorofenil)carbamoil)tiofen-2-il)fenoxi)piperidin-1- carboxilato de terc-butilo que se trataron con una solución de CH2CI2 (3,2 mL) y ácido trifluoroacético (0,8 mL). Purificación: CH2CL2/MeOH 9:1. Rendimiento: 45 mg, 55%. Sólido blanco. Pf: 240 - 242 °C. 1H RMN (300 MHz, DMSO-d6) 10, δ40 (s, 1 H), 8,04 (d, J = 4,0 Hz, 1 H), 7,84 - 7,76 (m, 2H), 7,70 - 7,64 (m, 2H), 7,50 (d, J = 4,0 Hz, 1 H), 7,44
- 7,37 (m, 2H), 7,11 - 7,04 (m, 2H), 4,70 - 4,59 (m, 1 H), 3,19 - 3,08 (m, 2H), 2,99 - 2,85 (m, 2H), 2,07 - 1 ,98 (m, 2H), 1 ,81 - 1 ,64 (m, 2H). 13C RMN (75 MHz, DMSO-d6) δ 159,8, 157,2, 148,7, 137,8, 137,4, 130,6, 128,6 (2C), 127,3 (3C), 125,9, 123,3, 121 ,8 (2C), 116,4 (2C), 70,62, 41 ,5 (2C), 28,8 (2C). HRMS (ESI) m/z: [M + H]+ calc. para C22H22CIN2O2S 413,1085; encontrado 413,1080. Pureza (HPLC) > 99%. Reagents: N-(4-chlorophenyl)-5-(4-hydroxyphenyl)thiophen-2-carboxamide (17) (3.61 mmol, 1.19 g), tert-butyl 4-hydroxypiperidine-1-carboxylate (3 .96 mmol, 798 mg), diisopropyl azodicarboxylate (4.69 mmol, 925 pL), triphenylphosphine (4.69 mmol, 1.23 g) and 20 mL of THF. 102 mg of tert-butyl 4-(4-(5-((4-chlorophenyl)carbamoyl)thiophen-2-yl)phenoxy)piperidine-1-carboxylate were obtained and treated with a CH2CI2 solution (3.2 mL) and trifluoroacetic acid (0.8 mL). Purification: CH2CL2/MeOH 9:1. Yield: 45 mg, 55%. White solid. mp: 240 - 242 °C. 1 H NMR (300 MHz, DMSO-d6) 10, δ40 (s, 1 H), 8.04 (d, J = 4.0 Hz, 1 H), 7.84 - 7.76 (m, 2H) , 7.70 - 7.64 (m, 2H), 7.50 (d, J = 4.0 Hz, 1H), 7.44 - 7.37 (m, 2H), 7.11 - 7.04 (m, 2H), 4.70 - 4.59 (m, 1H), 3.19 - 3.08 (m, 2H), 2.99 - 2.85 (m, 2H), 2.07 - 1 .98 (m, 2H), 1 .81 - 1 .64 (m, 2H). 13 C NMR (75 MHz, DMSO-d6) δ 159.8, 157.2, 148.7, 137.8, 137.4, 130.6, 128.6 (2C), 127.3 (3C), 125.9, 123.3, 121.8 (2C), 116.4 (2C), 70.62, 41.5 (2C), 28.8 (2C). HRMS (ESI) m/z: [M + H] + calc. for C 22 H 22 CIN 2 O 2 S 413.1085; found 413.1080. Purity (HPLC) > 99%.
N-Fenetll-5-(2-(plperldln-4-lloxl)fenll)tlofen-2-carboxamlda (28) N-Fenetll-5-(2-(plperldln-4-lloxl)fenll)tlofen-2-carboxamld (28)
Reactivos: N-fenetil-5-(2-hidroxifenil)tiofen-2-carboxamida (18) (0,82 mmol, 265 mg), 4- hidroxipiperidin-1 -carboxilato de terc-butilo (1 ,05 mmol, 211 mg), azodicarboxilato de diisopropilo (1 ,05 mmol, 210 pL), trifenilfosfina (1 ,05 mmol, 275 mg) y 7 mL de THF. Se obtuvieron 237 mg de 4-(5-(fenetilcarbamoil)tiofen-2-il)fenoxi)piperidin-1 -carboxilato de terc-butilo que se trataron con una solución de CH2CI2 (5 mL) y ácido trifluoroacético (3,3 mL). Purificación: CH2Cl2/MeOH 9:1. Rendimiento: 30,5 mg, 16%. Sólido amarillo. Pf: descompone. 1H RMN (300 MHz, DMSO-d6) δ 8,56 (t, J = 5,7 Hz, 1 H), 7,78 (dd, J = 7,9, 1 ,7 Hz, 1 H), 7,67 (d, J = 4,0 Hz, 1 H), 7,59 (d, J = 4,0 Hz, 1 H), 7,33 - 7,18 (m, 7H), 7,02 - 6,97 (m, 1 H), 4,66 - 4,56 (m, 1 H), 3,46 (q, J = 6,9, 6,4 Hz, 2H), 3,02 - 2,94 (m, 2H), 2,84 (t, J = 7,5 Hz, 2H), 2,62 - 2,53 (m, 2H), 2,00 - 1 ,94 (m, 2H), 1 ,65 - 1 ,54 (m, 2H). 13C RMN (75 MHz, DMSO-d6) δ 161 ,5, 152,9, 142,7, 139,5, 139,0, 129,3, 128,7 (2C), 128,4 (2C), 127,9, 127,1 , 126,1 , 125,2, 122,6, 120,8, 114,3, 74,6, 43,7 (2C), 40,8, 35,3, 32,3 (2C). HRMS (ESI) m/z: [M + H]+ cale, para C24H27N2O2S 407,1788; encontrado 407,1782. Pureza (HPLC) > 99%. Reagents: N-phenethyl-5-(2-hydroxyphenyl)thiophen-2-carboxamide (18) (0.82 mmol, 265 mg), tert-butyl 4-hydroxypiperidine-1-carboxylate (1.05 mmol, 211 mg ), diisopropyl azodicarboxylate (1.05 mmol, 210 pL), triphenylphosphine (1.05 mmol, 275 mg) and 7 mL of THF. 237 mg of tert-butyl 4-(5-(phenethylcarbamoyl)thiophen-2-yl)phenoxy)piperidine-1-carboxylate were obtained, which were treated with a solution of CH2CI2 (5 mL) and trifluoroacetic acid (3.3 mL). ). Purification: CH 2 Cl 2 /MeOH 9:1. Yield: 30.5 mg, 16%. Yellow solid. Pf: decompose. 1 H NMR (300 MHz, DMSO-d6) δ 8.56 (t, J = 5.7 Hz, 1 H), 7.78 (dd, J = 7.9, 1.7 Hz, 1 H), 7.67 (d, J = 4.0 Hz, 1 H), 7.59 (d, J = 4.0 Hz, 1 H), 7.33 - 7.18 (m, 7H), 7.02 - 6.97 (m, 1 H), 4.66 - 4.56 (m, 1 H), 3.46 (q, J = 6.9, 6.4 Hz, 2H), 3.02 - 2 .94 (m, 2H), 2.84 (t, J = 7.5 Hz, 2H), 2.62 - 2.53 (m, 2H), 2.00 - 1 .94 (m, 2H), 1 .65 - 1 .54 (m, 2H). 13 C NMR (75 MHz, DMSO-d6) δ 161.5, 152.9, 142.7, 139.5, 139.0, 129.3, 128.7 (2C), 128.4 (2C), 127.9, 127.1, 126.1, 125.2, 122.6, 120.8, 114.3, 74.6, 43.7 (2C), 40.8, 35.3, 32.3 (2 C). HRMS (ESI) m/z: [M + H] + cale, for C24H27N2O2S 407.1788; found 407.1782. Purity (HPLC) > 99%.
N-Fenetll-5-(3-(plperldln-4-lloxl)fenll)tlofen-2-carboxamlda (29) N-Fenetll-5-(3-(plperldln-4-lloxl)fenll)tlofen-2-carboxamld (29)
Reactivos: N-fenetil-5-(3-hidroxifenil)tiofen-2-carboxamida (19) (1 ,39 mmol, 450 mg), 4- hidroxipiperidin-1 -carboxilato de terc-butilo (1 ,81 mmol, 365 mg), azodicarboxilato de diisopropilo (1 ,81 mmol, 357 pL), trifenilfosfina (1 ,81 mmol, 475 mg) y 15 mL de THF. Se obtuvieron 230 mg de 4-(3-(5-(fenetilcarbamoil)tiofen-2-il)fenoxi)piperidin-1- carboxilato de terc-butilo que se trataron con una solución de CH2CI2 (5 mL) y ácido trifluoroacético (2,25 mL). Purificación: no fue necesario ningún proceso de purificación. Rendimiento: 145 mg, 79%. Sólido blanco. Pf: 152 - 154 °C. 1H RMN (300 MHz, DMSO- d6) δ 8,62 (t, J = 5,6 Hz, 1 H), 7,71 (d, J = 4,0 Hz, 1 H), 7,54 (d, J = 3,9 Hz, 1 H), 7,37 - 7,16 (m, 8H), 6,96 (ddd, J = 8,2, 2,4, 1 ,1 Hz, 1 H), 4,52 (tt, J = 8,7, 3,9 Hz, 1 H), 3,46 (dt, J = 8,1 , 6,2 Hz, 2H), 3,05 - 2,92 (m, 2H), 2,84 (t, J = 8,3 Hz, 2H), 2,67 - 2,59 (m, 2H), 2,02 - 1 ,89 (m, 2H), 1 ,58 - 1 ,39 (m, 2H). 13C RMN (75 MHz, DMSO- d6) δ 160,8, 157,6, 147,0, 139,4, 139,0, 134,5, 130,4, 128,8, 128,7 (2C), 128,4 (2C), 126,1 , 124,5, 118,1 ,
115,8, 112,9, 73,1 , 43,3 (20), 40,8, 35,2, 31 ,8 (2C). HRMS (ESI) m/z: [M + H]+ cale, para C24H27N2O2S 407,1788; encontrado 407,1781. Pureza (HPLC): 96,6%. Reagents: N-phenethyl-5-(3-hydroxyphenyl)thiophen-2-carboxamide (19) (1.39 mmol, 450 mg), tert-butyl 4-hydroxypiperidin-1-carboxylate (1.81 mmol, 365 mg ), diisopropyl azodicarboxylate (1.81 mmol, 357 pL), triphenylphosphine (1.81 mmol, 475 mg) and 15 mL of THF. 230 mg of tert-butyl 4-(3-(5-(phenethylcarbamoyl)thiophen-2-yl)phenoxy)piperidine-1-carboxylate were obtained, which were treated with a solution of CH2CI2 (5 mL) and trifluoroacetic acid (2 .25 mL). Purification: no purification process was necessary. Yield: 145 mg, 79%. White solid. mp: 152 - 154 °C. 1 H NMR (300 MHz, DMSO- d 6 ) δ 8.62 (t, J = 5.6 Hz, 1 H), 7.71 (d, J = 4.0 Hz, 1 H), 7.54 (d, J = 3.9 Hz, 1 H), 7.37 - 7.16 (m, 8H), 6.96 (ddd, J = 8.2, 2.4, 1 .1 Hz, 1 H ), 4.52 (tt, J = 8.7, 3.9 Hz, 1H), 3.46 (dt, J = 8.1, 6.2 Hz, 2H), 3.05 - 2.92 (m, 2H), 2.84 (t, J = 8.3 Hz, 2H), 2.67 - 2.59 (m, 2H), 2.02 - 1 .89 (m, 2H), 1 , 58 - 1.39 (m, 2H). 13 C NMR (75 MHz, DMSO- d 6 ) δ 160.8, 157.6, 147.0, 139.4, 139.0, 134.5, 130.4, 128.8, 128.7 (2C ), 128.4 (2C), 126.1, 124.5, 118.1, 115.8, 112.9, 73.1, 43.3 (20), 40.8, 35.2, 31.8 (2C). HRMS (ESI) m/z: [M + H] + cale, for C24H27N2O2S 407.1788; found 407.1781. Purity (HPLC): 96.6%.
N-(4-Morfollnofenll)-5-(2-(plperldln-4-lloxl)fenll)tlofen-2-carboxamlda (30)N-(4-Morfollnofenll)-5-(2-(plperldln-4-lloxl)fenll)tlofen-2-carboxamld (30)
Reactivos: 5-(2-hidroxifenil)-N-(4-morfolinofenil)tiofen-2-carboxamida (20) (2,80 mmol, 786 mg), 4-hidroxipiperidin-1-carboxilato de tere-butilo (3,63 mmol, 729 mg), azodicarboxilato de diisopropilo (3,63 mmol, 714 μL), trifenilfosfina (3,63 mmol, 951 mg) y 15 mL de THF. Se obtuvieron 300 mg de 4-(5-((4-morfolinofenil)carbamoil)tiofen-2- il)fenoxi)piperidin- 1 -carboxilato de tere-butilo que se trataron con una solución de CH2CI2 (10 mL) y ácido trifluoroacético (6,6 mL). Purificación: la mezcla orgánica se acidificó con HCI hasta pH = 3 y a continuación la fase acuosa se basificó con NaOH 1 M hasta la aparición de un precipitado amarillo. El precipitado se extrajo con acetato de etilo. Rendimiento: 57,6 mg, 24%. Sólido amarillo. Pf: 184 - 186 °C. 1H RMN (300 MHz, DMSO-d6) δ 10,05 (s, 1 H), 7,96 (d, J = 4,1 Hz, 1 H), 7,84 (dd, J = 7,9, 1 ,6 Hz, 1 H,), 7,67 (d, J = 4,1 Hz, 1 H), 7,63 - 7,56 (m, 2H), 7,34 (ddd, J = 8,7, 7,1 , 1 ,6 Hz, 1 H), 7,28 - 7,22 (m, 1 H), 7,04 (ddd, J = 8,0, 7,2, 1 ,2 Hz, 1 H), 6,99 - 6,90 (m, 2H), 4,84 - 4,74 (m, 1 H), 3,79 - 3,69 (m, 4H), 3,22 - 3,11 (m, 2H), 3,11 - 3,04 (m, 4H), 2,95 - 2,83 (m, 2H), 2,17 -2,03 (m, 2H), 1 ,90 - 1 ,74 (m, 2H). 13C RMN (75 MHz, DMSO-d6) 159 δ,7, 152,7, 147,5,Reagents: 5-(2-hydroxyphenyl)-N-(4-morpholinophenyl)thiophen-2-carboxamide (20) (2.80 mmol, 786 mg), tere-butyl 4-hydroxypiperidine-1-carboxylate (3.63 mmol, 729 mg), diisopropyl azodicarboxylate (3.63 mmol, 714 μL), triphenylphosphine (3.63 mmol, 951 mg), and 15 mL of THF. 300 mg of tere-butyl 4-(5-((4-morpholinophenyl)carbamoyl)thiophen-2-yl)phenoxy)piperidine-1-carboxylate were obtained, which were treated with a solution of CH2CI2 (10 mL) and trifluoroacetic acid. (6.6 mL). Purification: the organic mixture was acidified with HCl until pH = 3 and then the aqueous phase was basified with 1 M NaOH until the appearance of a yellow precipitate. The precipitate was extracted with ethyl acetate. Yield: 57.6 mg, 24%. Yellow solid. mp: 184 - 186 °C. 1 H NMR (300 MHz, DMSO-d6) δ 10.05 (s, 1 H), 7.96 (d, J = 4.1 Hz, 1 H), 7.84 (dd, J = 7.9 , 1 .6 Hz, 1 H,), 7.67 (d, J = 4.1 Hz, 1 H), 7.63 - 7.56 (m, 2H), 7.34 (ddd, J = 8 .7, 7.1, 1.6 Hz, 1 H), 7.28 - 7.22 (m, 1 H), 7.04 (ddd, J = 8.0, 7.2, 1.2 Hz , 1H), 6.99 - 6.90 (m, 2H), 4.84 - 4.74 (m, 1H), 3.79 - 3.69 (m, 4H), 3.22 - 3 .11 (m, 2H), 3.11 - 3.04 (m, 4H), 2.95 - 2.83 (m, 2H), 2.17 -2.03 (m, 2H), 1 .90 - 1.74 (m, 2H). 13 C NMR (75 MHz, DMSO-d6) 159 δ.7, 152.7, 147.5,
143.1 , 139,4, 131 ,6, 130,9, 129,5, 128,7, 127,9, 125,5, 122,5, 121 ,4 (2C), 115,3 (2C),143.1, 139.4, 131.6, 130.9, 129.5, 128.7, 127.9, 125.5, 122.5, 121.4 (2C), 115.3 (2C),
114.2, 72,3, 66,1 (2C), 48,8 (2C), 42,2 (2C), 29,6 (2C). HRMS (ESI) m/z: [M + H]+ cale, para C26H30N3O3S 464,2002; encontrado 464,2000. Pureza (HPLC) > 99%. 114.2, 72.3, 66.1 (2C), 48.8 (2C), 42.2 (2C), 29.6 (2C). HRMS (ESI) m/z: [M + H] + cale, for C26H30N3O3S 464,2002; found 464,2000. Purity (HPLC) > 99%.
Síntesis de N-fenll-5-(2-(plperldln-4-lloxl)fenll)tlofen-2-carboxamlda (31)Synthesis of N-fenll-5-(2-(plperldln-4-lloxl)fenll)tlofen-2-carboxamld (31)
Sobre una mezcla de (8) 5-bromo-N-feniltiofen-2-carboxamida (1 equiv.), Na2CO3 (2,2 equiv.) y paladio (0) tetrakis(trifenilfosfina) (0,05 equiv.), se añadió una disolución de tolueno, H2O y EtOH (2:1 ,5:1) y 2-(4,4,5,5-tetrametil-1 ,3,2-dioxaborolan-2-il)fenol (1 ,2 equiv.) La mezcla de reacción se burbujeó con argón durante 15 minutos y a continuación se calentó bajo irradiación de microondas (MW) durante 30 minutos a 120ºC . Finalizado este tiempo, el crudo de reacción se adicionó una disolución 1 :1 de AcOEt/H2O y se extrajo con AcOEt. La fase orgánica se lavó con una disolución 1 :1 de disolución saturada de NaCI y H2O, y finalmente se secó sobre Na2SO4 anhidro. A continuación, el desecante se filtró y los volátiles se evaporaron hasta sequedad a vacío obteniéndose un residuo que se utilizó en el siguiente paso. On a mixture of (8) 5-bromo-N-phenylthiophen-2-carboxamide (1 equiv.), Na2CO3 (2.2 equiv.) and palladium (0) tetrakis(triphenylphosphine) (0.05 equiv.), added a solution of toluene, H2O and EtOH (2:1,5:1) and 2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (1,2 equiv .) The reaction mixture was bubbled with argon for 15 minutes and then heated under microwave (MW) irradiation for 30 minutes at 120 ° C. At the end of this time, a 1:1 solution of AcOEt/H 2 O was added to the crude reaction and extracted with AcOEt. The organic phase was washed with a 1:1 solution of saturated NaCl solution and H2O, and finally dried over anhydrous Na2SO4. Next, the desiccant was filtered and the volatiles were evaporated to dryness under vacuum, obtaining a residue that was used in the next step.
A una disolución del residuo anterior (1 equiv.) y trifenilfosfina (1 ,3 equiv.) en THF a 0
ºC se añadió DIAD (1 ,3 equiv.) y 4-hidroxipiperidina-1-carboxilato de tere-butilo (1 ,3 equiv.). Se dejó agitando toda la noche y a continuación se eliminó el disolvente a presión reducida. El residuo sólido blanco obtenido se disolvió en una solución de CH2CI2/TFA 3:2 y se agitó a temperatura ambiente durante 3 horas. Finalizada la desprotección, se evaporó el disolvente a vacío y tras disolver el producto resultante en CH2CI2, se basificó con NaHCO3 para obtener el producto en forma neutra. A continuación, los volátiles se evaporaron hasta sequedad a vacío y el residuo resultante se purificó por cromatografía en columna utilizando como eluyente una mezcla CH2Cl2/MeOH 9:1. Rendimiento: 90,5 mg, 35%. Sólido blanco. Pf: 122 - 124 ºC . 1H RMN (300 MHz, DMSO-d6) 1 δ0,28 (s, 1 H), 8,07 (d, J = 4,1 Hz, 1 H), 7,87 (dd, J = 7,9, 1 ,6 Hz, 1 H), 7,81 - 7,73 (m, 2H), 7,70 (d, J = 4,1 Hz, 1 H), 7,41 - 7,31 (m, 3H), 7,29 (d, J = 7,7 Hz, 1 H), 7,12 (d, J = 7,5 Hz, 1 H), 7,07 (dd, J = 7,8, 1 ,2 Hz, 1 H), 4,91 (dt, J = 7,9, 4,0 Hz, 1 H), 3,30 (m, 2H), 3,13 (m, 2H), 2,21 (m, 2H), 1 ,98 (m, 2H). 13C RMN (75 MHz, DMSO- d6) δ 160,5, 152,8, 143,7, 139,5, 139,2, 130,1 , 129,0, 128,9, 128,7, 126,1 , 124,0, 121 ,8, 120,6, 114,5, 70,7, 41 ,3, 27,7. HRMS (ESI) m/z: [M + H]+ cale, para C22H23N2O2S 379,1475; encontrado 379,1473. Pureza (HPLC) > 99%. To a solution of the previous residue (1 equiv.) and triphenylphosphine (1.3 equiv.) in THF at 0 ºC, DIAD (1.3 equiv.) and tere-butyl 4-hydroxypiperidine-1-carboxylate (1.3 equiv.) were added. It was left stirring overnight and then the solvent was removed under reduced pressure. The white solid residue obtained was dissolved in a 3:2 CH2CI2/TFA solution and stirred at room temperature for 3 hours. Once the deprotection was completed, the solvent was evaporated under vacuum and after dissolving the resulting product in CH2CI2, it was basified with NaHCO 3 to obtain the product in neutral form. The volatiles were then evaporated to dryness in vacuo and the resulting residue was purified by column chromatography using a 9:1 CH 2 Cl 2 /MeOH mixture as eluent. Yield: 90.5 mg, 35%. White solid. Pf: 122 - 124 º C. 1 H NMR (300 MHz, DMSO-d6) 1 δ0.28 (s, 1 H), 8.07 (d, J = 4.1 Hz, 1 H), 7.87 (dd, J = 7.9 , 1 .6 Hz, 1 H), 7.81 - 7.73 (m, 2H), 7.70 (d, J = 4.1 Hz, 1 H), 7.41 - 7.31 (m, 3H), 7.29 (d, J = 7.7 Hz, 1 H), 7.12 (d, J = 7.5 Hz, 1 H), 7.07 (dd, J = 7.8, 1 .2 Hz, 1 H), 4.91 (dt, J = 7.9, 4.0 Hz, 1 H), 3.30 (m, 2H), 3.13 (m, 2H), 2.21 (m, 2H), 1 .98 (m, 2H). 13 C NMR (75 MHz, DMSO- d6) δ 160.5, 152.8, 143.7, 139.5, 139.2, 130.1, 129.0, 128.9, 128.7, 126, 1, 124.0, 121.8, 120.6, 114.5, 70.7, 41.3, 27.7. HRMS (ESI) m/z: [M + H] + cale, for C22H23N2O2S 379.1475; found 379.1473. Purity (HPLC) > 99%.
Síntesis de N-fenll-5-(3-(plperldln-4-lloxl)fenll)tlofen-2-carboxamlda (32)Synthesis of N-fenll-5-(3-(plperldln-4-lloxl)fenll)tlofen-2-carboxamld (32)
Sobre una mezcla de (8) 5-bromo-N-feniltiofen-2-carboxamida (1 equiv.), Na2CO3 (2,2 equiv.) y paladio (0) tetrakis(trifenilfosfina) (0,05 equiv.), se añadió una disolución de tolueno, H2O y EtOH (2:1 ,5:1) y 3-(4,4,5,5-tetrametil-1 ,3,2-dioxaborolan-2-il)fenol (1 ,2 equiv.) La mezcla de reacción se burbujeó con argón durante 15 minutos y a continuación se calentó bajo irradiación de microondas (MW) durante 30 minutos a 120 ºC. Finalizado este tiempo, el crudo de reacción se adicionó una disolución 1 :1 de AcOEt/H2O y se extrajo con AcOEt. La fase orgánica se lavó con una disolución 1 :1 de disolución saturada de NaCI y H2O, y finalmente se secó sobre Na2SO4 anhidro. A continuación, el desecante se filtró y los volátiles se evaporaron hasta sequedad a vacío obteniéndose un residuo que se utilizó en el siguiente paso. On a mixture of (8) 5-bromo-N-phenylthiophen-2-carboxamide (1 equiv.), Na 2 CO 3 (2.2 equiv.) and palladium (0) tetrakis(triphenylphosphine) (0.05 equiv. ), a solution of toluene, H 2 O and EtOH (2:1,5:1) and 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol was added (1.2 equiv.) The reaction mixture was bubbled with argon for 15 minutes and then heated under microwave irradiation (MW) for 30 minutes at 120 ºC . After this time, a solution 1 was added to the reaction crude. :1 AcOEt/H 2 O and extracted with AcOEt. The organic phase was washed with a 1:1 solution of saturated NaCl solution and H 2 O, and finally dried over anhydrous Na 2 SO 4 . Next, the desiccant was filtered and the volatiles were evaporated to dryness under vacuum, obtaining a residue that was used in the next step.
A una disolución del residuo anterior (1 equiv.) y trifenilfosfina (1 ,3 equiv.) en THF a 0 ºC se añadió DIAD (1 ,3 equiv.) y 4-hidroxipiperidina-1-carboxilato de tere-butilo (1 ,3 equiv.). Se dejó agitando toda la noche y a continuación se eliminó el disolvente a presión reducida. El residuo sólido blanco obtenido se disolvió en una solución de CH2CI2/TFA 3:2 y se agitó a temperatura ambiente durante 3 horas. Finalizada la desprotección, se evaporó el disolvente a vacío y tras disolver el producto resultante en
CH2CI2 se basificó con NaHCO3 para obtener el producto en forma neutra. A continuación, los volátiles se evaporaron hasta sequedad a vacío y el residuo resultante se purificó por cromatografía en columna utilizando como eluyente una mezcla CH2Cl2/MeOH 9:1. Rendimiento: 80,8 mg, 32%. Sólido blanco. Pf: 151 - 153 °C. 1H RMN (300 MHz, DMSO-d6) 1 δ0,27 (s, 1 H), 8,05 (d, J = 4,0 Hz, 1 H), 7,81 - 7,70 (m, 2H), 7,65 (d, J = 4,0 Hz, 1 H), 7,36 (m, 3H), 7,28 (m, 2H), 7,21 - 7,05 (m, 1 H), 6,99 (d, J = 8,1 Hz, 1 H), 4,56 (dt, J = 8,8, 4,7 Hz, 1 H), 3,08 - 2,90 (m, 2H), 2,67 (m, 2H), 1 ,96 (dd, J = 12,9, 4,0 Hz, 2H), 1 ,53 (qd, J = 9,2, 4,7 Hz, 2H). 13C RMN (75 MHz, DMSO-d6) 160, δ0, 157,9, 148,5, 139,2, 139,0, 134,8, 130,9, 130,5, 129,0, 125,1 , 124,1 , 120,7, 118,6, 116,4, 113,4, 73,2, 43,5, 31 ,9. HRMS (ESI) m/z: [M + H]+ cale, para C22H23N2O2S 379,1475; encontrado 379,1475. Pureza (HPLC) > 99%. To a solution of the previous residue (1 equiv.) and triphenylphosphine (1.3 equiv.) in THF at 0 ºC was added DIAD (1.3 equiv.) and tere-butyl 4-hydroxypiperidine-1-carboxylate (1 .3 equiv.). It was left stirring overnight and then the solvent was removed under reduced pressure. The white solid residue obtained was dissolved in a 3:2 CH2CI2/TFA solution and stirred at room temperature for 3 hours. Once the deprotection was completed, the solvent was evaporated in vacuo and after dissolving the resulting product in CH2CI2 was basified with NaHCO3 to obtain the product in neutral form. The volatiles were then evaporated to dryness in vacuo and the resulting residue was purified by column chromatography using a 9:1 CH 2 Cl2/MeOH mixture as eluent. Yield: 80.8 mg, 32%. White solid. mp: 151 - 153 °C. 1 H NMR (300 MHz, DMSO-d6) 1 δ0.27 (s, 1 H), 8.05 (d, J = 4.0 Hz, 1 H), 7.81 - 7.70 (m, 2H ), 7.65 (d, J = 4.0 Hz, 1H), 7.36 (m, 3H), 7.28 (m, 2H), 7.21 - 7.05 (m, 1H) , 6.99 (d, J = 8.1 Hz, 1 H), 4.56 (dt, J = 8.8, 4.7 Hz, 1 H), 3.08 - 2.90 (m, 2H ), 2.67 (m, 2H), 1 .96 (dd, J = 12.9, 4.0 Hz, 2H), 1 .53 (qd, J = 9.2, 4.7 Hz, 2H) . 13 C NMR (75 MHz, DMSO-d6) 160, δ0, 157.9, 148.5, 139.2, 139.0, 134.8, 130.9, 130.5, 129.0, 125.1 , 124.1, 120.7, 118.6, 116.4, 113.4, 73.2, 43.5, 31.9. HRMS (ESI) m/z: [M + H] + cale, for C22H23N2O2S 379.1475; found 379.1475. Purity (HPLC) > 99%.
Síntesis de N-fenll-5-(4-(plperldln-4-lloxl)fenll)tlofen-2-carboxamlda (33)Synthesis of N-fenll-5-(4-(plperldln-4-lloxl)fenll)tlofen-2-carboxamld (33)
Sobre una mezcla de (8) 5-bromo-N-feniltiofen-2-carboxamida (1 equiv.), Na2CO3 (2,2 equiv.) y paladio (0) tetrakis(trifenilfosfina) (0,05 equiv.), se añadió una disolución de tolueno, H2O y EtOH (2:1 ,5:1) y 4-(4,4,5,5-tetrametil-1 ,3,2-dioxaborolan-2-il)fenol (1 ,2 equiv.) La mezcla de reacción se burbujeó con argón durante 15 minutos y a continuación se calentó bajo irradiación de microondas (MW) durante 30 minutos a 120ºC . Finalizado este tiempo, el crudo de reacción se adicionó una disolución 1 :1 de AcOEt/H2O y se extrajo con AcOEt. La fase orgánica se lavó con una disolución 1 :1 de disolución saturada de NaCI y H2O, y finalmente se secó sobre Na2SO4 anhidro. A continuación, el desecante se filtró y los volátiles se evaporaron hasta sequedad a vacío obteniéndose un residuo que se utilizó en el siguiente paso. On a mixture of (8) 5-bromo-N-phenylthiophen-2-carboxamide (1 equiv.), Na2CO3 (2.2 equiv.) and palladium (0) tetrakis(triphenylphosphine) (0.05 equiv.), added a solution of toluene, H2O and EtOH (2:1,5:1) and 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (1,2 equiv .) The reaction mixture was bubbled with argon for 15 minutes and then heated under microwave (MW) irradiation for 30 minutes at 120 ° C. At the end of this time, a 1:1 solution of AcOEt/H 2 O was added to the crude reaction and extracted with AcOEt. The organic phase was washed with a 1:1 solution of saturated NaCl solution and H2O, and finally dried over anhydrous Na2SO 4 . Next, the desiccant was filtered and the volatiles were evaporated to dryness under vacuum, obtaining a residue that was used in the next step.
A una disolución del residuo anterior (1 equiv.) y trifenilfosfina (1 ,3 equiv.) en THF a 0ºC se añadió DIAD (1 ,3 equiv.) y 4-hidroxipiperidina-1-carboxilato de tere-butilo (1 ,3 equiv.). Se dejó agitando toda la noche y a continuación se eliminó el disolvente a presión reducida. El residuo sólido blanco obtenido se disolvió en una solución de CH2CI2/TFA 3:2 y se agitó a temperatura ambiente durante 3 horas. Finalizada la desprotección, se evaporó el disolvente a vacío y tras disolver el producto resultante en CH2CI2 se basificó con NaHCOa para obtener el producto en forma neutra. A continuación, los volátiles se evaporaron hasta sequedad a vacío y el residuo resultante se purificó por cromatografía en columna utilizando como eluyente una mezcla CH2Cl2/MeOH 9:1. Rendimiento: 96,3 mg, 38%. Sólido blanco. Pf: 192 - 194 ºC. 1H RMN (300 MHz, DMSO-d6) 1 δ0,22 (s, 1 H), 8,01 (d, J = 4,0 Hz, 1 H), 7,79 - 7,68 (m, 2H), 7,68 - 7,57 (m, 2H), 7,49 (d, J = 3,9 Hz, 1 H), 7,36 (t, J = 7,9 Hz, 2H), 7,15 - 7,07 (m, 1 H),
7,07 - 6,94 (m, 2H), 4,69 - 4,38 (m, 1 H), 2,98 (dd, J = 12,6, 4,8 Hz, 2H), 2,65 (ddd, J = 12,8, 9,9, 2,9 Hz, 2H), 2,04 - 1 ,86 (m, 2H), 1 ,51 (m, 2H). 13C RMN (75 MHz, DMSO d6) 160,1 , 157,9, 148,9, 139,1 , 137,9, 130,7, 129,0, 127,6, 125,9, 124,0, 123,5, 120,7, 116,7, 73,3, 43,6, 31 ,9. HRMS (ESI) m/z: [M + H]+ calc, para C22H23N2O2S 379,1475; encontrado 379,1469. Pureza (HPLC) > 99%. To a solution of the previous residue (1 equiv.) and triphenylphosphine (1.3 equiv.) in THF at 0 ºC was added DIAD (1.3 equiv.) and tere-butyl 4-hydroxypiperidine-1-carboxylate (1 .3 equiv.). It was left stirring overnight and then the solvent was removed under reduced pressure. The white solid residue obtained was dissolved in a 3:2 CH2CI2/TFA solution and stirred at room temperature for 3 hours. Once the deprotection was completed, the solvent was evaporated in vacuo and after dissolving the resulting product in CH2CI2, it was basified with NaHCOa to obtain the product in neutral form. The volatiles were then evaporated to dryness in vacuo and the resulting residue was purified by column chromatography using a 9:1 CH 2 Cl 2 /MeOH mixture as eluent. Yield: 96.3 mg, 38%. White solid. mp: 192 - 194 º C. 1 H NMR (300 MHz, DMSO-d6) 1 δ0.22 (s, 1 H), 8.01 (d, J = 4.0 Hz, 1 H), 7.79 - 7.68 (m, 2H), 7.68 - 7.57 (m, 2H), 7.49 (d, J = 3.9 Hz, 1 H), 7.36 (t, J = 7, 9Hz, 2H), 7.15 - 7.07 (m, 1H), 7.07 - 6.94 (m, 2H), 4.69 - 4.38 (m, 1H), 2.98 (dd, J = 12.6, 4.8 Hz, 2H), 2.65 (ddd, J = 12.8, 9.9, 2.9 Hz, 2H), 2.04 - 1 .86 (m, 2H), 1 .51 (m, 2H). 13 C NMR (75 MHz, DMSO d6) 160.1, 157.9, 148.9, 139.1, 137.9, 130.7, 129.0, 127.6, 125.9, 124.0, 123.5, 120.7, 116.7, 73.3, 43.6, 31.9. HRMS (ESI) m/z: [M + H] + calc, for C22H23N2O2S 379.1475; found 379.1469. Purity (HPLC) > 99%.
Síntesis de N-(2-(1 -bencllplperldln-4-ll)etll)-5-(3-metoxlfenll)tlofen-2-carboxamlda (34) Synthesis of N-(2-(1 -bencllplperldln-4-ll)etll)-5-(3-methoxlfenll)tlofen-2-carboxamlda (34)
Sobre una disolución de trifenilfosfina (2,07 mmol, 542 mg) en THF a 0 °C se añadió DIAD (2,07 mmol, 408 pL). La mezcla de reacción se agitó a temperatura ambiente hasta que apareció turbidez blanca. Tras enfriar de nuevo a 0 ºC , se añadió una solución de N-(2-(1-bencilpiperidin-4-il)etil)-5-(3-hidroxifenil)tiofen-2-carboxamida (10) (1 ,59 mmol, 669 mg), 4-hidroxipiperidin-1-carboxilato de tere-butilo (2,07 mmol, 415 mg y se agitó durante toda la noche. A continuación, se eliminó el disolvente a presión reducida y el residuo resultante se purificó por cromatografía en columna utilizando como eluyente una mezcla CH2CL2/MeOH 9:1. Rendimiento: 80,1 mg, 12 %. Sólido blanco. Pf: 99 - 101ºC . 1H RMN (300 MHz, DMSO-d6) 8 δ,46 (t, J = 5,6 Hz, 1 H), 7,72 (d, J = 4,0 Hz, 1 H), 7,54 (d, J = 3,9 Hz, 1 H), 7,39 - 7,18 (m, 8H), 6,94 (ddd, J = 8,2, 2,5, 1 ,0 Hz, 1 H), 3,81 (s, 3H), 3,43 (s, 2H), 3,26 (q, J = 6,7 Hz, 2H), 2,86 - 2,69 (m, 2H), 1 ,98 - 1 ,81 (m, 2H), 1 ,76 - 1 ,59 (m, 2H), 1 ,46 (q, J = 6,9 Hz, 2H), 1 ,36 - 1 ,24 (m, 1 H), 1 ,23 - 1 ,04 (m, 2H). 13C RMN (75 MHz, DMSO-d6) 160,7, 159,8, 146,9, 139,2, 134,5, 130,4, 128,8, 128,7, 128,1 (4C), 126,8, 124,5, 118,0, 114,1 , 110,9, 62,5, 55,2, 53,2 (2C), 36,8, 35,9, 32,9 (2C), 31 ,8. HRMS (ESI) m/z: [M + H]+ cale, para C26H31N2O2S 435,2101 ; encontrado 435,2099. Pureza (HPLC): 97,7 %. DIAD (2.07 mmol, 408 pL) was added to a solution of triphenylphosphine (2.07 mmol, 542 mg) in THF at 0 °C. The reaction mixture was stirred at room temperature until white turbidity appeared. After cooling again to 0 ° C, a solution of N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-(3-hydroxyphenyl)thiophen-2-carboxamide (10) was added (1.59 mmol, 669 mg), tere-butyl 4-hydroxypiperidine-1-carboxylate (2.07 mmol, 415 mg and stirred overnight. The solvent was then removed under reduced pressure and the resulting residue was purified by Column chromatography using a CH2CL2/MeOH 9:1 mixture as eluent. Yield: 80.1 mg, 12% White solid. mp: 99 - 101 º C. 46 (t, J = 5.6 Hz, 1 H), 7.72 (d, J = 4.0 Hz, 1 H), 7.54 (d, J = 3.9 Hz, 1 H), 7 .39 - 7.18 (m, 8H), 6.94 (ddd, J = 8.2, 2.5, 1 .0 Hz, 1 H), 3.81 (s, 3H), 3.43 ( s, 2H), 3.26 (q, J = 6.7 Hz, 2H), 2.86 - 2.69 (m, 2H), 1 .98 - 1 .81 (m, 2H), 1 .76 - 1 .59 (m, 2H), 1 .46 (q, J = 6.9 Hz, 2H), 1 .36 - 1 .24 (m, 1 H), 1 .23 - 1 .04 (m, 2H ). (4C), 126.8, 124.5, 118.0, 114.1, 110.9, 62.5, 55.2, 53.2 (2C), 36.8, 35.9, 32.9 (2C), 31 .8. HRMS (ESI) m/z: [M + H] + cale, for C26H31N2O2S 435.2101 ; found 435.2099. Purity (HPLC): 97.7%.
Síntesis de N-(2-(1 -bencllplperldln-4-ll)etll)-5-(4-metoxlfenll)tlofen-2-carboxamlda (35) Synthesis of N-(2-(1 -bencllplperldln-4-ll)etll)-5-(4-methoxlfenll)tlofen-2-carboxamlda (35)
Sobre una solución de trifenilfosfina (1 ,15 mmol, 334 mg) en THF a 0 °C se añadió DIAD (1 ,15 mmol, 266 pL). La mezcla de reacción se agitó a temperatura ambiente hasta que apareció turbidez blanca. Tras enfriar de nuevo a 0ºC , se añadió una solución de N-(2- (1-bencilpiperidin-4-il)etil)-5-(4-hidroxifenil)tiofen-2-carboxamida (11) (1 ,04 mmol, 438 mg) y 4-hidroxipiperidina-1-carboxilato de tere-butilo (1 ,15 mmol, 230 mg) en THF y se agitó durante toda la noche. A continuación, se eliminó el disolvente a presión reducida y el residuo resultante se purificó por cromatografía en columna utilizando como eluyente una mezclaCH2CI2 /MeOH 9:1. Rendimiento: 70,3 mg, 16 %. Sólido beige. Pf:
163 - 165 °C. 1H RMN (300 MHz, DMSO-d6) 8, δ41 (t, J = 5,6 Hz, 1 H), 7,68 (d, J = 3,9 Hz, 1 H), 7,66 - 7,58 (m, 2H), 7,38 (d, J = 3,9 Hz, 1 H), 7,36 - 7,20 (m, 5H), 7,04 - 6,95 (m, 2H), 3,79 (s, 3H), 3,44 (s, 2H), 3,26 (q, J = 6,7 Hz, 2H), 2,86 - 2,71 (m, 2H), 2,00 - 1 ,82 (m, 2H), 1 ,74 - 1 ,58 (m, 2H), 1 ,53 - 1 ,38 (m, 2H), 1 ,37 - 1 ,24 (m, 1 H), 1 ,25 - 1 ,09 (m, 2H). 13C RMN (75 MHz, DMSO-d6) 16 δ0,9, 159,5, 147,3, 138,0, 128,8, 128,1 (5C), 127,0 (2C), 126,8, 125,9, 122,9, 114,6 (2C), 62,4, 55,3, 53,2 (2C), 36,7, 35,9, 32,9, 31 ,8 (2C). HRMS (ESI) m/z: [M +H]+calc. para C26H31N2O2S 435,2101 ; encontrado 435,2102. Pureza (HPLC): 96,4 %. DIAD (1.15 mmol, 266 pL) was added to a solution of triphenylphosphine (1.15 mmol, 334 mg) in THF at 0 °C. The reaction mixture was stirred at room temperature until white turbidity appeared. After cooling again to 0 ° C, a solution of N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-(4-hydroxyphenyl)thiophen-2-carboxamide (11) was added (1.04 mmol, 438 mg) and tere-butyl 4-hydroxypiperidine-1-carboxylate (1.15 mmol, 230 mg) in THF and stirred overnight. The solvent was then removed under reduced pressure and the resulting residue was purified by column chromatography using a 9:1 CH2CI2/MeOH mixture as eluent. Yield: 70.3 mg, 16%. Solid beige. Pf: 163 - 165°C. 1 H NMR (300 MHz, DMSO-d6) 8, δ41 (t, J = 5.6 Hz, 1 H), 7.68 (d, J = 3.9 Hz, 1 H), 7.66 - 7 .58 (m, 2H), 7.38 (d, J = 3.9 Hz, 1 H), 7.36 - 7.20 (m, 5H), 7.04 - 6.95 (m, 2H) , 3.79 (s, 3H), 3.44 (s, 2H), 3.26 (q, J = 6.7 Hz, 2H), 2.86 - 2.71 (m, 2H), 2, 00 - 1.82 (m, 2H), 1.74 - 1.58 (m, 2H), 1.53 - 1.38 (m, 2H), 1.37 - 1.24 (m, 1H) , 1 .25 - 1 .09 (m, 2H). 13 C NMR (75 MHz, DMSO-d6) 16 δ0.9, 159.5, 147.3, 138.0, 128.8, 128.1 (5C), 127.0 (2C), 126.8, 125.9, 122.9, 114.6 (2C), 62.4, 55.3, 53.2 (2C), 36.7, 35.9, 32.9, 31.8 (2C). HRMS (ESI) m/z: [M +H]+calc. for C26H31N2O2S 435.2101 ; found 435.2102. Purity (HPLC): 96.4%.
Ejemplo 2: Estudio de la actividad antiviral de los compuestos de fórmula (I) frente al virus del Ébola ÍEBOV) Example 2: Study of the antiviral activity of the compounds of formula (I) against the Ebola virus (IEBOV)
Materiales y Métodos Materials and methods
Líneas celulares cell lines
Las células de riñón embrionario humano 293T/17 (ATCC-CRL-11268), las células de adenocarcinoma cervical humano HeLa (ATCC-CCL-2), de riñón de hámster bebé (BHK-21/WI-2, Kerafast # EH1011) y las células de riñón del mono verde africano Cercopithecus aethiops (VeroE6) se cultivaron en medio DMEM suplementado con 10% de FBS, 1% L -glutamina y 25 pg/mL de gentamicina en incubador a 37 ‘C en presencia de un 5% de CO2. Human Embryonic Kidney 293T/17 Cells (ATCC-CRL-11268), Human Cervical Adenocarcinoma HeLa Cells (ATCC-CCL-2), Baby Hamster Kidney (BHK-21/WI-2, Kerafast #EH1011) and kidney cells of the African green monkey Cercopithecus aethiops (VeroE6) were cultured in DMEM medium supplemented with 10% FBS, 1% L -glutamine and 25 pg/mL gentamicin in incubator at 37 'C in the presence of 5% of CO2.
Producción de virus recombinantes (HIV-May-luc) basados en el virus de inmunodeflciencla humana (VIH), con envolturas (GP/G) de virus Ebola (ZEBOV) Mayings o Virus de la Estomatitis Vesicular (VSV) y que expresan luclferasa tras la infección de células susceptibles. Production of recombinant viruses (HIV-May-luc) based on the human immunodeficiency virus (HIV), with envelopes (GP/G) of Ebola virus (ZEBOV), Mayings or Vesicular Stomatitis Virus (VSV) and that express lucferase after infection of susceptible cells.
Se generaron partículas virales basadas en el virus VIH, pseudotipadas con la glicoproteína de la cepa Ebola-Mayinga (GeneBank: U23187.1) (ZEBOV-GP) o bien la glicoproteína del Virus de la Estomatitis Vesicular (VSV-G) (GenBank: X03633.1) que expresan la proteína luciferasa tras la infección de células susceptibles. Viral particles based on the HIV virus were generated, pseudotyped with the glycoprotein of the Ebola-Mayinga strain (GeneBank: U23187.1) (ZEBOV-GP) or the glycoprotein of the Vesicular Stomatitis Virus (VSV-G) (GenBank: X03633.1) that express the luciferase protein after infection of susceptible cells.
La producción de estas partículas virales se llevó a cabo mediante el método de transfección con cloruro de calcio (Life Technologies, Carlsbad, CA, EE. UU.) en células 293T añadiendo los plásmidos: pNL4.3.Luc.RE- (Programa de reactivos del SIDA de NIH, División de SIDA del Dr. Nathaniel Landau) y la respectiva glicoproteína de cada virus.
Los sobrenadantes que contenían los virus recombinantes se recogieron 48 h después de la transfección, se centrifugaron a 1200 rpm durante 10 minutos a temperatura ambiente para eliminar los restos celulares y se almacenaron congelados a -80ºC en alícuotas. The production of these viral particles was carried out by the calcium chloride transfection method (Life Technologies, Carlsbad, CA, USA) in 293T cells by adding the plasmids: pNL4.3.Luc.RE- (Program of NIH AIDS reagents, Dr. Nathaniel Landau's Division of AIDS) and the respective glycoprotein of each virus. Supernatants containing the recombinant viruses were collected 48 h after transfection, centrifuged at 1200 rpm for 10 min at room temperature to remove cellular debris, and stored frozen at −80 ° C in aliquots.
Los títulos infecciosos de cada pseudotipo viral se estimaron realizando diluciones seriadas (diluciones en serie 1 :5 por triplicado) de los sobrenadantes virales e infectando células 293T. La actividad luciferasa se determinó 48 h después de la infección mediante el ensayo de luciferasa (Luciferase Assay System, Promega, Madison, Wl) utilizando para la lectura el luminómetro GloMax® Navigator Microplate Luminometer (Promega). Infectious titers of each viral pseudotype were estimated by performing serial dilutions (1:5 serial dilutions in triplicate) of the viral supernatants and infecting 293T cells. Luciferase activity was determined 48 h after infection by the luciferase assay (Luciferase Assay System, Promega, Madison, Wl) using the GloMax® Navigator Microplate Luminometer (Promega) for reading.
Producción de virus recomblnantes (VSV-Eb-luc) basados en el virus de la Estomatitis Vesicular (VSV), con envolturas (GP/G) de virus Ebola (ZEBOV) Maylnga o Virus de la Estomatitis Vesicular (VSV) y que expresan luciferasa tras la Infección de células susceptibles. Production of recombinant viruses (VSV-Eb-luc) based on the Vesicular Stomatitis Virus (VSV), with envelopes (GP/G) of Ebola virus (ZEBOV) Maylnga or Vesicular Stomatitis Virus (VSV) and expressing luciferase after infection of susceptible cells.
Para la generación de pseudotipos basados en el virus de la Estomatitis Vesicular, se transfectaron células BHK-21 (Kerafast) con plásmidos que codifican la proteína de la envoltura del virus Ebola (cepa Mayinga) o la propia glicoproteína del Virus de la Estomatitis Vesicular (G) empleando Lipofectamina 3000 (Thermo Fisher Scientific). Al cabo de 24 h se inoculan las células con pseudotipos recombinantes de VSV-delta G- luc, deficientes en su capacidad replicativa (MOI: 3-5) que contiene el gen de la luciferasa de luciérnaga en sustitución de la propia glicoproteína del virus, permitiendo así la incorporación de glicoproteínas heterólogas en las nuevas partículas virales que se generan. For the generation of pseudotypes based on the Vesicular Stomatitis virus, BHK-21 cells (Kerafast) were transfected with plasmids that encode the envelope protein of the Ebola virus (Mayinga strain) or the Vesicular Stomatitis Virus glycoprotein itself ( G) using Lipofectamine 3000 (Thermo Fisher Scientific). After 24 h, the cells are inoculated with recombinant pseudotypes of VSV-delta G-luc, deficient in their replicative capacity (MOI: 3-5) that contain the firefly luciferase gene in replacement of the virus's own glycoprotein. thus allowing the incorporation of heterologous glycoproteins into the new viral particles that are generated.
Tras 1 h de incubación a 37 ºC, se retira el inóculo, se lavan varias veces las células con PBS y finalmente se añade medio fresco. After 1 h of incubation at 37ºC , the inoculum is removed, the cells are washed several times with PBS and finally fresh medium is added.
Las partículas virales se recogen tras 20-24 h post- inoculación, se retiran los restos celulares mediante centrifugación (1200 rpm, 10 minutos) y se guardan alicuotados en congelador de -80 ºC. The viral particles are collected after 20-24 h post-inoculation, the cell debris is removed by centrifugation (1200 rpm, 10 minutes) and stored in aliquots in a -80 º C freezer.
El cálculo del título infeccioso (dosis infecciosa por mililitro), se realizó mediante dilución límite del stock de virus utilizando células Vero E6 como línea susceptible. La actividad luciferasa de determinó 24 h tras la infección mediante ensayo de luciferasa (Steady-
Gio® Luciferase Assay System, Promega) utilizando para la lectura el luminómetro GloMax® Navigator Microplate Luminometer (Promega). The calculation of the infectious titer (infectious dose per milliliter) was carried out by limiting dilution of the virus stock using Vero E6 cells as a susceptible line. Luciferase activity was determined 24 h after infection by luciferase assay (Steady- Gio® Luciferase Assay System, Promega) using the GloMax® Navigator Microplate Luminometer (Promega) for reading.
Cribado de compuestos inhibidores de la infección por virus Ébola Screening of compounds that inhibit Ebola virus infection
El cribado de compuestos inhibidores de la entrada del virus Ébola, se realizó infectando células 293T con pseudotipos ZEBOV-GP en placas de 96 pocilios en presencia de cada compuesto a una concentración final de 10 pM. Screening for compounds that inhibit Ebola virus entry was performed by infecting 293T cells with ZEBOV-GP pseudotypes in 96-well plates in the presence of each compound at a final concentration of 10 pM.
Las células 293T (2x104 células/pocillo) se incubaron a 37 ºC durante 1 h con cada uno de los compuestos y luego se expusieron a 5000 TCID (dosis infecciosa de cultivo) de los virus recombinantes mencionados anteriormente. Después de 48 h de incubación, las células se lavaron con PBS, se lisaron mediante la adición de Steady-Glo Lyssis Buffer (Promega) y se midió la luz en un sistema de detección GloMax®-Multi + (Promega, Madison, Wl, EE. UU.) con el sistema de ensayo de luficerasa (Promega, Madison, Wl). 293T cells (2x104 cells/well) were incubated at 37 ° C for 1 h with each of the compounds and then exposed to 5000 TCID (culture infectious dose) of the above-mentioned recombinant viruses. After 48 h of incubation, cells were washed with PBS, lysed by the addition of Steady-Glo Lyssis Buffer (Promega), and light was measured on a GloMax®-Multi+ detection system (Promega, Madison, WI, USA) with the luficerase assay system (Promega, Madison, Wl).
Como control de inhibición específica, se utilizó el virus recombinante VSV-G en las mismas condiciones y en paralelo a la infección con pseudotipos de Ébola. As a specific inhibition control, the recombinant VSV-G virus was used under the same conditions and in parallel to the infection with Ebola pseudotypes.
Los valores de inhibición se calcularon a partir de 3 experimentos independientes (n=6) y se plasman como la media aritmética. Inhibition values were calculated from 3 independent experiments (n=6) and are expressed as the arithmetic mean.
Los compuestos cuya actividad inhibitoria frente a pseudotipos ZEBOV-GP redujeron la infección más de un 80% a una concentración final de 10 pM, fueron seleccionados para posteriores análisis (CI50 y citotoxicidad). Compounds whose inhibitory activity against ZEBOV-GP pseudotypes reduced infection by more than 80% at a final concentration of 10 pM were selected for subsequent analyzes (IC50 and cytotoxicity).
Cálculo de los valores de CI50 Calculation of IC50 values
Para calcular el valor CI50, se emplearon concentraciones seriadas de cada compuesto desde 10 pM a 10 nM. To calculate the IC50 value, serial concentrations of each compound from 10 pM to 10 nM were used.
Los valores de inhibición se calcularon utilizando la media de 3 experimentos independientes (n=6) empleando el programa informático GraphPad Prism v6.0, con un intervalo de confianza del 95% y una configuración para normalizar curvas de dosis- respuesta.
Análisis de toxicidad de compuestos Inhibition values were calculated using the average of 3 independent experiments (n=6) using GraphPad Prism v6.0 software, with a 95% confidence interval and a setting to normalize dose-response curves. Compound toxicity analysis
Se emplearon células Hela (2x104) tratadas con diferentes concentraciones: 10, 100 y 250 pM de cada compuesto durante 48 horas en placa de 96 pocilios. Tras este periodo, se lavaron las células con medio de cultivo y se midió la viabilidad mediante el método colorimétrico (Ensayo de proliferación celular no radiactivo Cell Titer 96 AQueous, Promega). Hela cells (2x10 4 ) treated with different concentrations were used: 10, 100 and 250 pM of each compound for 48 hours in a 96-well plate. After this period, the cells were washed with culture medium and viability was measured using the colorimetric method (Cell Titer 96 AQueous non-radioactive cell proliferation assay, Promega).
En resumen, 20 pL de la solución combinada de MTS/PMS (reactivo) se mezclaron con 100 pL de células en medio de cultivo y se incubaron adicionalmente durante 2 horas a 37 ºC en una atmósfera humidificada de CO2 al 5%. La absorbancia se registró a 490 nm utilizando un lector de placas ELISA. La viabilidad celular se calculó como el porcentaje de absorbancia en las células con el compuesto en relación con las células no tratadas. Briefly, 20 pL of the combined MTS/PMS solution (reagent) was mixed with 100 pL of cells in culture medium and further incubated for 2 h at 37 °C in a humidified 5% CO 2 atmosphere. Absorbance was recorded at 490 nm using an ELISA plate reader. Cell viability was calculated as the percentage of absorbance in cells with the compound relative to untreated cells.
Resultados
aCI50: Concentración inhibitoria 50% o requerida para reducir la infección viral en un 50% en los pseudotipos indicados en cada caso. bCCI50: Concentración citotóxica 50% o requerida para reducir el crecimiento celular en un 50% en el tipo de células indicados en cada caso CIS: índice de selectividad (CC50/CI50) Results at IC 50 : Inhibitory concentration 50% or required to reduce viral infection by 50% in the pseudotypes indicated in each case. b CCI 50 : Cytotoxic concentration 50% or required to reduce cell growth by 50% in the type of cells indicated in each case C IS: selectivity index (CC50/IC 50 )
Elemolo 3: Estudio de la actividad antiviral de los compuestos de fórmula (I) frente al virus de la oeste porcina africana (VPPA) Element 3: Study of the antiviral activity of the compounds of formula (I) against the West African swine virus (ASPV)
Materiales y Métodos
Tratamiento e Infección Materials and methods Treatment and Infection
Para la realización de los diversos ensayos, las infecciones con VPPA se realizaron sobre monocapas de células Vero (ATCC-CCL-81 ; fibroblastos renales) a una multiplicidad de infección (mdi) de 5 ufp/célula del aislado de VPPA Ba71 V no patógeno adaptado a células Vero (Enjuanes, L., Carrascosa, A.L., Moreno, M.A., Vinuela, E., 1976. Titration of African swine fever (ASF) virus. J Gen Virol 32, 471-477) o con los virus recombinantes fluorescentes para citometría de flujo como se describe más abajo. To carry out the various assays, infections with ASFV were carried out on monolayers of Vero cells (ATCC-CCL-81; renal fibroblasts) at a multiplicity of infection (mdi) of 5 pfu/cell of the non-pathogenic ASFV Ba71 V isolate. adapted to Vero cells (Enjuanes, L., Carrascosa, A.L., Moreno, M.A., Vinuela, E., 1976. Titration of African swine fever (ASF) virus. J Gen Virol 32, 471-477) or with fluorescent recombinant viruses for flow cytometry as described below.
Los inóculos víricos se añadieron siempre sobre el mínimo volumen necesario para cubrir el tapiz celular, tras el tratamiento previo de las células con los compuestos durante una hora a 37ºC . Transcurrido el tiempo de pre-tratamiento, se permitió la absorción del inóculo viral durante 60 minutos a 37 °C y por último, se retiró el medio junto con el inóculo viral y se reemplazó por medio de cultivo Eagle modificado por Dulbecco (DMEM) suplementado con 2% suero fetal bovino (SBF) en combinación con los distintos compuestos para proseguir con el tratamiento hasta alcanzar las 16 horas post infección (hpi). The viral inocula were always added to the minimum volume necessary to cover the cell mat, after pretreatment of the cells with the compounds for one hour at 37ºC . After the pre-treatment time had elapsed, the viral inoculum was allowed to absorb for 60 minutes at 37 °C and finally, the medium was removed along with the viral inoculum and replaced with supplemented Dulbecco's modified Eagle culture medium (DMEM). with 2% fetal bovine serum (FBS) in combination with the different compounds to continue the treatment until reaching 16 hours post infection (hpi).
Todos los compuestos se diluyeron inicialmente en dimetilsulfóxido (DMSO) a una concentración de 50 mM. A continuación, se diluyeron en medio DMEM al 2% en suero fetal bovino (FBS) hasta alcanzar la concentración requerida en cada caso. All compounds were initially diluted in dimethyl sulfoxide (DMSO) to a concentration of 50 mM. Next, they were diluted in 2% DMEM medium in fetal bovine serum (FBS) until reaching the concentration required in each case.
Ensayos de Cltotoxlcldad Chlototoxicity assays
La viabilidad celular en presencia de los distintos compuestos a diversas concentraciones, fue determinada a las 24 horas mediante el kit comercial “Cell titer 96 Aqueous Non-Radioactive Cell Proliferation Assay” (Promega) mediante la lectura de los valores de absorbencia a 490 nm, siguiendo las instrucciones del fabricante. Aquellas concentraciones que permitieron una viabilidad celular igual o superior al 80% fueron consideradas como no citotóxicas. Este sistema está basado en la cuantificación colorimétrica de la reducción del tetrazolio en formazán por la actividad de enzimas deshidrogenasas de células metabólicamente activas. Cell viability in the presence of the different compounds at various concentrations was determined after 24 hours using the commercial kit “Cell titer 96 Aqueous Non-Radioactive Cell Proliferation Assay” (Promega) by reading the absorbance values at 490 nm. following the manufacturer's instructions. Those concentrations that allowed cell viability equal to or greater than 80% were considered non-cytotoxic. This system is based on the colorimetric quantification of the reduction of tetrazolium to formazan by the activity of dehydrogenase enzymes of metabolically active cells.
Detección y cuantificación del DNA viral Detection and quantification of viral DNA
Posteriormente a la infección y el tratamiento con los distintos compuestos, se realizó la extracción de las células y la posterior obtención y purificación de ADN con el kit “Dneasy blood and tissue kit" (Quiagen). Las concentraciones obtenidas se cuantificaron en el
espectrofotómetro “UV-Vis Nanodrop ND- 1000” (Fisher Termo Scientific). After the infection and treatment with the different compounds, the cells were extracted and the subsequent DNA obtained and purified with the “Dneasy blood and tissue kit" (Qiagen). The concentrations obtained were quantified in the “UV-Vis Nanodrop ND- 1000” spectrophotometer (Fisher Termo Scientific).
Para el análisis de la replicación viral se realizó una PCR cuantitativa a tiempo real (qPCR) mediante sondas de hibridación fluorescentes dirigidas a amplificar la región del gen viral p72 como se ha descrito previamente (King, D. P.; Reid, S. M.; Hutchings, G. H.; Grierson, S. S.; Wilkinson, P. J.; Dixon, L. K.; Bastos, A. D.; Drew, T. W. Development of a TaqMan PCR assay with internal amplification control for the detection of African swine fever virus. J Virol Methods 2003, 107, 53-61). Para ello se analizaron las muestras de ADN purificadas previamente, utilizando como control negativo agua, como control positivo las muestras tratadas con DMSO y como curva patrón, 5 diluciones seriadas de ADN viral purificado de concentración conocida. La mezcla para la amplificación se compuso de 150 ng de ADN de cada una de las muestras, la sonda Taqman (Roche), los cebadores OE3F y OE4R cuya secuencia aparece descrita por King et al., obtenidos de Fisher Scientific y “Premix Ex Taq (2X)” (Takara). To analyze viral replication, real-time quantitative PCR (qPCR) was performed using fluorescent hybridization probes aimed at amplifying the region of the viral p72 gene as previously described (King, D. P.; Reid, S. M.; Hutchings, G. H.; Grierson, S. S.; Wilkinson, P. J.; Dixon, L. K.; Drew, T. W. Development of a TaqMan PCR assay with internal amplification control for the detection of African swine fever virus. To this end, the previously purified DNA samples were analyzed, using water as a negative control, samples treated with DMSO as a positive control, and 5 serial dilutions of purified viral DNA of known concentration as a standard curve. The mixture for amplification was composed of 150 ng of DNA from each of the samples, the Taqman probe (Roche), the primers OE3F and OE4R whose sequence is described by King et al., obtained from Fisher Scientific and “Premix Ex Taq (2X)” (Takara).
El proceso de amplificación y cuantificación se llevó a cabo en un termociclador “ABI 7500 Fast Real-Time PCR System” (Applied Biosystems) con los siguientes parámetros: 1 ciclo de 94ºC durante 10 minutos, 45 ciclos de 94 ºC durante 15 segundos y 58ºC durante 60 segundos. The amplification and quantification process was carried out in an “ABI 7500 Fast Real-Time PCR System” thermocycler (Applied Biosystems) with the following parameters: 1 cycle of 94 º C for 10 minutes, 45 cycles of 94 º C for 15 seconds and 58 º C for 60 seconds.
Cltometría de flujo Flow chlorometry
Para el análisis por citometría de flujo sin el mareaje con anticuerpos, las células fueron infectadas con los virus recombinantes realizados en el laboratorio BPP30GFP o B54GFP, descritos en Barrado-Gil, L; Galindo, I.; Martinez-Alonso, D.; Viedma, S.; Alonso, C. The ubiquitin-proteasome system is required for African swine fever replication. PLoS One 2017, 12, e018974 y en Hemaez, B.; Escribano, J. M.; Alonso, C. Visualization of the African swine fever virus infection in living cells by incorporation into the virus particle of green fluorescent protein-p54 membrane protein chimera. Virology 2006, 350, 1-14, respectivamente. Transcurridas 16 hpi las células se levantaron con tripsina y se centrifugaron a 2500 rpm durante 5 minutos. Los sobrenadantes se desecharon y las células se resuspendieron en tampón FACS (del inglés, “fluorescence- activated cell sorter’). Finalmente se analizaron 10 000 células por experimento en un dtómetro de flujo “FACS Canto II" (BD Science) para determinar el porcentaje de células infectadas. El porcentaje obtenido tras el tratamiento con los compuestos se normalizó con los valores de los controles. Una vez obtenidos los resultados con las
concentraciones óptimas, se calcularon las CI50 empleando diferentes concentraciones de compuestos. For analysis by flow cytometry without labeling with antibodies, the cells were infected with the recombinant viruses made in the laboratory BPP30GFP or B54GFP, described in Barrado-Gil, L; Galindo, I.; Martinez-Alonso, D.; Viedma, S.; Alonso, C. The ubiquitin-proteasome system is required for African swine fever replication. PLoS One 2017, 12, e018974 and in Hemaez, B.; Scribe, JM; Alonso, C. Visualization of the African swine fever virus infection in living cells by incorporation into the virus particle of green fluorescent protein-p54 membrane protein chimera. Virology 2006, 350, 1-14, respectively. After 16 hpi, the cells were lifted with trypsin and centrifuged at 2500 rpm for 5 minutes. The supernatants were discarded and the cells were resuspended in FACS (fluorescence-activated cell sorter) buffer. Finally, 10,000 cells per experiment were analyzed in a “FACS Canto II" flow meter (BD Science) to determine the percentage of infected cells. The percentage obtained after treatment with the compounds was normalized with the values of the controls. Once obtained the results with the optimal concentrations, IC50s were calculated using different concentrations of compounds.
Análisis estadístico Statistic analysis
Todos los datos se obtuvieron por la realización por triplicado de los experimentos y la normalización de las medidas a los valores control. El análisis estadístico de la varianza entre datos se realizó con el software Graph Pad Prism 6, aplicando el test de Bonferroni en las comparaciones múltiples. All data were obtained by performing the experiments in triplicate and normalizing the measurements to control values. The statistical analysis of the variance between data was performed with the Graph Pad Prism 6 software, applying the Bonferroni test in multiple comparisons.
Resultados
aCI50: Concentración inhibitoria 50% o requerida para reducir la infección viral en un 50%. bCCI50: Concentración citotóxica 50% o requerida para reducir el crecimiento celular en un 50%. Results at IC 50 : Inhibitory concentration 50% or required to reduce viral infection by 50%. b CCI 50 : Cytotoxic concentration 50% or required to reduce cell growth by 50%.
CIS: índice de selectividad (CC50/CI50).
C IS: selectivity index (CC50/IC50).
Claims
1. Compuesto de fórmula (I) o cualquiera de sus isómeros o sus sales farmacéuticamente aceptables
( ) donde: R1 es un grupo seleccionado de y -NH(CH2)nAr,
siendo: Bn un grupo bencilo n un número entero comprendido entre O y 2 y 1. Compound of formula (I) or any of its isomers or its pharmaceutically acceptable salts ( ) where: R 1 is a group selected from and -NH(CH 2 )nAr, where: Bn is a benzyl group n is an integer between O and 2 and
Ar un grupo fenilo sin sustituir o un grupo fenilo sustituido con un halógeno o con el heterociclo
; Ar is an unsubstituted phenyl group or a phenyl group substituted with a halogen or with the heterocycle ;
R2, R3 y R4 se seleccionan independientemente de -H, -OH, -O-alquilo C1-C3 y R 2 , R 3 and R 4 are independently selected from -H, -OH, -O-C1-C3 alkyl and
, donde al menos dos radicales seleccionados de R2, R3 y R4 son -H;
y con la condición de que si R2, R3 y R4 son H,-R1 es
, where at least two radicals selected from R 2 , R 3 and R 4 are -H; and with the condition that if R 2 , R 3 and R 4 are H,-R 1 is
20
y si uno de los radicales seleccionados de R2, R3 y R4 es -OH, R1 es
y donde el compuesto de formula (I) no es: o. twenty and if one of the radicals selected from R 2 , R 3 and R 4 is -OH, R 1 is and where the compound of formula (I) is not: o.
Cl para su uso como medicamento. Cl for use as a medicine.
2. Compuesto para su uso, según reivindicación 1 , donde R1 es un grupo seleccionado de y -NH(CH2)nAr, donde n es 0 ó 2.
2. Compound for use, according to claim 1, where R 1 is a group selected from and -NH (CH2) nAr, where n is 0 or 2.
3. Compuesto para su uso, según reivindicación 1 , donde R1 es un grupo - NH(CH2)nAr donde n es 0 y Ar es un fenilo sin sustituir o sustituido por un cloro. 3. Compound for use, according to claim 1, where R 1 is a group - NH (CH2) nAr where n is 0 and Ar is an unsubstituted phenyl or substituted by a chlorine.
4. Compuesto para su uso, según reivindicación 1 , donde R1 es un grupo - NH(CH2)nAr donde n es 2 y Ar es un fenilo sin sustituir. 4. Compound for use, according to claim 1, where R 1 is a group - NH (CH 2 )nAr where n is 2 and Ar is an unsubstituted phenyl.
5. Compuesto para su uso, según cualquiera de las reivindicaciones 1 a 4, donde un radical seleccionado de R2, R3 y R4 es
5. Compound for use according to any of claims 1 to 4, wherein a radical selected from R 2 , R 3 and R 4 is
6. Compuesto para su uso, según cualquiera de las reivindicaciones 1 a 4, donde R2, R3 y R4 son H. 6. Compound for use according to any of claims 1 to 4, where R 2 , R 3 and R 4 are H.
7. Compuesto para su uso, según cualquiera de las reivindicaciones 1 a 4, donde uno de R2, R3 y R4 se seleccionan independientemente de -OH y -OCH3. 7. Compound for use according to any of claims 1 to 4, wherein one of R 2 , R 3 and R 4 are independently selected from -OH and -OCH 3 .
8. Compuesto para su uso, según reivindicación 1 , donde R1 es
uno de R2, R3 y R4 es -OCHa.
8. Compound for use, according to claim 1, where R 1 is one of R 2 , R 3 and R 4 is -OCHa.
9. Compuesto para su uso, según reivindicación 1, donde el compuesto es seleccionado de la siguiente lista: N-fenil-5-(2-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida N-fenil-5-(3-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida N-fenil-5-(4-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida N-(3-clorofenil)-5-feniltiofen-2-carboxamida N-(2-(1-bencilpiperidin-4-il)etil)-5-feniltiofen-2-carboxamida N-(2-(1-bencilpiperidin-4-il)etil)-5-(2-hidroxifenil)tiofen-2-carboxamida N-(4-clorofenil)-5-(2-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida N-(3-clorofenil)-5-(2-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida N-fenetil-5-(2-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida N-(4-morfolinofenil)-5-(2-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida N-(2-(1-bencilpiperidin-4-il)etil)-5-(2-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida N-(3-clorofenil)-5-(4-hidroxifenil)tiofen-2-carboxamida (N-(2-(1-bencilpiperidin-4-il)etil)-5-(4-hidroxifenil)tiofen-2-carboxamida N-(3-clorofenil)-5-(3-hidroxifenil)tiofen-2-carboxamida N-(2-(1-bencilpiperidin-4-il)etil)-5-(3-hidroxifenil)tiofen-2-carboxamida N-(4-clorofenil)-5-(3-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida N-(3-clorofenil)-5-(3-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida N-fenetil-5-(3-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida N-(2-(1-bencilpiperidin-4-il)etil)-5-(4-metoxifenil)tiofen-2-carboxamida N-(2-(1-bencilpiperidin-4-il)etil)-5-(3-metoxifenil)tiofen-2-carboxamida N-(4-clorofenil)-5-(4-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida N-(3-clorofenil)-5-(4-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida 9. Compound for use, according to claim 1, wherein the compound is selected from the following list: N-phenyl-5-(2-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide N-phenyl-5- (3-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide N-phenyl-5-(4-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide N-(3-chlorophenyl)-5- phenylthiophen-2-carboxamide N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-phenylthiophen-2-carboxamide N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-(2 -hydroxyphenyl)thiophen-2-carboxamide N-(4-chlorophenyl)-5-(2-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide N-(3-chlorophenyl)-5-(2-(piperidin) -4-yloxy)phenyl)thiophen-2-carboxamide N-phenethyl-5-(2-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide N-(4-morpholinophenyl)-5-(2-(piperidin) -4-yloxy)phenyl)thiophen-2-carboxamide N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-(2-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide N- (3-chlorophenyl)-5-(4-hydroxyphenyl)thiophen-2-carboxamide (N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-(4-hydroxyphenyl)thiophen-2-carboxamide N- (3-chlorophenyl)-5-(3-hydroxyphenyl)thiophen-2-carboxamide N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-(3-hydroxyphenyl)thiophen-2-carboxamide N-( 4-chlorophenyl)-5-(3-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide N-(3-chlorophenyl)-5-(3-(piperidin-4-yloxy)phenyl)thiophen-2- carboxamide N-phenethyl-5-(3-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-(4-methoxyphenyl)thiophen- 2-carboxamide N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-(3-methoxyphenyl)thiophen-2-carboxamide N-(4-chlorophenyl)-5-(4-(piperidin-4- yloxy)phenyl)thiophen-2-carboxamide N-(3-chlorophenyl)-5-(4-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide
10. Compuesto de fórmula (I) o cualquiera de sus isómeros o sus sales farmacéuticamente aceptables
donde R1, R2, R3 y R4 son como se ha definido en cualquiera de las reivindicaciones 1 a 9, con la condición de que si R2, R3 y R4 son H,-R1 es
H O y si uno de los radicales seleccionados de R2, R3 y R4 es -OH, R1 es
para su uso en el tratamiento y/o prevención de enfermedades
víricas. 10. Compound of formula (I) or any of its isomers or its pharmaceutically acceptable salts where R 1 , R 2 , R 3 and R 4 are as defined in any of claims 1 to 9, with the proviso that if R 2 , R 3 and R 4 are H, -R 1 is HO and if one of the radicals selected from R 2 , R 3 and R 4 is -OH, R 1 is for use in the treatment and/or prevention of diseases viral.
11 . Compuesto para su uso según reivindicación 10 donde las enfermedades víricas se seleccionan de enfermedad del Ébola y la peste porcina africana. eleven . Compound for use according to claim 10 wherein the viral diseases are selected from Ebola disease and African swine fever.
12. Composición farmacéutica que comprende al menos un compuesto de fórmula (I) según se ha definido en la reivindicación 10 junto con un vehículo o excipiente farmacéuticamente aceptable para su uso en el tratamiento de enfermedades víricas. 12. Pharmaceutical composition comprising at least one compound of formula (I) as defined in claim 10 together with a pharmaceutically acceptable vehicle or excipient for use in the treatment of viral diseases.
13. Composición farmacéutica para su uso, según reivindicación 12, donde las enfermedades víricas se seleccionan de enfermedad del Ébola y la peste porcina africana 13. Pharmaceutical composition for use, according to claim 12, wherein the viral diseases are selected from Ebola disease and African swine fever.
14. Compuesto de fórmula (I) o cualquiera de sus isómeros o sus sales farmacéuticamente aceptables:
donde: 14. Compound of formula (I) or any of its isomers or its pharmaceutically acceptable salts: where:
R1 es un grupo seleccionado
-NH(CH2)nAr,
siendo Bn un grupo bencilo n un número entero comprendido entre 0 y 2 y R 1 is a selected group -NH(CH 2 )nAr, Bn being a benzyl group n an integer between 0 and 2 and
Ar un grupo fenilo (Ph) sin sustituir o un grupo fenilo sustituido con un halógeno o con el heterociclo
Ar is an unsubstituted phenyl (Ph) group or a phenyl group substituted with a halogen or with the heterocycle
R2, R3 y R4 se seleccionan independientemente de -H, -OH, -OCH3 y
donde al menos dos de R2, R3 y R4 son -H; y con la condición de que si R2, R3 y R4son iguales a -H, R1 es
H y si uno de R2, R3 y R4 es -OH, R1 es
y donde el compuesto no es el siguiente:
R 2 , R 3 and R 4 are independently selected from -H, -OH, -OCH3 and where at least two of R 2 , R 3 and R 4 are -H; and with the condition that if R 2 , R 3 and R 4 are equal to -H, R 1 is H and if one of R 2 , R 3 and R 4 is -OH, R 1 is and where the compound is not the following:
15. Compuesto, según reivindicación 14, donde R1 es un grupo seleccionado de y - NH(CH2)nAr, donde n es 0 ó 2. 15. Compound according to claim 14, where R 1 is a group selected from y - NH (CH 2 )nAr, where n is 0 or 2.
16. Compuesto, según reivindicación 14, donde R1 es un grupo -NH(CH2)nAr donde n es 0 y Ar es un fenilo sin sustituir o sustituido por un cloro. 16. Compound, according to claim 14, where R 1 is a group -NH (CH 2 )nAr where n is 0 and Ar is an unsubstituted phenyl or substituted by a chlorine.
17. Compuesto, según reivindicación 14, donde R1 es un grupo -NH(CH2)nAr donde n es 2 y Ar es un fenilo sin sustituir. 17. Compound according to claim 14, where R 1 is a group -NH (CH 2 )nAr where n is 2 and Ar is an unsubstituted phenyl.
18. Compuesto, según cualquiera de las reivindicaciones 14 a 17, donde R2o R3 o 18. Compound according to any of claims 14 to 17, where R 2 or R 3 or
R4 es
R 4 is
19. Compuesto, según cualquiera de las reivindicaciones 14 a 17, donde R2, R3 y R4 son -H. 19. Compound according to any of claims 14 to 17, where R 2 , R 3 and R 4 are -H.
20. Compuesto, según cualquiera de las reivindicaciones 14 a 17, donde uno de R2, R3 y R4 se seleccionan independientemente de OH y OCH3. 20. Compound according to any of claims 14 to 17, wherein one of R 2 , R 3 and R 4 are independently selected from OH and OCH 3 .
21. Compuesto, según reivindicación 14, donde R1 es y uno de21. Compound, according to claim 14, where R 1 is and one of
R2, R3 y R4 es -OCH3.
R 2 , R 3 and R 4 is -OCH 3 .
22. Compuesto, según reivindicación 14, donde el compuesto es seleccionado de la siguiente lista: 22. Compound, according to claim 14, wherein the compound is selected from the following list:
N-fenil-5-(2-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida N-phenyl-5-(2-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide
N-fenil-5-(3-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida N-phenyl-5-(3-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide
N-fenil-5-(4-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida N-phenyl-5-(4-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide
N-(2-(1-bencilpiperidin-4-il)etil)-5-feniltiofen-2-carboxamida N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-phenylthiophen-2-carboxamide
N-(2-(1-bencilpiperidin-4-il)etil)-5-(2-hidroxifenil)tiofen-2-carboxamida N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-(2-hydroxyphenyl)thiophen-2-carboxamide
N-(4-clorofenil)-5-(2-(piperidin-4-iloxi)fenil)tiofen-2-cartx)xamida N-(4-chlorophenyl)-5-(2-(piperidin-4-yloxy)phenyl)thiophen-2-cartx)xamide
N-(3-clorofenil)-5-(2-(piperidin-4-iloxi)fenil)tiofen-2-cartx)xamida N-(3-chlorophenyl)-5-(2-(piperidin-4-yloxy)phenyl)thiophen-2-cartx)xamide
N-fenetil-5-(2-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida N-phenethyl-5-(2-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide
N-(4-morfolinofenil)-5-(2-(piperidin-4-iloxi)fenil)tiofen-2-cartx)xamida N-(4-morpholinophenyl)-5-(2-(piperidin-4-yloxy)phenyl)thiophen-2-cartx)xamide
N-(2-(1-bencilpiperidin-4-il)etil)-5-(2-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-(2-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide
N-(3-clorofenil)-5-(4-hidroxifenil)tiofen-2-cartx)xamida (N-(2-(1-bencilpiperidin-4-il)etil)-5-(4-hidroxifenil)tiofen-2-cartx)xamida N-(3-dorofenil)-5-(3-hidroxifenil)tiofen-2-cartx)xamida N-(3-chlorophenyl)-5-(4-hydroxyphenyl)thiophen-2-cartx)xamide (N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-(4-hydroxyphenyl)thiophen-2 -cartx)xamide N-(3-dorophenyl)-5-(3-hydroxyphenyl)thiophen-2-cartx)xamide
N-(2-(1-bencilpiperidin-4-il)etil)-5-(3-hidroxifenil)tiofen-2-carboxamida N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-(3-hydroxyphenyl)thiophen-2-carboxamide
N-(4-clorofenil)-5-(3-(piperidin-4-iloxi)fenil)tiofen-2-cartx)xamida N-(4-chlorophenyl)-5-(3-(piperidin-4-yloxy)phenyl)thiophen-2-cartx)xamide
N-(3-clorofenil)-5-(3-(piperidin-4-iloxi)fenil)tiofen-2-cartx)xamida N-(3-chlorophenyl)-5-(3-(piperidin-4-yloxy)phenyl)thiophen-2-cartx)xamide
N-fenetil-5-(3-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida N-phenethyl-5-(3-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide
N-(2-(1-bencilpiperidin-4-il)etil)-5-(4-metoxifenil)tiofen-2-carboxamida N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-(4-methoxyphenyl)thiophen-2-carboxamide
N-(2-(1-bencilpiperidin-4-il)etil)-5-(3-metoxifenil)tiofen-2-carboxamida N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-(3-methoxyphenyl)thiophen-2-carboxamide
N-(4-clorofenil)-5-(4-(piperidin-4-iloxi)fenil)tiofen-2-cartx)xamida N-(3-clorofenil)-5-(4-(piperidin-4-iloxi)fenil)tiofen-2-cartx)xamida. N-(4-chlorophenyl)-5-(4-(piperidin-4-yloxy)phenyl)thiophen-2-cartx)xamide N-(3-chlorophenyl)-5-(4-(piperidin-4-yloxy)phenyl )thiophen-2-cartx)xamide.
23. Composición farmacéutica que comprende al menos un compuesto de fórmula
(I) definido en cualquiera de las reivindicaciones 14 a 22 junto con un vehículo o excipiente farmacéuticamente aceptable.
23. Pharmaceutical composition comprising at least one compound of formula (I) defined in any of claims 14 to 22 together with a pharmaceutically acceptable vehicle or excipient.
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| WO2019036562A1 (en) * | 2017-08-18 | 2019-02-21 | Saint Louis University | Err inverse agonists |
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Non-Patent Citations (8)
| Title |
|---|
| ASCUAL MARÍA JOSÉ; MERWAISS FERNANDO; LEAL EMILSE; QUINTANA MARÍA EUGENIA; CAPOZZO ALEJANDRA V; CAVASOTTO CLAUDIO N; BOLLINI MARIE: "Structure-based drug design for envelope protein E2 uncovers a new class of bovine viral diarrhea inhibitors that block virus entry", ANTIVIRAL RESEARCH, vol. 149, 1 January 2018 (2018-01-01), NL , pages 179 - 190, XP085315569, ISSN: 0166-3542, DOI: 10.1016/j.antiviral.2017.10.010 * |
| DATABASE REGISTRY 1 July 2001 (2001-07-01), ANONYMOUS: "2-Thiophenecarboxamide, N-(4-chlorophenyl)-5-(4-methoxyphenyl)- (CA INDEX NAME)", XP093176976, retrieved from STN Database accession no. 344263-75-8 * |
| DATABASE REGISTRY 22 February 2007 (2007-02-22), ANONYMOUS: "2-Thiophenecarboxamide, 5-(4-ethoxyphenyl)-N-(phenylmethyl)- (CA INDEX NAME)", XP093176970, retrieved from STN Database accession no. 922436-24-6 * |
| DATABASE REGISTRY 27 February 2007 (2007-02-27), ANONYMOUS: "2-Thiophenecarboxamide, 5-(4-ethoxyphenyl)-N-[(3- fluorophenyl)methyl]- (CA INDEX NAME)", XP093176961, Database accession no. 923517-02-6 * |
| DATABASE REGISTRY 27 February 2007 (2007-02-27), ANONYMOUS: "2-Thiophenecarboxamide, N-[(2-chlorophenyl)methyl]-5-(4- ethoxyphenyl)- (CA INDEX NAME)", XP093176968, retrieved from STN Database accession no. 923543-36-6 * |
| DATABASE REGISTRY 27 February 2007 (2007-02-27), ANONYMOUS: "2-Thiophenecarboxamide, N-[(4-chlorophenyl)methyl]-5-(4- ethoxyphenyl)- (CA INDEX NAME)", XP093176963, retrieved from STN Database accession no. 923537-15-9 * |
| DATABASE REGISTRY 27 February 2007 (2007-02-27), AURORA FINE CHEMICALS: "2-Thiophenecarboxamide, N-[2-(3-chlorophenyl)ethyl]-5-(4- ethoxyphenyl)- (CA INDEX NAME)", XP093176959, retrieved from STN Database accession no. 923531-76-4 * |
| DATABASE REGISTRY 6 July 2006 (2006-07-06), ANONYMOUS: "2-Thiophenecarboxamide, N-(3-chlorophenyl)-5-phenyl- (CA INDEX NAME) ", XP093176973, retrieved from STN Database accession no. 890826-75-2. * |
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