DESCRIPCIÓNDESCRIPTION
Compuestos derivados de tiofeno como agentes antiviralesThiophene-derived compounds 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 relates 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 INVENCIÓNBACKGROUND 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 especificos es muy grande (Malvy, D.; McElroy, A. K.; de Clerck, H.; Gunther, S.; van Griensven, J. Ebola virus disease.Lancet2019,393,936-948). 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.; Andrés, G. A Proteomic atlas of the African swine fever virus partióle.J Virol2018,92,e01293-18).We are currently witnessing an increase in emerging and re-emerging viral diseases worldwide. In fact, infectious diseases are one of the greatest concerns for both human and veterinary public health. Such is the case of Ebola virus disease. It is a very serious and contagious disease, causing hemorrhagic fever and with a high mortality rate in humans, which can reach 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.Lancet2019,393,936-948). 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 impact. The African swine fever virus (ASFV) causes a hemorrhagic disease with high mortality in domestic pigs and wild boars (Alejo, A.; Matamoros, T.; Guerra, M.; Andrés, G. A Proteomic atlas of the African swine fever virus partióle.J Virol2018,92,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 INVENCIÓNDESCRIPTION OF THE INVENTION
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.The present invention presents a family of compounds that possess antiviral activity in the micromolar order, resulting 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:In a first aspect, the present invention relates to a compound of formula (I) or any of its isomers or its pharmaceutically acceptable salts:
donde:where:
R1 es un grupo seleccionadoR1 is a selected group
siendo Bn un grupo benciloBn being a benzyl group
n un número entero comprendido entre 0 y 2 yn an integer between 0 and 2 and
Ar un grupo fenilo (Ph) sin sustituir o un grupo fenilo sustituido con un halógenoAr an unsubstituted phenyl group (Ph) or a phenyl group substituted with a halogen
o con el heterocicloor with the heterocycle
R2, R3 y R4 se seleccionan independientemente de -H, -OH, -O-alquilo C<1>-C<3>yR2, R3 and R4 are independently selected from -H, -OH, -O-C<1>-C<3>alkyl and
, donde al menos dos radicales seleccionados de R2, R3 y R4 son H;, where at least two radicals selected from R2, R3 and R4 are H;
y con la condición de que si R2, R3 y R4 son -and provided that if R2, R3 and R4 are -
para su uso como medicamento.for use as a medicine.
En una realización preferida, R1 es un grupo seleccionado de y -NH(CH<2>)nAr, donde n es 0 ó 2.In a preferred embodiment, R1 is a group selected from -NH(CH<2>)nAr, where n is 0 or 2.
En otra realización preferida, R1 es un grupo -NH(CH<2>)nAr, donde n es 0 ó 2.In another preferred embodiment, R1 is a group -NH(CH<2>)nAr, where n is 0 or 2.
En otra realización preferida, R1 es un grupo -NH(CH<2>)nAr donde n es 0 y Ar es un fenilo sin sustituir o sustituido por un cloro.In another preferred embodiment, R1 is a group -NH(CH<2>)nAr where n is 0 and Ar is an unsubstituted phenyl or substituted by a chlorine.
En otra realización preferida, R1 es un grupo -NH(CH<2>)nAr donde n es 2 y Ar es un fenilo sin sustituir.In another preferred embodiment, R1 is a group -NH(CH<2>)nAr 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 independently selected from H,
OH, OCH<3>yOH, OCH<3>and
En una realización preferida, R2 esIn a preferred embodiment, R2 is
En una realización preferida, R3 esIn a preferred embodiment, R3 is
En una realización preferida, R4 esIn a preferred embodiment, R4 is
En otra realización preferida, R2, R3 y R4 son -H.In another preferred embodiment, R2, R3 and R4 are -H.
En otra realización preferida, R2, R3 y R4 se seleccionan independientemente de -OH y O-alquilo C<1>-C<3>, preferiblemente -OCH<3>, siendo los otros dos sustituyentes H.In another preferred embodiment, R2, R3 and R4 are independently selected from -OH and O-C<1>-C<3>alkyl, preferably -OCH<3>, with the other two substituents being H.
En otra realización preferida, R1 es y uno de R2, R3 y R4 es -O-alquilo C<1>-C<3>, preferiblemente -OCH<3>.In another preferred embodiment, R1 is and one of R2, R3 and R4 is -O-C<1>-C<3>alkyl, preferably -OCH<3>.
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: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-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-(3-clorofenil)-5-feniltiofen-2-carboxamida (1)N-(3-chlorophenyl)-5-phenylthiophen-2-carboxamide (1)
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-(3-clorofenil)-5-(4-hidroxifenil)tiofen-2-carboxamida (14)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-(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)
Otro aspecto de la invención se refiere a los compuestos de fórmula general (I) como se han descrito en el primer aspecto de la invención para su uso en el tratamiento y/o prevención de enfermedades víricas, preferiblemente la enfermedad del Ébola y de la peste porcina africana.Another aspect of the invention relates to the compounds of general formula (I) as described in the first aspect of the invention for use in the treatment and/or prevention of viral diseases, preferably Ebola disease and African swine fever.
La presente invención también se refiere a una composición farmacéutica que comprende al menos un compuesto de fórmula (I) definido anteriormente 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) defined above together with a pharmaceutically acceptable carrier or excipient 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.The compound must be in the therapeutically effective amount for use in the treatment and/or prevention of viral diseases.
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) to be treated, the severity of the disease suffered by said individual, the form of administration chosen, etc. For this reason, the doses mentioned in this invention should be considered only as guides for the person skilled in the art, and the latter 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:The present invention also relates to a compound of formula (I) or any of its isomers or its pharmaceutically acceptable salts:
donde:where:
R1 es un grupo seleccionado de y -NH(CH<2>)nAr,R1 is a group selected from -NH(CH<2>)nAr,
siendo Bn un grupo benciloBn being a benzyl group
n un número entero comprendido entre 0 y 2 yn an integer between 0 and 2 and
Ar un grupo fenilo (Ph) sin sustituir o un grupo fenilo sustituido con un halógenoAr an unsubstituted phenyl group (Ph) or a phenyl group substituted with a halogen
o con el heterocicloor with the heterocycle
R2, R3 y R4 se seleccionan independientemente de -H, -OH, -O-Alquilo C<1>-C<3>yR2, R3 and R4 are independently selected from -H, -OH, -O-C<1>-C<3>Alkyl and
, donde al menos dos de R2, R3 y R4 son -H;, where at least two of R2, R3 and R4 are -H;
y con la condición de que si R2, R3 y R4 son iguales a -H, R1 esand with the condition that if R2, R3 and R4 are equal to -H, R1 is
y si uno de R2, R3 y R4 es -OH, R1 esand if one of R2, R3 and R4 is -OH, R1 is
y donde el compuesto no es el siguiente:and where the compound is not the following:
En una realización preferida, R1 es un grupo seleccionado de y -NH(CH<2>)nAr, donde n es 0 ó 2.In a preferred embodiment, R1 is a group selected from -NH(CH<2>)nAr, where n is 0 or 2.
En otra realización preferida, R1 es un grupo -NH(CH<2>)nAr, donde n es 0 ó 2.In another preferred embodiment, R1 is a group -NH(CH<2>)nAr, where n is 0 or 2.
En una realización preferida, R1 es un grupo -NH(CH<2>)nAr donde n es 0 y Ar es un fenilo sin sustituir o sustituido por un cloro.In a preferred embodiment, R1 is a group -NH(CH<2>)nAr where n is 0 and Ar is an unsubstituted phenyl or substituted by a chlorine.
En otra realización preferida, R1 es un grupo -NH(CH<2>)nAr donde n es 2 y Ar es un fenilo sin sustituir.In another preferred embodiment, R1 is a group -NH(CH<2>)nAr 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 independently selected from -H, -
En una realización preferida, R2 esIn a preferred embodiment, R2 is
En una realización preferida, R3 esIn a preferred embodiment, R3 is
En una realización preferida, R4 es .In a preferred embodiment, R4 is .
En otra realización preferida, R2, R3 y R4 son H-.In another preferred embodiment, R2, R3 and R4 are H-.
En otra realización preferida, R2, R3 y R4 se seleccionan independientemente de OH y O-alquilo C<1>-C<3>, preferiblemente -OCH<3>, siendo los otros dos sustituyentes H.In another preferred embodiment, R2, R3 and R4 are independently selected from OH and O-C<1>-C<3>alkyl, preferably -OCH<3>, with the other two substituents being H.
En otra realización preferida, R1 es y uno de R2, R3 y R4 es un O-alquilo C<1>-C<3>, preferiblemente -OCH<3>.In another preferred embodiment, R1 is and one of R2, R3 and R4 is an O-C<1>-C<3>alkyl, preferably -OCH<3>.
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 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-(3-clorofenil)-5-(4-hidroxifenil)tiofen-2-carboxamida (14)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-(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 relates to a pharmaceutical composition comprising at least one compound of formula (I) defined in the preceding aspect of the invention together with a pharmaceutically acceptable carrier 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, i.e. solvates of the compound of formula (I) which can be used in the manufacture of a medicament, and non-pharmaceutically acceptable 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. Solvates may 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 subcutaneously, intraperitoneally, intradermally, intramuscularly, intravenously, 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 herein 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 can form part of the same pharmaceutical composition or, alternatively, can be provided in the form of a separate composition for simultaneous or non-simultaneous administration with the pharmaceutical composition comprising a compound of formula (I), or a pharmaceutically acceptable salt or 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 which differ only in the presence of one or more isotopically enriched atoms. For example, compounds having such a structure, except for the substitution of a hydrogen by a deuterium or by tritium, or the substitution of a carbon by a 13C or 14C enriched carbon or a 15N enriched nitrogen, are within the scope of this invention.
En la presente invención, “-Bn” se refiere a un grupo bencilo, esto es: -CH<2>-Ph, siendoIn 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, cloro, bromo o yodo. En la presente invención, el halógeno proferido es el cloro.The term “halogen” refers in the present invention to fluorine, chlorine, bromine or iodine. In the present invention, the halogen referred to 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 aliphatic hydrocarbon chains, straight or branched, 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 reversing, alleviating, inhibiting the progress of, or preventing the disorder or condition to which such terms apply, or 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 pharmaceutical excipients, adjuvants or 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.Throughout the description and claims the word "comprises" and its variants are not intended to exclude other technical features, additives, components or steps. For those skilled in the art, other objects, advantages and features of the invention will become apparent 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.
EJEMPLOSEXAMPLES
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.The invention will now be illustrated by means of 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 AVIII 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 ^m, 4,6 mm x 50 mm); elución en gradiente de H<2>O/CH<3>CN de 100/0 a 0/100 durante 5 min; flujo 800 ^L/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 under an inert atmosphere. Reagents used for the reactions were purchased from Sigma Aldrich or Fluorochem. Microwave reactions were carried out with an InitiatorTM device (Biotage®). Melting points were recorded with a Büchi Melting point M-560 apparatus. 1H and 13C NMR spectra were recorded on a Bruker AVIII 300 MHz BACS60 or Bruker AV 500 MHz spectrometer located at the Research Support Center of the Complutense University of Madrid. 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). Pre-coated aluminum foils (ALUGRAM® Xtra SIL G/UV254) were used for thin layer chromatography. Column chromatographies were performed on silica gel 60 (Merk) manually or automatically using IsoleraOne equipment (Biotage®). The purity of the final compounds was verified to be >95% by HPLC analysis. HPLC conditions for assessing purity were as follows: Surveyor HPLC equipped with a Surveyor PDA plus UV-VIS detector; ZORBAX® SB-C18 column (3.5 ^m, 4.6 mm x 50 mm); Gradient elution of H<2>O/CH<3>CN from 100/0 to 0/100 over 5 min; flow rate 800 ^L/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 Mass Spectrometry Service of the Institute of General Organic Chemistry (IQOG-CSIC).
Síntesis de 5-fenilt¡ofen-2-carboxamidas (1-3) y 5-bromotiofen-2-carboxam¡das(4 8)Synthesis of 5-phenylthiophene-2-carboxamides (1-3) and 5-bromothiophene-2-carboxamides (4-8)
(4-8)(4-8)
Esquema 1. Procedimiento general de síntesis. Reactivos y condiciones: i) EDC, Et<3>N, HOBt, CH<2>CE 0 °C a 25 °C, 24h.Scheme 1. General synthesis procedure. Reagents and conditions: i) EDC, Et<3>N, HOBt, CH<2>CE 0 °C to 25 °C, 24h.
Procedimiento general: se añadió lentamente una solución de la amina correspondiente (1,02 equiv.) en CH<2>Ch a una solución del ácido 2-tiofeno-carboxílico adecuado (1 equiv.), 1-etil-3-(3-dimetilaminopropil)carbodiimida (EDC) (1 ó 2 equiv.) y Et<3>N (2 ó 3 equiv.) en CH<2>Ch a 0 °C. 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 H<2>O, disolución saturada de NH<4>Cl, y disolución saturada de cloruro sódico para finalmente secarla sobre Na<2>SO<4>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 CH<2>Ch was slowly added to a solution of the appropriate 2-thiophenecarboxylic acid (1 equiv.), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) (1 or 2 equiv.) and Et<3>N (2 or 3 equiv.) in CH<2>Ch at 0 °C. For some derivatives, 1-hydroxybenzotriazole (HOBt) (1 equiv.) was also added. After stirring the reaction mixture overnight at room temperature, the crude was washed with H<2>O, saturated NH<4>Cl solution, and saturated sodium chloride solution and finally dried over anhydrous Na<2>SO<4>. The desiccant was then filtered and the volatiles were evaporated to dryness in vacuo. 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-Clorofenil)-5-feniltiofen-2-carboxamida (1)N-(3-Chlorophenyl)-5-phenylthiophen-2-carboxamide (1)
Reactivos: Ácido 5-feniltiofen-2-carboxílico (1,5 mmol, 306 mg), 3-cloroanilina (1,53 mmol, 195 mg), EDC (3 mmol, 573 mg), Et<3>N (4,5 mmol, 626 pL) y CH<2>CI<2>(6,0 mL). Purificación: CH<2>Ch/MeOH (9:1). Rendimiento: 69 mg, 15%. Sólido beige. Pf: 155 - 157 °C. 1H RMN (300 MHz, DMSO-de) 5 10,39 (s, 1H), 8,04 (d,J= 4,0 Hz, 1H), 7,93 (t,J= 2,0 Hz, 1H), 7,73-7,81 (m, 2H), 7,69 (ddd,J= 8,3, 2,0, 1,0, 1H), 7,65 (d,J= 4,0 Hz, 1H), 7.51 - 7,37 (m, 4H), 7,18 (ddd,J= 8,0, 2,1, 0,9 Hz, 1H). 13C RMN (75 MHz, DMSO-de) 5 159,9, 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]+ calc. para C<17>H<13>CINOS 314,0401; encontrado 314,0400. Pureza (HPLC): 97,8 %.Reagents: 5-Phenylthiophene-2-carboxylic acid (1.5 mmol, 306 mg), 3-chloroaniline (1.53 mmol, 195 mg), EDC (3 mmol, 573 mg), Et<3>N (4.5 mmol, 626 pL) and CH<2>Cl<2>(6.0 mL). Purification: CH<2>Ch/MeOH (9:1). Yield: 69 mg, 15%. Beige solid. Mp: 155 - 157 °C. 1H NMR (300 MHz, DMSO-de) 5 10.39 (s, 1H), 8.04 (d,J= 4.0 Hz, 1H), 7.93 (t,J= 2.0 Hz, 1H), 7.73-7.81 (m, 2H), 7.69 (ddd,J= 8.3, 2.0, 1.0, 1H), 5 (d,J= 4.0 Hz, 1H), 7.51 - 7.37 (m, 4H), 7.18 (ddd,J= 8.0, 2.1, 0.9 Hz, 1H). 13C NMR (75 MHz, DMSO-de) 5 159.9, 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, 118.5. HRMS (ESI) m/z: [M H]+ calc. for C<17>H<13>CINOS 314.0401; found 314.0400. Purity (HPLC): 97.8%.
N-(2-(1-Bencilpiperidin-4-il)etil)-5-feniltiofen-2-carboxamida (2)N-(2-(1-Benzylpiperidin-4-yl)ethyl)-5-phenylthiophen-2-carboxamide (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), Et<3>N (4,5 mmol, 626 pL) y CH<2>Cl<2>(6,0 mL). Purificación: CH<2>Ch/MeOH (9:1). Rendimiento: 108 mg, 18%. Sólido blanco. Pf: 150 - 152 °C. 1H RMN (300 MHz, DMSO-da) 58,48 (t,J= 5,6 Hz, 1H), 7,74 (d,J= 3,9 Hz, 1H), 7,72 - 7,68 (m, 2H), 7,52 (d,J= 3,9 Hz, 1H), 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, 1H), 1,18 (tt,J= 5,4, 11,2 Hz, 2H). 13C RMN (75 MHz, DMSO-da) 5160,7, 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]+ calc. para C<25>H<29>N<2>OS 405,1995; encontrado 405,1996. Pureza (HPLC) > 99 %.Reagents: 5-Phenylthiophene-2-carboxylic acid (1.5 mmol, 306 mg), 2-(1-benzylpiperidin-4-yl)ethan-1-amine (1.53 mmol, 334 mg), EDC (3 mmol, 573 mg), Et<3>N (4.5 mmol, 626 pL) and CH<2>Cl<2>(6.0 mL). Purification: CH<2>Ch/MeOH (9:1). Yield: 108 mg, 18%. White solid. Mp: 150 - 152 °C. 1H NMR (300 MHz, DMSO-da) 58.48 (t,J= 5.6 Hz, 1H), 7.74 (d,J= 3.9 Hz, 1H), 7.72 - 7.68 (m, 2H), 7.52 (d,J= 3.9 Hz, 1H), 7.49 - 7.41 (m, 2H), 7.40 -7.2 1 (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, 1H), 1.18 (tt,J= 5.4, 11.2 Hz, 2H). 13C NMR (75 MHz, DMSO-da) 5160.7, 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, ,1 (2C), 36.8, 35.9, 32.8, 31.7 (2C). HRMS (ESI) m/z: [M H]+ calc. for C<25>H<29>N<2>OS 405,1995; found 405,1996. Purity (HPLC) > 99%.
N-(2-(1-Bencilpiperidin-4-il)etil)-5-bromotiofen-2-carboxamida (3)N-(2-(1-Benzylpiperidin-4-yl)ethyl)-5-bromothiophen-2-carboxamide (3)
Ácido 5-bromo-2-tiofencarboxílico (2 mmol, 414 mg), 2-(1-bencilpiperidin-4-il)etan-1-amina (2,04 mmol, 445 mg), EDC (2 mmol, 310 mg), Et<3>N (4 mmol, 557 pL), HOBt (2 mmol, 270 mg) y CH<2>Ch (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-Ó6) 5 8.51 (t,J= 5,6 Hz, 1H), 7,56 (d,J= 4,0 Hz, 1H), 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, 1H), 1,22 - 1,08 (m, 2H). 13C NMR (75 MHz, DMSO-d6) 5 159,8, 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-thiophenecarboxylic acid (2 mmol, 414 mg), 2-(1-benzylpiperidin-4-yl)ethan-1-amine (2.04 mmol, 445 mg), EDC (2 mmol, 310 mg), Et<3>N (4 mmol, 557 pL), HOBt (2 mmol, 270 mg), and CH<2>Ch (6.0 mL). The product was used in the next step without further purification. Yield: 643 mg, 79%. White solid. 1H NMR (300 MHz, DMSO-Ó6) 5 8.51 (t,J= 5.6 Hz, 1H), 7.56 (d,J= 4.0 Hz, 1H), 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, 1H), 1.22 - 1.08 (m, 2H). 13C NMR (75 MHz, DMSO-d6) 5 159.8, 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-clorofenil)tiofen-2-carboxamida (4)5-Bromo-N-(3-chlorophenyl)thiophen-2-carboxamide (4)
Ácido 5-bromo-2-tiofencarboxílico (1,5 mmol, 310 mg), 3-cloroanilina (1,53 mmol, 194 mg), EDC (1,5 mmol, 233 mg), Et<3>N (3 mmol, 417 ^L), HOBt (1,5 mmol, 203 mg) y CH<2>CI<2>(6,0 mL). Purificación: Hexano/AcOEt 9:1. Rendimiento: 190 mg, 40%. Sólido blanco. 1H NMR (300 MHz, DMSO-cfe) 510,41 (s, 1H), 7,91 - 7,82 (m, 2H), 7,63 (ddd,J= 8,2, 2,1, 1,0 Hz, 1H), 7,43 - 7,35 (m, 2H), 7,17 (ddd,J= 8,0, 2,1, 1,0 Hz, 1H). 13C NMR (75 MHz, DMSO-cfe) 5 158,9, 141,3, 139,9, 133,0, 131,8, 130,4, 130,3, 123,6, 119,7, 118,6, 118,3.5-Bromo-2-thiophenecarboxylic acid (1.5 mmol, 310 mg), 3-chloroaniline (1.53 mmol, 194 mg), EDC (1.5 mmol, 233 mg), Et<3>N (3 mmol, 417 ^L), HOBt (1.5 mmol, 203 mg) and CH<2>Cl<2>(6.0 mL). Purification: Hexane/AcOEt 9:1. Yield: 190 mg, 40%. White solid. 1H NMR (300 MHz, DMSO-cfe) 510.41 (s, 1H), 7.91 - 7.82 (m, 2H), 7.63 (ddd,J= 8.2, 2.1, 1.0 Hz, 1H), 7.43 - 7.35 (m, 2H), 7.17 (ddd,J= 8.0, 2.1, 0Hz, 1H). 13C NMR (75 MHz, DMSO-cfe) 5 158.9, 141.3, 139.9, 133.0, 131.8, 130.4, 130.3, 123.6, 119.7, 118.6, 118.3.
5-Bromo-N-(4-clorofenil)tiofen-2-carboxamida (5)5-Bromo-N-(4-chlorophenyl)thiophen-2-carboxamide (5)
Ácido 5-bromo-2-tiofencarboxílico (3 mmol, 621 mg), 4-cloroanilina (3,06 mmol, 398 mg), EDC (3 mmol, 465 mg), Et<3>N (6 mmol, 835 ^L), HOBt (3 mmol, 405 mg) y CH<2>Cl<2>(10 mL). Purificación: Hexano/AcOEt 7:3. Rendimiento: 368 mg, 39%. Sólido blanco. 1H NMR (300 MHz, DMSO-cfe) 510,39 (s, 1H), 7,85 (d,J= 4,1 Hz, 1H), 7,77 - 7,70 (m, 2H), 7,46 - 7,39 (m, 2H), 7,38 (d,J= 4,1 Hz, 1H). 13C NMR (75 MHz, CDCh) 5158,8, 141,4, 137,4, 131,8, 130,1, 128,7 (2C), 127,6, 121,9 (2C), 118,1.5-Bromo-2-thiophenecarboxylic acid (3 mmol, 621 mg), 4-chloroaniline (3.06 mmol, 398 mg), EDC (3 mmol, 465 mg), Et<3>N (6 mmol, 835 ^L), HOBt (3 mmol, 405 mg) and CH<2>Cl<2>(10 mL). Purification: Hexane/AcOEt 7:3. Yield: 368 mg, 39%. White solid. 1H NMR (300 MHz, DMSO-cfe) 510.39 (s, 1H), 7.85 (d,J= 4.1 Hz, 1H), 7.77 - 7.70 (m, 2H), 7.46 - 7.39 (m, 2H), 7.38 (d,J= 4.1 Hz, 1H). 13C NMR (75 MHz, CDCh) 5158.8, 141.4, 137.4, 131.8, 130.1, 128.7 (2C), 127.6, 121.9 (2C), 118.1.
5-Bromo-N-(2-feniletil)tiofen-2-carboxamida (6)5-Bromo-N-(2-phenylethyl)thiophen-2-carboxamide (6)
Ácido 5-bromo-2-tiofencarboxílico (3 mmol, 621 mg), feniletilamina (3,06 mmol, 383 ^L), EDC (3 mmol, 465 mg), Et<3>N (6 mmol, 835 ^L), HOBt (3 mmol, 405 mg) y CH<2>Cl<2>(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-cfe) 58,68 (t,J= 5.7 Hz, 1H), 7,55 (d,J= 4,0 Hz, 1H), 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-cfe) 5159,9, 141,9, 139,3, 131,5, 128.7 (2C), 128,5, 128,4 (2C), 126,2, 116,5, 40,8, 35,1.5-Bromo-2-thiophenecarboxylic acid (3 mmol, 621 mg), phenylethylamine (3.06 mmol, 383 ^L), EDC (3 mmol, 465 mg), Et<3>N (6 mmol, 835 ^L), HOBt (3 mmol, 405 mg) and CH<2>Cl<2>(10 mL). The product was used in the next step without any further purification. Yield: 253 mg, 56%. White solid. 1H NMR (300 MHz, DMSO-cfe) 58.68 (t,J= 5.7 Hz, 1H), 7.55 (d,J= 4.0 Hz, 1H), 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-cfe) 5159.9, 141.9, 139.3, 131.5, 128.7 (2C), 128.5, 128.4 (2C), 126.2, 116.5, 40.8, 35.1.
5-Bromo-N-(4-morfolinfenil)tiofen-2-carboxamida (7)5-Bromo-N-(4-morpholymphenyl)thiophen-2-carboxamide (7)
Ácido 5-bromo-2-tiofencarboxílico (3 mmol, 621 mg), anilina (3,06 mmol, 561 mg), EDC (3 mmol, 465 mg), Et<3>N (6 mmol, 835 ^L), HOBt (3 mmol, 405 mg) y CH<2>Ch (10 mL). El producto se usó sin ninguna purificación. Rendimiento: 808 mg, 73%. Sólido blanco.5-Bromo-2-thiophenecarboxylic acid (3 mmol, 621 mg), aniline (3.06 mmol, 561 mg), EDC (3 mmol, 465 mg), Et<3>N (6 mmol, 835 ^L), HOBt (3 mmol, 405 mg), and CH<2>Ch (10 mL). The product was used without further purification. Yield: 808 mg, 73%. White solid.
5-Bromo-N-feniltiofen-2-carboxamida (8)5-Bromo-N-phenylthiophen-2-carboxamide (8)
Ácido 5-bromo-2-tiofencarboxílico (3 mmol, 621 mg), anilina (3,06 mmol, 279 ^L), EDC (3 mmol, 465 mg), Et<3>N (6 mmol, 835 ^L), HOBt (3 mmol, 405 mg) y CH<2>Ch (10 mL). Purificación: Hexano/AcOEt 9:1. Rendimiento: 450 mg, 53 %. Sólido blanco. 1H NMR (300 MHz, DMSO-cfe) 510,28 (s, 1H), 7,87 (d,J= 4,1 Hz, 1H), 7,75 - 7,64 (m, 2H), 7,43 - 7,27 (m, 3H), 7,19 - 7,03 (m, 1H). 13C NMR (75 MHz, DMSO) 5 159,2, 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 ^L), EDC (3 mmol, 465 mg), Et<3>N (6 mmol, 835 ^L) , HOBt (3 mmol, 405 mg) and CH<2>Ch (10 mL). Purification: Hexane/AcOEt 9:1. Yield: 450 mg, 53%. White solid. 1H NMR (300 MHz, DMSO-cfe) 510.28 (s, 1H), 7.87 (d,J= 4.1 Hz, 1H), 7.75 - 7.64 (m, 2H), 7, 43 - 7.27 (m, 3H), 7.19 - 7.03 (m, 1H). 13C NMR (75 MHz, DMSO) 5 159.2, 142.3, 138.8, 132.1, 130.3, 129.1, 124.4, 120.8, 118.2.
Síntesis de 5-(h¡drox¡fen¡l)-t¡ofen-2-carboxam¡das (9-20)Synthesis of 5-(hydroxyphenyl)-thiophen-2-carboxamides (9-20)
Esquema 2.Reactivos y condiciones: ii) Pd(PPh<3>)<4>, Na<2>CO<3>, tolueno/H<2>O/EtOH (2:1,5:1), 120 °C (MW).Scheme 2. Reagents and conditions: ii) Pd(PPh<3>)<4>, Na<2>CO<3>, 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), Na<2>CO<3>(2,2 equiv.) y paladio (0) tetrakis(trifenilfosfina) (0,05 equiv.), desgasificada previamente con argón, se añadió una mezcla de tolueno, H<2>O 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 enorto, metaypara.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 120 °C. Finalizado este tiempo, el crudo de reacción se diluyó con una disolución 1:1 de AcOEt/H<2>O y se extrajo con AcOEt. La fase orgánica se lavó con una disolución 1:1 de disolución saturada de NaCl y H<2>O, y finalmente se secó sobre Na<2>SO<4>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: To a mixture of the corresponding 5-bromothiophene-2-carboxamide (1 equiv. obtained by general procedure A), Na<2>CO<3>(2.2 equiv.) and palladium(0) tetrakis(triphenylphosphine) (0.05 equiv.), previously degassed with argon, a mixture of toluene, H<2>O 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 enortho, metaypara substituted derivatives. The reaction mixture was bubbled with argon for a further 15 minutes for complete degassing and then heated under microwave (MW) irradiation for 20 minutes at 120 °C. After this time, the crude reaction mixture 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 and H<2>O, and finally dried over anhydrous Na<2>SO<4>. The desiccant was then filtered and the volatiles were evaporated to dryness in vacuo. The resulting residue was purified by column chromatography using solvent mixtures as eluent, as indicated in each case, to obtain the desired products.
M-(2-(1-Benc¡lp¡per¡d¡n-4-¡l)et¡l)-5-(2-h¡drox¡fen¡l)t¡ofen-2-carboxam¡da (9)Reactivos: N-(2-(1-bencilpiperidin-4-il)etil)-5-bromotiofen-2-carboxamida (3) (1,47 mmol, 601 mg), Na<2>CO<3>(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), H<2>O (3 mL) y EtOH (2 mL). Purificación: CH<2>Ch/MeOH (95:5). Rendimiento: 116 mg, 20%. Sólido blanco. Pf: 185,5 - 187,5 °C. 1H RMN (300 MHz, DMSO-cfe) 510,42 (s, 1H), 8,37 (t,J= 5,6 Hz, 1H), 7,73 - 7,64 (m, 2H), 7,58 (d,J= 4,0 Hz, 1H), 7,35 - 7,20 (m, 5H), 7,16 (td,J= 7,7, 7,2, 1,6 Hz, 1H), 6,97 (dd,J= 8,2, 1,2 Hz, 1H), 6,87 (td,J =7,5, 1,2 Hz, 1H), 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, 1H), 1,23 - 1,08 (m, 2H). 13C RMN (75 MHz, DMSO-cfe) 5 161,3, 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]+ calc. para C<25>H<29>N<2>O<2>S 421,1944; encontrado 421,1946. Pureza (HPLC) > 99%.M-(2-(1-Benc¡lp¡perídin-4-¡l)et¡l)-5-(2-hydroxy¡phen¡l)thiophen-2-carboxamide (9)Reagents: N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-bromothiophen-2-carboxamide (3) (1.47 mmol, 601 mg), Na<2>CO<3> (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), H<2>O (3 mL) and EtOH (2 mL). Purification: CH<2>Ch/MeOH (95:5). Yield: 116 mg, 20%. White solid. Mp: 185.5 - 187.5 °C. 1H NMR (300 MHz, DMSO-cfe) 510.42 (s, 1H), 8.37 (t,J= 5.6 Hz, 1H), 7.73 - 7.64 (m, 2H), 7, 58 (d,J= 4.0 Hz, 1H), 7.35 - 7.20 (m, 5H), 7.16 (td,J= 7.7, 7.2, 1.6 Hz, 1H) , 6.97 (dd,J= 8.2, 1.2 Hz, 1H), 6.87 (td,J =7.5, 1.2 Hz, 1H), 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, 1H), 1 .23 - 1.08 (m, 2H). 13C NMR (75 MHz, DMSO-cfe) 5 161.3, 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]+ calc. for C<25>H<29>N<2>O<2>S 421,1944; found 421,1946. Purity (HPLC) > 99%.
M-(2-(1-Bencilpiperidin-4-il)etil)-5-(3-hidroxifenil)tiofen-2-carboxamida (10) Reactivos: N-(2-(1-bencilpiperidin-4-il)etil)-5-bromotiofen-2-carboxamida (3) (2,21 mmol, 900 mg), Na<2>CO<3>(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), H<2>O (3 mL) y EtOH (2 mL). Purificación: CH<2>Ch/MeOH (95:5). Rendimiento: 754 mg, 80%. Sólido blanco. Pf: 159 - 161 °C. 1H RMN (300 MHz, DMSO-da) 59,68 (s, 1H), 8,46 (t,J= 5,6 Hz, 1H), 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, 1H), 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).13C RMN (75 MHz, DMSO-da) 5 160,8, 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-(2-(1-Benzylpiperidin-4-yl)ethyl)-5-(3-hydroxyphenyl)thiophene-2-carboxamide (10) Reagents: N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-bromothiophene-2-carboxamide (3) (2.21 mmol, 900 mg), Na<2>CO<3>(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 solution of toluene (4 mL), H<2>O (3 mL) and EtOH (2 mL). Purification: CH<2>Ch/MeOH (95:5). Yield: 754 mg, 80%. White solid. Mp: 159 - 161 °C. 1H NMR (300 MHz, DMSO-da) 59.68 (s, 1H), 8.46 (t,J= 5.6 Hz, 1H), 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 , 1H), 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).13C NMR (75 MHz, DMSO-da) 5 160.8, 157.9, 147.3, 138.9, 134.3, 130.3, 128.8, 128.1 (5 C), 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 C<25>H<29>N<2>O<2>S 421,1944; encontrado 421,1943. Pureza (HPLC): 96%.[M H]+ calc. for C<25>H<29>N<2>O<2>S 421,1944; found 421,1943. Purity (HPLC): 96%.
M-(2-(1-Bencilpiperidin-4-il)etil)-5-(4-hidroxifenil)tiofen-2-carboxamida (11) Reactivos: N-(2-(1-bencilpiperidin-4-il)etil)-5-bromotiofen-2-carboxamida (3) (2,21 mmol, 900 mg), Na<2>CO<3>(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), H<2>O (3 mL) y EtOH (2 mL). Purificación: CH<2>Ch/MeOH (95:5). Rendimiento: 550 mg, 60%. Sólido blanco. Pf: 164 - 166 °C. 1H RMN (300 MHz, DMSO-da) 59,77 (s, 1H), 8,37 (t,J= 5,4 Hz, 1H), 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, 1H), 1,23 - 1,08 (m, 2H).13C RMN (75 MHz, DMSO-^) 5160,9, 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 C<25>H<29>N<2>O<2>S 421,1944; encontrado 421,1948. Pureza (HPLC): 98%.M-(2-(1-Benzylpiperidin-4-yl)ethyl)-5-(4-hydroxyphenyl)thiophene-2-carboxamide (11) Reagents: N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-bromothiophene-2-carboxamide (3) (2.21 mmol, 900 mg), Na<2>CO<3>(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: CH<2>Ch/MeOH (95:5). Yield: 550 mg, 60%. White solid. Mp: 164 - 166 °C. 1H NMR (300 MHz, DMSO-da) 59.77 (s, 1H), 8.37 (t,J= 5.4 Hz, 1H), 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, 1H), 1.23 - 1.08 (m, 2H).13C NMR (75 MHz, DMSO-^) 5160.9, 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, 5 3.2 (2C), 36.8, 35.9, 32.9, 31.8 (2C). HRMS (ESI) m/z: [M H]+ calc. for C<25>H<29>N<2>O<2>S 421,1944; found 421,1948. Purity (HPLC): 98%.
N-(3-Clorofenil)-5-(2-hidroxifenil)tiofen-2-carboxamida (12)N-(3-Chlorophenyl)-5-(2-hydroxyphenyl)thiophen-2-carboxamide (12)
Reactivos: 5-bromo-N-(3-clorofenil)-2-tiofencarboxamida (4) (0,84 mmol, 265 mg), Na<2>CO<3>(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 ^L) y una solución de tolueno (2 mL), H<2>O (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-de)510,56 (s, 1H), 10,31 (s, 1H), 8,00 (d,J= 4,1 Hz, 1H), 7,94 (t,J= 2,0 Hz, 1H,), 7,77 (dd,J= 7,9, 1,7 Hz, 1H), 7,73 - 7,67 (m, 2H), 7,39 (t,J= 8,1 Hz, 1H), 7,24 - 7,13 (m, 2H), 7,01 (dd,J= 8,2, 1,2 Hz, 1H), 6,91 (ddd,J= 7,8, 7,2, 1,2 Hz, 1H). 13C RMN (75 MHz, DMSO-de) 5 160,5, 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 C<17>H<13>ClNO<2>S 330,0350; encontrado 330,0348. Pureza (HPLC): 96,0%.Reagents: 5-bromo-N-(3-chlorophenyl)-2-thiophenecarboxamide (4) (0.84 mmol, 265 mg), Na<2>CO<3>(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 ^L) and a solution of toluene (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. 1H NMR (300 MHz, DMSO-de)510.56 (s, 1H), 10.31 (s, 1H), 8.00 (d,J= 4.1 Hz, 1H), 7.94 (t,J= 2.0 Hz, 1H,), 7.77 (dd,J= 7.9, 1.7 Hz, 1H), 7.73 - 7.67 (m, 2H), 7.39 (t,J= 8.1 Hz, 1H), 7.24 - 7.13 (m, 2H), 7.01 (dd,J= 8.2, 1.2 Hz, 1H), 6.91 (ddd,J= 7.8, 7.2, 1.2 Hz, 1H). 13C NMR (75 MHz, DMSO-de) 5 160.5, 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, 8.5, 116.4. HRMS (ESI) m/z: [M H]+ calc. for C<17>H<13>ClNO<2>S 330.0350; found 330.0348. Purity (HPLC): 96.0%.
N-(3-Clorofenil)-5-(3-hidroxifenil)tiofen-2-carboxamida (13)N-(3-Chlorophenyl)-5-(3-hydroxyphenyl)thiophen-2-carboxamide (13)
Reactivos: 5-bromo-N-(3-clorofenil)tiofen-2-carboxamida (4) (0,32 mmol, 101 mg), Na<2>CO<3>(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), H<2>O (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-da) 510,37 (s, 1H), 9,70 (s, 1H), 8,01 (d,J= 3,9 Hz, 1H), 7,92 (t,J= 2,0 Hz, 1H), 7,72 -7,63 (m, 1H), 7,55 (d,J= 4,0 Hz, 1H), 7,39 (t,J= 8,1 Hz, 1H), 7,26 (t, 1H), 7,21 - 7,14 (m, 2H), 7,13 - 7,08 (m, 1H), 6,83 - 6,77 (m, 1H). 13C RMN (75 MHz, DMSO) 5 159,9, 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 C<17>H<13>CINO<2>S 330,0350; encontrado 330,0347. Pureza (HPLC) > 99%.Reagents: 5-bromo-N-(3-chlorophenyl)thiophene-2-carboxamide (4) (0.32 mmol, 101 mg), Na<2>CO<3>(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 solution of toluene (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. 1H NMR (300 MHz, DMSO-da) 510.37 (s, 1H), 9.70 (s, 1H), 8.01 (d,J= 3.9 Hz, 1H), 7.92 (t,J= 2.0 Hz, 1H), 7.72 -7.63 (m, 1H), 7.55 (d,J= 4.0 Hz, 1H), 39 (t,J= 8.1 Hz, 1H), 7.26 (t, 1H), 7.21 - 7.14 (m, 2H), 7.13 - 7.08 (m, 1H), 6.83 - 6.77 (m, 1H). 13C NMR (75 MHz, DMSO) 5 159.9, 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 C<17>H<13>CINO<2>S 330.0350; found 330.0347. Purity (HPLC) > 99%.
N-(3-Clorofenil)-5-(4-hidroxifenil)tiofen-2-carboxamida (14)N-(3-Chlorophenyl)-5-(4-hydroxyphenyl)thiophen-2-carboxamide (14)
Reactivos: 5-bromo-N-(3-clorofenil)tiofen-2-carboxamida (4) (1,27 mmol, 400 mg), Na<2>CO<3>(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), H<2>O (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) 510,31 (s, 1H), 9,84 (s, 1H), 7,97 (d,J= 4,0 Hz, 1H), 7,91 (t,J= 2,0 Hz, 1H), 7,67 (ddd,J= 8,3, 2,1, 1,0 Hz, 1H), 7,61 - 7,54 (m, 2H), 7,43 (d,J= 4,0 Hz, 1H), 7,38 (t,J= 8,1 Hz, 1H), 7,15 (ddd,J= 8,0, 2,1, 0,9 Hz, 1H), 6,88 - 6,81 (m, 2H).13C RMN (75 MHz, DMSO-de) 5 159,9, 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 C<17>H<13>CINO<2>S 330,0350; encontrado 330,0346. Pureza (HPLC) > 99%.Reagents: 5-bromo-N-(3-chlorophenyl)thiophene-2-carboxamide (4) (1.27 mmol, 400 mg), Na<2>CO<3>(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 solution of toluene (3.1 mL), H<2>O (2.33 mL) and EtOH (1.57 mL). Purification: Hexane/AcOEt 9:1. Yield: 73 mg, 18%. Yellow solid. Mp: decomposed. 1H NMR (300 MHz, DMSO-d6) 510.31 (s, 1H), 9.84 (s, 1H), 7.97 (d,J= 4.0 Hz, 1H), 7.91 (t,J= 2.0 Hz, 1H), 7.67 (ddd,J= 8.3, 2.1, 1.0 Hz, 1H), 7.61 - 7 13C NMR (75 MHz, DMSO-de) 5 159.9, 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 C<17>H<13>CINO<2>S 330.0350; found 330.0346. Purity (HPLC) > 99%.
N-(4-Clorofenil)-5-(2-hidroxifenil)tiofen-2-carboxamida (15)N-(4-Chlorophenyl)-5-(2-hydroxyphenyl)thiophen-2-carboxamide (15)
Reactivos: 5-bromo-N-(4-clorofenil)tiofen-2-carboxamida (5) (1,16 mmol, 368 mg), Na<2>CO<3>(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 solución de tolueno (2 mL), H<2>O (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-de) 510,56 (s, 1H), 10,29 (s, 1H), 7,98 (d,J= 4,1 Hz, 1H), 7,82 - 7,73 (m, 3H), 7,70 (d,J= 4,1 Hz, 1H), 7,46 - 7,37 (m, 2H), 7,20 (ddd,J= 8,3, 7,3, 1,6 Hz, 1H), 6,99 (dd,J= 8,2, 1,2 Hz, 1H), 6,90 (ddd,J= 8,3, 7,2, 1,2 Hz, 1H). 13C RMN (75 MHz, DMSO-de) 5 160,3, 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 C<1t>H<13>CINO<2>S 330,0350; encontrado 330,0335. Pureza (HPLC): 98,3%.Reagents: 5-bromo-N-(4-chlorophenyl)thiophene-2-carboxamide (5) (1.16 mmol, 368 mg), Na<2>CO<3>(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 solution of toluene (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. 1H NMR (300 MHz, DMSO-de) 510.56 (s, 1H), 10.29 (s, 1H), 7.98 (d,J= 4.1 Hz, 1H), 7.82 - 7.73 (m, 3H), 7.70 (d,J= 4.1 Hz, 1H), 7.46 - 7.37 (m, 2H), 7. 20 (ddd,J= 8.3, 7.3, 1.6 Hz, 1H), 6.99 (dd,J= 8.2, 1.2 Hz, 1H), 6.90 (ddd,J= 8.3, 7.2, 1.2 Hz, 1H). 13C NMR (75 MHz, DMSO-de) 5 160.3, 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, , 116.4. HRMS (ESI) m/z: [M H]+ calc. for C<1t>H<13>CINO<2>S 330.0350; found 330.0335. Purity (HPLC): 98.3%.
N-(4-Clorofenil)-5-(3-hidroxifenil)tiofen-2-carboxamida (16)N-(4-Chlorophenyl)-5-(3-hydroxyphenyl)thiophen-2-carboxamide (16)
Reactivos: 5-bromo-N-(4-clorofenil)tiofen-2-carboxamida (5) (1,28 mmol, 404 mg), Na<2>CO<3>(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), H<2>O (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-da) 5 10,35 (s, 1H), 9,71 (s, 1H), 8,00 (d,J= 4,0 Hz, 1H), 7,83 - 7,73 (m, 2H), 7,55 (d,J= 3,9 Hz, 1H), 7,48 - 7,38 (m, 2H), 7,26 (t,J= 7,8 Hz), 7,17 (dt,J= 7,7, 1,3 Hz, 1H), 7,10 (t,J= 2,1 Hz, 1H), 6,80 (ddd,J= 8,0, 2,4, 1,1 Hz, 1H). 13C RMN (75 MHz, DMSO) 5 159,7, 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 C<1t>H<13>CINO<2>S 330,0350; encontrado 330,0355. Pureza (HPLC) > 99%.Reagents: 5-bromo-N-(4-chlorophenyl)thiophene-2-carboxamide (5) (1.28 mmol, 404 mg), Na<2>CO<3>(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 solution of toluene (3.1 mL), H<2>O (2.33 mL) and EtOH (1.57 mL). Purification: Hexane/AcOEt 9:1. Yield: 180 mg, 43%. Yellow solid. Mp: decomposed. 1H NMR (300 MHz, DMSO-da) 5 10.35 (s, 1H), 9.71 (s, 1H), 8.00 (d,J= 4.0 Hz, 1H), 7.83 - 7.73 (m, 2H), 7.55 (d,J= 3.9 Hz, 1H), 7.48 - 7.38 (m, 2H), 7. 26 (t,J= 7.8 Hz), 7.17 (dt,J= 7.7, 1.3 Hz, 1H), 7.10 (t,J= 2.1 Hz, 1H), 6.80 (ddd,J= 8.0, 2.4, 1.1 Hz, 1H). 13C NMR (75 MHz, DMSO) 5 159.7, 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 C<1t>H<13>CINO<2>S 330.0350; found 330.0355. Purity (HPLC) > 99%.
N-(4-Clorofenil)-5-(4-hidroxifenil)tiofen-2-carboxamida (17)N-(4-Chlorophenyl)-5-(4-hydroxyphenyl)thiophen-2-carboxamide (17)
Reactivos: 5-bromo-N-(4-clorofenil)tiofen-2-carboxamida (5) (1,58 mmol, 500 mg), Na<2>CO<3>(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), H<2>O (3 mL) y EtOH (2 mL). Purificación: Hexano/AcOEt 9:1.Reagents: 5-bromo-N-(4-chlorophenyl)thiophene-2-carboxamide (5) (1.58 mmol, 500 mg), Na<2>CO<3>(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 solution of toluene (4 mL), H<2>O (3 mL) and EtOH (2 mL). Purification: Hexane/AcOEt 9:1.
Rendimiento: 208 mg, 40%. Sólido amarillo. Pf: 291 - 293 °C. 1H RMN (300 MHz, DMSO-da) 5 10,28 (s, 1H), 9,83 (s, 1H), 7,96 (d,J= 4,0 Hz, 1H), 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) 5 159,9, 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 C<17>H<13>CINO<2>S 330,0350; encontrado 330,0351. Pureza (HPLC) > 99%.Yield: 208 mg, 40%. Yellow solid. Mp: 291 - 293 °C. 1H NMR (300 MHz, DMSO-da) 5 10.28 (s, 1H), 9.83 (s, 1H), 7.96 (d,J= 4.0 Hz, 1H), 7.81 - 7.72 (m, 2H), 7.62 - 7.52 (m, 2H), 7.46 - 7.37 (m, 3H), - 6.78 (m, 2H).13C NMR (75 MHz, DMSO) 5 159.9, 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 C<17>H<13>CINO<2>S 330.0350; found 330.0351. Purity (HPLC) > 99%.
N-Fenetil-5-(2-hidroxifenil)tiofen-2-carboxamida (18)N-Phenethyl-5-(2-hydroxyphenyl)thiophen-2-carboxamide (18)
Reactivos: 5-bromo-N-(2-feniletil)-2-tiofencarboxamida (6) (2,23 mmol, 692 mg), Na<2>CO<3>(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 ^L) y una solución de tolueno (4 mL), H<2>O (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-da) 510,42 (s, 1H), 8,53 (t,J= 5,7 Hz, 1H), 7,72 - 7,64 (m, 2H), 7,58 (d,J= 4,0 Hz, 1H), 7,34 - 7,13 (m, 6H), 6,97 (dd,J= 8,2, 1,2 Hz, 1H), 6,87 (td,J= 7,5, 1,3 Hz, 1H), 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-da) 5 161,4, 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]+ calc. para C<19>H<18>NO<2>S 324,1053; encontrado 324,1043. Pureza (HPLC)> 99%.Reagents: 5-bromo-N-(2-phenylethyl)-2-thiophenecarboxamide (6) (2.23 mmol, 692 mg), Na<2>CO<3>(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 ^L), and a solution of toluene (4 mL), H<2>O (3 mL), and EtOH (2 mL). The product was used without further purification. Yield: 263 mg, 45%. White solid. Mp: 200 - 202 °C. 1H NMR (300 MHz, DMSO-da) 510.42 (s, 1H), 8.53 (t,J= 5.7 Hz, 1H), 7.72 - 7.64 (m, 2H), 7.58 (d,J= 4.0 Hz, 1H), 7.34 - 7.13 (m, 6H), 6.97 (dd,J= 8.2, 1.2 Hz, 1H), 6.87 (td,J= 7.5, 1.3 Hz, 1H), 3.46 (dt,J= 8.0, 6.1 Hz, 2H), 2.84 (dd,J= 8.3, 6.6 Hz, 2H). 13C NMR (75 MHz, DMSO-da) 5 161.4, 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, 40.7, 35.2. HRMS (ESI) m/z: [M H]+ calc. for C<19>H<18>NO<2>S 324.1053; found 324.1043. Purity (HPLC) > 99%.
N-Fenetil-5-(3-hidroxifenil)tiofen-2-carboxamida (19)N-Phenethyl-5-(3-hydroxyphenyl)thiophen-2-carboxamide (19)
Reactivos: 5-bromo-N-(2-feniletil)-2-tiofencarboxamida (6) (1,13 mmol, 350 mg), Na<2>CO<3>(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), H<2>O (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-da) 59,67 (s, 1H), 8,62 (t,J= 5,6 Hz, 1H), 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, 1H), 7,05 (t,J= 2,0 Hz, 1H), 6,77 (ddd,J= 8,0, 2,4, 1,0 Hz, 1H), 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) 5160,9, 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:Reagents: 5-bromo-N-(2-phenylethyl)-2-thiophenecarboxamide (6) (1.13 mmol, 350 mg), Na<2>CO<3>(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 solution of toluene (2 mL), H<2>O (1.5 mL) and EtOH (1 mL). Purification: Hexane/AcOEt 1:1. Yield: 70 mg, 19%. White solid. Mp: 183 - 185 °C. 1H NMR (300 MHz, DMSO-da) 59.67 (s, 1H), 8.62 (t,J= 5.6 Hz, 1H), 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 13C NMR (75 MHz, DMSO) 5160.9, 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, 4 0.8, 35.2. HRMS (ESI) m/z:
[M H]+ calc. para C<19>H<18>NO<2>S 324,1053; encontrado 324,1049. Pureza (HPLC) > 99%.[M H]+ calc. for C<19>H<18>NO<2>S 324.1053; found 324.1049. Purity (HPLC) > 99%.
5-(2-Hidroxifenil)-N-(4-morfolinofenil)tiofen-2-carboxamida (20)5-(2-Hydroxyphenyl)-N-(4-morpholinophenyl)thiophen-2-carboxamide (20)
Reactivos: 5-bromo-N-(4-morfolinofenil)tiofen-2-carboxamida (7) (2,2 mmol, 809 mg), Na<2>CO<3>(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), H<2>O (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 °C.Reagents: 5-bromo-N-(4-morpholinophenyl)thiophene-2-carboxamide (7) (2.2 mmol, 809 mg), Na<2>CO<3>(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 solution of toluene (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 °C.
1H RMN (300 MHz, DMSO-cfe) 59,99 (s, 1H), 7,92 (d,J= 4,1 Hz, 1H), 7,72 (dd,J= 8,0, 1,6 Hz, 1H), 7,65 (d,J= 4,1 Hz, 1H), 7,63 - 7,56 (m, 2H), 7,13 (td,J= 7,6, 7,1, 1,6 Hz, 1H), 7,01 (dd,J= 8,3, 1,2 Hz, 1H), 6,97 - 6,89 (m, 2H), 6,85 - 6,75 (m, 1H), 3,80 - 3,67 (m, 4H), 3,13 - 3,00 (m, 4H). 13C RMN (75 MHz, DMSO-cfe) 5160,0, 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]+ calc. para C<21>H<20>N<2>O<3>S 381,1267; encontrado 381,1267. Pureza (HPLC): 97,8%.1H NMR (300 MHz, DMSO-cfe) 59.99 (s, 1H), 7.92 (d,J= 4.1 Hz, 1H), 7.72 (dd,J= 8.0, 1.6 Hz, 1H), 7.65 (d,J= 4.1 Hz, 1H), 7.63 - 7.56 (m, 2H), 7.13 (td,J= 7.6, 7.1, 1.6 Hz, 1H), 7.01 (dd,J= 8.3, 1.2 Hz, 1H), 6.97 - 6.89 (m, 2H), 6.85 - 6.75 (m , 1H), 3.80 - 3.67 (m, 4H), 3.13 - 3.00 (m, 4H). 13C NMR (75 MHz, DMSO-cfe) 5160.0, 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]+ calc. for C<21>H<20>N<2>O<3>S 381.1267; found 381.1267. Purity (HPLC): 97.8%.
Síntesis de (piperidin-4-iloxi)fenil)tiofen-2-carboxamidas (21-30)Synthesis of (piperidin-4-yloxy)phenyl)thiophen-2-carboxamides (21-30)
Esquema 3.Reactivos y condiciones. (iii) DIAD, PPh<3>, THF, 0 °C a 25 °C; (iv) TFA, CH<2>CE 25 °C.Scheme 3. Reagents and conditions. (iii) DIAD, PPh<3>, THF, 0 °C to 25 °C; (iv) TFA, CH<2>CE 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 °C, 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 terc-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 CH<2>Ch/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 CH<2>Ch se basificó con NaHCO<3>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: To a solution of triphenylphosphine (1.3 eq) in THF at 0 °C was added DIAD (1.3 eq). The reaction mixture was stirred at room temperature until a white turbidity appeared. After cooling again to 0 °C, a solution of the precursor 5-(2-hydroxyphenyl)-thiophene-2-carboxamide (1 equiv., obtained by general procedure B) and tert-butyl 4-hydroxypiperidine-1-carboxylate (1.3 equiv.) in THF was added. The mixture 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 CH<2>Ch/TFA solution and stirred at room temperature for 3 hours. After deprotection, the solvent was evaporated in vacuo and after dissolving the resulting product in CH<2>Ch, 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 the solvent mixtures indicated in each case as eluent to obtain the desired products.
M-(2-(1-Bencilpiperidin-4-il)etil)-5-(2-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (21)M-(2-(1-Benzylpiperidin-4-yl)ethyl)-5-(2-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide (21)
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 terc-butilo (2,16 mmol, 434 mg), azodicarboxilato de diisopropilo (2,16 mmol, 426 pL), 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 CH<2>Cl<2>(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-da) 58,39 (t,J= 5,7 Hz, 1H), 7,77 (dd,J= 7,9, 1,7 Hz, 1H), 7,68 (d,J= 4,0 Hz, 1H), 7,58 (d,J= 4,0 Hz, 1H), 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, 1H), 1,20 - 1,12 (m, 2H).13C RMN (75 MHz, DMSO-da) 5161,3, 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]+ calc. para C<30>H<38>N<3>O<2>S 504,2679; encontrado 504,2680. Pureza (HPLC): 98,7%.Reagents: N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-(2-hydroxyphenyl)thiophene-2-carboxamide (9) (1.66 mmol, 700 mg), tert-butyl 4-hydroxypiperidine-1-carboxylate (2.16 mmol, 434 mg), diisopropyl azodicarboxylate (2.16 mmol, 426 pL), triphenylphosphine (2.16 mmol, 434 mg) and 15 mL of THF. 150 mg of tert-butyl 4-(5-((2-(1-benzylpiperidin-4-yl)ethyl)carbamoyl)thiophen-2-yl)phenoxy)piperidine-1-carboxylate were obtained, which were treated with a solution of CH<2>Cl<2>(3 mL) and trifluoroacetic acid (2 mL). Purification: AcOEt/MeOH 8:2. Yield: 30 mg, 24%. White solid. Mp: 118 - 120 °C. 1H NMR (300 MHz, DMSO-da) 58.39 (t,J= 5.7 Hz, 1H), 7.77 (dd,J= 7.9, 1.7 Hz, 1H), 7.68 (d,J= 4.0 Hz, 1H), 7.58 (d,J= 4.0 Hz, 1H), 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, 1H), 1.20 - 1.12 (m, 2H).13C NMR (75 MHz, DMSO-da) 5161.3, 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]+ calc. for C<30>H<38>N<3>O<2>S 504.2679; found 504.2680. Purity (HPLC): 98.7%.
N-(3-Clorofenil)-5-(2-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (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-clorofenil)carbamoil)tiofen-2-il)fenoxi)piperidin-1-carboxilato de terc-butilo que se trataron con una solución de CH<2>Cl<2>(3 mL) y ácido trifluoroacético (2 mL). Purificación: CH<2>Cl<2>/MeOH 9:1. Rendimiento: 53 mg, 39%. Sólido blanco. Pf: 195 - 197 °C. 1H RMN (500 MHz, DMSO-da) 510,42 (s, 1H), 8,06 (d,J= 4,1 Hz, 1H), 7,96 (t,J= 2,1 Hz, 1H), 7,86 (dd,J= 7,9, 1,7 Hz, 1H), 7,73 - 7,68 (m, 2H), 7,38 (t,J= 8,1 Hz, 1H), 7,33 (ddd,J= 8,7, 7,2, 1,7 Hz, 1H), 7,23 (d,J= 8,6 Hz, 1H), 7,15 (ddd,J= 8,0, 2,1,0,9 Hz, 1H), 7,05 - 7,01 (m, 1H), 4,72 - 4,66 (m, 1H), 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-Ó6) 5160,5, 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]+ calc. para C<22>H<22>CIN<2>O<2>S 413,1085; encontrado 413,1083. Pureza (HPLC) > 99%.N-(3-Chlorophenyl)-5-(2-(piperidin-4-yloxy)phenyl)thiophene-2-carboxamide (22)Reagents: N-(3-chlorophenyl)-5-(2-hydroxyphenyl)thiophene-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-chlorophenyl)carbamoyl)thiophen-2-yl)phenoxy)piperidine-1-carboxylate were obtained, which were treated with a solution of CH<2>Cl<2>(3 mL) and trifluoroacetic acid (2 mL). Purification: CH<2>Cl<2>/MeOH 9:1. Yield: 53 mg, 39%. White solid. Mp: 195 - 197 °C. 1H NMR (500 MHz, DMSO-da) 510.42 (s, 1H), 8.06 (d,J= 4.1 Hz, 1H), 7.96 (t,J= 2.1 Hz, 1H), 7.86 (dd,J= 7.9, 1.7 Hz, 1H), 7.73 - 7.68 (m, 2H), 7.38 (t,J = 8.1 Hz, 1H), 7.33 (ddd,J= 8.7, 7.2, 1.7 Hz, 1H), 7.23 (d,J= 8.6 Hz, 1H), 7.15 (ddd,J= 8.0, 2.1.0.9 Hz, 1H), 7.05 - 7.01 (m, 1H), 4.72 - 4.66 (m, 1H), 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 NMR (125 MHz, DMSO-Ó6) 5160.5, 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, 6, 118.5, 114.3, 74.4, 43.4 (2C), 31.9 (2C). HRMS (ESI) m/z: [M H]+ calc. for C<22>H<22>CIN<2>O<2>S 413.1085; found 413.1083. Purity (HPLC) > 99%.
N-(3-Clorofenil)-5-(3-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (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 CH<2>Ch (3 mL) y ácido trifluoroacético (2 mL). Purificación: CH<2>Ch/MeOH 9:1. Rendimiento: 17 mg, 16 %. Sólido amarillo. Pf: 150 - 152 °C. 1H RMN (300 MHz, DMSO-cfe) 5 10,40 (s, 1H), 8,03 (d,J= 4,0 Hz, 1H), 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, 1H).N-(3-Chlorophenyl)-5-(3-(piperidin-4-yloxy)phenyl)thiophene-2-carboxamide (23)Reagents: N-(3-chlorophenyl)-5-(3-hydroxyphenyl)thiophene-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 tert-butyl 4-(5-((3-chlorophenyl)carbamoyl)thiophen-2-yl)phenoxy)piperidine-1-carboxylate were obtained, which were treated with a solution of CH<2>Ch (3 mL) and trifluoroacetic acid (2 mL). Purification: CH<2>Ch/MeOH 9:1. Yield: 17 mg, 16 %. Yellow solid. Mp: 150 - 152 °C. 1H NMR (300 MHz, DMSO-cfe) 5 10.40 (s, 1H), 8.03 (d,J= 4.0 Hz, 1H), 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, 1H).
13C RMN (75 MHz, DMSO-cfe) 5159,9, 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]+ calc. para C<22>H<22>ClN<2>O<2>S 413,1085; encontrado 413,1080. Pureza (HPLC) > 99%.13C NMR (75 MHz, DMSO-cfe) 5159.9, 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]+ calc. for C<22>H<22>ClN<2>O<2>S 413.1085; found 413.1080. Purity (HPLC) > 99%.
N-(3-clorofenil)-5-(4-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (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 CH<2>Ch (4 mL) y ácido trifluoroacético (1 mL). Purificación: CH<2>Ch/MeOH 9:1. Rendimiento: 38 mg, 22%. Sólido blanco. Pf: 224 - 226 °C. 1H RMN (300 MHz, DMSO-cfe) 5 10,37 (s, 1H), 8,02 (d,J= 4,0 Hz, 1H), 7,92 (t,J= 2,0 Hz, 1H), 7,73 - 7,61 (m, 3H), 7,50 (d,J= 3,9 Hz, 1H), 7,39 (t,J= 8,1 Hz, 1H), 7,20 - 7,13 (m, 1H), 7,09 - 6,99 (m, 2H), 4,58 -4,40 (m, 1H), 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) 5 159,9, 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]+ calc. para C<22>H<22>ClN<2>O<2>S 413,1085; encontrado 413,1083. Pureza (HPLC) > 99%.N-(3-chlorophenyl)-5-(4-(piperidin-4-yloxy)phenyl)thiophene-2-carboxamide (24)Reagents: N-(3-chlorophenyl)-5-(4-hydroxyphenyl)thiophene-2-carboxamide (14) (0.88 mmol, 290 mg), tert-butyl 4-hydroxypiperidine-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 solution of CH<2>Ch (4 mL) and trifluoroacetic acid (1 mL). Purification: CH<2>Ch/MeOH 9:1. Yield: 38 mg, 22%. White solid. Mp: 224 - 226 °C. 1H NMR (300 MHz, DMSO-cfe) 5 10.37 (s, 1H), 8.02 (d,J= 4.0 Hz, 1H), 7.92 (t,J= 2.0 Hz, 1H), 7.73 - 7.61 (m, 3H), 7.50 (d,J= 3.9 Hz, 1H), 7.39 (t,J= 8 ,1 Hz, 1H), 7.20 - 7.13 (m, 1H), 7.09 - 6.99 (m, 2H), 4.58 -4.40 (m, 1H), 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 NMR (75 MHz, DMSO) 5 159.9, 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, 4 (2C), 72.9, 43.2 (2C), 31.5 (2C). HRMS (ESI) m/z: [M H]+ calc. for C<22>H<22>ClN<2>O<2>S 413.1085; found 413.1083. Purity (HPLC) > 99%.
N-(4-Clorofenil)-5-(2-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (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 CH<2>Cl<2>(5 mL) y ácido trifluoroacético (3,3 mL). Purificación: la mezcla orgánica se evaporó hasta sequedad a vacío y a continuación se disolvió en CH<2>Ch y lavó con NaOH 1M, salmuera y agua. La fase orgánica se acidificó con HCl 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)5 10,29 (s, 1H) 7,93 (d,J= 4,1 Hz, 1H), 7,82 (dd,J= 7,9, 1,7 Hz, 1H), 7,78 - 7,75 (m, 2H), 7,66 (d,J= 4,2 Hz, 1H), 7,40 - 7,35 (m, 2H), 7,31 (ddd,J= 8,6, 7,2, 1,7 Hz, 1H), 7,21 (d,J= 8,5 Hz, 1H), 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). 13C RMN (75 MHz, DMSO-da) 5160,6, 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]+ calc. para C<22>H<22>ClN<2>O<2>S 413,1085; encontrado 413,1077. Pureza (HPLC): 98,7%.N-(4-Chlorophenyl)-5-(2-(piperidin-4-yloxy)phenyl)thiophene-2-carboxamide (25)Reagents: N-(4-chlorophenyl)-5-(2-hydroxyphenyl)thiophene-2-carboxamide (15) (0.76 mmol, 250 mg), tert-butyl 4-hydroxypiperidine-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 solution of CH<2>Cl<2>(5 mL) and trifluoroacetic acid (3.3 mL). Purification: The organic mixture was evaporated to dryness in vacuo and then dissolved in CH<2>Ch 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 the appearance of a white precipitate. Yield: 30.5 mg, 16%. White solid. Mp: 143 - 145 °C. 1H NMR (300 MHz, DMSO-d6)5 10.29 (s, 1H) 7.93 (d,J= 4.1 Hz, 1H), 7.82 (dd,J= 7.9, 1.7 Hz, 1H), 7.78 - 7.75 (m, 2H), 7.66 (d,J= 4.2 Hz, 1H), 7.40 - 7 .35 (m, 2H), 7.31 (ddd,J= 8.6, 7.2, 1.7 Hz, 1H), 7.21 (d,J= 8.5 Hz, 1H), 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). 13C NMR (75 MHz, DMSO-da) 5160.6, 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, 7 4.7, 43.6 (2C), 32.2 (2C). HRMS (ESI) m/z: [M H]+ calc. for C<22>H<22>ClN<2>O<2>S 413.1085; found 413.1077. Purity (HPLC): 98.7%.
N-(4-Clorofenil)-5-(3-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (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 CH<2>Ch (8 mL) y ácido trifluoroacético (2 mL). Purificación: CH<2>Ch/MeOH 9:1. Rendimiento: 137 mg, 34%. Sólido blanco. Pf: 169 - 171 °C. 1H RMN (300 MHz, DMSO-d6) 510,36 (s, 1H), 8,02 (d,J= 4,0 Hz, 1H), 7,82 - 7,74 (m, 2H), 7,65 (d,J= 3,9 Hz, 1H), 7,47 - 7,39 (m, 2H), 7,35 (t,J= 8,1 Hz, 1H), 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).13C RMN (75 MHz, DMSO-d6) 5 159,7, 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]+ calc. para C<22>H<22>ClN<2>O<2>S 413,1085; encontrado 413,1088. Pureza (HPLC) > 99%.N-(4-Chlorophenyl)-5-(3-(piperidin-4-yloxy)phenyl)thiophene-2-carboxamide (26)Reagents: N-(4-chlorophenyl)-5-(3-hydroxyphenyl)thiophene-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 CH<2>Ch (8 mL) and trifluoroacetic acid (2 mL). Purification: CH<2>Ch/MeOH 9:1. Yield: 137 mg, 34%. White solid. Mp: 169 - 171 °C. 1H NMR (300 MHz, DMSO-d6) 510.36 (s, 1H), 8.02 (d,J= 4.0 Hz, 1H), 7.82 - 7.74 (m, 2H), 7.65 (d,J= 3.9 Hz, 1H), 7.47 - 7.39 (m, 2H), 7.35 (t,J= 8.1 Hz , 1H), 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).13C NMR (75 MHz, DMSO-d6) 5 159.7, 157.7, 148.5, 138.4, 137.7, 134.3, 130.5, 130.4, 128.6 (2C), 127.4, 124.8, 121.8 (2C), 1 18.1, 116.1, 113.0, 73.5, 43.7 (2C), 32.3 (2C). HRMS (ESI) m/z: [M H]+ calc. for C<22>H<22>ClN<2>O<2>S 413.1085; found 413.1088. Purity (HPLC) > 99%.
N-(4-Clorofenil)-5-(4-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (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 CH<2>Cl<2>(3,2 mL) y ácido trifluoroacético (0,8 mL). Purificación: CH<2>Cl<2>/MeOH 9:1. Rendimiento: 45 mg, 55%. Sólido blanco. Pf: 240 - 242 °C. 1H RMN (300 MHz, DMSO-cfe) 510,40 (s, 1H), 8,04 (d,J= 4,0 Hz, 1H), 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). 13C RMN (75 MHz, DMSO-cfe) 5 159,8, 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 C<22>H<22>ClN<2>O<2>S 413,1085; encontrado 413,1080. Pureza (HPLC) > 99%.N-(4-Chlorophenyl)-5-(4-(piperidin-4-yloxy)phenyl)thiophene-2-carboxamide (27)Reagents: N-(4-chlorophenyl)-5-(4-hydroxyphenyl)thiophene-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, which were treated with a solution of CH<2>Cl<2>(3.2 mL) and trifluoroacetic acid (0.8 mL). Purification: CH<2>Cl<2>/MeOH 9:1. Yield: 45 mg, 55%. White solid. Mp: 240 - 242 °C. 1H NMR (300 MHz, DMSO-cfe) 510.40 (s, 1H), 8.04 (d,J= 4.0 Hz, 1H), 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). 13C NMR (75 MHz, DMSO-cfe) 5 159.8, 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), 41.5 (2C), 28.8 (2C). HRMS (ESI) m/z: [M H]+ calc. for C<22>H<22>ClN<2>O<2>S 413.1085; found 413.1080. Purity (HPLC) > 99%.
N-Fenetil-5-(2-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (28)N-Phenethyl-5-(2-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide (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 CH<2>Cl<2>(5 mL) y ácido trifluoroacético (3,3 mL). Purificación: CH<2>Ch/MeOH 9:1. Rendimiento: 30,5 mg, 16%. Sólido amarillo. Pf: descompone. 1H RMN (300 MHz, DMSO-da) 5 8,56 (t,J= 5,7 Hz, 1H), 7,78 (dd,J= 7,9, 1,7 Hz, 1H), 7,67 (d,J= 4,0 Hz, 1H), 7,59 (d,J= 4,0 Hz, 1H), 7,33 - 7,18 (m, 7H), 7,02 - 6,97 (m, 1H), 4,66 - 4,56 (m, 1H), 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).Reagents: N-phenethyl-5-(2-hydroxyphenyl)thiophene-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 CH<2>Cl<2>(5 mL) and trifluoroacetic acid (3.3 mL). Purification: CH<2>Ch/MeOH 9:1. Yield: 30.5 mg, 16%. Yellow solid. Pf: decomposes. 1H NMR (300 MHz, DMSO-da) 5 8.56 (t,J= 5.7 Hz, 1H), 7.78 (dd,J= 7.9, 1.7 Hz, 1H), 7.67 (d,J= 4.0 Hz, 1H), 7.59 (d,J= 4.0 Hz, 1H), 7.33 - 7.18 (m, 7H), 7.02 - 6.97 (m, 1H), 4.66 - 4.56 (m, 1H), 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-da) 5 161,5, 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]+ calc. para C<24>H<27>N<2>O<2>S 407,1788; encontrado 407,1782. Pureza (HPLC) > 99%.13C NMR (75 MHz, DMSO-da) 5 161.5, 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]+ calc. for C<24>H<27>N<2>O<2>S 407.1788; found 407.1782. Purity (HPLC) > 99%.
N-Fenetil-5-(3-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (29)N-Phenethyl-5-(3-(piperidin-4-yloxy)phenyl)thiophen-2-carboxamide (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.Reagents: N-phenethyl-5-(3-hydroxyphenyl)thiophene-2-carboxamide (19) (1.39 mmol, 450 mg), tert-butyl 4-hydroxypiperidine-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.
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 CH<2>Ch (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-de)58,62 (t,J= 5,6 Hz, 1H), 7,71 (d,J= 4,0 Hz, 1H), 7,54 (d,J= 3,9 Hz, 1H), 7,37 -7,16 (m, 8H), 6,96 (ddd,J= 8,2, 2,4, 1,1 Hz, 1H), 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). 13C RMN (75 MHz, DMSO-de) 5160,8, 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 (2C), 40,8, 35,2, 31,8 (2C). HRMS (ESI) m/z: [M H]+ calc. para C<24>H<27>N<2>O<2>S 407,1788; encontrado 407,1781. Pureza (HPLC): 96,6%.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 CH<2>Ch (5 mL) and trifluoroacetic acid (2.25 mL). Purification: no purification process was necessary. Yield: 145 mg, 79%. White solid. Mp: 152 - 154 °C. 1H NMR (300 MHz, DMSO-de)58.62 (t,J= 5.6 Hz, 1H), 7.71 (d,J= 4.0 Hz, 1H), 7.54 (d,J= 3.9 Hz, 1H), 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). 13C NMR (75 MHz, DMSO-de) 5160.8, 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 (2C), 40.8, 35.2, 31.8 (2C). HRMS (ESI) m/z: [M H]+ calc. for C<24>H<27>N<2>O<2>S 407.1788; found 407.1781. Purity (HPLC): 96.6%.
N-(4-Morfolinofenil)-5-(2-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (30)Reactivos: 5-(2-hidroxifenil)-N-(4-morfolinofenil)tiofen-2-carboxamida (20) (2,80 mmol, 786 mg), 4-hidroxipiperidin-1-carboxilato de terc-butilo (3,63 mmol, 729 mg), azodicarboxilato de diisopropilo (3,63 mmol, 714 pL), 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 terc-butilo que se trataron con una solución de CH<2>Ch (10 mL) y ácido trifluoroacético (6,6 mL). Purificación: la mezcla orgánica se acidificó con HCl 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-da) 510,05 (s, 1H), 7,96 (d,J= 4,1 Hz, 1H), 7,84 (dd,J= 7,9, 1,6 Hz, 1H,), 7,67 (d,J= 4,1 Hz, 1H), 7,63 - 7,56 (m, 2H), 7,34 (ddd,J= 8,7, 7,1, 1,6 Hz, 1H), 7,28 - 7,22 (m, 1H), 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).13C RMN (75 MHz, DMSO-da) 5159,7, 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), 114.2, 72,3, 66,1 (2C), 48,8 (2C), 42,2 (2C), 29,6 (2C). HRMS (ESI) m/z: [M H]+ calc. para C<26>^<0>^ O<3>S 464,2002; encontrado 464,2000. Pureza (HPLC) > 99%.N-(4-Morpholinophenyl)-5-(2-(piperidin-4-yloxy)phenyl)thiophene-2-carboxamide (30)Reagents: 5-(2-hydroxyphenyl)-N-(4-morpholinophenyl)thiophene-2-carboxamide (20) (2.80 mmol, 786 mg), tert-butyl 4-hydroxypiperidine-1-carboxylate (3.63 mmol, 729 mg), diisopropyl azodicarboxylate (3.63 mmol, 714 pL), triphenylphosphine (3.63 mmol, 951 mg) and 15 mL of THF. 300 mg of tert-butyl 4-(5-((4-morpholinophenyl)carbamoyl)thiophen-2-yl)phenoxy)piperidine-1-carboxylate were obtained, which were treated with a solution of CH<2>Ch (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 a yellow precipitate appeared. The precipitate was extracted with ethyl acetate. Yield: 57.6 mg, 24%. Yellow solid. Mp: 184 - 186 °C. 1H NMR (300 MHz, DMSO-da) 510.05 (s, 1H), 7.96 (d,J= 4.1 Hz, 1H), 7.84 (dd,J= 7.9, 1.6 Hz, 1H,), 7.67 (d,J= 4.1 Hz, 1H), 7.63 - 7.56 (m, 2H), 7.34 (ddd ,J= 8.7, 7.1, 1.6 Hz, 1H), 7.28 - 7.22 (m, 1H), 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) 52.7, 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), 114.2, 72.3, 66.1 (2C), (2C), 42.2 (2C), 29.6 (2C). HRMS (ESI) m/z: [M H]+ calc. for C<26>^<0>^ O<3>S 464,2002; found 464,2000. Purity (HPLC) > 99%.
Síntesis de M-fenil-5-(2-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (31)Sobre una mezcla de(8)5-bromo-N-feniltiofen-2-carboxamida (1 equiv.), Na<2>CO<3>(2,2 equiv.) y paladio (0) tetrakis(trifenilfosfina) (0,05 equiv.), se añadió una disolución de tolueno, H<2>O 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 120 °C. Finalizado este tiempo, el crudo de reacción se adicionó una disolución 1:1 de AcOEt/H<2>O y se extrajo con AcOEt. La fase orgánica se lavó con una disolución 1:1 de disolución saturada de NaCl y H<2>O, y finalmente se secó sobre Na<2>SO<4>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.Synthesis of M-phenyl-5-(2-(piperidin-4-yloxy)phenyl)thiophene-2-carboxamide (31)To a mixture of (8)5-bromo-N-phenylthiophene-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 2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (1.2 equiv.) was added. The reaction mixture was bubbled with argon for 15 min and then heated under microwave (MW) irradiation for 30 min at 120 °C. After this time, the reaction crude was added 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 and H<2>O, and finally dried over anhydrous Na<2>SO<4>. The desiccant was then 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 °C se añadió DIAD (1,3 equiv.) y 4-hidroxipiperidina-1-carboxilato de terc-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 CH<2>CL/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 CH<2>CE se basificó con NaHCO<3>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 CH<2>Cl<2>/MeOH 9:1. Rendimiento: 90,5 mg, 35%. Sólido blanco. Pf: 122 - 124 °C. 1H RMN (300 MHz, DMSO-da) 510,28 (s, 1H), 8,07 (d,J= 4,1 Hz, 1H), 7,87 (dd,J= 7,9, 1,6 Hz, 1H), 7,81 - 7,73 (m, 2H), 7,70 (d,J= 4,1 Hz, 1H), 7,41 - 7,31 (m, 3H), 7,29 (d,J= 7,7 Hz, 1H), 7,12 (d,J= 7,5 Hz, 1H), 7,07 (dd,J= 7,8, 1,2 Hz, 1H), 4,91 (dt,J= 7,9, 4,0 Hz, 1H), 3,30 (m, 2H), 3,13 (m, 2H), 2,21 (m, 2H), 1,98 (m, 2H). 13C RMN (75 MHz, DMSO-de) 5160,5, 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]+ calc. para C<22>H<23>N<2>O<2>S 379,1475; encontrado 379,1473. Pureza (HPLC) > 99%.To a solution of the above residue (1 equiv.) and triphenylphosphine (1.3 equiv.) in THF at 0 °C was added DIAD (1.3 equiv.) and tert-butyl 4-hydroxypiperidine-1-carboxylate (1.3 equiv.). The mixture was left stirring overnight and then the solvent was removed under reduced pressure. The white solid residue obtained was dissolved in a 3:2 CH<2>CL/TFA solution and stirred at room temperature for 3 hours. After deprotection was complete, the solvent was evaporated in vacuo and after dissolving the resulting product in CH<2>CE 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. Mp: 122 - 124 °C. 1H NMR (300 MHz, DMSO-da) 510.28 (s, 1H), 8.07 (d,J= 4.1 Hz, 1H), 7.87 (dd,J= 7.9, 1.6 Hz, 1H), 7.81 - 7.73 (m, 2H), 7.70 (d,J= 4.1 Hz, 1H), 7.41 - 7, 31 (m, 3H), 7.29 (d,J= 7.7 Hz, 1H), 7.12 (d,J= 7.5 Hz, 1H), 7.07 (dd,J= 7.8, 1.2 Hz, 1H), 4.91 (dt,J= 7.9, 4.0 Hz, 1H), 3.30 (m, 2H), 3.13 (m, 2H), 2.21 (m, 2H), 1.98 (m, 2H). 13C NMR (75 MHz, DMSO-de) 5160.5, 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]+ calc. for C<22>H<23>N<2>O<2>S 379.1475; found 379.1473. Purity (HPLC) > 99%.
Síntesis de N-fenil-5-(3-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (32)Sobre una mezcla de(8)5-bromo-N-feniltiofen-2-carboxamida (1 equiv.), Na<2>CO<3>(2,2 equiv.) y paladio (0) tetrakis(trifenilfosfina) (0,05 equiv.), se añadió una disolución de tolueno, H<2>O 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 °C. Finalizado este tiempo, el crudo de reacción se adicionó una disolución 1:1 de AcOEt/H<2>O y se extrajo con AcOEt. La fase orgánica se lavó con una disolución 1:1 de disolución saturada de NaCl y H<2>O, y finalmente se secó sobre Na<2>SO<4>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.Synthesis of N-phenyl-5-(3-(piperidin-4-yloxy)phenyl)thiophene-2-carboxamide (32)To a mixture of (8)5-bromo-N-phenylthiophene-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 (1.2 equiv.) was added. The reaction mixture was bubbled with argon for 15 min and then heated under microwave (MW) irradiation for 30 min at 120 °C. After this time, the reaction crude was added 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 and H<2>O, and finally dried over anhydrous Na<2>SO<4>. The desiccant was then 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 °C se añadió DIAD (1,3 equiv.) y 4-hidroxipiperidina-1-carboxilato de terc-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 CH<2>Cl<2>/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 CH<2>Cl<2>se basificó con NaHCÜ<3>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 CH<2>Cl<2>/MeOH 9:1. Rendimiento: 80,8 mg, 32%. Sólido blanco. Pf: 151 - 153 °C. 1H RMN (300 MHz, DMSO-da) 510,27 (s, 1H), 8,05 (d,J= 4,0 Hz, 1H), 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, 1H), 4,56 (dt,J= 8,8, 4,7 Hz, 1H), 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-Ó6) 5160,0, 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]+ calc. para C<22>H<23>N<2>O<2>S 379,1475; encontrado 379,1475. Pureza (HPLC) > 99%.To a solution of the above residue (1 equiv.) and triphenylphosphine (1.3 equiv.) in THF at 0 °C was added DIAD (1.3 equiv.) and tert-butyl 4-hydroxypiperidine-1-carboxylate (1.3 equiv.). The mixture was left stirring overnight and then the solvent was removed under reduced pressure. The white solid residue obtained was dissolved in a 3:2 CH<2>Cl<2>/TFA solution and stirred at room temperature for 3 hours. After deprotection was complete, the solvent was evaporated in vacuo and after dissolving the resulting product in CH<2>Cl<2> it was basified with NaHCÜ<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: 80.8 mg, 32%. White solid. Mp: 151 - 153 °C. 1H NMR (300 MHz, DMSO-da) 510.27 (s, 1H), 8.05 (d,J= 4.0 Hz, 1H), 7.81 - 7.70 (m, 2H), 7.65 (d,J= 4.0 Hz, 1H), 7.36 (m, 3H), 7.28 (m, 2H), 7.21 - 7.0 5 (m, 1H), 6.99 (d,J= 8.1 Hz, 1H), 4.56 (dt,J= 8.8, 4.7 Hz, 1H), 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 NMR (75 MHz, DMSO-Ó6) 5160.0, 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, , 73.2, 43.5, 31.9. HRMS (ESI) m/z: [M H]+ calc. for C<22>H<23>N<2>O<2>S 379.1475; found 379.1475. Purity (HPLC) > 99%.
Síntesis de N-fenil-5-(4-(piperidin-4-iloxi)fenil)tiofen-2-carboxamida (33)Sobre una mezcla de(8)5-bromo-N-feniltiofen-2-carboxamida (1 equiv.), Na<2>CO<3>(2,2 equiv.) y paladio (0) tetrakis(trifenilfosfina) (0,05 equiv.), se añadió una disolución de tolueno, H<2>O 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 120 °C. Finalizado este tiempo, el crudo de reacción se adicionó una disolución 1:1 de AcOEt/H<2>O y se extrajo con AcOEt. La fase orgánica se lavó con una disolución 1:1 de disolución saturada de NaCl y H<2>O, y finalmente se secó sobre Na<2>SO<4>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.Synthesis of N-phenyl-5-(4-(piperidin-4-yloxy)phenyl)thiophene-2-carboxamide (33)To a mixture of (8) 5-bromo-N-phenylthiophene-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 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (1.2 equiv.) was added. The reaction mixture was bubbled with argon for 15 min and then heated under microwave (MW) irradiation for 30 min at 120 °C. After this time, the reaction crude was added 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 and H<2>O, and finally dried over anhydrous Na<2>SO<4>. The desiccant was then 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 °C se añadió DIAD (1,3 equiv.) y 4-hidroxipiperidina-1-carboxilato de terc-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 CH<2>CI<2>/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 CH<2>Cl<2>se basificó con NaHCO<3>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 CH<2>Cl<2>/MeOH 9:1. Rendimiento: 96,3 mg, 38%. Sólido blanco. Pf: 192 - 194 °C. 1H RMN (300 MHz, DMSO-da) ó 10,22 (s, 1H), 8,01 (d,J= 4,0 Hz, 1H), 7,79 - 7,68 (m, 2H), 7,68 - 7,57 (m, 2H), 7,49 (d,J= 3,9 Hz, 1H), 7,36 (t,J= 7,9 Hz, 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). 13C RMN (75 MHz, DMSO da) 5 160,1, 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 C<22>H<23>N<2>O<2>S 379,1475; encontrado 379,1469. Pureza (HPLC) > 99%.To a solution of the above residue (1 equiv.) and triphenylphosphine (1.3 equiv.) in THF at 0 °C was added DIAD (1.3 equiv.) and tert-butyl 4-hydroxypiperidine-1-carboxylate (1.3 equiv.). The mixture was left stirring overnight and then the solvent was removed under reduced pressure. The white solid residue obtained was dissolved in a 3:2 CH<2>Cl<2>/TFA solution and stirred at room temperature for 3 hours. After deprotection was complete, the solvent was evaporated in vacuo and after dissolving the resulting product in CH<2>Cl<2>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: 96.3 mg, 38%. White solid. Mp: 192 - 194 °C. 1H NMR (300 MHz, DMSO-da) or 10.22 (s, 1H), 8.01 (d,J= 4.0 Hz, 1H), 7.79 - 7.68 (m, 2H), 7.68 - 7.57 (m, 2H), 7.49 (d,J= 3.9 Hz, 1H), 7.36 (t,J= 7.9 Hz , 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). 13C NMR (75 MHz, DMSO da) 5 160.1, 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, 3 1.9. HRMS (ESI) m/z: [M H]+ calc. for C<22>H<23>N<2>O<2>S 379.1475; found 379.1469. Purity (HPLC) > 99%.
Síntesis de N-(2-(1-bencilpiperidin-4-il)etil)-5-(3-metoxifenil)tiofen-2-carboxamida (34)Synthesis of N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-(3-methoxyphenyl)thiophen-2-carboxamide (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 °C, 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 terc-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 CH<2>Ch/MeOH 9:1. Rendimiento: 80,1 mg, 12 %. Sólido blanco. Pf: 99 - 101 °C. 1H RMN (300 MHz, DMSO-d6) 58,46 (t,J= 5,6 Hz, 1H), 7,72 (d,J= 4,0 Hz, 1H), 7,54 (d,J= 3,9 Hz, 1H), 7,39 - 7,18 (m,<8>H), 6,94 (ddd,J= 8,2, 2,5, 1,0 Hz, 1H), 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, 1H), 1,23 - 1,04 (m, 2H).To a solution of triphenylphosphine (2.07 mmol, 542 mg) in THF at 0 °C was added DIAD (2.07 mmol, 408 pL). 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)thiophene-2-carboxamide (10) (1.59 mmol, 669 mg), tert-butyl 4-hydroxypiperidine-1-carboxylate (2.07 mmol, 415 mg) was added and stirred overnight. The solvent was then removed under reduced pressure and the resulting residue was purified by column chromatography using a CH<2>Ch/MeOH 9:1 mixture as eluent. Yield: 80.1 mg, 12%. White solid. Mp: 99 - 101 °C. 1H NMR (300 MHz, DMSO-d6) 58.46 (t,J= 5.6 Hz, 1H), 7.72 (d,J= 4.0 Hz, 1H), 7.54 (d,J= 3.9 Hz, 1H), 7.39 - 7.18 (m,<8>H), 6.94 (ddd,J= 8.2, 2.5, 1.0 Hz, 1H), 3.81 (s, 3H), 3.43 (s, 2H), 3.26 (q,J= 6.7 Hz, ), 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, 1H), 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]+ calc. para C<26>H<31>N<2>O<2>S 435,2101; encontrado 435,2099. Pureza (HPLC): 97,7 %.13C NMR (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]+ calc. for C<26>H<31>N<2>O<2>S 435.2101; found 435.2099. Purity (HPLC): 97.7%.
Síntesis de N-(2-(1-bencilpiperidin-4-il)etil)-5-(4-metoxifenil)tiofen-2-carboxamida (35)Synthesis of N-(2-(1-benzylpiperidin-4-yl)ethyl)-5-(4-methoxyphenyl)thiophen-2-carboxamide (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 0 °C, 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 terc-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 mezcla CH<2>Ch/MeOH 9:1. Rendimiento: 70,3 mg, 16 %. Sólido beige. Pf: 163 - 165 °C. 1H RMN (300 MHz, DMSO-cfe) ó 8,41 (t,J= 5,6 Hz, 1H), 7,68 (d,J= 3,9 Hz, 1H), 7,66 - 7,58 (m, 2H), 7,38 (d,J= 3,9 Hz, 1H), 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). 13C RMN (75 MHz, DMSO-cfe) 5160,9, 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 C<26>H<31>N<2>O<2>S 435,2101; encontrado 435,2102. Pureza (HPLC): 96,4 %.To a solution of triphenylphosphine (1.15 mmol, 334 mg) in THF at 0 °C was added DIAD (1.15 mmol, 266 pL). 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)thiophene-2-carboxamide (11) (1.04 mmol, 438 mg) and tert-butyl 4-hydroxypiperidine-1-carboxylate (1.15 mmol, 230 mg) in THF was added and stirred overnight. The solvent was then removed under reduced pressure and the resulting residue was purified by column chromatography using a CH<2>Ch/MeOH 9:1 mixture as eluent. Yield: 70.3 mg, 16 %. Beige solid. Mp: 163 - 165 °C. 1H NMR (300 MHz, DMSO-cfe) or 8.41 (t,J= 5.6 Hz, 1H), 7.68 (d,J= 3.9 Hz, 1H), 7.66 - 7.58 (m, 2H), 7.38 (d,J= 3.9 Hz, 1H), 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). 13C NMR (75 MHz, DMSO-cfe) 5160.9, 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 C<26>H<31>N<2>O<2>S 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 (EBOV)Example 2: Study of the antiviral activity of the compounds of formula (I) against the Ebola virus (EBOV)
Materiales y MétodosMaterials and methods
Líneas celularesCell 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 africanoCercopithecus 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 CO<2>.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 African green monkey kidney (Cercopithecus aethiops) cells (VeroE6) were cultured in DMEM medium supplemented with 10% FBS, 1% L-glutamine and 25 pg/mL gentamicin in a 37 °C incubator in the presence of 5% CO<2>.
Producción de virus recombinantes (HIV-May-luc) basados en el virus de inmunodeficiencia humana (VIH), con envolturas (GP/G) de virus Ebola (ZEBOV) Mayinga o Virus de la Estomatitis Vesicular (VSV) y que expresan luciferasa 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) Mayinga or Vesicular Stomatitis Virus (VSV) and which express luciferase 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.The production of these viral particles was carried out using the calcium chloride transfection method (Life Technologies, Carlsbad, CA, USA) in 293T cells by adding the plasmids: pNL4.3.Luc.RE- (NIH AIDS Reagent Program, Division of AIDS of Dr. Nathaniel Landau) and the respective glycoprotein of each 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 -80 °C en alícuotas.Supernatants containing recombinant viruses were collected 48 h after transfection, centrifuged at 1200 rpm for 10 min at room temperature to remove cell 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, WI) 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 post-infection by luciferase assay (Luciferase Assay System, Promega, Madison, WI) using the GloMax® Navigator Microplate Luminometer (Promega) for reading.
Producción de virus recombinantes (VSV-Eb-luc) basados en el virus de la Estomatitis Vesicular (VSV), con envolturas (GP/G) de virus Ebola (ZEBOV) Mayinga 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 Vesicular Stomatitis Virus (VSV), with envelopes (GP/G) of Ebola virus (ZEBOV) Mayinga or Vesicular Stomatitis Virus (VSV) and which express 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 vesicular stomatitis virus-based pseudotypes, BHK-21 cells (Kerafast) were transfected with plasmids encoding the Ebola virus envelope protein (Mayinga strain) or the vesicular stomatitis virus glycoprotein (G) itself using Lipofectamine 3000 (Thermo Fisher Scientific). After 24 h, the cells were inoculated with recombinant VSV-delta G-luc pseudotypes, deficient in their replication capacity (MOI: 3-5), containing the firefly luciferase gene replacing the virus's own glycoprotein, thus allowing the incorporation of heterologous glycoproteins into the new viral particles that were generated.
Tras 1 h de incubación a 37 °C, se retira el inóculo, se lavan varias veces las células con PBS y finalmente se añade medio fresco.After 1 h of incubation at 37 °C, 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 °C.Viral particles are collected after 20-24 h post-inoculation, cellular debris is removed by centrifugation (1200 rpm, 10 minutes) and stored in aliquots in a -80 °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-Glo® Luciferase Assay System, Promega) utilizando para la lectura el luminómetro GloMax® Navigator Microplate Luminometer (Promega).The infectious titer (infectious dose per milliliter) was calculated 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-Glo® Luciferase Assay System, Promega) using the GloMax® Navigator Microplate Luminometer (Promega) for reading.
Cribado de compuestos inhibidores de la infección por virus ÉbolaScreening 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 pocillos en presencia de cada compuesto a una concentración final de 10 pM.Screening for Ebola virus entry inhibitor compounds 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 °C 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, WI, EE. UU.) con el sistema de ensayo de luficerasa (Promega, Madison, WI).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, WI).
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 virus VSV-G 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 reported 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 (CI<50>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 further analysis (IC<50> and cytotoxicity).
Cálculo de los valores de CI<50>Calculation of CI<50> values
Para calcular el valor CI<50>, se emplearon concentraciones seriadas de cada compuesto desde 10 pM a 10 nM.To calculate the IC<50> 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 dosisrespuesta.Inhibition values were calculated using the mean 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.
Análisis de toxicidad de compuestosToxicity analysis of compounds
Se emplearon células Hela (2x104) tratadas con diferentes concentraciones: 10, 100 y 250 pM de cada compuesto durante 48 horas en placa de 96 pocillos. 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 (2x104) were treated with different concentrations: 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 by 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 °C en una atmósfera humidificada de CO<2>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 µL of the combined MTS/PMS solution (reagent) was mixed with 100 µL of cells in culture medium and further incubated for 2 h at 37 °C in a humidified 5% CO<2> atmosphere. The 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.
ResultadosResults
aCÍ<5>o: Concentración inhibitoria 50% o requerida para reducir la infección viral en un 50% en los pseudotipos indicados en cada caso.aCÍ<5>o: 50% inhibitory concentration or required to reduce viral infection by 50% in the pseudotypes indicated in each case.
bCC<50>: Concentración citotóxica 50% o requerida para reducir el crecimiento celular en un 50% en el tipo de células indicados en cada casobCC<50>: Cytotoxic concentration 50% or required to reduce cell growth by 50% in the cell type indicated in each case
cIS: Índice de selectividad (CC<50>/CI<50>)cIS: Selectivity index (CC<50>/CI<50>)
Ejemplo 3: Estudio de la actividad antiviral de los compuestos de fórmula (I) frente al virus de la peste porcina africana (VPPA)Example 3: Study of the antiviral activity of the compounds of formula (I) against the African swine fever virus (ASFV)
Materiales y MétodosMaterials and methods
Tratamiento e infecciónTreatment 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 Ba71V 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 Virol32, 471-477) o con los virus recombinantes fluorescentes para citometría de flujo como se describe más abajo.For the performance of the various assays, ASFV infections were performed on Vero cell monolayers (ATCC-CCL-81; renal fibroblasts) at a multiplicity of infection (moi) of 5 pfu/cell of the non-pathogenic ASFV isolate Ba71V 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 the 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 37 °C. 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).Viral inocula were always added to the minimum volume necessary to cover the cell wall, after pretreatment of the cells with the compounds for one hour at 37 °C. After the pretreatment time, the viral inoculum was allowed to absorb for 60 minutes at 37 °C and finally, the medium was removed together with the viral inoculum and replaced with Dulbecco's modified Eagle's medium (DMEM) supplemented with 2% fetal bovine serum (FBS) in combination with the different compounds to continue with 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. They were then diluted in 2% DMEM medium in fetal bovine serum (FBS) until reaching the required concentration in each case.
Ensayos de CitotoxicidadCytotoxicity 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 absorbancia 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 a 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 in formazan by the activity of dehydrogenase enzymes of metabolically active cells.
Detección y cuantificación del DNA viralDetection 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).Following infection and treatment with the different compounds, the cells were extracted and DNA was subsequently obtained and purified using the "Dneasy blood and tissue kit” (Quiagen). The concentrations obtained were quantified using 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 Methods2003,107,53-61). 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 Kinget al., obtenidos de Fisher Scientific y "Premix Ex Taq (2X)” (Takara).For the analysis of viral replication, quantitative real-time PCR (qPCR) was performed using fluorescent hybridization probes aimed at amplifying the viral p72 gene region as previously described (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). For this purpose, the previously purified DNA samples were analyzed, using water as a negative control, samples treated with DMSO as a positive control and as a standard curve, 5 serial dilutions of purified viral DNA of known concentration. The amplification mixture consisted of 150 ng of DNA from each of the samples, the Taqman probe (Roche), the primers OE3F and OE4R whose sequence is described by Kinget 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 94 °C durante 10 minutos, 45 ciclos de 94 °C durante 15 segundos y 58 °C durante 60 segundos.The amplification and quantification process was carried out in an "ABI 7500 Fast Real-Time PCR System” thermal cycler (Applied Biosystems) with the following parameters: 1 cycle of 94 °C for 10 minutes, 45 cycles of 94 °C for 15 seconds and 58 °C for 60 seconds.
Citometría de flujoFlow cytometry
Para el análisis por citometría de flujo sin el marcaje 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 One2017,12,e018974 y en Hernaez, 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.Virology2006,350,1-14, 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, “fluorescenceactivated cell sorter”). Finalmente se analizaron 10000 células por experimento en un citómetro 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 CI<50>empleando diferentes concentraciones de compuestos.For flow cytometric analysis without antibody labeling, cells were infected with 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 One2017,12,e018974 and in Hernaez, 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.Virology2006,350,1-14, 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 cytometer (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 the results were obtained with the optimal concentrations, the IC<50> were calculated using different concentrations of compounds.
Análisis estadísticoStatistical 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 GraphPad 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 the control values. Statistical analysis of the variance between data was performed using GraphPad Prism 6 software, applying the Bonferroni test for multiple comparisons.
ResultadosResults
aCÍ<5>o: Concentración inhibitoria 50% o requerida para reducir la infección viral en un 50%.aCÍ<5>o: Inhibitory concentration 50% or required to reduce viral infection by 50%.
bCC<50>: Concentración citotóxica 50% o requerida para reducir el crecimiento celular en un 50%.bCC<50>: Cytotoxic concentration 50% or required to reduce cell growth by 50%.
cIS: Índice de selectividad (CC<50>/CI<50>).cIS: Selectivity index (CC<50>/CI<50>).
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2019036562A1 (en)* | 2017-08-18 | 2019-02-21 | Saint Louis University | Err inverse agonists |
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2019036562A1 (en)* | 2017-08-18 | 2019-02-21 | Saint Louis University | Err inverse agonists |
| Title |
|---|
| AURORA FINE CHEMICALS (LIBRERÍA QUÍMICA). RN 890826-75-2. Base de datos: REGISTRY , 06/07/2006, Recuperado de Internet (URL:STN Internacional),* |
| AURORA FINE CHEMICALS (LIBRERÍA QUÍMICA). RN 922436-24-6. Base de datos: REGISTRY , 22/02/2007, Recuperado de Internet (URL:STN Internacional),* |
| AURORA FINE CHEMICALS (LIBRERÍA QUÍMICA). RN 923517-02-6. Base de datos: REGISTRY, 27/02/2007,* |
| AURORA FINE CHEMICALS (LIBRERÍA QUÍMICA). RN 923531-76-4. REGISTRY, 27/02/2007, Recuperado de Internet (URL:STN Internacional),* |
| AURORA FINE CHEMICALS (LIBRERÍA QUÍMICA). RN 923537-15-9. Base de datos: REGISTRY , 27/02/2007,* |
| AURORA FINE CHEMICALS (LIBRERÍA QUÍMICA). RN 923543-36-6. Base de datos: REGISTRY , 27/02/2007,* |
| BIONET RESEARCH LTD. (LIBRERÍA QUÍMICA). RN 344263-75-8. Base de datos: REGISTRY , 01/07/2001, Recuperado de Internet (URL:STN Internacional),* |
| PASCUAL, M. J. ET AL., . Structure-based drug design for envelope protein E2 uncovers a new class of bovine viral diarrhea inhibitors that block virus entry. Antiviral Research, 12/10/2017, Vol. 149, Páginas 179-190 (DOI: 10.1016/j.antiviral.2017.10.010) Ver resumen y esquema 1.* |
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|---|---|
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| Publication | Publication Date | Title |
|---|---|---|
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