ES2600955T3 - Adenovirus oncolíticos para el tratamiento del cáncer - Google Patents

Adenovirus oncolíticos para el tratamiento del cáncer Download PDF

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ES2600955T3
ES2600955T3 ES10772050.0T ES10772050T ES2600955T3 ES 2600955 T3 ES2600955 T3 ES 2600955T3 ES 10772050 T ES10772050 T ES 10772050T ES 2600955 T3 ES2600955 T3 ES 2600955T3
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adenovirus
oncolytic adenovirus
oncolytic
tumor
cancer treatment
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ES2600955T5 (en
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Sònia GUEDAN CARRIÓ
Manel Maria Cascallo Piqueras
Ramón ALEMANY BONASTRE
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Fundacio Privada Inst D'investigacio Biomedica
Fundacio Privada Institut D'investigacio Biomedica
Institut Catala dOncologia
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Fundacio Privada Institut D'investigacio Biomedica
Institut Catala dOncologia
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Abstract

La invención se refiere a un adenovirus oncolítico que comprende una secuencia que codifica una enzima hialuronidasa insertada en su genoma. Este adenovirus se distribuye más eficientemente por la masa tumoral y por consiguiente se aumenta el efecto oncolítico. Inyectando el adenovirus oncolítico de la invención endovenosamente se obtienen regresiones del volumen tumoral. Por lo tanto el adenovirus oncolítico de la presente invención es útil para el tratamiento del cáncer o de un estado pre-maligno del mismo.

Description

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adenovírica a modificar, y después realizar una recombinación homóloga en bacterias con un plásmido que contiene el resto del genoma vírico.
El adenovirus que contiene el gen de la hialuronidasa objeto de la presente invención se propaga y amplifica en
5 líneas celulares normalmente utilizadas en el campo de la terapia génica y viroterapia, tales como las líneas HEK-293 y A549. El método preferente de propagación es por infección de una línea celular permisiva a la replicación del adenovirus. La línea de adenocarcinoma pulmonar A549 es un ejemplo de una línea con tales características. La propagación se realiza por ejemplo del siguiente modo: las células A549 se siembran en placas de cultivo celular de plástico y se infectan utilizando 100 partículas víricas por célula. Dos días después el efecto
10 citopático que refleja la producción de virus se observa como un agrupamiento y redondamiento de las células. Las células se recogen y se almacenan en tubos. Después de una centrifugación a 1000 g durante 5 minutos, el sedimento celular se congela y descongela tres veces para romper las células. El extracto celular resultante se centrifuga a 1000 g durante 5 minutos y el sobrenadante con virus se carga en un gradiente de cloruro de cesio y se centrifuga durante 1 hora a 35000 g. La banda de virus obtenida del gradiente se carga de nuevo en otro
15 gradiente de cloruro de cesio y se centrifuga otra vez durante 16 horas a 35000 g. La banda de virus se recoge y se dializa frente a PBS-glicerol al 10%. El virus dializado se alícuota y mantiene a -80 ºC. La cuantificación del número de partículas víricas y de unidades formadoras de placa se realiza siguiendo protocolos convencionales. La solución salina tamponada con fosfato (PBS) con glicerol al 5% es una formulación convencional para el almacenamiento de adenovirus. No obstante, se han descrito nuevas formulaciones que mejoran la estabilidad del
20 virus. Los métodos de purificación del adenovirus que contiene el gen de la hialuronidasa para su utilización en el tratamiento del cáncer, son los mismos que los descritos para otros adenovirus y vectores adenovíricos utilizados en viroterapia y terapia génica del cáncer.
El adenovirus oncolítico de la presente invención puede administrarse a un mamífero, preferentemente un humano.
25 La intención de la administración del adenovirus oncolítico es terapéutica, incluyendo, pero sin limitación, melanoma, cáncer de páncreas, cáncer de colon y cáncer de pulmón. Además, se contempla la administración del adenovirus oncolítico en una fase pre-maligno de un tumor.
Se entiende que el adenovirus oncolítico se administra en una forma farmacéuticamente aceptable. Los expertos
30 en la materia pueden cerciorarse de la dosis apropiada utilizando procedimientos convencionales. Se entiende que la dosis debe ser una cantidad eficaz del adenovirus oncolítico para que produzca una reducción del tumor en el paciente tratado. El virus se puede administrar de forma directa virus al tumor, en la cavidad donde se emplaza el tumor, en la vasculatura del tumor, alrededor del tumor o por la inyección endovenosa sistémica en el paciente. Preferentemente, la administración es sistémica.
35 Los protocolos para utilizar los virus descritos en la presente invención para el tratamiento del cáncer son los mismos procedimientos utilizados en los campos de la viroterapia con adenovirus y terapia génica con adenovirus. Existe una amplia experiencia en el uso de adenovirus no oncolíticos y oncolíticos en el campo de la terapia génica. Existen numerosas publicaciones que describen el tratamiento de células tumorales en cultivo, en modelos
40 animales y en ensayos clínicos con pacientes. Para el tratamiento de células en cultivos in vitro el adenovirus purificado mediante cualquiera de las formulaciones descritas anteriormente, se añade al medio de cultivo para obtener la infección de las células tumorales. Para tratar tumores en modelos animales o en pacientes humanos, el adenovirus se puede administrar loco-regionalmente mediante inyección en el tumor o en una cavidad corporal donde se localiza el tumor, o bien por vía sistémica mediante inyección en la circulación sanguínea.
45 El adenovirus oncolítico de la invención puede administrarse solo o en una composición con transportadores o excipientes farmacéuticamente aceptables. El experto en la materia adaptará la composición de acuerdo con el modo particular de administración. Las composiciones pueden comprender el adenovirus oncolítico como el único agente contra el tumor, o en combinación con otro agente terapéutico tal como un fármaco quimioterapéutico o un
50 vector con un gen terapéutico insertado. También puede combinarse la terapia con el adenovirus oncolítico con radioterapia.
A menos que se defina de otro modo, todos los términos técnicos y científicos utilizados en el presente documento tienen el mismo significado a los que entiende comúnmente una persona experta en la materia. En la práctica de la 55 presente invención pueden utilizarse métodos y materiales similares o equivalentes a los descritos en el presente documento. A lo largo de la descripción y las reivindicaciones la palabra "comprende" y sus variantes no pretenden excluir otras características técnicas, aditivos, componentes o etapas. Los objetivos, ventajas y características adicionales de la invención, serán evidentes para los expertos en la materia después de examinar la descripción o pueden aprenderse mediante la práctica de la invención. Las siguientes realizaciones particulares y los dibujos se
60 proporcionan a modo de ilustración, y no se pretende que sean limitativos de la presente invención.
Descripción de los dibujos
La FIG. 1 (a) muestra la estructura de adenovirus oncolíticos caracterizados por contener y expresar el gen de la
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Claims (1)

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ES10772050T 2009-05-06 2010-05-05 Oncolytic adenoviruses for treating cancer Active ES2600955T5 (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
ES200901201 2009-05-06
ES200901201A ES2385251B1 (es) 2009-05-06 2009-05-06 Adenovirus oncolíticos para el tratamiento del cáncer.
PCT/ES2010/000196 WO2010128182A1 (es) 2009-05-06 2010-05-05 Adenovirus oncolíticos para el tratamiento del cáncer

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ES2600955T3 true ES2600955T3 (es) 2017-02-13
ES2600955T5 ES2600955T5 (en) 2025-10-02

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ES200901201A Expired - Fee Related ES2385251B1 (es) 2009-05-06 2009-05-06 Adenovirus oncolíticos para el tratamiento del cáncer.
ES10772050T Active ES2600955T5 (en) 2009-05-06 2010-05-05 Oncolytic adenoviruses for treating cancer

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US (3) US10316065B2 (es)
EP (1) EP2428229B2 (es)
JP (1) JP2012525833A (es)
KR (1) KR101878274B1 (es)
CN (2) CN102548584A (es)
AU (1) AU2010244348B2 (es)
BR (1) BRPI1011445B8 (es)
CA (1) CA2761183C (es)
ES (2) ES2385251B1 (es)
FI (1) FI2428229T4 (es)
HK (1) HK1244679A1 (es)
MX (1) MX2011011818A (es)
PL (1) PL2428229T5 (es)
RU (1) RU2536931C2 (es)
WO (1) WO2010128182A1 (es)

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