ES2542059T3 - Nuevos anticuerpos que inhiben la dimerización de c-Met y sus utilizaciones - Google Patents
Nuevos anticuerpos que inhiben la dimerización de c-Met y sus utilizaciones Download PDFInfo
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- ES2542059T3 ES2542059T3 ES08786050.8T ES08786050T ES2542059T3 ES 2542059 T3 ES2542059 T3 ES 2542059T3 ES 08786050 T ES08786050 T ES 08786050T ES 2542059 T3 ES2542059 T3 ES 2542059T3
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- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
- C07K16/30—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells
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- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/32—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against translation products of oncogenes
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- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/40—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against enzymes
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- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
- G01N33/57492—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites involving compounds localized on the membrane of tumor or cancer cells
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- C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding
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- C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
- C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
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Abstract
Anticuerpo aislado, o uno de sus fragmentos divalentes de unión a c-Met, caracterizado por que el anticuerpo comprende una cadena pesada que comprende CDR-H1, CDR-H2 y CDR-H3 que comprenden respectivamente la secuencia de aminoácidos SEC ID nº 1, 2 y 3; y una cadena ligera que comprende CDR-L1, CDR-5 L2 y CDR-L3 que comprenden respectivamente la secuencia de aminoácidos SEC ID nº 10, 11 y 12.
Description
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retiraron suavemente con una torunda de algodón, células que migraron a la superficie inferior del filtro se lisaron, tiñeron con amortiguador de tinción CyQuant GR (Invitrogen) y contaron utilizando un lector de fluorescencia Berthold Mithras LB940. Todas las condiciones se probaron como triplicados.
Como se esperó HGF induce una invasión significante de células de tumor, comparables con la observada con FCS al 10% introducido como un control positivo (Figura 8). El IgG1 9G4 murino introducido como un control de isotipo no tiene efecto significante en invasión inducida HGF o basal, cuando se compara con células revestidas sin IgG. No se notó efecto agonista con 11E1, 224G11, 223C4 y 227H1 cuando se agregan solos y una inhibición significante y comparable de la invasión inducida por HGF se observa con los 3 Mabs.
Ejemplo 6: Inhibición de sanación de heridas por anticuerpos anti-c-Met
HGF estimula la movilidad. Para determinar si los anticuerpos anti-HGF fueron capaces de inhibir migración, células NCI-H441 se desarrollaron a alta densidad y se introdujo un espacio con una punta de pipeta P200. Las células después se estimularon para migrar a través del espacio con HGF (100 ng/ml), en presencia o ausencia de 11E1. pocillos con 11E1 solos, fueron también evaluados. Cada condición de prueba se evaluó como un sextuplicado y 3 experimentos independientes se realizaron. Después de una incubación durante la noche, las células se visualizaron con una Axio Vision Camera (objetivo x4).
HGF induce una migración significante resultando en un cierre completo del espacio dentro de una noche (Figura 9). IgG1 irrelevante 9G4 empleado como un control de isotipo no tiene efecto alguno en migración celular. Como se esperó, se observó un efecto agonista con 5D5 cuando se agrega solo pero una inhibición significante de migración celular se observa con este anticuerpo en presencia de HGF en la porción del espacio que permanece abierto. El fragmento Fab de 5D5 está sin efecto agonista cuando se agrega solo. Sin embargo, no se observa actividad de este fragmento en la presencia de HGF. Como se observa con el control de isotipo 9G4, MAb 11E1 no tuvo efecto agonista cuando se agrega solo y se comporta como un antagonista completo en la presencia de HGF.
Ejemplo 7: Ensayo de Dispersión
Células SK-HEP-1 se sembraron a baja densidad (1.104 células/pozo) en una placa de 24-pozos en DMEM con FCS al 10% y desarrollan por 24 horas antes de adición, al mismo tiempo, de HGF (100 ng/ml) y los anticuerpos a probar (10 µg/ml). Después de 72 horas de incubación, se fijaron y tiñeron colonias con violeta cristal al 0.2% en metanol y estimaron por dispersión visualmente. Cada condición probada se probó en triplicado y se realizaron 3 experimentos independientes.
La adición de HGF a células SK-HEP-1 induce una dispersión significante de células (Figuras 10A y 10B). El anticuerpo 9G4 introducido como un control de isotipo no tiene efecto ni solo o en la presencia de HGF. Como se espera, el anticuerpo 5D5 exhibe un efecto agonista significante solo y no se observa efecto inhibitorio cuando se agrega 5D5 con HGF (Figura 10A). No se observa efecto agonista ni con 11E1 (Figura 10A) ni con 224G11 (Figura 10B) agregados solos. Un efecto inhibitorio muy significante de estos anticuerpos se demuestra en la presencia de HGF (Figuras 10A y 10B).
Ejemplo 8: Ensayo de Tubulogénesis Tri-dimensional
Células SK-HEP-1 se sembraron a 1.104 células/pozo en una placa de 24 pocillos en DMEM con FCS al 10%/Matrigel (50/50) e incubaron por 30 minutos antes de adición, al mismo tiempo, de HGF (100 ng/ml) y los anticuerpos a probar (10 µg/ml). Después de 7 días de incubación, las células se estimaron visualmente por formación de tubos. Cada condición probada se realizó como un triplicado y se efectuaron 3 experimentos independientes.
La adición de HGF induce una formación de tubo SK-HEP-1 significante (Figura 11). El anticuerpo 9G4 introducido como un control de isotipo no tuvo efecto ni solo ni en la presencia de HGF. Como se espera, el anticuerpo 5D5 exhibe un efecto agonista significante solo y no se observa efecto inhibitorio cuando 5D5 se agrega con HGF. No se observa efecto agonista con 11E1, 223C4 y 224G11 agregados solos y un efecto inhibitorio completo se demuestra tanto con 11E1 como 223C4, en la presencia de HGF. Una inhibición parcial pero significativa se observa con 224G11Mab.
Ejemplo 9: Formación de Esferoide
Para evaluar la capacidad de anticuerpos anti-c-Met para inhibir el crecimiento de tumor in vitro, en un modelo cercano a una situación in vivo, se generaron esferoides de células de glioblastoma humano U-87MG (ATCC # HTB14). Las células desarrolladas como una monocapa se desprendieron con tripsina-EDTA y resuspenden en medio de cultivo celular completo (DMEM) suplementado con FBS al 10%. Se iniciaron esferoides al inocular 625 células en pocillos sencillos de fondo redondo, 96 placas en DMEM-FCS al 10%. Para impedir adhesión celular a un sustrato, las placas se prerrevistieron con polyHEMA en etanol al 95% y secaron al aire a temperatura ambiente. Las placas se incubaron bajo condiciones de cultivo celular estándar a 37 grados C, CO2 al 5% en incubadoras humidificadas.
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Applications Claiming Priority (7)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US92978907P | 2007-07-12 | 2007-07-12 | |
| EP07301231 | 2007-07-12 | ||
| EP07301231A EP2014681A1 (en) | 2007-07-12 | 2007-07-12 | Novel antibodies inhibiting c-met dimerization, and uses thereof |
| US929789P | 2007-07-12 | ||
| US2063908P | 2008-01-11 | 2008-01-11 | |
| US20639P | 2008-01-11 | ||
| PCT/EP2008/059026 WO2009007427A2 (en) | 2007-07-12 | 2008-07-10 | Novel antibodies inhibiting c-met dimerization, and uses thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| ES2542059T3 true ES2542059T3 (es) | 2015-07-30 |
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ID=38871713
Family Applications (5)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| ES11162718.8T Active ES2686335T3 (es) | 2007-07-12 | 2008-07-10 | Nuevos anticuerpos que inhiben la dimerización de c-Met y sus utilizaciones |
| ES12183761.1T Active ES2686693T3 (es) | 2007-07-12 | 2008-07-10 | Nuevos anticuerpos que inhiben la dimerización de c-Met y sus utilizaciones |
| ES11162720.4T Active ES2678495T3 (es) | 2007-07-12 | 2008-07-10 | Nuevos anticuerpos que inhiben la dimerización de c-Met y sus utilizaciones |
| ES12183763.7T Active ES2693542T3 (es) | 2007-07-12 | 2008-07-10 | Nuevos anticuerpos que inhiben la dimerización de c-Met y sus utilizaciones |
| ES08786050.8T Active ES2542059T3 (es) | 2007-07-12 | 2008-07-10 | Nuevos anticuerpos que inhiben la dimerización de c-Met y sus utilizaciones |
Family Applications Before (4)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| ES11162718.8T Active ES2686335T3 (es) | 2007-07-12 | 2008-07-10 | Nuevos anticuerpos que inhiben la dimerización de c-Met y sus utilizaciones |
| ES12183761.1T Active ES2686693T3 (es) | 2007-07-12 | 2008-07-10 | Nuevos anticuerpos que inhiben la dimerización de c-Met y sus utilizaciones |
| ES11162720.4T Active ES2678495T3 (es) | 2007-07-12 | 2008-07-10 | Nuevos anticuerpos que inhiben la dimerización de c-Met y sus utilizaciones |
| ES12183763.7T Active ES2693542T3 (es) | 2007-07-12 | 2008-07-10 | Nuevos anticuerpos que inhiben la dimerización de c-Met y sus utilizaciones |
Country Status (36)
| Country | Link |
|---|---|
| US (6) | US8329173B2 (es) |
| EP (6) | EP2014681A1 (es) |
| JP (4) | JP5562844B2 (es) |
| KR (4) | KR101701686B1 (es) |
| CN (4) | CN103183739B (es) |
| AR (4) | AR067517A1 (es) |
| AU (1) | AU2008274171B2 (es) |
| BR (1) | BRPI0815564B8 (es) |
| CA (4) | CA2888691C (es) |
| CL (1) | CL2008002015A1 (es) |
| CO (1) | CO6170364A2 (es) |
| CY (1) | CY1116608T1 (es) |
| DK (1) | DK2188312T3 (es) |
| ES (5) | ES2686335T3 (es) |
| HK (3) | HK1149024A1 (es) |
| HR (1) | HRP20150887T1 (es) |
| HU (1) | HUE026026T2 (es) |
| IL (5) | IL203114A (es) |
| MA (1) | MA31977B1 (es) |
| ME (1) | ME02341B (es) |
| MX (3) | MX353449B (es) |
| MY (1) | MY158756A (es) |
| NZ (4) | NZ613716A (es) |
| PA (1) | PA8789201A1 (es) |
| PH (3) | PH12018502039A1 (es) |
| PL (1) | PL2188312T3 (es) |
| PT (1) | PT2188312E (es) |
| RS (1) | RS54197B1 (es) |
| RU (1) | RU2552161C2 (es) |
| SA (1) | SA08290423B1 (es) |
| SG (1) | SG183015A1 (es) |
| SI (1) | SI2188312T1 (es) |
| TN (1) | TN2010000016A1 (es) |
| TW (4) | TWI549965B (es) |
| WO (1) | WO2009007427A2 (es) |
| ZA (1) | ZA201000966B (es) |
Families Citing this family (99)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2143441A1 (en) * | 2008-07-08 | 2010-01-13 | Pierre Fabre Medicament | Combination of a c-Met antagonist and an aminoheteroaryl compound for the treatment of cancer |
| PA8849001A1 (es) | 2008-11-21 | 2010-06-28 | Lilly Co Eli | Anticuerpos de c-met |
| US20140112911A9 (en) * | 2008-12-02 | 2014-04-24 | Liliane Goetsch | Novel anti-cmet antibody |
| US9469691B2 (en) * | 2008-12-02 | 2016-10-18 | Pierre Fabre Medicament | Anti-cMET antibody |
| US8545839B2 (en) * | 2008-12-02 | 2013-10-01 | Pierre Fabre Medicament | Anti-c-Met antibody |
| AR074439A1 (es) * | 2008-12-02 | 2011-01-19 | Pf Medicament | Anticuerpo anti-cmet (receptor c-met) |
| WO2010115553A1 (en) | 2009-04-07 | 2010-10-14 | Roche Glycart Ag | Bispecific anti-erbb-2/anti-c-met antibodies |
| KR20110124369A (ko) | 2009-04-07 | 2011-11-16 | 로슈 글리카트 아게 | 이중특이적 항ｅｒｂｂ3／항ｃｍｅｔ 항체 |
| EP2287197A1 (en) | 2009-08-21 | 2011-02-23 | Pierre Fabre Medicament | Anti-cMET antibody and its use for the detection and the diagnosis of cancer |
| KR101671378B1 (ko) * | 2009-10-30 | 2016-11-01 | 삼성전자 주식회사 | ｃ-Ｍｅｔ에 특이적으로 결합하는 항체 및 그의 용도 |
| KR101748707B1 (ko) | 2009-11-27 | 2017-06-20 | 삼성전자주식회사 | ｃ-Ｍｅｔ에 특이적으로 결합하는 항체 및 그를 이용한 암 진단용 키트 |
| SG10201501767VA (en) * | 2010-03-10 | 2015-05-28 | Genmab As | Monoclonal antibodies against c-met |
| EP2576622A4 (en) * | 2010-06-01 | 2013-11-27 | Univ Monash | ANTIBODIES AGAINST RECEPTOR TYROSINE KINASE C-MET |
| EP2402370A1 (en) * | 2010-06-29 | 2012-01-04 | Pierre Fabre Médicament | Novel antibody for the diagnosis and/or prognosis of cancer |
| US20130315895A1 (en) | 2010-07-01 | 2013-11-28 | Takeda Pharmaceutical Company Limited | COMBINATION OF A cMET INHIBITOR AND AN ANTIBODY TO HGF AND/OR cMET |
| SG187886A1 (en) | 2010-08-31 | 2013-04-30 | Genentech Inc | Biomarkers and methods of treatment |
| KR20140019284A (ko) * | 2010-09-03 | 2014-02-14 | 아카데미아 시니카 | 항-C-Met 항체 및 이의 이용 방법들 |
| US11644471B2 (en) | 2010-09-30 | 2023-05-09 | Ablynx N.V. | Techniques for predicting, detecting and reducing aspecific protein interference in assays involving immunoglobulin single variable domains |
| WO2012042026A1 (en) | 2010-09-30 | 2012-04-05 | Ablynx Nv | Biological materials related to c-met |
| EP2635602B1 (en) | 2010-11-03 | 2016-09-07 | Argen-X Nv | Anti c-met antibodies |
| US20130236467A1 (en) | 2010-11-24 | 2013-09-12 | Jeremy Griggs | Multispecific antigen binding proteins targeting hgf |
| KR101444837B1 (ko) | 2011-06-03 | 2014-09-30 | 한국생명공학연구원 | ＨＧＦ 활성을 가지는 ｃ-Ｍｅｔ에 대한 인간항체 및 이의 용도 |
| EP3466972A1 (en) | 2011-06-23 | 2019-04-10 | Ablynx NV | Serum albumin binding proteins |
| HUE031828T2 (en) | 2011-06-23 | 2017-08-28 | Ablynx Nv | Procedures for Predicting, Detecting, and Reducing Aspiration Protein Interference in Assays Containing Immunoglobulin Variable Single Domain |
| KR20140045440A (ko) * | 2011-06-30 | 2014-04-16 | 제넨테크, 인크. | 항-c-met 항체 제제 |
| JP2015527869A (ja) | 2011-08-26 | 2015-09-24 | メリマック ファーマシューティカルズ インコーポレーティッド | タンデムＦｃ二重特異性抗体 |
| AU2012312515A1 (en) | 2011-09-19 | 2014-03-13 | Genentech, Inc. | Combination treatments comprising c-met antagonists and B-raf antagonists |
| US9201074B2 (en) | 2011-09-20 | 2015-12-01 | Eli Lilly And Company | Anti-c-Met antibodies |
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