The Experts below are selected from a list of 360 Experts worldwide ranked by ideXlab platform

Jacek Otlewski - One of the best experts on this subject based on the ideXlab platform.

  • purification and in vitro Evaluation of an anti hEr2 affibody MonomEthyl Auristatin E conjugatE in hEr2 positivE cancEr cElls
    Biology, 2021
    Co-Authors: Isabella Damiani, Jacek Otlewski, Silvia Castiglioni, Alicja M Sochajgregorczyk, Fabrizia Bonacina, Irma Colombo, Valentina Rusconi, Alberto Corsini, Stefano Bellosta
    Abstract:

    A promising approach for thE dEvElopmEnt of high-affinity tumor targEting ADCs is thE usE of EnginEErEd protEin drugs, such as affibody molEculEs, which rEprEsEnt a valuablE altErnativE to monoclonal antibodiEs (mAbs) in cancEr-targEtEd thErapy. WE dEvElopEd a mEthod for a morE EfficiEnt purification of thE ZHER2:2891DCS affibody conjugatEd with thE cytotoxic antimitotic agEnt Auristatin E (MMAE), and its Efficacy was tEstEd in vitro on cEll viability, prolifEration, migration, and apoptosis. ThE EffEcts of ZHER2:2891DCS-MMAE wErE comparEd with thE clinically approvEd monoclonal antibody trastuzumab (HErcEptin®). To dEmonstratE that ZHER2:2891DCS-MMAE can sElEctivEly targEt HER2 ovErExprEssing tumor cElls, wE usEd thrEE diffErEnt cEll linEs: thE human adEnocarcinoma cEll linEs SK-BR-3 and ZR-75-1, both ovErExprEssing HER2, and thE triplE-nEgativE brEast cancEr cEll linE MDA-MB-231. MTT assay showEd that ZHER2:2891DCS-MMAE inducEs a significant timE-dEpEndEnt toxic EffEct in SK-BR-3 cElls. A 30% rEduction of cEll viability was alrEady found aftEr 10 min ExposurE at a concEntration of 7 nM (IC50 of 80.2 nM). On thE contrary, MDA-MB-231 cElls, which ExprEss basal lEvEls of HER2, wErE not affEctEd by thE conjugatE. ThE cytotoxic EffEct of thE ZHER2:2891DCS-MMAE was confirmEd by mEasuring apoptosis by flow cytomEtry. In SK-BR-3 cElls, incrEasing concEntrations of conjugatEd affibody inducEd cEll dEath starting from 10 min of trEatmEnt, with thE strongEst EffEct obsErvEd aftEr 48 h. OvErall, thEsE rEsults dEmonstratE that thE ADC, formEd by thE anti-HER2 affibody conjugatEd to MonomEthyl Auristatin E, EfficiEntly intEracts with high affinity with HER2 positivE cancEr cElls in vitro, allowing thE sElEctivE and spEcific dElivEry of thE cytotoxic payload.

  • FGF2 Dual WarhEad ConjugatE with MonomEthyl Auristatin E and α-Amanitin Displays a Cytotoxic EffEct towards CancEr CElls OvErproducing FGF REcEptor 1
    International journal of molecular sciences, 2018
    Co-Authors: Karolina W. Świderska, Anna Szlachcic, Malgorzata Zakrzewska, Łukasz Opaliński, Jacek Otlewski
    Abstract:

    In thE rapidly dEvEloping fiEld of targEtEd cancEr thErapy thErE is growing intErEst towards thErapEutics combining two or morE compounds to achiEvE synErgistic action and minimizE thE chancE of cancEr rEsistancE to trEatmEnt. WE dEvElopEd a fibroblast growth factor 2 (FGF2)-conjugatE bEaring two cytotoxic drugs with indEpEndEnt modE of action: α-amanitin and MonomEthyl Auristatin E. Drugs arE covalEntly attachEd to thE targEting protEin in a sitE-spEcific mannEr via malEimidE-thiol conjugation and Cu(I)-catalyzEd alkynE-azidE cycloaddition. ThE dual warhEad conjugatE binds to FGF rEcEptor 1 (FGFR1) and utilizEs rEcEptor-mEdiatEd Endocytosis for sElEctivE intErnalization into cancEr cElls with FGFR1. ThE dEvElopEd conjugatE displays high cytotoxicity towards all tEstEd FGFR1-positivE cEll linEs. Most importantly, thE improvEd cytotoxic EffEct of both drugs is obsErvEd for lung cancEr cEll linE NCI-H446. ThE singlE drug-FGF2 conjugatEs havE no impact on thE viability of NCI-H446 cElls, whErEas thE dual warhEad-FGF2 conjugatE sElEctivEly and EfficiEntly kills thEsE FGFR1 positivE cancEr cElls. DuE to thE divErsifiEd modE of action thE dual warhEad-FGF2 conjugatE may ovErcomE thE potEntial acquirEd rEsistancE of FGFR1-ovErproducing cancEr cElls towards singlE cytotoxic drugs.

  • Cytotoxic ConjugatEs of Fibroblast Growth Factor 2 (FGF2) with MonomEthyl Auristatin E for EffEctivE Killing of CElls ExprEssing FGF REcEptors.
    ACS omega, 2017
    Co-Authors: Mateusz Adam Krzyscik, Malgorzata Zakrzewska, Vigdis Sørensen, Aleksandra Sokolowska-wedzina, Michal Lobocki, Karolina Weronika Swiderska, Daniel Krowarsch, Antoni G Wiedlocha, Jacek Otlewski
    Abstract:

    Antibody–drug conjugatEs (ADCs) arE a nEw class of anticancEr thErapEutics that combinE thE sElEctivity of targEtEd trEatmEnt, EnsurEd by monoclonal antibodiEs, with thE potEncy of thE cytotoxic agEnt. HErE, wE appliEd an analogous approach, but instEad of an antibody, wE usEd fibroblast growth factor 2 (FGF2). FGF2 is a natural ligand of fibroblast growth factor rEcEptor 1 (FGFR1), a cEll-surfacE rEcEptor rEportEd to bE ovErExprEssEd in sEvEral typEs of tumors. WE dEvElopEd and charactErizEd FGF2 conjugatEs containing a dEfinEd numbEr of molEculEs of highly cytotoxic drug MonomEthyl Auristatin E (MMAE). ThEsE conjugatEs EffEctivEly targEtEd FGFR1-ExprEssing cElls, wErE intErnalizEd upon FGFR1-mEdiatEd Endocytosis, and, in consEquEncE, rEvEalEd high cytotoxicity, which was clEarly rElatEd to thE FGFR1 ExprEssion lEvEl. Among thE conjugatEs tEstEd, thE most potEnt was that bEaring thrEE MMAE molEculEs, showing that thE cytotoxicity of protEin–drug conjugatEs in vitro is dirEctly dEpEndEnt on drug loading.

  • Cytotoxic ConjugatEs of Fibroblast Growth Factor 2 (FGF2) with MonomEthyl Auristatin E for EffEctivE Killing of CElls ExprEssing FGF REcEptors
    2017
    Co-Authors: Mateusz Adam Krzyscik, Malgorzata Zakrzewska, Vigdis Sørensen, Aleksandra Sokolowska-wedzina, Michal Lobocki, Karolina Weronika Swiderska, Daniel Krowarsch, Antoni G Wiedlocha, Jacek Otlewski
    Abstract:

    Antibody–drug conjugatEs (ADCs) arE a nEw class of anticancEr thErapEutics that combinE thE sElEctivity of targEtEd trEatmEnt, EnsurEd by monoclonal antibodiEs, with thE potEncy of thE cytotoxic agEnt. HErE, wE appliEd an analogous approach, but instEad of an antibody, wE usEd fibroblast growth factor 2 (FGF2). FGF2 is a natural ligand of fibroblast growth factor rEcEptor 1 (FGFR1), a cEll-surfacE rEcEptor rEportEd to bE ovErExprEssEd in sEvEral typEs of tumors. WE dEvElopEd and charactErizEd FGF2 conjugatEs containing a dEfinEd numbEr of molEculEs of highly cytotoxic drug MonomEthyl Auristatin E (MMAE). ThEsE conjugatEs EffEctivEly targEtEd FGFR1-ExprEssing cElls, wErE intErnalizEd upon FGFR1-mEdiatEd Endocytosis, and, in consEquEncE, rEvEalEd high cytotoxicity, which was clEarly rElatEd to thE FGFR1 ExprEssion lEvEl. Among thE conjugatEs tEstEd, thE most potEnt was that bEaring thrEE MMAE molEculEs, showing that thE cytotoxicity of protEin–drug conjugatEs in vitro is dirEctly dEpEndEnt on drug loading

Filip S. Ekholm - One of the best experts on this subject based on the ideXlab platform.

  • incrEasing thE potEntial of thE Auristatin cancEr drug family by shifting thE conformational Equilibrium
    Molecular Pharmaceutics, 2019
    Co-Authors: Iris Katariina Sokka, Filip S. Ekholm, Mikael P. Johansson
    Abstract:

    MonomEthyl Auristatin E and MonomEthyl Auristatin F arE widEly usEd cytotoxic agEnts in antibody–drug conjugatEs (ADCs), a group of promising cancEr drugs. ThE ADCs spEcifically targEt cancEr cElls, rElEasing thE Auristatins insidE, which rEsults in thE prEvEntion of mitosis. ThE Auristatins suffEr from a potEntially sErious flaw, howEvEr. In solution, thE molEculEs Exist in an Equal mixturE of two conformErs, cis and trans. Only thE trans-isomEr is biologically activE and thE isomErization procEss, i.E., thE convErsion of cis to trans is slow. This significantly diminishEs thE EfficiEncy of thE drugs and thEir corrEsponding ADCs, and pErhaps morE importantly, raisEs concErns ovEr drug safEty. ThE potEncy of thE Auristatins would bE EnhancEd by dEcrEasing thE amount of thE biologically inactivE isomEr, EithEr by stabilizing thE trans-isomEr or dEstabilizing thE cis-isomEr. HErE, wE follow thE computEr-aidEd dEsign stratEgy of shifting thE conformational Equilibrium and Employ high-lEvEl quantum chEmical m...

  • incrEasing thE potEntial of thE Auristatin cancEr drug family by shifting thE conformational Equilibrium
    Molecular Pharmaceutics, 2019
    Co-Authors: Iris Katariina Sokka, Filip S. Ekholm, Mikael P. Johansson
    Abstract:

    MonomEthyl Auristatin E and MonomEthyl Auristatin F arE widEly usEd cytotoxic agEnts in antibody-drug conjugatEs (ADCs), a group of promising cancEr drugs. ThE ADCs spEcifically targEt cancEr cElls, rElEasing thE Auristatins insidE, which rEsults in thE prEvEntion of mitosis. ThE Auristatins suffEr from a potEntially sErious flaw, howEvEr. In solution, thE molEculEs Exist in an Equal mixturE of two conformErs, cis and trans. Only thE trans-isomEr is biologically activE and thE isomErization procEss, i.E., thE convErsion of cis to trans is slow. This significantly diminishEs thE EfficiEncy of thE drugs and thEir corrEsponding ADCs, and pErhaps morE importantly, raisEs concErns ovEr drug safEty. ThE potEncy of thE Auristatins would bE EnhancEd by dEcrEasing thE amount of thE biologically inactivE isomEr, EithEr by stabilizing thE trans-isomEr or dEstabilizing thE cis-isomEr. HErE, wE follow thE computEr-aidEd dEsign stratEgy of shifting thE conformational Equilibrium and Employ high-lEvEl quantum chEmical modEling to idEntify promising candidatEs for improvEd Auristatins. CouplEd clustEr calculations prEdict that a simplE halogEnation in thE norEphEdrinE/phEnylalaninE rEsiduEs shifts thE isomEr Equilibrium almost complEtEly toward thE activE trans-conformation, duE to EnhancEd intramolEcular intEractions spEcific to thE activE isomEr.

  • Hydrophilic MonomEthyl Auristatin E DErivativEs as NovEl CandidatEs for thE DEsign of Antibody-Drug ConjugatEs
    MDPI AG, 2018
    Co-Authors: Filip S. Ekholm, Suvi-katriina Ruokonen, Marina Redón, Virve Pitkänen, Anja Vilkman, Juhani Saarinen, Jari Helin, Tero Satomaa, Susanne K. Wiedmer
    Abstract:

    Antibody-drug conjugatEs (ADCs) arE promising statE-of-thE-art biopharmacEutical drugs for sElEctivE drug-dElivEry applications and thE trEatmEnt of disEasEs such as cancEr. ThE idEa bEhind thE ADC tEchnology is rEmarkablE as it combinEs thE highly sElEctivE targEting capacity of monoclonal antibodiEs with thE cancEr-killing ability of potEnt cytotoxic agEnts. ThE continuous dEvElopmEnt of improvEd ADCs rEquirEs systEmatic studiEs on thE naturE and EffEcts of warhEad modification. REcEntly, wE focusEd on thE hydrophilic modification of MonomEthyl Auristatin E (MMAE), thE most widEly usEd cytotoxic agEnt in currEnt clinical trial ADCs. HErEin, wE rEport on thE usE of micEllar ElEctrokinEtic chromatography (MEKC) for studying thE hydrophobic charactEr of modifiEd MMAE dErivativEs. Our data rEvEal a connEction bEtwEEn thE hydrophobicity of thE modifiEd warhEads as frEE molEculEs and thEir cytotoxic activity. In addition, MMAE-trastuzumab ADCs wErE constructEd and EvaluatEd in prEliminary cytotoxic assays

  • NEw insight on thE structural fEaturEs of thE cytotoxic Auristatins MMAE and MMAF rEvEalEd by combinEd NMR spEctroscopy and quantum chEmical modElling.
    Scientific reports, 2017
    Co-Authors: Mikael P. Johansson, Hannu Maaheimo, Filip S. Ekholm
    Abstract:

    Antibody-drug conjugatEs (ADCs) arE EmErging as a promising class of sElEctivE drug dElivEry systEms in thE battlE against cancEr and othEr disEasEs. ThE Auristatins MonomEthyl Auristatin E (MMAE) and MonomEthyl Auristatin F (MMAF) appEar as thE cytotoxic drug in almost half of thE statE-of-thE-art ADCs on thE markEt or in latE stagE clinical trials. HErE, wE prEsEnt thE first complEtE NMR spEctroscopic charactErisation of thEsE challEnging molEculEs, and invEstigatE thEir structural propErtiEs by a combinEd NMR and quantum chEmical modElling approach. WE find that in solution, half of thE drug molEculEs arE lockEd in an inactivE conformation, sEvErEly dEcrEasing thEir EfficiEncy, and potEntially incrEasing thE risk of sidE-EffEcts. FurthErmorE, wE idEntify sitEs suscEptiblE to futurE modification, in ordEr to potEntially improvE thE pErformancE of thEsE drugs.

  • introducing glycolinkErs for thE functionalization of cytotoxic drugs and applications in antibody drug conjugation chEmistry
    ChemMedChem, 2016
    Co-Authors: Filip S. Ekholm, Virve Pitkänen, Anja Vilkman, Juhani Saarinen, Jari Helin, Henna Pynnönen, Tero Satomaa
    Abstract:

    Antibody–drug conjugatEs (ADCs) arE promising altErnativEs to nakEd antibodiEs for sElEctivE drug-dElivEry applications and trEatmEnt of disEasEs such as cancEr. Construction of ADCs rEliEs upon sitE-sElEctivE, EfficiEnt and mild conjugation tEchnologiEs. ThE choicE of a chEmical linkEr is EspEcially important, as it affEcts thE ovErall propErtiEs of thE ADC. WE EnvisionEd that hydrophilic bifunctional chEmical linkErs basEd on carbohydratEs would bE a usEful class of dErivatization agEnts for thE construction of linkEr–drug conjugatEs and ADCs. HErEin wE dEscribE thE synthEsis of carbohydratE-basEd dErivatization agEnts, glycolinkEr–drug conjugatEs fEaturing thE tubulin inhibitor MonomEthyl Auristatin E and an ADC basEd on an anti-EGFR antibody. In addition, an initial in vitro cytotoxicity Evaluation of thE individual componEnts and thE ADC is providEd against EGFR-positivE cancEr cElls.

Tero Satomaa - One of the best experts on this subject based on the ideXlab platform.

  • Hydrophilic MonomEthyl Auristatin E DErivativEs as NovEl CandidatEs for thE DEsign of Antibody-Drug ConjugatEs
    MDPI AG, 2018
    Co-Authors: Filip S. Ekholm, Suvi-katriina Ruokonen, Marina Redón, Virve Pitkänen, Anja Vilkman, Juhani Saarinen, Jari Helin, Tero Satomaa, Susanne K. Wiedmer
    Abstract:

    Antibody-drug conjugatEs (ADCs) arE promising statE-of-thE-art biopharmacEutical drugs for sElEctivE drug-dElivEry applications and thE trEatmEnt of disEasEs such as cancEr. ThE idEa bEhind thE ADC tEchnology is rEmarkablE as it combinEs thE highly sElEctivE targEting capacity of monoclonal antibodiEs with thE cancEr-killing ability of potEnt cytotoxic agEnts. ThE continuous dEvElopmEnt of improvEd ADCs rEquirEs systEmatic studiEs on thE naturE and EffEcts of warhEad modification. REcEntly, wE focusEd on thE hydrophilic modification of MonomEthyl Auristatin E (MMAE), thE most widEly usEd cytotoxic agEnt in currEnt clinical trial ADCs. HErEin, wE rEport on thE usE of micEllar ElEctrokinEtic chromatography (MEKC) for studying thE hydrophobic charactEr of modifiEd MMAE dErivativEs. Our data rEvEal a connEction bEtwEEn thE hydrophobicity of thE modifiEd warhEads as frEE molEculEs and thEir cytotoxic activity. In addition, MMAE-trastuzumab ADCs wErE constructEd and EvaluatEd in prEliminary cytotoxic assays

  • introducing glycolinkErs for thE functionalization of cytotoxic drugs and applications in antibody drug conjugation chEmistry
    ChemMedChem, 2016
    Co-Authors: Filip S. Ekholm, Virve Pitkänen, Anja Vilkman, Juhani Saarinen, Jari Helin, Henna Pynnönen, Tero Satomaa
    Abstract:

    Antibody–drug conjugatEs (ADCs) arE promising altErnativEs to nakEd antibodiEs for sElEctivE drug-dElivEry applications and trEatmEnt of disEasEs such as cancEr. Construction of ADCs rEliEs upon sitE-sElEctivE, EfficiEnt and mild conjugation tEchnologiEs. ThE choicE of a chEmical linkEr is EspEcially important, as it affEcts thE ovErall propErtiEs of thE ADC. WE EnvisionEd that hydrophilic bifunctional chEmical linkErs basEd on carbohydratEs would bE a usEful class of dErivatization agEnts for thE construction of linkEr–drug conjugatEs and ADCs. HErEin wE dEscribE thE synthEsis of carbohydratE-basEd dErivatization agEnts, glycolinkEr–drug conjugatEs fEaturing thE tubulin inhibitor MonomEthyl Auristatin E and an ADC basEd on an anti-EGFR antibody. In addition, an initial in vitro cytotoxicity Evaluation of thE individual componEnts and thE ADC is providEd against EGFR-positivE cancEr cElls.

Stephen C Alley - One of the best experts on this subject based on the ideXlab platform.

  • cyp3a mEdiatEd drug drug intEraction potEntial and ExcrEtion of brEntuximab vEdotin an antibody drug conjugatE in patiEnts with cd30 positivE hEmatologic malignanciEs
    The Journal of Clinical Pharmacology, 2013
    Co-Authors: Tae H Han, Jeffrey Matous, Ajay K Gopal, Radhakrishnan Ramchandren, Andre Goy, Robert T Chen, Maureen Cooper, Laurie E Grove, Stephen C Alley, Carmel M Lynch
    Abstract:

    BrEntuximab vEdotin is an antibody-drug conjugatE (ADC) that sElEctivEly dElivErs MonomEthyl Auristatin E (MMAE) into CD30-ExprEssing cElls. This study EvaluatEd thE CYP3A-mEdiatEd drug-drug intEraction potEntial of brEntuximab vEdotin and thE ExcrEtion of MMAE. Two 21-day cyclEs of brEntuximab vEdotin (1.2 or 1.8 mg/kg intravEnously) wErE administErEd to 56 patiEnts with CD30-positivE hEmatologic malignanciEs. Each patiEnt also rEcEivEd EithEr a sEnsitivE CYP3A substratE (midazolam), an EffEctivE inducEr (rifampin), or a strong inhibitor (kEtoconazolE). BrEntuximab vEdotin did not affEct midazolam ExposurEs. ADC ExposurEs wErE unaffEctEd by concomitant rifampin or kEtoconazolE; howEvEr, MMAE ExposurEs wErE lowEr with rifampin and highEr with kEtoconazolE. ThE short-tErm safEty profilE of brEntuximab vEdotin in this study was gEnErally consistEnt with historic clinical obsErvations. ThE most common advErsE EvEnts wErE nausEa, fatiguE, diarrhEa, hEadachE, pyrExia, and nEutropEnia. OvEr a 1-wEEk pEriod, ∼23.5% of intact MMAE was rEcovErEd aftEr administration of brEntuximab vEdotin; all othEr spEciEs wErE bElow thE limit of quantitation. ThE primary ExcrEtion routE is via fEcEs (mEdian 72% of thE rEcovErEd MMAE). ThEsE rEsults suggEst that brEntuximab vEdotin (1.8 mg/kg) and MMAE arE nEithEr inhibitors nor inducErs of CYP3A; howEvEr, MMAE is a substratE of CYP3A.

  • potEnt antitumor activity of thE anti cd19 Auristatin antibody drug conjugatE hbu12 vcmmaE against rituximab sEnsitivE and rEsistant lymphomas
    Blood, 2009
    Co-Authors: Hans-peter Gerber, Stephen C Alley, May Kungsutherland, Ivan Stone, Caroll Morristilden, Jamie Miyamoto, Renee Mccormick, Nicole M Okeley, Brad Hayes, Francisco J Hernandezilizaliturri
    Abstract:

    DEspitE major advancEs in thE trEatmEnt of non-Hodgkin lymphoma (NHL), including thE usE of chEmothErapEutic agEnts and thE anti-CD20 antibody rituximab, thE majority of patiEnts EvEntually rElapsE, and salvagE trEatmEnts with non–cross-rEsistant compounds arE nEEdEd to furthEr improvE patiEnt survival. HErE, wE EvaluatEd thE antitumor EffEcts of thE microtubulE dEstabilizing agEnt MonomEthyl Auristatin E (MMAE) conjugatEd to thE humanizEd anti-CD19 antibody hBU12 via a protEasE-sEnsitivE valinE-citrullinE (vc) dipEptidE linkEr. hBU12-vcMMAE inducEd potEnt tumor cEll killing against rituximab-sEnsitivE and -rEsistant NHL cEll linEs. CD19 can form hEtErodimErs with CD21, and high lEvEls of CD21 wErE rEportEd to intErfErE nEgativEly with thE activity of CD19-targEtEd thErapEutics. HowEvEr, wE obsErvEd comparablE intErnalization, intracEllular trafficking, and drug rElEasE in CD21low and CD21high, rituximab-sEnsitivE and -rEfractory lymphomas trEatEd with hBU12-vcMMAE. FurthErmorE, high ratEs of durablE rEgrEssions in micE implantEd with thEsE tumors wErE obsErvEd, suggEsting that both rituximab rEsistancE and CD21 ExprEssion lEvEls do not impact on thE activity of hBU12-vcMMAE. CombinEd, our data suggEst that hBU12-vcMMAE may rEprEsEnt a promising addition to thE trEatmEnt options for rituximab rEfractory NHL and othEr hEmatologic malignanciEs, including acutE lymphoblastic lEukEmia.

  • rEduction alkylation stratEgiEs for thE modification of spEcific monoclonal antibody disulfidEs
    Bioconjugate Chemistry, 2005
    Co-Authors: Kevin S Beam, Charles G Cerveny, Peter D Senter, Kevin J Hamblett, Richard S Blackmore, Michael Torgov, Felicia G M Handley, Nathan Ihle, Stephen C Alley
    Abstract:

    SitE-spEcific conjugation of small molEculEs and EnzymEs to monoclonal antibodiEs has broad utility in thE formation of conjugatEs for thErapEutic, diagnostic, or structural applications. PrEcisE control ovEr thE location of conjugation would yiEld highly homogEnEous matErials that could havE improvEd biological propErtiEs. WE dEscribE for thE first timE chEmical rEduction and oxidation mEthods that lEad to prEfErEntial clEavagE of particular monoclonal antibody intErchain disulfidEs using thE anti-CD30 IgG1 monoclonal antibody cAC10. Alkylation of thE rEsulting cAC10 cystEinE thiols with thE potEnt antimitotic agEnt MonomEthyl Auristatin E (MMAE) EnablEd thE assignmEnt of drug conjugation location by purification with hydrophobic intEraction chromatography followEd by analysis using rEvErsEd-phasE HPLC and capillary ElEctrophorEsis. ThEsE analytical mEthods dEmonstratEd that trEating cAC10 with rEducing agEnts such as DTT causEd prEfErEntial rEduction of hEavy-light chain disulfidEs, whilE rEoxidation of...

Yujie Zhang - One of the best experts on this subject based on the ideXlab platform.

  • Activating Autophagy EnhancEd thE Antitumor EffEct of Antibody Drug ConjugatEs Rituximab-MonomEthyl Auristatin E
    Frontiers in immunology, 2018
    Co-Authors: Yichen Wang, Xuyao Zhang, Jiajun Fan, Wei Chen, Jingyun Luan, Yanyang Nan, Shaofei Wang, Qicheng Chen, Yujie Zhang
    Abstract:

    Antibody drug conjugatE (ADC) showEd potEnt thErapEutic Efficacy in sEvEral typEs of cancErs. ThE rolE of autophagy in anti-tumor EffEcts of ADC rEmains unclEar. In this study, a novEl ADC, Rituximab-MonomEthyl Auristatin E (MMAE) with a Val-Cit clEavablE linkEr, was dEsignEd to invEstigatE its thErapEutic Efficacy against Non-Hodgkin Lymphoma (NHL) as wEll as thE undErlying mEchanisms. Our rEsults showEd that Rituximab-MMAE ElicitEd caspasE-3-dEpEndEnt apoptosis in NHL cElls and ExhibitEd potEnt thErapEutic Efficacy in vivo. Autophagy, which was charactErizEd by up-rEgulatEd LC3-II ExprEssion, and formation and accumulation of autophagosomEs, was triggErEd during thE ADC trEatmEnt. MEanwhilE, inactivation of Akt/mTOR pathway was shown to bE involvEd in thE autophagy triggErEd by Rituximab-MMAE, indicating a probablE mEchanism of ADC-initiatEd autophagy. Importantly, inhibition of autophagy by chloroquinE supprEssEd thE ADC-inducEd apoptosis, whilE activating autophagy by rapamycin significantly EnhancEd thE thErapEutic EffEct of Rituximab-MMAE both in vitro and in vivo. Our data ElucidatEd thE critical rElationship bEtwEEn autophagy and apoptosis in Rituximab-MMAE-basEd thErapy and highlightEd thE potEntial approach for NHL thErapy by combinEd administration of ADC and autophagy activation.

  • Activating Autophagy EnhancEd thE Antitumor EffEct of Antibody Drug ConjugatEs Rituximab-MonomEthyl Auristatin E
    Frontiers Media S.A., 2018
    Co-Authors: Yichen Wang, Xuyao Zhang, Jiajun Fan, Wei Chen, Jingyun Luan, Yanyang Nan, Shaofei Wang, Qicheng Chen, Yujie Zhang
    Abstract:

    BackgroundAntibody drug conjugatE (ADC) showEd potEnt thErapEutic Efficacy in sEvEral typEs of cancErs. ThE rolE of autophagy in antitumor EffEcts of ADC rEmains unclEar.MEthodsIn this study, thE ADC, Rituximab-MonomEthyl Auristatin E (MMAE) with a ValinE–CitrullinE clEavablE linkEr, was dEsignEd to invEstigatE its thErapEutic Efficacy against non-Hodgkin lymphoma (NHL) as wEll as thE undErlying mEchanisms. MEthylthiazolyldiphEnyl-tEtrazolium bromidE (MTT) was usEd to dEtEct growth inhibition in B-cEll lymphoma cEll linEs, Ramos and Daudi cElls, which wErE trEatEd by Rituximab-MMAE alonE or combinEd with autophagy conditionEr. Apoptosis was dEtEctEd by flow cytomEtry and immunohistochEmistry, and apoptosis inhibitor was EmployEd to discovEr thE rElationship bEtwEEn autophagy and apoptosis during thE Rituximab-MMAE trEatmEnt. Autophagy was dEtErminEd by thrEE standard tEchniquEs which includEd confocal microscopE, transmission ElEctron microscopE, and wEstErn blots. Ramos xEnograft tumors in BALB/c nudE micE wErE EstablishEd to invEstigatE thE antitumor EffEct of combination usE of Rituximab-MMAE and autophagy conditionEr in B-NHL thErapy.REsultsOur rEsults showEd that Rituximab-MMAE ElicitEd caspasE-3-dEpEndEnt apoptosis in NHL cElls and ExhibitEd potEnt thErapEutic Efficacy in vivo. Autophagy, which was charactErizEd by uprEgulatEd light chain 3-II ExprEssion, and accumulation of autophagosomEs, was triggErEd during thE Rituximab-MMAE trEatmEnt. MEanwhilE, inactivation of Akt/mTOR pathway was shown to bE involvEd in thE autophagy triggErEd by Rituximab-MMAE, indicating a probablE mEchanism of thE ADC-initiatEd autophagy. Importantly, inhibition of autophagy by chloroquinE supprEssEd thE Rituximab-MMAE-inducEd apoptosis, whilE activating autophagy by rapamycin significantly EnhancEd thE thErapEutic EffEct of Rituximab-MMAE both in vitro and in vivo.ConclusionOur data ElucidatEd thE critical rElationship bEtwEEn autophagy and apoptosis in Rituximab-MMAE-basEd thErapy and highlightEd thE potEntial approach for NHL thErapy by combinEd administration of thE ADC and autophagy activator