In contrast, the full total daily dose for the XL184 PD-BEDR is slightly higher than the best dose been shown to be tolerable in posted xenograft research (26) and approximately twice the mouse exact carbon copy of the individual MTD (22); hence, despite our data demonstrating no significant bodyweight loss through the XL184 PD-BEDR, thorough toxicology analysis will be necessary to ensure tolerability of the regimen fully. Despite the multi-targeting nature of the MET TKIs examined here, our PD-BEDRs based exclusively on MET kinase inhibition conferred tumor regression in the SNU-5 xenograft model. yielded substantial and sustained tumor regression; the equivalent effectiveness of customized dosage regimens that achieve the same level of continuous molecular target control represents preclinical proof-of-concept and illustrates the importance of proper scheduling of targeted agent BEDs. PD-guided Biologically Effective Dosage Regimens (PD-BEDRs) potentially offer a superior alternative to pharmacokinetic guidance (e.g., drug concentrations in surrogate tissues) for developing and making head-to-head comparisons of targeted agents. suppress pY1234/1235MET/MET by 90%a level that was selected based on a study of PF02341066 (crizotinib) that equated > 90% pMET suppression with tumor growth inhibition, but not regression, in a MET-amplified preclinical model (17). Furthermore, by examining pharmacodynamic measurements from a small number of clinically feasible time points, we have modeled how this PD biomarker endpoint could be used as a primary endpoint of a clinical Phase 0 trial comparing depth and duration of the molecular response to safe single doses of several agents, with patient cohorts representing different time points after drug administration. Our results demonstrate the effectiveness of pharmacodynamically-guided, biologically effective dosage regimens in achieving sustained molecular target control and, thereby, antitumor efficacy, supporting proof-of-concept evaluation of this approach in the clinic. MATERIALS AND METHODS Therapeutic agents MET TKIs EMD1214063 (NSC 758244, tepotinib), ARQ197 (NSC 758242, tivantinib), XL184 (NSC 761068, cabozantinib), and XL880 (NSC 755775, GSK1363089, active ingredient in foretinib) were obtained from the Developmental Therapeutics Program, National Cancer Institute (NCI). Chemical structures are shown in Supplementary Fig. S1. Agent purity was confirmed by proton-carbon NMR, HPLC, and mass spectrometry. Animal models and drug administration The Frederick National Laboratory for Cancer Research (FNLCR) is accredited by the Association for Assessment and Accreditation of Laboratory Animal Care International and follows the Public Health Service Policy for the Care and Use of Laboratory Animals. All studies were conducted according to an approved animal care and use committee protocol in accordance with procedures outlined in the Guide for Care and Use of Laboratory Animals 8th Edition (National Research Council; 2011; The National Academies Press; Washington, D.C.). Female athymic nu/nu (NCr) mice (NCI Animal Production Program, Frederick, MD) were implanted subcutaneously with SNU-5 or GTL-16 human gastric tumor cells (16). Mice were housed in sterile, filter-capped, polycarbonate cages (Allentown Caging, Allentown, NJ) maintained in a barrier facility on a 12-hour light/dark cycle and were provided sterilized food and water ad libitum. Mice were randomized into groups before initiation of treatment using a commercial software program (Study Director, Studylog Systems, Inc., South San Francisco, CA). Single-dose PK/PD study After tumors reached 200 mm3, mice were orally administered with MET inhibitor at doses equivalent to the human MTDs, as known at the time of study design (20,28,29), and 1/3, 1/6, and 1/10 MTD; as the MTD was unknown for EMD1214063 at the time of this study, we used doses that were shown in the literature to be active (27,30). Doses were: EMD1214063 at 1, 3, 10, and 30 mg/kg; ARQ197 at 6, 24, 80, and 240 mg/kg; XL184 at 3.3, 5.5, 11, and 33 mg/kg; and XL880 at 8.3, 14, 28, and 83 mg/kg. Providers were prepared as follows: XL880, 0.75% hydroxypropyl methylcellulose (HPMC)/0.15% sodium lauryl sulfate (SLS) in distilled water; EMD1214063, 10% dimethyl sulfoxide (DMSO) in saline; XL184, distilled water; ARQ197, polyethylene glycol 400: 20% vitamin E-d- tocopheryl polyethylene glycol 1000 succinate (TPGS) answer (60:40). Plasma and tumor samples (= 3/time point) collected 0.5, 1, 2, 4, 6, 12, and 24 hours after drug administration were adobe flash frozen for pharmacokinetic and pharmacodynamic analyses. Multiple-dose PD-BEDR effectiveness assessment Mice were given oral doses of XL880 (17 mg/kg once daily), XL184 (44 mg/kg twice daily), or EMD1214063 (12.5 mg/kg twice daily) for 21 days once tumors reached 150 mm3. These dose regimens were selected based on PD data from.The time course and magnitude of pY1234/1235MET/MET reduction varied considerably among MET inhibitors, so we defined the BED for each by identifying the minimal single dose required to achieve > 90% reduction in tumor pY1234/1235MET/METas well as the duration of that effect. malignancy xenograft model (SNU-5). Reductions in tumor pY1234/1235MET/total MET of 95C99% were attainable with tolerable doses of EMD1214063/MSC2156119J (tepotinib), XL184 (cabozantinib), and XL880/GSK1363089 (foretinib), but not ARQ197 (tivantinib), which did not alter the PD biomarker. Duration of kinase suppression and rate of kinase recovery were specific to each agent, emphasizing the importance of developing customized dose regimens to accomplish continuous suppression of the PD biomarker at the required level (here, 90% MET kinase suppression). The customized dose routine of each inhibitor yielded considerable and sustained tumor regression; the equivalent performance of customized dose regimens that accomplish the same level of continuous molecular target control signifies preclinical proof-of-concept and illustrates the importance of proper scheduling of targeted agent Mattresses. PD-guided Biologically Effective Dosage Regimens (PD-BEDRs) potentially offer a superior alternative to pharmacokinetic guidance (e.g., drug concentrations in surrogate cells) for developing and making head-to-head comparisons of targeted providers. suppress pY1234/1235MET/MET by 90%a level that was selected based on a study of PF02341066 (crizotinib) that equated > 90% pMET suppression with tumor growth inhibition, but not regression, inside a MET-amplified preclinical model (17). Furthermore, by analyzing pharmacodynamic measurements from a small number of clinically feasible time points, we have modeled how this PD biomarker endpoint could be used like a main endpoint of a clinical Phase 0 trial comparing depth and period of the molecular response to safe single doses of several providers, with patient cohorts representing different time points after drug administration. Our results demonstrate the effectiveness of pharmacodynamically-guided, biologically effective dose regimens in achieving sustained molecular target control and, therefore, antitumor efficacy, assisting proof-of-concept evaluation of this approach in the medical center. MATERIALS AND METHODS Therapeutic providers MET TKIs EMD1214063 (NSC 758244, tepotinib), ARQ197 (NSC 758242, tivantinib), XL184 (NSC 761068, cabozantinib), and XL880 (NSC 755775, GSK1363089, active ingredient in foretinib) were from the Developmental Therapeutics System, National Malignancy Institute (NCI). Chemical constructions are shown in Supplementary Fig. S1. Agent purity was confirmed by proton-carbon NMR, HPLC, and mass spectrometry. Animal models and drug administration The Frederick National Laboratory for Cancer Study (FNLCR) is accredited from the Association for Assessment and Accreditation of Laboratory Animal Care International HDAC6 and follows the Public Health Service Policy for the Care and Use of Laboratory Animals. All studies were conducted relating to an authorized animal care and attention and use committee protocol in accordance with procedures layed out in the Guideline for Care and Use of Laboratory Animals 8th Release (National Study Council; 2011; The National Academies Press; Washington, D.C.). Female athymic nu/nu (NCr) mice (NCI Animal Production System, Frederick, MD) were implanted subcutaneously with SNU-5 or GTL-16 human being gastric tumor cells (16). Mice were housed in sterile, filter-capped, polycarbonate cages (Allentown Caging, Allentown, NJ) managed in a barrier facility on a 12-hour light/dark cycle and were provided sterilized food and water ad libitum. Mice were randomized into groups before initiation of treatment using a commercial software program (Study Director, Studylog Systems, Inc., South San Francisco, CA). Single-dose PK/PD study After tumors reached 200 mm3, mice were orally administered with MET inhibitor at doses equivalent to the human MTDs, as known at the time of study design (20,28,29), and 1/3, 1/6, and 1/10 MTD; as the MTD was unknown for EMD1214063 at the time of this study, we used doses that were shown in the literature to be active (27,30). Doses were: EMD1214063 at 1, 3, 10, and 30 mg/kg; ARQ197 at 6, 24, 80, and 240 mg/kg; XL184 at 3.3, 5.5, 11, and 33 mg/kg; and XL880 at 8.3, 14, 28, and 83 mg/kg. Brokers were prepared as follows: XL880, 0.75% hydroxypropyl methylcellulose (HPMC)/0.15% sodium lauryl sulfate (SLS) in distilled water; EMD1214063, 10% dimethyl sulfoxide (DMSO) in saline; XL184, distilled water; ARQ197, polyethylene glycol 400: 20% vitamin.Standard criteria for acceptable accuracy and reproducibility were applied. XL184 (cabozantinib), and XL880/GSK1363089 (foretinib), but not ARQ197 (tivantinib), which did not alter the PD biomarker. Duration of kinase suppression and rate of kinase recovery were specific to each agent, emphasizing the importance of developing customized dosage regimens to achieve continuous suppression of the PD biomarker at the required level (here, 90% MET kinase suppression). The customized dosage regimen of each inhibitor yielded substantial and sustained tumor regression; the equivalent effectiveness of customized dosage regimens that achieve the same level of continuous molecular target control represents preclinical proof-of-concept and illustrates the importance of proper scheduling of targeted agent BEDs. PD-guided Biologically Effective Dosage Regimens (PD-BEDRs) potentially offer a superior alternative to pharmacokinetic guidance (e.g., drug concentrations in surrogate tissues) for developing and making head-to-head comparisons of targeted brokers. suppress pY1234/1235MET/MET by 90%a level that was selected based on a study of PF02341066 (crizotinib) that equated > 90% pMET suppression with tumor growth inhibition, but not regression, in a MET-amplified preclinical model (17). Furthermore, by examining pharmacodynamic measurements from a small number of clinically feasible time points, we have modeled how this PD biomarker endpoint could be used as a primary endpoint of a clinical Phase 0 trial comparing depth and duration of the molecular response to safe single doses of several brokers, with patient cohorts representing different time points after drug administration. Our results demonstrate the effectiveness of pharmacodynamically-guided, biologically effective dosage regimens in achieving sustained molecular target control and, thereby, antitumor efficacy, supporting proof-of-concept BRL-50481 evaluation of this approach in the clinic. MATERIALS AND METHODS Therapeutic brokers MET TKIs EMD1214063 (NSC 758244, tepotinib), ARQ197 (NSC 758242, tivantinib), XL184 (NSC 761068, cabozantinib), and XL880 (NSC 755775, GSK1363089, active ingredient in foretinib) were obtained from the Developmental Therapeutics Program, National Malignancy Institute (NCI). Chemical structures are shown in Supplementary Fig. S1. Agent purity was confirmed by proton-carbon NMR, HPLC, and mass spectrometry. Animal models and drug administration The Frederick National Laboratory for Cancer Research (FNLCR) is accredited by the Association for Assessment and Accreditation of Laboratory Animal Care International and follows the Public Health Service Policy for the Care and Use of Laboratory Animals. All research were conducted BRL-50481 relating for an authorized animal care and attention and make use of committee protocol relative to procedures defined in the Guidebook for Treatment and Usage of Lab Animals 8th Release (National Study Council; 2011; The Country wide Academies Press; Washington, D.C.). Feminine athymic nu/nu (NCr) mice (NCI Pet Production System, Frederick, MD) had been implanted subcutaneously with SNU-5 or GTL-16 human being gastric tumor cells (16). Mice had been housed in sterile, filter-capped, polycarbonate cages (Allentown Caging, Allentown, NJ) taken care of in a hurdle facility on the 12-hour light/dark routine and were offered sterilized water and food advertisement libitum. Mice had been randomized into organizations before initiation of treatment utilizing a commercial computer software (Study Movie director, Studylog Systems, Inc., South SAN FRANCISCO BAY AREA, CA). Single-dose PK/PD research After tumors reached 200 mm3, mice had been orally given with MET inhibitor at dosages equal to the human being MTDs, as known during study style (20,28,29), and 1/3, 1/6, and 1/10 MTD; as the MTD was unfamiliar for EMD1214063 during this research, we used dosages that were demonstrated in the books to be energetic (27,30). Dosages had been: EMD1214063 at 1, 3, 10, and 30 mg/kg; ARQ197 at 6, 24, 80, and 240 mg/kg; XL184 at 3.3, 5.5, 11, and 33 mg/kg; and XL880 at 8.3, 14, 28, and 83 mg/kg. Real estate agents were prepared the following: XL880, 0.75% hydroxypropyl methylcellulose (HPMC)/0.15% sodium lauryl sulfate (SLS) in distilled water; EMD1214063, 10% dimethyl sulfoxide (DMSO) in saline; XL184, distilled drinking water; ARQ197, polyethylene glycol 400: 20% supplement E-d- tocopheryl polyethylene glycol 1000 succinate (TPGS) remedy (60:40). Plasma and tumor examples (= 3/period point) gathered 0.5, 1, 2, 4, 6, 12, and a day after medication administration were adobe flash frozen for pharmacokinetic and pharmacodynamic analyses. Multiple-dose PD-BEDR effectiveness assessment Mice received oral dosages of XL880 (17 mg/kg once daily), XL184 (44 mg/kg double daily), or EMD1214063 (12.5 mg/kg twice daily) for 21 times once tumors reached 150 mm3. These dose regimens were chosen predicated on PD data through the single-dose study; we used relevant dosage increment ratios of just one 1 clinically.33 or 1.2 to escalate through the dosage level that had yielded slightly significantly less than sufficient pY1234/1235MET/MET suppression in the single-dose research. Tumor quantities had been documented up to review day time 62 daily, and tumor development inhibition was dependant on percent treatment/control (%T/C) (31). PD-based dose regimen marketing for EMD1214063 For single-dose PD biomarker assessments,.We foresee the use of this process to additional therapeutic classes of molecularly targeted real estate agents. Supplementary Material 1Click here to see.(415K, docx) Acknowledgments This project continues to be funded entirely or partly with federal funds through the National Cancer Institute, National Institutes of Health, under Contract No. model (SNU-5). Reductions in tumor pY1234/1235MET/total MET of 95C99% had been attainable with tolerable dosages of EMD1214063/MSC2156119J (tepotinib), XL184 (cabozantinib), and XL880/GSK1363089 (foretinib), however, not ARQ197 (tivantinib), which didn’t alter the PD biomarker. Duration of kinase suppression and price of kinase recovery had been particular to each agent, emphasizing the need for developing customized dose regimens to accomplish constant suppression from the PD biomarker at the mandatory level (right here, 90% MET kinase suppression). The personalized dose regimen of every inhibitor yielded considerable and suffered tumor regression; the same effectiveness of personalized dose regimens that obtain the same degree of constant molecular focus on control represents preclinical proof-of-concept and illustrates the need for proper arranging of targeted agent Bedrooms. PD-guided Biologically Effective Dosage Regimens (PD-BEDRs) possibly offer a excellent option to pharmacokinetic assistance (e.g., medication concentrations in surrogate tissue) for developing and producing head-to-head evaluations of targeted realtors. suppress pY1234/1235MET/MET by 90%a level that was chosen based on a report of PF02341066 (crizotinib) that equated > 90% pMET suppression with tumor development inhibition, however, not regression, within a MET-amplified preclinical model (17). Furthermore, by evaluating pharmacodynamic measurements from a small amount of clinically feasible period points, we’ve modeled how this PD biomarker endpoint could possibly be used being a principal endpoint of the clinical Stage 0 trial evaluating depth and length of time from the molecular response to secure single dosages of several realtors, with individual cohorts representing different period points after medication administration. Our outcomes demonstrate the potency of pharmacodynamically-guided, biologically effective medication dosage regimens in attaining sustained molecular focus on control and, thus, antitumor efficacy, helping proof-of-concept evaluation of the strategy in the medical clinic. MATERIALS AND Strategies Therapeutic realtors MET TKIs EMD1214063 (NSC 758244, tepotinib), ARQ197 (NSC 758242, tivantinib), XL184 (NSC 761068, cabozantinib), and XL880 (NSC 755775, GSK1363089, active component in foretinib) had been extracted from the Developmental Therapeutics Plan, National Cancer tumor Institute (NCI). Chemical substance buildings are shown in Supplementary Fig. S1. Agent purity was verified by proton-carbon NMR, HPLC, and mass spectrometry. Pet models and medication administration The Frederick Country wide Lab for Cancer Analysis (FNLCR) is certified with the BRL-50481 Association for Evaluation and Accreditation of Lab Animal Treatment International and comes after the Public Wellness Service Plan for the Treatment and Usage of Lab Animals. All research were conducted regarding to an accepted animal caution and make use of committee protocol relative to procedures specified in the Instruction for Treatment and Usage of Lab Animals 8th Model (National Analysis Council; 2011; The Country wide Academies Press; Washington, D.C.). Feminine athymic nu/nu (NCr) mice (NCI Pet Production Plan, Frederick, MD) had been implanted subcutaneously with SNU-5 or GTL-16 individual gastric tumor cells (16). Mice had been housed in sterile, filter-capped, polycarbonate cages (Allentown Caging, Allentown, NJ) preserved in a hurdle facility on the 12-hour light/dark routine and were supplied sterilized water and food advertisement libitum. Mice had been randomized into groupings before initiation of treatment utilizing a commercial computer software (Study Movie director, Studylog Systems, Inc., South SAN FRANCISCO BAY AREA, CA). Single-dose PK/PD research After tumors reached 200 mm3, mice had been orally implemented with MET inhibitor at dosages equal to the individual MTDs, as known during study style (20,28,29), and 1/3, 1/6, and 1/10 MTD; as the MTD was unidentified for EMD1214063 during this research, we used dosages that were proven in the books to be energetic (27,30). Dosages had been: EMD1214063 at 1, 3, 10, and 30 mg/kg; ARQ197 at 6, 24, 80, and 240 mg/kg; XL184 at 3.3, 5.5, 11, and 33 mg/kg; and XL880 at 8.3, 14, 28, and 83 mg/kg. Agencies were prepared the following: XL880, 0.75% hydroxypropyl methylcellulose (HPMC)/0.15% sodium lauryl sulfate (SLS) in distilled water; EMD1214063, 10% dimethyl sulfoxide (DMSO) in saline; XL184, distilled drinking water; ARQ197, polyethylene glycol 400: 20% supplement E-d- tocopheryl polyethylene glycol 1000 succinate (TPGS) option (60:40). Plasma and tumor examples (= 3/period point) gathered 0.5, 1, 2, 4, 6, 12, and a day after medication administration were display frozen for pharmacokinetic and pharmacodynamic analyses. Multiple-dose PD-BEDR efficiency assessment Mice received oral dosages of XL880 (17 mg/kg once daily), XL184 (44 mg/kg double daily), or EMD1214063 (12.5 mg/kg twice daily) for 21 times once tumors reached 150 mm3. These medication dosage regimens were chosen predicated on PD data in the single-dose research; we used medically relevant dosage increment ratios of just one 1.33 or 1.2 to escalate in the dosage level that had yielded slightly significantly less than sufficient pY1234/1235MET/MET suppression in the single-dose research. Tumor volumes had been documented daily up to review time 62, and tumor development inhibition was dependant on percent treatment/control.period data for every inhibitor using WinNonlin (Pharsight Corp., Hill View, CA), Edition 4.1 (ARQ197 and EMD-1214063) or Version 5.2 (XL184 and XL880), using non-compartmental choices. Statistical Analysis All descriptive statistics (including mean, SD, and CV), Learners unpaired test (2-tailed), and Fishers exact check were executed using Microsoft GraphPad and Excel Prism software program (v3.04, La Jolla, CA). PD biomarker. Duration of kinase suppression and price of kinase recovery had been particular to each agent, emphasizing the need for developing customized medication dosage regimens to attain constant suppression from the PD biomarker at the mandatory level (right here, 90% MET kinase suppression). The personalized medication dosage regimen of every inhibitor yielded significant and suffered tumor regression; the same effectiveness of personalized medication dosage regimens that BRL-50481 obtain the same degree of constant molecular focus on control represents preclinical proof-of-concept and illustrates the need for proper arranging of targeted agent Bedrooms. PD-guided Biologically Effective Dosage Regimens (PD-BEDRs) possibly offer a excellent option to pharmacokinetic assistance (e.g., medication concentrations in surrogate tissue) for developing and producing head-to-head evaluations of targeted agencies. suppress pY1234/1235MET/MET by 90%a level that was chosen based on a report of PF02341066 (crizotinib) that equated > 90% pMET suppression with tumor development inhibition, however, BRL-50481 not regression, within a MET-amplified preclinical model (17). Furthermore, by evaluating pharmacodynamic measurements from a small amount of clinically feasible period points, we’ve modeled how this PD biomarker endpoint could possibly be used being a principal endpoint of the clinical Stage 0 trial evaluating depth and length of time from the molecular response to secure single dosages of several agencies, with individual cohorts representing different period points after medication administration. Our outcomes demonstrate the potency of pharmacodynamically-guided, biologically effective medication dosage regimens in attaining sustained molecular focus on control and, thus, antitumor efficacy, helping proof-of-concept evaluation of the strategy in the medical clinic. MATERIALS AND Strategies Therapeutic agencies MET TKIs EMD1214063 (NSC 758244, tepotinib), ARQ197 (NSC 758242, tivantinib), XL184 (NSC 761068, cabozantinib), and XL880 (NSC 755775, GSK1363089, active component in foretinib) had been extracted from the Developmental Therapeutics Plan, National Cancers Institute (NCI). Chemical substance buildings are shown in Supplementary Fig. S1. Agent purity was verified by proton-carbon NMR, HPLC, and mass spectrometry. Pet models and medication administration The Frederick Country wide Lab for Cancer Analysis (FNLCR) is certified with the Association for Evaluation and Accreditation of Lab Animal Treatment International and comes after the Public Wellness Service Plan for the Treatment and Usage of Lab Animals. All research were conducted regarding to an accepted animal caution and make use of committee protocol relative to procedures discussed in the Information for Treatment and Usage of Lab Animals 8th Edition (National Research Council; 2011; The National Academies Press; Washington, D.C.). Female athymic nu/nu (NCr) mice (NCI Animal Production Program, Frederick, MD) were implanted subcutaneously with SNU-5 or GTL-16 human gastric tumor cells (16). Mice were housed in sterile, filter-capped, polycarbonate cages (Allentown Caging, Allentown, NJ) maintained in a barrier facility on a 12-hour light/dark cycle and were provided sterilized food and water ad libitum. Mice were randomized into groups before initiation of treatment using a commercial software program (Study Director, Studylog Systems, Inc., South San Francisco, CA). Single-dose PK/PD study After tumors reached 200 mm3, mice were orally administered with MET inhibitor at doses equivalent to the human MTDs, as known at the time of study design (20,28,29), and 1/3, 1/6, and 1/10 MTD; as the MTD was unknown for EMD1214063 at the time of this study, we used doses that were shown in the literature to be active (27,30). Doses were: EMD1214063 at 1, 3, 10, and 30 mg/kg; ARQ197 at 6, 24, 80, and 240 mg/kg; XL184 at 3.3, 5.5, 11, and 33 mg/kg; and XL880 at 8.3, 14, 28, and 83 mg/kg. Agents were prepared as follows: XL880, 0.75% hydroxypropyl methylcellulose (HPMC)/0.15% sodium lauryl sulfate (SLS) in distilled water; EMD1214063, 10% dimethyl sulfoxide (DMSO) in saline; XL184, distilled water; ARQ197, polyethylene glycol 400: 20% vitamin E-d- tocopheryl polyethylene glycol 1000 succinate (TPGS) solution (60:40). Plasma and tumor samples (= 3/time point) collected 0.5, 1, 2, 4, 6, 12, and 24 hours after drug administration were flash frozen for pharmacokinetic and pharmacodynamic analyses. Multiple-dose PD-BEDR efficacy assessment Mice were given oral doses of XL880 (17 mg/kg once daily), XL184 (44 mg/kg twice daily), or EMD1214063 (12.5 mg/kg twice daily) for 21 days once tumors reached 150 mm3. These dosage regimens were selected based on PD data from the single-dose study; we used clinically relevant dose increment ratios of 1 1.33 or 1.2 to escalate from the dose level that had yielded slightly less than adequate pY1234/1235MET/MET suppression in the single-dose study. Tumor volumes were recorded daily up to study day 62, and tumor growth inhibition was determined by percent treatment/control (%T/C) (31). PD-based dosage regimen optimization for EMD1214063 For single-dose PD biomarker assessments, mice bearing GTL-16 or SNU-5 tumors were given a.