Herman A. Perroud

Therapeutic efficacy of metronomic chemotherapy with cyclophosphamide and doxorubicin on murine mammary adenocarcinomas

BACKGROUND Metronomic chemotherapy (MCT) refers to the chronic and equally spaced administration of low doses of different chemotherapy drugs, without extended rest periods. Herein, we investigated the therapeutic efficacy of metronomic cyclophosphamide (Cy) combined with doxorubicin (Dox) in two mouse mammary adenocarcinoma models. MATERIALS AND METHODS Mice were s.c. challenged with M-234p or M-406 mammary tumors, and when the tumors reached ∼150 mm(3), they were treated with: (I) no treatment (controls); (II) Cy in the drinking water (30 mg/kg body weight/day); (III) Dox (0.5 mg/kg body weight i.p. three times/week); (IV) treated as (II) + (III). Mice challenged i.v. with M-234p or M-406 tumor cells received, on day 3, the same treatments. RESULTS We found that MCT with Cy plus Dox inhibited tumor growth, decreased lung metastases, and increased the median survival time, while having low toxic effect. Combined MCT was more effective than each monotherapy causing decrease in VEGF serum concentration and tumor proliferation rate plus increase in tumor apoptosis. CONCLUSION(S) The therapeutic benefits of combined MCT with Cy and Dox on mammary adenocarcinomas together with its low toxic effect profile suggest the possibility of future translation into the clinic.

Association between baseline VEGF/sVEGFR-2 and VEGF/TSP-1 ratios and response to metronomic chemotherapy using cyclophosphamide and celecoxib in patients with advanced breast cancer

BACKGROUND Metronomic chemotherapy (MCT) with cyclophosphamide (Cy) and celecoxib (Cel) has therapeutic efficacy and low toxicity profile in advanced breast cancer patients (ABCP), but no reliable biomarkers of response have been found yet that allow patient selection for treatment. AIM To investigate the potential role as biomarkers of pro- and antiangiogenic parameters and evaluate their response in ABCP receiving metronomic Cy 50 mg p.o./day + Cel 400 mg p.o./day. MATERIALS AND METHODS Serum levels of vascular endothelial growth factor-C (VEGF-C), soluble VEGF receptors 2 and 3 (sVEGFR-2, sVEGFR-3), were measured at different time points in 13/15 patients included in a phase II trial of MCT with Cy+Cel. RESULTS Serum levels of sVEGFR-2 and sVEGFR-3 increased significantly during treatment (P = 0.0392; P = 0.0066, respectively). VEGF-C showed no significant modifications. Previous determinations of VEGF and TSP-1 in the same patients were utilized. VEGF/sVEGFR-2, VEGF/TSP-1, and VEGF-C/sVEGFR-3 ratios decreased significantly along the treatment (P = 0.0092; P = 0.0072; P = 0.0141, respectively). Nonsignificant variations were observed for VEGF-C/sVEGFR-2 ratio. Baseline values of VEGF/sVEGFR-2 and VEGF/TSP-1 ratios were associated with time to progression (TTP) (P = 0.0407; P = 0.0394, respectively) meanwhile baseline VEGF was marginally significant (P = 0.0716). Patients with values lower than the 50th percentile for both ratios showed longer TTP. CONCLUSIONS We have identified the baseline VEGF/sVEGFR-2 and VEGF/TSP-1 ratios as potential biomarkers of response in ABCP treated metronomically with Cy+Cel. This finding warrants its confirmation in a higher number of patients.

Comparative Effectiveness of Two Metronomic Chemotherapy Schedules—Our Experience in the Preclinical Field

Metronomic chemotherapy refers to the chronic, equally spaced, delivery of low doses of chemotherapeutic drugs, without extended interruptions. Previously, we developed two combined metronomic schemes for the treatment of murine mammary tumors. The aim of this study was to compare their effects on tumor and metastasis growth, survival, and toxicity. Metronomic chemotherapy with Cyclophosphamide + Celecoxib (Cy + Cel) showed higher antimetastatic power than Cyclophosphamide + Doxorubicin (Cy + Dox), while being similar in other aspects. That difference, plus the advantage that represents its oral administration, suggests that the Cy + Cel combination is more suitable than Cy + Dox for metronomic chemotherapy of mammary tumors and could be proposed to the translation to the clinic.

Clinical response in patients with ovarian cancer treated with metronomic chemotherapy

Ovarian cancer (OC) is the leading cause of death from gynaecological cancer. It is extremely hard to diagnose in the early stages and around 70% of patients present with advanced disease. Metronomic chemotherapy (MCT) is described as the chronic administration of, generally low, equally spaced, doses of chemotherapeutic drugs with therapeutic efficacy and low toxicity. This is an effective and low-cost way to treat several types of tumours, including ovarian cancer. Here, we present six cases of advanced ovarian cancer treated with MCT with low doses of cyclophosphamide, which showed clinical response and stable disease.

Putative Biomarkers of Response to Treatment in Breast Cancer Patients: A Pilot Assay

ABSTRACT Identifying tumor biomarkers associated with clinical behavior in breast cancer patients may allow higher accuracy in the selection of treatment. Different types of cells were determined in the primary tumors of stage I, II, and III of breast cancer patients, who were assigned to one of the two groups: (1) disease-free or (2) relapsed/progressed, at 5 years after primary treatment. We studied 32 tumor samples. CD4+ lymphocytes and CD44+CD24−/low cells (cancer stem cells) showed a significant association with clinical outcome at 5 years of primary treatment, while CD8+, Foxp3+, CD34+, and myeloid-derived suppressor cells did not show any association. Coincident with the results of individual analysis, we identified CD4+ cells and CD44+CD24−/low cells as good predictors of long-term clinical outcome in a logistic regression.

Quality of life in patients with metastatic breast cancer treated with metronomic chemotherapy

AIM The objective of the study was to detect changes in quality of life (QoL) in metastatic breast cancer patients treated with metronomic chemotherapy with daily low doses of cyclophosphamide and celecoxib. MATERIAL & METHODS Patients included in a Phase II trial, treated with metronomic cyclophosphamide and celecoxib were included in the QoL study. Assessment of QoL was carried out every 2 months by the Functional Assessment of Cancer Therapy Breast (FACT-B) questionnaire, Brief Pain Inventory and Eastern Cooperative Oncologic Group scale. Data were analyzed at three time points: baseline (BL); middle of treatment (MT); and end of treatment (ET). RESULTS A total of 20 patients were included. All patients were heavily pretreated. Treatment showed a good and safe therapeutic profile. With FACT-B questionnaire, no significant differences were observed during the response period (BL-MT). However, a significant increase was observed in the Emotional well-being and Additional concerns axes, when the last time point was included in the analysis (BL-MT-ET). A significant decrease in the proportion of patients with pain was found when comparing BL with ET (p = 0.046). The assessment with Eastern Cooperative Oncologic Group scale showed that 26.7% (4/15) of the patients improved their functional status and 40% (6/15) showed no changes, while 33.3% (5/10) worsened it. CONCLUSION Patients treated metronomically for several months did not worsen their QoL. A high proportion of patients showed improvement or no changes and there were less patients with pain at the end of the treatment.

Abstract P3-07-61: Predictors of response and follow up biomarkers for metronomic chemotherapy with cyclophosphamide and celecoxib in advanced breast cancer patients

Low-dose metronomic chemotherapy (MC) with Cyclophosphamide (Cy) and Celecoxib (Cel) has demonstrated to be effective and well-tolerated in advanced breast cancer patients (ABCP) but predictive markers of response or follow-up are lacking. Given the antiangiogenic properties of MC we analyzed several angiogenesis-related biomarkers and evaluated their potential role as predictors of response or treatment follow-up of ABCP treated with MC. Treatment plan: Patients received Cy 50 mg p.o./day + Cel 200 mg p.o./ bid. Cellular parameters: Circulating endothelial cells (CEC) and Circulating progenitor endothelial Cells (CEP) were determined by Flow Cytometry. Serologic parameters: Serum levels of vascular endothelial growth factor (VEGF), VEGF-C, soluble VEGF Receptors 2 and 3 (sVEGFR-2, sVEGFR-3) and Thrombospondin-1 (TSP-1) were determined by ELISA. Blood samples were collected before and during treatment. Twenty patients were enrolled. Response Rate was 5% and Clinical Benefit (CB) 55%. Most of the patients showed prolonged stable disease (SD≥24 weeks). Biomarkers were determined in all patients. Levels of CEC and CEP showed no clear trend variations during treatment. However, levels of CEC significantly increased at the time of disease progression in those patients who showed CB (P=0.014). Also baseline values of CEC and CEP showed marginally significant associations withTime To Progression. Serum VEGF concentration decreased during treatment (P=0.050) while sVEGFR-2 increased (P=0.005). VEGF-C, sVEGFR-3 and TSP-1 showed non-significant variations. VEGF/sVEGFR-2 ratio decreased during treatment (P=0.041), whereas VEGF/TSP-1, and VEGF-C/sVEGFR-2 ratios showed non-significant variations. Baseline values of VEGF, and VEGF/sVEGFR-2 showed negative and significant associations with TTP (P=0.0354 and P=0.0300, respectively) while sVEGFR-2 did not. When considering the two variables together, the goodness of prediction was not improved. To confirm the value of baseline VEGF and VEGF/sVEGFR-2 as predictors of response, we used the 50th percentile as a cutoff value to analyze the % of progression free survival. Patients with values lower than the 50th percentile for both biomarkers showed longer TTP (P=0.0001 and P=0.0008, respectively). The treatment had anti-angiogenic effect (VEGF decrease and sVEGFR-2 increase). The absence of variation in VEGF-C and sVEGFR-3 would indicate the lack of effect on lymphangiogenesis. Increased levels of CEC could be useful for detecting progression. If confirmed with a higher number of patients, baseline VEGF and VEGF/sVEGFR-2 values could be useful as early predictors of response. Citation Format: Perroud HA, Alasino CM, Rico MJ, Menacho-Marquez MA, Mainetti LE, Queralt F, Pezzotto SM, Rozados VR, Scharovsky G. Predictors of response and follow up biomarkers for metronomic chemotherapy with cyclophosphamide and celecoxib in advanced breast cancer patients. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P3-07-61.

Abstract 795: Metronomic chemotherapy with cyclophosphamide and metformin inhibits tumor and metastasis growth M-406 murine mammary adenocarcinoma

Proceedings: AACR Annual Meeting 2014; April 5-9, 2014; San Diego, CA Metronomic chemotherapy (MCT) refers to the chronic, equally spaced, delivery of low doses of different chemotherapeutic drugs, without extended interruptions. We have already demonstrated that MCT, as an intervention strategy, of metronomic Cy combined with celecoxib or doxorubicin inhibits mammary tumors growth. Our present aim was to analyze the antitumor and antimetastatic effects of MCT of Cy combined with Met. Inbred CBi mice were s.c. or i.v. challenged with M-406 tumor (Day 0), and treated when the tumors reached a volume of 100 a 150 mm3 (s.c.) or from Day 3 (i.v.) on, with: I) no further treatment (controls); II) Cy in the drinking water (20-30 mg/kg body weight/day); III) Met 3 times/week by gavage (100 mg/kg); IV) Treated as II + III. Tumors were measured and animals weighed twice a week and blood samples for glycemia determination were taken. On day 36 tumor volume in group IV (mm3, mean ± SEM: 929.7 ± 213.8) was lower than in groups I (2880.0 ± 1530.0), II (2896.4 ± 976.9) and III (3772.0 ± 68.0). On day 46, with only two groups remaining, tumor volume in group IV was lower than that in group II (p=0.032). All the groups differed in tumor volume doubling time (days, mean ± SEM; I: 3.8 ± 0.3, II: 5.3 ± 0.7, III: 3.6 ± 0.4, IV: 5.4 ± 0.2) (p<0.05). All the groups differed in survival (days, median, I: 30.5; II: 44; III: 29; IV: 50) (p=0.0063), showing group IV the highest one. There were neither weight losses nor changes in the glycemia with respect to basal values both in s.c. and i.v. experiments. In the i.v. experiment, animals were sacrificed on day 13, when the first mouse showed signs of metastatic illness. The number and diameter of lung metastasis were determined and total metastatic volume (TMV) was calculated. The experimental groups differed in TMV (p<0.0001), being those of groups II (mm3, mean ± SEM: 162.1 ± 41.4) and IV (218.6 ± 31.3) significantly lower (p<0.001 and p<0.01, respectively) than those in groups I (491.3 ± 80.6) and III (432.3 ± 54.5). These results clearly show that metronomic chemotherapy with a combination of cyclophosphamide and metformin significantly inhibits tumor growth and increase survival, also decreasing the development of metastasis of a murine mammary adenocarcinoma, while being devoid of toxicity. Citation Format: Antonela S. Asad, Jesus Basualdo, Lucia Micheletti, Maria C. Capello Gardenal, Herman A. Perroud, Maria J. Rico, Viviana R. Rozados, O. Graciela Scharovsky. Metronomic chemotherapy with cyclophosphamide and metformin inhibits tumor and metastasis growth M-406 murine mammary adenocarcinoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 795. doi:10.1158/1538-7445.AM2014-795

Abstract 4665: VEGF/SVEGFR-2 and VEGF/TSP-1 ratios as probable predictors of response to metronomic chemotherapy with Cyclophosphamide and Celecoxib in advanced breast cancer patients.

Proceedings: AACR 104th Annual Meeting 2013; Apr 6-10, 2013; Washington, DC The efficacy and low toxicity of antiangiogenic metronomic chemotherapy (MCT) with cyclophosphamide (Cy) and celecoxib (Cel) in advanced breast cancer patients (ABCP) was demonstrated in our studies but no reliable biomarkers or predictors of response have been found yet. The aim of this study was to analyze several pro- and anti-angiogenic parameters and evaluate their potential role as predictor of response duration in ABCP treated with MCT with daily Cy and Cel. Treatment plan: Patients received Cy 50 mg p.o./day + Cel 200 mg p.o. bid. This clinical trial was authorized by the School of Medicine Bioethics Committee and by A.N.M.A.T. (Argentine Regulatory Authority). .Angiogenesis parameters: Serum levels of vascular endothelial growth factor (VEGF), VEGF-C, soluble VEGF Receptors 2 and 3 (sVEGFR-2, sVEGFR-3) and Thrombospondin 1 (TSP-1) were determined by ELISA. Blood samples were collected before and during treatment and parametric tests were used to analyze data. Fifteen patients were enrolled, A partial response (PR) was observed in 1 patient (6.7%), which lasted 6 weeks. Prolonged Stable disease (pSD ≥24 weeks) was observed in 6/15 patients (40%). Median TTP among patients with pSD was 37.5 weeks (range: 26.43-81.57). The OCB obtained was 46.7% (PR=1/15 + SD=6/15). The median TTP (PR+pSD) was 33 weeks (range: 10.43-81.57). Treatment toxicity was very low and no grade 3 or 4 adverse events were registered. Biomarkers were determinated in 13 patients. Serum VEGF concentration decreased as a function of time (P=0.004); sVEGFR-2 increased during response (P= 0.0268) while non-significant variations were detected in VEGF-C, sVEGFR-3 and TSP-1. The baseline values of VEGF, and VEGF/sVEGFR-2 and VEGF/TSP-1 ratios were significantly correlated with response duration (P=0.029, P=0.015, P=0.014, respectively). To assess which of the three variables measured at the beginning of the treatment could be used to predict response duration, linear regression analyses were done. VEGF/sVEGFR-2 and VEGF/TSP-1 ratios were good predictors (P=0.028, P=0.029, respectively), meanwhile VEGF was not (P=0.059). When considering VEGF, VEGF/VEGFR-2 and VEGF/TSP-1 in a multiple regression analysis, the goodness of prediction was not improved with respect to that obtained with each putative predictor. We conclude that the antiangiogenic nature of MCT with Cy plus Cel is confirmed through the decrease of VEGF and the increase of sVEGFR-2; the absence of variation in VEGF-C and sVEGFR-3 would indicate the lack of effect on lymphangiogenesis; if confirmed in a higher number of patients, VEGF/sVEGFR-2 and VEGF/TSP-1 ratios could be useful as early predictors of treatment response. Citation Format: Herman A. Perroud, Carlos M. Alasino, Maria J. Rico, Francisco Queralt, Stella M. Pezzotto, Viviana R. Rozados, O.Graciela Scharovsky. VEGF/SVEGFR-2 and VEGF/TSP-1 ratios as probable predictors of response to metronomic chemotherapy with Cyclophosphamide and Celecoxib in advanced breast cancer patients. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4665. doi:10.1158/1538-7445.AM2013-4665

Abstract 2060: Metronomic chemotherapy (MCT): comparison of the efficacy of two combinations of drugs for the treatment of murine mammary adenocarcinomas.

Metronomic chemotherapy (MCT) refers to the chronic, equally spaced, delivery of low doses of different chemotherapeutic drugs, without extended interruptions. We have developed two combined MCT schemes for the treatment of mammary tumors: A) cyclophosphamide (Cy) 30 mg/kg/day p.o. + celecoxib (Cel) 30 mg/kg p.o. 5 times/week, and B) Cy 20 mg/kg/day p.o. + doxorubicin (Dox) 0.5 mg/kg, 3 times/week i.p. They were tested in two experimental models of mammary adenocarcinomas: I) M-406 and II) M-234p syngeneic with inbred CBi and BALB/c mice, respectively. Our present aim was to compare efficacy and toxicity of MCT combined A and B treatments in both tumor-models. There were analyzed antitumor and antimetastatic effects, survival and toxicity. The % of reduction of tumor volume of animals in the treated group with respect to control group without treatment [median (range): AI: 77.9 (50.9-89.9), AII: 84.4 (30-99.4), BI: 95.6 (57-99.6), BII: 75.5 (62.5-96.8)] didn9t differ between treatments or between tumor-models (non parametric ANOVA). The % of reduction of lung metastatic burden [AI: 99.8 (97.1-99.2) AII: 99.7 (98.7-100), BI: 90.6 (35.9-96.6), BII: 90 (48.4-99.3)] was significantly different among all the groups (P Citation Format: Maria J. Rico, Leandro E. Mainetti, Herman A. Perroud, Viviana R. Rozados, O. Graciela Scharovsky. Metronomic chemotherapy (MCT): comparison of the efficacy of two combinations of drugs for the treatment of murine mammary adenocarcinomas. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2060. doi:10.1158/1538-7445.AM2013-2060