Supplemental Table 3. Cardiac and Pituitary Analyses. FSH, follicle stimulating hormone; MUGA, multigated acquisition study; TSH, thyroid stimulating hormone.

e15153 Background: The prognosis for patients with castration-resistant prostate cancer (CRPC) remains suboptimal and targeted therapies should be explored. One potential target is the receptor for luteinizing hormone-releasing hormone (LHRH), which is highly expressed on prostate cancer cells. AEZS-108 is an LHRH-cytotoxic hybrid whose rational design covalently couples an LHRH agonist and the cytotoxic doxorubicin. AEZS-108 exploits the presence of LHRH receptors to target delivery of the cytotoxic. We report the phase I trial of AEZS-108 in men with taxane-resistant CRPC. We also report correlative studies of a novel circulating tumor cell (CTC) capture device that will provide enumeration of CTCs and results from AEZS-108 internalization studies that exploit the auto-fluorescence of doxorubicin in captured CTCs. Methods: This is a single-arm, dose-escalation phase I study in men with CRPC to confirm the dose established in a completed phase I trial in females. Eligibility criteria included adequate organ function and progression of disease despite prior therapy with an LHRH agonist and at least one taxane-based regimen. Patients were required to discontinue LHRH agonists to avoid receptor competition. Due to potential cardiotoxicity, patients with an ejection fraction < 50% or prior exposure to doxorubicin or mitoxantrone were excluded. Pituitary function was closely monitored. Patients received AEZS-108 every 21 days for up to 6 cycles until progression or unacceptable toxicity. The primary endpoint was safety. Results: Enrollment began in November 2010 and is ongoing. Currently, 13 patients have been enrolled. The first two planned dose levels had no dose-limiting toxicities observed. Two patients on the third dose level experienced a dose limiting toxicity. The second dose level has been reopened for expansion. There have been no cardiac or pituitary toxicities. At the time of submission, a decrease in PSA was noted in 6 of the 13 patients. Final results detailing safety, response and the suggested dose for the phase II portion will be reported. All correlative studies will also be reported. Conclusions: In general, AEZS-108 has been well tolerated. The phase II portion of the study will begin once the MTD is established.

Abstract Purpose: AEZS-108, formerly AN-152, is a cytotoxic hybrid molecule consisting of a luteinizing hormone-releasing hormone (LHRH) agonist moiety covalently coupled to doxorubicin, allowing it to deliver doxorubicin selectively to cells expressing LHRH receptors. LHRH receptors are expressed on the cell membrane of many tumors, including prostate cancer. This phase I study determined the maximum tolerated dose (MTD) of AEZS-108 in men with taxane- and castration-resistant prostate cancer (CRPC) while providing additional information on the safety profile and efficacy of this agent. Experimental Design: AEZS-108 was administered as an intravenous infusion every 21 days until progression or unacceptable toxicity in cohorts of 3 or 6 patients until the MTD was reached. Blood was collected for capture of circulating tumor cells (CTC) to visualize internalization of AEZS-108, an autofluorescent molecule. Results: The MTD of AEZS-108 in this cohort was 210 mg/m2, which was lower than that seen in a phase I study conducted in women with endometrial or ovarian cancers. The dose-limiting toxicity was persistent neutropenia. Three patients had a PSA response with an additional 10 patients maintaining PSA stable disease. Of the 10 patients evaluable by RECIST criteria, 9 achieved stable disease. AEZS-108 internalization in CTCs was routinely visualized using its autofluorescence. Conclusion: These findings show that AEZS-108 has an acceptable safety profile and a signal of efficacy, lowering PSA in heavily pretreated patients with prostate cancer, and that internalization of AEZS-108 in prostate cancer CTCs may be a viable pharmacodynamic marker. A phase II study in men with prostate cancer is ongoing. Clin Cancer Res; 20(24); 6277–83. ©2014 AACR.

e15596 Background: AEZS-108 (AN-152, zoptarelin doxorubicin) is an LHRH-cytotoxic hybrid drug consisting of a Luteinizing Hormone Releasing Hormone (LHRH) agonist coupled to the cytotoxic radical, doxorubicin. The binding permits AEZS-108 to accumulate on the surface of cells expressing LHRH receptors (LHRH-R) thus allowing drug uptake. This approach exploits LHRH-R to gain access to the targeted tumor cells. Once internalized, the cytotoxic properties of doxorubicin induce the anti-tumor response. Methods: This is a case report of a 66 year old male with a superficial bladder cancer history who was diagnosed in May 2009 with metastatic UC from the prostatic urethra. He presented with pelvic and retroperitoneal lymph nodes (LN). Patient had a partial response to cisplatin based chemotherapy and underwent radical cystoprostatectomy in Oct 2009. No residual UC in bladder or prostate was seen on pathology's examination. However he did have 11/24 removed LN positive for UC. The patient showed increased retroperitoneal LN by December 2009. He received 2 further lines of chemotherapy with no response and developed severe back pain, cervical LN, extensive liver metastases and a bulky retroperitoneal mass. The patient entered a Phase I clinical trial of AEZS-108, for “locally advanced unresectable or metastatic LHRH-R positive UC patients who failed platinum based chemotherapy“. He received AEZS-108 every 3 weeks for 9 cycles from May to October, 2011. Results: After 2 cycles of the investigational drug all pain and palpable lymph neck nodes disappeared; he stopped all narcotics and tolerated treatment very well. Mostly grade1-2 with few grade 3 toxicities were seen. CT scan pretreatment, and at 8, 22, 75 weeks post-treatment showed dramatic response at all disease sites. Twenty months after treatment start, the patient is symptom- free with no evidence of disease. Conclusions: This is the first case of a patient with LHRH-R positive Urothelial Carcinoma treated with AEZS-108 that is being reported. Considering the generally poor outcome of progressive UC and the short survival of these patients, the effect of this drug in this patient has been remarkable. The phase one study is ongoing.

Überexpression von ERK wurde in 34% der trripelnegativen Mammakarzinomen beschrieben und ist mit Anthtrazyklinresistenz assoziiert. (1). AEZS-131 ist ein oral wirksamer, hochselektiver Inhbitor von ERK ½. Der in in vivo Modellen des Kolonkarzinoms Antitumoraktivität gezeigt hat. Die vorliegende Studie untersucht den Wirkmechanismus und die Effektivität in in vitro Modellen des tripelnegativen Mammakarzinoms.

60 Background: The prognosis for patients with castration-resistant prostate cancer (CRPC) remains suboptimal and targeted therapies should be explored. One potential target is the receptor for luteinizing hormone-releasing hormone (LHRH-R), which is highly expressed on prostate cancer cells. AEZS-108 (AN-152) is an LHRH-cytotoxic hybrid whose rational design covalently couples an LHRH agonist and the cytotoxic doxorubicin. AEZS-108 exploits the presence of these receptors to target delivery of the cytotoxic. We report the phase I trial of AEZS-108 in men with taxane-resistant CRPC. We also report correlative studies of a novel circulating tumor cell (CTC) capture device that will provide both enumeration of CTCs and LHRH-R expression on captured CTCs as well as results from AEZS-108 internalization studies that exploit the auto-fluorescence of doxorubicin in captured CTCs. Methods: This is a single-arm, dose-escalation phase I study in men with CRPC to confirm the dose established in a completed phase I trial in females. Eligibility criteria included adequate organ function and progression of disease despite prior therapy with an LHRH agonist and at least one taxane-based regimen. Patients were required to discontinue LHRH agonists to avoid receptor competition. Patients received AEZS-108 every 21 days until progression or unacceptable toxicity for up to 6 cycles. The primary endpoint was safety. Results: Enrollment began in November 2010 and completed in September 2011. Twelve patients were accrued onto 3 dose levels. No DLTs have been noted. At the time of submission, a decrease in PSA was noted in 5 of the 10 evaluable patients. The grade 3 or 4 toxicities were primarily hematologic. Final reports detailing toxicity, RECIST response and PSA response will be reported. All correlative studies will also be reported. Conclusions: AEZS-108 is well tolerated and has demonstrated early signs of antitumor activity in men with CRPC. We will report the recommended dose for the planned phase II study.

Abstract Background: Perifosine, a novel, first-in-class, oral Akt inhibitor, in combination with the antimetabolite Capecitabine is currently in Phase III clinical trials for the treatment of refractory advanced colorectal cancer. We have shown previously that Perifosine in combination with antimetabolites displayed synergistic cytotoxicity and apoptosis in colon cancer and other cell lines [1]. Constitutive NF-kB activation is frequently observed in colorectal cancer and, moreover, is associated with resistance to chemotherapy [2]. Hence we analysed, wether downregulation of the NF-kB pathway contributes to Perifosine mechanism of action and to the observed synergism with antimetabolites in colon cancer cell lines. Methods: Cellular NF-kB activation in nuclear extracts was measured by the TransAm NF-kB p65 ELISA (Active Motif, USA) monitoring binding of NF-kB to its response element. To allow for higher throughput of NF-kB pathway analysis a monoclonal SW620 colon cancer cell line ectopically expressing a NF-kB response element driven reporter gene (Promega, USA) was established. Cellular cytotoxicity and apoptosis assays as well as western blotting were performed according to standard procedures. The analysis of drug combinations was conducted by CalcuSyn software (Biosoft, UK) [3]. Results: Perifosine pretreatment decreased TNFα-induced NF-kB activation in SW620 cells in both independent assay sytems with IC50 values of about 7-10 µM. This effect seems to be specific, since the results were not mimicked by cytotoxic effects of Perifosine. Some of the antimetabolites alone upregulated NF-kB activity in the reporter gene assay. In the presence of these antimetabolites Perifosine was still capable to downregulate NF-kB activity. Conclusions: In summary, these results demonstrate that downregulation of the NF-kB pathway may contribute to the mechanism of action of Perifosine. Moreover, cotreatment with Perifosine has the potential to interfere with the development of antimetabolite resistance due to NF-kB activation in colon cancer. Further experiments addressing Perifosine's mechanism of action in combination with antimetabolites in gastric cancer cell lines are ongoing. References [1] Aicher et al. (2010), Perifosine in combination with antimetabolites induces synergistic effects on cytotoxicity and apoptosis in human colon, multiple myeloma, breast, renal, and liver tumor cell lines. AACR-NCI-EORTC International Conference, Berlin, Germany. [2] Sakamoto and Maeda (2010), Expert. Opin. Ther. Targets 14, 593-601 [3] Chou (2006), Pharmacological Reviews 58, 621-681 Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 2877. doi:1538-7445.AM2012-2877

Antagonisten des growth hormone releasing hormone (GHRH) zeigten eine effektive Tumorwachstumsinhibition bzw. Zellproliferationshemmung bei verschiednen Tumorentitäten, die den hypophysären GHRH-Rezeptor bzw. dessen aktive splice Variante SV1 exprimieren. Rapamycin als mTOR Inhibitor zeigte eine besseres Ansprechen bei hormonrezeptorpositiven metastasierten Mammakarzinompatientinnen in Kombination mit einer endokrinen Therapie gegenüber einer alleinigen endokrinen Therapie. In dieser Studie überprüften wir die Effektivität einer Kombinationstherapie mit einem GHRH Antagonisten in triple negativen Mammakarzinomen.

Aim: To identify the maximum-tolerated dose (MTD) and pharmacokinetics of oral perifosine.Methods: Patients with solid tumours received perifosine at dosages ranging from 100800 mg/week. Eligibility criteria included life expectancy > 12 weeks, WHO performance status <= 2, normal blood, liver and renal functions and no recent anticancer treatment. Drug concentrations were analysed by HPLC-MS/MS.Results: Thirty six patients were recruited (75% males, mean age 54.7 years, performance status 1 in 72.2%). Adverse events included nausea (69.4%), diarrhoea (55.6%), vomiting (52.8%) and abdominal pain (13.9%). Antiemetic regimens including glucocorticoids, dopamine antagonists and 5-HT3-antagonists were used as treatment and/or prophylaxis in 50% of the patients. Though MTD was formally not reached with 800 mg/week, the treatment discontinuation due to diarrhoea and vomiting likely related to perifosine in two cases led to the decision to stop further dose escalation. Pharmacokinetics after a single dose were median t(max) = 8.0-24.2 h, median t(1/2) = 81.0-115.9 h and mean(geo) CL/f = 0.28-0.43 mL/min/kg. Urinary excretion was below 1%. Perifosine slightly accumulated and steady state was nearly reached after 2-3 weeks.Conclusion: Oral perifosine was tolerable up to 600 mg/week in cancer patients when administered with meal and prophylactic antiemetics. Based on its half-life of about 4 days, a weekly regimen may be appropriate. (C) 2010 Elsevier Ltd. All rights reserved.