Andrew H. Rodgers

Simultaneous optimization of pH and micelle concentration in micellar liquid chromatography

A retention model for ionizable compounds in micellar liquid chromatography is derived and verified. The use of the model for the prediction of retention is illustrated and appropriate optimization strategies for the separation of ionizable compounds in Micellar Liquid Chromatography are discussed.

Carbonate and carbamate derivatives of 4-demethylpenclomedine as novel anticancer agents.

PurposeThe purpose of this investigation was to synthesize a series of carbonate and carbamate derivatives of 4-demethylpenclomedine (DM-PEN), the major plasma non-toxic metabolite of penclomedine (PEN) seen in patients. DM-PEN has been observed to be an active antitumor agent in mouse human xenograft tumor models and non-neurotoxic in a rat model, however, activity in intracranially implanted human glioma xenograft models have not been reported. The major goal was to identify derivatives that are active in brain tumors.MethodsDerivatives were prepared from DM-PEN and evaluated in vivo against human U251 glioblastoma, D54 glioblastoma and MX-1 breast tumor xenografts and mammary tumor 16/C that were implanted in the mammary fat pad or intracranially (IC).ResultsCarbonate and carbamate derivatives were found to be superior to DM-PEN against IC growing human glioblastoma xenografts.ConclusionThe activity of the carbonates and carbamates against human tumor xenografts in vivo suggests consideration of these two series of derivatives of DM-PEN for clinical development.

Abstract 1185: A first-in-humans phase I cancer clinical trial for 4-Demthyl-4-cholesteryloxycarbonylpenclomedine (DM-CHOC-PEN).

Purpose: 4-Demethyl-4-cholesteryloxycarbonylpenclomedine (DM-CHOC-PEN) is a poly-chlorinated pyridine cholesteryl carbonate whose MOA is via alkylation of DNA @ N7 - guanine and induces oxidative stress. The main aims of this first-in human trial were to determine maximum-tolerated dose (MTD), safety, dose-limiting toxicities (DLTs) and pharmacokinetics (PK) of DM-CHOC-PEN - IND 68,876. Patients & Methods: DM-CHOC-PEN was administered as a 3-hr IV infusion once every 21-days in 21 patients with melanoma (n=3), colorectal CA (CRC, n=3), and glioblastoma multiforme (GBM) (n= 6); the most frequent diagnoses. The trial allowed enrollment of patients with advanced cancer +/- CNS involvement. The starting dose was 39 mg/m2 with escalations to date up to 98.7 mg/m2. Results: Twenty-one (21) patients have been treated to date. MTD was 2-tiered and defined as 85.8 mg/m2 for patients with liver involvement and, for patients without liver abnormality it is not yet defined but is at least 98.7 mg/m2, for the latter population. The drug was well tolerated with the most common adverse effects being fatigue (n=2), liver dysfunction - elevated bilirubin (Gr-3, n=2; Gr-2, n=1), ALT/AST (Gr-2, n=3), alk phos (Gr-2, n=3) and an allergic reaction (Gr-2, n=1). No neuro/psychological, hematological or renal toxicity observed. Two (2) patients with liver metastasis demonstrated hyperbilirubinemia (Gr-3 SLT) at the 98.7 mg/m2 dose level. Five (5) additional patients with liver disease have been treated at 85.8 mg/m2 level without toxicity - the MTD for that stage of cancer. Dose cohorts @ 98.7 mg/m2 are in progress for non-liver staged patients. PK studies in humans revealed the following profile for DM-CHOC-PEN 70 mg/kg: AUC o-t = 980 mg.h/L, CL - 0.141L/h, T1/2 α - 0.63 h & Tβ - 24.1 h. DM-CHOC PEN and DM-PEN showed a rebound phenomenon @ ∼ 50 hours post-infusion with a T release of 26.7h. Same phenomenon is observed in RBCs (estimation using Monolix 3.2). DM-CHOC-PEN and DM-PEN were detected 3 and 15 days bound to RBCs (after 70 mg/m2); DM-CHOC-PEN was also detected in the urine (Cmax=17.5 μg/mL) until day 15. The AUC was linear for all doses. Similar to the rats, the total lipid profiles in the patients were erratic (2o to the lipid emulsion vehicle) during the 3-h infusion period, and then returned to pre-treatment values after 24 h. The triglycerides were the most significantly affected. DM-CHOC-PEN could be identified in spinal sarcoma tissue (in 190 ng/g quantities) obtained surgically from a patient 21- days post single injection of 39 mg/m2. Conclusion: DM-CHOC-PEN is safe at the presented dose levels and shows a favorable PK profile. Seven (7) patients have observed responses or significant PFS, including 5 with CNS involvement. DM-CHOC-PEN is well tolerated with manageable toxicities. Complete patient responses/toxicities will be presented. Supported by NCI/SBIR grant -1R43CA132257. Citation Format: Roy S. Weiner, Philip Friedlander, Craig Gordon, Yvonne Saenger, Marcus Ware, Tallat Mahmood, AH Rodgers, Gerard Bastian, S Urien, Lee Roy Morgan. A first-in-humans phase I cancer clinical trial for 4-Demthyl-4-cholesteryloxycarbonylpenclomedine (DM-CHOC-PEN). [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 1185. doi:10.1158/1538-7445.AM2013-1185

Abstract A086: Early clinical trial results for 4-demethyl-4-cholesteryloxycarbonylpenclomedine (DM-CHOC-PEN) in adolescents and young adults (AYA) with cancers

Background: 4-Demethyl-4-cholesteryloxycarbonylpenclomedine (DM-CHOC-PEN) is a poly-chlorinated pyridine cholesteryl carbonate with a MOA via bis-alkylation of DNA @ N7-guanine and N4-cytosine that has completed phase I and II trials in adult subjects with cancer that had +/- CNS involvement (AACR #1185, 2013; AACR #CT 129, 2017). The aims were to assess clinical responses, monitor toxicities/safety, and confirm MTDs for IV administered DM-CHOC-PEN (IND 68,876). We report here responses and toxicities seen in a phase I DM-CHOC-PEN trial with AYA subjects who have cancer (some of whom had CNS involvement). Subjects and Methods: DM-CHOC-PEN was administered as a 3-hr IV infusion once every 21 days to subjects with advanced cancer in escalating doses from 39 - 98.7 mg/m2. The dosing schedule accounted for the liver toxicities seen in the adult phase I trial and was 2-tiered: for subjects with liver involvement, the cutoff was 85.8 mg/m2 and for subjects with normal liver function was 98.7 mg/m2. Results: Twelve (12) AYA subjects have been treated to date (with or without CNS involvement). The common tumor types treated were oligoastrocytoma, astrocytoma, GBM; leukemia (ALL), lymphoma (NHL), melanoma, breast, and lung cancers (NSCL). Most subjects (9) had CNS involvement. The drug was well tolerated; the most common adverse effects were fatigue (17%). No neuro/cognitive, liver dysfunction, hematologic, cardiac, renal, or GI toxicities were observed. PK modeling revealed that AUCs were parallel for all dose levels (39-98.7 mg/m2). The Cmax for DM-CHOC-PEN and DM-PEN (4-demethylpenclomedine, a metabolite) were 3 and 24 hours, respectively for the AYA subjects, similar to what was seen for older adults. However, the drug and metabolite were still detectable in plasma and rbcs for 21 to 50 days in the AYA (15 - 39 yo) group vs. 3 to 60 yo) subjects (AACR #1185, 2013). Differences in PK profiles between AYA and older adult subjects will be reviewed in depth. Three (3) of the AYA subjects (1 each with NSCLC, ALL, and astrocytoma involving the CNS) have responded with CR/PR (RECIST 1.1), improved QOL/PFS (Kaplan-Meier), and OS from 8 to 26+ mos. All subjects will be reviewed. Conclusion: DM-CHOC-PEN is safe in doses of 39 - 98.7 mg/m2 and has produced objective responses with manageable toxicities in AYA subjects with cancer involving the CNS. Complete data on subject responses and observed toxicities will be presented. We propose a 3-stage mechanism for drug entry into the CNS and into cancer cells via reversible binding with RBCs and then association with L-glutamine transport into cells. Supported by NCI/SBIR grants - R43/44CA132257 and R43CA203351A and NIH NIGMS 1 U54 GM104940. Citation Format: Lee Roy Morgan, Andrew H. Rodgers, Roy S. Weiner, Tallat Mahmood, Marcus L. Ware, Manish Bhandari, Philip Friedlander. Early clinical trial results for 4-demethyl-4-cholesteryloxycarbonylpenclomedine (DM-CHOC-PEN) in adolescents and young adults (AYA) with cancers [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2017 Oct 26-30; Philadelphia, PA. Philadelphia (PA): AACR; Mol Cancer Ther 2018;17(1 Suppl):Abstract nr A086.

Comparative Anticancer Activity in Human Tumor Xenograft Models, Preclinical Pharmacology and Toxicology for 4- Hydroperoxyifosfamide (HOOI): A Potential Neuro-Alkylating Agent for Primary and Metastatic Cancers Involving the Central Nervous System

Background: 4-Hydropeoxyifosfamide (HOOI) is a hydroperoxy derivative of ifosfamide that was developed as an anticancer agent that can penetrate the blood-brain barrier (BBB), which can be potentially useful in the management of brain tumors. Methods: A novel synthetic scheme for HOOI is presented and verified. HOOI and an HOOI L-lysine salt were prepared and mice implanted intracranially (IC) and in the mammary fat pad with human U251 glioblastoma, D54 glioblastoma, and MX-1 breast tumor xenografts and treated with HOOI IP once daily for 1–5 days. The animals were monitored for responses, increased long-term survival (ILS) and long-term survival (LTS). Mice, rats, and dogs received single IV doses of HOOI in a wide range of concentrations and results are compared and presented herein. Results: HOOI has been synthesized as per a new route in 67% yield. The drug is stable when frozen in the absence of moisture; however, as a lysine salt the drug is stable in solution and as a lyophilized product. HOOI produced complete responses with improved long-term survival against IC implanted U251 glioblastoma, D54 glioblastoma, and MX-1 breast tumor xenografts in mice. The drug was superior to 4-demethyl-4-cholesteryloxycarbonylpenclomedine (DM-CHC-PEN) and BCNU vs. IC implanted tumor models. The HOOI lysine salt demonstrated equal activity to that of HOOI alone. Over

Abstract CT052: Clinical trial results for 4-demethyl-4-cholesteryloxycarbonylpenclomedine (DM-CHOC-PEN) in cancers involving the CNS

Background: 4-Demethyl-4-cholesteryloxycarbonylpenclomedine (DM-CHOC-PEN) is a poly-chlorinated pyridine cholesteryl carbonate with a MOA via bis-alkylation of DNA @ N7-guanine and N4-cytosine that has completed Phase I and II trials (the latter on subjects with CNS involvement) [AACR #1185, 2013; AACR #CT 129]. The primary aim was to assess clinical response and secondary aims to monitor toxicities/safety and verify the MTDs for IV administered DM-CHOC-PEN that derived in Phase I study (IND 68,876). We report here the responses and toxicities seen in all the subjects treated. Subjects & Methods: In Phase I, DM-CHOC-PEN was administered as a 3-hr IV infusion once every 21 days to subjects with advanced cancer; cohorts received escalating doses from 39 - 111 mg/m2. The Phase II dose schedule was 2-tiered: 85.8 mg/m2 for subjects with liver involvement and 98.7 mg/m2 for subjects with normal livers. Results: Fifty two (52) subjects have been treated to date - 25 in Phase I (cancer subjects with or without CNS involvement) and 27 in Phase II (with CNS involvement). The common tumor types treated were primary brain cancers and melanoma, breast, and lung cancers involving the CNS. The drug was well tolerated; the most common adverse effects were fatigue (17%), reversible liver dysfunction (9%) and nausea (11%). No neuro/psychological, hematological, cardiac or renal toxicities were observed. PK modeling revealed that AUCs were parallel for all dose levels (39-111 mg/m2). The Cmax for DM-CHOC-PEN and DM-PEN (4-demethylpenclomedine, a metabolite) were 3 and 24 hours, respectively. Both DM-CHOC-PEN and DM-PEN were detected 3 to 15 days after administration associated (up to 50%) with rbcs. Of interest, young adults ( Citation Format: Roy S. Weiner, Lee Roy Morgan, T Mahmood, R. Kawauchi, C. Gordon, ML Ware, M. Matrana, TM Cosgriff, AH Rodgers, G. Bastian, M. Bhandari, J-J Zou. Clinical trial results for 4-demethyl-4-cholesteryloxycarbonylpenclomedine (DM-CHOC-PEN) in cancers involving the CNS [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr CT052. doi:10.1158/1538-7445.AM2017-CT052

Abstract CT129: Phase II clinical trial results for 4-demethyl-4-cholesteryloxycarbonylpenclomedine (DM-CHOC-PEN) in NSCLC involving the CNS

Background: 4-Demethyl-4-cholesteryloxycarbonylpenclomedine (DM-CHOC-PEN) is a poly-chlorinated pyridine cholesteryl carbonate with a MOA via bis-alkylation of DNA @ N7-guanine and N4-cytosine that has completed a Phase I study [AACR #1185, 2013] and is being evaluated in a Phase II trial in patients with primary brain cancers and with melanoma, breast, and lung cancers with metastases to brain. The aims are to assess clinical response when DM-CHOC-PEN is administered I.V. at MTD and to monitor duration of responses and safety (IND 68,876). We report here the responses and toxicities seen in patients with NSCLC involving the CNS. Patients & Methods: In Phase I, DM-CHOC-PEN was administered as a 3-hr IV infusion once every 21 days to patients with advanced cancer; cohorts received escalating doses from 39 - 111 mg/m2. The Phase II dose schedule is 2-tiered: 85.8 mg/m2 for patients with liver involvement and 98.7 mg/m2 for patients with normal livers. Results: Fifty two (52) patients have been treated to date - 26 in Phase I (cancer patients with or without CNS involvement) and 26 in Phase II (with CNS involvement). The drug was well tolerated; the most common adverse effects were fatigue (17%), reversible liver dysfunction (9%) and nausea (11%). No neuro/psychological, hematological, cardiac or renal toxicities were observed. PK modeling revealed that AUCs were parallel for all dose levels (39-111 mg/m2). The Cmax for DM-CHOC-PEN and DM-PEN (4-demethylpenclomedine, a metabolite) were 3 and 24 hours, respectively. Both DM-CHOC-PEN and DM-PEN were detected 3 to 15 days after administration associated (up to 50%) with rbcs. DM-CHOC-PEN was also detected in CNS tumor tissue obtained surgically from five (5) patients - concentrations of 75-210 ng/g, 22 days to 9 mos. post treatments at doses of 39 or 98.7 mg/m2 of drug. To date, 16 patients with lung cancer (11 with NSCLC involving the CNS) have been treated. Seven of the 11 patients with NSCLC involving the CNS (incl. 6 with cerebellar disease) have responded with CR/PR (RECIST 1.1) and improved OS/QOL/PFS (Kaplan-Meier) lasting 6+ - 21+ mos. Conclusion: DM-CHOC-PEN is safe at these dose levels and has produced objective responses with manageable toxicities in NSCLC involving the CNS. Complete data on patient responses and observed toxicities will be presented. We propose a 2-stage mechanism for drug entry into the CNS and into NSCLC cells via reversible binding with RBCs and then L-glutamine transport into cells. Supported by NCI/SBIR grants - R43/44CA132257 and NIH NIGMS 1 U54 GM104940 - the latter funds the Louisiana Clinical and Translational Science Center. Citation Format: Roy S. Weiner, T Mahmood, C Gordon, ML Ware, LR Morgan, TM Cosgriff, AH Rodgers, G Bastian, R Kawauchi, M Matrana, J-J Zou. Phase II clinical trial results for 4-demethyl-4-cholesteryloxycarbonylpenclomedine (DM-CHOC-PEN) in NSCLC involving the CNS. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr CT129.

Abstract C152: 4-Demethyl-4-cholesteryloxycarbonylpenclomedine (DM-CHOC-PEN) as a cytotoxic L-glutamine agonist in non-small cell lung cancer (NSCLC) involving the CNS

Purpose: 4-Demethyl-4-cholesteryloxycarbonylpenclomedine (DM-CHOC-PEN) is a poly-chlorinated pyridine cholesteryl carbonate whose MOA is via bis-alkylation of DNA @ N7 - guanine and N6 - cytosine. DM-CHOC-PEN has undergone a Phase I study (allowed enrollment of subjects with advanced cancer +/- CNS involvement) and is being evaluated in a Phase II trial in subjects with advanced cancer involving the brain. Impressive objective responses and improved PFS/overall survival have been observed in subjects with NSCLC involving the CNS. The main aims of this presentation are to document a mechanism for DM-CHOC-PEN9s penetration into NSCLC metastases involving the CNS via L-glutamine (GLM) transport. Methods: Human cancer cell lines [NSCLC - H2087 & H460 and SCLC - SHP-77 & H1417 - obtained from ATCC] were maintained in culture with complete RPMI (5% FBS, pen/strep/Amp. B), supplemented w/ L-glutamine [300 mg/500 mL medium, Sigma] @ 37oC and in moist 5% CO2 conditions. DM-CHOC-PEN was dissolved in tetrahydrofuran, impregnated into ChemoChads® that deliver drug 0.25 - 5 μg/mL in culture. Cells were grown in culture w/wo supplemental GLM for 36 h and then ChemoChads were added. Cytotoxicity was conducted in multi-well plates using the Cytotec® Assay and measuring IC50 values after 24 hr. All assays were conducted in triplicate. Results: All 4-cell lines grew well in complete RPMI supplemented with L-glutamine. Both SCLC cell lines grew well as clumps of small single cells in suspension, +/- GLM suppl. but were not sensitive to DM-CHOC-PEN, w/ or wo/ GLM supplements. In contrast, both NSCLC cell lines grew as coalescing islands of adhered well-differentiated cells in GLM supplemented medium, but did not grow well in GLM-free medium. Of interest, both NSCLC cell lines were sensitive to DM-CHOC-PEN (IC50 0.25-0.75 μg/mL) w/ GLM-suppl. medium, but were not sensitive (IC50 >5 μg/mL) in GLM-free medium. However, when GLM was added to the GLM-free medium w/ DM-CHOC-PEN, the cells resumed sensitivity to the drug. Similar findings have been observed for 6/7-human NSCLC explants obtained from CNS surgical samples (which will be discussed). Conclusion: The common structural similarities between DM-CHOC-PEN, GLM and Ɣ-aminobutyric acid (GABA) could allow the sharing of a transport/receptor mechanism into the cerebellum and other CNS safe havens that provide a favorable microenvironment for NSCLC cells to colonize, thrive and grow. The drug when administered IV associates with erythrocytes (∼50%), which facilitates its entry into the neoplastic cerebral circulation and now support for its transport into metastatic NSCLC involving the CNS via the GLM transport system is presented. In the absence of GLM, the GLM-transport system shuts down. Of clinical interest is that all subjects with cerebellar NSCLC lesions have responded to DM-CHOC-PEN in the clinical trials with objective responses, improved PFS (6-17+ months) and improved QOL (AACR, Abst. 1161, 2015). The cerebellum offers a unique microenvironment rich in chemical transmitters and amino acids - in particular GLM and Ɣ-aminobutyric acid (GABA). Supported by NCI/SBIR grant - R43/44CA132257. Citation Format: Lee Roy Morgan, Jr., Ed Benes, Andrew H. Rodgers, Tallat Mahmood, Roy S. Weiner, Marcus L. Ware. 4-Demethyl-4-cholesteryloxycarbonylpenclomedine (DM-CHOC-PEN) as a cytotoxic L-glutamine agonist in non-small cell lung cancer (NSCLC) involving the CNS. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2015 Nov 5-9; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(12 Suppl 2):Abstract nr C152.

Abstract CT218: Results from the early cancer clinical trials for 4-demethyl-4-cholesteryloxycarbonylpenclomedine (DM-CHOC-PEN)

Proceedings: AACR 106th Annual Meeting 2015; April 18-22, 2015; Philadelphia, PA Purpose: 4-Demethyl-4-cholesteryloxycarbonylpenclomedine (DM-CHOC-PEN), is a poly-chlorinated pyridine cholesteryl carbonate whose MOA is via alkylation of DNA @ N7 - guanine and N6 - cytosine and via oxidative stress. DM-CHOC-PEN underwent a phase I study in patients with advanced cancer +/- CNS involvement and is being evaluated in a phase II trial in patients with primary brain cancer and brain metastases from melanoma, breast, and lung cancers. The aims are to assess clinical responses when DM-CHOC-PEN is administered I.V, at maximum tolerated dose (MTD) and to monitor safety/toxicities, pharmacokinetics, and cardiac functions - IND 68,876. Patients & Methods: In phase I, DM-CHOC-PEN was administered as a 3-hr IV infusion once every 21-days to patients with advanced cancer - melanoma (n = 3), colorectal CA (n = 4), breast (n = 3), lung (n = 8) and glioblastoma multiforme (GBM) (n = 9) - the most common tumor treated. Cohorts were treated with escalating doses from 39 to 111 mg/m2. The phase II dose schedule is 2-tiered: 85.8 mg/m2 for patients with liver involvement and 98.7 mg/m2 for patients with normal livers. Results: Forty (40) patients have been treated to date - 27 in phase I and 13 in phase II. The drug was well tolerated; the most common adverse effects were fatigue (n = 2), liver dysfunction - elevated bilirubin (Gr-3, n = 3; Gr-2, n = 1), ALT/AST (Gr-2, n = 3), alk phos (Gr-2, n = 3), nausea (Gr-1/2, n = 5) and an allergic reaction (Gr-2, n = 1). Three (3) patients with liver metastasis had hyperbilirubinemia (Gr-3 SLT) - two (2) at 98.7 mg/m2 and one (1) at 111 mg/m2 levels. No neuro/psychological, hematological, cardiac or renal toxicities were observed. PK studies revealed the following profile for DM-CHOC-PEN 98.7 mg/m2: AUC o-t = 1850 mg.h/L, CL - 3.0 L/h, T1/2 α - 3.3 h & Tβ - 79.1 h. DM-CHOC PEN and DM-PEN (metabolite) showed a rebound phenomenon at ∼50 hours post-infusion with a T release of 26.7 h for plasma and rbcs. DM-CHOC-PEN and DM-PEN were detected 3 and 15 days bound to RBCs (70 - 111 mg/m2); DM-CHOC-PEN was also detected in the urine (Cmax = 17.5 μg/mL) until day 21. The AUC was linear for all doses. DM-CHOC-PEN was detected in spinal sarcoma and in lung cancer tissues (75 & 190 ng/g, resp.) surgically obtained from patients 21-days post single injection of 39 & 98.7 mg/m2, resp. Patients receiving dexamethasone demonstrated lower blood levels of DM-CHOC-PEN along with induction of steroid esterase activities. After multiple doses, DM-CHOC-PEN also induced steroid esterase levels, which reversed within 4 weeks. Steroid esterase assays may be a valuable companion assay. Conclusion: DM-CHOC-PEN is safe at the presented dose levels and shows a favorable PK profile. To date, 15 patients have had responses with significant PFS/OS, including 10 with CNS involvement. DM-CHOC-PEN is well tolerated with manageable toxicities. Complete patient responses/toxicities will be presented. Supported by NCI/SBIR grant - R43/44CA132257 Citation Format: Roy S. Weiner, P Friedlander, T Mahmood, Adilia Hormigo, C Gordon, Y Saenger, ML Ware, VK Thirukonda, VM Patel, TJ Cosgriff, AH Rodgers, LR Morgan, G Bastian. Results from the early cancer clinical trials for 4-demethyl-4-cholesteryloxycarbonylpenclomedine (DM-CHOC-PEN). [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr CT218. doi:10.1158/1538-7445.AM2015-CT218

Abstract A264: Response of CNS melanoma to 4-demethyl-4-cholestrylcarbonylpenclomedine (DM-CHOC-PEN) and cell death.

Introduction: DM-CHOC-PEN is a polychlorinated pyridine cholesteryl carbonate, which has completed a Phase I clinical trial in patients with advanced cancer - IND 68,876 (AACR 1185, 2013). DM-CHOC-PEN produced responses with improved cerebral progression free intervals (CPFI) in patients with CNS melanoma, breast, and lung cancers. CNS melanoma is of especial interest since there is no acceptable therapy for this stage of disease. DM-CHOC-PEN9s principle MOA is via alkylation of DNA with N7-guanine, however, in melanoma, there is also generation of reactive oxygen species (ROS) via the DOPA/melanin pathway resulting in melanin encapsulation of melanoma cells and cell death. In mice the drug accumulated in intracerebrally/cerebellar implanted tumors, and not in normal CNS tissue, with no Purkinje cell degeneration; in contrast to drugs such as penclomedine, phencyclidine, ibogaine, etc. that also accumulate in the cerebellum but with cellular destruction. Patients with metastatic CNS cancer treated with DM-CHOC-PEN demonstrated some euphoria, but no CNS toxicity. Methods: B-16 melanoma cells were cultured in RPMI media with 5% FBS and pen/strep at 37oC in a CO2 incubator. Drugs were added to the cells during growth phase and removed after 16 hrs; responses were monitored. Adult female C57BL mice in groups of 5-6 with growing B-16 melanoma were dosed IP daily (75-200 mg/kg) for 5 days with DM-CHOC-PEN or saline and monitored daily until death or moribund and sacrificed. Animals were sacrificed, brain tumor and normal tissue removed and prepared for histology exams. Patients in the Phase I trial (DTI-021) with CNS melanoma were treated with 86.5 mg/m2 IV q 21 days and CNS lesions were monitored with MRI during the Phase I clinical trial (DTI-021) [AACR 1185, 2013]. Results: In vitro, DM-CHOC-PEN had an IC50 0.5 µg/mL against B-16 melanoma cells. Mice bearing B-16 melanoma treated with DM-CHOC-PEN (200 mg/kg/d x 5d, IP) alone vs. saline controls demonstrated %ILS of 142% - supporting the in vitro observations. Patients with CNS melanoma treated with DM-CHOC-PEN demonstrated objective responses and improved CPFI with associated melanin deposits similar to those seen with mice. DM-CHOC-PEN induced ROS via DOPA/DOPA oxidase-melanin deposits and cell death in mice with melanoma. Discussions - DM-CHOC-PEN bound to erythrocytes crosses the BBB and accumulates in CNS melanoma (not normal tissue) with intracellular DOPA-DOPA quinone and melanin formation that interrupts cancer cell metabolism and cellular death via ROS. The drug is not recycled into the systemic circulation. Electronic modeling studies support DM-CHOC-PEN9s ability to act as a pyridinium co-factor in the transfer of electrons from DOPA to cytochrome c and the intermediary metabolism pool with generation of ROS. Oxidative stress and ROS generation from a DM-CHOC-PEN/DOPA-melanin pathway has now been confirmed. Cerebellar lesions appear to be especially sensitive. Conclusion: Responses and CPFI for patients with CNS cancers (and no CNS toxicity) seen in the Phase I study have been reported [AACR 1185, 2013]. The principle human toxicity was hepatic. The role for the DOPA/DOPA quinone-melanin pathway with ROS formation will be presented as a key MOA for the drug, with emphasis on cerebellar melanoma lesions. Supported in part by: NCI/ SBIR grants R43/44CA85021 and R43/44CA132257. Citation Information: Mol Cancer Ther 2013;12(11 Suppl):A264. Citation Format: Lee Roy Morgan, Edmund Benes, Andrew H. Rodgers, Roy S. Weiner, Marcus L. Ware, Philip Friedlander, Yvonne Saenger, Craig Gordon, Tallat Mahmood, Branko Jursic, Gerard Bastian. Response of CNS melanoma to 4-demethyl-4-cholestrylcarbonylpenclomedine (DM-CHOC-PEN) and cell death. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr A264.