CPRIT Bridging the Gap: Early Translational Research Award RP121060

Transcription

1 CPRIT Bridging the Gap: Early Translational Research Award RP GMP Manufacture, Formulation, and IND directed Toxicology of the Multifunctional Ceramide Catabolism Inhibitor D threo PPMP to enable Phase I Clinical Trials CAS # Barry J. Maurer, MD PhD Cell Biology and Biochemistry, Pediatrics and Medicine TTUHSC SOM Cancer Center Min H. Kang, PharmD Cell Biology and Biochemistry, Pediatrics and Pharmacology TTUHSC SOM Cancer Center C. Patrick Reynolds, MD PhD

2 Conflict of Interest Disclosures The Texas Tech University System (TTUS) and Children s Hospital of Los Angeles (CHLA) holds patents and patent applications (pending) on the use of Fenretinide, Fenretinide-containing drug combinations, and various formulations of Fenretinide and ceramide-modulating co-agents for use in cancer therapy. As an inventor, BJM may potentially benefit financially from the development and future use of this IP through institutional revenue-sharing agreements. BJM is Co-founder, equity holder, and Chief Medical Officer of CerRx, Inc., Lubbock TX, a CPRIT-supported, clinical-stage drug development start-up company that licenses certain IP from CHLA and TTUS. BJM may potentially benefit financially in these capacity.

3 Desirable Properties for a New Chemotherapy novel mechanism p53 independent active in physiological hypoxia complement existing therapy

4 Development of Novel Dihydroceramide Mediated Chemotherapies Fenretinide, 4 HPR Increase ROS in susceptible cancer cells Increase dihydroceramides in susceptible cancer cells (but not in normal cells) Concentration and time dependent manner p53 independent activity retained in hypoxia

5 De Novo Synthetic Pathway of Ceramides 4 HPR Serine + Palmitoyl CoA Serine Palmitoyltransferase HO Fatty Acid 4 HPR OH OH NH 2 D erythro Sphinganine Dihydroceramide Synthases 1 6 Sphingomyelins (non toxic) H O H N O 4 HPR * Dihydroceramides (cytotoxic) HO HN OH O Dihydroceramide Desaturase 1 * Ceramides (cytotoxic) *(variable length) *(variable length) Glucosylceramides (non toxic)

6 Certain Dihydroceramides are Cytotoxic to Cancer Cells Certain but not all dihydroceramides are cytotoxic to cancer cells possibly inacancerorcancertype specific manner. Strong positive correlation between C22:0 and C24:0 dihydroceramide levels and cytotoxicity in ALL T cell leukemia cell lines. C22-DHCer C24-DHCer Cell Line ρ P-value ρ P-value CCRF-CEM 0.74 < <0.001 MOLT < <0.001 COG-LL < <0.001 COG-LL < <0.02 Holliday, Jr., M.W., Cox, S.B., Kang, M.H., and Maurer, B.J., C22:0 and C24:0 dihydroceramides confer cytotoxicity in T cell acute lymphoblastic leukemia cell lines. PLoS ONE, 8(9): e doi: (2013).

7 High Dose Fenretinide Increases Dihydroceramides in Tumor Cell Lines and Xenografts Human Neuroblastoma Tumor Xenografts 10 2 No treatment 4-HPR CHLA No treatment 4-HPR + ketoconazole CHLA 90 pmole DHCer/nmole PO pmole DHCer/nmole PO :0 16:0 18:0 18:1 20:0 20:1 22:0 22:1 24:0 24: :0 16:0 18:0 18:1 20:0 20:1 22:0 22:1 24:0 24:1 DHCer Species DHCer Species 2012 CPRIT Annual Meeting. Abstract #118. Fenretinide/ LYM X SORB TM (4 HPR/LXS) Oral Powder Combined With Ketoconazole, A Fenretinide Metabolism Inhibitor, Synergizes With Vincristine Antitumor Activity Against Recurrent Neuroblastoma Xenografts Lluis A. Lopez Barcons, Hardeep Singh, Barry J. Maurer, Min H. Kang, C. Patrick Reynolds, TTUHSC

8 Can High Dose Fenretinide Be Delivered Clinically? Retinoids are minimally water soluble Previous McNeil/J&J/NCI oral capsule formulation 7 10 M New NCI RAID Supported Formulations with Superior M M

9 Fenretinide/LXS oral powder: Increased CR responses and 3 yr PFS in Relapsed Neuroblastoma in Phase 1 Trial Estimated Probability NANT Overall and Progression-Free Survival (n=34) Excludes Patients with Baseline CR or BM Disease Only Overall Survival Median (95% CI): 30.2 (17.2, 35.6) Months Progression Free Survival Median (95% CI): 4.1 (1.3, 18.8) Months Historical PFS single agents Months Since Treatment Start 2014 ASCO Annual Meeting. Abstract NANT Fenretinide (4 HPR)/Lym X Sorb (LXS) Oral Powder + Ketoconazole in Patients with High Risk (HR) Recurrent or Resistant Neuroblastoma. A New Approaches to Neuroblastoma Therapy (NANT) Consortium Trial. B. J. Maurer, J. Glade Bender, M. H. Kang, J. G. Villablanca, D. Wei, S. Groshen, S. Yang, S. Czarnecki M. Granger, H. Katzenstein, B. Weiss, K.K. Matthay, C. P. Reynolds, and A. Marachelian

10 IV Fenretinide emulsion*: Well tolerated in Phase 1 trials Sustained CR Responses in T Cell Lymphomas Dose Best Diagnosis mg/m 2 /d Response PTCL (AITL) 1810 PRu - (PFS 6 mo) CTCL (Sezary) 1280 CR - (PFS 36+ mo) CTCL 1280 unevaluable - withdrew CTCL 1280/905 SD - much improved skin/pruritis CTCL 905/640 SD - (PFS 3+ mo) PTCL (AITL) 905 CRu - (PFS 30+ mo) - withdrew CTCL 905/640 SD - (2 months) CTCL 905 PR - (PFS 5 mo) CTCL 640 SD - (PFS 8 mo) PTCL (NOS) 640 PD PTCL (AITL) 640 PD Low toxicity profile Transient non symptomatic thrombocytopenia Non symptomatic hypertriglyceridemia *Currently in FDA accelerated approval Phase 2b trial (FEN T14) Relapsed PTCL by CerRx, Inc. Pre-therapy Post -2 cycles 2012 ASCO Annual Meeting Abstract # Phase I trial of fenretinide (4 HPR) intravenous emulsion in hematologic malignancies: A California Cancer Consortium study (PhI 42). Ann Mohrbacher, Min H. Kang, Allen S. Yang, Susan G. Groshen, Lori Vergara, Martin Gutierrez, Anthony J. Murgo, Shivaani Kummar, Donald Quick, C. Patrick Reynolds, Edward M. Newman and Barry J. Maurer.

11 Increasing Fenretinide induced Dihydroceramides Serine + Palmitoyl CoA 4 HPR Serine Palmitoyltransferase HO OH NH 2 D erythro Sphinganine D-t-PPMP Fatty Acid 4 HPR Dihydroceramide Synthases 1 6 Dihydroceramides D-t-PPMP Sphingomyelins (non toxic) 4 HPR Dihydroceramide Desaturase 1 Glucosylceramides (non toxic) Ceramides Guo, W X and Maurer, B. J., unpublished

12 D threo PPMP Increased Fenretinide induced Dihydroceramides and Fenretinide Responses in Cell Line and Xenograft Models (Normalized with total protein) Sphingolipids (fold increase) Control PPMP 4-HPR H+P Survival Fraction 1e+0 1e-1 1e-2 1e-3 SK - N - RA H + t -PPMP H H + e -PPMP 1e HPR, M PPMP, M t - e PPMP PPMP Tumor Size mm^ HPR + D-threo-PPMP improved survivial in KCNR neuroblastoma murine xenograft C1 C Day C3 C4 C5 C6 HP1 HP2 HP3 HP4 HP5 HP6 Survival Fraction H + D-threo SK-N-RA HPR, M D-, or L-threo-PPMP, M H H + L-threo L-threo D-threo

13 CPRIT Bridging the Gap: Early Translational Research Award RP GMP Manufacture, Formulation, and IND directed Toxicology of the Multifunctional Ceramide Catabolism Inhibitor D threo PPMP to enable Phase I Clinical Trials AIM 1. Manufacture1.5 kg of cgmp D threo PPMP using previous GMP process developed via NIH NCI STTR grant. AIM 2. Attempt oral formulation as a powderized, organized lipid matrix (Lym X Sorb (LXS), Avanti Polar Lipids, Alabaster, AL) [else seek intravenous formulation]. AIM 3. Aim 3A) Assay the blood, tissue and tumor levels of 4 HPR + PPMP in mice, assay tumor ceramide levels, verify activity in 4 HPR responsive human xenograft models, and test doxorubicin andpaclitaxelinovariancancerxenograftmodels. AIM 3B) Validate our published HPLC assay for D threo PPMP (murine) in rat and canine;developavalidated tandem mass spectroscopy assay. AIM 4. Subcontract IND directed, rat and canine toxicology studies to support a IND submission for Phase I clinical trials of 4 HPR + PPMP in refractory adult and pediatric malignancies in the Texasbased, South Plains Oncology Consortium. May 2012

14 Battle Plans and Contact With The Enemy Contractor #1 key precursor no longer available in GMP form. Subcontracted custom synthesis required. 15 month delay. Furtheroral formulation work unsuccessful. Subcontracted i.v. formulation development to Contractor #2 (last STTR material) June 2013 Attempted two i.v. formulation routes one proprietary, one suggested by PI. Non proprietary phospholipid emulsion chosen. July 2104 Using engineering batch material and improved detection methodology, Contractor #2 found twounexpected impurities. Impurities traced back tothe newprecursor product. Contractor #1 established and validated improved detection method and GMP material was re purified. September 2014

15 Two Successful Phospholipid Emulsion Formulations Achieved proceeded with non proprietary technology July 2014

16 BEST CLEAN PPMP emulsion and Placebo manufactured. IND directed Toxicology Subcontractor Retained. March 2015

17 IND directed Toxicology Rat only 14 x 5 animal (male) cohorts PK plasma, tissue Clinical Chemistries Necropsy and Histopathology PPMP Bolus dosing: 10 ml/kg 20 ml/kg PPMP 72 hr CIV dosing: 72 hr CIV Controls 15 ml/kg/day Saline 60 ml/kg/day 30 ml/kg/day PPMP Placebo 60 ml/kg/day 60 mg/kg/day 4 HPR Placebo 60 ml/kg/day 4 HPR 72 hr CIV dosing: 4 HPR 30 ml/kg/day PPMP + 4 HPR 72 hr CIV dosing PPMP 30 ml/kg/day + 4 HPR 30 ml/kg/day August 2015

18 SUMMARY IND directed Toxicology PPMPand PPMP+ 4HPR, well tolerated by bolus orciv. Macroscopic observations at necropsyprimarilyassociated with method of administration,(i.e., femoral vein catheter and continuous IV infusion). No significant treatment related histopathologic effects except slight exacerbation in lung inflammation of rats exposed to 4HPR (previously known). The only clinical chemistry parameters altered by any treatment: 1) increased end CIV cholesterol* and bilirubin in higher PPMP containing groups 2) increased end CIV glucose and triglycerides** in (PPMP + 4 HPR). *Phospholipid emulsion? **Soy oil emulsion End October 2015

19 Pharmacokinetics PPMP plasma IV infusion PPMP Dose 30 mg/kg/d 60 mg/kg/d 120 mg/kg/d 60 mg/kg/d + 4 HPR AUC 0?144 (ug hr/ml) 94.7 (8.9) (38.9) (135.8) (136.3) AUMC 0?144 (ug hr 2 /ml) (343.8) (1333.8) (5176.2) (5177.4) MRT (hr) 39.1 (0.1) 38.2 (0.5) 38.0 (0.25) 37.9 (0.2) CL (ml kg 1 /hr 2 ) 13.3 (1.1) 8.1 (1.1) 3.5 (0.32) 4.3 (0.9) Vd (ml kg 1 ) (44.3) (44.6) (12.32) (32.6) Ke (hr 1 ) 0.03(0.00) 0.06 (0.00) 0.02 (0.00) 0.03 (0.00) t 1/2 (hr) 27.1 (0.1) 26.5 (0.4) 26.4 (0.2) 26.3 (0.1) Cmax (micromolar) 2.7 (0.2) 9 (1.0) 42 (3.8) 17.5 (4.0) PENDING PPMP tissues Fenretinide (4 HPR) plasma Fenretinide (4 HPR) tissues November 2015

20 Lessons Learned It can take longer than you think It costs more than it should

21 Acknowledgements Cancer Prevention and Research Institute of Texas (CPRIT) Avanti Polar Lipids, Alabaster AL Stephen W. Burgess, PhD Dennis Graves Dale C. Smith, PhD Particle Sciences, Bethlehem PA Bruce L. Frank, PhD Matthew Bigert Gary Gwozdz Robert Lee, PhD Bioanalytical Sciences Inc. (BASi), Evansville IN L. David Hopper, DVM, PhD, DABT, RQAP GLP Philip A. Downing Kang Lab TTUHSC Hwangeui Cho, PhD Hardeep Singh, PhD Maurer Lab TTUHSC Dong Wang, MD MS Wei Xing Guo, PhD