Drug Delivery to the CNS: Barriers that May Influence Efficacy in Treating Tuberculosis in the Brain

Transcription

1 Drug Delivery to the CNS: Barriers that May Influence Efficacy in Treating Tuberculosis in the Brain TB-Meningitis Blood Brain Barrier astrocyte endfeet TB-Meningitis Rockville MD May 2017 tight junctions capillary lumen endothelial cell layer basement membrane luminal Systemic Pharmacokinetics efflux diffusion influx abluminal William F. Elmquist Pharmaceutics 1

2 Translational research in CNS Drug Delivery - Must keep in mind the big questions! Why Does a Drug Work?? Why Doesn t a Drug Work?? Why Why does this one work, and that one doesn t?? Connect - Disconnect of the PK-PD Relationship Information Pharmacodynamics (events at the target) Balance Flow Pharmacokinetics (conc-time in blood) Information

3 Overview CNS Drug Delivery in the Era of Systems Biology Dose and dosing regimen Traditional PK/PD Black box models the Big Picture Systems model Mechanisms of delivery and action Molecular pharmacokinetics System structure function Nature Biotechnology 23, (2005)

4 Examine drug pharmacokinetics / delivery to CNS sites across several scales Lung Heart Kidney Venous Blood Brain Arterial Blood BBB ECF meso neuronal and glial cell membranes target macro Liver plasma ICF choroid plexus arachnoid membrane ependyma pia mater cerebrospinal fluid Quantitative Qualitative Oscillations in the Thought Cycle micro X Drug

5 Connect - Disconnect of the PK-PD Relationship Understanding Sources of Variability in Drug Response Variability Cycle Genetic Factors - drug targets - drug transporters - drug metabolizing enzymes Environmental Factors - induction - inhibition Physiological Factors - age, disease, etc.

6 Locations of Variability in Drug Efficacy In CNS Tuberculosis Intestinal metabolism Tissue distribution B-TB Target site Oral dosage form Systemic circulation Drug action Intestinal absorption Liver metabolism BBB Cellular delivery TB-Target Presystemic bioavailability questions ( traditional bioavailability) Site-specific bioavailability questions (drug targeting) Targeted Bioavailability 6

7 Locations of Variability in Drug Response Mechanisms that influence the fraction of the drug in the systemic circulation that is available for distribution to target tissue and the exposure of the tissue to the drug Oral - distribution Intestinal of blood flow Tissue Target site dosage - ratio of metabolism total clearance to a distributional distribution clearance formdistributional clearance - membrane permeability, competing carrier-mediated transport (influx Systemic or efflux), protein-binding, intracellular metabolism, tissue circulation transit time, capillary structure Total clearance will affect the availability of the drug in the blood to distribute to the tissue Drug action Intestinal absorption Liver metabolism Systemic clearance Cellular delivery Presystemic bioavailability questions ( traditional bioavailability) Site-specific bioavailability questions (drug targeting) Targeted Bioavailability

8 Physicochemical Properties Physiology / Pathology Drug Metabolism Membrane Permeability Drug Transport Protein Binding Receptor Affinity Gene Regulation Targeted Bioavailability Protein Expression Dosage Regimen Pharmacological / Toxicological Response 8

9 One Location : Blood-brain Barrier Importance of Transporters in the CNS Disposition of Drugs From Lee and Gottesmann. Journal of Clinical Investigation, 1998 illustration by Naba Bora, Medical College of Georgia. 9

10 GLUT1 brain Clin brain P-gp (p-glycoprotein) blood Co-localization GLUT1 - P-gp brain Clout Modifed from: Loscher, Aug Nature Rev. Neurosci. Tight Junction BCRP P-gp 10

11 Therapeutic decisions limited by available data at specific sites input output exchange bolus IC, CED PO, IV plasma brain capillaries extracellular fluid neuronal and glial cell membranes intracellular fluid ICV, IT choroid plexus arachnoid membrane cerebrospinal fluid ependyma pia mater To know, is to measure. Compartmental model for solute exchange in the brain

12 Modeling limited by available mechanistic data at specific sites brain capillaries neuronal and glial cell plasma choroid plexus arachnoid ECF production Surface area Permeability Transporters Metabolism Regional variability extracellular fluid Convection Diffusion Transporters Permeability Metabolism Receptors Regional variability intracellular fluid CSF flow Surface area Permeability Transporters Metabolism Regional variability ependyma pia mater cerebrospinal fluid Convection, Diffusion Permeability, Regional variability

13 Simplified Quantitative Analysis of Drug Transfer In CNS Drug Binding Plasma and Brain Plasma Cu,plasma Cplasma BBB BCSFB PS CL eff CL in [CLin, CLeff, PS] Brain Cbrain Cu,brain CL bulk Ccsf CSF CL meta

14 Simplified Quantitative Analysis of Drug Transfer In CNS Kinetics of distribution - Rate and Extent Rate (onset) - described by maximum concentration (Cmax) Extent (exposure) - described by area under the curve (AUC) Ratio of areas gives tissue partition coefficient Kp = AUC AUC brain plasma Kp, uu = AUC AUC brain unbound plasma unbound Total concs Unbound concs

15 Simplified Quantitative Analysis of Drug Transfer In CNS Extent - partitioning of free concentration K = p, free CL CL in out Sum of clearances in each direction K p, free = PS + CL PS + efflux + CL CL uptake metabolism + CL bulk

16 Simplified Quantitative Analysis of Drug Transfer In CNS Extent - partitioning into a specific brain region Tight-junction opening Substrate for Influx Transporter K p, free = PS + CL PS + efflux + CL CL uptake metabolism + CL bulk Tight-junction opening Substrate for Efflux Transporter Brain Metabolism Fluid flow Active transport CL depends on both capacity and affinity CL Tmax = act Km + C

17 Representative Case Study: The Treatment of Glioblastoma with Inhibition of P53 Degradation MDM2 Inhibitor Minjee Kim, Jann Sarkaria

18 Choice of PDX Glioma Model Apoptosis (%) nm 100 nm Neurosphere Count (%) nm 100 nm GBM10 GBM12 GBM102 GBM108 GBM143 0 GBM10 GBM102 GBM108 GBM143

19 Efficacy of SAR depends on tumor location Heterotopic Xenograft Orthotopic Xenograft Volume (mm 3 ) 2,500 2,000 1,500 1, GBM108 Placebo SAR mg/kg qd n = 7 p = n = Study Day Survival GBM108 P=0.59 n = 12 Placebo SAR mg/kg qd n = Study Day

20 SAR Concentration vs. Time Profiles in Plasma and Brain Plasma Brain Pgp-/- TKO-/- WT, Bcrp-/- Influence of Efflux Transporters at the BBB On Brain Penetration of SAR405838

21 SAR Plasma and Brain Distribution Kinetics Wild-type Mdr1a/1b -/- Bcrp1 -/- Mdr1a/1b -/- Bcrp1 -/- T 1/2 Hr T max Hr C max ng/ml AUC inf_pred hr*ng/ml Vz/F ml/kg CL/F ml/hr/kg AUC brain hr*ng/ml Distribution Advantage

22 orthotopic GBM108 parental line TexasRed Cresyl Violet Blood brain barrier integrity in orthotopic tumors. Near-moribund mice with orthotopic GBM108 tumors were injected with TexasRed-3 kda dextran conjugate 10 min before euthanasia and processed for cresyl violet and fluorescent microscopy on serial histology sections. Accumulation of TRdextran within the tumor reflects disruption of the BBB. Results presented are representative of five mice analyzed. Scale bar = 500 µm. 22

23 Heterogeneous Breakdown of Tumor BBB orthotopic GBM108 parental line 23

24 G108-VEGFA Cell Line Generation Texas-Red Cresyl violet GBM108- VEGFA GBM108-vector

25 A Texas Red Cresyl violet T1 + Gad T2/FLAIR B VEGFA Vector VEGFA VEGFA Vector Vector C H&E Gadavist SAR405838

26 Spatial Distribution of SAR (MALDI-MSI) GBM108-Empty vector GBM108-VEGFA

27 Orthotopic Survival Vector VEGFA Survival Survival day Placebo SAR Days Placebo SAR Days

28 Conclusions for SAR Study SAR405838, a potent MDM2 inhibitor, is subject to BBB efflux This preclinical study indicates enhanced delivery of SAR will improve its efficacy Strategies to overcome limited delivery of drug across BBB will result in better treatment for brain tumors

29 Translation in the Clinic - Delivery and the BBB Deb Brinkmann, Jann Sarkaria (Mayo Clinic)

30 Use of Uptake Transporters in BBB 30

31 Deb Brinkmann, Jann Sarkaria - Mayo Clinic, Rochester Discordance in tumor delineation by 18F-FDOPA PET and MRI. Volumes defined for : A) FDOPA positivity ( ) by PET B) T1 contrast enhancement ( ) on T1 contrast enhanced images C) FLAIR positive (blue) outlined for a single patient 31

32 Structure Structure Volume (cc) from Eclipse PET Volume outside of T1-GAD (cc) from Eclipse PET Volume outside of FLAIR (cc) from Eclipse MR Volume outside of PET (cc) from Eclipse Regions of tumor with intact BBB protected from treatment by efflux transporters and TJ T1-GAD 10.4 N/A N/A 4.9 FLAIR 32.0 N/A N/A 21.7 PET N/A RT_FDOPA02 Grade IV Total Multi-focal FLAIR contour in blue T1-GAD contour in red PET contour in yellow Orthogonal views with crosshairs turned on, for reference

33 Structure Structure Volume (cc) from Eclipse PET Volume outside of T1-GAD (cc) from Eclipse PET Volume outside of FLAIR (cc) from Eclipse MR Volume outside of PET (cc) from Eclipse Regions of tumor with intact BBB protected from treatment by efflux transporters and TJ T1-GAD 47.7 N/A N/A 33.2** FLAIR 54.3 N/A N/A 36.4 PET N/A RT_FDOPA05 Grade IV Total Single FLAIR contour in blue T1-Gad contour in red PET contour in yellow **T1-GAD contour includes post-op cavity (not just enhancement) Orthogonal views with crosshairs turned on, for reference

34 Jann Sarkaria Mayo Clinic Rochester A. B. Screen capture of biopsy planning using the registered 18 F-DOPA PET and T1- CE MRI in the Stealth Neuronavigation System for blue needle locations at: A) a T1 contrast enhancing, PET positive (M+P+) location B) a non-contrast enhancing but PET positive (M-P+) location. 34

35 CT scan T1 CE - MRI T2 - MRI Region specific disease, requires region specific consideration of drug delivery

36 Spatial and Temporal Changes in Delivery of Drugs as Disease Progresses Cu,plasma Plasma Cplasma BBB PS CL eff CL in BCSFB [CLin, CLeff, PS] CT scan T1 CE - MRI T2 - MRI Brain Cbrain Cu,brain CL meta CL bulk Ccsf CSF Questions: 1) In the tuberculomas, is there a change in drug penetration as they encapsulate? 2) Is there active disease in the peripheral regions that show edema? 3) What is the integrity of the BBB at different sites within an infected brain? 4) Are drug concentrations in each region adequate to treat disease? 5) Are there significant differences in delivery limitations amongst drugs used in the necessary combinations in different regions of disease?

37 normal brain 0.29 µg/ml tumor 1.4 µg/ml microdialysis extracellular fluid serum peritumor 10.9 µg/ml 0.32 µg/ml 0.6 µg/ml Rifampicin Spatial Differences in Distribution within the Human Brain

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39 MALDI-MSI for Rifampicin Distribution in the Lung

40 PET scan for distribution of 11C-Rifampicin in M. tuberculosis Infected Mouse Model Liver AUC infected = ng*hr/ml AUC control = ng*hr/ml Blood DeMarco et al AUC infected = ng*hr/ml AUC control = ng*hr/ml Brain AUCinfected = ng*hr/ml Lung AUC infected = ng*hr/ml AUC control = ng*hr/ml AUC control = ng*hr/ml

41 11 C PET RIF

42 Cu,plasma Plasma Cplasma BBB PS CL eff CL in BCSFB [CLin, CLeff, PS] Brain Cbrain Cu,brain CL bulk Ccsf CSF CL meta

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44 Parp1 inhibitor - Talazoparib Talazoparib Pgp substrate - poor penetration into the brain

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46 toxicity potency delivery

47 toxicity delivery potency The sweet spot

48 Delivery across the BBB: Beware of the simplest explanation. Sweet Spot for CNS Rx Empiric Black-Box Pharmacokinetics Systems Pharmacology Intelligent Drug Design Make things as simple as possible, but not simpler. Albert Einstein Mechanistic Molecular Pharmacokinetics