SBIR-STTR Award

Chemotherapy-induced peripheral neuropathy (CIPN) can be a chronic, severely debilitating consequence of cancer therapy for which there are no effective management strategies. Moreover, upwards of 80% of CIPN patients reported using prescription opioids for pain management despite the fact that there is only weak evidence that long-term continuation of opioids provides clinically significant pain relief in these patients. Mitochondrial dysfunction, oxidative stress, and inflammation have all been implicated in its etiology. We have shown that the non-psychoactive cannabinoid cannabidiol (CBD) prevents the development of CIPN in a mouse model of paclitaxel-induced cold and mechanical allodynia. In vitro, we observe that paclitaxel increases microglial expression of several putative mediators of neuropathic pain, and that this effect can be blocked by CBD in a mitochondrial Na+/Ca2+ exchanger (mNCX)- dependent manner. We have also recently shown that a more potent, hydrophilic analogue of CBD, KLS-13019, protects against paclitaxel-induced oxidative stress in cultured dorsal root ganglia neurons, and that the mechanism underlying this neuroprotection is also regulation of intracellular calcium via the mNCX. Preliminary results demonstrate that KLS-13019 can attenuate mechanical sensitivity associated with CIPN while also reducing microglial activation and T cell infiltration into the spinal cord. Our central hypothesis is that administration of CBD or KLS-13019 helps preserve Ca2+ homeostasis by promoting activity of the mNCX, which in turn protects from both mitochondrial dysfunction and microglial activation to prevent the neuronal and glial changes associated with the development and maintenance of paclitaxel-induced neuropathic pain. Results from experiments in AIM 1 will demonstrate that the neuroprotective properties of CBD and KLS-13019 can be reduced by pharmacological or gene knockdown of the mNCX in a statistically significant manner. Results from experiments in AIM 2 will further confirm the i.p. and p.o. efficacy of KLS-13019 vs CBD to prevent or reverse mechanical sensitivity and neuroinflammation in a mouse model of paclitaxel-induced neuropathic pain and that repeated administration of these compounds does not lead to analgesic tolerance. Remarkably, the non- psychoactive CBD has also been shown to inhibit cue-induced heroin-seeking and neurochemical correlates thereof in a rat model of relapse and decrease heroin craving in a small human study. Experiments in AIM 3 are designed to test the hypothesis that KLS-13019 and CBD will attenuate reinstatement of morphine seeking behavior in a rat model of opioid relapse. The overall impact of the results from the proposed research will be significant advancements into 1) identification of specific mechanisms that induce CIPN, 2) application of this knowledge to facilitate design of novel treatment strategies for neuropathic pain, and 3) novel treatment strategies to reduce or replace prescription opioid use and decrease prescription opioid abuse.

Public Health Relevance Statement:
Chemotherapy-induced peripheral neuropathy (CIPN) can be a chronic, severely debilitating consequence of cancer therapy for which there are no effective management strategies. Moreover, upwards of 80% of CIPN patients reported using prescription opioids for pain management, despite the weak evidence of their efficacy and the risks of long term dependence. Mitochondrial dysfunction, calcium dysregulation, oxidative stress, and inflammation have all been implicated in its etiology. Cannabidiol (CBD) is a non-psychoactive component of Cannabis sativa is effective in both treating CIPN and relieving opiate dependence. However, CBD has severe limitations in terms of potency, safety, oral bioavailability, and regulatory restrictions. Kannalife has solved these problems in its patented series of derivatives that include KLS-13019. This grant will demonstrate the efficacy of KLS- 13019 in models of CIPN and opiate dependence, and will further elucidate its mechanism of action in regulation of calcium levels and inflammatory sequelae.

Project Terms:
Pain management; pain treatment; Pain Therapy; Pain Control; Analgesics; painkiller; pain reliever; pain medication; pain killer; Antinociceptive Drugs; Antinociceptive Agents; Anodynes; Analgesic Preparation; Analgesic Drugs; Analgesic Agents; Opioid Analgesics; opioid painkiller; opioid pain reliever; opioid pain medication; opioid anesthetic; opioid analgesia; opiate pain reliever; opiate pain medication; opiate analgesic; opiate analgesia; inhibitor/antagonist; inhibitor; Architecture; Engineering / Architecture; Back; Dorsum; Behavior; Biological Availability; Physiologic Availability; Biologic Availability; Bioavailability; Calcium; Factor IV; Coagulation Factor IV; Ca++ element; Blood Coagulation Factor IV; Cannabidiol; Cannabinoids; cannabinoidergic; cannabinergic; Pharmaceutical Chemistry; Pharmaceutic Chemistry; Medicinal Chemistry; Cues; Heroin; Diamorphine; Diacetylmorphine; Pharmacotherapy; drug treatment; Drug Therapy; Pharmaceutical Preparations; drug/agent; Pharmaceutic Preparations; Medication; Drugs; Exhibits; Spinal Ganglia; dorsal root ganglion; Dorsal Root Ganglia; Genes; Goals; Grant; Hippocampus (Brain); hippocampal; Hippocampus; Cornu Ammonis; Ammon Horn; Homeostasis; Physiological Homeostasis; Autoregulation; Human; Modern Man; Hydrogen Bonding; H-bond; In Vitro; Inflammation; Lead; heavy metal lead; heavy metal Pb; Pb element; Lymphocyte; lymph cell; Lymphocytic; Maintenance; Mitochondria; mitochondrial; Morphine; Statex SR; Roxanol; Oramorph SR; Oramorph; Morphia; MSir; MS Contin; Kadian; Infumorph; neurochemistry; neurochemical; Neurons; neuronal; Neurocyte; Neural Cell; Nerve Unit; Nerve Cells; opioid abuse; opioid drug abuse; opiate drug abuse; opiate abuse; Legal patent; Patents; Patients; Peripheral Nervous System Diseases; Peripheral Neuropathy; Peripheral Nervous System Disorders; Peripheral Nerve Diseases; PNS Diseases; Permeability; Drug Kinetics; Pharmacokinetics; Pharmacology; Problem Solving; Rattus; Rats Mammals; Rat; Common Rat Strains; Relapse; Research; Risk; Safety; Self Administration; Solubility; Spinal Cord; Medulla Spinalis; T-Lymphocyte; thymus derived lymphocyte; Thymus-Dependent Lymphocytes; T-Cells; Testing; Mediating; Brain-Derived Neurotrophic Factor; BDNF; Paclitaxel; Taxol Konzentrat; Taxol A; Taxol; Praxel; Paclitaxel (Taxol); Bristaxol; Asotax; Anzatax; Pro-Oxidants; Oxygen Radicals; Active Oxygen; Reactive Oxygen Species; improved; Area; Surface; Chronic; Phase; Series; Opiates; Opioid; Oxidative Stress; Neuroprotective Drugs; Neuroprotectants; Neuroprotective Agents; analog; chemoattractant cytokine; SIS cytokines; Intercrines; Homologous Chemotactic Cytokines; Chemotactic Cytokines; chemokine; Attenuated; Infiltration; Inflammatory; Intravenous; Knowledge; Mechanics; mechanical; Dependence; Side; Oral; Opiate Addiction; opioid dependent; opioid dependence; opioid addiction; Opiate Dependence; success; alkalinity; basicity; hydrophilicity; lipophilicity; neuroprotection; Animal Model; model organism; model of animal; Animal Models and Related Studies; attenuation; aqueous; Pharmacology and Toxicology; novel; Prevention; Cannabis sativa plant; Cannabis sativa; C. sativa; C sativa; Reporting; chemotherapeutic agent; Regulation; Modeling; craving; Property; cancer therapy; anticancer therapy; anti-cancer therapy; Malignant Neoplasm Treatment; Malignant Neoplasm Therapy; Cancer Treatment; Bioavailable; prevent; preventing; Etiology; disease causation; causation; Causality; Acidity; Mediator of activation protein; Mediator of Activation; Mediator; Drug Addiction; Drug Dependency; Drug Dependence; Chemical Dependence; Cytokine Suppression; Applications Grants; Grant Proposals; in vitro Assay; in vivo; anti-inflammatory effect; Antiinflammatory Effect; STTR; Small Business Technology Transfer Research; Process; developmental; Development; neuropathic pain; painful neuropathy; knockdown; knock-down; neuroinflammatory; neuroinflammation; designing; design; neural dysfunction; Neuronal Dysfunction; excitotoxicity; therapy efficacy; therapeutically effective; therapeutic efficacy; intervention efficacy; Treatment Efficacy; human study; migration; innovative; innovate; innovation; clinical significance; clinically significant; mitochondrial dysfunction; murine model; mouse model; treatment strategy; effective treatment; effective therapy; biomarker; biologic marker; bio-markers; Biological Markers; mechanical allodynia; compare efficacy; comparative efficacy; prescription opiate abuse; opioid prescription drug abuse; opioid medication abuse; prescription opioid abuse; prescription opiate; prescribed opioid; prescribed opiate; opioid medication; opiate medication; licit opioid; prescription opioid; phytocannabinoid; Chemotherapy-induced peripheral neuropathy; opioid intake; opioid drug use; opioid consumption; opiate use; opiate intake; opiate drug use; opiate consumption; opioid use; experimental research; experiment; experimental study; chronic pain disorder; chronic pain condition; chronic painful condition; relieve pain; pain relief

Neuropathic pain remains a challenging neurologic disorder that adversely affects quality of life and presents a large unmet medical need. Chemotherapy-induced peripheral neuropathy (CIPN) is a chronic, severely debilitating consequence of cancer therapy for which there are no effective management strategies. Upwards of 80-97% of CIPN patients reported using prescription opioids for this pain management. Mitochondrial dysfunction, oxidative stress, and inflammation have all been implicated in CIPN etiology. In a mouse model of paclitaxel-induced pain sensitivity, we have previously reported that cannabidiol (CBD) is effective in preventing the onset of this treatment consequence. Now a new CBD analogue (KLS-13019) has been discovered in our laboratory that has improved drug-like properties in comparison to CBD, while retaining neuroprotective properties. In our Phase 1 STTR, the previous neuroprotective effects of CBD to prevent the development of mechanical sensitivity in the presence of paclitaxel were confirmed and extended to the structural analogue KLS- 13019. Both compounds were equi-effective and equi-potent following oral administration. In the reversal studies, CBD did not attenuate mechanical sensitivity when administered after CIPN was induced by paclitaxel treatment. However, KLS-13019 significantly and dose-dependently attenuated tactile sensitivity in the reversal paradigm and was more potent and effective than treatment with morphine. Importantly, KLS-13019 also attenuated the reinforcing properties of morphine in a mouse model of morphine self-administration. In vitro, we have shown that KLS-13019 and CBD protect against paclitaxel-induced oxidative stress in dorsal root ganglia cultures, and that a mechanism underlying this neuroprotection is regulation of intracellular calcium via the mitochondrial Na+/Ca++ exchanger-1 (mNCX-1). Our central hypothesis is that administration of CBD or KLS-13019 preserves Ca2+ homeostasis by promoting activity of the mNCX-1. Furthermore, our new data demonstrates that the putative cannabinoid receptor GPR55 is induced following paclitaxel treatment and contributes to sensory neuron toxicity and inflammation that can be reversed by KLS-13019, but not CBD. These studies support a pro- nociceptive, pro-inflammatory role for GPR55 that mediates pain associated with CIPN. We predict bi-modal pharmacological effects of KLS-13019 that can both increase viability of sensory neurons exposed to paclitaxel acutely and decrease inducible GPR55 that contributes to long-term neuroinflammation. Evidence has been obtained that KLS-13019 is an antagonist to GPR55 as shown in a ß-arrestin assay. In Phase 2, we will optimize the process to prepare KLS-13019, develop analytical methods, optimize formulation, and evaluate in pharmacokinetic studies. A fully battery of genotoxicity, safety pharmacology, toxicokinetic, and toxicology reports will be completed. KLS-13019 will be evaluated in a rat models of CIPN, tolerance, impairment, and abuse liability. At the conclusion of this grant, the data will be submitted to the FDA and a pre-IND meeting will be completed.

Public Health Relevance Statement:
Narrative Chronic pain management continues to be a significant societal need with growing concerns about the efficacy of opiates and the risks of long-term dependence. Mitochondrial dysfunction, calcium dysregulation, oxidative stress, and inflammation have all been implicated in neuropathic pain. Cannabidiol (CBD) is a non-psychoactive component of Cannabis sativa that is effective in both treating neuropathic pain and relieving opiate dependence. However, CBD has severe limitations in terms of potency, safety, and oral bioavailability. Kannalife has solved these problems in its patented series of derivatives that include KLS-13019. This grant will further develop KLS-13019 for advancement to safety pharmacology evaluation, toxicology studies, and human clinical trials.

Project Terms:
Oral Administration; Oral Drug Administration; intraoral drug delivery; Affect; Pain management; Pain Control; Pain Therapy; pain treatment; Anti-Inflammatory Agents; Anti-Inflammatories; Anti-inflammatory; Antiinflammatories; Antiinflammatory Agents; antiinflammatory; Biological Assay; Assay; Bioassay; Biologic Assays; Biological Availability; Bioavailability; Biologic Availability; Physiologic Availability; Calcium; Cannabidiol; High Pressure Liquid Chromatography; HPLC; High Performance Liquid Chromatography; High Speed Liquid Chromatography; Clinical Trials; Canis familiaris; Canine Species; Dogs; Dogs Mammals; canine; domestic dog; Pharmaceutical Preparations; Drugs; Medication; Pharmaceutic Preparations; drug/agent; Spinal Ganglia; Dorsal Root Ganglia; dorsal root ganglion; Grant; Homeostasis; Autoregulation; Physiological Homeostasis; Human; Modern Man; In Vitro; Inflammation; Institutes; Laboratories; Methods; Mitochondria; mitochondrial; Morphine; Infumorph; Kadian; MS Contin; MSir; Morphia; Oramorph; Oramorph SR; Roxanol; Statex SR; nervous system disorder; Nervous System Diseases; Neurologic Disorders; Neurological Disorders; neurological disease; Afferent Neurons; Sensory Cell Afferent Neuron; Sensory Neurons; Pain; Painful; Legal patent; Patents; Patients; Drug Kinetics; Pharmacokinetics; Pharmacology; Problem Solving; Production; Quality of life; QOL; Rattus; Common Rat Strains; Rat; Rats Mammals; Reference Standards; Risk; Role; social role; Safety; Self Administration; Self-Administered; Solvents; Toxicology; Vendor; Work; cannabinoid receptor; cytokine; Mediating; Paclitaxel; Anzatax; Asotax; Bristaxol; Paclitaxel (Taxol); Praxel; Taxol; Taxol A; Taxol Konzentrat; beta-arrestin; arrestin B; ß-arrestin; analytical method; improved; Acute; Chronic; Residual state; Residual; Phase; Medical; Series; Chemicals; Evaluation; Nociception; nociceptive; Opioid; Opiates; Oxidative Stress; analog; Collaborations; Toxicokinetics; Exposure to; Attenuated; Inflammatory; mechanical; Mechanics; Tactile; Dependence; Oral; respiratory; Opiate Dependence; opioid addiction; opioid dependence; opioid dependent; Opiate Addiction; meetings; success; genotoxicity; neuroprotection; neural; relating to nervous system; Toxicities; Toxic effect; Histopathology; Structure; Prevention; Modality; C sativa; C. sativa; Cannabis sativa; Cannabis sativa plant; Reporting; Regulation; Modeling; Property; Cancer Treatment; Malignant Neoplasm Therapy; Malignant Neoplasm Treatment; anti-cancer therapy; anticancer therapy; cancer-directed therapy; cancer therapy; preventing; prevent; Causality; causation; disease causation; Etiology; Dose; Data; in vivo; Antiinflammatory Effect; anti-inflammatory effect; Small Business Technology Transfer Research; STTR; Validation; Preparation; Process; Cardiac; Development; developmental; painful neuropathy; neuropathic pain; neuroinflammation; neuroinflammatory; Impairment; neurotoxicity; neuron toxicity; neuronal toxicity; mitochondrial dysfunction; mouse model; murine model; inflammatory marker; inflammation marker; effective therapy; effective treatment; Biological Markers; bio-markers; biologic marker; biomarker; process optimization; prescription opioid; licit opioid; opiate medication; opioid medication; prescribed opiate; prescribed opioid; prescription opiate; Chemotherapy-induced peripheral neuropathy; Formulation; GPR55 receptor; GPR55; pain sensitivity; preservation; in vivo evaluation; in vivo testing; chronic pain management