3K3A-APC / ZZ Biotech - LARVOL DELTA

Endogenous and therapeutic transport of activated protein c across the blood-retina barrier protects the retina (ASH 2025) - "Our study provides the first in vivo demonstration that APC, but not PC, naturally crosses the BRB intothe retina via a selective, likely receptor-mediated process involving EPCR. These observations establish apreviously unrecognized link between circulating APC and retinal homeostasis and suggest the existenceof an active, receptor-mediated mechanism that allows large protective proteins, like APC, to accessimmune-privileged ocular tissues. It appears that, by leveraging this BRB endogenous transport pathway,the systemically administered APC analog, 3K3A-APC, can cross the BRB and exert potent anti-inflammatory effects in the retina."

Hematological Disorders • Inflammation • Ocular Inflammation • Ophthalmology • Retinal Disorders • Uveitis • ITGAM • PROCR

EPCR-mediated endogenous and therapeutic transport of activated protein C across the blood-retina barrier protects the retina. (PubMed, J Thromb Haemost) - "Our findings establish a fundamental step in investigating the concept of PC as a physiologically relevant, naturally protective protein in the eye. Our results support the feasibility of developing non-invasive, systemic APC-based therapies for retinal diseases."

Journal • Ophthalmology • Retinal Disorders • PROCR

Intravenous 3K3A-Activated Protein C Inhibits Murine Ocular Inflammation and Suppresses Ocular Choroidal Neovascularization (ASH 2023) - "Our study reveals that systemically administered 3K3A-APC effectively inhibits ocular inflammation and reduces CNV formation. Drawing insights from the neuroprotective activities of 3K3A-APC in the brain and CNS and recognizing the retina, particularly the macula, as an extension of the brain, we propose 3K3A-APC as a potential neuroprotective treatment for AMD and other neurodegenerative retinal pathologies. Given the established safety of systemic 3K3A-APC administration in phase 2 clinical trials for ischemic stroke, our findings support further exploring 3K3A-APC as a novel therapeutic approach in ophthalmology."

Preclinical • Age-related Macular Degeneration • Cardiovascular • CNS Disorders • Inflammation • Ischemic stroke • Macular Degeneration • Ocular Inflammation • Ophthalmology • Retinal Disorders • Vascular Neurology • Wet Age-related Macular Degeneration • ITGAM • PROCR

Activated protein C ameliorates diabetes-induced atherosclerosis by sustaining macrophage efferocytosis. (PubMed, Cardiovasc Diabetol) - "In vitro, treatment with activated PC (aPC) or its cytoprotective selective variant (3K3A-aPC) restored high glucose-impaired macrophage efferocytosis...aPC's vasculoprotective effects, including the reduction of plaque size, were abrogated upon MerTK inhibition using morpholinos, underscoring the pivotal role of MerTK in mediating aPC's atheroprotective actions. These findings suggest that impaired TM-PAR1-aPC signaling contributes to defective macrophage efferocytosis in diabetes-associated atherosclerosis and that aPC-based therapies may offer a novel strategy to enhance macrophage function and prevent diabetes induced atherosclerosis."

Journal • Atherosclerosis • Cardiovascular • Diabetes • Inflammation • Metabolic Disorders • APOE • ATF6 • MERTK • PROCR

In vivo neuroprotection in ischemic stroke by activated protein C requires β-arrestin 2. (PubMed, Blood Vessel Thromb Hemost) - "Based on quantitation of brain injuries, 3K3A-APC significantly limited brain injury in control mice to relatively small, localized areas, whereas 3K3A-APC's protection was lost in Arrb2 -/- mice. Thus, the major in vitro mechanism of action that requires β-arrestin 2 for APC/PAR1 biased signaling is central to the in vivo mechanism of action for APC's neuroprotection."

Journal • Preclinical • Cardiovascular • CNS Disorders • Infectious Disease • Ischemic stroke • Septic Shock • Vascular Neurology • ARRB1

RHAPSODY-2: Efficacy and Safety Evaluation of 3K3A-APC in Ischemic Stroke (clinicaltrials.gov) - P3 | N=0 | Withdrawn | Sponsor: ZZ Biotech, LLC | N=1400 ➔ 0 | Not yet recruiting ➔ Withdrawn

Enrollment change • Trial withdrawal • Cardiovascular • Ischemic stroke

Effect of Post Thrombolytic Intracerebral Hemorrhage Volume on 90-day Outcomes in Acute Ischemic Stroke Patients. (PubMed, J Stroke Cerebrovasc Dis) - "We did not observe any independent effect of post thrombolytic ICH volume on death or disability in acute ischemic stroke patients. Although further studies must be done, our data suggest that strategies to prevent ICH expansion such as antifibrinolytic medications and reduction in ICH volume such as surgical evacuation may not reduce death or disability in acute ischemic stroke patients with post thrombolytic ICH."

Journal • Cardiovascular • Cerebral Hemorrhage • Hematological Disorders • Ischemic stroke

Activated Protein C’s Cytoprotection Against Thromboinflammation: A Multi-Omic Study (ISTH 2024) - "Aims: Here, we aimed to elucidate if treatment with 3K3A-aPC, a new variant with enhanced cytoprotection and minimal anticoagulant activity, would mitigate endothelial thromboinflammation and uncover important cytoprotective pathways and biomediators of aPC deranged by injury and shock... Multi-omic results demonstrated responses to injury and shock after exposure for 1 hour. aPC mitigated shock-induced hyperuricemia via de-activation of the polyol pathway by restoring adenosine triphosphate (ATP) and urate levels. It maintained energy homeostasis and energy supply by stabilizing tricarboxylic acid (TCA) cycle, creatine phosphate shuttle, and glycolysis metabolites (Figure 1a)."

Hematological Disorders • Inflammation

Cytoprotective 3K3A-activated protein C and plasma: A comparison of therapeutics for the endotheliopathy of trauma. (PubMed, J Trauma Acute Care Surg) - "Our data shows that FP24, in a post-trauma environment, pre-treatment with 3K3A-aPC can potentially mitigate the EoT to a greater degree than FP24 with or without 3K3A-aPC. Although further exploration is needed, this represents a potentially ideal and perhaps superior therapeutic treatment for the dysregulated thromboinflammation of injured patients."

Journal • Inflammation • Mood Disorders

PRE-RECORDED: BLOOD-BRAIN BARRIER AND VASCULAR SYSTEM AS TREATMENT TARGETS FOR COGNITIVE IMPAIRMENT AND ALZHEIMER'S' DISEASE (ADPD 2024) - "Here, I will briefly review the evidence that 1) BBB dysfunction (e.g., transporters, genetic defects) leads to CNS dysfunction (human monogenic disorders, models); 2) BBB breakdown is an early biomarker of human cognitive dysfunction; 3) APOE4 carriers are distinguished from APOE3 homozygotes by pre-clinical BBB breakdown in the hippocampus and medial temporal lobe which becomes more severe with cognitive impairment, but is not related to Abeta or tau pathology measured in CSF or by PET; 4) Breakdown in the BBB and/or diminished cerebral blood flow (CBF) in regions processing memory/global cognition, attention/executive function and language can predict independently neurodegenerative and white matter tract changes in APOE4 individuals bearing e3/e4 and e4/e4 alleles; 5) BBB and CBF changes can lead to multi-domain cognitive impairments independently of and/or via mediation through brain structural changes; 6) Therapeutic approaches directed at the BBB can slowdown and..."

Alzheimer's Disease • CNS Disorders • Cognitive Disorders • Gene Therapies • MMP9 • SLC2A1

Effect of Post Thrombolytic Intracerebral Hemorrhage Volume on 90-day Outcomes in Acute Ischemic Stroke Patients (ISC 2024) - " We analyzed 110 patents recruited in the Safety Evaluation of 3K3A-APC in Ischemic Stroke (RHAPSODY) trial who received intravenous tissue plasminogen activator (tPA) followed by mechanical thrombectomy (if indicated) and 3K3A-APC or placebo... We did not observe any independent effect of post thrombolytic ICH volume on death or disability in acute ischemic stroke patients. Strategies to prevent ICH expansion such as antifibrinolytic medications and reduction in ICH volume such as surgical evacuation may not reduce death or disability in acute ischemic stroke patients with post thrombolytic ICH."

Clinical • Cardiovascular • Cerebral Hemorrhage • Hematological Disorders • Ischemic stroke

Endothelial Protein C Receptor and 3K3A-Activated Protein C Protect Mice from Allergic Contact Dermatitis in a Contact Hypersensitivity Model. (PubMed, Int J Mol Sci) - "3K3A-aPC reduced CHS severity in wild-type and EPCRKO mice by suppressing immune cell infiltration/activation and inflammatory cytokines. In summary, EPCRKO exacerbated CHS, whereas 3K3A-aPC could reduce the severity of CHS in both EPCRKO and wild-type mice."

Journal • Preclinical • Contact Dermatitis • Dermatitis • Dermatology • Immunology • Inflammation • APC • PROCR

A recombinant signalling-selective activated protein C that lacks anticoagulant activity is efficacious and safe in cutaneous wound preclinical models. (PubMed, Wound Repair Regen) - "A bioengineered APC variant designated 3K3A-APC retains APC's cytoprotective cell signalling actions with <10% anticoagulant activity...However the female pigs exhibited transient and mild local reactions after treatments in week three, which did not impact healing. Overall these preclinical studies support the hypothesis that 3K3A-APC merits future human wound studies."

Journal • Preclinical

Multi-modal detection of changes in MND for the quantitative evaluation of response to 3K3A-APC treatment (ALS-MND 2023) - No abstract available

Multi-omic signature of therapeutic activated protein C analog on APOE4 neurovascular dysregulation (Neuroscience 2023) - "Together these data provide a multi-omic signature of the restorative therapeutic effects of 3K3A-APC. 3K3A-APC may be a promising therapeutic to counteract the effects of APOE4 mediated transcriptomic, proteomic, and functional BBB disruption."

Alzheimer's Disease • CNS Disorders • APOE

Activated protein C analog protects ischemic brain injury in mice via β-arrestin-2-dependent signaling (Neuroscience 2023) - "The effects of 3K3A-APC on neuroprotection and blood-brain barrier protection were lost in β-arrestin-2 null mice. Thus, our data supports that 3K3A-APC reduces ischemic brain injury via β-arrestin-2-dependent signaling pathways."

Preclinical • CNS Disorders • Vascular Neurology • ARRB1

Mitigation of Trauma Induced Endotheliopathy by Activated Protein C: A Potential Therapeutic for Post-Injury Thromboinflammation. (PubMed, J Trauma Acute Care Surg) - "Pre-treatment with 3K3A-aPC, which retains its cytoprotective function but has only ~5% of its anti-coagulant function, abrogates the effects of trauma-induced endotheliopathy. This represents a potential therapeutic treatment for dysregulated thrombo-inflammation for injured patients by minimizing aPC's role in trauma-induced coagulopathy while concurrently amplifying its essential cytoprotective function."

Journal • Hematological Disorders • Inflammation • RAC1 • RHOA

3K3A-Activated Protein C Inhibits Choroidal Neovascularization Growth and Leakage and Reduces NLRP3 Inflammasome, IL-1β, and Inflammatory Cell Accumulation in the Retina. (PubMed, Int J Mol Sci) - "These findings indicate that the anti-inflammatory activities of 3K3A-APC contribute to CNV inhibition. Our study suggests the potential use of 3K3A-APC as a novel multi-target treatment for CNV."

Inflammatory cell • Journal • Age-related Macular Degeneration • Inflammation • Macular Degeneration • Ophthalmology • Retinal Disorders • IL1B • NLRP3

RHAPSODY-2: Efficacy and Safety Evaluation of 3K3A-APC in Ischemic Stroke (clinicaltrials.gov) - P3 | N=1400 | Not yet recruiting | Sponsor: ZZ Biotech, LLC | Initiation date: Apr 2023 ➔ Dec 2023

Combination therapy • Trial initiation date • Cardiovascular • Ischemic stroke

3K3A-Activated Protein C treatment exerts anti-inflammatory and blood barrier stabilizing effects in the retina, and inhibits choroidal neovascularization (ISTH 2023) - "3K3A-APC treatment significantly decreased myeloid and microglia cell number and recruitment to the retinal pigment epithelium (RPE)-choroid area (Figure 1). 3K3A-APC treatment inhibited inflammasome activation, determined by lower levels of NLRP3 and its downstream effector IL-1β. ZO1 clustering in the RPE border, noted in 3K3A-APC treated eyes, suggests a barrier protective effect of 3K3A-APC on outer BRB."

Age-related Macular Degeneration • Cognitive Disorders • Inflammation • Macular Degeneration • Ophthalmology • Retinal Disorders • Wet Age-related Macular Degeneration • IL1B • ITGAM • NLRP3 • TJP1

Analysis of Brain Edema in RHAPSODY. (PubMed, Int J Stroke) - "Existing cerebral edema imaging markers potentially describe two distinct processes, including lesional water concentration (i.e., NWU) and mass effect (MLS, HVR, and CSF volume). These two types of imaging markers may represent distinct aspects of cerebral edema, which could be useful for future trials targeting this process."

Journal • Cardiovascular • CNS Disorders • Ischemic stroke

Analysis of the urinary metabolic profiles in irradiated rats treated with Activated Protein C (APC), a potential mitigator of radiation toxicity. (PubMed, Int J Radiat Biol) - "On the other hand, sub-cohorts of rats that were treated with rat wildtype-APC showed alleviation of radiation toxicities, in part, at the 90-day time point, while rat 3K3A-APC showed partial alleviation of radiation induced metabolic alterations 14 days after irradiation. Taken together, these results show that augmenting the Protein C pathway and activity via administration of recombinant APC may be an effective approach for mitigation of radiation induced normal tissue toxicity."

Journal • Preclinical • Dyslipidemia • Metabolic Disorders • APC

Activated protein C analog protects pericyte-deficient mice from ischemic brain injury (Neuroscience 2022) - "Our data suggest that pericyte deficiency results in greater brain injury, BBB breakdown, and neuronal degeneration in stroked mice and that 3K3A-APC protects the brain from accelerated injury caused by pericyte deficiency. These findings may have implications for treatment of ischemic brain injury in neurological conditions associated with pericyte loss such as those seen during normal aging and in neurodegenerative disorders such as Alzheimer's disease."

Preclinical • Alzheimer's Disease • CNS Disorders • Vascular Neurology • PDGFRB

Parmodulin, an Allosteric Inhibitor of Protease Activated Receptor 1 (PAR1) Protects Sickle Cell Mice from Thromboinflammation and Endothelial Dysfunction (ASH 2022) - "3K3A-APC is a signaling selective variant of APC that activates cytoprotective PAR1 signaling with minimal anticoagulant effects, demonstrated to have a beneficial role in stroke models...In contrast, PM2-mediated allosteric inhibition of thrombin/PAR1 and simultaneous induction of APC-like cytoprotective signaling attenuated levels of TAT, IL-6, HMGB1, and sVCAM in HbSS mice. Future studies towards evaluating the effect of PM2 on downstream targets of PAR1 signaling in sickle mice will provide more insight into the mechanism of this protective effect."

Preclinical • Anemia • Cardiovascular • Genetic Disorders • Hematological Disorders • Immunology • Inflammation • Sickle Cell Disease • HMGB1 • IL6 • PROCR

3K3A-Activated Protein C Prevents Microglia Activation, Inhibits NLRP3 Inflammasome and Limits Ocular Inflammation. (PubMed, Int J Mol Sci) - "Finally, 3K3A-APC treatment inhibited inflammasome activation, determined by lower levels of NLRP3 and its downstream effector IL-1β. Our results highlight the anti-inflammatory properties of 3K3A-APC in ocular inflammation and suggest its potential use as a novel treatment for retinal diseases associated with inflammation."

Journal • CNS Disorders • Immunology • Inflammation • Ocular Inflammation • Ophthalmology • Retinal Disorders • Uveitis • IL1B • NLRP3