Neuropharmacology

The treatment of neurological diseases, especially psychiatric and cognitive disorders, remains an unresolved and unmet therapeutic area on a global scale. Uncovering the mechanisms of these diseases, identifying mediators and potential drug target molecules that play key roles in pathophysiological processes, and conducting preclinical pharmacological studies of drug candidates are of great significance from both basic and applied research perspectives.
Neuroinflammation (central nervous system inflammation, glial cell activation) represents a common mechanism underlying chronic pain—especially neuropathic and migraine pain—and depression, which often occur as comorbidities. Neuroinflammation can trigger various neurodegenerative processes, leading to cognitive impairments, dementia (such as Alzheimer’s and Parkinson’s disease), and retinal damage.
Our work primarily focuses on uncovering the mechanisms of these diseases, identifying mediators and potential drug target molecules that play pivotal roles in pathophysiological processes, and conducting preclinical efficacy studies of drug candidates.

Identification of Fibromyalgia Mechanisms and Determination of New Therapeutic Targets to Enhance Treatment Efficiency

Primary chronic pain is a new concept that describes long-lasting, multifactorial pain conditions causing significant stress, suffering, reduced work capacity, and even loss of working ability. This category includes widespread chronic, diffuse, musculoskeletal spontaneous pain, namely fibromyalgia. Affecting 2-8% of the population, fibromyalgia is the second most common rheumatological disorder. The pain often intensifies in cold conditions and is associated with sleep disturbances, chronic fatigue, depression, short-term memory decline, irritable bowel syndrome, and rheumatoid arthritis. The pathophysiological mechanisms of the disease are still unknown, and effective therapy is not available, making its research a top priority.
Chronic psychosocial distress is an etiological and/or aggravating factor of fibromyalgia, often co-occurring with the most common central nervous system comorbidities. Several common mediators have been identified between stress and pain pathways, with neuroinflammation developing in brain regions involved in pain processing and mood regulation, resulting in central sensitization. This can lead to the persistence and exacerbation of pain. However, the relationships between stress, depression, and pain, as well as the molecular mechanisms and signaling pathways, remain largely unknown.
Given that conventional analgesics are often ineffective and/or cannot be used long-term due to severe side effects, fibromyalgia poses a significant challenge for drug development. The lack of precision-designed clinical studies that consider patient heterogeneity and comorbidities, appropriate methodological approaches, and relevant animal models have hindered breakthroughs in analgesic development in recent decades. We aim to address these challenges through a paradigm shift in pharmacological research.
Our research is based on the recent paradigm shift in pharmacological studies, employing a complex, modern methodological approach that generates hypotheses from human studies and validates them in experimental model systems.
Our primary goal is to identify the peripheral and central sensitization mechanisms responsible for chronic pain and determine their associations with comorbidities and psychological factors. Additionally, we aim to explore mediators, pathways, and networks that play crucial roles in pathophysiological processes and their relationships with common central nervous system comorbidities (chronic anxiety, stress, depression), as well as to identify new analgesic targets.
Our research, involving a broad patient group, employs imaging (fMRI), psychological, and laboratory investigations to map pathophysiological mechanisms, pathways, and networks involved in the development, persistence, and aggravation of pain, as well as interactions between stress and pain. We hope that by uncovering the processes and relationships underlying fibromyalgia, we can identify new drug target molecules, therapeutic opportunities, and directions for drug development and repositioning.

Investigation of Target Molecules and Drug Candidates

Our neuropharmacological studies, which combine the high-level research infrastructure of the Szentágothai János Research Center with a strong clinical background, focus on exploring several so-called target molecules, i.e., potential drug targets. These molecules may play a significant role in identifying and treating various neurodegenerative diseases (e.g., Alzheimer's disease, Parkinson's disease), nervous system damage, neurogenic pain, and inflammatory processes.
Some of our studies are already in the drug development phase with the involvement of industrial partners, and our foundational research methods and measurement procedures, based on our basic research results, play an important role in the drug development programs of our key partner, Richter Gedeon Plc.

Research and Development Results Achieved Since the Start of the Laboratory

Cognitive Behavioral Tests on Primates:
We developed a new free-choice sequential visual test task (SOSS) to measure working memory and conducted pharmacological validation using a performance-impairing agent (scopolamine), making the SOSS task available for further studies in our laboratory.
We conducted an efficacy study of a new bioactive compound (patented by an industrial partner) in an object-location working memory task (vsPAL).
We designed and established a new translational cognitive test (human-monkey) by organizing four cognitive tasks (SOSS, DMTS, vsPAL, IDED) into a joint test package for rhesus macaque monkeys.

Quantitative Pharmacological EEG Studies on Primates:
We developed a non-invasive pharmacological EEG methodology to study the pharmacodynamic effects of drug candidates on the brain and established quantitative brain oscillation measurement techniques in a human-like measurement system (scalp EEG) on rhesus macaque monkeys.
We validated the test system using known neuromodulatory agents, confirming its suitability for further research tasks.

Functional Cortical Stimulation Studies on Primates:
We established a preclinical functional neuromodulation environment (transcranial magnetic stimulation, TMS) to determine and transiently influence cortical excitability in rhesus macaque monkeys.
In pharmacological studies with known neuromodulators, we successfully demonstrated the dose-dependent change in motor threshold.
We optimized TMS technology for examining cortical excitability (I/O curve recording, short-interval cortical inhibition paradigm, SICI) and for inducing virtual lesions.

Establishment of a National Research Network:
We have initiated the organization of a national research network centered around the Endre Grastyán Translational Research Center, involving academic partners in addition to our main industrial partner, Richter Gedeon Plc.

Significance of the Work Performed:
We have gained new insights into the fundamental mechanisms of neurocognitive and psychiatric diseases. Additionally, the established research methodology has been made suitable for identifying mediators and potential drug target molecules that play key roles in cognitive processes, as well as for conducting preclinical efficacy studies of drug candidates.
In the long term, the results may contribute to a deeper understanding of cortical functions in primates, modeling higher-order (psychiatric) nervous system diseases, and discovering new therapeutic opportunities, potentially offering significant domestic economic and market advantages.

Dissemination of Results:
The results have been presented in numerous conference talks and posters, and three scientific publications are in preparation, one of which is under revision in a D1 journal (Neuroimage).
The methodological developments and investigations are also part of the research topics of two PhD students.

2.2. Research and Development (R&D) Results Achieved During the Reporting Period

I. In Vivo Behavioral Pharmacology Method Development and Drug Efficacy Studies:
We continued the development of translational joint (human-non-human) primate complex (electrophysiological, neuromodulation, and behavioral) test paradigms, focusing on the following areas:

1. Cognitive Behavioral Testing on Primates:
   During this phase, we continued the development of three previously validated primate-specific cognitive tasks (SOSS, DMTS, vsPAL) to study the cognitive effects of a small-molecule compound developed by our industrial partner.
   We further developed the "humanized" cognitive behavioral test package paradigm. Initially, the animals were trained to switch between test tasks on different days, and later within the same day.
   We demonstrated that random task switching did not affect the animals’ performance, allowing us to create experimental conditions highly similar to those used in human clinical trials (achieving high translational validity).
2. Examination of Cortical Excitability and Virtual Lesion Induction using Transcranial Magnetic Stimulation (TMS) on Primates:
   We continued setting up the paired-pulse inhibition (PPI) protocol and repetitive TMS (rTMS) protocols to externally, non-pharmacologically, and reversibly impair cognitive performance (virtual lesion technique).
   Pilot studies with a small sample size were conducted to assess the protocols.
   We successfully established the PPI protocol, and the optimization of the repetitive stimulation sequence (cTBS) is ongoing.
   During this phase, we received a new TMS device equipped with a coil capable of delivering sham (placebo) pulses and integrated software, enabling double-blind clinical trial protocols.
   Training by the supplier was completed, and internal testing and setup of the device have begun.

II. Organization of the National Research Network:
In accordance with our plans, we have initiated the establishment of a research network centered around the Endre Grastyán Translational Research Center.
During the reporting period, we also prepared a major industrial consortium research proposal, which Richter Gedeon Plc. will submit.

2.3. Expected R&D Results in the Next Reporting Period

I. In Vivo Behavioral Pharmacology Method Development and Drug Efficacy Studies:
We will continue methodological developments and efficacy studies of selected drug candidate compounds that are on the verge of the clinical trial phase, using macaque monkeys.

1. Pharmacokinetic and Cognitive Behavioral Studies on Primates:
   Following previous phases, we plan to utilize the previously validated high-throughput primate-specific cognitive tasks to study small-molecule compounds developed by our industrial partner.
   Over the next six months, we plan to investigate three newly patented compounds.
   During the next phase, we will continue the development of the "humanized" cognitive behavioral test package, initially with random switching between two, then four tasks within a single session.
   If successful, we will begin the pharmacological validation of the paradigm using test compounds known to influence cognitive performance (e.g., transient amnesic agent - scopolamine).
2. Examination of Cortical Excitability and Virtual Lesion Induction using TMS on Primates:
   During this phase, we plan to set up the new TMS stimulation device capable of double-blind testing and begin the pharmacological validation of a new paired-pulse inhibition (PPI) protocol (short-interval cortical inhibition, SICI).
   Additionally, we will continue setting up the rTMS protocol to externally, non-pharmacologically, and reversibly impair cognitive performance in primates (virtual lesion technique).
   Upon successful setup, we will validate the system using the established test package from point 1.

II. Expansion of the National Research Network:
In the next phase, we will continue to expand the research network around the Endre Grastyán Translational Research Center and develop joint research projects with the GTKK and participating partners, aligned with the professional tasks undertaken in TINL.