The 20S proteasome constitutes one of the vital cellular proteolytic machineries that maintain protein homeostasis (proteostasis) and participates in almost all cellular functions through the degradation of misfolded, redundant and damaged proteins.  The 20S proteasome consists of two α-rings which control the access to the proteolytic sites located in the two inner β-rings.
Proteasome has been reported to decline in terms of quantity and function during ageing and age-related diseases progression. Thus, proteasome activation constitutes a central mechanism for the deceleration of organismal aging and of the progression of neurodegenerative diseases such as Alzheimer's and Huntington's diseases, which are related with accumulation of toxic protein aggregates.

Although the research towards the development of small molecules as proteasome activators is scarce, the results so far suggest that activation of the proteasome complex constitutes a pioneer strategy for the deceleration of ageing and age-related diseases.
A novel class of hybrid compounds that combine structural features of the natural antioxidant vitamin E and of hydroxytyrosol connected through carefully selected linkers, was designed and synthesized by our group, leading to the development of an optimum structural proteasome activator (MK151) which improved healthspan markers in cellulo and in C. elegans model.
Based on these findings 16 novel hybrid compounds were designed and synthesized. Among them, 5 chroman analogues and 2 benzoxathiine (bioisostere of chroman ring) analogues were identified as structural activators of purified 20S proteasome complex. These compounds represent a new class of structural proteasome activators.

This class of compounds was further exploited by synthesizing more analogues. The biological results to date, from Dr. N. Chondrogianni’s group, show great promise with regards to the anti-ageing and neuroprotective effects of the new analogues in cellulo and in organismal level. Moreover, the most studied structural proteasome activator MK151 was synthesized on kg scale and incorporated in cosmetic preparations.

Supported by the following research grants: STHENOS-b 2017-2020 collaboration with members of other research teams of our Institute

RESET: “Bio-inspired antiaging proteasome activators” In collaboration with Intermed SA.

Inflammation is an important factor in promoting various diseases.  An increase of the proinflammatory cytokine interleukin-1β (IL-1β) levels has been associated to the pathogenesis inflammatory diseases such as cardiovascular disease (CVD) and cancer.
NLRP3 inflammasome, a multi-protein complex triggering the maturation of the proinflammatory cytokine IL-1β, has attracted tremendous research interest. NLRP3 assembly and IL-1β expression are promoted by extracellular ATP binding as a danger signal to purinergic P2X7 receptor. Currently no therapeutic agent able to hit different steps of inflammasome activation is available on the market. Thus, more than 30 novel compounds targeting P2X7R, were designed and synthesized in the context of D. Pournara’s PhD thesis and tested by Prof. Annette Nicke, Ludwig-Maximilians University in Munich, Germany
Supported by: Alexander S. Onassis Public Benefit Foundation fellowship.

NLRP3 activation has been linked to the pathophysiology of CVD. One of the major underlying factors for CVD is atherosclerosis, a multi-step chronic disease, characterized by a close interrelation between inflammation and thrombotic lesions that may lead to myocardial infarction. However, there is limited knowledge about targeting IL-1β against atherosclerosis, while current antithrombotic therapies are associated with a high bleeding risk.
In collaboration with Assoc. Prof. Karin H Franzén and Prof. Magnus Grenegård from Cardiovascular Research Centre (CVRC) at Örebro University (ÖU) Sweden, we revealed that two 6-piperazinyl-purine analogues synthesized by our group, inhibit the release of proinflammatory cytokines like IL-1 β and the expression of NLRP3 inflammasome genes and proteins, imparting an anti-inflammatory effect in aortic smooth muscle cells (AoSMCs).
Moreover, the involvement of kinases such as JAK and ROCK, in the pathogenesis of CVD could provide an exciting avenue for future research. Recent results of our fruitful collaboration with the CVRC, showed thatRho-associated protein kinase is the main molecular target for the anti-thrombotic activity of one 6-piperazinyl-purine analogue synthesized by our group.
Supported by: Swedish Knowledge foundation HÖG19 (KKS-HÖG19)

In the context of a recently approved Knowledge Foundation grant, Örebro University, seven Swedish biotech companies and three collaborating institutes will investigate the effects of new drug candidates against CVD and breast cancer. The novel drug candidates will be evaluated in cell and tissue models at Örebro University and the ICB/NHRF is a collaborating institute in the project and will contribute to the project with our expertise in chemical synthesis and in silico analysis, to learn more about the mechanisms of action of these drug candidates.
Supported by: Swedish Knowledge foundation Synergi21 (KKS-Synergi21)

1988: PhD, University of Geneva, Faculty of Sciences, Switzerland.
Title: "Désoxyhydroxyaminosucres par cyclisation ou réduction des nitrones et radicaux libres correspondants. Nouvelles voies de synthèse d’oligosaccharides"

• Pournara D.T., Durner A., Kritsi E., Papakostas A., Zoumpoulakis P., Nicke A., Koufaki M., Design, Synthesis, and in vitro Evaluation of P2X7 Antagonists. ChemMedChem 2020, 15, 2530–2543  
  DOI: https://doi.org/10.1002/cmdc.202000303
• Kardeby, C., Paramel, G.V., Pournara, D., Fotopoulou, T., Sirsjö, A., Koufaki, M., Fransén, K., Grenegård, M., A novel purine analogue bearing nitrate ester prevents platelet activation by ROCK activity inhibition, Eur. J. Pharmacol. 2019, 857, 172428   
  DOI: https://doi.org/10.1016/j.ejphar.2019.172428
• Smiljkovic M., Matsoukas M.-T., Kritsi E., Zelenko U., Golic Grdodolnik S., Calhelha R., Ferreira I., Sankovic-Babic S., Glamoclija J., Fotopoulou T., Koufaki M., Zoumpoulakis P., Sokovic M. Nitrate esters of heteroaromatic compounds as novel Candida albicans CYP51 enzyme inhibitors. ChemMedChem 2018, 13, 251-258
  DOI: https://doi.org/10.1002/cmdc.201700602
• Pournara D., Heropoulos G.A. Koufaki M.* Convenient method for the synthesis of 5-(4-methoxyphenyl)-3H-1,2-dithiole-3-thione (ADT-OMe) and 5-(4-hydroxy phenyl)-3H-1,2-dithiol-3-thione (ADT-OH) using microwave irradiation. Tetrahedron Lett. 2017, 58, 2378-2380     
  DOI: http://dx.doi.org/10.1016%2Fj.tetlet.2017.05.010
• Fotopoulou T., Ćirić A., Kritsi E., Calhelha R.C., Ferreira I.C.F.R., Soković M., Zoumpoulakis P., Koufaki M.  Antimicrobial/Antibiofilm Activity and Cytotoxic Studies of β-Thujaplicin Derivatives. Archiv. Pharm. 2016, 698-709  
  DOI: https://doi.org/10.1002/ardp.201600095
• Koufaki M.*, Fotopoulou T., Kapetanou M., Heropoulos G.A., Gonos E.S., Chondrogianni N. Microwave-assisted synthesis of 3,5-disubstituted isoxazoles and evaluation of their anti-ageing activity. Eur. J. Med. Chem.2014, 83, 508-515    
  DOI: https://doi.org/10.1016/j.ejmech.2014.06.046
• Maugé L., Fotopoulou T., Delemasure S., Dutartre P., Koufaki M., Connat J.-L. In vitro inflammatory/anti-inflammatory effects of nitrate esters of purines. Eur. J. Pharmacol. 2014, 730, 148-156    
  DOI: https://doi.org/10.1016/j.ejphar.2014.02.022
• Koufaki M.*, Fotopoulou T., Heropoulos G.A. Synergistic effect of dual-frequency ultrasound irradiation in the one-pot synthesis of 3,5-disubstituted isoxazoles. Ultrason. Sonochem 2014, 21, 35-39     
  DOI: https://doi.org/10.1016/j.ultsonch.2013.05.012
• Koufaki M.*, Fotopoulou T., Iliodromitis E. K., Bibli S. I., Zoga A., Kremastinos D.Th., Andeadou I.  Discovery of 6-[4-(6-nitroxyhexanoyl)piperazin-1-yl)]-9H-purine, as pharmacological post-conditioning agent. Bioorg. Med. Chem. 2012, 20, 5948-5956  
  DOI: https://doi.org/10.1016/j.bmc.2012.07.037
• Koufaki M.*, Tsatsaroni A., Alexi X., Guerrand H., Zerva S., Alexis M.N. Isoxazole substituted chromans against oxidative stress-induced neuronal damage. Bioorg. Med. Chem. 2011, 19, 4841-4850    
  DOI: https://doi.org/10.1016/j.bmc.2011.06.074
• Koufaki M.*, Theodorou E., Alexi X., Alexis M.N. Synthesis of a second generation chroman/catechol hybrids and evaluation of their activity in protecting neuronal cells from oxidative stress-induced cell death. Bioorg. Med. Chem. 2010, 18, 3898-3909    
  DOI: https://doi.org/10.1016/j.bmc.2010.04.042
• Koufaki M.*, Theodorou E., Alexi X., Nikoloudaki F., Alexis M.N. Synthesis of tropolone derivatives and evaluation of their in vitro neuroprotective activity. Eur. J. Med. Chem.2010, 45, 1107-1112    
  DOI: https://doi.org/10.1016/j.ejmech.2009.12.006
• Koufaki M.*, Kiziridi C., Alexi X., Alexis M.N. Design and synthesis of novel neuroprotective 1,2-dithiolane/chroman hybrids.  Bioorg. Med. Chem., 2009, 17, 6432-6441  
  DOI: https://doi.org/10.1016/j.bmc.2009.07.010
• Melagraki G., Afantitis A., Igglessi-Markopoulou O., Detsi A., Koufaki M., Kontogiorgis C., Hadjipavlou-Litina D.J. Synthesis and evaluation of the antioxidant and anti-inflammatory activity of novel coumarin-3-aminoamides and their alpha-lipoic acid adducts. Eur. J. Med. Chem. 2009, 44, 3020-3026     
  DOI: https://doi.org/10.1016/j.ejmech.2008.12.027
• Koini E.N., Papazafiri P., Vassilopoulos A., Koufaki M., Horváth Z., Koncz I., Virág L., Papp G.J., Varró A., Calogeropoulou T. 5,7,8-trimethyl-benzopyran and 5,7,8-trimethyl-1,4-benzoxazine aminoamide derivatives as novel antiarrhythmics against ischemia-reperfusion injury. J. Med. Chem. 2009, 52, 2328-2340     
  DOI: https://doi.org/10.1021/jm801228h
• Koufaki M.*, Detsi A.; Kiziridi C. Multifunctional lipoic acid conjugates. Curr. Med. Chem. 2009, 16, 4728-4742  
  DOI: https://doi.org/10.2174/092986709789878274
• Andreadou I., Iliodromitis E.K., Farmakis D., Koufaki M., Tsotinis A., Kremastinos D.Th. Alternative pharmacological interventions that limit myocardial infarction Curr. Med. Chem. 2008, 15, 3204-3213  
  DOI: https://doi.org/10.2174/092986708786848550
• Andreadou I., Koufaki M., Iliodromitis E., Kremastinos D.Th. Pharmacological pre- and post-conditioning agents: reperfusion-injury of the heart revisited. Mini Rev. Med.Chem. 2008, 8, 952-959   
  DOI: http://dx.doi.org/10.2174/138955708785132819
• Fotopoulou T., Iliodromitis E. K., Koufaki M.*, Tsotinis A., Zoga A., Gizas V., Pyriochou A., Papapetropoulos A., Andreadou I., Kremastinos D.Th. Design and synthesis of nitrate esters of aromatic heterocyclic compounds as pharmacological preconditioning agents. Bioorg. Med. Chem. 2008, 16, 4523-4531    
  DOI: https://doi.org/10.1016/j.bmc.2008.02.051
• Koufaki M.*, Kiziridi C., Nikoloudaki F., Alexis M.N. Design and synthesis of 1,2-dithiolane derivatives and evaluation of their neuroprotective activity. Bioorg. Med. Chem. Lett. 2007, 17, 4223-4227    
  DOI: https://doi.org/10.1016/j.bmcl.2007.05.036
• Kapou A., Avlonitis N., Detsi A., Koufaki M., Calogeropoulou T., Benetis N.P., Nikolaropoulos S., Mavromoustakos T. CoMFA and CoMSIA 3D Quantitative Structure-Activity Relationships of New Flexible Antileishmanial Ether Phospholipids. Bioorg. Med. Chem. 2007, 15, 1252-1265   
  DOI: https://doi.org/10.1016/j.bmc.2006.11.018
• Detsi A, Bouloumbasi D., Prousis K.C., Koufaki M., Athanasellis G., Melagraki G., Afantitis A., Igglessi-Markopoulou O., Kontogiorgis C., Hadjipavlou-Litina D.J. Design and synthesis of novel quinolinone-3-aminoamides and their α-lipoic acid adducts as antioxidant and anti-inflammatory agents. J. Med. Chem. 2007, 50, 2450-2458    
  DOI: https://doi.org/10.1021/jm061173n
• Koufaki M.*, Kiziridi C., Papazafiri P., Vassilopoulos A., Varró A., Nagy Z., Farkas A., Makriyannis A. Synthesis and biological evaluation of benzopyran analogues bearing class III antiarrhythmic pharmacophores. Bioorg. Med. Chem. 2006, 14, 6666-6678     
  DOI: https://doi.org/10.1016/j.bmc.2006.05.065
• Koufaki M.*, Theodorou E., Galaris D., Nousis L., Katsanou E. S, Alexis M.N. Chroman/catechol hybrids: Synthesis and evaluation of their activity against oxidative stress induced cellular damage. J. Med. Chem. 2006, 49, 300-306   
  DOI: https://doi.org/10.1021/jm0506120
• Papazafiri P., Avlonitis N., Angelou A., Calogeropoulou T., Koufaki M., Scoulica E., Fragiadaki I. Structure-activity relationships of antineoplastic ring-substituted ether phospholipid derivatives. Cancer Chemother. Pharmacol. 2005, 56, 261-270      
  DOI: https://doi.org/10.1007/s00280-004-0935-6
• Koufaki M.*, Detsi A., Theodorou E., Kiziridi C., Calogeropoulou T., Vassilopoulos A., Kourounakis A., Rekka E., Kourounakis P., Gaitanaki C., Papazafiri P. Synthesis of Chroman Analogues of Lipoic Acid and Evaluation of their Activity against Reperfusion Arrhythmias. Bioorg. Med. Chem. 2004, 12, 4835-4841 
  DOI: https://doi.org/10.1016/j.bmc.2004.07.012
• Calogeropoulou Τ., Detsi Α., Lekkas Ε, Koufaki Μ. Strategies in the design of prodrugs of anti HIV agents. Curr. Top. Med. Chem., 2003, 3, 1467-1495  
  DOI: http://dx.doi.org/10.2174/1568026033451763 
  Guest Editor Maria Koufaki.Title of the issue: "Therapeutic strategies against HIV infection"
• Koufaki M.*, Calogeropoulou T., Rekka E., Chryselis M., Papazafiri P., Gaitanaki C., Makriyannis A. Bifunctional Agents for Reperfusion Arrhythmias.  Novel Hybrid Vitamin E/Class I Antiarrhythmics. Bioorg. Med. Chem. 2003, 11, 5209-5219 
  DOI: https://doi.org/10.1016/j.bmc.2003.08.010
• Avlonitis N., Lekka E., Detsi A., Koufaki M., Calogeropoulou T., Skoulica E., Siapi E., Kyrikou I., Mavromoustakos T., Tsotinis A., Golic Grdadolnik S., Makriyannis A.  Antileishmanial Ring-Substituted Ether Phospholipids. J. Med. Chem. 2003, 46, 755-767
  DOI: https://doi.org/10.1021/jm020972c
• Detsi A.,Koufaki M., Calogeropoulou T. Synthesis of (Z)-4-hydroxytamoxifen and (Z)-2-(4-[1-(p-hydroxyphenyl)-2-phenyl]-1-butenyl)phenoxyacetic acid. J. Org. Chem. 2002, 67, 4608-4611   
  DOI: https://doi.org/10.1021/jo0255328
• Koufaki M.*, Calogeropoulou T., Detsi A., Roditis A., Kourounakis A., Papazafiri P., Tsiakitzis K., Gaitanaki C., Beis I., Kourounakis P. Novel potent inhibitors of lipid peroxidation with protective effects against reperfusion arrhythmias. J. Med. Chem. 2001, 44, 4300-4303   
  DOI: https://doi.org/10.1021/jm010962w
• Mavromoustakos T., Calogeropoulou T., Koufaki M., Kolocouris A., Demetzos K., Meng Z., Makriyannis A., Balzarini J., De Clercq E. Ether phospholipid-AZT conjugates possessing anti-HIV and anti-tumor cell activity. Synthesis, conformational analysis and study of their thermal effects on membrane bilayers.  J. Med. Chem.  2001, 44, 1702-1709    
  DOI: https://doi.org/10.1021/jm001121c
• Tronchet J.M.J., Koufaki M., Barbalat F., Geoffroy M. Blocked Sugar Analogues Bearing two or three N(sp3)-O Bonds. Carbohydr. Lett. 1999, 3, 255-262
• Mavromoustakos T., Theodoropoulou E., Yang D.P., Lin S.Y., Koufaki M., Makriyannis A.  The conformational properties of the antineoplastic ether lipid 1- thiohexyl-2-O-methyl-S-glycero-3-phosphocholine. Chem. Phys. Lipids 1996, 84, 21-34    
  DOI: https://doi.org/10.1016/S0009-3084(96)02615-1
• Koufaki M., Polychroniou V., Calogeropoulou T., Tsotinis A., Drees M., Fiebig H.H., LeClerc S., Hendriks H.R., Makriyannis A. Alkyl and alkoxyethyl antineoplastic phospholipids. J. Med. Chem. 1996, 39, 2609-2614 
  DOI: https://doi.org/10.1021/jm9509152
• Koufaki M.*, Calogeropoulou T., Mavromoustakos T., Theodoropoulou E., Tsotinis. A., Makriyannis A. Synthesis of 2H-Labelled Alkoxyethyl Phosphodiester AZT Derivatives for Solid-State 2H-NMR Studies. J. Het. Chem. 1996, 33, 619-622  
  DOI: https://doi.org/10.1002/jhet.5570330316
• Tsotinis A., Calogeropoulou T., Koufaki M., Souli Ch., Balzarini J., De Clercq E., Makriyannis A. Synthesis and Antiretroviral Evaluation of New Alkoxy and Aryloxy Phosphate Derivatives of AZT. J. Med. Chem. 1996, 39, 3418-3422
  DOI: https://doi.org/10.1021/jm950777g
• Calogeropoulou T., Koufaki M., Tsotinis A., Balzarini J., De Clercq E., Makriyannis A. Synthesis and anti-HIV evaluation of alkyl and alkoxyethyl phosphodiester AZT derivatives. Antiv. Chem. Chemother. 1995, 6, 43-49
• Tronchet J.M.J., Koufaki M., Zosimo-Ladolfo G., Bernardinelli G. Spiro naphthoxazine derivatives of furanose sugars. J. Chem. Research 1992 (M) 2501, J. Chem. Research1992 (S) 293
• Tronchet J.M.J., Koufaki M., Zosimo- Ladolfo G. Spacer arms bearing two N- hydroxyimino groups and nitroxyl free radicals thereof. Carbohydr. Res. 1991, 209,   299 
  DOI: https://doi.org/10.1016/0008-6215(91)80167-L
• Tronchet J.M.J., Bizzozero N., Koufaki M., Habashi F., Geoffroy M. Analogs of blocked disaccharides bearing a N-hydroxyimino bridge and nitroxyl free radicals thereof. J. Chem. Research  1989 (M) 2601 J. Chem. Research 1989 (S)  334

• Koufaki M.  Vitamin E derivatives: a patent review (2010-2015). Expert Opin. Ther. Pat. 2016, 26, 35-47
  DOI: https://doi.org/10.1517/13543776.2016.1106476
• Koufaki M. Therapeutic applications of lipoic acid: a patent review (2011-2014) Expert Opin. Ther. Pat. 2014, 24, 993-1005  
  DOI: https://doi.org/10.1517/13543776.2014.937425

• Vasilopoulou M.A., Fotopoulou T., Papaevgeniou N., Gioran A., Papahatjis D., Calogeropoulou T., Koufaki M., Chondrogianni N. A novel bioinspired proteasome activator against aging Free Radic. Biol. Med. 2021, 177, S122-S123
• Papaevgeniou N., Fotopoulou T., Vasilopoulou M., Panat N., Pick E., Golan A., Gonos E.S., Papahatjis D., Calogeropoulou T., Koufaki M., Chondrogianni N. A novel bioinspired proteasome activator: potential anti-ageing strategies offered by mother nature (but not only). Free Radic Biol Med. 2019, 139, S78-S79
• Andeadou I., Bibli S.I., Zhou Z., Fotopoulou T., Koufaki M., Anastasiou-Nana M.,  Papapetropoulos A., Iliodromitis E.K. Pharmacological characterization and elucidation of the molecular mechanism of action of a novel purine analogue which has superior effects to ischemic postconditioning. Eur. Heart J. 2014, 35S1, 386-387 
• Andeadou I., Fotopoulou T., Georgakopoulos A., Zoga A., Koufaki M., Kremastinos D.Th., Iliodromitis E.K., Anastasiou-Nana M. Nitric oxide donors as pharmacological postconditioning agents: the formation of peroxynitrite during reperfusion plays a critical role. Eur. Heart J. 2011, 32S1, 331-332
• Andreadou I., Fotopoulou T., Koufaki M., Tsotinis A., Zoga A., Pyriochou A., Farmakis D., Iliodromitis E.K., Kremastinos D.Th. Novel pharmacological compounds as triggers of preconditioning in vivo. J. Am. Coll. Cardiol. 2008, 51, Suppl. A, A183
• Andreadou I., Fotopoulou T., Koufaki M., Tsotinis A., Zoga A., Papapetropoulos A., Iliodromitis E.K., Kremastinos, D.Th.  Development of novel pharmacological agents that precondition the ischemic heart in vivo through parallel and combined molecular mechanisms Eur. Heart J. 2008, 29, Suppl. 1, 110-111
• Andreadou I., Fotopoulou T., Koufaki M., Tsotinis A., Zoga A., Gizas V., Iliodromitis E., Kremastinos D.Th. Novel pharmacological compounds as triggers of ischemic preconditioning in vivo. J. Mol. Cell. Cardiol. 2007, 42, S1, S178

Maria Koufaki and Anastasia Detsi.  Design and Synthesis of Antioxidant α-Lipoic Acid Hybrids. Advanced Protocols for Oxidative Stress II Series: Methods in Molecular Biology, Vol. 594, Armstrong, Donald (Ed.) 2010, p. 297-309 DOI 10.1007/978-1-60761-411-1_21

• GR 20180100094, WO2019171088A1 Applicant: NHRF, EP3761950A1, Applicant: Ioulia and Irene Tseti Pharmaceutical Laboratories SA. Inventors: Koufaki M., Calogeropoulou T., Chondrogianni N., Papahatjis D., Gonos E.S., Fotopoulou T., Proussis K., Chazapi E. “Bio-inspired proteasome activators with anti-ageing activity”
• GR20050100100. Calogeropoulou T., Koufaki M. “Ultrasound contrast agents containing anticancer ether phospholipids”
• US2003/34225, WO2004041167, CN1742017, CA2503058, AU2003285071, EP1578754, US2006105998 Calogeropoulou T., Koufaki M., Avlonitis N., Makriyannis A."Antiprotozoal ring-substituted phospholipids"
• WO0204438, US2004259763, CA2415523, EP1301504, DE60106922T, ΕS231514Τ Applicant: UNI-PHARMA SA Inventors: Koufaki M., Calogeropoulou T., Makriyannis A. "Bifunctional agents possessing antioxidant and antiarrhythmic activity"