department of neurology, school of medicine, university of

Postępowanie habilitacyjne: Dr n. med. Dorota Żółkowska Załącznik 3

1

Autoreferat

Dr n. med. DOROTA ŻÓŁKOWSKA

Department of Neurology, School of Medicine,

University of California, Davis, Sacramento, CA USA


2

1. Name and Surname: Dorota Zolkowska

2. Diplomas, degrees in science/ art – with the name, place and the year and doctoral

thesis title

1999 1st degree of board certification in Internal Medicine, excellent result

1999 Ph.D. in Pharmacology,

Medical University, Lublin, Poland

Doctoral thesis title: “The effects of diazepam and clonazepam on rat’s

platelet and neutrophil function”

1994 M.D., with honors, Medical University, Lublin, Poland

3. Previous employment in the scientific/ art institutions:

2007-present Project Scientist, Department of Neurology, School of Medicine, University of

California, Davis, Sacramento, CA, USA) and part of the UC Davis

CounterACT Center of Excellence.

http://www.ucdmc.ucdavis.edu/neurology/faculty/

http://counteract.ucdavis.edu/project-core/project-1-prevention-termination-

seizures

2012-present Guest Researcher, Medicinal Chemistry Section, Molecular Targets and

Medications Discovery Branch, National Institute on Drug Abuse, Baltimore,

MD, USA.

2010-2012 Guest Researcher, Clinical Psychopharmacology Section, Medications

Discovery Branch, National Institute on Drug Abuse, Baltimore, MD, USA.

2004-2007 Visiting Fellow, Clinical Psychopharmacology Section, Medications Discovery

Branch, National Institute on Drug Abuse, Baltimore, MD, USA.

2002-2004 Visiting Fellow, Integrative Neuroscience Section, Behavioral Neuroscience

Branch, National Institute on Drug Abuse, Baltimore, MD, USA.

2001-2008 Assistant Professor, Department of Hygiene, Medical University, Lublin,

Poland.

1999-2001 Research Associate, Department of Hygiene, Medical University, Lublin,

Poland.

1995-1999 Doctoral Fellow in Ph.D. Program, Department of Pharmacology and

Toxicology, Medical University, Lublin, Poland.

1994-1995 Postgraduate Internship, Provincial Hospital (Wojewodzki Szpital Zespolony

im. Jana Bozego), Lublin, Poland.

http://www.ucdmc.ucdavis.edu/neurology/faculty/

http://counteract.ucdavis.edu/project-core/project-1-prevention-termination-seizures



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4. Achievement indication under Art. 16 paragraph 2 of the act from March 14th 2003

on academic degrees and academic titles in science and on degrees and titles in art.

(Dz. U. No 65, item 595 as amended):

a) Title of the scientific/artistic achievement

The effects of compounds that affect central nervous system neurotransmission

on the activity of antiepileptic drugs in experimental model of tonic-clonic

seizures.

b) Publications that are a part of the scientific achievement (author/authors, title/titles

of publication, year of publication, name of publisher)

1. Zolkowska D, Kondrat-Wrobel MW, Florek-Luszczki M, Luszczki JJ. Influence of

MPEP (a selective mGluR5 antagonist) on the anticonvulsant action of novel

antiepileptic drugs against maximal electroshock-induced seizures in mice. Prog

Neuropsychopharmacol Biol Psychiatry. 2016;65:172-178.

IF=3,689; MNiSW=35 pkt. (original article)

My input was related to the overall concept of the project, the analysis and

interpretation of the results, the statistical analysis, as well as the literature review

and manuscript preparation. My estimated contribution to the publication was 80%.

2. Zolkowska D, Andres-Mach M, Prisinzano TE, Baumann MH, Luszczki JJ. Modafinil

and its metabolites enhance the anticonvulsant action of classical antiepileptic drugs

in the mouse maximal electroshock-induced seizure model. Psychopharmacology

(Berl). 2015;232(14):2463-2479.





3. Zolkowska D, Kominek M, Florek-Luszczki M, Kocharov SL, Luszczki JJ. Effects of

N-(morpholinomethyl)-p-isopropoxyphenylsuccinimide on the protective action of

different classical antiepileptic drugs against maximal electroshock-induced tonic

seizures in mice. Pharmacol Rep. 2013;65:389-398.





4. Luszczki JJ, Kominek M, Florek-Luszczki M, Tchaytchian DA, Kocharov SL,

Zolkowska D. Influence of N-hydroxymethyl-p-isopropoxyphenylsuccinimide on the

anticonvulsant action of different classical antiepileptic drugs in the mouse maximal

electroshock-induced seizure model. Epilepsy Res. 2012;100:27-36.



4




5. Luszczki JJ, Jaskolska A, Dworzanski W, Zolkowska D. 7-nitroindazole, but not NG-

nitro-L-arginine, enhances the anticonvulsant activity of pregabalin in the mouse

maximal electroshock-induced seizure model. Pharmacol Rep. 2011;63:169-175.





6. Luszczki JJ, Cioczek JD, Kocharov SL, Andres-Mach M, Kominek M, Zolkowska D.

Effects of three N-(carboxyanilinomethyl) derivatives of p-

isopropoxyphenylsuccinimide on the anticonvulsant action of carbamazepine,

phenobarbital, phenytoin and valproate in the mouse maximal electroshock-induced

seizure model. Eur J Pharmacol. 2010;648:74-79.





Summarized Impact Factor of 6 publications 17,152

MNiSW points 167 pkt

c) Description of scientific/artistic aims of work mentioned above and obtained results

with the discussion of their possible application.

The term “epilepsy” encompasses many etiologically different diseases that are

characterized by epileptic seizures. Although the cause of most cases of epilepsy is

unknown, genetic predisposition, brain injury, strokes, brain tumors, infections of the brain,

alcoholism, and/or drug abuse are among possible risk factors. Epilepsy is a chronic

neurological disease that affects 50 (WHO fact sheet, 2016) to 65 million people worldwide

(Thurman et al., 2011). Almost 80% of patients with epilepsy come from low- or middle-

income countries and approximately three fourths of patents with epilepsy do not receive

appropriate treatment. (WHO fact sheet, 2016). Even in highly developed countries with

common access to antiepileptic drugs, epileptic seizures are successfully treated in only 60-

70% of patients (Kwan and Brodie, 2000). Around 30-40% of patients with epilepsy don’t

respond to monotherapy and require a combination of 2 or 3 antiepileptic drugs

(polytherapy) (Kwan and Brodie, 2000). Uncontrolled epileptic seizures may lead to

increased mortality or developmental and cognitive dysfunctions and resulting in educational

or social disadvantages (Kwan and Brodie, 2001). Social stigma of unpredictable epileptic

seizures in patients with epilepsy may lead to decreased education and increased

unemployment or social isolation which result in mood disorders such as depression and

may lead to suicidal tendencies (Rai et al., 2012). In most cases epilepsy is incurable and

patients require continuing medication for the rest of their lives. Mortality in patients with

epilepsy is considerably higher than in the general population. Otherwise healthy patients


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with epilepsy are at the higher risk of sudden and unexpected death, a phenomenon called

SUDEP which stands for Sudden Unexpected Death in Epilepsy. SUDEP refers to the

sudden, unexpected, non-traumatic and nondrowning death in patients with epilepsy with

negative results of post-mortem examination in regards to an organic or toxicological cause

for death (Shorvon et al., 2011). Young adults 20-40 years old with poorly controlled or

intractable epilepsy are considered to be at the highest risk for SUDEP (Dlouhy et al., 2016).

The reasons for treatment resistance in epilepsy are still unknown but a variety of

neurological disorders such as cortical dysplasia, hippocampal sclerosis, ion channel and/or

receptor mutations, as well as infectious and autoimmunological processes are among

potential causes (Sanders, 2003). Despite the availability of three generations of antiepileptic

drugs, the effectiveness of epilepsy treatment has not significantly improved (Shorvon,

2009a,b). Considering the fact that 30-40% of patients with epilepsy require treatment with

more than one antiepileptic drug, achieving effectiveness and tolerability became the main

factor in choosing drug combinations. Taking under consideration individual molecular

mechanisms of action, antiepileptic drugs can be divided into four main groups: 1) voltage-

gated sodium, calcium and potassium channel modulators, 2) GABA-ergic

neurotransmission modulators that act through GABAA receptors or affect synthesis, uptake

and degradation of GABA, 3) those that decrease synaptic excitation trough action on

ionotropic glutamate receptors and 4) those that effect neurotransmitter systems through

presynaptic mechanisms, in particular glutamate release (Porter et al., 2012). There are

many institutions worldwide that conduct scientific searches to identify new synthetic and/or

natural origin compounds with potential antiseizure activity (Zolkowska et al., 2014; Desai et

al., 2015; Coleman et al., 2015). Despite some imperfections, animal seizure models enable

and allow for comprehensive and effective screening of a vast number of compounds to

detect substances possessing anticonvulsant activity (White, 2003). These animal seizure

models are often considered to be experimental models of certain seizure types in humans.

For example, the maximal electroshock seizure (MES) model is thought to be an

experimental model of tonic–clonic seizures and, to a certain extent, of partial convulsions

with or without secondary generalization in humans (Löscher et al., 1991). The MES model

is considered to be quite effective in identifying drugs that block generalized tonic–clonic

seizures in humans (Bialer and White, 2010). Moreover, compounds that were found to be

active in the MES test are generally efficacious in clinical trials. The MES test along with the

subcutaneous pentylenetetrazol (PTZ) model remain the “gold standards”; allowing fast

selection of a great number of compound candidates with potential antiseizure activity

(Rogawski, 2006). In the MES test in mice, seizures are induced with an alternating current

of the fixed intensity 25 mA, voltage 500 V, frequency of 50 Hz and duration of 0.2 seconds.

An electric impulse generated by a rodent shocker generator is applied via ear-clip

electrodes and results in tonic hind limb extension immediately after stimulation. The

protective action of antiepileptic drugs and compounds with antiseziure activity is manifested

as the increased number of animals protected from tonic hind limb extension after electric

stimulation. The maximal electroshock seizure threshold test (MEST) is used to evaluate the

antiseizure activity of compounds that are not considered antiepileptic. In the MEST the

median current strength value (CS50 median current strength in mA) predicted to produce

maximal seizure activity (tonic hind limb extension) in 50% of animals tested is established.

CS50 values of the tested compound are compared to CS50 values of the vehicle-treated

control animals. Increases or decreases to the CS50 value of the tested compound when

compared to the control group suggest antiseizure or proconvulsant activity respectively.


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The electroconvulsive threshold in the MEST test was calculated according to the log-probit

method of Litchfield and Wilcoxon (1949). The protective activity of a tested compound with

suspected antseizure activity in the MES test in mice is expressed as the protective median

effective dose (ED50) that shows drug potency. This ED50 value represents a dose of the

antiseizure drug (in mg/kg) predicted to protect 50% of mice tested against MES-induced

extension of the hind limbs and is also calculated according to log-probit method of Litchfield

and Wilcoxon (1949). The log-probit method compares quantitative (increases or decreases

in the tested drug ED50 value) and qualitative (changes of ED50 value of tested drug vs ED50

value in control group) changes in the potency of a tested compound. Pharmacoresitance in

epilepsy stimulates scientist worldwide to introduce new animal models of seizures and

epilepsy and explore new compounds and drugs combinations. Pharmacological profiles and

side effects assessments of newly synthesized compounds that affect the central nervous

system (CNS) are important steps in the search for substances with potential therapeutic

activity. Discovering new treatment options for pharmacoresistant epilepsy and status

epilepticus is the main focus of my research. My main interests include compounds with

potential antiepileptic activity that could be used as add-on drugs in polytherapy with

classical or new generation antiepileptic drugs (AEDs). Ideal add-on drugs should exhibit a

clean pharmodynamic and phamcokinetic profile to minimize possible drug interactions and

side effect profiles of combination therapy. For that reason, the antieseizure activity of

selected succinimide derivatives, nitric oxide (NO) synthase inhibitors (7-nitroindazole and

NG-nitro-L-arginine), modafinil and MPEP (a selective antagonist for the glutamate

metabotropic receptor subtype mGluR5) were extensively explored in my scientific

achievements.

1. Zolkowska D, Kondrat-Wrobel MW, Florek-Luszczki M, Luszczki JJ. Influence of

MPEP (a selective mGluR5 antagonist) on the anticonvulsant action of novel antiepileptic

drugs against maximal electroshock-induced seizures in mice. Prog Neuropsychopharmacol

Biol Psychiatry. 2016;65:172-178.

Glutamate is one of the main excitatory neurotransmitters in the CNS. Glutamate is

responsible for the majority of excitatory neurotransmission in mammalian brain through pre-

and postsynaptic metabotropic G-protein-coupled receptors and three types of ionotropic

receptors: presynaptic AMPA and NMDA and pre- and postsynaptic kainate receptors

(Barker-Haliski and White, 2015). Imbalance between excitatory glutamergic

neurotransmission and inhibitory GABA-ergic effects could cause neurons to destabilize and

result in epileptic seizure (Nadler 2012). Much attention has been paid in recent years to

metabotropic glutamate receptor modulators as more promising than those acting directly

through ionotropic AMPA and NMDA receptors (Pin and Acher, 2002). The anticonvulsant

activity of 2-methyl-6-(phenylethynyl)pyridine (MPEP) – selective antagonist of mGluR5

subtype metabotropic glutamate receptors had been previously reported in a variety of

experimental seizure models (MES, 6Hz, sound-induce seizures in DBA/2 mice) (Barton et

al., 2003). Upregulation of mGluR5 has been observed in the hippocampus of patients with

complex partial seizures (Notenboom et al., 2006). Previous studies demonstrated that

MPEP significantly increased the electroconvulsive threshold in the MEST model in mice but

had no effect on the anticonvulsant activity of conventional AEDs in the subprotective dose

(Zadrozniak et al., 2004). By contrast, MPEP combined with subprotective doses of

valproate (VPA) and phenobarbital (PB) decreased behavioral seizures and the EEG


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afterdischarge duration in amygdala-kindled seizures in rats (Borowicz et al., 2009). Based

on the information cited above, the effects of MPEP on the anticonvulsant activity of selected

new generation AEDs lamotrigine (LTG), oxcarbazepine (OXC), pregabalin (PGB) and

topiramate (TPM) in the MES test in mice became the main topic of my study. Noteworthy,

selected AEDs belong to the first-line of treatment (LTG, OXC) or add-on drugs in adult

patients that require more than one drug for treatment of tonic–clonic seizures (National

Clinical Guideline Centre, 2012). PGB is most often prescribed as an add-on therapy in

adults with partial onset seizures with or without secondary generalization when the drug of

first choice is ineffective (European Medicines Agency, 2009). In the study, the effects of

tested compounds on seizure thresholds in MEST test and ED50 values in MES seizure

models were evaluated. In addition, side effects of MPEP and combinations with the AEDs

were investigated using the step-through passive avoidance task (long-term memory

impairment), chimney test (motor coordination impairment) and grip strength test (muscular

strength impairment). Total brain concentrations of tested AEDs and their combinations with

MPEP (at the doses that corresponded to their ED50 values from the MES test) were

measured in homogenized brains to ascertain pharmacokinetic (PK) interactions. In the

beginning, the effect of MPEP on the seizure threshold was measured in MEST test and

subthreshold CS50 value was calculated for further use in combinations with AEDs.

Intraperitoneal administration of MPEP at the doses of 1.5 and 2 mg/kg significantly

increased the threshold for electrocolnvulsions in MEST test, but the dose of 1 mg/kg was

ineffective. On the other hand, MPEP in the MES test significantly increased the

anticonvulsant activity of PGB and TPM but didn’t affect the action of LTG and OXC.

Furthermore, none of the MPEP combinations with AEDs impaired memory, motor

coordination or muscle strength of tested animals. Coadminstration of MPEP (1 mg/kg) with

PGB did not change the total brain concentration of PGB as determined by high pressure

liquid chromatography (HPLC). Similarly, as determined by the fluorescence polarization

immunoassay, the concentration of TPM in homogenized brains was not significantly

affected by the combination with MPEP (1 mg/kg). Summarized, MPEP increased the

anticonvulsant activity of PGB and TPM without inducing unfavorable side effects. PK

studies suggested that favorable combinations of MPEP with PGB and TPM are

pharmacodynamic in nature and are worth exploring in further clinical studies.

2. Zolkowska D, Andres-Mach M, Prisinzano TE, Baumann MH, Luszczki JJ. Modafinil

and its metabolites enhance the anticonvulsant action of classical antiepileptic drugs in the

mouse maximal electroshock-induced seizure model. Psychopharmacology (Berl).

2015;232(14):2463-2479.

Modafinil (2-[(Diphenylmethyl) sulfinyl] acetamide, Provigil) is a wakefulness promoting

agent approved by FDA for the treatment of excessive sleepiness in narcolepsy, shift work

sleep problems, and obstructive sleep apnea (Minzenberg and Carter 2008). Modafinil binds

and moderately inhibits DAT and norepinephrine transporters but in the clinically relevant

dose (200 mg) is able to substantially inhibit both catecholamine transporters (Minzenberg

and Carter 2008). Modafinil is believed to affect a variety of central neurotransmitter

systems. Modafinil is reported to elevate extracellular levels of norepinephrine, serotonin,

histamine and glutamate; to reduce extracellular GABA levels by inhibiting GABA release;

and to activate α1 adrenergic receptors. Effects described above are particularly prominent

in the neocortex and generally less potent or minimal in the subcortical area. Modafinil’s

effects on dopamine and norepinephrine release appear to be primary while its influence on


8

the levels of other neurotransmitters (serotonin, GABA, glutamate, orexin and histamine)

seems to be secondary to the catecholamine effects (Minzenberg and Carter 2008).

Modafinil exerts antiepileptic effects despite reports that the drug reduces GABA activated

currents and extracellular GABA levels in different brain regions (Chen et al., 2007). In the

study of Chen et al. (2007) modafinil reduced seizure activity in the MES test and the PTZ-

kindling model. Moreover, the sulfone and acid metabolites of modafinil are reported to exert

anticonvulsant activity in the MES model (Chatterjie et al., 2004). In the past decade, there

has been a marked increase in prescription of modafinil for a variety of indications not

approved by the FDA (Peñaloza et al., 2013). Modafinil is often prescribed to epileptic

patients that suffer from fatigue, excessive daytime sleepiness and cognitive dysfunction

caused by antiepileptic treatments (Artsy et al., 2012).

Increasing the use of modafinil in patients with epilepsy inspired my next study, where I

explored the effects of modafinil and its two metabolites, diphenylmethylsufonylacetamide

(i.e., sulfone metabolite) and diphenylmethylthioacetic acid (i.e., acid metabolite), on the

anticonvulsive activity of four classical AEDs: carbamazepine (CBZ), phenobarbital (PB),

phenytoin (PHT), and valproate (VPA) in the mouse MES-induced seizure model. As

modafinil is a DAT blocker, the anticonvulsant effects to the prototypical DAT blocker1-[2-

[bis(4- fluorophenyl)methoxy]ethyl]-4-(3-phenylpropyl)piperazine (GBR 12909) was

compared. Similar to the previous study, the threshold for electroconvulsions of all tested

substances was explored. Furthermore, anticonvulsant activity in the MES test was

assessed and the ED50 value was calculated. In addition, the side effects of modafinil, its

metabolites and GBR 12909 and their combination with classical AEDs in the chimney test,

step-through passive avoidance task and grip-strength test were evaluated. A PK

assessment of the total brain concentrations of the AEDs for their combinations with

modafinil, its metabolites and GBR 12909 was performed. Intraperitoneal administration of

75 mg/kg of modafinil or its metabolites significantly elevated the threshold for

electroconvulsions in mice in the MEST test. Additionally, intraperitoneal administration of

GBR 12909 at the dose of 50 mg/kg significantly elevated the threshold for

electroconvulsions in mice. It’s worth mentioning that the dose of modafinil used to elevate

the threshold for electroconvulsions (75 mg/kg) is considerably below the dose necessary to

stimulate robust motor activity. On the other hand, the doses of GBR 12909 affecting the

electroconvulsive threshold are in the range of motor stimulant doses. According to Paterson

et al. (2010) 150 mg/kg of modafinil and 15 mg/kg of GBR 12909 elicit equivalent locomotor

activation in mice. Thus, it seems that the anticonvulsant effects of modafinil may not be

related solely to its effects on the central dopaminergic activity. The subthreshold dose of

modafinil and its metabolites (50mg/kg) significantly elevated the anticonvulsant activity of

CBZ, PHT, and VPA but did not change the action of PB. Modafinil and its sulfone

metabolite at the dose of 25 mg/kg enhanced only the anticonvulsant activity of VPA. GBR

12909 at the subthreshold dose of 25 mg/kg significantly enhanced the anticonvulsant

activity of CBZ, PB, PHT and VPA; being ineffective at the dose of 12.5 mg/kg. Modafinil and

its metabolites (50 mg/kg) did not alter the total brain concentration of PB and VPA but

elevated CBZ and PHT concentrations. According to results presented by Robertson and

Hellriegel (2003), clinically significant interactions of modafinil with other drugs are most

likely connected to effects on two hepatic enzymes: CYP3A4/5 (most prevalent human CYP

enzyme) and CYP2C9. Studies using human liver microsomes revealed that modafinil can

cause weak inhibition of CYP3A4/5. CBZ is primarily metabolized by CYP3A4, which is why

brain levels of CBZ may be increased in combination with modafinil or its metabolites.


9

Modafinil and its sulfone metabolite inhibit CYP2C9 in human hepatocytes and 2C isoforms

are major catalysts of PHT metabolism in humans (Cuttle et al., 2000; Robertson and

Hellriegel 2003). The total brain concentration of CBZ, PB, PHT and VPA was not affected

by their combinations with GBR 12909. Combinations of modafinil, its metabolites or GBR

12909 with AEDs did not induce adverse effects in tested animals. Collectively, our study

was first to report the positive effect of modafinil and its metabolites on seizure threshold in

experimental animals. My study suggests that the combination of modafinil with classical

AEDs does not increase the risk for seizures but may be a beneficial therapeutic

combination with no exacerbation of side effects. Moreover, a retrospective analysis of

clinical data demonstrated that epileptic patients taking modafinil for over 10 years had no

exacerbation of seizures (Artsy et al., 2012). Use of modafinil seems to be a safe solution in

patients with epilepsy suffering from sleepiness, fatigue and cognitive dysfunction caused by

antiepileptic treatments.

3. Luszczki JJ, Jaskolska A, Dworzanski W, Zolkowska D. 7-nitroindazole, but not NG-

nitro-L-arginine, enhances the anticonvulsant activity of pregabalin in the mouse maximal

electroshock-induced seizure model. Pharmacol Rep. 2011;63:169-75.

Nitric oxide (NO) is one of the most active gaseous molecules in living organisms. Results of

in vitro studies revealed the strong neurotransmitter/neuromodulator-like properties of NO in

the CNS. Moreover, it has been accepted that NO plays a critical role in many physiological

and pathological processes in the CNS (Garthwaite, 1991). NO is synthesized from the L-

arginine by NO synthase (NOS) which exists in three distinct isoforms: endothelial (eNOS),

neuronal (nNOS) and inducible (iNOS) (Ferraro and Sardo, 2004). NO plays an important

role in the pathophysiology of epilepsy taking part in the genesis and spreading of

epileptiform activity (Przegalinski et al., 1996). As suggested by previous studies, the role of

NO in convulsive phenomena may differ, producing both anti- and pro-convulsant effects

depending on the experimental epilepsy model (Ferraro and Sardo, 2004). Molsidomine (NO

donor), L-arginine (natural NO precursor), NG-nitro-L-arginine (NNA– a non-selective NOS

inhibitor, decreasing the activity of eNOS and nNOS) and 7-nitroindazole (7NI, preferential

inhibitor of nNOS activity) are considered NO modulators in the CNS (Moncada and Higgs,

1995). Results of the previous study indicated that PGB exhibits anticonvulsant activity in the

MES and PTZ seizure models (Vartanian et al., 2006). Considering the aforementioned results

I investigated the effects of 7NI and NNA on the anticonvulsant action of PGB in the MES

seizure model in mice. 7NI administered intraperitoneally at the dose of 50 mg/kg

significantly enhanced the anticonvulsant action of PGB and did not impaire motor

coordination, memory or muscle strength of tested animals. However, NNA at the dose of 40

mg/kg did not affect the anticonvulsant activity of PGB. Total brain concentrations of PGB in

combination with 7NI were not tested because of the small likelihood of PK interactions.

Furthermore, the previously mentioned results indicated that 7NI administered at the dose of

150 mg/kg had no impact on total brain concentrations of CBZ, PHT, PB and VPA in mice

(Luszczki, 2006). In conclusion, the combination of 7NI with PGB deserves more attention

and further studies in other seizure models because of its favorable effects in terms of

increased anticonvulsant activity and lack of significant acute adverse effects; such

combination may be an important treatment option for patients with intractable epilepsy.


10

4. Zolkowska D, Kominek M, Florek-Luszczki M, Kocharov SL, Luszczki JJ. Effects of

N-(morpholinomethyl)-p-isopropoxyphenylsuccinimide on the protective action of different

classical antiepileptic drugs against maximal electroshock-induced tonic seizures in mice.

Pharmacol Rep. 2013;65:389-398.

5. Luszczki JJ, Kominek M, Florek-Luszczki M, Tchaytchian D.A, Kocharov SL,

Zolkowska D. Influence of N-hydroxymethyl-p-isopropoxyphenylsuccinimide on the

anticonvulsant action of different classical antiepileptic drugs in the mouse maximal

electroshock-induced seizure model. Epilepsy Res. 2012;100:27-36.

6. Luszczki JJ, Cioczek JD, Kocharov SL, Andres-Mach M, Kominek M, Zolkowska D.

Effects of three N-(carboxyanilinomethyl) derivatives of p-isopropoxyphenylsuccinimide on

the anticonvulsant action of carbamazepine, phenobarbital, phenytoin and valproate in the

mouse maximal electroshock-induced seizure model. Eur J Pharmacol. 2010;648:74-79.

The anticonvulsive activity of succinimide derivatives were confirmed in many studies using

a variety of animal seizure models (Zejc et al., 1990; Kaminski and Obniska, 2008). At the

current stage of experimental development, little is known about the molecular mechanism of

observed anticonvulsive activity of succinimide derivatives. Based on in vitro radioligand

binding studies, it has been reported that some spirosuccinimide derivatives are able to

block voltage-gated sodium channels with higher affinity than PHT (Obniska et al., 2010). In

three consecutive publications I examined the effects of newly synthesized succinimide

derivatives: N-(morpholinomethyl)-p-isopropoxy-phenylsuccinimide (MMIPPS), N-

hydroxymethyl-p-isopropoxyphenylsuccinimide (HMIPPS), N-(ortho-carbox-yanilinomethyl)-

p-isopropoxyphenylsuccinimide (o-CAMIPPS), N-(meta-carboxyanilinomethyl)-p-

isopropoxyphenylsuccinimide (m-CAMIPPS), and N-(para-carboxyanilinomethyl)-p-

isopropoxyphenylsuccinimide (p-CAMIPPS) on the anticonvulsant activity of selected AEDs

in the MES seizure model in mice. Intraperitoneal administration of o-CAMIPPS, m-

CAMIPPS, and p-CAMIPPS at the doses of 75-150 mg/kg significantly elevated the

threshold for electroconvulsions in the MEST test in mice, but were ineffective at the dose of

37.5 mg/kg. Moreover, o-CAMIPPS in the subthreshold dose significantly reduced the

anticonvulsant activity of CBZ, but not that of PHT, PB and VPA in the MES test in mice.

Succinimide derivatives m-CAMIPPS and p-CAMIPPS were unable to affect the

anticonvulsant activity of CBZ, PHT, PB and VPA with the MES test. Results of the PK

studies revealed that o-CAMIPPS significantly increased total brain concentrations of CBZ in

mice. The reduced anticonvulsant action of CBZ in combination with o-CAMIPPS with

concurrent increase of the total brain concentration of CBZ may suggest the antagonistic

interaction between tested drugs. Preclinical studies of the combinations of m-CAMIPPS or

p-CAMIPPS with classical AEDs reveal a neutral interaction. Obtained results confirm that

chemical substitution of the carboxyanilinomethyl group to the ortho-, meta- or para- location

of p-isopropoxyphenylsuccinimide did not create compounds that presented beneficial

anticonvulsant profiles in combination with classical AEDs. In the consecutive study I

examined the effects of HMIPPS on the anticonvulsive action of CBZ, PHT, PB and VPA in

the MES seizure model in mice. HMIPPS administered intraperitoneally at the dose of 100

mg/kg, but not those of 50 mg/kg and lower, significantly elevated the threshold for

electroconvulsions assessed in the MEST test in mice. HMIPPS at the subthreshold dose of

50 mg/kg significantly enhanced the anticonvulsant activity of PB and VPA but did not affect

the activity of CBZ and PHT in the MES seizure model. HMIPPS at the subthreshold dose of

25 mg/kg significantly enhanced the anticonvulsant action of VPA but not that of other tested


11

AEDs. PK studies revealed that HMIPPS did not alter total brain concentrations of PB and

VPA. HMIPPS (50 mg/kg) in combinations with classical AEDs did not impair motor

coordination, long-term memory and muscular strength in tested animals. Based on the

obtained results, one may conclude that the combination of HMIPPS with PB and VPA

presents a clinically beneficial increase in anticonvulsant activity without PK interactions or

exacerbation of side effects. Such combinations are worthy of consideration for further

experimental studies and possible clinical use. The combinations of HMIPPS with CBZ and

PHT, however, are neutral from a preclinical viewpoint. In my next study with succinimide

derivatives I examined the effects of MMIPPS on the on the anticonvulsive action of classical

AEDs CBZ, PB, PHT and VPA in the MES seizure model in mice. Intraperitoneal

administration of MMIPPS at the dose of 100 mg/kg significantly elevated the threshold for

electroconvulsions in MEST test. MMIPPS at the subthreshold dose (50 mg/kg) significantly

enhanced the anticonvulsant activity of PB and VPA, but not that of CBZ or PHT, in the MES

test. PK studies demonstrated that MMIPPS (50 mg/kg) did not alter total brain

concentrations of PB, but significantly increased total brain concentrations of VPA in mice.

None of the drug combinations greatly affected motor coordination, long-term memory or

muscular strength in tested animals. In conclusion, the results obtained in this study suggest

that the combination of MMIPPS with AEDs, especially with PB, may be a beneficial

therapeutic option for patients with drug–resistant epilepsy.

Summary:

In publications that are a part of my scientific achievement I demonstrated that screening

new compounds with potential antiseizure activity is the key factor in identification of new

treatment options for 30-40% of patients with epilepsy that despite polytherapy suffer from

uncontrollable seizures attacks.

1) MPEP, a selective antagonist of mGluR5 subtype metabotropic glutamate receptors in

subthreshold doses enhanced the anticonvulsant activity of PGB and TMP without

exacerbation of side effects. Favorable pharmacodynamic in nature interactions of MPEP

with PGB and TPM are worth exploring further in clinical studies. Experimental results

suggest that co-administration of AEDs with antagonists of mGluR5 subtype metabotropic

glutamate receptors may be promising combination therapies.

2) Modafinil may be a safe and beneficial addition to the therapy with classical AEDs that

doesn’t decrease tolerability. Augmentation of antiepileptic effects combined with the help

against sleepiness, fatigue and cognitive dysfunction caused by antiepileptic treatments may

be especially important for working patients with epilepsy.

3) My studies confirmed earlier observations regarding the important role of NO in epilepsy.

Combination of 7NI with PGB deserves attention and further studies because of its favorable

effects in terms of increased anticonvulsant activity and lack of significant adverse effects.

4) Results of studies with the effects of newly synthesized p-isopropoxy-phenylsuccinimide

derivatives suggested that N-(morpholinomethyl)- (MMIPPS) and N-hydroxymethyl-

(HMIPPS) derivatives enhanced the anticonvulsive activity of PB and VPA without

exacerbation of side effects. Obtained results confirm that the chemical substitution of the


12

carboxyanilinomethyl group to the ortho-, meta- or para- location of p-

isopropoxyphenylsuccinimide did not create compounds that in combination with classical

AEDs presented a beneficial anticonvulsant profile.

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Desai R, Savechenkov PY, Zolkowska D, Ge RL, Rogawski MA, Bruzik KS, Forman SA, Raines DE, Miller KW. Contrasting actions of a convulsant barbiturate and its anticonvulsant enantiomer on the α1 β3 γ2L GABAA receptor account for their in vivo effects. J Physiol. 2015;593(22):4943-4961.

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Pharmacol. 2004;56:595-598.


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Zejc A, Obniska J, Wilimowski M, Rutkowska M, Witkowska M, Barczynska J, Kedzierska-

Gozdzik L, Wojewodzki W, Orzechowska-Juzwenko K, Plawiak T. Synthesis and

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Zolkowska D, Dhir A, Krishnan K, Covey DF, Rogawski MA. Anticonvulsant potencies of the enantiomers of the neurosteroids androsterone and etiocholanolone exceed those of the natural forms. Psychopharmacology (Berl). 2014;231:3325-3332.

5. Discussion of the other scientific achievements.

Studying and applying new forms of therapy fascinated me from the early age and inspired

me to pursue medical school. In 1994 I finished medical school with excellent results and

started an internship at the Provincial Hospital (Wojewodzki Szpital Zespolony im. Jana

Bozego) in Lublin, Poland. While volunteering I received the doctoral fellowship in the Ph.D.

program of the Department of Pharmacology and Toxicology at the Medical University in

Lublin, Poland. I was honored to work under the leadership of Prof. dr h. c. mult. Zdzisław

Kleinrok alongside Prof Maria Sieklucka-Dziuba, Prof. Stanisław Jerzy Czuczwar and Prof.

Grażyna Rajtar-Cynke.

My research was exploring newly synthesized compounds with suspected action on the

CNS; research that was published in three different publications. My studies revealed that N-

[4-phenyl(2-pyrimidinyl)-1-piperazinyl]alkyl-(hydroxyalkyl)-3,4-pyridinedicarboximides did not

exhibit antiepileptic or anxiolytic activity. Obtained results suggested that the substitution of

pyridine group in position 2 of 3,4- pyridinedicarboximides with other groups did not

significantly change their pharmacological profile. Some of the tested imides displayed

depressive action on the CNS expressed as inhibition of spontaneous locomotor activity

(publication 1). In most cases heterocyclic compound derivatives induced moderate toxicity

and doses equal to one tenth of their LD50 value did not cause neurotoxic effects or impair

motor coordination in a rotarod test. Tested heterocyclic compound derivatives significantly

inhibited spontaneous locomotor activity and possessed strong analgesic activity especially

evident in the “writhing test” in mice (publication 2). The aim of the study with aminoalkanolic

derivatives of xanthones was to assess their effects on the CNS. The (R)-2-N-methylamino-

1-butanol derivative of 7-chloro-2-methylxanthone expressed the most interesting

pharmacological profile. Anticonvulsant activity of the aforementioned compound was

observed in the MES and PTZ-induced seizures tests. Furthermore, this compound did not

affect GABA levels in the brain, had a relatively low toxicity and did not exhibit a neurotoxic

effect (publication 3).

1. Rajtar G, Żółkowska D, Kleinrok Z, Marona H. Pharmacological properties of some

xanthone derivatives. Acta Pol Pharm. 1999;56(4):311-318.

2. Śladowska H, Bodetko M, Sieklucka-Dziuba M, Rajtar G, Żółkowska D, Kleinrok Z.

Transformation of some pyrido[2,3-d]pyrimidine derivatives into other di- and

triheterocyclic systems. Farmaco. 1997;52(11):657-662.

3. Śladowska H, Potoczek J, Rajtar G, Sieklucka-Dziuba M, Żółkowska D, Kleinrok Z.

Investigations on the synthesis and properties of some N-[4-phenyl(2-pyrimidinyl)-1-

piperazinyl]alkyl (hydroxyalkyl)-3,4-pyridinedicarboximides. Acta Pol Pharm. 1996

;53(6):417-424.


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The aim of my next project was to examine the effects of xanthone derivatives and classical

AEDs (VPA, PB, DZP, clonazepam [CZP], CBZ and PHT) on platelet function in rats. In

addition, I studied the effects of DZP, CZP and peripheral benzodiazepine receptor

antagonist PK 11195 on the neutrophil’s function. Screening of the twelve aminoalkanolic

derivatives of xanthone revealed five compounds with a range of potency that inhibited

thrombin-induced platelet aggregation. Most of the tested compounds dose-dependently

inhibited arachidonic acid or adenosine-5'-diphosphate induced platelet aggregation.

Experiments exploring the effects of conventional AEDs on the platelet function revealed that

only DZP, and to a smaller extent CZP, impaired platelet aggregation, malondialdehyde

synthesis and increased free intracellular calcium levels in stimulated platelets. The

described effects are at least partially due to the influence of tested compounds on the

arachidonic acid pathways and inhibition of proaggregatory prostanoids synthesis. DZP,

CZP and PK 11195 alone did not simulate resting platelets or modified neutrophils response.

Moreover, DZP and CZP exerted an inhibitory effect on reactive oxygen species (ROS)

produced by formyl methionyl leucyl phenylalanine (fMLP) activated neutrophils. The

antiplatelet effects of DZP and CZP combined with the inhibitory action on neutrophils ROS

production may have both positive and negative practical aspect (in patients with

coagulopathy). DZP and CZP inhibitory actions on fMLP induced ROS production in

neutrophils may be a significant factor in cell protection against free radicals. On the other

hand, prolonged use of benzodiazepines may reduce cell mediated immunity. Based on the

aforementioned results two interesting publications in journals with IF were created.This

research project was carried out under the tutelage of Prof. dr h. c. mult. Zdzisław Kleinrok

and direct supervision of Prof. Grażyna Rajtar-Cynke resulted in my doctoral thesis titled:

„Effect of diazepam and clonazepam on rat’s platelets and neutrophils function” and was

defended in 1999 at the Medical University in Lublin.

1. Rajtar G, Żółkowska D, Kleinrok Z. Effect of diazepam and clonazepam on the

function of isolated rat platelet and neutrophil. Med Sci Monit. 2002;8(4):PI37-PI44.

2. Rajtar G, Żółkowska D, Czechowska G, Kleinrok Z. Effects of antiepileptic drugs on

rat platelet aggregation: ex vivo and in vitro study. Epilepsy Res. 2001;43(1):59-66.

In the next project the effects of the NO on seizure susceptibility in the seizure model

induced by potassium channel blocker 4-aminopyridine (4-AP) in mice was tested. It has

been suggested that NO synthesis and release may contribute to the generation of seizure

activity in response to excessive glutamate-mediated excitation induced by 4-AP. The

effects of NNA (an inhibitor of NO synthase) and two agents elevating NO (L-arginine and

molsidomine) were tested in the 4-AP seizure model. Results of the study suggest lack of

the NO involvement in the mechanisms underlying the development of seizures and

lethality induced by 4-AP. Proconvulsant action of NNA and the potentiation of this action

by molsidomine are most probably due to mechanisms unrelated to NO synthesis.

Moreover, endogenous histamine is also involved in the regulation of seizure susceptibility.

The following study investigated the influence of L-histidine on the protective effects of CBZ

and PHT in the MES seizure model in mice. Subthreshold doses of L-histidine (500mg/kg)

enhanced the anticonvulsant activity of both tested AEDs in the MES seizure model.

Importantly, the combinations of tested AEDs with L-histidine were not associated with

augmentation of adverse effects. Results of this project were published in 3 publications.


17

1. Kamiński RM, Żółkowska D, Kozicka M, Kleinrok Z, Czuczwar SJ. L-histidine is a

beneficial adjuvant for antiepileptic drugs against maximal electroshock-induced

seizures in mice. Amino Acids. 2004;26(1):85-89.

2. Tutka P, Młynarczyk M, Żółkowska D, Kleinrok Z, Wielosz M, Czuczwar SJ. Nitric

oxide and convulsions in 4-aminopyridine-treated mice. Eur J Pharmacol.

2002;437:47-53.

3. Tutka P, Młynarczyk M, Żółkowska D, Kleinrok Z. NG-nitro-L-arginine sensitizes

mice to 4-aminopyridine-induced seizures. Acta Physiol Hung. 1996;84(4):471-472.

After finishing my doctoral fellowship I was employed at the Department of Hygiene, Medical

University in Lublin as a research associate and then as assistant professor. I continued my

research focusing on the variety of compounds affecting the CNS, including pesticides and

heavy metals (cadmium, lead). Shortly after obtaining my Ph.D. degree I received the

prestigious Fogarty Foundation fellowship at the National Institute on Drug Abuse, National

Institutes of Health, Baltimore, MD, USA. From 2002 until 2004 I was working as a Visiting

Fellow at the Integrative Neuroscience Section, Behavioral Neuroscience Branch, National

Institute on Drug Abuse, Baltimore, MD, USA where I was involved in exploring the role of

dynorphin/NMDA interaction in the development of opioid–induced hyperalgesia and chronic

pain and the neurotoxic effects of dynorphin peptides in the spinal cord. In 2004 my research

efforts were awarded by the travel grant, Fellows Award for Research Excellence (FARE)

that allowed me to present my data at the Society of Neuroscience meeting in New Orleans,

LA, USA. Results of my study were published in the renowned Journal of Proteome

Research with an IF=5.151.

1. Woods AS, Kamiński R, Oz M, Wang Y, Hauser K, Goody R, Wang HY, Jackson SN,

Zeitz P, Zeitz KP, Zolkowska D, Schepers R, Nold M, Danielson J, Gräslund A,

Vukojevic V, Bakalkin G, Basbaum A, Shippenberg T. Decoy peptides that bind

dynorphin noncovalently prevent NMDA receptor-mediated neurotoxicity. J Proteome

Res. 2006;5(4):1017-1023.

While working as a Visiting Fellow at the Clinical Psychopharmacology Section, Medications

Discovery Branch, National Institute on Drug Abuse, Baltimore, MD, USA from 2002 until

2004 I was involved in exploring the effect of clinically used medication, addictive drugs,

ligands of serotonin and dopamine receptors on the serotonin levels in blood as well as

levels of other neurotransmitters in the rat brain. Results of my work were published in 6

papers. My publication exploring the wakefulness-promoting agent modafinil was cited by

the prestigious Faculty of 1000 Biology and obtained F1000 Factor 6.0

(http://f1000.com/prime/1148208).

1. Baumann MH, Williams Z, Zolkowska D, Rothman RB. Serotonin (5-HT) precursor

loading with 5-hydroxy-l-tryptophan (5-HTP) reduces locomotor activation produced

by (+)-amphetamine in the rat. Drug Alcohol Depend. 2011;114:147-152.

2. Baumann MH, Zolkowska D, Kim I, Scheidweiler KB, Rothman RB, Huestis MA.

Effects of dose and route of administration on pharmacokinetics of (±)-3,4-

methylenedioxymethamphetamine (MDMA) in the rat. Drug Metabolism and

Disposition. 2009; 37:2163-2170.

http://f1000.com/prime/1148208


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3. Zolkowska D, Jain R, Rothman RB, Partilla JS, Roth BL, Setola V, Prisinzano TE,

Baumann MH. Evidence for the involvement of dopamine transporters in behavioral

stimulant effects of modafinil. J Pharmacol Exp Ther. 2009; 329:738-746.

4. Zolkowska D, Baumann MH, Rothman RB. Chronic fenfluramine administration

increases plasma serotonin (5-hydroxytryptamine) to nontoxic levels. J Pharmacol

Exp Ther. 2008;324(2):791-797.

5. Rothman RB, Zolkowska D, Baumann MH. Serotonin (5-HT) transporter ligands

affect plasma 5-HT in rats: Implications for medication development. Ann N Y Acad

Sci. 2008;1139: 268–284.

6. Zolkowska D, Rothman RB, Baumann MH. Amphetamine analogs increase plasma

serotonin: implications for cardiac and pulmonary disease. J Pharmacol Exp Ther.

2006;318(2):604-610.

After finishing my postdoctoral training I continued my research as a Project Scientist at the

Department of Neurology, School of Medicine University of California, Davis, Sacramento,

CA, USA. In the same time I actively pursued collaboration with Prof. Jarogniew Luszczki

from the Medical University in Lublin, Poland and Dr Baumann from the Medicinal Chemistry

Section, Molecular Targets and Medications Discovery Branch, National Institute on Drug

Abuse, Baltimore, MD, USA where I hold the status of Guest Researcher since 2012.

My research is focused on the investigation of new strategies for the treatment of epileptic

seizures, epilepsy and status epilepticus in children and adults utilizing EEG analysis and

behavioral animal models. Moreover, I am one of the lead researchers working on seizure-

inducing agents listed by the WHO as chemical threat agents

(http://counteract.ucdavis.edu/project-core/project-1-prevention-termination-seizures).

My work exploring the effects of neurosteroids in treatment of status epilepticus resulted in

co-authorship in a patent application filled in 2012 (Rogawski MA, Zolkowska D.

Anticonvulsant Activity of Steroids U.S. Patent Application 61/732,252 filed Nov 30, 2012).

The application was licensed by the University of California and is currently in development

by a commercial entity. In 2012 I received an Innovative Developmental Award (IDA) grant

funded by the UC Davis Academic Federation Research Grant Program. My project titled

“Identification of novel therapeutic approaches for TETS intoxication” obtained valuable

preliminary data on the seizure inducing compound known as

tetramethylenedisulfotetramine (tetramine, TETS). TETS is a highly toxic rodenticide that

was used in the past as a rat poison. TETS is odorless, tasteless and is 100 times more

toxic then cyanide. It easily dissolves in water and is very stable in the environment and the

food chain. TETS, similar to picrotoxin, is a strong GABAA receptor blocker. Despite

comparable activity on the GABAA receptor between TETS and picrotoxin, TETS’ convulsant

activity is 30 to100 times stronger than that of picrotoxin with similar effects on lethality in

exposed animals. Currently the basis for the high in vivo convulsant potency of this

compound is not well understood. Humans that are severely poisoned with TETS develop

treatment-resistant status epilepticus often followed by a coma and death. Current medical

countermeasures for acute TETS intoxication can prevent mortality but do not sufficiently

protect the CNS from persistent status epilepticus. As synthesis of TETS is relatively easy

and the lethal dose in humans is so miniscule (0.1 mg/kg), TETS is considered a very high

priority chemical threat agent. In 2012 the project titled “Novel anticonvulsant and

neuroprotective therapies for TETS and OP intoxication” received a grant from the NIH

Countermeasures Against Chemical Threats (CounterACT) Research Program and the UC



19

Davis CounterACT Center of Excellence, an integral part of the NIH Countermeasures

Against Chemical Threats Research Network, was formed. As a leading researcher of

Project 1 titled “Identification of treatments for chemical threat agent seizures” I continue my

research on TETS toxicity. While TETS intoxication in humans leads to treatment-resistant

status epilepticus, the experimental animal model produces generalized and rapidly lethal,

tonic-clonic convulsions typical of GABAA receptor antagonists like PTZ or picrotoxin. Based

on many years of scientific experience I developed and established a model of TETS-

induced status epilepticus in mice; a pivotal mile stone for the grant and the UC Davis

CounterACT Center of Excellence. Results of my research were presented at multiple

international scientific conferences: Society for Neuroscience meeting and American

Epilepsy Society, Annual CounterACT Network Research Symposium, Toxicology Annual

Meeting and my attendance was often funded by UC Davis Academic Federation Research

Travel grants. Based on the results of my research and collaborations within and beyond the

UC Davis CounterACT Center of Excellence a total of 11 papers were published in

renowned scientific journals.

1. Kazdoba TM, Hagerman RJ, Zolkowska D, Rogawski MA, Crawley JN.

Evaluation of the neuroactive steroid ganaxolone on social and repetitive

behaviors in the BTBR mouse model of autism. Psychopharmacology (Berl).

2016;233(2):309-23.

2. Desai R, Savechenkov PY, Zolkowska D, Ge RL, Rogawski MA, Bruzik KS,

Forman SA, Raines DE, Miller KW. Contrasting actions of a convulsant

barbiturate and its anticonvulsant enantiomer on the α1 β3 γ2L GABAA receptor

account for their in vivo effects. J Physiol. 2015;593(22):4943-61.

3. Zolkowska D, Dhir A, Krishnan K, Covey DF, Rogawski MA. Anticonvulsant

potencies of the enantiomers of the neurosteroids androsterone and

etiocholanolone exceed those of the natural forms. Psychopharmacology (Berl).

2014;231:3325-3332.

4. Vito ST, Banks CN, Austin AT, Inceoglu B, Bruun DA, Zolkowska D, Tancredic

DJ, Rogawski MA, Hammocka BD, Lein PL. Post-Exposure Administration of

Diazepam Combined with Soluble Epoxide Hydrolase Inhibition Stops Seizures

and Modulates Neuroinflammation in a Murine Model of Acute TETS Intoxication.

Toxicol Appl Pharmacol. 2014 Dec 1;281(2):185-194.

5. Coleman N, Nguyen HM, Cao Z, Brown BM, Jenkins DP, Zolkowska D, Chen

YJ, Tanaka BS, Goldin AL, Rogawski MA, Pessah IN, Wulff H. The Riluzole

Derivative 2-amino-6-trifluoromethylthio-benzothiazole (SKA-19), a mixed KCa2

activator and Na V blocker, is a potent novel anticonvulsant. Neurotherapeutics.

2015;12(1):234-249.

6. Inceoglu B, Zolkowska D, Yoo HJ, Wagner KM, Yang J, Hackett E, Hwang SH,

Lee KS, Rogawski MA, Morisseau C, Hammock BD. Epoxy fatty acids and

inhibition of the soluble epoxide hydrolase selectively modulate GABA mediated

neurotransmission to delay onset of seizures. PLoS One. 2013 Dec 11;8:e80922.

7. Rogawski MA, Loya CM, Reddy K, Zolkowska D, Lossin C. Neuroactive steroids

for the treatment of status epilepticus. Epilepsia. 2013;54 Suppl 6:93-98.

8. Bröer S, Zolkowska D, Gernert M, Rogawski MA. Proconvulsant Actions of

Intrahippocampal Botulinum Neurotoxin B in the Rat. Neuroscience.

2013;252:253.


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9. Dhir A, Zolkowska D, Rogawski MA. Seizure protection by intrapulmonary

delivery of midazolam in mice. Neuropharmacology. 2013;73C:425-431.

10. Zolkowska D, Banks CN, Dhir A, Inceoglu B, Sanborn JR, McCoy MR, Bruun DA,

Hammock BD, Lein PJ, Rogawski MA. Characterization of seizures induced by

acute and repeated exposure to tetramethylenedisulfotetramine. J Pharmacol

Exp Ther. 2012; 341:435-446.

11. Dhir A, Zolkowska D, Murphy R, Rogawski MA. Seizure protection by

intrapulmonary delivery of the propofol prodrug propofol hemisuccinate. J

Pharmacol Exp Ther. 2011;336: 215-222.

Moreover, beyond my scientific activities at UC Davis I continue to actively contribute to the

scientific collaborations exploring drugs and newly synthesized compounds with potential

anticonvulsant activity in animal seizure models with the Medical University in Lublin and the

Medicinal Chemistry Section, Molecular Targets and Medications Discovery Branch,

National Institute on Drug Abuse, Baltimore, MD USA. As a result of these studies 9 papers

were published, 6 of them are included as a part of my scientific achievement and additional

3 are presented below.

1. Andres-Mach M, Zolkowska D, Barcicka-Klosowska B, Haratym-Maj A, Florek-

Luszczki M, Luszczki JJ. Effect of ACEA-A selective cannabinoid CB1 receptor

agonist on the protective action of different antiepileptic drugs in the mouse

pentylenetetrazole-induced seizure model. Prog Neuropsychopharmacol Biol

Psychiatry. 2012;39:301-309.

2. Luszczki JJ, Jaskolska A, Dworzanski W, Zolkowska D. Effect of 7-nitroindazole and

NG-nitro-L-arginine on the protective action of clobazam in the maximal

electroshock-induced seizures in mice. Annales UMCS, Sectio DDD. 2010; XXIII,

N3:135-143.

3. Luszczki JJ, Misiuta-Krzesinska M, Wlaz A, Zwolinski J, Cioczek JD, Zolkowska D.

WIN 55,212-2 mesylate (a highly potent non-selective cannabinoid CB1 and CB2

receptor agonist) elevates the threshold for maximal electroshock-induced seizures

in mice. Journal of Pre-Clinical and Clinical Research. 2009;3:110-113.

6. Summary of the scientific achievement

My current scientific achivement contains 44 publications, 41 oryginal publications, 1 review

and 2 publications in the journals supplements.

Original publications: 41

Total Impact Factor 95.732

Total number of points MniSW 813.50

Citations number according to Web of Science

(without autocitation) 344

H-index 10

department of neurology, school of medicine, university of

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