Prof. PharmDr. Milan Nobilis, CSc.

Date and Place of Birth: 29th June 1956, Hradec Králové, Czechoslovakia

Present position

Contact information:
Faculty of Pharmacy, Heyrovského 1203, 500 05 Hradec Králové, Czech Republic
Tel: +420 495067309
E-mail: milan.nobilis@faf.cuni.cz

Research interests

  • Study of the fate of medicaments in organism using separation instrumental analytical and xenobiochemical methods
  • Development and validation of novel analytical and bioanalytical, achiral and chiral methods [SPE(LLE)-HPLC/UHPLC-PDA-FL-MS/MS)] for the determination of new potential drug and its phase I and phase II metabolites in various biomatrices.
  • Synthesis of new potential drug (drug candidate) and its metabolites.
    The synthetic standards of above-mentioned parent compounds and their metabolites were utilized in various studies: ADME (Absorption-Distribution-Metabolism-Excretion), study of interspecies and gender differences, the formation of drug metabolites at various hierarchic levels of organism (molecular, subcellular, cellular, organs, whole organism),  pharmacodynamic and toxicology studies and the identification of enzymes involved in the drug biotransformation. For example, two potential antineoplastics were studied as seen below:
    (1982-2015) benfluron and its derivatives: see publications [1-12, 14, 15, 17, 19, 22-24, 26, 28, 29, 34, 61, 71], (1998-2015) dimefluron and its derivatives: see publications [27, 31, 32, 34, 39, 40, 52, 61, 71].
  • 1993-up to now: development and validation of new, original bioanalytical methods for the  bioequivalence studies, i.e. comparison of pharmacokinetics after oral administration of two dosage forms (reference vs. test). containing the identical amounts of medicament, but the dosage forms could differ in excipients used in specific dosage form.  
    Some of the mentioned methods were published in analytical journals (ambroxol [13], tramadol [16,33], fenofibrate [21], ursodeoxycholic acid [30], nabumetone [37, 38, 65, 66, 68], ciprofloxacin [41], 5-aminosalicylic acid [43], tiapride [62]. These papers had in most cases satisfactory citation response, two publications related to tramadol [16,33] were cited 97 times (calculated to January 2016). Some of the publications were awarded by the Czech Society for Experimental and Clinical Pharmacology and Toxicology of the Czech Medical Association of J.E. Purkyně (see next pages).    
    The developed and validated bioanalytical HPLC methods were employed not only in bioequivalence and pharmacokinetic studies for PRO.MED.CS Praha a.s., but also in the basic research of the fate of drugs in organism  (ADME studies of drugs with extensive metabolism: 5-ASA, fenofibrate, flubendazole, nabumetone, tiapride, tramadol).

 

Education and professional carrier

  • 1975-1980 : study at the Charles University, Faculty of Pharmacy in Hradec Králové, technological pharmacy branch (oriented for the research and pharmaceutical industry).

The employments chronologically:

  • 1980-1981: compulsory military service
  • 1981-1982: technical worker and later research worker at the Department of Biochemistry, Faculty of Pharmacy, Charles University, Hradec Králové (at prof. I. M. Hais and prof. E. Kvasničková).
  • 1982 - obtained degree PharmDr. based on the result of rigorous examination from pharmaceutical chemistry (Prof. Karel Palát and Assoc. Prof. Bohuslav Melichar)
  • 1983-1988 - postgradual study (assistantship) in the branch of pharmaceutical chemistry. In 1988 - written and defended CSc. thesis "Metabolites of potential  antineoplastic benfluron - synthesis, identification and determination". (CSc. = equivalent of Ph.D.)
  • January 1990 – relocation of my family from Kolín to Hradec Králové, into a new flat obtained from the Academy of Sciences
  • 1990–June 2013: Research worker at the Institute of Experimental Biopharmaceutics of the Czechoslovak Academy of Sciences in Hradec Králové (prof. Květina). After the splitting of Czechoslovakia to Czech and Slovak republic (1993), the Institute was renamed to Institute of Experimental Biopharmaceutics, Joint Research Centre of  Academy of Sciences of the Czech Republic and PRO.MED.CS. a.s. Praha).
  • Part-time job at the Department of Biochemical Sciences of Faculty of Pharmacy: common research projects and also the teaching of biochemistry (selected lectures).
  • 1995–2002 – part-time job at Faculty of Pharmacy offered by Prof. Karel Waisser (practice, seminars, selected lectures from organic chemistry at the Department of Inorganic and Organic Chemistry.
  • May 1998 Habilitation work “Synthetic, xenobiochemical and bioanalytical methodical approaches in preclinical and clinical research of selected drugs” was defended at Faculty of Pharmacy in Hradec Králové. Appointment by Associate Professor of Bioorganic Chemistry from 22nd June 1998.
  • From 1st January 2001 - head of analytical and bioanalytical laboratories of Institute of Experimental Biopharmaceutics, Joint Research Centre of  Academy of Sciences of the Czech Republic and PRO.MED.CS. a.s. Praha in Hradci Králové.
  • 2002-June 2013 – part-time job at the Department of Pharmaceutical Chemistry and Drug Control offered by Prof. J. Klimeš. Full teaching of the subjects „Analysis of  exogenous compounds (xenobiotics) in biomatrices“ and „Good Laboratory Practice“ for the students of the Health Care Bioanalytics (both present and distant form of the study).
  • From July 2013: full-time job at the Department of Pharmaceutical Chemistry and Drug Control offered by Prof. J. Klimeš, the employment at the Institute of Experimental Biopharmaceutics was terminated. Full return to Faculty of Pharmacy.
  • 10th June 2014 – professorial lecture „Study of the fate of drugs in the body from the perspective of bioanalyst“ at the Scientific Council of Faculty of Pharmacy; 30th October 2014 - short communication to the members of  Scientific Council of Charles University.
  • 1st May 2015  Professor Appointment

Teaching Experience

Lectures, practice, testing:

  • 1982-1989 – management practices of general biochemistry
  • 1990–1995 selected lectures of general biochemistry
  • 1996 – 2002 complete teaching of organic chemistry (management practices, seminars, testing)
  • 2002 – up to now  : complete training courses "Analysis of exogenous compounds in biological material" and "Good Laboratory Practice” (both present and combined form of the study in Healthcare Bioanalytics.
  • Selected chapters from „Therapeutic Drug Monitoring“ and „Pharmaceutical Analysis I+II“

Mentor of nineteen diploma or bachelor works.

Supervisor of Ph.D. theses (3 of them finished and successfully defended):

  • PharmDr. Přemysl Císař, Ph.D.: „Bioanalytické metodické přístupy při sledování osudu nového potenciálního léčiva v organismu pomocí HPLC-PDA-MS-MS“ (Bioanalytical methodical approaches to the monitoring of the fate of new potential drug in organism using  HPLC-PDA-MS-MS). Defended: 11th February 2008
  • PharmDr. Jana Maláková-Kopecká, Ph.D.: „Kvantitativní analýza léčiv v biologických materiálech separačními metodami“(Quantitative analysis of drugs in biomatrices by separation methods). Defended: 8th March 2012
  • Ing. Miloš Hroch, Ph.D.: „Vývoj a validace HPLC metod pro stanovení endogenních a exogenních látek pro transportní a metabolické studie“(Development and validation of HPLC method for the determination of endogenous and exogenous compounds for transport and metabolic studies). Defended: 8th March 2012
  • Next postgradual student: RNDr Zuzana Vybíralová

Grands received

(co-investigator responsible for the synthesis of potential drugs and their metabolites. HPLC analysis of compounds of interest in various biomatrices)

  • 1993-1995: Investigator prof. E. Kvasničková; GAČR 303/93/2021 „Possible improvements in chemical structure of cancerostatic based on pharmacokinetics and xenobiochemical findings“
  • 1993-1995: Investigator prof. P. Anzenbacher; GA AV ČR, grant No. A784103 „Biotransforma-tion  of xenobiotics by cytochromes P450 and by peroxidases“.
  • 1997-1999: Investigator prof. Vl.Geršl; IGA MZ ČR grant 4212-3/97 „New cytostatics – the issue of their cardiotoxicity and potential cardiprotective efficacy“

Garant and  co-investigator of two grants of Mgr. Přemysl Císař:

  • 2005-2006 GAUK 294/2005/B-CH „Analysis of potential antineoplastic dimefluron“
  • 2005-2006 FRVŠ 76/2005 „Use of different types of HPLC columns in bioanalysis of drugs“

Co-investigator of the grants of prof. Lenka Skálová :

  • 1998-2000 GAUK 247/1998/C; 2000-2002 GAČR 524/00/0514; 2003-2005 GAČR  524/03/1361;  2006-2008 GAČR     524/06/1345.

Co-investigator of the grants of assoc. prof. Barbora Szotáková :

  • 2005-2006  GAUK   108/2005/C; 2007-2009  GAČR 524/07/0611.    
  • 2005-2011 Research Project MSM 0021620822:„Research of new drug structures“

Cooperations

  • Prof. Ing. Michal Holčapek,Ph.D.; Assoc. Prof. Ing. Lenka Česlová-Kolářová, Ph.D.; Ing. Miroslav Lisa Ph.D. and Ing. Robert Jirásko, Ph.D. (Department of Analytical Chemistry, Faculty of Chemical Technology, University of Pardubice) in the field of LC-MS/MS identification of phase I and phase II drug metabolites. Some new, as yet undescribed combined conjugates were detected in some pig biomatrices. These conjugates of the type „nabumetone phaseI metabolite-glucuronide-glycine“ have a peptide bond between –COOH of glucuronide and H2N- of glycine [68].
     
  • Prof. RNDr. Milan Pour, Ph.D. and Assoc. Prof. PharmDr. Jiří Kuneš, Ph.D. (Department of Inorganic and Organic Chemistry, Faculty of Pharmacy in Hradec Králové, Charles University in Prague). Biotransformation study of potential antifungal agent 3-(4-bromophenyl)-5-acetoxymethyl-2,5-dihydrofuran-2-one in mice led to the identification of its principal phase I and phase II metabolites and revealed  the responsibility of one of the metabolites („olefin-18“) for its antifungal activity [47, 59]. NMR, IR and MS spectra of all synthesized standards for our studies were measured and interpreted at the Department of Inorganic and Organic Chemistry.
     
  • Prof. RNDr. Lenka Skálová, Ph.D. and Assoc. Prof. Ing. Barbora Szotáková, Ph.D. (Department of Biochemical Sciences, Faculty of Pharmacy in Hradec Králové, Charles University in Prague).
    ADME studies of benzo[c]fluorene antineoplastics (benfluron, dimefluron), ADME studies of benzimidazole anthelmintics. New achiral and chiral HPLC methods for the determination of flubendazole and its metabolites for „in vitro“ [46] and „in vivo“ [57] studies were developed and validated. The stereospecificity in metabolic reduction of flubendazole in various species (parasitic lancet fluke Dicrocoelium dendriticum, hosts pig, pheasant and ovine) was discovered.                    (+)-Enantiomer of reduced flubendazole was found to be the dominant flubendazole metabolite. 
     
  • Prof. Dr. Thomas Jira and Chamseddin Chamseddin, Ph.D. (Pharmazeutische/Medizinische Chemie, Institut für Pharmazie, Friedrich-Ludwig-Jahn-Str.17, 17489 Greifswald, Germany); Assoc. Prof. RNDr. Petr Bednář, Ph.D. (Department of Analytical Chemistry, Faculty of Science, Palacky University, Olomouc).  Identification of drug/metabolite enantiomers using HPLC equipped with polarimetric (circular dichroism) detector.
     
  • Prof. Ing. Vladimír Wsól, Ph.D. and Dr. Lucie Zemanová -Škarydová:
    Study of the stereospecificity of enzymes involved in carbonyl reduction of non-steroidal anti-inflammatory prodrug nabumetone [65,68]. Preferential carbonyl reduction of nabumetone to (+)-enantiomer of reduced nabumetone was proved.
     
  • Milan Nobilis dealt with various aspects of nabumetone biotransformation [37, 38, 65, 66, 68] in the cooperation with above-mentioned colleagues,:
    Chemical standards of all up to date known nabumetone phase I metabolites were synthesized and a LLE(SPE)-HPLC-PDA-FL-MS/MS method enabling the separation and determination of nabumetone, its five metabolites and naproxen (internal standard) within twelve-minute analysis was developed and validated. Some new, as yet undescribed combined conjugates were detected in some pig biomatrices. These conjugates of the type „nabumetone phaseI metabolite-glucuronide-glycine“ have a peptide bond between –COOH of glucuronide and H2N- of glycine [68]. Finally, a chiral HPLC method enabling the separation of the enantiomers of  reduced nabumetone and O-desmethyl-reduced nabumetone [68]. This method was used in the studies described in previous subsection (prof. Wsól and Dr. Škaridová [65].
    Within the above-described achiral and chiral HPLC analyses of the extracts from rat and human liver microsomes incubated with nabumetone, a missing link in nabumetone → 6-methoxy-2-naphthylacetic acid (6-MNA) biotransformation was revealed. This new, as yet undescribed nabumetone phase I metabolite was identified as 3-hydroxy nabumetone, which is a metabolic intermediate between prodrug nabumetone and pharmacodynamically active metabolite, 6-methoxy-2-naphthylacetic acid (6-MNA) [66]. Also, some new products of carbonyl reduction and O-desmethylation of 3-hydroxy nabumetone and 3-hydroxy nabumetone glucuronide were described in microsomes [66].

Publications

[1]    Kvasničková E., Nobilis M., Hais I.M.: Chromatographic characterization of in vitro metabolites of 5-(2-dimethylaminoethoxy)-7-oxo-7H-benzo[c]fluorene. J.Chromatogr. 295 (1984) 201-209

[2]    Nobilis M., Kvasničková E., Šroler A., Hais I.M.: Liquid chromatographic methods in the elucidation of the structure and quantitation of the metabolites of  benflurone. In: Proc. New Advances in Liquid Chromatography, Acad. Kiadó, Budapest (1986) 25-28

[3]    Nobilis M., Vančurová I., Hais I.M., Kvasničková E., Křepelka J.: Příprava některých 5-substituovaných derivátů 7-oxo-7H-benzo[c]fluorenu a 7-hydroxy-7H-benzo[c]fluorenu - potenciálních metabolitů benfluronu. Českoslov. Farm. 35 (1986) 68-73

[4]    Kvasničková E., Nobilis M., Šroler A., Báčová E., Hais I.M.: Chromatographic and enzymatic evidence for the structure of an oxygenated and reduced metabolite of "Benflurone". J.Chromatogr. 387 (1987) 559-561

[5]    Lyčka A., Jirman J., Nobilis M., Kvasničková E., Hais I.M.: Two-dimensional 1H and 13C -NMR spectra of 5-(2-dimethylaminoethoxy)-7-oxo-7H-benzo[c]fluorene, its precursor and metabolite. Magnetic Resonance in Chemistry 25 (1987) 1054-1057

[6]    Kvasničková E., Šroler A., Nobilis M.: Izolované hepatocyty jako model pro studium metabolismu léčiv. Českoslov. Farm. 37 (1988) 272

[7]    Nobilis M., Hais I.M.: Direct absorptiometric quantitation in high performance liquid chromatography in the absence of concomitant reference standards. J. Chromatogr. 434 (1988) 363-375

[8]    Hais I.M., Nobilis M., Kvasničková E.: Evaluation of Photometric HPLC Data for the Estimation of Benflurone Metabolites in Biological Materials with and without Concomitant Use of Standard. J. Chromatogr. 500 (1990) 643-653

[9]    Kvasničková E., Hais I.M. and Nobilis M.: Relationship between NADH and NADPH as cosubstrates of N-oxidation of the cytostatic benflurone by FMO. Progress in Pharmacology and Clinical Pharmacology. Vol. 8/3, Gustav Fisher Verlag, Stuttgard, New York (1991) 137-146.

[10]   Miko M., Křepelka J. Nobilis M.: 9-Hydroxybenfluron: A new in vivo Metabolite of Benfluron. In.: Recent Advances in Chemotherapy (Edit. Dieter Adam), Futuramed Publishers, Munich, Germany (1992) 2620-2621

[11]   Miko M., Křepelka J. and Nobilis M.: 9-hydroxy-benfluron: Effect on energy-yielding processes in Ehrlich ascites and P388 murine leukemia cells. Anti-cancer Drugs Vol.3 No.1 (1992) 63-68

[12]   Nobilis M., Kvasničková E., Šroler A. and Hais I.M.: Elimination of benfluron and its metabolites in the faeces and urine of rats. Drug Metabolism and Drug Interactions Vol.9, No.3-4 (1991) 225-240 (released in September 1992)

[13]   Nobilis M., Pastera J., Macek K., Svoboda D. and Květina J.: HPLC determination of ambroxol in human plasma. J. Chromatogr.-Biomedical Applications 581 (No.2) (1992) 251-255

[14]   Kvasničková E., Nobilis M., Šroler A., Wsól V. and Hais I.M.: In vitro and in vivo biotransformation of the potential cytostatic drug benfluron. In:"In vitro and Ex vivo Test Systems to Rationalize Drug Design and Delivery", Crommelin D., Couvreur P., Duchéne D. (eds.), Editions de Santé, Paris (1994) 331-335

[15]   Nobilis M., Anzenbacher P., Pastera J., Svoboda Z., Hrubý K., Květina J., Ubik K., Trejtnar F.: „Study of the biotransformation of a potential benzo[c]fluorene antineoplastic using high-performance liquid chromatography with high-speed scanning ultraviolet detection“, J. Chromatography B,  681 (1996) 143-151

[16]   Nobilis M., Pastera J., Anzenbacher P., Svoboda D., Kopecký J., Perlík F.: „High-performance liquid chromatographic determination of tramadol in human plasma“,  J. Chromatography B, 681 (1996) 177-183

[17]   Skálová L., Nobilis M., Szotáková B., Trejtnar F. and Kvasničková E.: „In vivo and in vitro biotransformation of benfluron N-oxide, a biologically active metabolite of a potential cytostatic“, Exp. Toxic. Pathol. Supplement II, 48 (1996) 366-370

[18]   Kvasničková E., Šroler A., Nobilis M., Wsól V., Machala M., Vortel V., Brejcha A. a Hais I.M.: „Activities of some biotransformation enzymes in the course of experimental pancreatitis. Chem. Listy 90(No.9) (1996) 743-744

[19]   Hrubý K., Anzenbacherová E., Anzenbacher P. and Nobilis M.: “Potential cancerostatic benfluron is metabolized by peroxidase : In vitro biotransformation of benfluron by horseradish peroxidase”, Gen. Physiol. Biophys. 16 (1997) 321-327

[20]   Nobilis M., Gregor J. and Waisser K.: „Examination of hydrophobic properties of antimycobacterially active 3-phenyl-2H-1,3-benzoxazine-2,4(3H)-diones, 3-phenyl-4-thioxo-2H-1,3-benzoxazine-2(3H)-ones and 3-phenyl-2H-1,3-benzoxazine-2,4(3H)-dithiones.“ Folia Pharmaceutica Universitatis Carolinae, XXIII Suppl. (1998) 89-90

[21]   Nobilis M., Květina J., Anzenbacher P., Vontor T., Zadák Z., Bláha V., Vlček J., Brátová M., Solichová D. a Svoboda D.: „Distribution of fenofibric acid in lipoprotein fractions of patients (and minipigs)“,  European Journal of Drug Metabolism and Pharmacokinetics, Vol.23, No.2 (1998) 287-294.

[22]   Skálová L., Nobilis M., Szotáková B., Wsól Vl. and Kvasničková E.: “Induction and inhibition of cytochrome P450-catalysed reduction of biologically active benfluron      N-oxide”. Drug Metab. Drug Interact. Vol.14, No.4 (1998) 221-234

[23]   Skálová L., Nobilis M., Szotáková B., Wsól Vl. and Kvasničková E.: “Inter-species comparison of microsomal reductive transformation of biologically active benfluron       N-oxide”. Drug Metab. Drug Interact. Vol.14, No.4 (1998) 235-250

[24]   Geršl Vl., Mělka M., Vortel J., Kvasničková E., Nobilis M. Mazurová Y. Hrdina R.: „New cytostatic agents - problems of their cardiotoxicity and potential cardioprotective activity“, Acta Medica (Hradec Králové) Vol. 41, No.2 (1998) 47-48

[25]   Květina J., Svoboda Z., Nobilis M., Pastera J., Anzenbacher P.: „Experimental Goettingen Minipigs and beagle dog as two species used in bioequivalence studies for clinical pharmacology (5-aminosalicylic acid and atenolol as model drug).“ Gen. Physiol. Biophys. 18 (1999) 80-85

[26]   Svoboda Z., Nobilis M., Květina J., Lemr K.: „Study of the biotransformation of benfluron using the isolated rat liver.“ Acta Medica (Hradec Králové) 42 (1999) 73-78

[27]   Macháčková J., Geršl Vl., Hrdina R., Nobilis M., Palička Vl.: „The influence of dimethoxybenfluron on biochemical and haematological parameters in rabbit.“ Acta Medica (Hradec Králové) 42 (1999) 89-92

[28]   Skálová L., Nobilis M., Szotáková B., Wsól Vl., Kubíček Vl., Baliharová V., Kvasničková E.: „Effect of substituents on microsomal reduction of benzo[c]fluorene N-oxides.“ Chemico-Biological Interaction 126 (2000) 185-200. Elsevier Science, Ireland Ltd.

[29]   Hrubý K., Anzenbacherová E., Anzenbacher P., Nobilis M.: „Biotransformation of benfluron by rat hepatic cytochrome P450. Identification of individual CYP-enzymes involved in biotransformation of benfluron, prospective antineoplastic based on benzo[c]fluorene.“ Collect. Czech. Chem. Commun. Vol. 65 (2000) 1374-1386

[30]   Nobilis M., Pour M., Kuneš J., Kopecký J., Květina J., Svoboda Z., Sládková K. and Vortel J.: „High-performance liquid chromatographic determination of ursodeoxycholic acid after solid phase extraction of blood serum and detection-oriented derivatization“,    J. Pharm. Biomed. Anal. 24 (2001) 937-946

[31]   Macháčková J., Adamcová M., Mazurová Y., Hrdina R., Nobilis M.: „Evaluation of Cardiac Effects of the New Antineoplastic Drug – Dimethoxybenfluron – in the Rabbit“, Physiological Research 50 (2001) 491-499

[32]   Adamcová M., Macháčková J., Geršl Vl., Hrdina R., Mazurová Y., Mělka M., Nobilis M.: „Plasma levels of cardiac troponin T following combination of daunorubicin with new antineoplastic drugs (Oracin and  Dimefluron) in rabbits“,  Acta Medica (Hradec Králové) Supp. (2001) 3-8

[33]   Nobilis M., Kopecký J., Květina J., Chládek J., Svoboda Z., Voříšek V., Perlik F., Pour M., Kuneš J.,: „High-performance liquid-chromatographic determination of tramadol and its O-desmethylated metabolite in blood plasma. Application to a bioequivalence study in humans.”, J. Chromatogr. A, 949 (1-2) (2002) 11-22 (ISSN 0021-9673)

[34]   Skálová L., Nobilis M., Szotaková B., Kondrová E., Šavlík M., Wsól V., Pichard-Garcia L., Maser E.: „Carbonyl reduction of the potential cytostatic drugs benfluron and 3,9-dimethoxybenfluron in human in vitroBiochem Pharmacol 64 (2002) 297-305

[35]   Pávek P., Štaud F., Fendrich Z., Sklenářová H., Libra A., Novotná M., Kopecký M., Nobilis M., Semecký V.: „Examination of the Functional Activity of P-glycoprotein in the Rat Placental Barrier Using Rhodamine 123.“ The Journal of Pharmacology and Experimental Therapeutics Vol.305, No.3 (2003) 1239-1250

[36]   Eva Anzenbacherová, Karolina Filipová, Milan Nobilis, Pavel Anzenbacher :"Selective determination of famotidine in human plasma by high performance liquid chromatography in alkaline media with solid phase extraction", Journal of Separation Science 26 (No.8) (2003) 722-726

[37]   Nobilis M., Kopecký J., Květina J., Svoboda Z., Pour M., Kuneš J., Holčapek M., Kolářová L.: „Comparative biotransformation and disposition studies of  nabumetone in humans and minipigs using high-performance liquid chromatography with ultraviolet and mass-spectrometric detection“. J. Pharm. Biomed. Anal. Vol. 32/4-5, 641-656 (2003)

[38]   Nobilis M., Holčapek M., Kolářová L., Kopecký J., Kuneš M., Svoboda Z., Květina J.: "Identification and determination of phase II nabumetone metabolites by high-performance liquid chromatography with photodiode-array and mass spectrometric detection." J. Chromatogr. A 1031/1-2  (2004) 229-236  (ISSN 0021-9673)

[39]   Císař P., Nobilis M., Vybíralová Z., Holčapek M., Kolářová L., Pour M., Kuneš J., Klimeš J.: „Disposition study of a new potential antineoplastic agent dimefluron in rats using high-performance liquid chromatography with ultraviolet and mass spectrometric detection.“ J. Pharm. Biomed. Anal. Vol. 37/5 (2005) 1059-1071.

[40]   Kučera R., Nobilis M., Skálová L., Szotáková B., Císař P., Jira Th., Klimeš J., Wsól V.: „Use of chiral reversed-phase liquid chromatography for the evaluation of stereoselectivity in the carbonyl reduction of potential benzo[c]fluorene antineoplastics benfluron and dimefluron in various species.“ J. Pharm. Biomed. Anal. Vol. 37/5 (2005) 1049-1057.

[41]   Vybíralová Z., Nobilis M., Zoulová J., Květina J., Petr P.: „High-performance liquid chromatographic determination of ciprofloxacin in plasma samples.“ J. Pharm. Biomed. Anal. Vol. 37/5 (2005) 851-858.

[42]   Slanař O., Nobilis M., Květina J., Idle J.R, Perlík F.: „CYP2D6 polymorphism, tramadol pharmacokinetics and pupillary response“, Eur. J. Clin. Pharmacol. 62 (2006) 75-76

[43]   Nobilis M., Vybíralová Z., Sládková K., Lísa M., Holčapek M., Květina J.: „High-performance liquid-chromatographic determination of 5-aminosalicylic acid and its metabolites in blood plasma.“ J. Chromatogr. A 1119 (2006) 299-308

[44]   Slanař O., Nobilis M., Květina J., Olga Matoušková O., Jeffrey R. Idle J.R.., Perlík F.: „Pharmacokinetics of tramadol is affected by MDR1 polymorphism C3435T“, European Journal of Clinical Pharmacology 63 (2007) 419-421

[45]   Slanař O., Nobilis M., Květina J., Mikoviny R., Zima T.,  Idle J.R.., Perlík F.: „Miotic Action of tramadol is determined by CYP2D6 genotype, Physiol. Res. 56 (2007) 129-136

[46]   Nobilis M., Jira T., Lísa M., Holčapek M., Szotáková B., Lamka J., Skálová L.: „Achiral and chiral high-performance liquid chromatographic determination of flubendazole and its metabolites in biomatrices using UV photodiode-array and mass spectrometric detection.“ Journal of Chromatography A 1149 (2007) 112-120

[47]   Nobilis M., Pour M., Pavlík J., Šenel P., Kuneš J., Vopršalová M., Kolářová L., Holčapek M.:„Metabolic profiling of a potential antifungal drug, 3-(4-bromophenyl)-5-acetoxymethyl-2,5-dihydrofuran-2-one, in mouse urine using high-performance liquid chromatography with photodiode-array and mass spectrometric detection.“ Journal of Chromatography B  853 (2007) 10-19

[48]   Maláková J., Nobilis M., Svoboda Z., Lísa M., Holčapek M., Květina J., Klimeš J., Palička V.: „High-performance liquid chromatographic method with UV-photodiode array, fluorescence and mass spectrometric detection for simultaneous determination of galantamine and its phase I metabolites in biological samples.“ Journal of Chromatography B   853 (2007) 265-274

[49]   Kuneš M., Květina J., Kubant P., Nobilis M., Šmídová I., Herout V., Svoboda Z.: „The rat small intestine in situ perfusion technique as a tool for bioequivalence estimation of suspension drug forms.“ Biomedical Papers 151, Supp.1 (2007) 47-50 (Formerly Acta Universitatis Palackianae Olomucensis Facultatis Medicae, ISSN 1213-8118)

[50]   Matal J., Anzenbacherová E., Veinlichová A., Nobilis M., Anzenbacher P.: „Interaction of nabumetone with cytochromes P450 in vitro: comparison of the man and pig.“ Biomedical Papers 151, Supp.1 (2007) 53-54 (Formerly Acta Universitatis Palackianae Olomucensis Facultatis Medicae, ISSN 1213-8118)

[51]   Cvilink V., Kubíček V., Nobilis M., Křížová V., Skotáková B., Lamka J., Várady M., Kuběnová M., Novotná R., Gavelová M., Skálová L.: „Biotransformation of flubendazole and selected model xenobiotics in Haemonchus contortus“. Veterinary parasitology 151 (2008) 242-248

[52]   Holčapek M., Kolářová L., Nobilis M.: „High-performance liquid chromatography – tandem mass spectrometry in the identification and determination of phase I and phase II drug metabolites.“ Anal. Bioanal. Chem. 391 (2008) 59-78

[53]   Křížová V., Lamka J., Szotáková B., Vokřál I., Srpová V., Urbánková M., Kubíček V., Nobilis M., Skálová L.: „Dicrocoeliosis of  old mouflon ewes – effect on biotransformation enzymes and metabolism of anthelmintics in vitro.“ The Open Veterinary Science Journal 2 (2008) 23-31

[54]   Szotáková B., Nobilis M., Lamka J., Křížová V., Šavlík M., Skálová L.: „Modulation of porcine (Sus scrofa domestica) and pheasant (Phasianus colchicus) carbonyl reducing enzymes by althelmintic therapy with flubendazole“. Drug Metabolism Letters 2 (2008) 29-34

[55]   Kubíček V., Soukupová M., Nobilis M., Křížová V., Szotáková B., Skálová L.: „LC with fluorimetric detection for sensitive analysis of reduced flubendazole in biological samples“. Chromatographia 68 (2008) 865-867

[56]   Vybíralová Z., Nobilis M., Kholová D., Svoboda Z.: „Usage LC-MS Technique for determination of enalapril and its metabolite enalaprilat in pharmacokinetic studies.“ Prague Medical Report Vol.109 (2008) Suppl., pp.129-131

[57]   Nobilis M., Vybíralová Z., Křížová V., Kubíček V., Soukupová M., Lamka J., Szotáková B., Skálová L.: „Sensitive chiral high-performance liquid chromatographic determination of anthelmintic flubendazole and its phase I metabolites in blood plasma using UV photodiode-array and fluorescence detection. Application to pharmacokinetic studies in sheep.“ J. Chromatogr. B 876 (2008) 89-96

[58]   Křížová V., Nobilis M., Prušková L., Chládek J., Szotáková B., Cvilink V., Skálová L., Lamka J.: „Pharmacokinetics of flubendazole and its metabolites in lambs and adult sheep (Ovis aries)“ Journal of Veterinary Pharmacology and Therapeutics 32(6) (2009) 606-612; DOI 10.1111/j.1365-2885.2009.01082.x

[59]   Šenel P., Tichotová L., Votruba I., Buchta Vl., Špulák M., Kuneš J., Nobilis M., Krenk O., Pour M.: „Antifungal 3,5-disubstituted furanones: From 5-acyloxymethyl to 5-alkylidene derivatives“, Bioorganic & Medicinal Chemistry 18 (2010) 1988-2000, IF(2009) = 2.822

[60]   Vokřál I., Křížová  V., Lamka J., Kubíček V., Szotáková B, Várady M., Nobilis M., Skálová L.: "Effect of Flubendazole on Biotransformation Enzymes Activities in Haemonchus contortus". The Open Parasitology Journal,  4 (2010) 24-28 (new journal without IF yet)

[61]   Jirásko R., Holčapek M., Nobilis M.: „Identification of Phase I and Phase II metabolites of benfluron and dimefluron in rat urine using high-performance liquid chromatography - tandem mass spectrometryRapid Commun. Mass Spectrom. 25 (2011) 2153–2162.
DOI: 10.1002/rcm.5097

[62]   Nobilis M., Vybíralová Z., Szotáková B., Sládková K., Kuneš M., Svoboda Z.: „High-performance liquid chromatographic determination of tiapride and its phase I metabolite in blood plasma using UV photodiode-array and fluorescence detection in tandem. Application to pharmacokinetic studies“ J. Chromatogr.B  879 (2011) 3845-3852.
DOI: 10.1016/j.jchromb.2011.10.032

[63]   Stariat J., Šesták V., Vávrová K., Nobilis M., Kollárová Z., Klimeš J., Kalinowsky D.S., Richardson R..D.: Kovaříková P., „LC-MS/MS identification of the principal in vitro and in vivo phase I metabolites of the novel thiosemicarbazone anti-cancer drug, Bp4eT“. Anal Bioanal Chem 403 (2012) 309-321.  DOI 10.1007/s00216-012-5766-4

[64]   Hroch M., Havlínová Z., Nobilis M. Chládek J.: “HPLC determination of arginases inhibitor N-(w)-hydroxy-nor-L-arginine using core-shell particle column and LC-MS/MS identification of principal metabolite in rat plasma” J. Chromatogr.B  880 (2012) 90-99.
DOI: 10.1016/j.jchromb.2011.11.022

[65]   Škarydová L., Nobilis M., Wsól V.: „Role of carbonyl reducing enzymes in the phase I biotransformation of the NSAID nabumetone in vitro.“ Xenobiotica  43 (2013) 346-354

[66]   Nobilis M., Mikušek J., Szotáková B., Jirásko R., Holčapek M., Chamseddin C., Jira T., Kučera R., Kuneš J., Pour M., „Analytical power of LLE-HPLC-PDA-MS/MS in drug metabolism studies: Identification of new nabumetone metabolites.” J. Pharm. Biomed. Anal. 80 (2013) 164-172.

[67]   Smetanová L., Štětinová V., Kholová D., Kuneš M., Nobilis M., Svoboda Z., Květina J.: „Transintestinal transport mechanisms of 5-aminosalicylic acid (in situ rat intestine perfusion, Caco-2 cells) and Biopharmaceutics Classification System“. Gen. Physiol. Biophys.32 (2013) 361–369.

[68]   Česlová L., Holčapek M., Nobilis M.: „Identification of combined conjugation of nabumetone phase I metabolites with glucuronic acid and glycine in minipig biotransformation using coupling high-performance liquid chromatography with electrospray ionization mass spectrometryJ. Pharm. Biomed. Anal. 88 (2014) 221-224.

[69]   Stariat J., Suprunová V., Roh J., Šesták V., Eisner T., Filipský T., Mladěnka P., Nobilis M., Šimůnek T., Klimeš J., Kalinowski D.S., Richardson D.R., Kovaříková P.: „Simultaneous determination of the novel thiosemicarbazone anti-cancer agent, Bp4eT, and its main phase I metabolites in plasma: Application to a pilot pharmacokinetic study in rats“. Biomed Chromatogr.28/5 (2014) 621-629. Doi: 10.1002/bmc.3080.

[70]   Kuneš M., Květina J., Tachecí I., Kopáčová M., Bureš J., Nobilis M., Krejcar O., Kuča K.: „Imaging and Evaluating Method as Part of Endoscopical Diagnostic Approaches“ Edited by: Nguyen N.T., Attachoo B., Trawinski B., Somboonviwat K. Intelligent Information and Database Systems, PT II. Book Series: Lecture Notes in Artificial Inteligence; Volume: 8398 (2014) pp. 605-614 (ISSN: 0302-9743), IF2005= 0.302.

[71]   Suchá L., Kolenič M., Kratochvíl J., Pour M., Nobilis M., Čermáková E., Řezáčová M., Tomšík P.: „Novel Derivatives of Benfluron and Dimefluron: Synthesis and Anticancer Activity“ Letters in Drug Design & Discovery Vol.12, Issue10 (2015) 787-801. IF2014= 0.770.

Patents:

1.       Vančurová I., Mělka M., Křepelka J., Nobilis M., Kvasničková E., Beneš M., Hais I.M., Skládaná M., Kejhová I., Pokorná S., Reichlová R.: N-Oxides of 5-(dialkyl-amino)alkoxy-7-oxo-7H-benzo[c]fluorene derivatives and the method of their production. Patent No. 239413, Bureau for inventions and discoveries (PV 625-84), Prague, 1988.

2.       Nobilis M., Vančurová I., Křepelka J., Mělka M., Lyčka A., Jirman J., Kvasničková E., Hais I.M., Miko M.: Positional isomers of x-hydroxy-5-(2-dimethylaminoethoxy)-7-oxo-7H-benzo[c]fluorene. Patent No. 267519, Federal Bureau for inventions (PV 02914-88), Prague, 1990.

WOS

Technical text:

1.       Drašar P. and Nobilis M.: Integrator SP4400 Chromjet (User Handbook). pds and  SciTech, Praha (Autumn 1992)

Qualification works:

  1. Nobilis Milan: „Dělení Pelentanu a jeho metabolitů metodou vysokotlaké kapalinové chromatografie“ („Separation of pelentan and its metabolites using HPLC“), Diploma Thesis. Charles University in Prague, Faculty of Pharmacy in Hradec Králové 1980, 89 pages.
  2. Nobilis Milan: „Analýza léčiv a jejich metabolitů v biologickém materiálu kapalinovou chromatografií“ („Analysis of drugs and their metabolites in biological materiál“). An overview to PhD., Charles University, Faculty of Pharmacy in Hradec Králové, April 1986, 47 pages
  3. Nobilis Milan: „Metabolity potenciálního antineoplastika benfluronu – syntéza, identifikace a stanovení“ (Metabolites of potential antineoplastic benfluron – synthesis, identification and determination), Ph.D. thesis, Charles University, Faculty of Pharmacy in Hradec Králové 1988, 213 pages.  
  4. Nobilis Milan: „Syntetické, xenobiochemické a bioanalytické metodické přístupy v preklinickém a klinickém výzkumu vybraných léčiv“ (Synthetic, xenobiochemical and bioanalytical methodical approaches in preclinical and clinical research of selected drugs), Habilitation work, 178 pages, Hradec Králové, September 1996

Membership in  professional societies:

  • Member of Czech Pharmaceutical Society 1996 – up to now
  • 1997 – 2002: Scientific Secretary of Synthetic Section Committee of Czech Pharmaceutical Society, 2002 – up to now: member of Synthetic Section Committee of Czech Pharmaceutical Society
  • Member of Czech Medical Association of J. E. Purkyně and Pharmacological Society: 2001 – up to now
  • Committee Member of Czech Society for Experimental and Clinical Pharmacology and Toxicology of the Czech Medical Association of J. E. Purkyně: 2006 – up to now

Awards:

Six of our publications were awarded by Czech Society for Experimental and Clinical Pharmacology and Toxicology of the Czech Medical Association of J. E. Purkyně.

First prize in the competition for the best scientific work in category of CLINICAL PHARMACOLOGY (4 times in the years 2002, 2003, 2006, 2011):  

  • Nobilis M, Kopecký J, Květina J, Chládek J, Svoboda Z, Voříšek V, Perlík F, Pour M, Kuneš J: “High-performance liquid-chromatographic determination of tramadol and its O-desmethylated metabolite in blood plasma. Application to a bioequivalence study in humans.” J Chromatogr.A 949 (2002) 11-22.
  • Nobilis M., Kopecký J., Květina J., Svoboda Z., Pour M., Kuneš J., Holčapek M., Kolářová L.: “Comparative biotransformation and disposition studies of  nabumetone in humans and minipigs using high-performance liquid chromatography with ultraviolet and mass-spectrometric detection”. J. Pharm. Biomed. Anal. 32 (2003) 641-656.
  • Nobilis M., Vybíralová Z., Sládková K., Lísa M., Holčapek M., Květina J.: „High-performance liquid-chromatographic determination of 5-aminosalicylic acid and its metabolites in blood plasma.“ J. Chromatogr. A  1119 (2006) 299-308.
  • Nobilis M., Vybíralová Z., Szotáková B., Sládková K., Kuneš M., Svoboda Z.: „High-performance liquid-chromatographic determination of tiapride and its phase I metabolite in blood plasma using tandem UV photodiode-array and fluorescence detection.“ J. Chromatogr. B 879 (2011) 3845-3852.

First prize in the competition for the best scientific work in category of EXPERIMENTAL PHARMACOLOGY (2 times in the years 2003, 2007):

  • Pávek P, Štaud F, Fendrich Z, Sklenářová H, Libra A, Novotná M, Kopecký M, Nobilis M, Semecký V.: “Examination of the functional activity of P-glycoprotein in the rat placental barrier using rhodamine 123.” J. Pharmacol. Exp. Ther. 305 (2003) 1239-1250
  • Nobilis M., Jira T., Lísa M., Holčapek M., Szotáková B., Lamka J., Skálová L.: „Achiral and chiral high-performance liquid chromatographic determination of flubendazole and its metabolites in biomatrices using UV photodiode-array and mass spectrometric detection.“ Journal of Chromatography A 1149 (2007) 112-120.

List of most cited publications (WoS – January 2016) :

Holčapek M., Kolářová L., Nobilis M.: „High-performance liquid chromatography – tandem mass spectrometry in the identification and determination of phase I and phase II drug metabolites.“ Anal. Bioanal. Chem. 391 (2008) 59-78 - Cited 113 x

Nobilis M., Pastera J., Anzenbacher P., Svoboda D., Kopecký J., Perlík F.: „High-performance liquid chromatographic determination of tramadol in human plasma“,  J. Chromatogr. B, 681 (1996) 177-183 - Cited 52 x

Vybíralová Z., Nobilis M., Zoulová J., Květina J., Petr P.: „High-performance liquid chromatographic determination of ciprofloxacin in plasma samples.“ J. Pharm. Biomed. Anal. Vol. 37/5 (2005) 851-858 - Cited 47 x

Nobilis M., Kopecký J., Květina J., Chládek J., Svoboda Z., Voříšek V., Perlik F., Pour M., Kuneš J.,: „High-performance liquid-chromatographic determination of tramadol and its O-desmethylated metabolite in blood plasma. Application to a bioequivalence study in humans.”, J. Chromatogr. A, 949 (1-2) (2002) 11-22 - Cited 45 x

Nobilis M., Vybíralová Z., Sládková K., Lísa M., Holčapek M., Květina J.: „High-performance liquid-chromatographic determination of 5-aminosalicylic acid and its metabolites in blood plasma.“ J. Chromatogr. A  1119 (2006) 299-308. - Cited 39 x

Stay abroad, trainings:

  • 1989 – study visit at E.M.Arndt University  in Greifswald and Humboldt University in Berlin
  • Foreign Trainings:
    • 1990, 1993 - Training HPLC systems (Spectra Physics, Darmstadt).
    • 2007 – Training LC-MS systems Thermo Finnigan in London.

Active participation at 29 foreign symposia (1993-2012):

  • 5th European Congress on Biopharmacy and Pharmacokinetics, Brussel, Belgium, April 20-22, 1993
  • 14th European Workshop on Drug Metabolism, Paris, July 3-8, 1994
  • 2nd European Congress on Specificity and Variability in Drug Metabolism (European Concerned Action COST  B1), May 10-12, 1995, Besançon, France
  • 6th European Congress of Biopharmacy and Pharmacokinetics in Athens, Greece, April 22-24, 1996
  • 15th European Workshop on Drug Metabolism, Jena, September 9-13, 1996
  • 6th European Congress of International Society for the Study of Xenobiotics, Göteborg (Sweden), June 30 - July 3, 1997
  • 16th European Workshop on Drug Metabolism, June 21-26, 1998, Copenhagen, Denmark.
  • 13th International Congress of Pharmacology, 26-31 July 1998, München, Germany.
  • 5th Congress of the European Federation of Pharmaceutical Sciences (EUFEPS), April 25-30, 1999 Jerusalem (Israel).
  • 7th European ISSX Meeting, Budapest (Hungary), August 22-26, 1999
  • 11th International Symposium on Pharmaceutical and Biomedical Analysis (PBA 2000), May 14-18, 2000, Basel, Switzerland
  • 25th International Symposium on High Performance Liquid Phase Separations and Related Techniques (HPLC 2001), 17.-22. června 2001, Maastricht,  Netherlands.
  • 6th International ISSX Meeting, October 7-11, 2001, Munich.
  • Drug Analysis 2002, Brugges, Belgium, 21-25 April 2002,
  • 27th Symposium on High Performance Liquid Phase Separations and Related Techniques (HPLC 2003), 15-19 June 2003, Nice, France
  • 15th International Symposium on Pharmaceutical and Biomedical Analysis (PBA2004), May 2-6, 2004, Florence, Italy.
  • 25th International Symposium on Chromatography (ISC04), October 4-8, 2004, Paris, France
  • 29th International Symposium on High Performance Liquid Phase Separations and Related Techniques (HPLC 2005), June 26-30, 2005, Stockholm (Sweden).
  • 11th Meeting on Recent Developments in Pharmaceutical Analysis (RDPA 2005), September 25-28, 2005, Rimini (Italy).
  • 9th European ISSX (International Society for the study of xenobiotics) Meeting, Manchester, UK, June 4-7, 2006.
  • 26th International Symposium on Chromatography (ISC06), August 21-25, 2006, Copenhagen (Denmark)
  • 31st International Symposium on High Performance Liquid Phase Separations and Related Techniques (HPLC 2007), June 17-21, 2007, Ghent, Belgium. Ghent
  • 10th European Regional ISSX Meeting, May 18-21, 2008, Vienna, Austria
  • 27th International Symposium on Chromatography, September 21-25, 2008, University of Münster, Germany
  • 20th International Symposium on Pharmaceutical and Biomedical Analysis, March 1-4, 2009, Agra, India
  • 11th European Regional ISSX Meeting, May 17-20, 2009 Portugal, Lisabon
  • 9th International ISSX Meeting, Istanbul Turkey, September 4-8, 2010
  • 36th International Symposium on High-Performance Liquid Phase Separations and Related Techniques (HPLC2011), Budapest , Hungary, 19th-23rd June 2011.
  • 29 th International Symposium on Chromatography, Toruň, Poland, September 9–13, 2012

© 2017 Charles University, Faculty of Pharmacy in Hradec Králové, Akademika Heyrovského 1203, 500 05 Hradec Králové, Czech Republic

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