BIOMEDICAL RESEARCH CENTER (BRC) UNIVERSITY HOSPITAL HK

Kamil Kuca

BIOMEDICAL RESEARCH CENTER

BRC

UH

FVZ

UHK Faf

LFHK

University Hospital HK

Faculty of Military Health Sciences

Pharmaceutical faculty

Faculty of Medicine

Faculty of Science UHK Faculty of Informactics and

Managment UHK (Biomedical Informatics

Center)

BIOMEDICAL RESEARCH CENTER

Brief history: •April 2011 - Established To prevent outflow of scientists (3 full positions divided into 6 people)

•February 2012 - Laboratory facility – EU postdoc

program started (4 full positions gained)

•October 2013 - 30 empoyees (senior and junior

scientists, PhD students, lab. assistants; total 3.2 positions – full/part time)

- total 50-60 people (students and other helpers)

BIOMEDICAL RESEARCH CENTER

AIMS:

Support basic research in the Hospital (drug development, analysis, assessment etc.)

Service for the clinics and applied research (proteomics, pharmacokinetics etc.)

Education (pre and postgraduated students, interships, seminars)

BIOMEDICAL RESEARCH CENTER

Projects: Currently solving 11 project (including internal) 6 internal cooperations (within Hospital) 8 external cooperations (in CZ)

Funding: • Grant Agency of CZ, Grant agencies under Ministry of Health, Defence,

Education etc. • Private sector (organic synthesis, cytotoxicity, acute toxicity etc)

Incomes: Projects (10-20% for the hospital) Publications (depending on the IF) Services for fee

...what we get, we need to pay back

BIOMEDICAL RESEARCH CENTER

Cooperations:

Canada

USA

Brazil

Croatia Sweden France Germany Serbia Hungary

Korea

Singapore

India

UAE

UK Italy Slovakia

...30 to 40 IF papers per year

Představení proteomické skupiny Centra Biomedicínského Výzkumu

Chemical

synthesis

Pharmacokinetics

Proteomics

In vitro biologial

assessment

D

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D

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o

p

m

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t

BIOMEDICAL RESEARCH CENTER

In vivo assesment,

toxicity

DRUG DEVELOPMENT

Drug discovery

Preclinical Trials

Clinical trials

(I.-III. phase)

Practical use

(IV. phase)

12-15 years, aprox. 1 mld USD

BRC

Targeted synthesis Design and Synthesis

In-vitro evaluation

Toxicity

1st selection

Pharmacokinetics

In-vivo

?

2nd selection

„Drug candidates“- recommendation for preclinical trials, commercialization, ip protection

BRC - DRUG DESIGN

BRC - DRUG DESIGN

• targeted synthesis (in silico methods – molecular docking, QSAR) • modification of known drugs • isolation from the plants

Topics

AChE MODULATORS

Alzheimer´s Myasthenia

gravis Antidotes

ABAD inhibitors

Alzheimer´s

Detergents

Desinfection

Decon Anticancer

MOLECULAR DOCKING

• Enzyme known structure (enzym, receptor)- RTG; www.pbd.org • docking of drug candidates into the enzyme cavity

N

NH2

N

NH2

H3CO

Tacrine 7-MEOTA IC50: 0.5 ± 0.1 uM IC50: 10.5 ± 2 uM

CHEMICAL SYNTHESIS

Organophosporus prophylaxis

Myastenia gravis

Charged: Peripherally active AChE inhibitors

N (A) N 2 X

R

HON=HC

ANTIDOTES

Oxime moiety

CHEMICAL SYNTHESIS

Alzheimer‘s disease Homodimers

Detergents Heterodimers

ABAD

Uncharged AChEi

IN VITRO SCREENING

Alzheimer‘ disease

• Inhibition of AChE, Beta-secretase, Prolyl-oligopeptidase,

• Affinity to M a N rcp.

• Antioxidant activity

• Inhibition of Abeta formation (SAV Kosice)

• Inhibition of ABAD (St. Andrews, UK)

• Etc.

AChE reactivators

• Inhibition of AChE

• Reactivation screening (10-3 a 10-

5M) against nerve agents

• Kinetics

• Corellation with standards (obidoxime, HI-6 etc.)

IN VITRO SCREENING

PERIPHERAL AChE INHIBITORS

• ACHE inhibitors for:

• Myasthenia gravis

• Prophylaxis against Nerve Agents

• STAT3 inhibitors

• ATM inhibitors

• Cooperation Academy of Science

DETERGENTS

•Desinfection means -CMC->MIC na G+ a G- strains, fungi

•Decontamination means -CMC

•Stabilisation of gold nanoparticles

•hypertermal anticancer therapy

•preparation of SERS active substrates

•Novel topic

•Novel adjuvans for vaccines (TLR4 receptor antagonists)

ANTICANCER THERAPY

ADJUVANTS

CYTOTOXICITY

IC50=5,14µM

TOXICITY

PHARMACOKINETICS

Clinical proteomics Quantitative and qualitative description of the protein content in the samples using LCMS Explorative analysis •What protiens are in a sample?= identification •What is the difference in quantities of different proteins in a samples (treated vs non-treated group) = quantification

Targeted analysis •I know what I´m looking for and what is the level of this protein in different samples

BRC - PROTEOMICS

PROTEOMICS –projects

1)Proteomic analysis of amniotic fluid samples from preterm birth patients 2)Protein markers of inflammatory complications during preterm birth 3) Proteomic analysis of different kind of thrombi in case of heart attack 4) Proteomic analysis of the rat brains chronically exposed to low doses of pesticides (gulf-war syndrom) 5) Risk assassment based on the proteomic analysis for the pituitary adenome relapses (retrospective study) 6) Proteomic analysis of different kind of colorectal cancers • With/without biological treatment • Retrospective study of the relepses of the CoCa

Tambor, V. et al. J Proteomics 2012, 3, 857 Tambor, V. et al. PLoS ONE 2012, 7

MILITARY PRODUCTS

BEFORE INTOXICATION

AFTER INTOXICATION

Nerve Agent

Prophylaxis (= Pretreatment) -Pyridostigmine, Physostigmine

-Transant (HI-6)

-PANPAL, etc.

Decontamination -reactive or non-reactive

-personal or mass

Treatment -anticholinergic drugs

-acetylcholinesterase reactivators

-anticonvulsants

MILITARY RESEARCH FOCUS

K-OXIMES

Reactivators in progress – K027 – excellent reactivator for OP pesticide poisoning

– K203 – excellent reactivator for tabun poisoning

AUTOINJECTOR

Three-chambered autoinjector

1. chamber - oxime HI-6

2. chamber - atropine

3. chamber - diazepam

Autoinjector patented.

www.chemprotect.eu

BChE REACTIVATORS

• Potent and „broad-spectrum“ reactivators of human BChE inhibited

with paraoxon, dichlorvos, sarin, cyclosarin, tabun and VX

• If possible, aim to develop broad spectrum reactivators for both

AChE and BChE

• More than 100 different structures

CH

NOH

N+

R1

n

R1

=

R2

N+

R2

,

Pseudo-catalytic bioscavengers

SMALL MOLECULE PROPHYLACTICS

NUCLEOPHILIC DETERGENTS

Detergent and

Nucleophile in

one Molecule Non-functionalized

Detergents

Non-functionalized Detergents

+ Nucleophile

+

Three different types of systems:

1. 2. 3.

• Skin penetration studies • In vitro x In vivo • Franz-type diffusion cells • Pigs, rats

0

0,02

0,04

0,06

0,08

0,1

0,12

0 100 200 300 400 500 600

Time (min)

Am

ou

nt p

en

etr

ate

d (

%)

5 min

60 min

2 min

30 min

permeation

Paraoxon

Micelle Creation:

DECONTAMINATION MEANS

HVĚZDA (trans. STAR)

• Bi-component mixture

• Decontamination of warfare agents G, V, H, biological agens – e.g.

spores of B. anthracis (antrax)

• Suitable for skin and common surfaces

Active substances:

Quaternary ammonium salts (as surfactant),

hydrogen peroxide

Users:

• Fire Rescue Service of CR, Army of CR, NATO NRF 3

(2004), civil use – broad spectrum disinfection

www.fnhk.cz/cbv

29

Thank you for your attention