The IEPOR facilities for conducting preclinical studies

RE Kavetsky IEPOR of NAS of Ukraine is the academic research center with almost 60 years of experience in the field of experimental pathology, biotechnology, molecular and clinical oncology.

The research team of IEPOR consists of more than 120 scientists, including one Academician and one corresponding member of NAS of Ukraine, 22 doctors of sciences, 64 PhDs and 18 PhD students, who are carrying out the scientific research projects, according to the current activities of the institute.

Scientific and methodological basis for scientific research consists of:

  • Center of Expertise for determination of carcinogenic factors and molecular markers of tumor growth
  • Center of specific, toxicological, pharmacodynamic and pharmacokinetic studies for preclinical antitumor, antiviral, antiangiogenic agents, and immunomodulators and radioprotectors
  • Center for quality control of medicines
  • Center of collective use of scientific equipment «Molecular Oncology and Biotechnology»

The following Working Groups are part of the abovementioned Centers:

  • Research in cell culture
  • Preclinical studies of anticancer agents
  • Identification of molecular biologic markers of tumor growth
  • Determination of carcinogenic factors and their genotoxic and mutagenic effects; biotechnology

The centers are certified and have the all required permits (certificate SE «Ukrmetrteststandart» number PT-400/14 of 28.11.2014, valid till to 27/11/2018).


Cell Lines Bank from human and animal tissues

IEPOR has its own «Cell Lines Bank (BCL) from human and animal tissues» — the official collection of cellular materials aimed at for collecting, cryogenic storage and distribution of cell cultures and the cultures of transplanted tumors. Bank collection contains more than 30,000 samples of typical and original cell lines from normal and tumor tissues of humans and different animal species — rats, mice, hamsters, monkeys, pigs, dogs, bulls, sheep, bats, mink, fish, insects, etc. It is more than 200 cell lines and more than 30 strains of transplanted tumors.

The modern methods of research are widely used in BCL: cells’ cultivating and cloning, cell transduction by different genes, and the cytological, immunologic, virologic, biochemical, morphologic and cytogenetic research methods.

Bank staff has extensive experience in testing in vitro and in vivo factors with antitumor and antiviral activity of various nature (chemical, physical, biological).

The BCL has appropriate cellular model systems to determine in vitro the biological activity of various cytokines (interleukins, growth factors, tumor necrosis factor, interferon, etc).

An important part of the scientific work is devoted to the study of biological and morphological properties of cells and their imunofenotypical and cytogenetic features.


Vivarium — experimental biological clinic

The Institute has the experimental biological clinic (vivarium), which is intended for the maintenance of laboratory animals and carrying out experimental work with their use. In the vivarium, breeding of laboratory animals (rats and mice) is also carried out. Among them:

mice

  • Nonlinear (mongrel) S57VL6 line (black) — low-cancer line, insensitive to milk factor
  • Line BALB/c (Balbey) — low-cancer line, sensitive to milk factor (Bittner virus)
  • Line DBA/2 — high-cancer line
  • Line CBA that was obtained by the cross-breeding of mice line BALB/c with mice line DBA/2

rats

  • Nonlinear
  • Wistar line (Wistar) — originates from the colony of rats-albinos, founded at the Wistar Institute (USA)

The highly qualified personnel, scientific, methodological, technical and experimental base (BCL, vivarium) allow IEPOR to perform comprehensive preclinical testing of medicines and pharmaceutical substances as well as medical products that is, to provide relevant appropriate scientific and methodological level of preclinical study of safety and efficacy of medicines and the proper adherence of the principles of humane handling of laboratory animals in case they are used.

The tests are carried out according to the requirements of the Basic Principles of Good Laboratory Practice (GLP). Written protocol (plan) of preclinical studies is obligatory. In preclinical studies appropriate mathematical and statistical methods are used in order to ensure an adequate evaluation of the obtained results. The report of preclinical trials results of medicinal product is submitted to the Customer. All materials of the preclinical study of the medicinal product drug are archived for at least five years after completion of the preclinical testing.

Opportunities

The following types of preclinical tests can be conducted at IEPOR:

  1. Screening of bioactive substances and experimental study of toxic effects of medicinal products
  2. Study of mutagenic and carcinogenic properties of new substances and drugs
  3.  Preclinical studies of antitumor drugs
  4.  Preclinical study of new drugs
  5.  Study of the immunotoxic action of medicinal products

Namely:

Study of physical and chemical properties of medicinal products

The study of the physical and chemical properties of individual active substances (preformulation study) is carried out to identify the critical characteristics of the source materials that affect the quality of the finished product: determining the solubility of the biologically active substance or its salts in water and other solvents; study of its chemical stability in the dissolved and solid state; determination of the dependence of solubility and chemical stability on the dissociation constant and pH; studies of substances’ hygroscopicity, crystals’ polymorphism. The most suitable pharmaceutical substance for use in such preparations should have the following properties: high solubility in water; chemical stability; significant therapeutic width of action.


Stability in time. Storage conditions

The criterion for the stability of a medicinal substance is the preservation of its quality, that is, appearance, solubility, authenticity, high quality and quantitative content. Reducing the quantitative content of the pharmacologically active substance in the medicinal product confirms its instability. The toxic degradation products and/or changes in the physicochemical properties of the medicinal substance should not be formed. It is possible to determine the stability by the classic method and by the method of «accelerated aging». Analyzing the physical and chemical changes of the sudstance, one should assess its stability.


Acute toxicity

Determining the toxicological characteristics of a substance is the first stage of all toxicological tests, the purpose of which is to obtain information on danger in terms of short-term action. Assessment of the dosage regimes in the studies of subacute, chronic, specific toxicity and determination of the basic parameters of substances’ toxicity — the average lethal dose and its standard error (LD50 ± m), the maximum tolerated dose (DL0), as well as DL16 and DL84 are carried out.. Clinical signs of intoxication and pathomorphological studies are monitored and the relative mass of organs is determined.


Repeated dose toxicity

Finding out of subacute (substance toxicity due to administration for a limited time), subchronic (adverse effects of repeated injections over a certain part of life) and chronic toxicity (determination of safety limits of doses recommended for use). These studies are performed in three doses at two types of experimental animals of both sexes. The clinical, pathomorphologic, hematologic observations, and the organs’ mass determinations, biochemical analysis of serum and urine are performed.

Cumulative toxicity

The studies of toxicokinetics (pharmacokinetics) including absorption, distribution, transformation and excretion are taking place. Also we study the cumulativeness of the effect under consideration, i.e. definition of the average total amount of product received by the animal in subacute experiment prior to the appearance of certain effect and comparing this amount with a single average effective dose with the calculation of the cumulative coefficient.


Specific (antitumor) activity

Determining of cytostatic and cytotoxic action of the substance is performed in model systems in vitro and partly in vivo on available models of tumor growth. Positive results make it possible to study the antitumor activity on signaling compulsory obligatory models of experimental tumors (2-4strains), followed by advanced study of the investigated compound in a wide range of tumors (antitumor and antileukemia effect) — studies of dose dependence, of optimal ways and modes of application, as well as the selectivity of its action.


Morphological structure of internal organs; general biochemical and blood parameters

The macroscopic examination, organs’ weight, investigation of the histological structure of organs and tissues with obvious macroscopic changes, and microphotograph of histological preparations are performed. In toxicological tests the following organs: adrenal glands, brain, heart, kidneys, liver, lung, ovaries/testicles, pituitary gland, prostate/uterus, spleen, thymus are examined.


Genotoxic effects

We study: mutagenic potential of chemical compounds to identify potential mutagens and assess their genetic activity in the Ames test, to determine single-strand DNA breaks, to evaluate the frequency of gene mutations in cell cultures of different origin and to estimate chromosomal aberrations in mammalian cells in vitro and in vivo.


Side effect after the treatment

Morphological structure of the liver, kidneys, spleen, heart, lungs, bone marrow; biochemical and general blood parameters are assessed.


Studying carcinogenic properties of drugs and substances

The detection of substances as carcinogens — the ability to cause malignant tumors is studied. We conduct research on possible blastomogenicity on two kinds of animals of both sexes (mice and rats with low frequency of spontaneous tumors) using two doses of the substance. The pathomorphological evaluation of experimental material, histological examination, determination of life expectancy of animals with tumors, changes of animals’ mass, latent period of tumors’ emergence duration, etc. are carried out.


Testing for immunotoxicity

Studies of the non-specific link of the immune response and the two main groups of reactions — humoral and cellular type are performed. To do this, we use research methods that reflect the integral characteristics of the immune system links. Mature mice (S57Bl and CBA) are used.

New agents, the drugs

Examples of the IEPOR positive experience in preclinical trials of new substances and medicines:

BLASTEN

Glycopeptide from the cell wall of Lactobacillus delbrueckii 95% teyhoyevi acids, nucleotides, peptides <5%
Preclinical toxicological and pharmacological studies of the substance and dosage form, clinical trials were conducted


Ceruloplasmin

 Copper-containing enzyme of alpha-2-globulin fraction of serum from human donor’s serum.
The drug preparation with experimental and clinical studies have been carried out


BIOTSERULIN (ceruloplasmin)

Copper-containing enzyme of alpha-2-globulin fraction of serum from human donor’s serum.
Complex preclinical trials during drug re-registration were performed


5-AMINOLEVULENIC ACID

5-amino-4-oxopentanoic acid hydrochloride.
Antitumor photodynamic activity study was conducted


IDARUBICIN

Antitumor anthracycline antibiotic of the second generation, in which the hydrogen atom C4 is replaced by the mitoxyl group of aglycone.
Preclinical toxicological and pharmacological studies of dosage form were carried out


DOXORUBICIN

5,12-naftazenedion,
10-(3-amino-2,3,6-trideoxy-alpha-L-hexopyranosilyl)oxy)-7,8,9,10-tetrahydro-6,8,11-trihydroxy-8-(hydroxylacetyl)-1-(trifluoroacetyl)methoxyhydrochloride(8S-cis)
Preclinical toxicological and pharmacological studies of dosage form were carried out


 LIPODOX

Doxorubicin liposomal form.
Preclinical studies of toxicity and antitumor activity compared with doxorubicin for injections, and clinical trials have been performed


LAFERON

Recombinant human interferon alfa-2b.
Complex pre- and clinical trials have been performed


ENOANT

Food concentrate of grape polyphenols. Polyphenols of grape in the food concentrate "Enoant" are represented by anthocyanins, leucoanthocyanins, catechins, tannins in dissolved form. The total polyphenol concentration is 18-20 g/dm.
Experimental studies of antitumor activity and specific pharmacological action were performed


MEBIFON

(8S,10S)-10-[(3-amino-2,3,6-trihydroxy-a-L-arabino-hexopyranosyl)oxy]-8-glycoloyl-7,8,9,10-tetrahydro-6,8,11-trihydroxy-1-methoxy-5,12-naphthacenedione hydrochloride.
Clinical studies were performed

TROPISETRON

[(aH,5aH)-8-methyl-8-azobicyclo[3,2,1]oct-3a-yl]-1H-indole-3-carboxylate hydrochloride.
Preclinical toxicological and pharmacological studies of dosage form were carried out


EPIRUBICIN

(8S,10S)-10-[(3-amino-2,3,6-trihydroxy-a-L-arabino-hexopyranosyl)oxy]-8-glycoloyl-7,8,9,10-tetrahydro-6,8,11-trihydroxy-1-methoxy-5,12-naphthacenedione hydrochloride.
Preclinical studies of toxicity and antitumor activity in comparison with pharmorubіcin(epurabicin) of «Pharmacia» company were performed for the development of Ukrainian domestic generic drug


MITOXANTRONE

1,4-dihydroxy-5,8-bis(2-((2-hydroxyethyl)amino)ethyl)amino)anthraquinone dihydrochloride.
Preclinical toxicological and pharmacological studies of dosage form were carried out


ANTITUMOR AUTOVACCINE

 Vaccines designed on the basis of lectin isolated from culture of B. subtilis B-7025 and autologous tumor tissue of the patient.
Preclinical and clinical studies have been conducted


FEROPLAT

Antitumor nanocomposite is a novel form of the drug «Cisplatin».
Complex preclinical studies were carried out

Preclinical studies

Preclinical studies of medical products:

  1. Medical product «Dressing means based on fibrous carbon materials» for usage in the application and sorption therapy
  2. Biologically active supplement «CARBOLINE» — granulated carbon enterosorbent
  3. Medical product «Hemosorbent granulated deligandising (HSGD)» for deep purification of blood plasma proteins and blood cells membranes’ from endo- and exogenous substances of lipophilic and amphiphilic nature
  4. Monoclonal antibodies produced by IEPOR for the diagnosis of malignant tumors of different origin and evaluation of the immunological status
  5. Biologically active supplement «DONOVIT-VS2» is aconitine-containing agent of plant origin with antitumor and antimetastatic effect on malignant tumors with angiogenesis-dependent growth

 

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