Discovery Pipeline
  • Development of aldosterone synthase inhibitors

    Aldosterone synthase inhibitor is one of the most anticipated drug in cardio-metabolic therapy. We develop compounds that selectively inhibit synthesis of aldosterone for future use in therapy of drug-resistant arterial hypertension and non-genomiс effects of high aldosterone. We are the first to fulfill this gap by solving the crystal structure of aldosterone synthase in complex with substrate and inhibitor.

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  • Anti-tuberculosis drug discovery

    We’ve identified and validated a new targets – mycobacterial cytochromes P450, responsible for cell wall formation and depression of local immune response in human. This achievement is a basis for a drug discovery for treatment of resistant (MDR, XDR) forms of tuberculosis. 

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  • Potential for a future pipeline

    Rare diseases, fungal infections and cancer are an examples of our potential to extend our pipeline future.

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Development of aldosterone synthase inhibitors

Inhibitors of aldosterone synthase

Development of selective compounds that exclusively inhibit aldosterone synthesis fir use in therapy of resistant arterial hypertension caused by hyperaldosteronism. Currently there is no drug on the market targeting aldosterone synthase. We are the first to fulfill this gap by solving the crystal structure of aldosterone synthase in complex with substrate and inhibitor. We've identified 4 scaffolds for further development and undergoing the HTS.

According to the latest WHO data, more than 7.5 million deaths worldwide are caused by high blood pressure (hypertension), representing approximately 12.8% of total deaths. The sum of years lost due to the premature death and disability (indicator DALYs) due to hypertension is 58 million. Special attention is given to resistant forms of hypertension (RFH), when the combination of three or more antihypertensive drugs of different classes cannot eliminate the symptoms of hypertension. Patients with RFH constitute 5-30% of patients with hypertension and have more severe symptoms. 

To a large extent the progression of RFH is related to the excessive aldosterone synthesis (basic human mineralocorticoid hormone). Aldosterone excess is implicated in the development and progression of several different cardiovascular/renal disease processes, including hypertension, congestive heart failure, chronic kidney disease, coronary artery disease and stroke. High aldosterone concentrations have also been associated with insulin resistance and pancreatic ?-cell dysfunction. Most prominent approach to inhibiting aldosterone-induced cardiovascular injury is to suppress its synthesis (i.e., the inhibition of aldosterone synthase). This strategy prevents the non-genomic actions of aldosterone that are not antagonized by MR blockade. Therefore aldosterone synthase (CYP11B2 or AS) has received growing attention as an attractive therapeutic target for antihypertensive drug discovery.

However, the development of selective and potent CYP11B2 inhibitors is particularly challenging due to high homology to the CYP11B1 isoform (93% amino acid sequence identity with CYP11B2) and other steroidogenic enzymes, especially in the absence of structural information. The progress in AS research was hampered for a long time by the inability to obtain a purified protein due to its low stability and membrane nature and as a result no adequate screening systems available.

We use two alternative approaches in the design of aldosterone synthase inhibitors: 1) screening of more specific inhibitors of the enzyme’s active site (based on our crystal structures of human aldosterone synthase in complex with substrate and inhibitor (PDB ID: 4FDH and 4FDH); 2) screening of peptidomimetics that prevent binding of the redox partner to the mitochondrial P450 (based on our crystal structure of the mitochondrial CYP11A1 in complex with redox partner - adrenodoxin).

For both approaches purified protein expressed in E.coli is used. Protein production is according to our developed and published protocol (Strushkevich N et al. Structural insights into aldosterone synthase substrate specificity and targeted inhibition. Mol Endocrinol. 2013). Availability of purified protein has allowed us to develop and implement a high-throughput  screening for testing of potential inhibitors and as a result improve techniques used in other laboratories, e.g. detecting of aldosterone synthase activity in reconstructed system using: a) an enzyme expressed at a low level in eukaryotic cells; b) the radioactive substrate. To obtain a selective inhibitor all experiments will be carried out in tandem with two proteins, CYP11B1 and CYP11B2 (93% homology). In addition, project owners have a number of steroidogenic P450, as well as human liver cytochromes P450, which will be tested for cross-inhibition.

Currently the high- throughput screening of aldosterone synthase inhibitors enabled to identify four different types of compounds with strong inhibitory effect on aldosterone synthase. These compounds will be used for molecular docking using the crystal structure of the complex with fadrazole and for subsequent optimization using small fragments library. A high- throughput screening will be carried out in silico and in vitro using commercial library of chemical compounds of various classes and mechanisms of action (1.5 mln compounds). Obtained hit/s will be used for assessment of inhibition effects in reconstructed system and for IC50 detrmination. We are planning to obtain the crystal structure in complex a hit compound for structure-based drug design and for further optimization.

The design of peptidomimetic will be carried out based on the data about the contact zone of the protein partners, according to the crystal structure of P450 and adrenodoxin. The synthesis will be carried out using an automated peptide synthesizer Intavis utilizing both natural and artificial/modified amino acids. In addition, phage-display technology will be used for the screening of peptides having high affinity for aldosterone synthase.

Our compound will have a high inhibitory activity on a specific target (aldosterone synthase), have a minimal inhibitory effect on other enzymes of the steroid hormones biosynthesis and xenobiotic metabolism, and also have low toxicity. 

Anti-tuberculosis drug discovery

Anti-tuberculosis drug discovery

Key facts (from WHO)
  • Tuberculosis (TB) is one of the top 10 causes of death worldwide.
  • In 2015, 10.4 million people fell ill with TB and 1.8 million died from the disease (including 0.4 million among people with HIV). Over 95% of TB deaths occur in low- and middle-income countries.
  • In 2015, an estimated 1 million children became ill with TB and 170 000 children died of TB (excluding children with HIV).
  • TB is a leading killer of HIV-positive people: in 2015, 35% of HIV deaths were due to TB.
  • Globally in 2015, an estimated 480 000 people developed multidrug-resistant TB (MDR-TB).  In some cases, more severe drug resistance can develop. Extensively drug-resistant TB (XDR-TB) is a more serious form of MDR-TB caused by bacteria that do not respond to the most effective second-line anti-TB drugs, often leaving patients without any further treatment options. Worldwide, only 52% of MDR-TB patients and 28% of XDR-TB are currently successfully treated. The cost to treat a single XDR-TB case is over €160 000.
The MDR-TB burden largely falls on 3 countries – China, India, and the Russian Federation – which together account for nearly half of the global cases. About 9.5% of MDR-TB cases had XDR-TB in 2015.
“You’re really looking at a global issue .. It’s not a foreign problem, you can’t keep these TB patients out. It’s time people realize that.”
Dr Lee Reichman  (Mendoza, M, “First U.S. case of extremely drug resistant strain of tuberculosis diagnosed”, Associated Press 2009 )

New effective drugs need to be developed to treat of drug resistant (MDR, XDR) forms of tuberculosis. We’ve identified and validates a new targets – mycobacterial cytochromes P450, responsible for cell wall formation and depress of local immune response in human. Our team is developing an effective approach to inhibit a panel of mycobacterial cytochrome P450s with a minimal effect on human enzymes.

New generation of TB-drugs (inhibitors of mycobacterial-P450s) will inhibit the formation of cell wall components (stop growth of mycobacteria and make them sensitive to available dugs), and decrease effects of mycobacterial enzymes on immune system of host organism (increase innate immunoresponse to M.tuberculosis).

Potential for a future pipeline

Drug-resistant infections and valuable side products

 

Novel drugs for antifungal treatment (azole resistant strains)

Target – sterol-14-demethylase (CYP51)

Market – 13.9 bln USD by 2018 globally

 

Fungal infections in humans range from the superficial and common, such as dermatophytoses and onychomycoses, to deeply invasive and disseminated, such as candidiasis and aspergillosis. In the past 20 years, the frequency of systemic fungal infections has increased dramatically along with the number of invasive, mostly opportunistic, species. The main factor for the increase is the proliferation if severely immunocompromised patients either with AIDS, undergoing cancer chemotherapy, or immunosuppressive therapy for organ transplantation. Additional factors include treatment with broad-spectrum antibacterial drugs or glucocorticosteroids; invasive procedures such as surgery, in-dwelling catheters or prosthetic devices; and parenteral nutrition or dialysis.

The major pathogen has been Candida albicans, normally a commensal of the oral cavity and gastrointestinal tract of humans. Non-albicans Candida species (e. g. C. glabrata, C. tropicalis, C. krusei), however, are also with increasing frequency, as is Aspergillus sp., Histoplasma capsulatum and Cryptococcus neoformans. Emerging oportunitistic pathogens include Fusarium and Trichosporon (both significant in neutropenic patients and commonly associated with disseminated infection), Rhizopus, Mucor, dematiaceous fungi, and others. Fungal infections are important causes of morbidity and mortality in hospitalized patients: candidiasis is the fourth most common blood culture isolate in US hospitals, pulmonary aspergillosis is the leading cause of death in bone marrow transplant recipient, and Pneumocystis carinii (formerly thout to be a protozoan parasite) pneumonia is the leading cause of death in AIDS patients in Europe and North America.

The increase in life-threatening fungal infections has brought about an increased use of antifungal drugs and a pressing need for new, broad-spectrum, fungicidal chemical agents that can be used empirically in immunocompromised patients. Empirical treatment of suspected fungal infections is necessitated by problems in diagnosis and susceptibility testing and may be more common than treatment of confirmed infections.

 

Novel drugs for Buruli ulcer & leprosy treatment

Target – P450 involved in mycolactone synthesis

Buruli ulcer accounts 87.7 cases per 100,000 in endemic countries

Leprosy accounts >20 cases per 100,000 in endemic countries

 

Novel drugs for Chagas disease & sleeping sickness treatment

Target – CYP51 of Trypanosoma cruzi and T. brucie

Chagas disease accounts 10,300 deaths/year (affects 8 to 10 million)

Sleeping sickness – 9,000 deaths/year (risk 70 mln/36countries)

 

Novel drugs for breast cancer treatment

Target – aromatase (CYP19A1)

Market – 2,8 bln USD in 2012

Threat – Lifetime risk of developing invasive breast cancer is 12% (1 in 8 women)

 

Novel drugs for prostate cancer treatment

Target – CYP17A1

Market – 1,07 bln USD in 2014

Threat – Lifetime risk of developing prostate cancer is 16.7% (1 in 6 men)

Target medicals

Breast cancer, prostate cancer, resistant hypertension, infectious diseases – are just few examples of the human P450s dysfunction. We are studying all aspects of P450 function not only in humans but in human pathogens as well. By applying a protein family-based approach in structure-functional studies of these proteins new selective drugs can be designed and developed to treat life-threatening diseases.Current directions in drug discovery: cardiovascular and infection diseases, endocrine disorders.