Transporter-Targeted Drug Delivery

01.09.2012 -

Research group

Endogenous solute carriers (SLCs) and ATP-binding cassette (ABC) proteins transport essential substrates, such as nutrients, hormones, neurotransmitters, metabolites, and drugs across the cell membranes and therefore, have a crucial role in substances’ distribution and disposition in the body. These transporters can also be utilized to enhance drug delivery and target drugs to specific organs of cells. Moreover, if transporter function or expression can be modulated in specific cells, the drug efficacy and safety could be greatly improved.

Transporter-mediated Drug Delivery and Targeting

Our research group studies how transporter proteins can be utilized in order to target drugs to their site of action, which would improve their efficacy and safety. We develop novel transporter-utilizing compounds and prodrugs that can release the active drug at their target site. More specifically we aim to:

1. Improve the drug delivery across the blood-brain barrier (BBB) and selectively into different cell types (neurons, astrocytes or microglia) within brain (intra-brain targeting)

  • Explore if dual targeting of LAT1-utilizing compounds into the brain and simultaneously into the pancreas (brain-pancreas axis) can have beneficial effects on the treatment of neurodegenerative diseases

2. Target chemotherapeutics (prodrugs or drug analogues) selectively into the cancer cells

3. Inhibit specific transporters (influx or efflux) to block nutrient delivery into the cancer cell or drug efflux out of the cancer cells (chemoresistance) to increase antiproliferative effects

4. Modulate nutrient uptake (e.g. amino acids) or toxin uptake into the brain that can affect the progression of neurological diseases

5. Down-regulate transporter expression selectively in cancer cells or up-regulated transporter expression in target organs, such as brain or pancreas

 

We follow sustainable research ethics and support personal goals in lifelong learning pathway.

We discuss via open science and social interactions in the community.

Our multidisciplinary group promote equality and non-discrimination.

 

Homepage of research group

Keywords

Time period

01.09.2012 -

    Our Research Projects

    • Intra-Brain Targeting

      +

      Solute Carriers (SLCs) are highly and selectively expressed in the brain in relation to other healthy tissues. For example, L-type Amino Acid Transporter 1 (LAT1), Organic Anion Transporting Polypeptides (OATPs), Monocarboxylate Transporters (MCTs), Organic Cation Transporters (OCTs) and Organic Anion Transporters (OATs) are highly and/or selectively expressed on the blood-brain-barrier, and in certain brain cells, such as neurons, astrocytes, and microglia or in activated immune cells. Therefore, those transporters can be utilized for brain-targeted as well as intra-brain targeted drug delivery.

      The main aim of our research group is to study the utilization of these transporters in:

      1. Targeted (pro)drug delivery of neuroprotective agents across the BBB
      2. Targeted (pro)drug delivery into target cell types within the brain

      The research in this area is expected to increase the efficacy of drug therapies related to central nervous system (CNS)- diseases due to the increased drug accumulation to the desired tissues and cells. However, it is expected to decrease simultaneously unpleasant adverse effects that arise from unselective drug delivery and the distribution of drugs to off-target organs. We are especially interested in CNS-diseases, in which neuroinflammation is related, such as neurodegenerative disorders (such as Alzheimer’s Disease, Parkinson’s Disease, Multiple Sclerosis, etc.), stroke and acute brain infections.

    • Cancer Cell Targeting

      +

      Solute Carriers (SLCs) as well as ATP-binding cassette (ABC) transporters, so-called efflux transporters are over-expressed in many different types of cancer cells and in solid tumors. Thus, SCLs can be utilized for cancer cell-targeted selective drug delivery or to inhibit and starve the cancer cell from essential nutrients. ABC transporters, on the other hand, have a crucial role in cancer biology and more importantly in chemoresistance, as they can pump a wide range of chemotherapeutics out of the cancer cells and thereby reduce the efficacy of these anti-cancer drugs. Therefore, targeting efflux transporter inhibitors selectively into the cancer cell may offer a potential alternative to improve the efficacy of chemotherapeutics.

      The main aim of our research group is to study:
      1. LAT1 as a Selective Carrier of Chemothrerapeutics or Efflux Transporter Inhibitors
      2. Other Transporters as Novel Carriers of Anti-Cancer Agents
      3. LAT1 Inhibition to Prevent Cancer Cell Proliferation

      By inhibiting L-type amino acid transporter 1 (LAT1) that is over-expressed in several cancer types, protein synthesis and cell proliferation and can be hindered. We have reported that this inhibition is very effective alone, but it can also potentiate the effects of several other anti-cancer agents. This is expected to improve not only the efficacy but also the safety of chemotherapy, as the amounts of used anti-cancer agents can be reduced. More importantly, a selective inhibitor can be developed also as a novel diagnostic agent and be studied as a modifying tool for brain amino acid homeostasis. Although we are currently studying mainly LAT1 as a carrier and a target protein, we are constantly looking up also other potential SCLs, such as organic cation transporters (OCTs)in the delivery of metformin derivatives.

    • Pancreatic Targeting

      +

      Pancreas and pancreatic cells are very heterogeneous, which makes the drug delivery into the pancreas and e.g., insulin-secreting pancreatic β-cells very challenging. We have recently noticed that our L-type amino acid transporter 1 (LAT1)- utilizing prodrug can accumulate very effectively into the pancreas. Therefore, we are interested in exploring

      1. if LAT1-utilizing prodrugs could be used to treat pancreas-related diseases, not only pancreatic cancer, but also pancreatitis and diabetes

      2. dual effects of LAT1-utilization, both in brain and pancreas to see if there is any connection between these organs, in diabetes-related neurodegenerative diseases, like Alzheimer’s disease

    • Peiotropic Metformin Derivatives (in Collaboration)

      +

      Numerous studies have shown that type 2 diabetes (T2DM) is associated with thrombosis and the activation of blood coagulation. In addition, the diabetic state is characterized by an impaired balance between the processes of coagulation and fibrinolysis, known as diabetic thrombophilia. Most newly-diagnosed patients with T2DM are placed on metformin, an oral glucose-lowering drug, which also has beneficial effects on the cardiovascular system. Metformin can positively affect endothelial function, protect from oxidative stress and inflammation, and inhibit the proliferation of smooth muscle cells.

      Our research collaboration with Dr. Magdalena Marcowicz-Piasecka at the Medical University of Lodz (Poland) aims to evaluate the effects of metformin and its novel sulfenamide and sulfonamide derivatives produced at the UEF on selected parameters of haemostasis, including:

      1. Plasma Haemostasis (e.g., overall haemostasis potential, extrinsic and intrinsic coagulation pathway, activity of coagulation factor)

      2. Vascular Function (e.g., function, viability, and adherence of endothelial cells, the release of coagulation factors from ECs, the proliferation of smooth muscle cells)

      3. Platelets Haemostasis (e.g., platelet aggregation, adhesion, thrombus formation in semi-physiological conditions)

      Metformin has also been reported to play an important role in the treatment of Alzheimer’s Disease (AD). Therefore, our secondary aim is to assess the effects of metformin and its novel sulfenamide and sulfonamide derivatives on the activity of human AChE and BuChE and to establish the type of inhibition to provide greater insight into the more rational design of potential cholinesterase (ChE) inhibitors with a biguanide skeletal structure. Moreover, metformin has also found to act synergistically with donepezil, which might have potential in preventing brain disorders associated with diabetes complications in the future or might provide an additional rationale for the clinical benefit for AD subject suffering simultaneously from T2DM. Therefore, novel metformin derivatives are also explored for their potential synergism with other clinically approved anti-AD drugs.

      We are also trying to identify the possible transport mechanisms of novel metformin derivatives, and build up structure-activity relationships (SARs), as it is already known that metformin can interact with organic anion transporters 1-3 (OCT1-3), multidrug toxin and extrusion 1-2 (MATE1-2), plasma membrane monoamine transporter (PMAT).

    Methods

    • Read More About Our Methodology

      +

      Computational drug design is used in the design of transporter-utilizing (pro)drugs and transporter inhibitors.

      The designed prodrug molecules are synthesized, purified (recrystallization, chromatography), and characterized (NMR, MS, Elemental analysis) in our state-of-the-art synthesis laboratories.

      Physicochemical and pharmaceutical properties (e.g. distribution coefficient (log D), aqueous solubility) and chemical stabilities at various pH values as well as plasma protein binding of the novel prodrugs are determined in our state-of-the-art analysis laboratories. The rate and quantity of the released parent drugs are evaluated in vitro in various enzyme-containing media (rodent and human-derived) and buffered solutions containing pure enzymes.

      Transporter- mediated uptake rate and extend of (pro)drugs into the human cells are evaluated in suitable cell lines expressing the desired transporter. The analysis are carried out in updated analytical equipment (UPLC-DAD, HPLC-F or LC-MS/MS) after suitable sample preparation.

      (Pro)drug pharmacokinetics and targeting efficacies are evaluated with more sophisticated in vivo studies in rodents. The analyses are carried as mentioned above and metabolic profiles of novel compounds are studied . Finally, in vitro-in vivo correlations of compound structure and targeting efficacy are projected.

      Pharmacological efficacy (preventative and curative) and possible toxicological properties (biocompatibility, cytotoxicity, genotoxicity, etc.) of the developed (pro)drugs are studied firstly in vitro and secondly in vivo. Reliable biomarkers (already known but also possible novel biomarkers) are identified by LC-MS/MS (targeted/global proteomics/metabolomics). Finally, in vitro-in vivo correlations are projected and translated to human situation.

    Vacancies

    Postdoctoral Candidates
    Unfortunately, we do not have currently any open positions for postdoctoral researchers. However, our group is happy to support and help in writing any candidate with a clear vision of the proposed project that could be funded by e.g., Marie Skłodowska-Curie Individual Fellowship, Academy of Finland Postdoctoral Fellowship, UEF Postdoctoral Fellowship, or any other grants from national or international foundations.

    Topics for M.Sc. Thesis or Diploma Work in Pharmacy for ERASMUS students:

    • 1. Molecular modelling and synthesis of neuroprotective or anti-cancer agents

      +
      • Methods: Computer-aided drug design, Docking and Synthesis (organic chemistry) and Structural Characterization (NMR, MS, elemental analysis) of Designed Molecules
      • Main supervisor: Adj. Prof. Kristiina Huttunen, Ph.D. and Dr. Maija Lahtela-Kakkonen, Ph.D.
      • Contact: kristiina.huttunen (at) uef.fi
      • Other information: In this project, the student will design novel compounds against neurodegenerative diseases or cancer by molecular modelling and then synthesize the molecules. Prepared compounds are then purified by chromatographic or crystallographic methods and finally characterized by NMR (nuclear magnetic resonance) and mass spectroscopy as well as by elemental analysis. The compounds are studied further against neurodegenerative diseases or cancer in our in vitro and in vivo laboratories. The prepared compounds depends on the current state of the research and selected target protein. In this project, the student will learn both computer-aided drug design and well as to perform essential methods in organic chemistry. PROMM course is mandatory for modelling.
    • 2. Design and synthesis of brain-targeted transporter-utilizing prodrugs of neuroprotective agents used against neurodegenerative diseases

      +
      • Methods: Design (database), Synthesis (organic chemistry) and Structural Characterization (NMR, MS, elemental analysis) of Drug Molecules
      • Main supervisor: Adj. Prof. Kristiina Huttunen, Ph.D. and Dr. Santosh Adla, Ph.D.
      • Contact: kristiina.huttunen (at) uef.fi
      • Other information: In this study, novel designed prodrugs of neuroprotective agents are synthesized and purified by chromatographic or crystallographic methods and finally characterized by NMR (nuclear magnetic resonance) and mass spectroscopy as well as by elemental analysis. In this project, the student will learn to design multistep reaction routes by using several databases and published literature as well as to perform essential methods in organic chemistry. Prepared prodrugs are then studied further against neurodegenerative diseases in our in vitro and in vivo laboratories. The project requires basic knowledge of organic/synthetic chemistry. However, the subprojects are scalable to both to Master Thesis level as well as to ERASMUS students. The prepared compounds depend on the current state of the research.
    • 3. Bioconversion, brain-cell selective pharmacodynamic studies of transporter-utilizing prodrugs of neuroprotective agents used against neurodegenerative diseases

      +
      • Methods: Chemical and enzymatic stability, solubility, unspecific protein binding, HPLC (UV/MS), targeted/global proteomics
      • Supervisors: Adj. Prof. Kristiina Huttunen, Ph.D., Janne Tampio, M.Sc., and Ahmed Montaser, M.Sc.
      • Contact: kristiina.huttunen (at) uef.fi
      • Other information: In this project, physicochemical and pharmaceutical properties, such as aqueous solubility, chemical stability, unspecific plasma/tissue protein binding, enzymatic bioconversion in various biological matrices (plasma, microsomes, liver/brain homogenate) and/or with pure enzymes of novel prodrugs of selected neuroprotective agents synthesized in our laboratories are evaluated. For each prodrug a simple HPLC (High Performance Liquid Chromatography)  or LC-MS (Mass Spectrometry) method are developed, by which the samples are analyzed. Selected biomarkers for each neuroprotective parent drugs and their prodrugs are evaluated by MS with targeted or untargeted (global) proteomic/lipidomic methods. In this study, the student will learn basic pharmaceutical techniques and learn to analyze properties of compounds from various sample matrices. The project requires basic knowledge of analytical chemistry. However, the subprojects are scalable to both to Master Thesis level as well as to ERASMUS students. The prepared compounds depend on the current state of the research.
    • 4. Cancer cell-selective transporter-mediated uptake studies and anti-proliferative efficacy of transporter inhibitors and their prodrugs

      +
      • Methods: Cell culturing, cytotoxicity, HPLC (UV/MS)
      • Supervisors: Adj. Prof. Kristiina Huttunen, Ph.D., Janne Tamöpio, M.Sc., and Johanna Huttunen, M.Sc.
      • Contact: kristiina.huttunen (at) uef.fi
      • Other information: In this project, cellular uptake of transporter inhibitors and their effects on chemo-resistance mediated by efflux transporters are evaluated in human breast cancer cell lines. Transporter inhibitors and their cancer-cell targeted prodrugs are studied together with selected anti-cancer agent to evaluate the increase of combination therapy in anti-proliferative efficacy. In this study, the student will learn to handle the cell line, to perform the uptake and cell viability studies as well as analyze the results by a feasible HPLC (High Performance Liquid Chromatography) methods, which will be developed for each prodrug. The project requires basic knowledge of analytical chemistry and cell culturing. However, the subprojects are scalable to both to Master Thesis level as well as to ERASMUS students. The prepared compounds depend on the current state of the research.
    • 5. In vitro affinity and cellular uptake of LAT1-utilizing prodrugs/compounds

      +
      • Methods: cell culturing, working with radiolabeled materials, HPLC (UV/MS)
      • Supervisors: Adj. Prof. Kristiina Huttunen and Ph.D., Janne Tampio, M.Sc., Juulia Järvinen, M.Sc., or M.Sc., Johanna Huttunen, M.Sc.
      • Contact: kristiina.huttunen (at) uef.fi
      • Other information: In these studies, in-house prepared (pro)drugs are evaluated as substrates for L-type amino acid transporter 1 (LAT1). This includes cis- and trans-inhibition studies of radiolabeled amino acids as well as concentration-dependent cellular uptake of studied compounds in human cancer cells or brain cells (e.g., microglia or astrocytes) over-expressing LAT1 or LAT1-tranfected cell line. The student will learn to handle the cell line as well as to perform the uptake studies and analyze the results by a feasible UV- or MS- HPLC (High Performance Liquid Chromatography) method, which will be developed for each prodrug. Several sub-projects are available and the studied compounds depend on the current state of the research. The project requires basic knowledge of analytical chemistry and cell culturing. However, the subprojects are scalable to both to Master Thesis level as well as to ERASMUS students.

       

    • 6. Pharmacokinetic and comparative efficacy and safety studies of neuroprotective agents and their brain-targeted prodrugs in neuroinflammation-induced mice

      +
      • Methods: in vivo, LC-MS, targeted/global proteomics
      • Supervisors: Adj. Prof. Kristiina Huttunen, Ph.D. and Ahmed Montaser, M.Sc.
      • Contact: kristiina.huttunen (at) uef.fi
      • Other information: In this project, pharmacokinetic properties and brain-targeting efficacy of novel in-house synthesized prodrugs of neuroprotective agents are evaluated from the samples collected after in vivo studies from mice. Furthermore, selected biomarkers are followed from neuroinflammated mice induced by lipopolysaccharide (LPS) by targeted or untargeted (global) proteomic approach with mass spectrometry (MS). The student will learn essential sample preparation techniques (extraction, protein precipitation) of plasma and tissue samples (brain, liver, kidney, pancreas, etc.) and to analyze compounds and biomarkers from biological matrices by feasible LC-MS methods. If wanted, student can also participate in in vivo studies. The project requires basic knowledge of analytical chemistry and possibly animal experiments. However, the subprojects are scalable to both to Master Thesis level as well as to ERASMUS students. The studied compounds depend on the current state of the research.
    • 7. Analytical method development in targeted /untargeted proteomic data analysis

      +
      • Methods: Mass spectrometry, targeted/untargeted proteomics, (pathway) data analysis
      • Supervisors: Adj. Prof. Kristiina Huttunen, Ph.D., and Ahmed Montaser, M.Sc.
      • Contact: kristiina.huttunen (at) uef.fi
      • Other information: In this project, transporter expression levels are explored by targeted proteomic approach from different cell or tissue samples to understand endogenous and exogenous compounds’ disposition in healthy and disease stages. In addition, global proteomic approach (untargeted) and pathway analysis will be used to understand detailed mechanisms behind different diseases. In this study, the student will learn essential proteomic sample preparation techniques and how to analyze a massive amounts of data. The project requires basic knowledge of analytical chemistry and proteomic approach. However, the subprojects are scalable to both to Master Thesis level as well as to ERASMUS students. The exact topic depend on the current state of the research. The exact topic depend on the current state of the research.

    Funding

    • Thank You for the Scientific Support!

      +

      Academy of Finland

      Sigrid Juselius Foundation

      Päivikki and Sakari Sohlberg’s Foundation

      Jane and Aatos Erkko Foundation

      Magnus Ehrnrooth Foundation

      UEF Strategic Funding (Doctoral Programme and Infrastructure Investments)

      Finnish Cultural Foundation

      Emil Aaltonen Foundation

      Alfred Kordelin Foundation

      Orion Research Foundation

      Finnish Pharmaceutical Society

      Paavo Koistinen Foundation

Group members - UEF

Other group members

  • Katayun Bahrami
    Katayun Bahrami katayun.bahrami@uef.fi
  • Juulia Järvinen
    Juulia Järvinen juulia.jarvinen@uef.fi
  • Arun Tonduru
    Arun Tonduru arun.tonduru@uef.fi
  • Santosh Adla
    Santosh Adla santosh.adla@uef.fi

Links

Publications by the research group members