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Respiratory activity

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Drug Discovery and Evaluation

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1 D.1 In vitro tests

1.1 D.1.0.1 Histamine (H1) receptor binding

1.1.1 PURPOSE AND RATIONALE

Histamine is considered to play a major role in asthmatic attacks. H1-antagonists have been used since decades as therapeutic agents. This assay is used to determine the affinity of test compounds to the histamine H1 receptor by measuring their inhibitory activities on the binding of the H1 antagonist 3H-pyrilamine to a plasma membrane preparation from guinea pig brain.

1.1.2 PROCEDURE

Brains from guinea-pigs are homogenized in ice-cold Tris buffer (pH 7.5) in a Potter homogenizer (1 g brain in 30 ml buffer). The homogenate is centrifuged at 4°C for 10 min at 50000g. The supernatant is discarded, the pellet resupended in buffer, centrifuged as before, and the final pellets resupended in Tris buffer (1 g fresh weight/5 ml). Aliquots of 1 ml are frozen at −70°C.

In the competition experiment, 5 μl 3H-pyrilamine (one constant concentration of 2×10−-9 M), 50 μl test compound (>10 concentrations, 10

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Authors and Affiliations

  1. Johann Wolfgang Goethe-Universität, Frankfurt, Germany

    Prof. Dr. Dr. H. Gerhard Vogel

  2. Department of Pharmacology, Jefferson Medical College, Thomas Jefferson University, Philadelphia, USA

    Prof. Dr. Wolfgang H. Vogel

  3. Aventis Deutschland GmbH, Johann Wolfgang Goethe-Universität Frankfurt, Frankfurt, Germany

    Prof. Dr. Bernward A. Schölkens & Prof. Dr. Jürgen Sandow

  4. Institut für Genetik, Avenfis Deutschland GmbH, Ludwig Maximilian-Universität München, München, Germany

    PD Dr. Günter Müller

  5. Department of Laboratory Medicine and Pathobiology, Faculty of Medicine, University of Toronto, Toronto, Canada

    Dr. Wolfgang F. Vogel (Assistant Professor)

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  1. Johann Wolfgang Goethe-Universität, Frankfurt, Germany

    H. Gerhard Vogel

  2. Department of Pharmacology, Jefferson Medical College, Thomas Jefferson University, Philadelphia, USA

    Wolfgang H. Vogel

  3. Aventis Deutschland GmbH, Johann Wolfgang Goethe-Universität Frankfurt, Frankfurt, Germany

    Bernward A. Schölkens  & Jürgen Sandow  & 

  4. Institut für Genetik, Avenfis Deutschland GmbH, Ludwig Maximilian-Universität München, München, Germany

    Günter Müller

  5. Department of Laboratory Medicine and Pathobiology, Faculty of Medicine, University of Toronto, Toronto, Canada

    Wolfgang F. Vogel (Assistant Professor) (Assistant Professor)

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Vogel, H.G., Vogel, W.H., Schölkens, B.A., Sandow, J., Müller, G., Vogel, W.F. (2002). Respiratory activity. In: Vogel, H.G., Vogel, W.H., Schölkens, B.A., Sandow, J., Müller, G., Vogel, W.F. (eds) Drug Discovery and Evaluation. Springer, Berlin, Heidelberg. https://doi.org/10.1007/3-540-29837-1_5

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