Inhibitors of membranous adenylyl cyclases

doi:10.1016/j.tips.2011.10.006

Trends in Pharmacological Sciences

Volume 33, Issue 2, February 2012, Pages 64-78

https://doi.org/10.1016/j.tips.2011.10.006 Get rights and content

Membranous adenylyl cyclases (mACs) constitute a family of nine isoforms with different expression patterns. Studies with mAC gene knockout mice provide evidence for the notion that AC isoforms play distinct (patho)physiological roles. Consequently, there is substantial interest in the development of isoform-selective mAC inhibitors. Here, we review the current literature on mAC inhibitors. Structurally diverse inhibitors targeting the catalytic site and allosteric sites (e.g. the diterpene site) have been identified. The catalytic site of mACs accommodates both purine and pyrimidine nucleotides, with a hydrophobic pocket constituting a major affinity-conferring domain for substituents at the 2′- and 3′-O-ribosyl position of nucleotides. BODIPY-forskolin stimulates ACs 1 and 5 but inhibits AC2. However, so far, no inhibitor has been examined at all mAC isoforms, and data obtained with mAC inhibitors in intact cells have not always been interpreted cautiously enough. Future strategies for the development of the mAC inhibitor field are discussed critically.

Section snippets

Membranous adenylyl cyclases (mACs)

ACs catalyze the conversion of ATP into the second messenger cAMP. cAMP plays a crucial role in the regulation of numerous cell functions. Mammals express nine mAC isoforms and a sAC. mACs consist of two transmembrane domains with six predicted helices each, and two cytosolic domains, referred to as C1 and C2, respectively 1, 2, 3. The C1 and C2 domains of mACs show considerable homology with each other and together constitute the catalytic core of the enzyme. The C1 and C2 domains are

Challenges to isoform-specific mAC inhibitors

AC inhibitors are divided into four classes: (i) inhibitors competing with the substrate ATP at the catalytic site [9]; (ii) non-competitive/uncompetitive inhibitors mimicking the cAMP^.PP_i transition state (P-site inhibitors) [10]; (iii) allosteric non-competitive inhibitors targeting the diterpene site [11]; and (iv) allosteric non-competitive inhibitors targeting as yet undefined sites [12]. Both the catalytic and diterpene sites are highly conserved among mAC isoforms (Figure 1). Thus, from

Exploring the catalytic site of mACs with MANT- and TNP-nucleotides

MANT-nucleotides have provided valuable structural information on the properties of the catalytic site of mACs. Four crystal structures of the purified catalytic mAC subunits VC1:IIC2 in complex with various MANT- and TNP-nucleotides have been resolved (Figure 2a and c) 23, 24, 25. This information provides an excellent basis for future development of non-nucleoside/nucleotide-based mAC inhibitors. The inhibitory potencies of MANT-nucleotides are catalysis-dependent (i.e. the higher the AC

Recent developments on P-site inhibitors

Although highly potent P-site inhibitors such as 2′,5′-dideoxy-3′-ATP have been described and mAC crystal structures with this ligand resolved (Figure 2b) 10, 15, 38, a major problem has been the lack of isoform-specificity of these compounds 10, 15. More recently, non-nucleoside P-site inhibitors with supposedly higher mAC isoform-specificity have been reported (Table 1). Patent activity on P-site inhibitors, in contrast to competitive inhibitors or diterpenes, has been substantial (Table 2),

The diterpene site as target for mAC inhibitors

Traditionally, diterpenes have been associated with stimulatory effects on mACs 4, 32. Early studies showed that ACs 1, 2 and 5 interact differently with diterpenes [33], and later studies documented different activation patterns of these ACs by diterpenes 11, 34. Fluorescence studies with MANT-GTP at VC1:IIC2 revealed that, in contrast to earlier assumptions [39], 1-deoxy-forskolin (1d-FS) and 1,9-dideoxy-forskolin (1,9dd-FS) bind to AC [40]. Unlike FS and 9-deoxy-forskolin (9d-FS), 1d-FS and

Remaining questions

There is great interest in obtaining selective AC5 inhibitors as potential drugs for treatment of heart failure and aging (Table 2) [6]. However, based on the high amino acid sequence similarities of the C1 and C2 domains, respectively, of mAC isoforms (Figure 1), pharmacological discrimination between ACs 5 and 6 is challenging, at least when the catalytic site is targeted [15]. Even if selective AC5 inhibitors can be developed, neurotoxicity constitutes a serious issue. Specifically, AC5

Future directions

Future studies aiming at the development of inhibitors targeting the catalytic site of ACs, by analogy to protein kinases [37], should consider non-nucleoside/nucleotide-based compounds. Such studies entail high-throughput screening and are a task for the pharmaceutical industry. mAC inhibitors, particularly if they specifically explore the hydrophobic pocket in the catalytic site (Figure 2a), should possess better membrane permeability than nucleosides/nucleotides.

In principle, it is possible

Concluding remarks

By combining methods from biochemistry, pharmacology, biophysics and medicinal chemistry, substantial progress has been made towards understanding the molecular basis of the interactions of inhibitors with the catalytic and diterpene sites of mACs. However, none of the AC inhibitors presently available have been comprehensively evaluated for isoform-specificity and -selectivity against other proteins. Targeting the diterpene site rather than the catalytic site may be a more promising strategy

Acknowledgments

We thank Drs Anshuman Dixit, Sara Dizayee, Michael B. Doughty, Stefan Dove, Michael Egger, Miriam Erdorf, Jens Geduhn, Martin Göttle, Jian-Xin Guo, Stefan Herzig, Klaus Höcherl, Roger A. Johnson, Melanie Hübner, Volkhard Kaever, Burkhard König, Prantik Maity, Jan Matthes, Cibele Pinto, Mark Richter, Dennis Rottländer, Michael Schäferling, Jennifer Schmidt, Yuequan Shen, Christian Spangler, Corinna Spangler, Philip Steindel, Srividya Suryanarayana, Hesham Taha, Stephen F. Vatner, Wei-Jen Tang,

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Trends in Pharmacological Sciences

ReviewInhibitors of membranous adenylyl cyclases

Section snippets

Membranous adenylyl cyclases (mACs)

Challenges to isoform-specific mAC inhibitors

Exploring the catalytic site of mACs with MANT- and TNP-nucleotides

Recent developments on P-site inhibitors

The diterpene site as target for mAC inhibitors

Remaining questions

Future directions

Concluding remarks

Acknowledgments

Trends Pharmacol. Sci.

J. Biol. Chem.

Trends Pharmacol. Sci.

Biochem. Pharmacol.

J. Biol. Chem.

J. Biol. Chem.

Comp. Biochem. Physiol. C

J. Biol. Chem.

Neurobiol. Dis.

J. Biol. Chem.

J. Biol. Chem.

Biochem. Pharmacol.

Methods Enzymol.

J. Biol. Chem.

Pharmacol. Ther.

Biochem. Pharmacol.

J. Biol. Chem.

J. Am. Coll. Cardiol.

J. Biol. Chem.

Pharmacol. Ther.

Mol. Cell. Endocrinol.

Genomics

J. Biol. Chem.

Trends Pharmacol. Sci.

Trends Pharmacol. Sci.

Biochem. Pharmacol.

J. Biol. Chem.

Brain Res.

J. Biol. Chem.

Neurosci. Lett.

Isoforms of mammalian adenylyl cyclase: multiplicities of signalling

Mol. Interv.

Physiological roles for G protein-regulated adenylyl cyclase isoforms: insights from knockout and overexpression studies

Neurosignals

Capturing adenylyl cyclases as potential drug targets

Nat. Rev. Drug Discov.

Identification of a forskolin-like molecule in human renal cysts

J. Am. Soc. Nephrol.

Modulation of β-adrenergic receptor signalling in heart failure and longevity targeting adenylyl cyclase type 5

Heart Fail. Rev.

Adenylyl cyclase isoforms as novel therapeutic targets: an exciting example of excitotoxicity neuroprotection

Mol. Interv.

Review
Inhibitors of membranous adenylyl cyclases