Adenosine analogue-oligo-arginine conjugates (ARCs) serve as high-affinity inhibitors and fluorescence probes of type I cGMP-dependent protein kinase (PKGIalpha).
BACKGROUND
Type I cGMP-dependent protein kinase (PKGIalpha) belongs to the family of cyclic nucleotide-dependent protein kinases and is one of the main effectors of cGMP. PKGIalpha is involved in regulation of cardiac contractility, vasorelaxation, and blood pressure; hence, the development of potent modulators of PKGIalpha would lead to advances in the treatment of a variety of cardiovascular diseases.


OBJECTIVE
Representatives of ARC-type compounds previously characterized as potent inhibitors and high-affinity fluorescent probes of PKA catalytic subunit (PKAc) were tested towards PKGIalpha to determine that ARCs could serve as activity regulators and sensors for the latter protein kinase both in vitro and in complex biological systems.


RESULTS
Structure-activity profiling of ARCs with PKGIalpha in vitro demonstrated both similarities as well as differences to corresponding profiling with PKAc, whereas ARC-903 and ARC-668 revealed low nanomolar displacement constants and inhibition IC(50) values with both cyclic nucleotide-dependent kinases. The ability of ARC-based fluorescent probes to penetrate cell plasma membrane was demonstrated in the smooth muscle tissue of rat cerebellum isolated arteries, and the compound with the highest affinity in vitro (ARC-903) showed also potential for in vivo applications, fully abolishing the PKG1alpha-induced vasodilation.
Lavogina
Darja
D
Institute of Chemistry, University of Tartu, Ravila 14a, 50411 Tartu, Estonia.
Nickl
Christian K
CK
Enkvist
Erki
E
Raidaru
Gerda
G
Lust
Marje
M
Vaasa
Angela
A
Uri
Asko
A
Dostmann
Wolfgang R
WR
D000241:Adenosine / Q000737:chemistry*; D000818:Animals; D001120:Arginine / Q000737:chemistry*; D002536:Cerebral Arteries / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D006152:Cyclic GMP / Q000378:metabolism; D062965:Cyclic GMP-Dependent Protein Kinase Type I; D017869:Cyclic GMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D005453:Fluorescence; D005456:Fluorescent Dyes; D006801:Humans; D009131:Muscle, Smooth, Vascular / Q000187:drug effects*; D051381:Rats; D014664:Vasodilation / Q000187:drug effects
D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; D001120:Arginine; D062965:Cyclic GMP-Dependent Protein Kinase Type I; D017869:Cyclic GMP-Dependent Protein Kinases; C569718:PRKG1 protein, human; D006152:Cyclic GMP; D000241:Adenosine
J Med Chem. 2006 Nov 30;49(24):7150-9
17125267
Arch Oral Biol. 2007 Oct;52(10):905-10
17559798
Handb Exp Pharmacol. 2009;(191):533-48
19089344
Anal Biochem. 2005 May 1;340(1):165-70
15802142
Annu Rev Pharmacol Toxicol. 1992;32:377-97
1605572
FEBS J. 2008 Jul;275(14):3608-24
18540886
Blood. 2004 Apr 1;103(7):2593-600
14644996
Adv Drug Deliv Rev. 2008 Mar 1;60(4-5):598-607
18045727
Anal Biochem. 2000 May 15;281(1):62-7
10847611
Front Biosci. 2005 May 01;10:1302-12
15769626
Biochim Biophys Acta. 2010 Mar;1804(3):524-32
20018259
J Biol Chem. 2009 Jun 12;284(24):16354-16368
19369251
Bioorg Med Chem Lett. 2007 Oct 1;17(19):5336-9
17716894
J Biol Chem. 1990 Mar 25;265(9):5267-72
2156866
Mol Pharmacol. 2004 May;65(5):1111-9
15102939
Biochem J. 2003 Apr 1;371(Pt 1):199-204
12534346
Res Commun Chem Pathol Pharmacol. 1991 Oct;74(1):3-14
1666198
Handb Exp Pharmacol. 2009;(191):137-62
19089329
Biochem J. 2009 May 13;420(2):179-89
19216719
Circ Res. 2006 Apr 28;98(8):993-1001
16645149
J Biol Chem. 2000 Oct 27;275(43):33536-41
10922374
Bioorg Med Chem Lett. 1999 May 17;9(10):1447-52
10360754
Anal Biochem. 2009 Feb 1;385(1):85-93
19017524
Proc Natl Acad Sci U S A. 2000 Oct 24;97(22):11996-2001
11050231
Physiol Rev. 2004 Jan;84(1):137-67
14715913
Cell Signal. 1997 May-Jun;9(3-4):269-76
9218127
Anal Biochem. 2007 Mar 15;362(2):268-77
17274940
J Biol Chem. 1996 Jan 5;271(1):174-9
8550555
Annu Rev Physiol. 2006;68:345-74
16460276
Peptides. 2009 Dec;30(12):2323-9
19698755
Biochim Biophys Acta. 2010 Mar;1804(3):541-6
19879386
Br J Pharmacol. 2008 Dec;155(7):984-94
18846041
Eur J Pharmacol. 1994 Oct 14;269(2):265-8
7851503
J Biol Chem. 2009 Oct 23;284(43):29260-8
19726669
J Control Release. 2009 Dec 16;140(3):237-44
19409429
Circ Res. 2007 Nov 26;101(11):1078-80
18040024
Bioconjug Chem. 2008 Dec;19(12):2535-42
19012426
Neuropsychopharmacology. 2009 Jun;34(7):1829-42
19194373
J Med Chem. 2009 Jan 22;52(2):308-21
19143565
Trends Endocrinol Metab. 2008 May-Jun;19(4):130-7
18337116
Am J Physiol Gastrointest Liver Physiol. 2003 Jun;284(6):G1006-16
12736149
Proc Natl Acad Sci U S A. 2000 Dec 19;97(26):14772-7
11121077
Biochemistry. 1984 Oct 9;23(21):5036-41
6238627
Bioorg Med Chem Lett. 2003 Sep 15;13(18):3035-9
12941328
J Biol Chem. 1997 Aug 8;272(32):20011-20
9242671
Alcohol Clin Exp Res. 2009 Apr;33(4):610-6
19183138
Pharmacol Ther. 1999 May-Jun;82(2-3):373-87
10454213
Bioorg Med Chem Lett. 2009 Nov 1;19(21):6098-101
19800227
Acta Pharmacol Sin. 2009 Oct;30(10):1399-414
19801996
Eur J Pharmacol. 2008 May 31;586(1-3):283-7
18420189
Circ Res. 2003 Nov 14;93(10):907-16
14615494
J Control Release. 2009 Oct 15;139(2):108-17
19577599
J Biol Chem. 2009 Dec 4;284(49):33957-65
19833724
ChemMedChem. 2010 Jan;5(1):23-34
19774589
Biochem J. 2002 Mar 15;362(Pt 3):533-7
11879178
Proc Natl Acad Sci U S A. 2001 Feb 27;98(5):2437-42
11226257
Pharmacol Ther. 2002 Feb-Mar;93(2-3):203-15
12191612
Physiology (Bethesda). 2009 Dec;24:342-56
19996365
Br J Pharmacol. 1995 Dec;116(8):3110-6
8719784
Gastroenterology. 2010 Jan;138(1):13-8.e1-3
19931260