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Minimal amidine structure for inhibition of nitric oxide biosynthesis

Minimal amidine structure for inhibition of nitric oxide biosynthesis
Minimal amidine structure for inhibition of nitric oxide biosynthesis
Pharmacological modulation of nitric oxide synthase activity has been achieved using structural analogs of arginine. In the present studies, we demonstrated that the minimal amidine structure required for enzymatic inhibition is formamidine. We found that the production of nitric oxide by primary cultures of rat hepatocytes and several mouse and human cell lines, including RAW 264.7 macrophages, PAM 212 keratinocytes, G8 myoblasts, S180 sarcoma, CX-1 human colon cells, and GH3 rat pituitary cells, was inhibited in a concentration- and time-dependent manner by formamidine. Formamidine was 2- to 6-fold more effective in inhibiting nitric oxide production in cells expressing inducible nitric oxide synthase (NOS2) than in a cell line expressing calcium-dependent neuronal nitric oxide synthase (NOS1). Whereas formamidine had no effect on gamma-interferon-induced expression of nitric oxide synthase protein, its enzymatic activity was blocked. Kinetic analysis revealed that formamidine acts as a simple competitive inhibitor with respect to arginine (K(i) formamidine approximately 800 microM). Using a polarographic microsensor to measure real-time flux of nitric oxide release from RAW 264.7 macrophages, formamidine was found to require 30-90 min to inhibit enzyme activity, suggesting that cellular uptake of the drug may limit its biological activity. Our data indicate that formamidine is an effective inhibitor of nitric oxide production. Furthermore, its low toxicity may make it useful as a potential therapeutic agent in diseases associated with the increased production of nitric oxide
0006-2952
1581-1586
Billack, Blase
ee08b534-cf8d-4d8d-a4ed-686d094ef52f
Heck, Diane E.
03a96ab5-efa2-4374-a29f-a9ec70101fc0
Porterfield, D. Marshall
ad367ffb-cd69-4c9e-ac0c-f7f048538518
Malchow, R .Paul
bbd9c97e-a519-4598-8687-ec7cbc57790c
Smith, Peter J.S.
003de469-9420-4f12-8f0e-8e8d76d28d6c
Gardner, Carol R.
c7686b6c-9b5d-4d95-8c23-b3b454f40bb7
Laskin, Debra L.
2944aba0-c866-4a7b-8463-fc612aa56a92
Laskin, Jeffrey D.
8c4377b4-ba21-4811-8cb1-35904f35e1d5
Billack, Blase
ee08b534-cf8d-4d8d-a4ed-686d094ef52f
Heck, Diane E.
03a96ab5-efa2-4374-a29f-a9ec70101fc0
Porterfield, D. Marshall
ad367ffb-cd69-4c9e-ac0c-f7f048538518
Malchow, R .Paul
bbd9c97e-a519-4598-8687-ec7cbc57790c
Smith, Peter J.S.
003de469-9420-4f12-8f0e-8e8d76d28d6c
Gardner, Carol R.
c7686b6c-9b5d-4d95-8c23-b3b454f40bb7
Laskin, Debra L.
2944aba0-c866-4a7b-8463-fc612aa56a92
Laskin, Jeffrey D.
8c4377b4-ba21-4811-8cb1-35904f35e1d5

Billack, Blase, Heck, Diane E., Porterfield, D. Marshall, Malchow, R .Paul, Smith, Peter J.S., Gardner, Carol R., Laskin, Debra L. and Laskin, Jeffrey D. (2001) Minimal amidine structure for inhibition of nitric oxide biosynthesis. Biochemical Pharmacology, 61 (12), 1581-1586. (doi:10.1016/S0006-2952(01)00630-X). (PMID:11377388)

Record type: Article

Abstract

Pharmacological modulation of nitric oxide synthase activity has been achieved using structural analogs of arginine. In the present studies, we demonstrated that the minimal amidine structure required for enzymatic inhibition is formamidine. We found that the production of nitric oxide by primary cultures of rat hepatocytes and several mouse and human cell lines, including RAW 264.7 macrophages, PAM 212 keratinocytes, G8 myoblasts, S180 sarcoma, CX-1 human colon cells, and GH3 rat pituitary cells, was inhibited in a concentration- and time-dependent manner by formamidine. Formamidine was 2- to 6-fold more effective in inhibiting nitric oxide production in cells expressing inducible nitric oxide synthase (NOS2) than in a cell line expressing calcium-dependent neuronal nitric oxide synthase (NOS1). Whereas formamidine had no effect on gamma-interferon-induced expression of nitric oxide synthase protein, its enzymatic activity was blocked. Kinetic analysis revealed that formamidine acts as a simple competitive inhibitor with respect to arginine (K(i) formamidine approximately 800 microM). Using a polarographic microsensor to measure real-time flux of nitric oxide release from RAW 264.7 macrophages, formamidine was found to require 30-90 min to inhibit enzyme activity, suggesting that cellular uptake of the drug may limit its biological activity. Our data indicate that formamidine is an effective inhibitor of nitric oxide production. Furthermore, its low toxicity may make it useful as a potential therapeutic agent in diseases associated with the increased production of nitric oxide

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Published date: June 2001
Organisations: University of Southampton

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Local EPrints ID: 188843
URI: https://eprints.soton.ac.uk/id/eprint/188843
ISSN: 0006-2952
PURE UUID: b0e155b3-5c8f-4bd5-86d8-ca88f76ccd0f
ORCID for Peter J.S. Smith: ORCID iD orcid.org/0000-0003-4400-6853

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Date deposited: 13 Jun 2011 12:45
Last modified: 06 Jun 2018 12:31

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Author: Blase Billack
Author: Diane E. Heck
Author: D. Marshall Porterfield
Author: R .Paul Malchow
Author: Carol R. Gardner
Author: Debra L. Laskin
Author: Jeffrey D. Laskin

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