Functional sites of chemically modified tetracyclines: inhibition of the oxidative activation of human neutrophil and chicken osteoclast pro-matrix metalloproteinases.
OBJECTIVE
We studied the relative ability of 6 different chemically modified non-antimicrobial analogs of tetracycline (CMT) to inhibit human and chicken matrix metalloproteinases (MMP) in vitro. The ability of tetracycline and its analogs to inhibit MMP appears to depend on the Ca++/Zn++ binding site at C11 (carbonyl oxygen) and C12 (OH group) of the molecule, which is lacking in CMT-5, the pyrazole derivative of tetracycline. This significant property of CMT-5 was used to differentiate between the effects of CMT on already active MMP versus the oxidative activation of latent MMP (pro-MMP).


METHODS
Cultured chicken osteoclast conditioned medium and purified human neutrophil progelatinase (MMP-9) and pro-collagenase (MMP-8) were assayed for proteinase activities using gelatin and collagen, respectively. The pro-MMP were activated either by preincubation with 1 mM aminophenylmercuric acetate (APMA) or 100 microM sodium hypochlorite (NaOCI). CMT were added either to the preincubation mixtures together with NaOCl or after activation of pro-MMP with NaOCl.


RESULTS
All CMT tested, except CMT-5, inhibited APMA or NaOCl activated pro-MMP. However, CMT-5 (like the other CMT), inhibited the oxidative activation of pro-MMP by NaOCl when added together by scavenging the reactive oxygen species. The degradation of type-I collagen by chicken osteoclast conditioned medium was probably due to MMP-2 and/or MMP-13.


CONCLUSIONS
Oxidative activation of pro-MMP may be crucial during soft tissue/bone destruction in the inflammatory diseases, including the arthritides. Our results indicate that the Ca++/Zn++ binding site of CMT is not essential for inhibition of the oxidative activation of pro-MMP.
Sorsa
T
T
Department of Medical Chemistry, Institute of Dentistry, University of Helsinki and Helsinki University Central Hospital, Finland.
Ramamurthy
N S
NS
Vernillo
A T
AT
Zhang
X
X
Konttinen
Y T
YT
Rifkin
B R
BR
Golub
L M
LM
D000818:Animals; D002645:Chickens; D017364:Collagenases / Q000378:metabolism; D004789:Enzyme Activation; D018093:Gelatinases / Q000378:metabolism; D006801:Humans; D006997:Hypochlorous Acid / Q000494:pharmacology; D020784:Matrix Metalloproteinase 8; D020780:Matrix Metalloproteinase 9; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D009504:Neutrophils / Q000187:drug effects* / Q000201:enzymology; D010010:Osteoclasts / Q000187:drug effects* / Q000201:enzymology; D013754:Tetracyclines / Q000737:chemistry / Q000494:pharmacology*
D013754:Tetracyclines; D006997:Hypochlorous Acid; D017364:Collagenases; D018093:Gelatinases; D008666:Metalloendopeptidases; D020784:Matrix Metalloproteinase 8; D020780:Matrix Metalloproteinase 9