Hydroxamic Group Research Articles

Pyran-containing sulfonamide hydroxamic acids: potent MMP inhibitors that spare MMP-1

The SAR of a series of sterically hindered sulfonamide hydroxamic acids with relatively large P1′ groups is described. The compounds typically spare MMP-1 while being potent inhibitors of MMP-13. The metabolically more stable compounds in the series contain either a monocyclic or bicyclic pyran ring adjacent to the hydroxamate group. Despite the sparing of MMP-1, pre-clinical and clinical studies revealed that fibrosis in rats and MSS in humans is still produced.

Transition metal complexes of acetamidomalondihydroxamate: synthesis, spectral, thermal and electrochemical studies

Acetamidomalondihydroxamate (K2AcAMDH) and its manganese(II), iron(II), cobalt(II), nickel(II), copper(II) and zinc(II) complexes were synthesized and characterized by elemental analysis, UV–VIS, IR and magnetic susceptibility. The pK a1 and pK a2 values of the dihydroxamic acid in aqueous solution were found to be 8.0 ± 0.1 and 9.7 ± 0.1. The dihydroxamate anion AcAMDH behaves as a tetradentate bridging ligand through both hydroxamate groups, forming complexes with a metal to ligand ratio of 1 : 1 in the solid state. The FTIR spectra and thermal decompositions of the ligand and its metal complexes were recorded. The redox behavior of the complexes was investigated in aqueous solution by square wave voltammetry and cyclic voltammetry at neutral pH. In contrast to the solid state, in solution the copper(II) and zinc(II) ions form stable complex species with a metal to ligand ratio of 1 : 2. The iron(II) and nickel(II) complexes show a two-electron irreversible reduction behavior, while the copper(II) and zinc(II) complexes undergo reversible electrode reactions. The stability constants of the complexes were determined by square wave voltammetry.

Gallium recovery from Bayer's liquor using hydroxamic acid resin

AbstractGallium, which is extensively used in the production of semiconductor materials, is present at the parts per million level in Bayer's liquor. The low concentration of gallium in the liquor, along with the high concentration of aluminum, prompted the use of chelating ion exchangers as an alternative separation process. A chelating ion exchange resin with hydroxamic groups attached to the copolymer of acrylonitrile‐divinylbenzene has been prepared by suspension polymerization followed by hydrolysis and chelation with hydroxylamine hydrochloride. Adsorption studies of gallium, using the above hydroxamic acid resin, were carried out. Adsorption was dependent on particle size of the resin and optimum conditions are determined for obtaining 0.3‐ to 0.5‐mm particles by varying the composition of the emulsion, using a secondary polymerization technique, and adding of diluents. Acrylic acid, as a diluent in the copolymer matrix, was found to increase the particle size and stability of the resin. IR studies, carried out for the products obtained at various stages, confirmed the conversion of polymer to resin with a hydroxamic acid group and its complex formation with gallium. Both batch and column studies were carried out for the determination of the capacity of the resin with synthetic Bayer's liquor containing gallium and commercial Bayer's liquor. Scaled‐up column studies were carried out with commercial Bayer's liquor to test the cyclability and stability of the resin. It was observed that chelated ion exchange resin could be recycled up to 30 times. The optimal liquid to solid phase ratio was found to be 1:12. © 2004 Wiley Periodicals, Inc. J Appl Polym Sci 92: 847–855, 2004

3-(4-Aroyl-1-methyl-1H-2-pyrrolyl)-N-hydroxy-2-propenamides as a new class of synthetic histone deacetylase inhibitors. 2. Effect of pyrrole-C2 and/or -C4 substitutions on biological activity.

Previous SAR studies (Part 1: Mai, A.; et al. J. Med. Chem. 2003, 46, 512-524) performed on some portions (pyrrole-C4, pyrrole-N1, and hydroxamate group) of 3-(4-benzoyl-1-methyl-1H-pyrrol-2-yl)-N-hydroxy-2-propenamide (1a) highlighted its 4-phenylacetyl (1b) and 4-cynnamoyl (1c) analogues as more potent compounds in inhibiting maize HD2 activity in vitro. In the present paper, we investigated the effect on anti-HD2 activity of chemical substitutions performed on the pyrrole-C2 ethene chains of 1a-c, which were replaced with methylene, ethylene, substituted ethene, and 1,3-butadiene chains (compounds 2). Biological results clearly indicated the unsubstituted ethene chain as the best structural motif to get the highest HDAC inhibitory activity, the sole exception to this rule being the introduction of the 1,3-butadienyl moiety into the 1a chemical structure (IC50(2f) = 0.77 microM; IC50(1a) = 3.8 microM). IC50 values of compounds 3, prepared as 1b homologues, revealed that between benzene and carbonyl groups at the pyrrole-C(4) position a hydrocarbon spacer length ranging from two to five methylenes is well accepted by the APHA template, being that 3a (two methylenes) and 3d (five methylenes) are more potent (2.3- and 1.4-fold, respectively) than 1b, while the introduction of a higher number of methylene units (see 3e,f) decreased the inhibitory activities of the derivatives. Particularly, 3a (IC50 = 0.043 microM) showed the same potency as SAHA in inhibiting HD2 in vitro, and it was 3000- and 2.6-fold more potent than sodium valproate and HC-toxin and was 4.3- and 6-fold less potent than trapoxin and TSA, respectively. Finally, conformationally constrained forms of 1b,c (compounds 4), prepared with the aim to obtain some information potentially useful for a future 3D-QSAR study, showed the same (4a,b) or higher (4c,d) HD2 inhibiting activities in comparison with those of the reference drugs. Molecular modeling and docking calculations on the designed compounds performed in parallel with the chemistry work fully supported the synthetic effort and gave insights into the binding mode of the more flexible APHA derivatives (i.e., 3a). Despite the difference of potency between 1b and 3a in the enzyme assay, the two APHA derivatives showed similar antiproliferative and cytodifferentiating activities in vivo on Friends MEL cells, being that 3a is more potent than 1b in the differentiation assay only at the highest tested dose (48 microM).

Bacterial iron transport: coordination properties of azotobactin, the highly fluorescent siderophore of Azotobacter vinelandii.

Azotobacter vinelandii, a nitrogen-fixing soil bacterium, secretes in iron deficiency azotobactin delta, a highly fluorescent pyoverdin-like chromopeptidic hexadentate siderophore. The chromophore, derived from 2,3-diamino-6,7 dihydroxyquinoline, is bound to a peptide chain of 10 amino acids: (L)-Asp-(D)-Ser-(L)-Hse-Gly-(D)-beta-threo-HOAsp-(L)-Ser-(D)-Cit-(L)-Hse-(L)-Hse lactone-(D)-N(delta)-Acetyl, N(delta)-HOOrn. Azotobactin delta has three different iron(III) binding sites which are one hydroxamate group at the C-terminal end of the peptidic chain (N(delta)-Acetyl, N(delta)-HOOrn), one alpha-hydroxycarboxylic function in the middle of the chain (beta-threo-hydroxyaspartic acid), and one catechol group on the chromophore. The coordination properties of its iron(III) and iron(II) complexes were measured by spectrophotometry, potentiometry, and voltammetry after the determination of the acid-base functions of the uncomplexed free siderophore. Strongly negatively charged ferric species were observed at neutral p[H]'s corresponding to a predominant absolute configuration Lambda of the ferric complex in solution as deduced from CD measurements. The presence of an alpha-hydroxycarboxylic chelating group does not decrease the stability of the iron(III) complex when compared to the main trishydroxamate siderophores or to pyoverdins. The value of the redox potential of ferric azotobactin is highly consistent with a reductive step by physiological reductants for the iron release. Formation and dissociation kinetics of the azotobactin delta ferric complex point out that both ends of this long siderophore chain get coordinated to Fe(III) before the middle. The most striking result provided by fluorescence measurements is the lasting quenching of the fluorophore in the course of the protonation of the ferric azotobactin delta complex. Despite the release of the hydroxyacid and of the catechol, the fluorescence remains indeed quenched, when iron(III) is bound only to the hydroxamic acid, suggesting a folded conformation at this stage, around the metal ion, in contrast to the unfolded species observed for other siderophores such as ferrioxamine or pyoverdin PaA.

C–N bond formation followed by N–Cl bond breaking. One more and unexpected case of the formation of a hydroxamic group via heterolytic bond cleavage

An unexpected and previously unknown reaction sequence in the interactions of the acyl halides with nitrosobenzenes, which involves carbon–nitrogen bond formation followed by heterolytic nitrogen–chlorine bond cleavage giving the corresponding unsubstituted N-phenylalkylhydroxamic acids (or N-phenylarylhydroxamic acids) and chlorine as the products has been observed. The kinetic and other evidence obtained suggest that the carbon–nitrogen bond formation is the consequence of a nucleophilic interaction of an N-phenylchlorohydroxylamine intermediate, formed in the first reaction step, with the acyl halide in the second step of the complex sequence, which leads to an N-acyl- N-chlorophenylhydroxylamine cation intermediate. The key reaction step involves the interaction of an N-acyl- N-chlorophenylhydroxylamine cation intermediate with chloride ion, which leads to the N–Cl heterolytic bond cleavage and the final formation of the hydroxamic group and a molecule of chlorine.

Hydroxamic acids: proton donor and acceptor strength for use in drug design

Hydroxamic acids, the naturally occurring and synthetic products, generally have low toxicities and are of interest for many therapeutic applications. The present investigation describes the measurement of hydrogen bond donor (HBD) strength of ten hydroxamic acids by measuring their log P(O/W) values. Hydroxamic acid functional group contains two oxygen and one nitrogen atom as the acceptor sites. Thus, HBA strength of these reagents is also computed. A knowledge of these parameters is valuable in the field of toxicology, pharmacology and environmental sciences.

Synthesis of conducting poly[di-heteroaromatics] from pyrrole, indole, and carbazole, and their mixed hydroxamates, and studies of their metal complexes

Compounds 1,1-di-(N-pyrrolyl)methane hydroxamic acid (II), 1,1-di-(N-indolyl)methane hydroxamic acid (IV), 1,1-di-(N-carbazolyl)methane hydroxamic acid (VI), 1-(N-pyrrolyl)-1-(N-carbazolyl)methane hydroxamic acid (VIII), and 1-(N-pyrrolyl)-1-(N-indolyl)-methane hydroxamic acid (X) were synthesized from their N,N-substituted ester group derivatives. They were chemically polymerized in the presence of oxidants such as ammonium persulfate and iron(III) perchlorate for hydroxamic acid and ester groups derivatives, respectively. The conductivity of these polymers was measured by a four-point probe method. The complexes of hydroxamic acid derivatives with certain ions [such as Fe(III), Rh(III), Ir(III), Cr(III), Cu(II), and Ni(II)] were studied. Compounds II, IV, and VI and their ester group derivatives were successfully electropolymerized by a cyclic voltammetry method under the following conditions: 6 × 10−3 M of monomer, 0.1 M lithium perchlorate, 20 mL of acetonitrile, glassy carbon (GC) disk working electrode, and Ag/AgCl reference electrode and platinum wire counter electrode conditions. © 2003 Wiley Periodicals, Inc. J Appl Polym Sci 90: 63–71, 2003

Syntheses, Spectral, Thermal and Electrochemical Studies of 3-Carboxylacetonehydroxamic Acid and its Iron(Ii), Cobalt(Ii), Nickel(Ii), Copper(Ii) and Zinc(Ii) Complexes

3-Carboxylacetonehydroxamic acid (CAHA) and its iron(II), cobalt(II), nickel(II), copper(II) and zinc(II) complexes were synthesized and characterized by elemental analysis, UV-Vis and IR spectra and magnetic susceptibility. The pK a1 and pK a2 values of the ligand in aqueous solution were found to be 6.5 ± 0.1 and 8.6 ± 0.1, which correspond to dissociation of carboxyl and hydroxamic protons, respectively. The dianion CAH acts as a tetradentate ligand through the hydroxamate and carboxylate groups and coordinates to the divalent metal ions, forming coordination polymers with a metal-to-ligand ratio of 1 : 1 in the solid state. FTIR spectra and thermal decomposition of the ligand and its metal complexes were recorded and briefly discussed. The electrochemical behavior of the complexes was investigated by square wave voltammetry and cyclic voltammetry at neutral pH. In contrast to the solid state, the iron(II) and copper(II) cations form stable complex species with a metal-to-ligand ratio of 1 : 2 in solution. The iron(II), cobalt(II) and nickel(II) complexes show two-electron irreversible reduction behavior, while the copper(II) and zinc(II) complexes undergo quasi-reversible and reversible electrode reactions, respectively. The stability constants of the complexes were determined by square wave voltammetry.

Iron chelation properties of an extracellular siderophore exochelin MN.

The coordination chemistry of an extracellular siderophore produced by Mycobacterium neoaurum, exochelin MN (ExoMN), is reported along with its pK(a) values, Fe(III) and Fe(II) chelation constants, and aqueous solution speciation as determined by spectrophotometric and potentiometric titration techniques. Exochelin MN is of particular interest as it can efficiently transport iron into pathogenic M. leprae, which is responsible for leprosy, in addition to its own parent cells. The Fe(III) coordination properties of ExoMN are important with respect to understanding the Fe(III) acquisition and uptake mechanism in pathogenic M. leprae, as the siderophores from this organism are very difficult to isolate. Exochelin MN has two hydroxamic acid groups and an unusual threo-beta-hydroxy-l-histidine available for Fe(III) chelation. The presence of threo-beta-hydroxy-l-histidine gives rise to a unique mode of Fe(III) coordination. The pK(a) values for the two hydroxamic acid moieties, the histidine imidazole ring and the alkylammonium groups on ExoMN, correspond well with the literature values for these moieties. Proton-dependent Fe(III)- and Fe(II)-ExoMN equilibrium constants were determined using a model involving sequential protonation of the Fe(III)- and Fe(II)-ExoMN complexes. These data were used to develop a model whereby deprotonation reactions on the surface of the complex in the second coordination shell result in first coordination shell isomerization. The overall formation constants were calculated: log beta(110) = 39.12 for Fe(III)-ExoMN and 16.7 for Fe(II)-ExoMN. The calculated pFe value of 31.1 is one of the highest among all siderophores and their synthetic analogues and indicates that ExoMN is thermodynamically capable of removing Fe(III) from transferrin. The E(1/2) for the Fe(III)ExoMN/Fe(II)ExoMN(-) couple was determined to be -595 mV from quasi-reversible cyclic voltammograms at pH = 10.8, and the pH-dependent E(1/2) profile was used to determine the Fe(II)-ExoMN protonation constants.

Synthesis and biological evaluation of hydroxamate-Based inhibitors of glutamate carboxypeptidase II.

A series of hydroxamic acids has been prepared as potential inhibitors of glutamate carboxypeptidase II (GCP II). Compounds based on a P1' residue (primed-side inhibitors) were more potent than those based on a P1 group (unprimed-side inhibitors). Inhibitory potency of the primed-side GCP II inhibitors was found to be dependent on the number of methylene units between the hydroxamate group and pentanedioic acid. Succinyl hydroxamic acid derivative, 2-(hydroxycarbamoylmethyl)pentanedioic acid, is the most potent GCP II inhibitor with an IC(50) value of 220nM. The comparison of the results to those of other classes of GCP II inhibitors as well as hydroxamate-based MMP inhibitors provides further insight into the structure-activity relationships of GCP II inhibition.

Intramolecular Monodealkylation During the Attempted Synthesis of Diethylphosphonoacetohydroxamic Acid.

AbstractFor Abstract see ChemInform Abstract in Full Text.

Photochemical reactivity of siderophores produced by marine heterotrophic bacteria and cyanobacteria based on characteristic Fe(III) binding groups

Siderophores, high‐affinity Fe(III) ligands produced by microorganisms to facilitate iron acquisition, might contribute significantly to dissolved Fe(III) complexation in ocean surface waters. In previous work, we demonstrated the photoreactivity of the ferric ion complexes of several α‐hydroxy carboxylic acid—containing siderophores produced by heterotrophic marine bacteria. Here, we expand on our earlier studies and detail the photoreactivity of additional siderophores produced by both heterotrophic marine bacteria and marine cyanobacteria, making comparisons to synthetic and terrestrial siderophores that lack the α‐hydroxy carboxylate group. Our results suggest that, in addition to secondary photochemical reaction pathways involving reactive oxygen species, direct photolysis of Fe(III)‐siderophore complexes might be a significant source of Fe(II) and reactive Fe(III) in ocean surface waters. Our findings further indicate that the photoreactivity of siderophores is primarily determined by the chemical structure of the Fe(III) binding groups that they possess—hydroxamate, catecholate, or α‐hydroxy carboxylate moieties. Hydroxamate groups are photochemically resistant regardless of Fe(III) complexation. Catecholates, in contrast, are susceptible to photooxidation in the uncomplexed form but stabilized against photooxidation when ferrated. α‐Hydroxy carboxylate groups are stable as the uncomplexed acid, but when coordinated to Fe(III), these moieties undergo light‐induced ligand oxidation and reduction of Fe(III) to Fe(II). These photochemical properties appear to determine the reactivity and fate of Fe(III)‐binding siderophores in ocean surface waters, which in turn might significantly influence the biogeochemical cycling of iron.

Metal coordination-based inhibitors of adenylyl cyclase: novel potent P-site antagonists.

The adenylyl cyclases (ACs) are a family of intracellular enzymes associated with signal transduction by virtue of their ability to convert ATP to cAMP. The catalytic mechanism of this transformation proceeds through initial binding of ATP to the so-called purine binding site (P-site) of the enzyme followed by metal-mediated cyclization with loss of pyrophosphate. Crystallographic analysis of ACs with known inhibitors reveals the presence of two metals in the active site. Presently, nine isoforms of adenylyl cyclase are known, and unique isoform combinations are expressed in a tissue-specific manner. The development of isoform-specific inhibitors of adenylyl cyclase may prove to be a useful strategy toward the design of unique signal transduction inhibitors. To develop novel AC inhibitors, we have chosen an approach to inhibitor design utilizing an adenine ring system joined to a metal-coordinating hydroxamic acid via various linkers. Previous work in our group has validated this approach and identified novel inhibitors that possess an adenine ring joined to a metal-coordinating hydroxamic acid through flexible acyclic linkers (Levy, D. E., et al. Bioorg. Med. Chem. Lett. 2002, 12, 3085-3088). Subsequent studies have focused on the introduction of conformational restrictions into the tether of the inhibitors with the goal of increasing potency (Levy, D. E., et al. Bioorg. Med. Chem. Lett. 2002, 12, 3089-3092). Building upon the favorable spatial positioning of the adenine and hydroxamate groups coupled with potentially favorable entropic factors, the unit joining the carbocycle to the hydroxamate was explored further and a stereochemical-based SAR was elucidated, leading to a new series of highly potent AC inhibitors.

Synthesis of new chelating ion exchange resin with hydroxamic and amidoxime groups and study of its metal binding property

AbstractThe synthesis of a new chelating ion exchanger with hydroxamic and amidoxime groups was performed by an aminolysis reaction of an ethylacrylate/acrylonitrile/divinylbenzene copolymer with hydroxylamine in the presence of sodium ethoxylate. We studied the retention property of the new resin for Pb(II), Cd(II), Cu(II), Ni(II), Zn(II), Mn(II), and Fe(III) metal cations from aqueous solutions. The retention capacity depends on the contact time, the pH of the solution, and the types of counterions of the metal cations. Competitive adsorptions of Pb(NO3)2 + Fe2(SO4)3, Pb(NO3)2 + MnSO4, Pb(NO3)2 + Cd(NO3)2, and Pb(NO3)2 + CuCl2 on the new chelating ion exchanger were performed. The results showed that the resin preferentially retained the Pb(II) ions from the binary aqueous mixtures. © 2003 Wiley Periodicals, Inc. J Appl Polym Sci 88: 2956–2962, 2003

Preferential extraction of gallium from Bayer liquor using ion exchange chelating resin containing hydroxamic acid functional group

AbstractGallium extraction studies were carried out using resin containing an hydroxamic acid functional group. Extraction was dependant on reaction time and alkali concentration. The resin did not extract aluminium. Vanadium extraction kinetics were slower than those of gallium. The gallium and vanadium separation efficiency increased with decreasing contact time with extractions of 64% and 32% respectively after 1 min. Gallium extraction was independent of alkali concentration up to 250 g dm−3 and thereafter decreased with increasing alkali concentration. Vanadium extraction decreased with increasing alkali concentration. Cyclic batch and column studies were carried out with synthetic as well as actual Bayer liquor to test the stability and preferential extraction of gallium using the resin. It was observed that using resin partially loaded with gallium, the co‐extraction of vanadium could be minimised. Copyright © 2003 Society of Chemical Industry

A convenient parallel synthesis of low molecular weight hydroxamic acids using polymer-supported 1-hydroxybenzotriazole.

A convenient two-step procedure for the parallel synthesis of hydroxamic acids from carboxylic acids and hydroxylamine in good to high yields is reported. It involves the formation of a polymer-bound HOBt active ester and subsequent reaction with O-protected or free hydroxylamine. The hydroxamates are isolated with high purities by simple evaporation of volatile solvents. The use of free hydroxylamine leads to increased yields while maintaining high purities. Recycling of the spent resin to produce the same or a different hydroxamic acid has been achieved by a three-step protocol which is easily amenable to automation and cost-economical. The method presented here is well suited to the preparation of the title compounds and can be used effectively to synthesise large molecules containing a hydroxamic acid group.

Peptide-bond modification for metal coordination: peptides containing two hydroxamate groups.

Peptide-bond modification via N-hydroxylation has been explored as a strategy for metal coordination to induce conformational rigidity and orient side chains for specific molecular recognition. N-Hydroxyamides were prepared by reacting N-benzyloxyamino acid esters or amides with Fmoc-AA-Cl/AgCN (Fmoc: 9-fluorenylmethoxycarbonyl; AA: amino acid) in toluene or Fmoc-AA/HATU/DIEA in DMF (HATU: O-(7-azabenzotriazol-lyl)-1,1,3,3-tetramethyluronium hexafluorophosphate; DIEA: N,N-diisopropylethylamine; DMF: N,N-dimethylformamide), followed by deblocking of benzyl protecting groups. Novel linear and cyclic N,N'-dihydroxypeptides were efficiently assembled using Fmoc chemistry in solution and/or on a solid support. As screened by electrospray ionization-mass spectroscopy (ESI-MS), high iron-binding selectivity and affinity were attainable. Compounds having a spacer of two alpha-amino acids between the amino acids bearing the two hydroxamates, i.e., a spacer of 8 atoms, generated 1:1 iron complex species in the gas phase. Moreover, high performance liquid chromatography (HPLC), uv/vis, and (1)H-NMR analyses provided direct evidence for complex formations in solution. Significantly, the representative compound cyclo(Leu-Psi[CON(OH)]-Phe-Ala-Pro)(2) (P8) may serve as a robust metal-binding scaffold in construction of a metal-binding library for versatile metal-mediated molecular recognition.

Intramolecular monodealkylation during the attempted synthesis of diethylphosphonoacetohydroxamic acid

AbstractThe reaction of diethyl methyl phosphonoacetate (1) with hydroxylamine in NaOH solution resulted in the loss of one of the phosphorus ethyl groups, and yielded monoethylphosphonoacetohydroxamic acid (2) as the major product (79%) and diethylphosphonoacetic acid (3) as the minor product (21%). A series of control experiments were carried out to elucidate the sequence of the reactions leading to 2. When the reaction of 1 with NH2OH was carried out in NaHCO3 solution, a transient product 4 was also observed, which slowly transformed to 2. Compound 4 was assigned the structure diethylphosphonoacetohydroxamic acid. There was no dealkylation observed at the phosphorus when 1 was reacted with methoxylamine or when O‐methyl diethylphosphonoacetohydroxamate (7) was placed in alkaline solution. The dealkylation at phosphorus was interpreted in terms of intramolecular nucleophilic catalysis by the hydroxamic OH group attacking the phosphorus in 4, involving cyclic 1,2,5‐oxazaphospholidine intermediates. © 2003 Wiley Periodicals, Inc. Heteroatom Chem 14:67–71, 2003; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/hc.10082

Rational design of tropoelastin peptide-based inhibitors of metalloproteinases.

Abnormal production of matrix metalloproteinases (MMPs) has been observed in a variety of diseases, such as emphysema, atherosclerosis, and cancer metastasis. Destruction of connective tissue ensues and elastin is often a key target. Three of the main elastolytic MMPs are the gelatinases MMP-2 and MMP-9 and the metalloelastase MMP-12. To investigate the possibility of using peptides to inhibit the elastolytic activity of these enzymes, we mapped the sites within tropoelastin recognized by MMP-9 and MMP-12. Peptides that correspond to regions overlapping these sites were then tested for their ability to inhibit these MMPs. These included an unmodified peptide directed against MMP-9 (peptide PP), cysteine-containing peptides that mimicked either the MMP-9 (peptide NCP) or the MMP-12 (peptide lin24) cleavage sites in tropoelastin and their cyclized forms (CP and cyc24, respectively), and a peptide containing a zinc-chelating hydroxamate group directed against MMP-9 (HP). The presence of a free sulfhydryl or hydroxamate group capable of chelating the zinc ion in the active site of the MMPs was generally found to increase the inhibitory activity of the peptides. The specificity of the inhibitors varied, with some of the inhibitors showing activity against all of the MMPs examined. None of the inhibitors had any significant effect on the activity of the unrelated serine protease, plasmin. K(i) values for the inhibitors were in the micromolar range. Our results suggest ways of developing other MMP inhibitors based on substrate recognition sites that may provide greater levels of inhibition.