Daniel F. Ortwine

Histone deacetylases (HDACs) are critical in the control of gene expression, and dysregulation of their activity has been implicated in a broad range of diseases, including cancer, cardiovascular, and neurological diseases. HDAC inhibitors (HDACi) employing different zinc chelating functionalities such as hydroxamic acids and benzamides have shown promising results in cancer therapy. Although it has also been suggested that HDACi with increased isozyme selectivity and potency may broaden their clinical utility and minimize side effects, the translation of this idea to the clinic remains to be investigated. Moreover, a detailed understanding of how HDACi with different pharmacological properties affect biological functions in vitro and in vivo is still missing. Here, we show that a panel of benzamide-containing HDACi are slow tight-binding inhibitors with long residence times unlike the hydroxamate-containing HDACi vorinostat and trichostatin-A. Characterization of changes in H2BK5 and H4K14 acetylation following HDACi treatment in the neuroblastoma cell line SH-SY5Y revealed that the timing and magnitude of histone acetylation mirrored both the association and dissociation kinetic rates of the inhibitors. In contrast, cell viability and microarray gene expression analysis indicated that cell death induction and changes in transcriptional regulation do not correlate with the dissociation kinetic rates of the HDACi. Therefore, our study suggests that determining how the selective and kinetic inhibition properties of HDACi affect cell function will help to evaluate their therapeutic utility.

Voltage-gated sodium (Nav) channels propagate action potentials in excitable cells. Accordingly, Nav channels are therapeutic targets for many cardiovascular and neurological disorders. Selective inhibitors have been challenging to design because the nine mammalian Nav channel isoforms share high sequence identity and remain recalcitrant to high-resolution structural studies. Targeting the human Nav1.7 channel involved in pain perception, we present a protein-engineering strategy that has allowed us to determine crystal structures of a novel receptor site in complex with isoform-selective antagonists. GX-936 and related inhibitors bind to the activated state of voltage-sensor domain IV (VSD4), where their anionic aryl sulfonamide warhead engages the fourth arginine gating charge on the S4 helix. By opposing VSD4 deactivation, these compounds inhibit Nav1.7 through a voltage-sensor trapping mechanism, likely by stabilizing inactivated states of the channel. Residues from the S2 and S3 helices are key determinants of isoform selectivity, and bound phospholipids implicate the membrane as a modulator of channel function and pharmacology. Our results help to elucidate the molecular basis of voltage sensing and establish structural blueprints to design selective Nav channel antagonists.

Bruton's tyrosine kinase (Btk) is a nonreceptor cytoplasmic tyrosine kinase involved in B-cell and myeloid cell activation, downstream of B-cell and Fcγ receptors, respectively. Preclinical studies have indicated that inhibition of Btk activity might offer a potential therapy in autoimmune diseases such as rheumatoid arthritis and systemic lupus erythematosus. Here we disclose the discovery and preclinical characterization of a potent, selective, and noncovalent Btk inhibitor currently in clinical development. GDC-0853 (29) suppresses B cell- and myeloid cell-mediated components of disease and demonstrates dose-dependent activity in an in vivo rat model of inflammatory arthritis. It demonstrates highly favorable safety, pharmacokinetic (PK), and pharmacodynamic (PD) profiles in preclinical and Phase 2 studies ongoing in patients with rheumatoid arthritis, lupus, and chronic spontaneous urticaria. On the basis of its potency, selectivity, long target residence time, and noncovalent mode of inhibition, 29 has the potential to be a best-in-class Btk inhibitor for a wide range of immunological indications.

Matrix metalloproteinase-13 (MMP13) is a Zn2+-dependent protease that catalyzes the cleavage of type II collagen, the main structural protein in articular cartilage. Excess MMP13 activity causes cartilage degradation in osteoarthritis, making this protease an attractive therapeutic target. However, clinically tested MMP inhibitors have been associated with a painful, joint-stiffening musculoskeletal side effect that may be due to their lack of selectivity. In our efforts to develop a disease-modifying osteoarthritis drug, we have discovered MMP13 inhibitors that differ greatly from previous MMP inhibitors; they do not bind to the catalytic zinc ion, they are noncompetitive with respect to substrate binding, and they show extreme selectivity for inhibiting MMP13. By structure-based drug design, we generated an orally active MMP13 inhibitor that effectively reduces cartilage damage in vivo and does not induce joint fibroplasias in a rat model of musculoskeletal syndrome side effects. Thus, highly selective inhibition of MMP13 in patients may overcome the major safety and efficacy challenges that have limited previously tested non-selective MMP inhibitors. MMP13 inhibitors such as the ones described here will help further define the role of this protease in arthritis and other diseases and may soon lead to drugs that safely halt cartilage damage in patients. Matrix metalloproteinase-13 (MMP13) is a Zn2+-dependent protease that catalyzes the cleavage of type II collagen, the main structural protein in articular cartilage. Excess MMP13 activity causes cartilage degradation in osteoarthritis, making this protease an attractive therapeutic target. However, clinically tested MMP inhibitors have been associated with a painful, joint-stiffening musculoskeletal side effect that may be due to their lack of selectivity. In our efforts to develop a disease-modifying osteoarthritis drug, we have discovered MMP13 inhibitors that differ greatly from previous MMP inhibitors; they do not bind to the catalytic zinc ion, they are noncompetitive with respect to substrate binding, and they show extreme selectivity for inhibiting MMP13. By structure-based drug design, we generated an orally active MMP13 inhibitor that effectively reduces cartilage damage in vivo and does not induce joint fibroplasias in a rat model of musculoskeletal syndrome side effects. Thus, highly selective inhibition of MMP13 in patients may overcome the major safety and efficacy challenges that have limited previously tested non-selective MMP inhibitors. MMP13 inhibitors such as the ones described here will help further define the role of this protease in arthritis and other diseases and may soon lead to drugs that safely halt cartilage damage in patients. The National Institutes of Health has estimated that more than 20 million adults in the United States suffer from osteoarthritis (OA), 3The abbreviations used are: OAosteoarthritisMMPmatrix metalloproteaseMSSmusculoskeletal syndromeCDcatalytic domainFLfull-lengthAUC0–24 harea under the curve from time 0 to 24 h.3The abbreviations used are: OAosteoarthritisMMPmatrix metalloproteaseMSSmusculoskeletal syndromeCDcatalytic domainFLfull-lengthAUC0–24 harea under the curve from time 0 to 24 h. a debilitating disease in which the protective cushion of cartilage is destroyed, resulting in pain and reduced mobility. A critical step in OA pathology is breakdown of the main structural protein of articular cartilage, type II collagen. This triple helical protein is resistant to most proteases but is efficiently recognized and degraded by the Zn2+-dependent enzyme, collagenase-3, known as matrix metalloproteinase-13 (MMP13) (1Massova L. Kotra P. Fridman R. Mobashery S. FASEB J. 1998; 12: 1075-1095Crossref PubMed Scopus (700) Google Scholar, 2Knäuper V. López-Otin C. Smith B. Knight G. Murphy G. J. Biol. Chem. 1996; 271: 1544-1550Abstract Full Text Full Text PDF PubMed Scopus (784) Google Scholar, 3Welgus H.G. Kobayashi D.K. Jeffrey J.J. J. Biol. Chem. 1983; 258: 14152-14165Google Scholar). MMP13 catalyzes the hydrolysis of type II collagen at a unique site resulting in ¾- and ¼-length polypeptide products (2Knäuper V. López-Otin C. Smith B. Knight G. Murphy G. J. Biol. Chem. 1996; 271: 1544-1550Abstract Full Text Full Text PDF PubMed Scopus (784) Google Scholar, 3Welgus H.G. Kobayashi D.K. Jeffrey J.J. J. Biol. Chem. 1983; 258: 14152-14165Google Scholar, 4Mitchell P.G. Magna H.A. Reeves L.M. Lopresti-Morrow L.L. Yocum S.A. Rosner P.J. Geoghegan K.F. Hambor J.E. J. Clin. Investig. 1996; 97: 761-768Crossref PubMed Scopus (827) Google Scholar, 5Billinghurst R.C. Dahlberg L. Ionescu M. Reiner A. Bourne R. Rorabeck C. Mitchell P. Hambor J. Diekmann O. Tschesche H. Chen J. Wart Van J. Clin. Investig. 1997; 99: 1534-1545Crossref PubMed Scopus (863) Google Scholar, 6Reboul P. Pelletier J J. Clin. Investig. 1996; 97: 2011-2019Crossref PubMed Scopus (425) Google Scholar). MMP13 is not found in normal adult tissues but is expressed in the joints and articular cartilage of OA patients (4Mitchell P.G. Magna H.A. Reeves L.M. Lopresti-Morrow L.L. Yocum S.A. Rosner P.J. Geoghegan K.F. Hambor J.E. J. Clin. Investig. 1996; 97: 761-768Crossref PubMed Scopus (827) Google Scholar, 5Billinghurst R.C. Dahlberg L. Ionescu M. Reiner A. Bourne R. Rorabeck C. Mitchell P. Hambor J. Diekmann O. Tschesche H. Chen J. Wart Van J. Clin. Investig. 1997; 99: 1534-1545Crossref PubMed Scopus (863) Google Scholar, 6Reboul P. Pelletier J J. Clin. Investig. 1996; 97: 2011-2019Crossref PubMed Scopus (425) Google Scholar, 7Wernicke D. Seyfert C. Hinzmann B. Gromnica-Ihle E. J. Rheumatol. 1996; 23: 590-595PubMed Google Scholar, 8Freemont A.J. Byers R.J. Taiwo Y.O. Hoyland J.A. Ann. Rheum. Dis. 1999; 58: 357-365Crossref PubMed Scopus (64) Google Scholar). In addition, regulated expression of human MMP13 in hyaline and joint cartilages induces OA in genetically modified mice (9Neuhold L.A. Killar L. Zhao W. Sung M J. Clin. Investig. 2001; 107: 35-44Crossref PubMed Scopus (440) Google Scholar). Furthermore, a MMP inhibitor that preferentially inhibits MMP13 has been shown to block the degradation of explanted human osteoarthritic cartilage (5Billinghurst R.C. Dahlberg L. Ionescu M. Reiner A. Bourne R. Rorabeck C. Mitchell P. Hambor J. Diekmann O. Tschesche H. Chen J. Wart Van J. Clin. Investig. 1997; 99: 1534-1545Crossref PubMed Scopus (863) Google Scholar). Based on these findings, it is likely that MMP13 is the direct cause of irreversible cartilage damage in OA. osteoarthritis matrix metalloprotease musculoskeletal syndrome catalytic domain full-length area under the curve from time 0 to 24 h. osteoarthritis matrix metalloprotease musculoskeletal syndrome catalytic domain full-length area under the curve from time 0 to 24 h. The clinical development of drugs that inhibit the actions of MMPs has been plagued by the association of a painful, joint-stiffening tendonitis-like side effect, termed “musculoskeletal syndrome” (MSS), with these inhibitors (10Clark I.M. Parker A.E. Expert Opin. Ther. Targets. 2003; 7: 19-34Crossref PubMed Scopus (75) Google Scholar, 11Drummond A.H. Beckett P. Brown P.D. Bone E.A. Davidson A.H. Galloway W.A. Gearing A.J.H. Huxley P. Laber D. McCourt M. Whittaker M. Wood L.M. Wright A. Ann. N. Y. Acad. Sci. 1999; 878: 228-235Crossref PubMed Scopus (198) Google Scholar). Such joint side effects are not unique to humans. Rats dosed with non-selective MMP inhibitors (i.e. compounds that inhibit several or all MMPs) also display MSS-like side effects such as soft tissue fibroplasias, inflammation, and pain (12Renkiewicz R. Qiu L. Lesch C. Sun X. Devalaraja R. Cody T. Kaldjian E. Welgus H. Baragi V. Arthritis Rheum. 2003; 48: 1742-1749Crossref PubMed Scopus (167) Google Scholar). Although the human joint side effects are reversible upon withdrawal of drug, MSS has halted clinical trials of many non-selective MMP inhibitors (10Clark I.M. Parker A.E. Expert Opin. Ther. Targets. 2003; 7: 19-34Crossref PubMed Scopus (75) Google Scholar). We began a search for MMP13-selective inhibitors with the hypothesis that they would effectively prevent cartilage degradation without causing MSS-like side effects. MMP Activity Assays-MMP13 activity and MMP selectivity assays employed a thioester substrate, acetyl-Pro-Leu-Gly-[2-mercapto-4-methyl-pentanoyl]-Leu-Gly-O-ethyl ester (Bachem Corp. #H-7145) (13Weingarten H. Feder J. Anal. Biochem. 1985; 147: 437-440Crossref PubMed Scopus (90) Google Scholar) and reaction mixtures containing 50 mm N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid (Hepes) buffer (pH 7.0), 10 mm CaCl2, 1 mm 5,5′-dithiobis(2-nitrobenzoic acid), 0.1 mm substrate, 0.005% Brij 35, and test compounds dissolved in Me2SO (1% final). A recombinant human MMP13 catalytic domain (CD) construct encompassing Tyr-104 to Asp-270 (SWISS-PROT P45452) was generated by synthetic gene methods and bacterial expression (14Biochem. Biophys. Res. Commun. 1992; 186: 143-149Crossref PubMed Scopus (34) Google Scholar), and was used for both high throughput screening and crystallographic studies. In the MMP IC50 assays the enzyme was used at a level at least 10-fold lower than the IC50. The initial rate of substrate hydrolysis was determined by monitoring the increase in absorbance at 412 nm on a microplate reader thermostatted at 25 °C. The % of control activity was plotted against inhibitor concentration and fit to the equation % of control activity is the inhibitor IC50 is the inhibitor concentration the rate is reduced to the and is the of the curve at of of full-length human MMP13 was expressed as the in a expression to and by it with 1 mm at for 1 h. the of inhibition by was at 25 in reaction mixtures containing 50 mm buffer (pH 7.0), 10 mm CaCl2, mm 1 mm 5,5′-dithiobis(2-nitrobenzoic acid), thioester substrate, 0.005% Brij 35, and of fit by least R.J. Google Scholar) to the equation for noncompetitive inhibition and of and Scholar), and are the of inhibitor and substrate, is the initial is the is the and is the inhibition a more we also fit the to the equation for inhibition and of and Scholar), is the by which the is the substrate is to the II mixtures to for inhibition of hydrolysis of type II collagen 50 mm buffer (pH 10 mm CaCl2, mm collagen 0.005% Brij 35, and inhibitor in Me2SO (1% final). was and the at for h. by an of by and with The protein and a with The IC50 was from a of the % of control against inhibitor protein for was expressed in E. and in a thermostatted to and with of The of of of of of of and of and was to The was with an at and was at by the of was reduced a the was to and protein expression was by to the by The in of 50 mm buffer (pH 10 mm and of The a with of at an of and a rate of was by it in of the and the 50 mm buffer (pH 10 mm 50 mm buffer (pH 10 mm 50 mm buffer (pH 10 mm The protein was dissolved by it in 50 mm buffer (pH and the at the protein in the was to and was at by 50 mm buffer (pH containing 10 mm CaCl2, 0.1 mm and an The protein was a to and in at °C. inhibitors in Me2SO in a with final). acid was to 0.1 and the 1 at °C. the protein was to and of of The for 1 and in 0.1 buffer (pH 7.0), that for in 0.1 buffer (pH several in and in a of at the on the at the in with and reduced W. 1997; PubMed Scopus Google Scholar). and initial by with J. A. Scopus Google Scholar) the of human as a search model J. G. H. Bourne Res. PubMed Scopus Google Scholar), modified for several of model Biol. 1997; PubMed Scopus Google Scholar) and the and cartilage for with and M in the of or to 10 The was and the was and for of S. Anal. Biochem. PubMed Scopus Google Scholar). the of the the cartilage was with and the in this was In by the of orally dosed to the joint tissue and inhibit degradation of cartilage, dosed orally with or of the joints with of The the joints and the was with of The type II collagen in the was by an C. H. A.J. Matrix Biol. 1999; PubMed Scopus Google Scholar). of the of an MMP13 inhibitor to prevent cartilage damage in vivo in a disease was tested in a model of OA in The and and the for this The in in an for and of to a of a of at a rate of and a of h. The a of and with a of The of joint was and of the was The joints and the to 1 the dosed orally with of or the and the joints and with to the and of cartilage as joint and of the from the the joint was in a with and on was and to the articular The was in a of normal with a to The and of the cartilage. articular of the joint was for the and for the of the a with a and a as 24 and in by was of on the at the of the of an MMP13 inhibitor to induce joint side was tested in a rat model of MSS that has been used previously to joint of the non-selective MMP (12Renkiewicz R. Qiu L. Lesch C. Sun X. Devalaraja R. Cody T. Kaldjian E. Welgus H. Baragi V. Arthritis Rheum. 2003; 48: 1742-1749Crossref PubMed Scopus (167) Google Scholar). We orally dosed with or of (i.e. the for a of A of a of of a non-selective acid MMP J.A. L.L. J. Chem. PubMed Scopus Google Scholar), as the The and and joints for of of to of MMP13 inhibitor or non-selective MMP inhibitor of joints at in and with and for to in joint the MMP13 inhibitor of inhibition the initial rate fit to the for inhibition R.J. Google Scholar). This the rate at the inhibitor and a reduced the of fit of the to the the of model was of was to the and test was used to the of the from control and at the level of The OA of at least and the and used for that the the and of MMP13 our for inhibitors of the activity of human and found 1 acid to be a inhibitor with an IC50 nm we tested 1 at to against other we found that this inhibitor not inhibit these proteases in 1 is the and selective inhibition of MMP13 by an The high selectivity of 1 and for MMP13 is in to the lack of selectivity of a MMP inhibitor known as or In addition, tested against human and in in assays cleavage of substrate was with the IC50 25 with both of these A. and M. the MMP selectivity of MMP13 selective inhibitors with that of the non-selective MMP of MMP activity was at least in inhibitor the substrate (13Weingarten H. Feder J. Anal. Biochem. 1985; 147: 437-440Crossref PubMed Scopus (90) Google was in these assays at in a was in these assays at In of the MMP selectivity which are in at as in In in buffer at and of than in buffer it was to 20 on the other of to the concentration tested in the MMP selectivity Thus, the of is to the buffer and In 1 is a noncompetitive inhibitor of and with 1 1 and a reduced of In noncompetitive inhibition the inhibitor and the substrate bind and to the enzyme at and the inhibitor with to the enzyme and to the The of substrate does not the of the inhibitor and to fit the to the rate for and inhibition in reduced of and In addition, we determined that is also a noncompetitive inhibitor of MMP13. R. it is that the or of the and is by the catalytic zinc all MMP inhibitors described to a or other that to this catalytic zinc and with substrate Opin. 2003; Google Scholar). In the noncompetitive of inhibition of MMP13 by 1 and that these inhibitors do not bind to the catalytic zinc or to the active site We that inhibition of MMP13 by is and reversible activity Furthermore, tested in an a more substrate such as type II collagen, 1 to inhibit with an IC50 nm of to noncompetitive of inhibition of MMP13 by these that they not bind to the catalytic zinc Thus, we not for MMPs to zinc inhibitors to construct to these MMP13 inhibitors determined the of of these unique MMP13 inhibitors by of MMP13 in with 1 and have been in the as and and are in for of 1 and with a of a to prevent The and for in previously described for MMP13 with zinc inhibitors S. P. H. D. J. Biol. PubMed Scopus Google Scholar, R. A. M. Tschesche H. R. W. J. Biol. 2001; PubMed Scopus Google Scholar), in the of the protein and in the A. Biochem. Biophys. Res. Commun. PubMed Scopus Google In MMP13 we have described the of the protein on the inhibitor the the G. However, it is the unique of the that is most to our drug We that the MMP13 inhibitors described here an to the that is and and crystallographic is the of unique and is the of unique and are the and the is with of of from is the of unique and is the of unique and are the and the is with of in a The acid that the is in a to the catalytic zinc 1 does not with this zinc but the of the protein and this The ester of 1 the substrate but with the that would be acid side in such as type II collagen or the synthetic or in non-selective MMP inhibitors such as This for 1 is with noncompetitive of inhibition and with the substrate inhibition for MMP inhibitors that bind to the catalytic zinc Opin. 2003; Google Scholar, S. P. H. D. J. Biol. PubMed Scopus Google Scholar, R. A. M. Tschesche H. R. W. J. Biol. 2001; PubMed Scopus Google Scholar, B. S. C. C. J.A. R. Van Wart H. Biol. 1999; PubMed Scopus Google Scholar, X. J. N. V. R. Parker D. J. Biol. PubMed Scopus Google Scholar, W. 2003; PubMed Scopus Google Scholar, H. D. A. R. W. J. Biol. Chem. 2001; Full Text Full Text PDF PubMed Scopus Google Scholar, Sci. 1999; PubMed Scopus Google Scholar). In to not the catalytic zinc ion, 1 does not the substrate of MMP13. and be by of the inhibitor and the of MMP13 in which it in 1 to the of and the In addition, the of 1 and acid and The of the ester of 1 in a with to found in of and to non-selective MMP inhibitors H. D. A. R. W. J. Biol. Chem. 2001; Full Text Full Text PDF PubMed Scopus Google Scholar, Sci. 1999; PubMed Scopus Google Scholar). The other of 1 with MMP13 are of the of the of 1 and in the MMP13 that these compounds may with the The high selectivity of 1 and for MMP13 be by of the that the of the MMPs and In most MMP for MMP13 and the is not highly in the of zinc inhibitors B. S. C. C. J.A. R. Van Wart H. Biol. 1999; PubMed Scopus Google Scholar, X. J. N. V. R. Parker D. J. Biol. PubMed Scopus Google Scholar, Sci. 1999; PubMed Scopus Google Scholar). However, we that this is in of MMP13 with 1 and and in the selectivity of inhibitor The of MMPs that the in and may be to that in MMP13 to these inhibitors. A the of these and and the other the in and are than in which may increase and may in the lack of inhibition of these the most critical is the of the in MMP13. the by in MMP13 are in a of the that is for the in and the of these these MMP13 inhibitors they have and the as MMP13. In to and to MMPs and lack at or In the side of and critical or with the of 1 and the in MMPs that may prevent these inhibitors from of the in but and and at this of these MMPs show that these may block to the W. 2003; PubMed Scopus Google Scholar). a of the MMP13 protein we have such as which are more than 1 and high selectivity for MMP13 The to the and the the and the that the active site the of highly with and for a to test inhibition of MMP13 prevent cartilage of in and in cartilage with and the of from cartilage with an IC50 that both and MMP13 are in this type of Arthritis Rheum. 2003; 48: PubMed Scopus Google Scholar) and MMPs other than may to the cleavage of this level of activity for the MMP13 inhibitor is We the of orally dosed to inhibit the degradation of type II collagen in an model of cartilage damage by the of active human MMP13 rat as as 0.1 the of the type II collagen which upon collagen degradation and cartilage Thus, by we to the joint an of an MMP13 inhibitor to the cartilage damage by MMP13. the inhibition of cartilage damage in these we to an MMP13 inhibitor prevent cartilage damage in a in vivo model of such have been In a rat model and a non-selective MMP inhibitor cartilage degradation and by Brown Taiwo Y.O. L. S.A. Full Text PDF PubMed Scopus Google Scholar). In a both and are known to be G. J. T. P. E. M. D. Google Scholar). In a model in which was by of of the a non-selective MMP inhibitor reduced cartilage damage V. J. J. A. M. Sung A. Killar L.M. T. G. R. J. Zhao W. J. J. Chem. 2003; PubMed Scopus Google Scholar). the we found that dosed orally at a level of cartilage that be in the of the cartilage from and reduced the cartilage on the and on the by and to In this a of and a (i.e. not protein of to an that is than the in of with human The efficacy by of this MMP13 inhibitor that most of the cartilage damage in this model is in due to MMP13 and the and the of a in a model be of in patients with OA. in to the level of cartilage that was in this type of model by a MMP inhibitor V. J. J. A. M. Sung A. Killar L.M. T. G. R. J. Zhao W. J. J. Chem. 2003; PubMed Scopus Google Scholar), our with show that inhibition of MMP13 is as in the cartilage area as MMP of efficacy has been for clinically MMP but safety has also been a due to the of MSS joint pathology in dosed with non-selective MMP inhibitors. the of an MMP13 inhibitor to induce MSS-like we a to described for of MMP joint (12Renkiewicz R. Qiu L. Lesch C. Sun X. Devalaraja R. Cody T. Kaldjian E. Welgus H. Baragi V. Arthritis Rheum. 2003; 48: 1742-1749Crossref PubMed Scopus (167) Google Scholar). in the of joint from MSS-like fibroplasias in of the dosed orally for with or MMP13 inhibitor In all with the non-selective MMP joint fibroplasias by of the of the and with a of in a matrix The on for the dosed with was of which to an inhibitor level that is than the in of with MMP13. an of of the acid active of Thus, the MMP13 inhibitor not induce joint side effects in this at the level to effectively articular cartilage in the model and the by the non-selective MMP By screening we have discovered an selective inhibitor of MMP13 that is unique in that it does not an we this initial and a by and we found that these compounds differ from previously MMP inhibitors in many other The noncompetitive of inhibition by 1 and is with crystallographic that that these compounds the substrate bind to the catalytic zinc Furthermore, the MMP13 selectivity of these inhibitors is is at least of more selective for inhibiting than MMP the and and the and which are also to a role in that a inhibitor of MMP13 10 that not a zinc and not inhibit or been but a acid was this to the lower selectivity and also D. R. A. R. J. Chem. Scopus Google Scholar). In MMP13 inhibitor does not bind to the catalytic zinc of it has high and and it effectively cartilage without causing joint described MMP13 inhibitors that are in to 1 or but not compounds activity B. S. R. J. O. V. Chem. Biol. 12: Full Text Full Text PDF PubMed Scopus Google Scholar). The MMP13 inhibitors 1 and are non-selective MMP inhibitors that bind to the catalytic zinc and the substrate and the of the 1 and bind in the they with in the MMP13 to selectivity. inhibitors induce to is a that is in Based on the we for the catalytic zinc in the these MMP13 inhibitors do not inhibit by causing zinc to from the The noncompetitive of inhibition that 1 and do not prevent substrate from or it is that these noncompetitive inhibitors block by to the and of this may be for the catalytic of the that the catalytic zinc as this from to and to the catalytic may be for By in and this these critical the MMP13 inhibitors may with structural or of the catalytic cause of critical active site which a in the catalytic MMP inhibitors that have been clinically have been inhibiting all or all of the known MMPs and inhibiting more such as the and the trials of non-selective MMP inhibitors have been plagued by the of musculoskeletal syndrome joint side effects in (10Clark I.M. Parker A.E. Expert Opin. Ther. Targets. 2003; 7: 19-34Crossref PubMed Scopus (75) Google Scholar). This side effect may have from to such MMP inhibitors high to Thus, MMP inhibitor has shown safety and efficacy in the The MMP13 inhibitor to have overcome these major that have the clinical development of MMP inhibitors. effectively cartilage degradation in and it does not induce joint in an model of these in we are that a major of clinical to show that such MMP13 inhibitors are of musculoskeletal syndrome side may be Based on the these compounds have in of we are that MMP13-selective inhibitors will effectively cartilage in patients without causing the MSS side effects previously with non-selective MMP inhibitors. as MMP13 activity to other such as and T. A.J. PubMed Scopus Google Scholar, H. Res. Scopus Google Scholar, R. M Ann. Rheum. Dis. PubMed Scopus Google Scholar, H. E. J. PubMed Scopus Google Scholar, D. Seyfert C. Gromnica-Ihle E. P. PubMed Scopus Google Scholar), MMP13 selective inhibitors will be in further the role of this protease in other We and for and and for and critical of this

A series of trans-tetrahydro-4-hydroxy-6-[2-(2,3,4,5-substituted-1H-pyrrol-1-yl) ethyl]-2H-pyran-2-ones and their dihydroxy acids were prepared and tested for their ability to inhibit the enzyme HMG-CoA reductase in vitro. Inhibitory potency was found to increase substantially when substituents were introduced into positions three and four of the pyrrole ring. A systematic exploration of structure-activity relationships at these two positions led to the identification of a compound ((+)-33,(+)-(4R)-trans-2-(4-fluororphenyl)-5-(1-methylethyl)-N,3- diphenyl-1- [(tetrahydro-4-hydroxy-6-oxo-2H-pyran-2-yl)ethyl]-1H-pyrrole-4- carboxamide) with five times the inhibitory potency of the fungal metabolite compactin.