Sid-09-ipc-poster-v2-4-27-final.ppt

Isoprenylcysteine (IPC) analogs: a novel class of NSAIDs show activity
to different inflammatory endpoints including reduced erythema*
E. Perez1, J. B. Stock1, 2, A. V. Gonzalez1, K. Rouzard1, S.Y. Lee1, M.V. Voronkov1, K. Rapole1, J. Barrero-Oro1,J. S. Gordon1,
M. B. Stock1 1Signum Biosciences, Monmouth Junction, NJ. 2Molecular Biology, Princeton University, Princeton, NJ.
Introduction
Fig. 3 Synthesing 2nd Generation IPC
Fig. 5 IPCs More Active than Current
Using mouse models for acute dermal inflammation and allergic contact Analogs for Pharmaceutical Development
Topical Anti-Rosacea Agents vs
hypersensitivity we have previously demonstrated that the isoprenylcysteine(IPC) analog, N-acetyl-S-farnesyl-L-cysteine (AFC) is an effective topical Neutrophil Infiltration
anti-inflammatory in vivo comparable in activity to glucocorticoids and 2nd generation analogs:
standard non-steroidal anti-inflammatory drugs (NSAIDs), but lower in potency (J.S. Gordon et al, ‘08, JID, 128: 643). To assess the anti-
inflammatory potential of novel IPC analogs, a total of 126 chemically-diverse IPC analogs were synthesized with structural modifications made to four distinct regions: 1) the cysteine
-carboxyl, 2) the farnesyl side chain, 3) the sulfur and 4) the cysteine
-amino. Analogs were tested for three endpoints in mice: inhibition of TPA-induced ear thickness (edema), neutrophil % Inhibition
infiltration (myeloperoxidase - MPO) and erythema (redness). Several Using the naturally occurring farnesyl-cysteine structure, chemical analogs substantially more potent than AFC were identified for all three modifications are made to four distinct regions of the molecule to generate endpoints. Thus, IPC analogs could yield novel pharmaceutical leads for the novel IPC compounds. Compounds are screened in in vitro cell-based treatment of specific inflammatory skin conditions. Our lead pharmaceutical assays and then in vivo for anti-inflammatory activity using the mouse ear No Activity
candidate SIG989 is currently being developed as an anti-rosacea AZA (15%)
Metro (1%)
Metro (4%)
Clonidine
SIG989 (4%) SIG1176 (4%) SIG1191 (4%)
Fig. 1 IPC Analogs Structural Mimics of G-
Fig. 4 IPCs More Active than Current
Using a mouse ear model of TPA-induced inflammation, compounds weretested for activity measuring inhibition of myeloperoxidase (in tissue a Protein C-Terminus
Topical Anti-Rosacea Agents vs Erythema
marker for neutrophil infiltration in response to irritation) compared to currenttopical anti-rosacea agents (azelaic acid (AZA), metronidazole (Metro) andclonidine). Results demonstrate IPC analogs are 2 to 3 times more active in reducing chemically-induced neutrophil infiltration. Mice received 1.2
g/20
l TPA to each ear then the same volume of compound in EtOH after 5 minutes. 6 mm ear punches were taken and MPO measurements taken 24 (e.g. Ras, Rac, Rho, Rab,
G-protein -subunits)
hours after TPA treatment. All compounds were tested at the concentration % Inhibition
Fig. 6 IPCs More Active than Current
Topical Anti-Rosacea Agents vs Edema
G-proteins participate in eliciting inflammatory responses such as the release of pro-inflammatory mediators, and the migration and activation of inflammatory cells. Located near the end of each G-protein is a conserved AZA (15%)
Metro (1%)
Metro (4%)
Clonidine
SIG989 (4%) SIG1176 (4%) SIG1191 (4%)
cysteine residue modified with a prenyl tail (either 15 or 20 carbon side- chain). IPC analogs are structural mimics of the lipidated C-termini of the G Using a mouse ear model of TPA-induced inflammation, compounds were subunit of all heterotrimeric G-proteins, as well as that of small molecular tested for activity measuring for the reduction of erythema and compared to weight GTPases such as Ras, Rho and Rac.
current topical anti-rosacea agents (azelaic acid (AZA), metronidazole (Metro) and clonidine). Results demonstrate IPC analogs are ~4-times more Fig. 2 IPC Analogs Mechanism of Action
active in reducing chemically-induced erythema. Mice received 1.2
g/20
l % Inhibition
TPA to each ear then the same volume of compound in EtOH after 5 minutes. Erythema measurements were taken at 24 hours after TPA 



treatment using a Konica Minolta CR400 and all compounds were tested in AZA (15%)
Metro (1%)
Metro (4%)
Clonidine
SIG989 (4%) SIG1176 (4%) SIG1191 (4%)


Summary/Conclusions
 


Using a mouse ear model of TPA-induced inflammation, compounds were  

Topical NSAID technology modulates G-protein signaling pathways by tested for activity measuring inhibition of edema compared to current topical anti-rosacea agents (azelaic acid (AZA), metronidazole (Metro) and 



clonidine). Results demonstrate IPC analogs are 3 to 4 times more active in 



reducing chemically-induced neutrophil infiltration. Mice received 1.2
g/20
l 


Novel, more potent IPC analogs are generated by introducing modifications to four distinct sites of the naturally occurring farnesyl- TPA to each ear then the same volume of compound in EtOH after 5 IPC analogs through their novel mechanism of action successfully modulate minutes. 6 mm ear punches were taken and weighed 24 hours after TPA inflammatory signaling by binding to GPCR prenyl-binding pockets, normally treatment. All compounds were tested at the concentration shown.
used by G-proteins to activate inflammation signaling pathways. Several other
SIG989 as well as other IPC analogs exhibit superior activity to current important signaling proteins, in addition to GPCRs, have recently been topical rosacea and anti-erythema agents (metronidazole, azelaic acid identified to possess prenyl-binding pockets or require a prenyl tail for binding *Work supported in part by NIH SBIR grant No. 1R43AI06034-01A2 representing other potential targets for IPC compounds.

Source: http://www.signumbiosciences.com/pdf/SID-IPC%20Analogues%20against%20current%20rosacea%20treatment.pdf