MedKoo Biosciences, Inc.
Tel:   +1-919-636-5577    Fax:   +1-919-980-4831    Email:   sales@medkoo.com
Search
Login Register
Search
MedKoo Cat#: 407279 | Name: GSK484 HCl
Featured

Description:

WARNING: This product is for research use only, not for human or veterinary use.

GSK484 is a potent PAD-4 inhibitor (Protein-arginine deiminase type-4). GSK484 potently binds to the low-calcium form of PAD4 in a reversible manner (IC50 of 50 nM) and appears to be competitive with substrate. GSK484’s selectivity for PAD4 over PAD1-3 was shown in cells and also confirmed with recombinant enzymes. This probe is an inhibitor of cellular citrullination in primary neutrophils, and further phenotypic profiling has confirmed its ability to inhibit NET formation in both mouse and human neutrophils. GSK484 exhibits favourable pharmacokinetic profiles, with low-moderate clearance, and good volume of distribution and half-life in mouse and rat, and has suitable a PK profile for use as a potential in vivo tool. (http://www.thesgc.org/chemical-probes/GSK484).

Chemical Structure

GSK484 HCl
GSK484 HCl
CAS#1652591-81-5 (HCl)

Theoretical Analysis

MedKoo Cat#: 407279

Name: GSK484 HCl

CAS#: 1652591-81-5 (HCl)

Chemical Formula: C27H32ClN5O3

Exact Mass: 473.2427

Molecular Weight: 510.04

Elemental Analysis: C, 63.58; H, 6.32; Cl, 6.95; N, 13.73; O, 9.41

Price and Availability

Size Price Availability Quantity
5mg USD 150.00 Ready to ship
10mg USD 250.00 Ready to ship
25mg USD 550.00 Ready to ship
50mg USD 850.00 Ready to ship
100mg USD 1,450.00 Ready to ship
200mg USD 2,250.00 Ready to ship
500mg USD 3,250.00 Ready to ship
1g USD 4,250.00 Ready to ship
2g USD 7,450.00 2 Weeks
Show More
Bulk Inquiry
Buy Now
Add to Cart
Related CAS #
1652629-23-6 (free base) 1652591-81-5 (HCl)
Synonym
GSK484; GSK-484; GSK 484; GSK484 HCl; GSK484 hydrochloride
IUPAC/Chemical Name
((3S,4R)-3-amino-4-hydroxypiperidin-1-yl)(2-(1-(cyclopropylmethyl)-1H-indol-2-yl)-7-methoxy-1-methyl-1H-benzo[d]imidazol-5-yl)methanone hydrochloride
InChi Key
MULKOGJHUZTANI-ADMBKAPUSA-N
InChi Code
InChI=1S/C27H31N5O3.ClH/c1-30-25-20(11-18(13-24(25)35-2)27(34)31-10-9-23(33)19(28)15-31)29-26(30)22-12-17-5-3-4-6-21(17)32(22)14-16-7-8-16;/h3-6,11-13,16,19,23,33H,7-10,14-15,28H2,1-2H3;1H/t19-,23+;/m0./s1
SMILES Code
O=C(N1C[C@H](N)[C@H](O)CC1)C2=CC(OC)=C3N(C)C(C(N4CC5CC5)=CC6=C4C=CC=C6)=NC3=C2.[H]Cl
Appearance
Solid powder
Purity
>98% (or refer to the Certificate of Analysis)
Shipping Condition
Shipped under ambient temperature as non-hazardous chemical. This product is stable enough for a few weeks during ordinary shipping and time spent in Customs.
Storage Condition
Dry, dark and at 0 - 4 C for short term (days to weeks) or -20 C for long term (months to years).
Solubility
Soluble in DMSO, not in water
Shelf Life
>2 years if stored properly
Drug Formulation
This drug may be formulated in DMSO
Stock Solution Storage
0 - 4 C for short term (days to weeks), or -20 C for long term (months).
HS Tariff Code
2934.99.9001
More Info
PAD4 is a calcium-dependent enzyme which catalyses the transformation of protein arginine residues into citrulline, with the release of ammonia. PAD4-dependent citrullination/deimination of histones plays a key role in the histone code and is predicted to manifest with wide-ranging transcriptional and structural functions, including recently-discovered roles in the regulation of stem cell maintenance (1). In addition to a growing rationale in oncology, PAD4 has strong associations with multiple immune and inflammatory processes. For example, in rheumatoid arthritis the enzyme citrullinates joint proteins to break tolerance and provoke autoimmunity, with antibodies against these citrullinated epitopes (and against PAD4 itself) representing a diagnostic hallmark of the disease. In addition, PAD4 is known to promote profound chromatin decondensation during the innate immune response to infection in neutrophils by mediating formation of neutrophil extracellular traps (NETs). This is an enigmatic and exciting field where initially proposed roles for NETs in trapping pathogens for host-defence purposes (2) have been extended to demonstrate that unrestrained NETosis may be crucial for pathological deep venous thrombosis (3) ischemia/reperfusion injury (4), systemic lupus erythematosis (5), small vessel vasculitis (6) and also in rheumatoid arthritis (7). (http://www.thesgc.org/chemical-probes/GSK484).
Biological target:
GSK484 hydrochloride is a selective and reversible peptidylarginine deiminase 4 (PAD4) inhibitor that demonstrates high affinity binding to PAD4 with IC50s of 50 nM in the absence of Calcium.
In vitro activity:
Next, different concentrations of PAD4 inhibitor (GSK484) was used to treat C666-1 and 6-10B cells. The data delineated that GSK484 concentration-dependently decreased the expression of PAD4 (Fig. 3a, b). As shown in Fig. 3b, the survival fraction of NPC cells exposed to irradiation was significantly inhibited after treatment of GSK484 (Fig. 3c). The results from EdU assay showed that treatment of GSK484 suppressed cell proliferation after irradiation (Fig. 3d). Furthermore, analysis of migration and invasion revealed that treatment of GSK484 restrained the migratory and invasive ability of C666-1 cells (Fig. 3e,f). Western blot demonstrated that the citH3 protein levels were markedly reduced by PAD4 inhibition (Fig. 3g). The above findings suggested that GSK484 exerts inhibitory effects on radioresistance and aggressive phenotypes of NPC cells through inhibiting the activity of PAD4. Colony formation assay demonstrated that GSK484 treatment reversed the promotive effect of PAD4 overexpression on the survival fraction of C666-1 cells (Fig. 4c). Moreover, the effects on the increase of aggressive phenotypes of C666-1 cells caused by PAD4 were abrogated after treatment of GSK484 (Fig. 4d, f). Furthermore, in Fig. 4g, citH3 was significantly elevated in PAD4 overexpressed cells but knocked down by GSK484, suggesting that histone citrullination occurs in a PAD4-dependent way. These results suggested the key role of GSK484 in inhibiting the effects of PAD4 on radiosensitivity and malignant phenotype of NPC cells. Reference: Cell Mol Biol Lett. 2021 Mar 16;26(1):9. https://pubmed.ncbi.nlm.nih.gov/33726680/
In vivo activity:
This experiment explored whether NET release is involved in adhesion formation. GSK484 is a selective and reversible inhibitor of peptidyl arginine deiminase (PAD)-4. GSK484 was administered on day1 and 1 h before cecum cauterization. Mice treated with GSK484 presented with a significant but reduced adhesion score compared to the control mice at day 7 post-operation (Fig. 3A). Given this finding the experiment went on to evaluate if the neutrophils are responsible for these NETs, as similar phenotypes can also be produced by several other cell types, including macrophages. GSK484-treated mice displayed much lower intensities of CitH3 than control mice (Fig. 3B) and there were significantly more neutrophils undergoing NETs (stained in yellow) in the control mice than in the GSK484-treatment group (Fig. 3B). Consistent with this, ImageJ Fiji analysis revealed high correlation coefficient of colocalization between Ly-6G and CitH3 in control mice (Supplementary Fig. 2). In contrast, the correlation coefficient was low in the mice receiving GSK484 (Supplementary Fig. 2). These results suggest that GSK484 treatment protects against adhesions, likely through inhibiting NET release by neutrophils. Reference: Cell Physiol Biochem. 2021 Jul 3;55(4):400-412. https://pubmed.ncbi.nlm.nih.gov/34214389/
Solvent mg/mL mM
Solubility
DMSO 25.0 49.01
DMF 30.0 58.82
Ethanol 25.0 49.01
PBS buffer (pH 7.2) 5.0 9.80
Note: There can be variations in solubility for the same chemical from different vendors or different batches from the same vendor. The following factors can affect the solubility of the same chemical: solvent used for crystallization, residual solvent content, polymorphism, salt versus free form, degree of hydration, solvent temperature. Please use the solubility data as a reference only. Warming and sonication will facilitate dissolving. Still have questions? Please contact our Technical Support scientists.

Preparing Stock Solutions

The following data is based on the product molecular weight 510.04 Batch specific molecular weights may vary from batch to batch due to the degree of hydration, which will affect the solvent volumes required to prepare stock solutions.

Recalculate based on batch purity %
Concentration / Solvent Volume / Mass 1 mg 5 mg 10 mg
1 mM 1.15 mL 5.76 mL 11.51 mL
5 mM 0.23 mL 1.15 mL 2.3 mL
10 mM 0.12 mL 0.58 mL 1.15 mL
50 mM 0.02 mL 0.12 mL 0.23 mL
Formulation protocol:
1. Chen H, Luo M, Wang X, Liang T, Huang C, Huang C, Wei L. Inhibition of PAD4 enhances radiosensitivity and inhibits aggressive phenotypes of nasopharyngeal carcinoma cells. Cell Mol Biol Lett. 2021 Mar 16;26(1):9. doi: 10.1186/s11658-021-002512. PMID: 33726680; PMCID: PMC7962337. 2. Zeng J, Xu H, Fan PZ, Xie J, He J, Yu J, Gu X, Zhang CJ. Kaempferol blocks neutrophil extracellular traps formation and reduces tumour metastasis by inhibiting ROS-PAD4 pathway. J Cell Mol Med. 2020 Jul;24(13):7590-7599. doi: 10.1111/jcmm.15394. Epub 2020 May 19. PMID: 32427405; PMCID: PMC7339206. 3. Du M, Yang L, Gu J, Wu J, Ma Y, Wang T. Inhibition of Peptidyl Arginine Deiminase-4 Prevents Renal Ischemia-ReperfusionInduced Remote Lung Injury. Mediators Inflamm. 2020 Dec 29;2020:1724206. doi: 10.1155/2020/1724206. PMID: 33456369; PMCID: PMC7787741 4. Sudo M, Xu J, Mitani K, Jimbo M, Tsutsui H, Hatano E, Fujimoto J. Antithrombin Together with NETs Inhibitor Protected Against Postoperative Adhesion Formation in Mice. Cell Physiol Biochem. 2021 Jul 3;55(4):400-412. doi: 10.33594/000000392. PMID: 34214389.
In vitro protocol:
1. Chen H, Luo M, Wang X, Liang T, Huang C, Huang C, Wei L. Inhibition of PAD4 enhances radiosensitivity and inhibits aggressive phenotypes of nasopharyngeal carcinoma cells. Cell Mol Biol Lett. 2021 Mar 16;26(1):9. doi: 10.1186/s11658-021-002512. PMID: 33726680; PMCID: PMC7962337. 2. Zeng J, Xu H, Fan PZ, Xie J, He J, Yu J, Gu X, Zhang CJ. Kaempferol blocks neutrophil extracellular traps formation and reduces tumour metastasis by inhibiting ROS-PAD4 pathway. J Cell Mol Med. 2020 Jul;24(13):7590-7599. doi: 10.1111/jcmm.15394. Epub 2020 May 19. PMID: 32427405; PMCID: PMC7339206.
In vivo protocol:
1. Du M, Yang L, Gu J, Wu J, Ma Y, Wang T. Inhibition of Peptidyl Arginine Deiminase-4 Prevents Renal Ischemia-ReperfusionInduced Remote Lung Injury. Mediators Inflamm. 2020 Dec 29;2020:1724206. doi: 10.1155/2020/1724206. PMID: 33456369; PMCID: PMC7787741. 2. Sudo M, Xu J, Mitani K, Jimbo M, Tsutsui H, Hatano E, Fujimoto J. Antithrombin Together with NETs Inhibitor Protected Against Postoperative Adhesion Formation in Mice. Cell Physiol Biochem. 2021 Jul 3;55(4):400-412. doi: 10.33594/000000392. PMID: 34214389