| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| 250mg |
|
||
| 500mg | |||
| Other Sizes |
Purity: ≥98%
Diquafosol tetrasodium (Diquas; INS-365) is a novel and potent P2Y2 purinoceptor agonist that stimulates fluid and mucin secretion on the ocular surface, and is used as a topical treatment of dry eye disease.
| Targets |
P2Y2 receptor
Diquafosol Tetrasodium is an agonist of the purinergic receptor P2Y₂ (P2Y2 receptor). [1] |
|---|---|
| ln Vitro |
Cell viability was significantly reduced after treatment with 30% diquafosol tetrasodium for 1 hour and 10% and 20% diquafosol tetrasodium for 6 hours. 24 hours after monolayer cell damage, wound healing was significantly aggravated by 3% diquafosol tetrasodium and 0.3% HCEC [1].
Diquafosol Tetrasodium (at 3% concentration) showed a significant, time-dependent inhibitory effect on the proliferation of Human Corneal Epithelial Cells (HCECs) and increased cytotoxicity compared to hyaluronic acid treatments. Cell viability significantly decreased after 1 hour of treatment with 30% diluted (i.e., 0.9% final concentration?) diquafosol and after 6 hours of treatment with 10% and 20% diluted diquafosol. [1] Diquafosol Tetrasodium increased lactate dehydrogenase (LDH) activity (a marker of cell membrane damage) at all exposure times (1, 6, 24 hours) compared to hyaluronic acid, with a significant increase after 24 hours. [1] Phase-contrast microscopy showed that HCECs treated with Diquafosol Tetrasodium (3%) detached more from the culture dish compared to cells treated with hyaluronic acid. [1] Transmission electron microscopy revealed that HCECs treated with 3% Diquafosol Tetrasodium exhibited loss of microvilli, formation of large cytoplasmic vacuoles (likely due to swollen organelles), and condensation of nuclear chromatin along the nuclear periphery, indicating degenerative changes. [1] In a scratch wound healing assay, Diquafosol Tetrasodium (3%) significantly stimulated re-epithelialization of scratched HCECs after 24 hours, although the effect was less pronounced compared to hyaluronic acid treatments. [1] The commercial 3% Diquafosol Tetrasodium ophthalmic solution contained benzalkonium chloride (BAC) as a preservative at a concentration of 0.002%. [1] |
| ln Vivo |
In a mini-dry eye model, the P2Y2 agonist tetraphosphonate was found to restore surface health based on increased tear production, corneal epithelial resistance, and the release of glycoproteins from goblet cells. Beginning in week 2 and continuing for 2 weeks, the maximum decrease in dye-extrasigma rate was approximately 50% at doses as low as 1% of diquafosol tetrasodium [2]. Diquafosol tetrasodium can significantly interfere with potential damage when the concentration exceeds 0.1% w/v [3].
|
| Cell Assay |
The MTT assay is used to assess the viability of human corneal epithelial cells (HCECs). Diquafosol (100%, 20%, or 30% diluted) or DMEM (100 mL) is added to controls when the cells are subconfluent. To get rid of the medications, plates are cleaned three times using PBS after one, six, and twenty-four hours. After a 24-hour incubation period, cell viability is assessed. Next, each well receives an addition of MTT. After 4 hours of dark incubation at 37°C, the samples are removed from the media. The precipitates are reconstituted in DMSO. At 570 nm, absorbances are measured using a plate reader[1].
Cell viability was assessed using the MTT assay. HCECs were seeded in 96-well plates and allowed to grow until subconfluent. The cells were then treated with dilutions of 3% Diquafosol Tetrasodium (10%, 20%, 30% dilutions, corresponding to final concentrations of 0.3%, 0.6%, 0.9%?) or hyaluronic acid solutions for 1, 6, or 24 hours. After treatment and washing, MTT reagent was added, and the cells were incubated. The resulting formazan crystals were dissolved in DMSO, and absorbance was measured at 570 nm. [1] Cytotoxicity was assessed using the lactate dehydrogenase (LDH) leakage assay. HCECs were seeded in 96-well plates and treated similarly. Supernatants were collected, and LDH activity was measured using a cytotoxicity assay kit. Absorbance was read at 490 nm. [1] Cell morphology was observed using inverted phase-contrast light microscopy after treating HCECs with the drugs for 24 hours. [1] Transmission electron microscopy was performed to examine ultrastructural changes. HCECs grown to confluence were treated with the drugs for 4 and 8 hours, then fixed, post-fixed, dehydrated, embedded, sectioned, stained, and examined. [1] A scratch wound healing assay was performed. HCECs were grown to confluence on collagen-coated chamber slides, scratched with a pipette tip, washed, and then incubated with the drugs for 24 hours. Wound widths were measured before and after incubation, and the percentage of wound closure was calculated. [1] |
| Animal Protocol |
Rats: The Schirmer test score was at least 50% lower in an SD rat dry eye model after exorbital lacrimal gland extraction. INS365-containing eye drops are administered six times a day for the following four weeks at concentrations ranging from 0.03% to 3.0% after eight weeks, at which point there have been noticeable increases in corneal epithelial permeability. Measurements of fluorescein penetrance obtained with a modified anterior fluorometer at 1, 2, and 4 weeks following initial application are used to assess the function of the corneal barrier. The periodic acid-Schiff reagent (PAS)-stained area is assessed in histologic sections of the bulbar and tarsal conjunctiva following INS365 application[2].
|
| Toxicity/Toxicokinetics |
3% diquinofoside tetrasodium eye drops (containing 0.002% benzalkonium chloride) showed significant cytotoxicity to human corneal endothelial cells (HCECs) in vitro. This cytotoxicity was time- and concentration-dependent, leading to decreased cell viability, increased lactate dehydrogenase (LDH) leakage, cell shedding, and ultrastructural damage (disappearance of microvilli, vacuolization, and chromatin condensation). Studies have shown that the observed cytotoxicity may be primarily attributed to the preservative benzalkonium chloride (BAC) present in the formulation. [1] The electrolyte composition, pH, and osmolarity of the 3% diquinofoside tetrasodium solution were determined: Na⁺ 211.8 mEq/L, K⁺ 19.3 mEq/L, Cl⁻ 99 mEq/L, pH 7.0, and osmolarity 297 mOsm/kg. Some of these values (e.g., Na⁺, K⁺) are outside the ideal physiological range for extracellular fluid. [1]
|
| References |
|
| Additional Infomation |
Dequafosol tetrasodium is an ophthalmic solution used to treat dry eye. It acts as a P2Y₂ receptor agonist, promoting the secretion of tears and mucins, rather than simply replenishing tears. [1]
This study compared commercially available 3% dequafosol tetrasodium eye drops (containing 0.002% benzalkonium chloride) with preservative-free hyaluronic acid eye drops. The results indicated that the presence of the preservative benzalkonium chloride was likely a significant cause of the observed epithelial toxicity. The authors suggest that preservative-free dequafosol formulations are safer for long-term use. [1] |
| Molecular Formula |
C18H22N4O23P4-4.4[NA+]
|
|---|---|
| Molecular Weight |
878.234680000001
|
| Exact Mass |
877.92
|
| Elemental Analysis |
C, 24.62; H, 2.53; N, 6.38; Na, 10.47; O, 41.90; P, 14.11
|
| CAS # |
211427-08-6
|
| Related CAS # |
211427-08-6 (tetrasodium); 59985-21-6
|
| PubChem CID |
148196
|
| Appearance |
White to off-white solid powder
|
| Hydrogen Bond Donor Count |
6
|
| Hydrogen Bond Acceptor Count |
23
|
| Rotatable Bond Count |
14
|
| Heavy Atom Count |
53
|
| Complexity |
1460
|
| Defined Atom Stereocenter Count |
8
|
| SMILES |
C1=CN(C(=O)NC1=O)[C@H]2[C@@H]([C@@H]([C@H](O2)COP(=O)([O-])OP(=O)([O-])OP(=O)([O-])OP(=O)([O-])OC[C@@H]3[C@H]([C@H]([C@@H](O3)N4C=CC(=O)NC4=O)O)O)O)O.[Na+].[Na+].[Na+].[Na+]
|
| InChi Key |
OWTGMPPCCUSXIP-FNXFGIETSA-J
|
| InChi Code |
InChI=1S/C18H26N4O23P4.4Na/c23-9-1-3-21(17(29)19-9)15-13(27)11(25)7(41-15)5-39-46(31,32)43-48(35,36)45-49(37,38)44-47(33,34)40-6-8-12(26)14(28)16(42-8)22-4-2-10(24)20-18(22)30;;;;/h1-4,7-8,11-16,25-28H,5-6H2,(H,31,32)(H,33,34)(H,35,36)(H,37,38)(H,19,23,29)(H,20,24,30);;;;/q;4*+1/p-4/t7-,8-,11-,12-,13-,14-,15-,16-;;;;/m1..../s1
|
| Chemical Name |
tetrasodium;[[(2R,3S,4R,5R)-5-(2,4-dioxopyrimidin-1-yl)-3,4-dihydroxyoxolan-2-yl]methoxy-oxidophosphoryl] [[[(2R,3S,4R,5R)-5-(2,4-dioxopyrimidin-1-yl)-3,4-dihydroxyoxolan-2-yl]methoxy-oxidophosphoryl]oxy-oxidophosphoryl] phosphate
|
| Synonyms |
INS-365; DE-089; KPY-998; INS 365; DE 089; KPY 998; INS365; DE089; KPY998; Diquas
|
| HS Tariff Code |
2934.99.9001
|
| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| Solubility (In Vitro) |
H2O: ~100 mg/mL (~113.9 mM)
DMSO: ~1 mg/mL (~1.1 mM) |
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: 100 mg/mL (113.87 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.
(Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.1387 mL | 5.6933 mL | 11.3865 mL | |
| 5 mM | 0.2277 mL | 1.1387 mL | 2.2773 mL | |
| 10 mM | 0.1139 mL | 0.5693 mL | 1.1387 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
Evaluation of efficacy of diquafosol sodium ophthalmic solution and sodium hyaluronate ophthalmic solution on treatment of dry eye (multicenter study).
CTID: UMIN000009071
Phase: Status: Complete: follow-up complete
Date: 2012-11-01
|
|
|
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
COA
