2. Apoptosis NF-κB
  3. TNF Receptor RANKL/RANK Apoptosis
  4. WP9QY

WP9QY is an inhibitor targeting TNFα and RANKL, which blocks the TNFα-TNFR1 interaction and inhibits TNFα-mediated apoptosis, cytotoxicity and bone destruction. WP9QY inhibits osteoclastogenesis and promotes osteoblast differentiation, induces chondrocyte proliferation and glycosaminoglycan production, and synergizes with TGF-β3 to promote chondrogenesis. WP9QY effectively repairs full-thickness articular cartilage defects in rabbits via intra-articular injection, and inhibits methylmercury-induced reduction of NeuN-positive cells in mouse brain slices. WP9QY can be applied to the research of diseases related to methylmercury-induced neuronal death, cartilage injury, osteoarthritis and bone loss.

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WP9QY

WP9QY Chemical Structure

CAS No. : 199999-60-5

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Based on 1 publication(s) in Google Scholar

Other Forms of WP9QY:

WP9QY Related Antibodies

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TNFRSF1A/CD120a TNFRSF3/CD18 TNFRSF4 TNFRSF5/CD40 TNFRSF6/Fas/CD95 TNFRSF7/CD27 TNFRSF8/CD30 TNFRSF9/4-1BB/CD137 TNFRSF10B/DR5/CD262 TNFRSF12A/TWEAK TNFRSF16/NGF Receptor/CD271 TNFRSF18/GITR/CD357

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Description

WP9QY is an inhibitor targeting TNFα and RANKL, which blocks the TNFα-TNFR1 interaction and inhibits TNFα-mediated apoptosis, cytotoxicity and bone destruction. WP9QY inhibits osteoclastogenesis and promotes osteoblast differentiation, induces chondrocyte proliferation and glycosaminoglycan production, and synergizes with TGF-β3 to promote chondrogenesis. WP9QY effectively repairs full-thickness articular cartilage defects in rabbits via intra-articular injection, and inhibits methylmercury-induced reduction of NeuN-positive cells in mouse brain slices. WP9QY can be applied to the research of diseases related to methylmercury-induced neuronal death, cartilage injury, osteoarthritis and bone loss[1][2][3][4].

In Vitro

WP9QY-functionalized PEG hydrogels (100-250 μM; 24 h TNFα exposure, 6 d NGF differentiation culture) protect differentiated PC12 cells from apoptosis induced by 50 ng/mL human TNFα, maintain cell viability, and prevent the elevation of caspase 3/7 activation levels[1].
WP9QY-functionalized PEG hydrogel (100 μM; 24 h) protects encapsulated mouse islets from 50 ng/mL human TNFα-induced apoptosis and impaired insulin secretion, and maintains cell viability and function[1].
WP9QY (10 μM; 72 h) inhibits neuronal cell death induced by 50 μM methylmercury in cultured mouse organotypic brain slices in vitro[2].
WP9QY (100 μM; 14 d) significantly enhances chondrogenic differentiation of hMSCs in a two-dimensional culture system via accumulation of glycosaminoglycan matrix, as confirmed by Alcian Blue staining (HY-D0001) and absorbance measurement[3].
WP9QY (25-100 μM; 72 h) dose-dependently promotes the proliferation of hMSCs cultured in maintenance medium, chondrocyte differentiation medium, and osteoblast differentiation medium, and this result is confirmed by Hoechst 33342 (HY-15559) fluorescence assay[3].
WP9QY (100 μM; 14 d) induces chondrocyte differentiation in both control and RANKL-knockdown C3H10T1/2 cells in a RANKL signaling-independent manner[3].
WP9QY (5-50 μM; 20 min) dose-dependently inhibits RANKL-induced signaling pathways in RAW 264.7 cells, without affecting M-CSF-induced signaling pathways[4].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

WP9QY Related Antibodies

Cell Viability Assay[1]

Cell Line: differentiated rat adrenal pheochromocytoma (PC12) cells
Concentration: 100-250 μM WP9QY; 50 ng/mL human TNFα; 50 ng/mL 2.5S NGF
Incubation Time: 24 hours (TNFα exposure); 6 days (NGF differentiation)
Result: Maintained viability comparable to unchallenged cells in PEG-RGD-WP9QY hydrogels (100 μM and 250 μM WP9QY) under TNFα challenge.
Showed no significant increase in caspase 3/7 activity.
Confirmed minimal cell death via Live/Dead imaging.

Cell Differentiation Assay[3]

Cell Line: human mesenchymal stem cells (hMSCs)
Concentration: 100 μM
Incubation Time: 14 days
Result: Increased absorbance at 620 nm to ~0.27, which was significantly higher than control's ~0.17 with p < 0.01.

Cell Differentiation Assay[3]

Cell Line: mouse chondrogenic ATDC5 cells
Concentration: 25-100 μM
Incubation Time: 18 days
Result: Induced dose-dependent chondrocytic differentiation, with more intense Alcian Blue staining at 100 μM compared to 25 μM, and no staining in untreated control cells.

Cell Differentiation Assay[3]

Cell Line: human mesenchymal stem cells (hMSCs)
Concentration: 50-150 μM (with 10 ng/mL TGF-β3 present throughout culture)
Incubation Time: 14 days
Result: Induced dose-dependent enlargement of hMSC 3D pellets, with larger pellets observed at 150 μM compared to 50 μM.
Increased Alcian Blue-positive cartilage matrix production in treated pellets.

Cell Differentiation Assay[3]

Cell Line: human mesenchymal stem cells (hMSCs)
Concentration: 100 μM (tested alone and in combination with 20 ng/mL TGF-β3)
Incubation Time: 15 days
Result: Strongly enhanced chondrogenesis alone, while TGF-β3 alone weakly enhanced chondrogenesis.
Combined treatment resulted in synergistically increased Alcian Blue staining intensity, showing greater chondrogenesis than either treatment alone.

Cell Differentiation Assay[3]

Cell Line: human mesenchymal stem cells (hMSCs)
Concentration: 100 μM (tested alone and in combination with 10 ng/mL TGF-β3)
Incubation Time: 21 days
Result: Increased pellet size and induced cartilage matrix production (positive Alcian Blue and Safranin O staining) alone.
TGF-β3 alone increased chondrocytic differentiation but did not affect pellet size.
Combined treatment resulted in synergistically enhanced cartilage matrix production and larger pellet size compared to either treatment alone.

Cell Proliferation Assay[3]

Cell Line: human mesenchymal stem cells (hMSCs)
Concentration: 25-100 μM
Incubation Time: 72 hours
Result: Promoted hMSC proliferation in maintenance, chondrocyte-differentiation, and osteoblast-differentiation media in a dose-dependent manner.
Elevated fluorescence intensity at 25 μM compared to untreated controls; further increased intensity at 100 μM, with p < 0.01 for all comparisons to untreated controls.

Cell Differentiation Assay[3]

Cell Line: mouse mesenchymal C3H10T1/2 cells (control and RANKL-knockdown)
Concentration: 100 μM
Incubation Time: 14 days
Result: Enhanced chondrocytic differentiation (positive Alcian Blue staining) in both control and RANKL-knockdown C3H10T1/2 cells.
RANKL knockdown had no effect on WP9QY-induced chondrogenesis.
In Vivo

WP9QY (5 mg; intraarticular; 2 doses (days 14 and 21)) significantly improves repair of full-thickness articular cartilage defects in rabbits, as indicated by a reduced modified Wakitani score[3].
WP9QY (2.1 mg/kg/d; continuous infusion via osmotic minipump; 4 weeks) completely prevents ovariectomy-induced osteoporosis in female C57BL/6J mice, restoring cancellous bone parameters and reducing osteoclast activity to near sham-operated levels[4].
WP9QY (9 mg/kg; s.c.; every 3 hours; 48 hours) completely prevents low dietary calcium2+-induced osteoporosis in male CD-1 mice, restoring cancellous bone parameters and reducing osteoclast activity to near normal-calcium diet levels[4].
WP9QY (4 mg/kg/d; continuous infusion via osmotic minipump; 7 days) completely prevents low dietary calcium2+-induced osteoporosis in both TNFR(I)-/- TNFR(II)-/- and wild-type mice[4].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Animal Model: Japanese white (male, 19-20 weeks old)[3]
Dosage: 5 mg
Administration: intraarticular; 2 doses (days 14 and 21)
Result: Caused partial filling of articular cartilage defects and expanded Safranin-O-positive regions in defects.
Reduced mean modified Wakitani score to ~7, versus ~12 for vehicle (P=0.04).
Animal Model: C57BL/6J Mice (female, 12 weeks old, bilateral ovariectomy-induced)[4]
Dosage: 2.1 mg/kg/d
Administration: continuous infusion via osmotic minipump; 4 weeks
Result: Maintained bone mineral density of proximal tibiae at levels not significantly different from sham-operated mice.
Restored cancellous bone volume fraction from 9.10% to 14.79%.
Restored trabecular number from 6.10 no./mm to 8.01 no./mm.
Restored trabecular spacing from 150.12 μm to 109.998 μm.
Reduced osteoclast number per bone surface from 7.62 no./mm to 3.65 no./mm.
Reduced osteoclast surface per bone surface from 11.58% to 5.96%.
Blocked the ovariectomy-induced increase in urinary deoxypyridinoline cross-links.
Had no effect on body weight or uterine weight.
Animal Model: TNFR(I)-/- TNFR(II)-/- Mice (129/B6 background); wild-type (genetically matched, low dietary calcium-induced)[4]
Dosage: 4 mg/kg/d
Administration: continuous infusion via osmotic minipump; 7 days
Result: Maintained bone mineral density at levels not significantly different from mice on a normal-calcium diet in both TNFR double-knockout and wild-type mice.
Blocked the low-calcium diet-induced increase in urinary deoxypyridinoline cross-links in both genotypes.
Molecular Weight

1226.38

Formula

C58H71N11O15S2
CAS No.
Appearance

Solid

Color

White to off-white

Sequence

Tyr-Cys-Trp-Ser-Gln-Tyr-Leu-Cys-Tyr (Disulfide bridge:Cys2-Cys8)
Sequence Shortening

YCWSQYLCY (Disulfide bridge:Cys2-Cys8)
Shipping

Room temperature in continental US; may vary elsewhere.
Storage

Sealed storage, away from moisture

Powder -80°C 2 years
-20°C 1 year

*In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture)

Solvent & Solubility
In Vitro: 

DMSO : 100 mg/mL (81.54 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

H2O : 1 mg/mL (0.82 mM; Need ultrasonic)

Preparing
Stock Solutions
Concentration Solvent Mass 1 mg 5 mg 10 mg
1 mM 0.8154 mL 4.0770 mL 8.1541 mL
5 mM 0.1631 mL 0.8154 mL 1.6308 mL
View the Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

  • Molarity Calculator

  • Dilution Calculator

Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

Mass
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Concentration
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Volume
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Molecular Weight *

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

This equation is commonly abbreviated as: C1V1 = C2V2

Concentration (start)

C1

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Volume (start)

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Concentration (final)

C2

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Volume (final)

V2

In Vivo:

Select the appropriate dissolution method based on your experimental animal and administration route.

For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

  • Protocol 1

    Add each solvent one by one:  2% DMSO    10% PEG300    2% Tween-80    86% Saline

    Solubility: ≥ 2 mg/mL (1.63 mM); Clear solution

In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:

Dosage

mg/kg

Animal weight
(per animal)

g

Dosing volume
(per animal)

μL

Number of animals

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
%
DMSO +
% 
+
%
Tween-80 +
%
Saline
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Calculation results:
Working solution concentration: mg/mL
Method for preparing stock solution: mg drug dissolved in μL  DMSO (Stock solution concentration: mg/mL).

*In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture)

The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only. If necessary, please contact MedChemExpress (MCE).
Method for preparing in vivo working solution for animal experiments: Take μL DMSO stock solution, add μL . μL , mix evenly, next add μL Tween 80, mix evenly, then add μL Saline.
 If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
Purity & Documentation
References

Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
DMSO 1 mM 0.8154 mL 4.0770 mL 8.1541 mL 20.3852 mL
5 mM 0.1631 mL 0.8154 mL 1.6308 mL 4.0770 mL
10 mM 0.0815 mL 0.4077 mL 0.8154 mL 2.0385 mL
15 mM 0.0544 mL 0.2718 mL 0.5436 mL 1.3590 mL
20 mM 0.0408 mL 0.2039 mL 0.4077 mL 1.0193 mL
25 mM 0.0326 mL 0.1631 mL 0.3262 mL 0.8154 mL
30 mM 0.0272 mL 0.1359 mL 0.2718 mL 0.6795 mL
40 mM 0.0204 mL 0.1019 mL 0.2039 mL 0.5096 mL
50 mM 0.0163 mL 0.0815 mL 0.1631 mL 0.4077 mL
60 mM 0.0136 mL 0.0680 mL 0.1359 mL 0.3398 mL
80 mM 0.0102 mL 0.0510 mL 0.1019 mL 0.2548 mL

* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

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Help & FAQs
  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

Keywords:

WP9QY199999-60-5TNF ReceptorRANKL/RANKApoptosisTumor Necrosis Factor ReceptorTNFRhuman mesenchymal stem cellRAW 264.7 cellsmouse pancreatic isletsTNFαC57BL/6J miceRANKLATDC5 cellsPC12 cellsC3H10T1/2 cellsprimary murine bone marrow cellsInhibitorinhibitorinhibit

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