PROTAC ALK5 Degrader-1
PROTAC ALK5 Degrader-1 is a selective ALK5 PROTAC degrader. PROTAC ALK5 Degrader-1 induces ALK5 degradation via ALK5 ATP-binding pocket engagement, CRBN recruitment, and the ubiquitin-proteasome system.PROTAC ALK5 Degrader-1 inhibits ALK5 downstream signaling. PROTAC ALK5 Degrader-1 can be used for the research of pulmonary fibrosis.
(Pink: Anaplastic lymphoma kinase (ALK) ligand (HY-183441); Blue: Cereblon ligand (HY-14658); Black: linker (HY-183440)).
For research use only. We do not sell to patients.
- CAS No.: 3125240-35-6
- Formula: C43H43ClFN9O7
- Molecular Weight:852.31
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Storage:
Please store the product under the recommended conditions in the Certificate of Analysis.
All PROTACs Isoforms
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Biological Activity
Description
In Vitro
PROTAC ALK5 Degrader-1 (Compound A31) (0.1-1 μM; 24 h) induces ALK5 degradation in NIH3T3 cells[1].
PROTAC ALK5 Degrader-1 (1 μM; 2-48 h) induces time-dependent ALK5 degradation in NIH3T3 cells[1].
PROTAC ALK5 Degrader-1 (0.37-30 μM; 24 h) induces concentration-dependent ALK5 degradation in NIH3T3 cells over 24 h[1].
PROTAC ALK5 Degrader-1 (1 μM; 48 h) selectively induces ALK5 degradation over the related ALK4 receptor in NIH3T3 cells[1].
PROTAC ALK5 Degrader-1 (1 μM; 48 h)-induced ALK5 degradation in NIH3T3 cells is reversible, with gradual recovery of ALK5 protein levels following compound wash-out after 48 h of 1 μM treatment[1].
PROTAC ALK5 Degrader-1 (1 μM; 48 h)-induced ALK5 degradation in NIH3T3 cells requires engagement of the ALK5 ATP-binding pocket, CRBN recruitment, and the ubiquitin-proteasome system[1].
PROTAC ALK5 Degrader-1 (1 μM; 48 h) suppresses phosphorylation of ALK5 downstream effectors Smad2/3, AKT, and ERK1/2 in TGF-β1-stimulated NIH3T3 cells[1].
PROTAC ALK5 Degrader-1 (0.2-5 μM; 48 h) dose-dependently inhibits TGF-β1-induced upregulation of fibrotic markers Collagen I and α-SMA in NIH3T3 cells[1].
PROTAC ALK5 Degrader-1 (0.2-5 μM; 48 h) dose-dependently reduces the fluorescence intensity of fibrotic markers Collagen I and α-SMA in TGF-β1-stimulated NIH3T3 cells following 1 h of TGF-β1 stimulation[1].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.
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Cell Line:NIH3T3
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Concentration:0.1 μM; 1 μM
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Incubation Time:24 h
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Result:Induced 18% ALK5 degradation at 0.1 μM.
Induced 49% ALK5 degradation at 1 μM.
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Cell Line:NIH3T3
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Concentration:1 μM
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Incubation Time:2 h; 4 h; 8 h; 12 h; 24 h; 48 h
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Result:Detected reduction in ALK5 levels starting at 24 h.
Showed progressive degradation over time, reaching maximal effect at 48 h.
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Cell Line:NIH3T3
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Concentration:0.37 μM; 1.1 μM; 3.3 μM; 10 μM; 30 μM
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Incubation Time:24 h
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Result:Elicited ALK5 degradation at 0.37 μM.
Showed most pronounced degradation effect at 1.1 μM.
Exhibited reduced degradation efficiency at concentrations above 1.1 μM, consistent with a hook effect-like phenomenon.
Did not achieve 50% degradation within the tested concentration range.
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Cell Line:NIH3T3
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Concentration:1 μM
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Incubation Time:48 h
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Result:Efficiently degraded ALK5.
Barely degraded ALK4 under the same experimental conditions.
Kept ALK5 protein levels suppressed under continuous treatment.
Showed gradual recovery of ALK5 protein levels after compound wash-out.
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Cell Line:NIH3T3
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Concentration:1 μM
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Incubation Time:48 h
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Result:Significantly reduced phosphorylation of Smad2/3, AKT, and ERK1/2.
Did not alter total protein levels of Smad2/3, AKT, and ERK1/2.
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Cell Line:NIH3T3
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Concentration:0.2 μM; 1 μM; 5 μM
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Incubation Time:48 h
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Result:Dose-dependently attenuated the TGF-β1-induced increase in Collagen I protein expression.
Dose-dependently attenuated the TGF-β1-induced increase in α-SMA protein expression.
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Cell Line:NIH3T3
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Concentration:0.2 μM; 1 μM; 5 μM
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Incubation Time:48 h
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Result:Dose-dependently reduced the fluorescence intensity of Collagen I.
Dose-dependently reduced the fluorescence intensity of α-SMA.
Confirmed attenuation of TGF-β1-induced fibroblast activation.
Parmacokinetics
| Species | Dose | Route | AUC0-t | AUC0-∞ | MRT0-t | MRT0-∞ | T1/2 | Tmax | Cmax | F |
|---|---|---|---|---|---|---|---|---|---|---|
| Mice[1] | 1 mg/kg | i.v. | 264.94 μg/L·h | 263.61 μg/L·h | 0.48 h | 0.50 h | 1.33 h | 0.08 h | 678.84 μg/L | / |
| Mice[1] | 10 mg/kg | p.o. | 7.36 μg/L·h | 7.50 μg/L·h | 0.36 h | 0.48 h | 0.50 h | 0.08 h | 28.67 μg/L | 0.28 % |
| Mice[1] | 10 mg/kg | i.p. | 1869.79 μg/L·h | 1875.37 μg/L·h | 1.67 h | 1.75 h | 4.24 h | 0.33 h | 1139.89 μg/L | 70.57 % |
In Vivo
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
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Animal Model:C57BL/6 (Bleomycin-induced pulmonary fibrosis model)[1]
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Dosage:15 mg/kg; 30 mg/kg
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Administration:i.p.; daily; 14 days
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Result:Alleviated Bleomycin-induced body weight loss compared to vehicle-treated mice.
Reduced lung index by 10.6% at 15 mg/kg relative to dose-matched TP008.
Reduced fibrosis score by 9.8% at 15 mg/kg relative to dose-matched TP008, with efficacy comparable to nintedanib.
Reduced collagen positive percentage by 15.9% at 15 mg/kg relative to dose-matched TP008, restoring collagen levels toward sham control at both doses.
Reduced Collagen I fluorescence intensity by 15% and α-SMA fluorescence intensity by 20% at 15 mg/kg relative to dose-matched TP008, with efficacy comparable to nintedanib at both doses.
Reduced the proportion of lung dendritic cells (CD11c+ MHCII+) with greater efficacy than nintedanib, and attenuated bleomycin-induced accumulation of F4/80+ CD11b+ CD206+ macrophages with effects comparable to nintedanib, in a dose-dependent manner.
Reduced lung tissue ALK5 protein levels and suppressed downstream signaling activation (p-Smad2/3, p-AKT, p-ERK1/2) in a dose-dependent manner; showed greater suppression of p-AKT (additional 27%) and p-ERK1/2 (additional 79%) at 15 mg/kg relative to dose-matched TP008.
Showed no significant systemic toxicity: histopathology of heart, liver, spleen, and kidneys showed normal tissue architecture, and serum ALT/AST levels were not significantly different from sham controls.
Chemical Information
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CAS No. 3125240-35-6
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Molecular Weight 852.31
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Formula C43H43ClFN9O7
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SMILES
ClC1=CC(C2=CC(N3C4=CC=NC=C4N(CC(NCCN5CCN(CCCCOC6=CC=CC(C(N7C8C(NC(CC8)=O)=O)=O)=C6C7=O)CC5)=O)C3=O)=C(C)C=N2)=C(F)C=C1
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Shipping
Room temperature in continental US; may vary elsewhere.
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Storage
Please store the product under the recommended conditions in the Certificate of Analysis.
Protocols
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Fibrosis/Collagen Morphometry
Fibrosis and collagen morphometry is based on the quantitative visualization of fibrillar collagen deposition in tissue sections using histochemical stains such as Sirius Red (Picrosirius Red) or Masson's trichrome, followed by image-based or polarization-enhanced analysis to estimate collagen proportional area as a surrogate of extracellular matrix accumulation during fibrotic remodeling. Sirius Red combined with polarized light microscopy enhances detection of collagen fibers due to birefringence properties, enabling more specific visualization of collagen type I and III fibrils compared to conventional bright-field histology, while whole-section or region-restricted digital morphometry reduces field-selection bias in fibrosis assessment. Alternative quantitative approaches include second harmonic generation (SHG) and two-photon excited fluorescence microscopy, which enable label-free detection of fibrillar collagen and have been validated against histological staining and biochemica
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Collagen: Sirius Red Staining
Sirius Red or picrosirius red staining is a histochemical method for visualizing collagen-rich extracellular matrix in tissue sections, and collagen fibers are detected as red-stained structures under bright-field microscopy with enhanced birefringence under polarized light. Picrosirius red is useful for assessing total collagen organization, distribution, and fibrosis burden, but polarized color should not be interpreted as a definitive collagen type I versus type III readout because color is affected by fiber orientation, thickness, and packing.
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Connective Tissue: Masson's Trichrome/Collagen Trichrome Staining
Masson’s Trichrome (collagen/trichrome staining) is a histological technique that differentially stains tissue compartments using sequential acidic dyes to distinguish collagen from muscle and cytoplasmic components based on dye affinity and tissue permeability differences, enabling visualization of fibrosis and connective tissue architecture in histological sections. The classical formulation typically uses Weigert's iron hematoxylin for nuclear staining, Biebrich scarlet-acid fuchsin for cytoplasm and muscle, and aniline blue (or light green variants) for collagen, producing a characteristic blue/green collagen signal contrasted against red cytoplasm and dark nuclei. The staining principle relies on selective displacement of smaller dye molecules by larger anionic dyes in collagen-rich regions under controlled acidified conditions, which enhances collagen-specific dye retention. This property makes the method widely used for fibrosis assessment in organs such as heart, liver, lung, a
Purity & Documentation
References
Calculators
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)