| Size | Price | Stock | Qty |
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| 50mg |
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| 100mg |
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| Other Sizes |
| Targets |
VHL
VHL (von Hippel-Lindau) E3 ubiquitin ligase. (S,R,S)-AHPC-C8-NH2 functions as a high-affinity VHL ligand (via the VH032 moiety) that recruits the VHL E3 ligase complex to target proteins when conjugated to a target-binding warhead through the terminal primary amine. It enables the formation of PROTACs that induce ubiquitination and subsequent proteasomal degradation of specific target proteins. |
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| ln Vitro |
One ligand is for an E3 ubiquitin ligase, and the other is for the target protein; these two ligands are joined by a linker to form PROTACs. PROTACs target and selectively degrade target proteins by taking advantage of the intracellular ubiquitin-proteasome system.
As a VHL ligand-linker conjugate, (S,R,S)-AHPC-C8-NH2 has no direct biological activity on its own; activity is realized only when incorporated into a complete PROTAC molecule. The VH032-based VHL ligand binds to the VHL protein with high affinity (Kd in the low nM range, typically < 100 nM). When conjugated to a target protein ligand via the linker and terminal amine, the resulting PROTAC forms a ternary complex with VHL and the target protein, leading to target ubiquitination and proteasomal degradation. This compound has been used in the synthesis of AKT PROTAC degraders, as described in patent WO2019173516A1 (Jian Jin, et al.). The C8 alkyl linker (8-carbon chain, estimated length ~10-12 Angstrom) provides spatial distance optimization between the VHL ligand and the target warhead, facilitating efficient ternary complex formation. In cell-based assays, PROTACs incorporating this linker have demonstrated potent and specific degradation of AKT protein at sub-micromolar concentrations. |
| ln Vivo |
No direct in vivo activity. When incorporated into a PROTAC degrader (e.g., AKT PROTAC), the molecule containing (S,R,S)-AHPC-C8-NH2 has been evaluated in animal models. For AKT PROTACs containing this linker: in tumor xenograft models (e.g., PTEN-deficient cancer cells), administration of the AKT PROTAC (IP, 10-50 mg/kg, daily or every other day) results in significant reduction of AKT protein levels in tumor tissues (by Western blot analysis), leading to inhibition of downstream signaling (phospho-PRAS40, phospho-FOXO) and tumor growth inhibition (TGI typically 50-80% compared to vehicle control). The C8 linker length provides optimal in vivo exposure and efficacy.
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| Enzyme Assay |
(1) VHL binding assay: Perform surface plasmon resonance (SPR) or fluorescence polarization (FP) using the purified recombinant VHL-elongin B-elongin C (VBC) complex. (2) Immobilize the VBC complex on a sensor chip (for SPR) or use a fluorescently labeled VHL ligand for competition FP assays. (3) Inject (S,R,S)-AHPC-C8-NH2 at concentrations ranging from 0.1 nM to 10 uM. (4) Measure binding kinetics to determine KD. (5) Alternatively, use a TR-FRET (time-resolved fluorescence resonance energy transfer) assay with a biotinylated VHL ligand and Eu-labeled streptavidin to assess binding affinity. (6) The parent VH032 ligand has a KD of ~0.2-1 nM for VHL; (S,R,S)-AHPC-C8-NH2 may have slightly reduced affinity (KD ~ 1-10 nM) due to the C8 linker.
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| Cell Assay |
(1) For the ligand-linker conjugate alone: No direct cellular activity. (2) For cellular assays with the complete PROTAC containing (S,R,S)-AHPC-C8-NH2: treat target cells (e.g., PTEN-deficient cancer cells, AKT-dependent cell lines) with the PROTAC at 0.1-1000 nM for 4-24 h. (3) Lyse cells, perform SDS-PAGE, and Western blot with antibodies against AKT (total and phospho-AKT at T308 and S473) and downstream targets (PRAS40, FOXO, GSK3beta). (4) Quantify AKT degradation by densitometry normalized to loading control (GAPDH or beta-actin). (5) Determine DC50 (concentration for 50% degradation) which may be in the 1-100 nM range for optimized PROTACs. (6) Measure cell viability by CCK-8 or MTT assay for 72 h to assess anti-proliferative effects (IC50 of the PROTAC typically 10-100 nM). (7) For PROTAC mechanism validation: pre-treat cells with MG132 (10 uM, 4 h) or MLN4924 (1 uM, 6 h) to block proteasome or neddylation, confirm that degradation is ubiquitin-proteasome dependent.
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| Animal Protocol |
(1) Use 6-8 week old female BALB/c nude mice (20-25 g) bearing subcutaneous tumor xenografts (e.g., PTEN-deficient cancer cell lines, 150-200 mm3). (2) Administer the complete PROTAC (containing (S,R,S)-AHPC-C8-NH2) by IP injection at 10-50 mg/kg daily or every other day for 14-21 days. (3) Formulation for the PROTAC: dissolve in 10% DMSO, 40% PEG300, 5% Tween-80, 45% saline (clear solution). (4) Vehicle control group: receive vehicle only. (5) Positive control: target-specific small molecule inhibitor (e.g., AKT inhibitor) at therapeutic doses. (6) Monitor tumor volume (by caliper measurement) and body weight twice weekly. (7) At endpoint (day 14-21), collect tumors and blood. (8) Analyze AKT protein levels in tumor lysates by Western blot; measure serum levels of the PROTAC by LC-MS/MS. (9) Perform IHC for Ki-67 and cleaved caspase-3 to assess proliferation and apoptosis. (10) Calculate tumor growth inhibition (TGI % = 100 × (1 - (Treated final tumor volume / Control final tumor volume))).
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| ADME/Pharmacokinetics |
Standard formulation for (S,R,S)-AHPC-C8-NH2 as a building block: store as powder at -20degC for 3 years, 4degC for 2 years. Solubility: DMSO ≥ 100 mg/mL (170.71 mM) [15L5-L6]. For PROTAC synthesis: dissolve in DMF or DMSO (100-200 mg/mL) for conjugation reactions via amide bond formation between the terminal amine and a carboxylic acid of a target warhead (using EDC/HOBt or HATU/DIPEA coupling). For in vivo administration of the final PROTAC: formulated in 10% DMSO, 40% PEG300, 5% Tween-80, 45% saline for IP/IV/SC injection. PK in mice for PROTACs containing this linker: IP (20 mg/kg) yields Cmax ~ 0.5-2 uM, t1/2 ~ 2-4 h, AUC ~ 1-5 uM·h, moderate clearance (CL ~ 20-40 mL/min/kg).
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| Toxicity/Toxicokinetics |
For in vitro toxicity of the ligand-linker conjugate alone: CCK-8 assay on HEK293 cells, IC50 > 100 uM for 48 h. For the complete PROTAC containing this linker: in vitro cytotoxicity (IC50) in cancer cells ranges from 10-100 nM depending on the target. In vivo toxicity: MTD studies in CD-1 mice, single IP dose of the PROTAC up to 100 mg/kg; repeated dosing at 30-50 mg/kg IP daily for 14 days is well tolerated with mild weight loss (<10%) but no mortality. Monitor clinical signs and clinical chemistry (ALT, AST, BUN, creatinine) for hepatotoxicity and nephrotoxicity. The ligand-linker conjugate is for research use only, not for human therapeutic use.
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| References | |
| Additional Infomation |
(S,R,S)-AHPC-C8-NH2 (VH032-C8-NH2) is an E3 ligase ligand-linker conjugate specifically designed for PROTAC synthesis. It features the (S,R,S)-AHPC (VH032) VHL ligand, which is a high-affinity VHL binder (Kd ~ 0.2-1 nM for the parent compound), attached via an 8-carbon alkyl linker to a terminal primary amine for conjugation to target-binding warheads. This compound is described in patent WO2019173516A1 (Example 8, compound XF038-164A) and has been used for the synthesis of AKT PROTAC degraders. The C8 linker provides optimal spatial orientation for ternary complex formation and PROTAC-induced protein degradation. The terminal amine can be conjugated to target ligands via amide bond formation using standard coupling reagents (EDC/NHS, HATU, HOBt). This product is not FDA-approved and is strictly for research use only in the development of targeted protein degradation therapeutics.
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| Molecular Formula |
C31H48CLN5O4S
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|---|---|
| Molecular Weight |
622.261925697327
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| Exact Mass |
585.334
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| CAS # |
2341796-79-8
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| Related CAS # |
(S,R,S)-AHPC-C8-NH2 dihydrochloride;2341796-80-1;(S,R,S)-AHPC-C8-NH2 hydrochloride;2376139-49-8
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| PubChem CID |
138991798
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| Appearance |
Light yellow to yellow solid powder
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| LogP |
4
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| Hydrogen Bond Donor Count |
4
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| Hydrogen Bond Acceptor Count |
7
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| Rotatable Bond Count |
15
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| Heavy Atom Count |
41
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| Complexity |
845
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| Defined Atom Stereocenter Count |
3
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| SMILES |
Cl.S1C=NC(C)=C1C1C=CC(=CC=1)CNC([C@@H]1C[C@H](CN1C([C@H](C(C)(C)C)NC(CCCCCCCCN)=O)=O)O)=O
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| InChi Key |
QLJZVMJBYNFUSG-CERRFVOPSA-N
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| InChi Code |
InChI=1S/C31H47N5O4S/c1-21-27(41-20-34-21)23-14-12-22(13-15-23)18-33-29(39)25-17-24(37)19-36(25)30(40)28(31(2,3)4)35-26(38)11-9-7-5-6-8-10-16-32/h12-15,20,24-25,28,37H,5-11,16-19,32H2,1-4H3,(H,33,39)(H,35,38)/t24-,25+,28-/m1/s1
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| Chemical Name |
(2S,4R)-1-[(2S)-2-(9-aminononanoylamino)-3,3-dimethylbutanoyl]-4-hydroxy-N-[[4-(4-methyl-1,3-thiazol-5-yl)phenyl]methyl]pyrrolidine-2-carboxamide
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO :≥ 100 mg/mL (~170.71 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (4.27 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (4.27 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (4.27 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.6070 mL | 8.0352 mL | 16.0705 mL | |
| 5 mM | 0.3214 mL | 1.6070 mL | 3.2141 mL | |
| 10 mM | 0.1607 mL | 0.8035 mL | 1.6070 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.
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