AR/androgen receptor

ARV-766 is a proteolysis targeting chimera (PROTAC®) that induces a protein-protein interaction between the AR and specific E3 ubiquitin ligase complexes, leading to the ubiquitination of AR and its subsequent degradation via the proteasome. In vitro, ARV-766 degrades AR in various prostate cancer cell lines, including those harboring resistance-conferring, clinically relevant point mutations, with a half-maximal degradation concentration (DC50) of <1 nM in wild type VCaP. Importantly ARV-766 also maintains potency against the AR L702H mutant, which has been associated with resistance to some AR antagonists[2].

In vivo, ARV-766 is orally bioavailable and robustly degrades AR with a >90% observed maximum degradation (Dmax) at efficacious doses. ARV-766 significantly and dose-dependently inhibits tumor growth in murine LNCaP and VCaP xenograft models, including an enzalutamide-insensitive non-castrated VCaP model. These preclinical data supported the clinical development of ARV-766 for the treatment of men with metastatic CRPC. Selected pre-clinical data along with the chemical structure of ARV-766 will be presented[2].

ARV-766 is an orally bioavailable PROTAC® protein degrader that targets the androgen receptor (AR) and is currently being developed for the treatment of prostate cancer in a phase 2 clinical trial. In vitro studies were conducted to assess the potential of ARV-766 to cause cytochrome P450 (CYP) and transporter-mediated drug-drug interactions (DDI). The induction potential of ARV-766 on CYP enzymes was assessed in cryopreserved human hepatocytes from three donors. Following treatment for 48 hours, mRNA levels for CYP1A2, 2B6, 2C8, 2C9, 2C19, and 3A4 were determined by semiquantitative real-time polymerase chain reaction (PCR). The potential for ARV-766 to cause direct and time-dependent inhibition of the activities of CYP1A2, 2B6, 2C8, 2C9, 2C19, 2D6, and 3A was evaluated in pooled human liver microsomes (HLM). In addition, the inhibition potential against efflux transporters (Pgp and BCRP) and uptake transporters (MATE1, MATE2-K, OATP1B1, OATP1B3, OAT1, OAT3, and OCT2) was assessed using single transporter over-expressed cell monolayers (MDCK II or HEK293) or inside-out vesicles. DDI potential of ARV-766 as a victim via CYP mediated pathways was examined in HLM, human hepatocyte suspensions and recombinant CYP enzymes. ARV-766 as a substrate of Pgp and BCRP as well as OATP1B1 and 1B3 were assessed in Caco-2 and single transporter over-expressed HEK293 cells, respectively. The results showed that ARV-766 at concentrations ranging from 0.03 - 30 μM did not induce mRNA of CYP1A2, 2B6 and 2C9. Induction of CYP3A4 and CYP2C8 mRNA was observed with a maximal 2.5-8.8-fold (4-6% of positive control response) for CYP3A4 across all three hepatocyte lots, and a maximal 3.1-fold (28% of positive control response) for CYP2C8 in one of three lots. No direct or time-dependent inhibition was observed for any of the CYP isoforms after incubating HLM with ARV-766 at concentrations of 0.2-15 μM. ARV-766 did not inhibit any of the uptake transporters up to 3.75 μM tested except for MATE1 with IC50 value of 3.05 μM. ARV-766 inhibited BCRP with IC50 values of 0.21 μM (vesicle assay) and 1.55 μM (monolayer assay), and Pgp with IC50 values of 0.23 μM (vesicle assay) and >5 μM (monolayer assay). ARV-766 was relatively stable in HLM (up to 60 min), human hepatocyte suspension (up to 240 min) and major recombinant CYPs (up to 25 min) except for a 23% loss of parent with recombinant CYP3A5. Metabolite profiling using human plasma, HLM and human hepatocytes indicated that hydrolysis was the major metabolic pathway. Other minor pathways included oxidation, de-alkylation, and demethylation, which combined represent <2% of total abundance. ARV-766 exhibited low permeability in Caco-2 cell monolayers and the involvement of ARV-766 in an active efflux process was not reliably determined. In addition, ARV-766 was not likely a substrate for OATP1B1/3. These data demonstrate that ARV-766 has a low potential to cause significant DDI as an inhibitor or substrate of CYP enzymes. Clinical DDI studies with CYP3A inhibitors and inducers, and Pgp and BCRP substrates are being investigated. The 4β-hydroxy-cholesterol, a biomarker of CYP3A, is also being measured in the ongoing phase 2 clinical trial[1].

[1]. Snyder L, et al. In vitro evaluation of PROTAC® degrader ARV-766 for cytochrome P450-and transporter-mediated drug-drug interaction. Drug Metabolism and Pharmacokinetics, 2024, 55: 100881.
[2]. Snyder L, et al. Abstract ND03: Discovery of ARV-766, an androgen receptor degrading PROTAC® for the treatment of men with metastatic castration resistant prostate cancer. Cancer Research, 2023, 83(7_Supplement): ND03-ND03.
[3]. Petrylak D P, et al. A phase 2 expansion study of ARV-766, a PROTAC androgen receptor (AR) degrader, in metastatic castration-resistant prostate cancer (mCRPC). 2023.https://ascopubs.org/doi/abs/10.1200/JCO.2023.41.6_suppl.TPS290

C45H49N5O5

739.901071310043

739.373

C, 66.90; H, 6.74; F, 2.35; N, 12.14; O, 11.88

2504913-62-4

2750830-09-0 (ARV-766)

155172446

Typically exists as solid at room temperature

5.9

2

8

8

55

1330

3

O1C2C=C(C=CC=2[C@H](C2C=CC(=CC=2)N2CCC(CCN3CCN(C4C=CC5C(N([C@@H]6C(NC(CC6)=O)=O)CC=5C=4)=O)CC3)CC2)[C@H](C2C=CC=CC=2)C1)O

LXOSOQBOIBGDLD-RNMNAOLMSA-N

InChI=1S/C45H49N5O5/c51-36-11-13-38-41(27-36)55-29-39(31-4-2-1-3-5-31)43(38)32-6-8-34(9-7-32)48-20-17-30(18-21-48)16-19-47-22-24-49(25-23-47)35-10-12-37-33(26-35)28-50(45(37)54)40-14-15-42(52)46-44(40)53/h1-13,26-27,30,39-40,43,51H,14-25,28-29H2,(H,46,52,53)/t39-,40-,43-/m0/s1

(3S)-3-[6-[4-[2-[1-[4-[(3R,4S)-7-hydroxy-3-phenyl-3,4-dihydro-2H-chromen-4-yl]phenyl]piperidin-4-yl]ethyl]piperazin-1-yl]-3-oxo-1H-isoindol-2-yl]piperidine-2,6-dione

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*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.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)