Iberdomide (also known as CC-220) is a novel and potent cereblon (CRBN) modulator with an IC50 of 60 nM and can be potentially used for the treatment of systemic lupus erythematosus. Iberdomide significantly reduced Ikaros and Aiolos protein levels in B cells, T cells and monocytes. In SLE PBMC cultures, iberdomide inhibited anti-dsDNA and anti-phospholipid autoantibody production (IC50 ≈10 nM).

Physicochemical Properties

Molecular Formula	C25H27N3O5
Molecular Weight	449.4990
Exact Mass	449.195
Elemental Analysis	449.500
CAS #	1323403-33-3
Related CAS #	1560678-63-8 (HCl);1323403-33-3;
PubChem CID	67335295
Appearance	White to off-white solid powder
Density	1.3±0.1 g/cm3
Boiling Point	717.8±60.0 °C at 760 mmHg
Flash Point	387.9±32.9 °C
Vapour Pressure	0.0±2.3 mmHg at 25°C
Index of Refraction	1.629
LogP	1.07
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	6
Rotatable Bond Count	6
Heavy Atom Count	33
Complexity	732
Defined Atom Stereocenter Count	1
SMILES	O(C([H])([H])C1C([H])=C([H])C(=C([H])C=1[H])C([H])([H])N1C([H])([H])C([H])([H])OC([H])([H])C1([H])[H])C1=C([H])C([H])=C([H])C2C(N(C([H])([H])C=21)[C@]1([H])C(N([H])C(C([H])([H])C1([H])[H])=O)=O)=O
InChi Key	IXZOHGPZAQLIBH-NRFANRHFSA-N
InChi Code	InChI=1S/C25H27N3O5/c29-23-9-8-21(24(30)26-23)28-15-20-19(25(28)31)2-1-3-22(20)33-16-18-6-4-17(5-7-18)14-27-10-12-32-13-11-27/h1-7,21H,8-16H2,(H,26,29,30)/t21-/m0/s1
Chemical Name	(S)-3-(4-((4-(morpholinomethyl)benzyl)oxy)-1-oxoisoindolin-2-yl)piperidine-2,6-dione
Synonyms	CC-220; CC220; CC 220.
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
Shipping Condition	Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Biological Activity

Targets	Iberdomide targets cereblon (CRBN), a component of the CRL4CRBN E3 ubiquitin ligase complex. The cereblon-binding affinity IC50 of iberdomide is approximately 150 nM. [1]
ln Vitro	A panel of multiple myeloma (MM) cell lines (EJM, H929, KMS11, KMS128M, KMS12PE, MM1.S, MM1.R, RPM-8226, U266 cells) showed antiproliferative effects across a range of concentrations when iberdomide (CC-220; 0.01, 0.1, 1, 10 μM; 72-96 hrs) was administered[1]. In all MM cell lines, iberdomide (0.1 μM; 96 hrs) causes apoptosis[1]. On H929 cells, imiberdomide (0.1 μM; 24, 48, and 72 hours) causes time-dependent increases in the G0/G1 and sub-G1 cell cycle fractions[1]. In the KMS12BM line, iberdomide causes rapid Aiolos depletion[1]. In two of the Pomalidomide-resistant (PR) lines with cereblon mutations (EJM/PR and H929/PR), imiberdomide (0.1 μM) exhibits some anti-proliferative activity in addition to decreased levels of cereblon protein[1]. Both parental MM1.S cells and MM1.S/PR cells are equally susceptible to PBMC-mediated killing when exposed to iberdomide (0.1-1000 nM) for 72 hours[1]. Iberdomide exhibited greater antiproliferative activity than pomalidomide and lenalidomide in a panel of multiple myeloma (MM) cell lines. [1] In H929 cells (lenalidomide-sensitive) and an acquired lenalidomide-resistant (H929/LR) cell line, the antiproliferative activity order was iberdomide > pomalidomide > lenalidomide. [1] Treatment with iberdomide induced time-dependent increases in G0/G1 and sub-G1 cell cycle fractions in H929 cells. [1] Iberdomide induced greater apoptosis than pomalidomide in all tested MM cell lines at a tenfold lower concentration. [1] Treatment with 0.1 µM iberdomide led to a faster decrease in the protein levels of Ikaros, Aiolos, and c-Myc/IRF4 compared to treatment with 1 µM pomalidomide. [1] In combination with bortezomib, iberdomide showed synergistic antiproliferative effects in MM1.S cells. [1] The combination of iberdomide and bortezomib increased the apoptotic fraction to 95% compared to 87% with pomalidomide/bortezomib in MM1.S cells. [1] Iberdomide combined with daratumumab exhibited greater inhibitory effects on H929 cells than either drug alone in the presence of human-derived complement. [1] In an ADCC assay, iberdomide enhanced daratumumab-mediated antibody-dependent cellular cytotoxicity, presumably by stimulating NK cells. [1] Iberdomide induced PBMC-mediated killing of both parental MM1.5 cells and pomalidomide-resistant MM1.5/PR cells in co-culture experiments. [1] In pomalidomide-resistant (PR) cell lines with reduced but detectable cereblon levels, iberdomide showed modest antiproliferative activity. However, little to no activity was observed in PR lines with very low cereblon (MM1.R/PR, MM1.5/PR, DF15/PR). [1] In the KMS12BM/PR cell line, only iberdomide (not pomalidomide) was effective at inducing rapid depletion of Aiolos. [1] The combination of iberdomide and daratumumab showed a more pronounced dose-dependent inhibitory effect on H929/PR cells than either drug alone in complement-dependent cytotoxicity assays. [1] In KMS12PE/PR cells, the combination of iberdomide and a low concentration of bortezomib (0.25 nM) demonstrated an enhanced antiproliferative effect. [1]
ln Vivo	Higher hCRBN expression in hC343 splenocytes following 6 or 24 hours of iberdomide (CC-220; 10 mg/kg; oral gavage) is correlated with deeper IKZF1/3 downregulation in WT (C57BL/6), hC123 or-343 (representing two distinct transgenic founder lines expressing hCRBN), and mCrbn-/- mice[2].
Enzyme Assay	DMSO is used to dissolve iberdomide. The assay uses 20 mM HEPES pH 7, 150 mM NaCl, 0.005% Tween-20 assay buffer, 60 nM 6Xhis-tagged CRBN-DDB1, 30 nM cy5-conjugated cereblon modulator, and 3 nM LanthaScreen Eu-anti-His Tag antibody. The ratio of FRET to non-FRET emission is used to calculate FRET efficiency. FRET is observed by exciting at 340 nm and monitoring emission at 615 nm and 665 nm. Prior to scanning, the DMSO carrier or the competing cereblon modulating compound (iberdomide) is titrated and incubated for ten minutes[1].
Cell Assay	For antiproliferative assays, MM cell lines were treated with iberdomide across a range of concentrations, and proliferation was measured using 3H-thymidine incorporation. [1] Apoptosis was assessed by flow cytometry using Annexin-V and To-Pro-3 staining. Cells were treated with iberdomide, stained, and analyzed for apoptotic fractions. [1] Western blot analysis was performed to evaluate protein degradation. Cells were treated with iberdomide, lysed, and proteins were separated by SDS-PAGE, transferred to membranes, and probed with antibodies against targets such as Ikaros, Aiolos, ZFP91, c-Myc, and IRF4. Actin was used as a loading control. [1] For PBMC co-culture killing assays, isolated and CD3-stimulated PBMCs were incubated with iberdomide for 72 hours. These PBMCs were then co-cultured with CFSE-stained MM target cells (parental or resistant) for 4 hours. Target cell apoptosis was evaluated by flow cytometry using Annexin-V and To-Pro-3 staining. [1] Complement-dependent cytotoxicity (CDC) assays were performed by incubating target cells with iberdomide and daratumumab in the presence of human-derived complement, followed by measurement of cell viability. [1] Antibody-dependent cellular cytotoxicity (ADCC) assays were conducted. Effector PBMCs were pre-treated with iberdomide and/or daratumumab, then co-cultured with target MM cells. Target cell killing was measured. [1] Immunohistochemistry (IHC) was used on bone marrow aspirate clots from RRMM patients. Samples were stained for CD138 and cereblon, or for substrates like Ikaros, Aiolos, and ZFP91. H-scores were calculated to quantify protein expression levels. [1]
References	[1]. Leukemia. 2020 Apr;34(4):1197-1201. [2]. Blood Cancer Discov. 2021 Jul;2(4):354-369. [3]. J Med Chem. 2017 Apr 20.
Additional Infomation	Iberdomide (CC-220) has been used in trials studying the treatment of Systemic Lupus Erythematosus. Iberdomide is a modulator of the E3 ubiquitin ligase complex containing cereblon (CRL4-CRBN E3 ubiquitin ligase), with immunomodulating and pro-apoptotic activities. Upon administration, iberdomide specifically binds to the cereblon (CRBN) part of the ligase complex, thereby affecting the ubiquitin E3 ligase activity, and targeting certain substrate proteins for ubiquitination. This induces the proteasome-mediated degradation of certain transcription factors, including Ikaros (IKZF1) and Aiolos (IKZF3) which are transcriptional repressors in T-cells. This leads to a reduction of their protein levels, and the modulation of the immune system, including activation of T-lymphocytes. In addition, this leads to a downregulation of other proteins, including interferon regulatory factor 4 (IRF4), which plays a key role in the proliferation of certain cancer cell types. CRBN, the substrate recognition component of the E3 ubiquitin ligase complex, plays a key role in the ubiquitination of certain proteins. Drug Indication Treatment of mature B-cell neoplasms Iberdomide is described as a next-generation cereblon E3 ligase modulator (CELMoD). [1] It mediates its anti-myeloma activity via cereblon, leading to the targeted degradation of substrates such as Ikaros and Aiolos, and downregulation of c-Myc and IRF4. [1] The faster substrate degradation by iberdomide compared to pomalidomide may be due to its higher cereblon-binding affinity and/or improved processivity of the E3 ligase complex. [1] A phase 1b/2a clinical trial (NCT02773030) is mentioned as ongoing to determine the maximum tolerated dose of iberdomide alone or in combination with dexamethasone in relapsed/refractory multiple myeloma (RRMM) patients. [1] Pharmacodynamic analysis in patient bone marrow samples showed that iberdomide treatment significantly decreased the protein levels of Ikaros, Aiolos, and ZFP91. [1] Cereblon protein expression was highly variable in bone marrow samples from RRMM patients previously treated with IMiDs. [1]

Solubility Data

Solubility (In Vitro)

DMSO : 90~125 mg/mL ( 200.22~278.09 mM ) Ethanol : ~5 mg/mL

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.08 mg/mL (4.63 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 20.8 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: ≥ 2.08 mg/mL (4.63 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 20.8 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. Solubility in Formulation 3: ≥ 2.08 mg/mL (4.63 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.2247 mL	11.1235 mL	22.2469 mL
	5 mM	0.4449 mL	2.2247 mL	4.4494 mL
	10 mM	0.2225 mL	1.1123 mL	2.2247 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.