CC-90009: A Cereblon E3 Ligase Modulating Drug That Promotes Selective Degradation of GSPT1 for the Treatment of Acute Myeloid Leukemia

Joshua D. Hansen,* Matthew Correa, Matt Alexander, Mark Nagy, Dehua Huang, John Sapienza,

Drug Annotation

CC-90009: A Cereblon E3 Ligase Modulating Drug That Promotes

Selective Degradation of GSPT1 for the Treatment of Acute Myeloid

Leukemia

Gang Lu, Laurie A. LeBrun, Brian E. Cathers, Weihong Zhang, Yang Tang, Massimo Ammirante,

Rama K. Narla, Joseph R. Piccotti, Michael Pourdehnad, and Antonia Lopez-Girona

Cite This: J. Med. Chem. 2021, 64, 1835 −1843

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sı Supporting Information

ABSTRACT: Acute myeloid leukemia (AML) is marked by signiﬁcant unmet

clinical need due to both poor survival and high relapse rates where long-term

disease control for most patients with relapsed or refractory AML remain dismal.

Inspired to bring novel therapeutic options to these patients, we envisioned

protein degradation as a potential therapeutic approach for the treatment of

AML. Following this course, we discovered and pioneered a novel mechanism of

action which culminated in the discovery of CC-90009. CC-90009 represents a

novel protein degrader and the ﬁrst cereblon E3 ligase modulating drug to enter

clinical development that speciﬁcally targets GSPT1 (G1 to S phase transition 1)

for proteasomal degradation. This manuscript brieﬂy summarizes the mechanism

of action, scientiﬁc rationale, medicinal chemistry, pharmacokinetic properties,

and eﬃcacy data for CC-90009, which is currently in phase 1 clinical

development.

INTRODUCTION

induction/consolidation treatment, resulting in 5-year overall

survival (OS) of 40% for patients younger than 60 years. AML,

however, is mostly a disease of older individuals with a median

age at diagnosis of 68. Of those who are 65 years or older, up

■

CC-90009 2-(4-chlorophenyl)-N-((2-(2,6-dioxopiperidin-3-

yl)-1-oxoisoindolin-5-yl)methyl)-2,2-diﬂuoroacetamide, is a

cereblon E3 ligase modulating drug (CELMoD) with

antiproliferative and proapoptotic activity against acute

myeloid leukemia (AML). CC-90009 derives its anti-AML

eﬃcacy through degradation of GSPT1 (G1 to S phase

transition 1) and is currently in phase 1 clinical trials for

relapsed/refractory acute myeloid leukemia.

to 70% will not survive beyond 1 year. The recent FDA

approval of the combination therapy venetoclax plus

azacitidine has established a new standard of care for older

patients with newly diagnosed AML by demonstrating a

median overall survival of 14.7 months. This approval also

provides an opportunity to combine with additional novel

agents to improve the survival beneﬁt further. Nonetheless,

for patients who are refractory or relapse from frontline

therapy, the prognosis remains dire.

Protein Degradation. Targeted protein degradation

represents a new paradigm in drug discovery, in which small

molecules can be used to induce novel protein−protein

interactions and enable destruction of target proteins that drive

disease. Under the canopy of protein degradation, multiple

modes of interaction have been reviewed that induce protein−

Acute myeloid leukemia is a relatively rare malignancy

(

∼20 000 estimated diagnoses domestically in 2020) and is

considered an orphan disease by the US Food and Drug

Administration.

Despite advances in understanding the

pathogenesis of AML, there remains signiﬁcant clinical need

due to both poor survival and high relapse rates in AML where

the standard of care for ﬁt patients is conventional intensive

cytotoxic chemotherapy. 7+3 chemotherapy (7 days of

cytarabine and 3 days of daunorubicin) and hematopoietic

stem cell transplantation (HSCT) form the mainstay of

Received: August 26, 2020

Published: February 16, 2021

2021 American Chemical Society

Journal of Medicinal Chemistry

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Drug Annotation

Figure 1. (A) Structures of lenalidomide, pomalidomide, and CC-885. (B) Proposed mechanism of action for CC-90009. (C) NB4 cells were

treated for 4 h and then processed for Western blotting. CC-90009 selectively degrades GSPT1 (lenalidomide and CC-885 shown for comparison),

which can be rescued with cotreatment of proteasome inhibitor MG132.

protein proximity, and more speciﬁcally, two modes of

induce recruitment followed by ubiquitination of substrate

CRBN

degraders known as heterobifunctional degraders (PRO-

proteins to the CRL4

E3 ubiquitin ligase, after which,

TACs)

and molecular glues (CELMoDs) have been

ubiquitin tagged proteins are traﬃcked to and subsequently

10−12

reviewed. Both PROTACs and CELMoDs use protein

degradation mechanisms characterized with recent advance-

ment of small molecule degraders into clinical phase

investigation.

Induction of protein degradation as a therapeutic strategy

has been clinically validated by the class of immunomodulatory

drugs developed by Celgene which include lenalidomide and

pomalidomide (Figure 1A). Removal of disease-driving

proteins translates to the therapeutic beneﬁts derived from

immunomodulatory drug treatment. Speciﬁcally, CELMoDs

degraded by the 26S proteasome.

Small molecules utilized

for this purpose are commonly referred to as “molecular glues”

for their ability to draw two unrelated proteins together

through ternary complex formation. This interaction between

the two unrelated proteins achieved through ternary complex

formation, does not occur in the absence of “molecular glue”.

While lenalidomide and pomalidomide are known to derive

eﬃcacy in multiple myeloma via degradation of the tran-

scription factors IKZF1/3, CC-90009 is a novel molecular

glue that directs an alternative degradation proﬁle in which

IKZF1/3 are spared and, instead, GSPT1/eRF3a (G1 to S

(

including immunomodulatory drugs) have the capacity to

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Journal of Medicinal Chemistry

Drug Annotation

phase transition 1/eukaryotic Release Factor 3a) is degraded

indiscriminate cell killing. This is highlighted by its cytotoxic

activity against the nontumor human epithelial cell line

(Figure 1B and C).

(

THLE-2), a line that can help prioritize compounds for

RESULTS AND DISCUSSION

■

having lower probability of causing adverse events in vivo

(Table 2).

Focused on employing protein degradation as a strategy to

address high unmet medical need, our goal was to discover a

novel CELMoD that demonstrated improved clinical eﬃcacy

and an expanded treatment scope in AML which would

potentially beneﬁt patients across all age stratiﬁcations. To

widen the possibilities of ﬁnding novel proteins that when

degraded cause anti-AML activity, we envisioned that a

phenotypic screen of our cereblon (CRBN) modulator library

would be an expedient approach. Using this type of unbiased

approach could aﬀord potential to address an incomplete

understanding and complexity found with AML, and in

parallel, uncover novel biology not yet linked to AML with a

ﬁrst in class opportunity. It should be noted that there are

disadvantages when employing phenotypic screening such as

hit validation and target deconvolution. However, in our

case, the latter could be addressed by interrogating a focused

library of CRBN-interacting chemical matter. In this manner,

by using a phenotypic screen with CRBN-directed chemical

matter, we already knew “half the target” (CRBN). Once

shown to be CRBN-dependent, we could unravel the

ubiquitylated target protein via pull down or by proteome

analysis.

Table 2. Comparison of the Antiproliferative Potency for

CC-885 against a Panel of AML Cell Lines and THLE-2

cell line

KG-1

HL-60

KG-1a

KASMUI

NB4

MV-4-11

MOLM-13

OCI-AML2

OCI-AML-3

U-937

CC-885 IC₅₀(μM)

0.007

0.003

0.008

0.010

0.002

0.005

0.008

0.021

0.073

0.011

THLE-2

Because 3 did show potent and broad activity against the

AML cell line panel, it provided a structural template for us to

explore the possibility of identifying analogues with increased

therapeutic index. To achieve this aim, we explored SAR of the

chemical template and monitored the in vitro selectivity ratio

between the various AML cell lines contained within our

phenotypic panel and the THLE-2 line. The KG-1 cell line is

highlighted in this manuscript as a representative example to

demonstrate SAR in the series. A thorough investigation to

optimize or replace the terminal 3-Cl, 4-Me aryl group was

made but without improvement of the in vitro selectivity ratio.

Additionally, attempts were made to substitute at either urea-

nitrogen which also failed to yield improved in vitro selectivity

Because AML is characterized by high heterogeneity and

coupled with our desire to broadly transform the disease

landscape, we selected a panel of cell lines designed to

represent many known genetic mutations to screen our

CELMoD compound library for breadth of activity and

develop SAR (Table 1). Using this panel of AML cell lines

Table 1. Panel of AML Cell Lines and Associated

Characterization

windows, and this approach was ultimately abandoned.

cell line

KG-1

HL60

genetic traits

AML FAB classiﬁcation

Continued empirical modiﬁcations led us to discover that the

most relevant impact on selectivity vs THLE-2 cells was made

via SAR investigations around the urea linker. To set a baseline

for comparison with the urea functionality, we utilized a

derivative of 3, the unsubstituted phenyl urea 4 which, like 3,

also exhibited a low in vitro selectivity ratio when comparison

was drawn between KG-1 and THLE-2 cells (Table 3).

FGFR1 Act; NRas

Myc^a^m^p^lⁱ^fⁱ^e^d

FAB M0/1

FAB M2

FAB M3

FAB M0/1

FAB M2

FAB M5

FAB M4

FAB M5

FAB M5a

FAB M4

NB4

Pml-Rara

KG-1a

FGFR1 act NRas

RUNX1-RUNX1T1; KIT

N822 K

Kasumi-1

MV4-11

OCI-AML-2

U937

MOLM13

OCI-AML3

ITD

Mll-Af4; FLT3

R635W

DNMT3A

CALM-AF10

Mll-Af9; FLT3

Table 3. SAR Comparison of the Antiproliferative Potency

against AML Cell Line KG-1 and THLE-2

ITD

R882C

NPM1c; DNMT3A

FAB stands for French-American-British and is a classiﬁcation of

AML into subtypes M0 to M7 based on cell origin and maturity.

for the phenotypic screen rather than rely on a single cell line

was also part of the strategy to evaluate activity trends

holistically, and allowed incorporating activity breadth (in

AML cell lines) versus toxicity as a proﬁle of interest.

Structure−Activity Relationships (SAR). Recently, we

described the SAR of a series of urea containing compounds

represented by 3 (Figure 1A) which were observed to degrade

the proteins Aiolos (IKZF3), Ikaros (IKZF1), and GSPT1

KG-1 IC₅₀THLE-2 IC

in vitro

analogue

(μM)

selectivity ratio

X = NH

0.015

0.037

>10

0.060

0.231

>10

4×

6×

X = CH₂

X = none

benzamide)

(

eRF3a) with variable levels of selectivity. Compound 3

(

displayed potent and broad activity against the AML cell line

panel utilized for our phenotypic screen. However, compound

X = CO

0.009

>10

0.196

>10

22×

^X⁼^C⁽^C^H3⁾

was considered unsuitable for further development due to

^X⁼^C^F2

0.097

>10

>100×

concerns about its potential toxicity proﬁle as a result of

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Drug Annotation

Removal of the distal nitrogen atom and replacement with

carbon led to amide 5, which had little impact on the

selectivity ratio. Truncation of the structure by deletion of the

distal urea nitrogen atom provided benzamide 6 which was

inactive. Comparison of the phenylacetamide functionality in 5

to the oxo acetamide contained in 7 gave us the ﬁrst hint that

selectivity between tumor and normal cell lines could be

tempered. Although a THLE-2 IC₅₀of 196 nM was still

concerning, compound 7 did maintain single digit nanomolar

activity against KG-1 with a 22× selectivity window against

THLE-2. The SAR in this region was found to be highly

sensitive as dimethyl substitution on the phenyl acetamide

carbon was not tolerated (8 versus 5). However, we were

pleased to discover that the smaller diﬂuoro substituent in

compound 9 maintained activity below 100 nM against KG-1

cells, and importantly, an IC₅₀> 10 μM was observed in

THLE-2 cells. This diﬂuoro substitution pattern yielded an in

vitro selectivity ratio >100× (KG-1 vs THLE-2 potency) which

provided impetus to further optimize this diﬂuoro acetamide

series and ultimately led to the selection of CC-90009 as a

clinical candidate.

In Vitro Pharmacology. To show that CC-90009-induced

loss of GSPT1 was mediated via proteasomal degradation, KG-

cells were treated with drug, which showed GSPT1

degradation by Western blot. When the KG-1 cells were

cotreated with drug plus the proteasome inhibitor bortezomib

or the neddylation inhibitor MLN4924, rescue of GSPT1

degradation was observed (Figure 2A). Loss of GSPT1 in

leukemic cells leads to activation of the integrated stress

response (ISR), which is an evolutionarily conserved

homeostatic pathway. ISR pathway activation results in the

inhibition of global protein translation and preferential

17,18

translation of the ISR eﬀector ATF4.

Thus, CC-90009

induced degradation of GSPT1 ultimately leads to accumu-

lation of ATF4 plus its transcriptional targets CHOP and

ATF3 with subsequent induction of apoptosis in leukemic

19,20

blasts (Figure 2B).

CC-90009 is a potent GSPT1 degrader with a degradation

EC₅₀= 9 nM and Y_m_i_n= 49 or in other terms with a

degradation maximum (D_m_a_x) = 51% when measured in a

cellular degradation assay at 4 h. When GSPT1 degradation

was measured at 20 h, the EC₅₀remained the same, but the

depth of degradation increased (Y_m_i_n= 12; D_m_a_x= 88). CC-

Figure 2. (A) Immunoblot analysis of KG1 cells treated with DMSO

or CC-90009 for 4 h. Where indicated, cells were pretreated with

bortezomib or MLN4924 for 30 min. (B) Immunoblot analysis of

whole cell extracts of KG-1 cells. Cells were incubated with DMSO or

0009 is selective in its degradation over other proteins

(

IKZF1/3 EC₅₀> 10 μM). Further assessment of CC-90009’s

00 nM CC-90009 and lysed at the indicated time points. Arrows

selectivity by proteomic experiments with concentration range

from 0.20 μM up to 3 μM with 4 or 6 h incubation shows

GSPT1 is the only detectable protein degraded with greater

pointing to bands in the blot on the right designate the cleaved forms

of caspase-3.

than −1 log2 fold change (∼75% degradation).

CC-90009 demonstrated antiproliferative activity in over

0% of the human AML cancer cell lines tested, and when

xenograft model of AML. Treatment was initiated 6 weeks

postinoculation with vehicle and CC-90009. Treatment groups

were terminated either on Day 7 (following 5 consecutive days

of 5 mg/kg BID CC-90009) or on Day 11 (following 10

consecutive days of 2.5 mg/kg BID CC-90009) post treatment

initiation. The primary end point for the study was percentage

examined in primary patient AML blasts, exerted this eﬀect

through rapid induction of apoptosis. In ex vivo studies

employing AML patient bone marrow samples, growth

inhibition of leukemic cells was 2- to 5-fold greater than the

inhibition of normal adult lymphocytes. In 9 of the ﬁrst 10

patient samples examined, CC-90009 was highly active and

of hCD33 /CD45 cells in the bone marrow by FACS analysis.

Treatment with 5 mg/kg CC-90009 BIDx5 resulted in a

signiﬁcant (p = 0.0013) 54.0% reduction of human CD33+/

CD45+ tumor cells in the BM when compared to the vehicle

control group. Treatment with 2.5 mg/kg CC-90009 BIDx10

resulted in a signiﬁcant (p < 0.0001) 71.5% reduction of

had an average EC ∼ 6 nM. When an additional 30 patient

samples were evaluated, CC-90009 retained activity in ∼80%

of the samples, consistent with the in vitro AML cell lines

tested.

In Vivo Pharmacology. CC-90009 eﬀectively reduced

tumor cell content in femur bone marrow in an HL-60

human CD33 /CD45 tumor cells in the bone marrow when

compared to the vehicle control group.

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Drug Annotation

Pharmacokinetics. Mouse pharmacokinetic studies were

conducted in male CD-1 mice (n = 4). CC-90009 was dosed

IV bolus to nonfasted mice at 2 mg/kg with a dose volume of 2

mL/kg in DMA/PEG400/5% dextrose in water (15:50:35).

Samples were obtained via nonserial sampling due to the

composite sampling design, and mean PK parameters are

reported and listed in Table 4.

predicted a low human clearance of approximately 1.93 mL/

min/kg and a volume of distribution of 1.48 L/kg.

In Vitro Safety. CC-90009 was determined to be modestly

active (IC₅₀= 5.3 μM) against the human ether-a-go-go-

related gene (hERG) ion channel. In a stand-alone nonpivotal

in vivo cardiovascular safety pharmacology study in cyn-

omolgus monkeys conducted with CC-90009, there were no

CC-90009-related cardiovascular eﬀects. CC-90009 (up to 2.5

μM) did not cause inhibition or induction of cytochrome P450

(CYP) enzymes in in vitro studies except for weak inhibition of

CYP2C9 (41% at 2.5 μM) and CYP2C19 (IC = 1.53 μM). In

Table 4. Rodent and Monkey Pharmacokinetic Parameters

Following IV Dosing for CC-90009

addition, CC-90009 (up to 3 μM) did not inhibit P-gp, BCRP,

MRP2, OAT1, OAT3, OATP1B1, OATP1B3, OCT2, or

MATE2-K. CC-90009 was screened against a diverse panel of

species

mouse

dose

(mL/min/kg) V_s_s(L/kg) MRT (h)

IV dose (2

mg/kg)

67.9

1.00

0.24

55 kinases at 3 μM with no inhibition observed >20%. The

rat

IV dose (2

mg/kg)

28.3 ± 0.7

6 ± 1

2.2 ± 0.5

1.1 ± 0.1

1.3 ± 0.3

4 ± 1

selectivity of CC-90009 was explored further by evaluating its

eﬀect in binding assays for 80 diﬀerent human receptors, ion

channels, and transporters, and at a concentration of 10 μM,

CC-90009 inhibited receptor binding ≥50% at only two

receptors, M (h) and M (h).

Pharmaceutical Properties. CC-90009 appears as a white

powder, and its crystallinity was conﬁrmed by XRPD.

Diﬀerential scanning calorimetry (DSC) showed a melting

onset at 228 °C. The material used for clinical studies is an oﬀ-

white material and is prepared as a racemic mixture of the R-

and S-enantiomers as the single enantiomers demonstrated

rapid in vitro and in vivo epimerization across multiple species.

The formulation used for injection of CC-90009 is a

lyophilized powder for reconstitution with sterile water to

provide a solution that can yield a dose range from 0.6 to 20

mg.

monkey IV dose (1

mg/kg)

Rat pharmacokinetic studies were conducted in jugular vein

cannulated male CD-IGS rats (n = 3). CC-90009 was dosed IV

bolus to fed rats at 2 mg/kg with a dose volume of 2 mL/kg in

DMA/PEG400/5% dextrose in water (15:50:35). The rat IV

pharmacokinetic parameters are listed in Table 4.

A monkey PK study was conducted in male cynomolgus

monkeys (n = 3). Data from the study are shown in Table 4.

CC-90009 was dosed IV bolus to nonfasted monkeys at 1 mg/

kg with a dose volume of 1 mL/kg in DMA/ethanol/PEG400/

% dextrose in water (5:10:40:45).

CC-90009 has a moderate to high clearance in the mouse,

but a moderate to low clearance in rat and cynomolgus

monkey, the preferred toxicology species. The metabolites of

nonradiolabeled CC-90009 were investigated in hepatocytes

from mouse, rat, monkey, and human. Additional metabolites

were examined in vivo in the rat. Taken together, there were

no unique human metabolites detected. Allometric scaling

Clinical Data. During the treatment period, CC-90009 is

administered intravenously on days 1 to 5, days 1 to 7, or on

days 1 to 3 and days 8 to 10 of each 28-day cycle for up to 4

cycles. Mean terminal half-life following the last day of dosing

was estimated to be approximately 9.3 h. Upon 5 days of repeat

Scheme 1. Synthesis of CC-90009

Reagents and conditions: (a) 1 M NaOH, rt, 2 h, THF; (b) EtI, 80 °C, 1 h, DMF, 66% over 2 steps; (c) SO Cl , DCM, 0 °C, 16 h, 41%; (d) 3-

aminopiperidine-2,6-dione·HCl, triethylamine, DMF, 70 °C, 16 h, 61%; (e) NiCl , NaBH , Boc O, NMP, rt, 15 h, 37%; (f) 4 M HCl-dioxane, rt, 2

h, 93%; (g) MeOH-H SO (50-1), 65 °C, 18 h, 95%; (h) NBS, azo-isobutyronitrile, 85 °C, 18 h, ACN, 66%; (i) 3-aminopiperidine-2,6-dione.HCl,

triethylamine, rt for 25 h, 44%; (j) 1,1′-bis(diphenylphosphino)ferrocene, ZnCN, ZnOAc, Pd (dba) , 120 °C for 20 h, 57%; (k) MsOH, Pd/C,

DMA, 50 psi, 40 °C for 20 h, 40%, (l) 2-(4-chlorophenyl)-2,2-diﬂuoroacetic acid, HATU, DIEA, DMF, 16 h, 86%.

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Journal of Medicinal Chemistry

Drug Annotation

dosing, a 1.2- to 2.7-fold accumulation of total systemic

exposure was observed across dosing schedules. Across all

doses and schedules, the geometric mean total clearance was

can be employed toward further addressing unmet medical

need.

.96 L/h, and volume of distribution was 61.69 L. In general,

EXPERIMENTAL SECTION

■

the exposures increased with dose in each dosing schedule.

GSPT1 levels in CD3 T cells from individual patients treated

General. Compounds were named using ChemDraw Ultra. All

materials were obtained from commercial sources and used without

further puriﬁcation, unless otherwise noted. Chromatography solvents

were HPLC grade and used as purchased. All air-sensitive reactions

were carried out under a positive pressure of an inert nitrogen

atmosphere. Chemical shifts (δ) are reported in ppm downﬁeld of

TMS and coupling constants (J) are given in Hz. Thin Layer

Chromatography (TLC) analysis was performed on Whatman thin

layer plates. The purity of ﬁnal tested compounds was ≥95% as

determined by HPLC using the following method: gradient (5−95%

ACN + 0.075% formic acid in water +0.1% formic acid over 8 min,

followed by 95% ACN + 0.075% formic acid for 2 min); ﬂow rate 1

mL/min, column Phenomenex Luna 5 μ PFP(2) 100A (150 mm ×

with CC-90009 on D1−5 were measured by ﬂow cytometry,

and dose-dependent decreases in GSPT1 levels were observed

in peripheral blood blasts and T cells. At a dose of 2.4 mg, 90%

GSPT1 reduction was observed.

Chemistry. The synthesis of CC-90009 (16) is outlined in

Scheme 1. CC-90009 is made by coupling 2-(4-chlorophenyl)-

,2-diﬂuoroacetic acid with the key intermediate 13. The key

benzylamine 13 can be derived in multiple ways, one of which

begins with Fischer esteriﬁcation of 4-bromo-2-methylbenzoic

acid 14 followed by radical bromination of the tolyl methyl

.60 mm).

group. Formation of the bicyclic lactam was accomplished

using 3-aminopiperidine-2,6-dione as the nucleophile for

bromide displacement followed by further condensation onto

the ester to form lactam 15. The requisite aminomethyl

functionality was installed via palladium-mediated cyanide

insertion to the aryl bromide with subsequent reduction to

yield 13. Alternatively, 13 could be synthesized starting from

the lactone 10 by introduction of the amine functionality via

nitrile reduction. This alternate starting point provides a route

found to be convenient when used on larger scale and avoids

the use of palladium near the end of the synthetic sequence.

Synthesis. Compound 13 has been previously described in the

26−29

literature.

-((2-(2,6-Dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)-

methyl)-3-phenylurea (4). To a stirred mixture of 3-(5-(amino-

methyl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione methanesulfonate

(0.37 g, 1.0 mmol) and phenylisocyanate (0.12 g, 1.0 mmol) in

acetonitrile (10 mL) was added triethylamine (0.28 mL, 2.0 mmol) at

room temperature under nitrogen. After 2 h, 1 N aq. HCl (10 mL)

was added, and the solids were isolated by ﬁltration and washed with

water (20 mL). The solids were triturated in ethyl acetate (10 mL) for

18 h, then isolated by ﬁltration, washed with ethyl acetate (40 mL),

and dried under vacuum to give 1-((2-(2,6-dioxopiperidin-3-yl)-1-

oxoisoindolin-5-yl)methyl)-3-phenylurea as a white solid (0.38 g, 0.97

CONCLUSION

■

mmol, 97% yield, HPLC purity >97%). H NMR (300 MHz, DMSO-

During the course of SAR investigation, we discovered that

replacement of the urea moiety in 3 with 2,2-diﬂuoroacetamide

could alter the proﬁle of degraded proteins and demonstrated

selectivity for GSPT1 over IKZF1/3. This optimized

functionality held by CC-90009 led to the desired activity

proﬁle which maintained broad anti-AML activity but

increased selectivity against a noncancer line like THLE-2,

indicating a higher probability of achieving relevant therapeutic

index. CC-90009 was shown to have an in vitro ADME and

safety proﬁle that supported additional studies in vivo and

ultimately advanced into safety and eﬃcacy studies in subjects

with AML where a dose level of 2.4 mg resulted in 90%

GSPT1 degradation.

) δ 10.97 (s, 1H), 8.61 (s, 1H), 7.69 (d, J = 7.8 Hz, 1H), 7.52 (s,

H), 7.47−7.37 (m, 3H), 7.22 (t, J = 7.8 Hz, 2H), 6.89 (t, J = 7.3 Hz,

H), 6.72 (t, J = 6.0 Hz, 1H), 5.10 (dd, J = 13.2, 5.1 Hz, 1H), 4.62−

.11 (m, 4H), 3.03−2.79 (m, 1H), 2.62−2.56 (m, 1H), 2.42−2.19

(m, 1H), 2.03−1.91 (m, 1H); MS (ESI) m/z 393.2 [M + 1] .

N-((2-(2,6-Dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)-

methyl)-2-phenylacetamide (5). To a solution of 2-phenylacetyl

chloride (200 mg, 1.29 mmol) in acetonitrile (20 mL) was added 3-

(5-(aminomethyl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione metha-

nesulfonate (434 mg, 1.17 mmol). The mixture was cooled to 0 °C,

and triethylamine (264 mg, 2.6 mmol) was added dropwise. The

reaction mixture was stirred at room temperature for 18 h. More 2-

phenylacetyl chloride (200 mg) and more triethylamine (264 mg) was

added and stirring continued for 5 h. To the mixture was added 1N

aq. HCl (20 mL), dropwise. The resulting solids were collected by

ﬁltration, washed with ethyl acetate (30 mL), and dried in vacuo to

give N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2-

The ability to design small molecules that catalytically

orchestrate removal of “undruggable” proteins has tremendous

potential to improve lives of countless patients when applied

toward disease-driving proteins. An example where controlled

protein homeostasis has revolutionized disease landscape can

be gleaned from multiple myeloma, where proteasome

inhibitors such as bortezomib and protein degraders such as

lenalidomide have become the backbone therapy and

phenylacetamide (216 mg, 0.55 mmol, 47% yield, HPLC purity

>98%). H NMR (DMSO-d ) δ 10.98 (s, 1H), 8.65 (t, J = 5.9 Hz,

1H), 7.66 (d, J = 7.7 Hz, 1H), 7.46−7.15 (m, 7H), 5.10 (dd, J = 5.1,

3.2 Hz, 1H), 4.50−4.19 (m, 4H), 3.50 (s, 2H), 3.01−2.82 (m, 1H),

.63 (br. s, 1H), 2.39 (qd, J = 4.3, 13.2 Hz, 1H), 2.09−1.87 (m, 1H).

MS (ESI) m/z 392.16 [M + 1] .

N-((2-(2,6-Dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)-

methyl)benzamide (6). To a stirred mixture of 3-(5-(amino-

methyl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione methanesulfonate

(0.50 g, 1.80 mmol) and benzoyl chloride (0.25 g, 1.80 mmol) in

acetonitrile (20 mL) was added triethylamine (0.51 mL, 3.60 mmol)

at room temperature under nitrogen. After 1 h, 1N aq. HCl (20 mL)

was added, and the mixture was stirred for 10 min. The product was

isolated by ﬁltration, washed with 1N aq. HCl (20 mL) and

acetonitrile (20 mL), and dried overnight in vacuo to give N-((2-(2,6-

dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)benzamide as a

dramatically increased ﬁve-year survival rates. While

lenalidomide was commercialized prior to understanding its

degradation mechanism, CC-90009 in sharp contrast is the

ﬁrst CRBN-mediated protein degrader to enter clinical trials

24,25

since the seminal discovery by Handa

immunomodulatory drugs.

linking CRBN to the

In diversion from the IMiDs (lenalidomide, pomalidomide,

and thalidomide) that degrade IKZF1/3, CC-90009 is the ﬁrst

small molecule molecular glue to be reported that selectively

promotes the degradation of GSPT1. Our strategy employed

phenotypic screening combined with E3-ligase directed

chemical matter which helped uncover novel biology. This

strategy may provide a roadmap for future advances we hope

white solid (0.43 g, 1.14 mmol, 64% yield, HPLC purity >99%). H

NMR (400 DMSO-d ) δ 10.99 (s, 1H), 9.16 (t, J = 5.9 Hz, 1H), 7.91

(

d, J = 7.0 Hz, 2H), 7.70 (d, J = 7.7 Hz, 1H), 7.62−7.42 (m, 5H),

5.11 (dd, J = 5.0, 13.1 Hz, 1H), 4.60 (d, J = 5.9 Hz, 2H), 4.45 (d, J =

17.4 Hz, 1H), 4.31 (d, J = 17.4 Hz, 1H), 3.05−2.79 (m, 1H), 2.68−

840

J. Med. Chem. 2021, 64, 1835−1843

Journal of Medicinal Chemistry

Drug Annotation

.53 (m, 1H), 2.47−2.27 (m, 1H), 2.06−1.93 (m, 1H); MS (ESI) m/

NMR (400 MHz, DMSO-d ) δ 11.00 (s, 1H), 9.67 (t, J = 6.25 Hz,

z 378.15 [M + 1] .

1H), 7.67 (d, J = 7.81 Hz, 1H), 7.50−7.62 (m, 5H), 7.34- 7.42 (m,

2H), 5.11 (dd, J = 5.08, 13.28 Hz, 1H), 4.38−4.47 (m, 3H), 4.24−

4.31 (m,1H), 2.86−2.97 (m, 1H), 2.55−2.64 (m, 1H), 2.32−2.45 (m,

1H), 1.99 (dtd, J = 2.34, 5.25, 12.55 Hz, 1H). MS (ESI) m/z 428.14

N-((2-(2,6-Dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl) meth-

yl)-2-oxo-2-phenylacetamide (7). Step A. To a stirred solution

of 2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-carbonitrile (10 g,

7.13 mmol) in dimethylacetamide (320 mL) were added

[M+l] .

methanesulfonic acid (2.6 mL, 40.85 mmol) and 10% dry palladium

2-(4-Chlorophenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-l-oxoisoindo-

lin-5-yl)methyl)-2,2-diﬂuoroacetamide (16). To 3-(5-(aminometh-

yl)-1 -oxoisoindolin-2- yl)piperidine-2,6-dione methanesulfonate 13

(15.0 g, 40.6 mmol) in Ν,Ν-dimethylformamide (100 mL) was added

1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]-

pyridinium 3-oxid hexaﬂuorophosphate (16.9 g, 44.7 mmol) and 2-(4-

chlorophenyl)-2,2-diﬂuoroacetic acid (8.4 g, 40.6 mmol) followed by

diisopropylethylamine (21.3 mL, 122 mmol). The mixture was

allowed to stir at 25 °C for 24 h and then partitioned between 1500

mL of water and 1500 mL of EtOAc. The organic layer was washed

with saturated sodium bicarbonate solution (2 × 500 mL), 1 N HCl

solution (2 × 500 mL), and then brine (2 × 300 mL). The organic

layer was then concentrated under reduced pressure to aﬀord a white

solid which was slurried in water for 30 min and collected by vacuum

ﬁltration. The solids were digested with isopropanol and reﬂuxed for 1

h. After cooling to room temperature, the solids were collected by

vacuum ﬁltration to aﬀord 2-(4-chlorophenyl)-N-((2-(2,6-dioxopiper-

idin-3-yl)-loxoisoindolin-5-yl)methyl)-2,2-diﬂuoroacetamide (16.1 g,

on carbon (4 g) and stirred at hydrogen pressure under 50 psi at 40

C for 20 h in a hydrogen vessel. The reaction mixture was ﬁltered

through Celite and washed with water (100 mL). The ﬁltrate was

concentrated under reduced pressure, and the resultant residue was

triturated with 1% methanol in dichloromethane to give 3-(5-

(

aminomethyl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione methane-

sulfonate (5.6 g, 15.17 mmol, 40% yield) as an oﬀ-white solid. MS

(ESI) m/z 272.0 [M − 1] .

Step B. To a stirred cold (0 °C) solution of 3-(5-(aminomethyl)-1-

oxoisoindolin-2-yl)piperidine-2,6-dione methanesulfonate (300 mg,

.813 mmol) in N,N-dimethylacetamide (10 mL) was added 2-oxo-2-

phenylacetic acid (122 mg, 0.813 mmol) followed by N,N-

diisopropylethylamine (0.5 mL, 2.43 mmol) and 1-[bis-

(

dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-

oxid hexaﬂuorophosphate (402 mg, 1.056 mmol) and stirred at

room temperature for 16 h. The reaction mixture was diluted with

water (100 mL), the solid obtained was ﬁltered, washed with diethyl

ether (20 mL), dried and puriﬁed by column chromatography (100−

34.8 mmol, 85.7% yield, HPLC purity >99%) as a white solid. H

00 silica) using 3% methanol in dichloromethane as eluent to give

NMR (500 MHz, DMSO-d6) δ ppm 10.98 (s, 1H), 9.68 (t, J = 6.15

Hz, 1H), 7.69 (d, J = 7.88 Hz, 1H), 7.58−7.66 (m, 4H), 7.33−7.44

(m, 2H), 5.11 (dd, J = 13.24, 5.04 Hz, 1H), 4.39−4.50 (m, 3H),

4.24−4.35 (m, 1H), 2.85−2.98 (m, 1H), 2.61 (dd, J = 15.29, 2.05 Hz,

1H), 2.39 (dd, J = 12.93, 4.73 Hz, 1H), 1.95−2.07 (m, 1H). MS

(55 mg, 0.135 mmol, 17% yield, HPLC purity >97%) as an oﬀ white

solid. H NMR (400 MHz, DMSO-d ) δ (ppm) 10.98 (s, 1H), 9.56

(

d, J = 6.2 Hz, 1H), 8.08−7.86 (m, 2H), 7.80−7.66 (m, 2H), 7.63−

.55 (m, 3H), 7.49 (d, J = 7.8 Hz, 1H), 5.12 (dd, J = 13.3, 5.1 Hz,

H), 4.58 (d, J = 6.0 Hz, 2H), 4.48 (d, J = 17.4 Hz, 1H), 4.33 (d, J =

7.3 Hz, 1H), 2.99−2.83 (m, 1H), 2.73 (s, 1H), 2.63 (s, 1H), 2.27 (s,

(ESI) m/z 462.2 [M+l] .

ePL Degradation Assay. DF15 multiple myeloma cells stably

expressing ePL-tagged GSPT1 were generated via lentiviral infection

with pLOC-ePL-GSPT1. Cells were dispensed into a 384-well plate

(Corning #3712) prespotted with compound. Compounds were

dispensed by an acoustic dispenser (ATS Acoustic Transfer System

from EDC Biosystems) into a 384-well in a 10 pt dose response curve

using 3-fold dilutions starting at 10 μM and going down to 0.0005 μM

in DMSO. A DMSO control is added to the assay. Twenty-ﬁve

microliters of media (RPMI-1640 + 10% Heat Inactivated FBS + 25

mM Hepes+1 mM Na Pyruvate + 1× NEAA + 0.1% Pluronic F-68 +

1× Pen Strep Glutamine) containing 5000 cells was dispensed per

H); MS (ESI) m/z 406.14 [M + 1] .

N-((2-(2,6-Dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)-

methyl)-2-methyl-2-phenylpropanamide (8). To a cooled (0

C) solution of 3-(5-(aminomethyl)-1-oxoisoindolin-2-yl)piperidine-

,6-dione hydrochloride (200 mg, 0.65 mmol) in N,N-dimethylace-

tamide (4 mL) were added 2-methyl-2-phenylpropanoic acid (106

mg, 0.65 mmol) and 1-[bis(dimethylamino)methylene]-1H-1,2,3-

triazolo[4,5-b]pyridinium 3-oxid hexaﬂuorophosphate (369 mg, 0.97

mmol) followed by diisopropylethylamine (0.34 mL, 1.94 mmol) and

stirred at room temperature for 16 h. The volatiles were removed

under reduced pressure, and the resultant residue was poured into

water (10 mL). The precipitated solid was ﬁltered and puriﬁed by

Reveleris C-18 reversed phase column chromatography (55−60%

acetonitrile/0.1% aqueous formic acid) to aﬀord N-((2-(2,6-

dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2-methyl-2-phenyl-

well. Assay plates were incubated at 37 °C with 5% CO for 4 h. After

incubation, 25 μL of the InCELL Hunter Detection Reagent Working

Solution (DiscoverX, cat #96-0002, Fremont, CA) was added to each

well and incubated at room temperature for 30 min protected from

light. After 30 min, luminescence was read on a PHERAstar

luminometer (Cary, NC).

propanamide (105 mg, 0.25 mmol, 39% yield, HPLC purity >99%) as

an oﬀ-white solid. H NMR (400 MHz, DMSO-d ) δ 10.98 (s, 1H),

To determine EC50 values for GSPT1 degradation, a four-

parameter logistic model (sigmoidal dose−response model) (FIT=

.02 (t, J = 6.0 Hz, 1H), 7.61 (d, J = 8.0 Hz, 1H), 7.33−7.21 (m, 7H),

.10 (dd, J = 14.0, 5.6 Hz, 1H), 4.48 (d, J = 16.8 Hz, 1H), 4.33 (d, J =

.6 Hz, 2H), 4.24 (d, J = 17.2 Hz, 1H), 2.96−2.87 (m, 1H), 2.62−

(A + ((B − A)/1 + ((C/x)D))), where C is the inﬂection point

(EC50), D is the correlation coeﬃcient, and A and B are the low and

high limits of the ﬁt, respectively) was used to determine the

compound’s EC50 value, which is the half-maximum eﬀective

concentration. The minimum Y is reference to the Y constant. In

the GSPT1 degradation assay, we used compound 3 as the control

with a Y constant = 0. The maximum limit is the Y max DMSO

control. All percent of control GSPT1 degradation curves were

processed and evaluated using Activity Base (IDBS). The maximum

limit is the Y max DMSO control. All percent of control GSPT1

degradation curves were processed and evaluated using Activity Base

(IDBS).

.58 (m, 1H), 2.44−2.39 (m, 1H), 2.38−1.97 (m, 1H), 1.49 (s, 6H);

MS (ESI) m/z 420.19 [M + 1] .

N-((2-(2,6-Dioxopiperidin-3-yl)- l-oxoisoindolin-5-yl)-

methyl)-2,2-. Diﬂuoro-2-phenylacetamide (9). 3-(5-(Aminometh-

yl)-1 -oxoisoindolin-2-yl)piperidine-2,6-dione, mesylic acid (0.050 g,

.135 mmol), was placed in a vial with Ν,Ν-dimethylformamide (1.0

mL), 2,2-diﬂuoro-2-phenylacetic acid (0.023 g, 0.135 mmol),

diisopropylethylamine (0.071 mL, 0.406 mmol), and 1-[bis-

(

dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-

oxide hexaﬂuorophosphate (0.057 g, 0.149 mmol). The reaction

mixture was stirred at room temperature for 18 h. The reaction

mixture was taken up in dimethyl sulfoxide and puriﬁed using reverse-

phase semi preparatory HPLC (5−100% acetonitrile + 0.1% formic

acid in water + 0.1% formic acid over 20 min). Fractions containing

desired product were combined, and volatile organics were removed

under reduced pressure to give N-((2-(2,6-dioxopiperidin-3-yl)-l-

Cell Culture. Acute myeloid leukemia cell lines KG-1, Kasumi-1,

U937, MOLM-13, HL-60, and MV-4-11 were purchased from

American Tissue Culture Collection (ATCC). NB-4, HNT-34,

OCI-AML2, and OCI-AML3 cell lines were purchased from Deutsche

Sammlung von Mikroorganismen and Zellkulturen GmbH. 293FT

CRBN−/−, MOLM-13 CRBN−/−, and OCI-AML2 CRBN−/− cell

lines were described previously. KG-1, Kasumi-1, U937, MOLM-13,

NB-4, and HNT-34 cell lines were maintained in Roswell Park

oxoisoindolin-5-yl)methyl)-2,2-diﬂuoro-2-phenylacetamide (0.039 g,

0.091 mmol, 67.4% yield, HPLC purity >99%) as a white solid. H

841

J. Med. Chem. 2021, 64, 1835−1843

Journal of Medicinal Chemistry

The Supporting Information is available free of charge at

Drug Annotation

Memorial Institute (RPMI) 1640 tissue culture medium (Invitrogen)

supplemented with 10% FBS, 1× sodium pyruvate, 1× nonessential

amino acids, 100 U/mL penicillin, and 100 μg/mL streptomycin. HL-

(IACUC). Animals were acclimatized to the animal housing

facility for a period of seven days prior to the beginning of the

experiment.

60 and MV-4-11 cell lines were maintained in Iscove’s modiﬁed

Dulbecco’s medium (IMDM; Invitrogen) supplemented with 10%

FBS, 1× sodium pyruvate, 1× nonessential amino acids, 100 U/mL

penicillin, and 100 μg/mL streptomycin. OCI-AML2 and OCI-AML3

cell lines were maintained in minimal essential medium (MEM;

Invitrogen) supplemented with 10% FBS, 1× sodium pyruvate, 1×

nonessential amino acid, 100 U/mL penicillin, and 100 μg/mL

streptomycin.

ASSOCIATED CONTENT

sı Supporting Information

https://pubs.acs.org/doi/10.1021/acs.jmedchem.0c01489 .

■

Biological data and molecular formula strings (CSV)

HPLC traces (PDF )

Cell Proliferation Assay. Human cancer cell lines cultured in the

growth medium recommended by the vendor were seeded into black

84-well plates containing DMSO or test compounds. The seeding

AUTHOR INFORMATION

Corresponding Author

■

density of each cell line was optimized to allow for cell growth in the

linear range during a 3-day culture period. To test the compound

eﬀect on cell proliferation in AML cell lines, 5000 to 10000 cells per

well in 200 μL complete culture media were seeded into black 96-well

plates containing DMSO or test compounds. After 72 h, cell

proliferation was assessed using CTG according to the manufacturer’s

instructions. The growth inhibitory IC₅₀values of CC-90009 were

determined using ActivityBase (IDBS).

Joshua D. Hansen − Bristol Myers Squibb, San Diego,

California 92121, United States; orcid.org/0000-0002-

4881-5247 ; Phone: 858-795-4910;

Email: joshua.hansen@bms.com

Authors

Cell Apoptosis Assay. The ability of CC-90009 to induce

apoptosis was assessed in selected AML cell lines at the time points

and compound concentrations indicated. For Annexin V/7AAD

readout by ﬂow cytometry, AML cell lines were plated into ﬂat

bottom 96-well plates (BD Falcon) at a seeding density of 0.1−0.3 ×

Matthew Correa − Bristol Myers Squibb, San Diego,

California 92121, United States

Matt Alexander − Bristol Myers Squibb, San Diego, California

92121, United States

Mark Nagy − Bristol Myers Squibb, San Diego, California

2121, United States

Dehua Huang − Bristol Myers Squibb, San Diego, California

2121, United States

106 cells per mL in 200 μL complete media. CC-90009 was dispensed

onto the plates, and the cells were incubated for 24−48 h. At the end

of the incubation period, 100 μL of cells were transferred into a 96-

well U-bottom plate (BD Falcon), centrifuged at 1200 rpm for 5 min,

and the media was removed. Then, 2.5 μL of Annexin V-AF647

John Sapienza − Bristol Myers Squibb, San Diego, California

92121, United States

(

Biolegend) and 5 μL 7AAD (Biolegend) were diluted into 100 μL of

× Annexin binding buﬀer (BD Biosciences). Fifteen minutes after

Gang Lu − Bristol Myers Squibb, San Diego, California

addition of 100 μL of Annexin V/7AAD buﬀer into each well, cells

were analyzed using the Attune Flow Cytometer (Invitrogen). The

apoptosis induction curve was processed and graphed using GraphPad

Prism Version 7 (P < 0.05, unpaired two-sided t test, is considered as

signiﬁcant).

92121, United States

Laurie A. LeBrun − Bristol Myers Squibb, San Diego,

California 92121, United States

Brian E. Cathers − Bristol Myers Squibb, San Diego,

California 92121, United States

Weihong Zhang − Bristol Myers Squibb, Summit, New Jersey

07901, United States

Yang Tang − Bristol Myers Squibb, San Diego, California

92121, United States

Proteomics. KG1 cells were treated with DMSO or 100 nM CC-

0009 for 4 h. All treatments were performed with four replicates and

protein lysates were digested with trypsin, labeled by isobaric tandem-

mass-tags (TMT), fractionated, and analyzed using an Orbitrap

Fusion Lumos mass spectrometer at IQ Proteomics (Cambridge,

MA). Peptide to protein mapping, protein identiﬁcation, and

quantiﬁcation of relative protein abundance with statistical analyses

were performed at Celgene using custom software. 7957 human

proteins were unambiguously quantiﬁed by at least one peptide in at

least one of the MS runs. Proteins were deﬁned by the Uniprot

identiﬁer, with some represented by more than one isoform.

Immunoblot Analysis. Cells were washed in ice-cold 1× PBS

twice before harvest in Buﬀer A [50 mM Tris·Cl (pH 7.6), 150 mM

NaCl, 1% Triton X-100, 1 mM EDTA, 1 mM EGTA, 1 mM β-

glycerophosphate, 2.5 mM sodium pyrophosphate, 1 mM Na3VO4, 1

μg/mL leupeptin, one tablet of Complete ULTRA protease inhibitor

cocktail (Roche), and one tablet of PhosSTOP phosphatase inhibitor

cocktail (Roche)]. Whole cell extracts were collected after

centrifugation at top speed for 10 min and total protein was

quantiﬁed using Bradford Reagent (Bio-Rad) according to the

manufacturer’s instructions. Then, 10−20 μg of total protein per

sample were resolved by SDS-PAGE gel electrophoresis, transferred

onto a nitrocellulose membrane using the Turboblot system (Bio-

Rad), and probed with the indicated primary antibodies. Bound

antibodies were detected with IRDye-680 or -800 conjugated

secondary antibodies using a LI-COR scanner.

Massimo Ammirante − Bristol Myers Squibb, San Diego,

California 92121, United States

Rama K. Narla − Bristol Myers Squibb, San Diego, California

92121, United States

Joseph R. Piccotti − Bristol Myers Squibb, San Diego,

California 92121, United States

Michael Pourdehnad − Bristol Myers Squibb, San Francisco,

California 94158, United States

Antonia Lopez-Girona − Bristol Myers Squibb, San Diego,

California 92121, United States

Complete contact information is available at:

https://pubs.acs.org/10.1021/acs.jmedchem.0c01489

Notes

The authors declare no competing ﬁnancial interest.

ACKNOWLEDGMENTS

■

We acknowledge Thomas Daniel, Greg Reyes, Neil Raheja,

and Katerina Leftheris for their encouraging support of this

work. We also acknowledge our compound management and

analytical groups for technical assistance and sample processing

as well as thank Dr. Ravi Kumar and his team at GVK Bio for

synthetic support.

ANIMAL USE

■

All animal studies were performed under protocols approved

by the Celgene Institutional Animal Care and Use Committee

842

J. Med. Chem. 2021, 64, 1835−1843

Journal of Medicinal Chemistry

Discovery and Structure-Activity Relationships of Novel Glutarimide

Drug Annotation

Analogs that Promote Degradation of Aiolos and/or GSPT1. J. Med.

Chem. 2018, 61 (2), 492−503.

ABBREVIATIONS

■

CELMoD, Cereblon E3 Ligase Modulating Drug; CRBN,

Cereblon; CUL4, cullin-4 protein; DDB1, damaged DNA-

binding protein 1; RBX1, RING box-domain protein; GSPT1,

G1 to S phase transition 1; DUB, deubiquitinating enzyme;

eRF3a, eukaryotic release factor 3a; NBS, N-bromosuccini-

mide; ACN, acetonitrile; HCl, hydrochloric acid; DMA,

dimethylacetamide; DMF, dimethylformamide; TLC, thin

layer chromatography; HPLC, high performance liquid

chromatography

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Elucidating the Mechanism of Action of CC-90009, a Novel Cereblon

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