AZD-5991 Racemate is the racemate of AZD-5991. AZD-5991 Racemate is a Mcl-1 inhibitor (antagonist) with IC50 of <3 nM as measured by FRET assay.

Physicochemical Properties

Molecular Formula	C35H34CLN5O3S2
Molecular Weight	672.2592
Exact Mass	671.179
CAS #	2143010-83-5
Related CAS #	AZD-5991;2143061-81-6;AZD-5991 (S-enantiomer);2143061-82-7
PubChem CID	131634760
Appearance	Typically exists as solid at room temperature
Density	1.42±0.1 g/cm3(Predicted)
Boiling Point	949.6±65.0 °C(Predicted)
LogP	6.8
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	1
Heavy Atom Count	46
Complexity	1060
Defined Atom Stereocenter Count	0
SMILES	ClC1C=CC2C3CCCOC4=CC(=CC5=CC=CC=C45)SCC4=CC(CSCC5C(=C(C)N(C)N=5)C=1C=2N(C)C=3C(=O)O)=NN4C
InChi Key	KBQCEQAXHPIRTF-UHFFFAOYSA-N
InChi Code	InChI=1S/C35H34ClN5O3S2/c1-20-31-29(38-40(20)3)19-45-17-22-15-23(41(4)37-22)18-46-24-14-21-8-5-6-9-25(21)30(16-24)44-13-7-10-26-27-11-12-28(36)32(31)33(27)39(2)34(26)35(42)43/h5-6,8-9,11-12,14-16H,7,10,13,17-19H2,1-4H3,(H,42,43)
Chemical Name	17-chloro-5,13,14,22-tetramethyl-28-oxa-2,9-dithia-5,6,12,13,22-pentazaheptacyclo[27.7.1.14,7.011,15.016,21.020,24.030,35]octatriaconta-1(36),4(38),6,11,14,16,18,20,23,29(37),30,32,34-tridecaene-23-carboxylic acid
Synonyms	AZD-5991 racemate; AZD5991; AZD 5991 racemate
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	Mcl-1 (Ki = 0.13 nM); Mcl-1 (IC50 = 0.7 nM)
ln Vitro	AZD5991 is a potent and direct inhibitor of Mcl-1 with high selectivity versus other Bcl-2 family proteins. Inducing rapid apoptosis in cancer cells, most notably myeloma and acute myeloid leukemia (GI50 < 100 nM), AZD5991 binds directly to Mcl-1 and activates the Bak-dependent mitochondrial apoptotic pathway. AZD5991 kills hematological cells more frequently than solid tumor cells in a panel of cancer-derived cell lines[1][3].
ln Vivo	After a single well-tolerated dose as monotherapy or in combination with bortezomib or venetoclax, AZD5991 exhibits significant antitumor activity in vivo with complete tumor regression in a number of models of multiple myeloma and acute myeloid leukemia. Cleavage of caspase-3 and PARP in these in vivo studies showed a strong correlation between AZD5991's cytotoxic activity and the activation of the mitochondrial apoptotic pathway[1].
Cell Assay	MOLP-8 cells are treated with AZD5991 or DMSO ontrol for 30 min. AZD5991 or DMSO ontrol are applied to MOLP-8 cells. The samples are then centrifuged, and the pellet is resuspended in ice-cold lysis buffer and incubated for 20 minutes on ice while vortexing every five minutes. After samples have been centrifuged, the protein concentration has been determined. Prior to overnight incubation with anti-Mcl-1 antibody at 4 °C with rotation, samples are pre-cleared for 30 min using rotation and a 50% slurry of Protein A/G magnetic beads. The next step is to add Protein A/G magnetic beads, which are rotated for 1 hour at 4 °C. Each IP pellet is then given a 10% sample reducing agent addition and four PBS:lysis buffer washes before being subjected to a western blotting analysis.
Animal Protocol	Mice and Rats[1] In mice, drugs (e.g., AZD5991; 10-100 mg/kg) are dosed intravenously in a volume of 5 mL/kg except for Venetoclax that is dosed orally in a volume of 10 mL/kg. One million MV4-11, five million MOLP-8, ten million NCI-H929 or five million OCI-AML3 cells are injected subcutaneously in the right flank of mice in a volume of 0.1 mL. In rats, AZD5991 (10-100 mg/kg) is dosed intravenously in a volume of 10 mL/kg. Ten million MV4-11 cells are injected subcutaneously in the right flank of rats in a volume of 0.1 mL. Tumor volumes (measured by caliper), animal body weight, and tumor condition are recorded twice weekly for the duration of the study. The tumor volume is calculated[1].
References	[1]. Nat Commun . 2018 Dec 17;9(1):5341. [2]. Curr Opin Chem Biol . 2017 Aug:39:133-142. [3]. Cancer Res (2018) 78 (13_Supplement): 302.
Additional Infomation	AZD-5991 is under investigation in clinical trial NCT03218683 (Study of AZD5991 in Relapsed or Refractory Haematologic Malignancies.). Mcl-1 Inhibitor AZD5991 is an inhibitor of induced myeloid leukemia cell differentiation protein (myeloid cell leukemia-1; Mcl-1; Bcl2-L-3), with potential pro-apoptotic and antineoplastic activities. Upon administration, AZD5991 binds to Mcl-1, thereby preventing the binding of Mcl-1 to and inactivation of certain pro-apoptotic proteins, and promoting apoptosis of cells overexpressing Mcl-1. Mcl-1, an anti-apoptotic protein belonging to the Bcl-2 family of proteins, is upregulated in cancer cells and promotes tumor cell survival.

Solubility Data

Solubility (In Vitro)

DMSO: ~100 mg/mL (~148.8 mM)

Solubility (In Vivo)

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. Injection Formulations (e.g. IP/IV/IM/SC) 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). 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) Oral Formulations 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). 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 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.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.4875 mL	7.4376 mL	14.8752 mL
	5 mM	0.2975 mL	1.4875 mL	2.9750 mL
	10 mM	0.1488 mL	0.7438 mL	1.4875 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.