Sirpiglenastat (DRP-104) is a broadly acting glutamine antagonist. Sirpiglenastat has anti-neoplastic activities by directly targeting tumor metabolism and simultaneously inducing a potent anti-tumor immune response.

Physicochemical Properties

Molecular Formula	C22H27N5O5
Molecular Weight	441.480284929276
Exact Mass	441.201
CAS #	2079939-05-0
PubChem CID	137308771
Appearance	Off-white to yellow solid powder
LogP	1.2
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	6
Rotatable Bond Count	12
Heavy Atom Count	32
Complexity	749
Defined Atom Stereocenter Count	2
SMILES	C(C1=CNC2=CC=CC=C12)[C@H](NC(=O)C)C(=O)N[C@@H](CCC(=O)C=[N+]=[N-])C(=O)OC(C)C
InChi Key	LQNMCWOJACNQQM-PMACEKPBSA-N
InChi Code	InChI=1S/C22H27N5O5/c1-13(2)32-22(31)19(9-8-16(29)12-25-23)27-21(30)20(26-14(3)28)10-15-11-24-18-7-5-4-6-17(15)18/h4-7,11-13,19-20,24H,8-10H2,1-3H3,(H,26,28)(H,27,30)/t19-,20-/m0/s1
Chemical Name	propan-2-yl (2S)-2-[[(2S)-2-acetamido-3-(1H-indol-3-yl)propanoyl]amino]-6-diazo-5-oxohexanoate
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 Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light.
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

ln Vitro	Treatment with siropilenastat (DRP-104) exhibits extensive effects on immune cell modulation, such as enhanced T, NK, and macrophages. Pro-tumorigenic cytokines including VEGF and KC (IL-8) are reduced by sirpiglenastat[1].
ln Vivo	At day 12, 90% tumor growth inhibition is seen in CT26 bearing mice treated with Sirpiglenastat (DRP-104) (0.5 mg/kg; sc; once daily; for five days). 36 days are the median survival time[1]. In the H22 model, therapy with sirpiglenastat (0.5 mg/kg; sc) effectively suppresses tumor growth[1].
Animal Protocol	Animal/Disease Models: CT26 bearing mice[1] Doses: 0.5 mg/kg Route of Administration: sc; one time/day; for 5 days Experimental Results: demonstrated tumor growth inhibition in mice.
References	[1]. DRP-104, a novel broad acting glutamine antagonist, induces distinctive immune modulation mechanisms and synergistic efficacy in combination with immune checkpoint blockade. J Immunother Cancer. 2019 Nov 6;7(Suppl 1):282. [2]. DRP-104, a broad acting glutamine antagonist, synergizes with immune checkpoint blockade in vivo. Cancer Res 2021;81(13_Suppl):Abstract nr 1563.
Additional Infomation	Sirpiglenastat, also known as DRP-104, is a prodrug of [6-diazo-5-oxo-L-norleucine] (DON), which is a broad-acting glutamine antagonist. It is under investigation in clinical trial NCT04471415 (Study to Investigate DRP-104 in Adults With Advanced Solid Tumors). Sirpiglenastat is a broad acting glutamine antagonist, with potential immunomodulatory and antineoplastic activities. Upon administration, DON (6-Diazo-5-oxo-L-norleucine), the active moiety of sirpiglenastat, irreversibly inhibits multiple enzymes involved in glutamine metabolism. Blocking glutamine metabolism inhibits proliferation in rapidly growing tumor cells and leads to an induction of cell death. Unlike normal healthy cells, glutamine-dependent tumors rely heavily on the intracellular conversion of exogenous glutamine into glutamate and glutamate metabolites to both provide energy and generate building blocks for the production of macromolecules, which are needed for cellular growth and survival. In addition, blocking glutamine metabolism leads to the accumulation of glutamine in tumor cells and increases glutamine concentration in the tumor microenvironment (TME) upon tumor cell death. As glutamine is essential for T-cell generation, DON may also enhance T-cell proliferation and activation in the TME, which may lead to further killing of tumor cells. The conversion of sirpiglenastat to the active moiety DON occurs primarily in tumor cells, allowing glutamine metabolism in healthy cells which may lessen adverse effects.

Solubility Data

Solubility (In Vitro)

DMSO: 100 mg/mL (226.51 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	2.2651 mL	11.3255 mL	22.6511 mL
	5 mM	0.4530 mL	2.2651 mL	4.5302 mL
	10 mM	0.2265 mL	1.1326 mL	2.2651 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.