RMC-5552 is a potent and specific mTORC1 inhibitor. RMC-5552 inhibits the phosphorylation of mTORC1 pS6K and p4EBP1 with IC50 of 0.14 nM and 0.48 nM, respectively. RMC-5552 displays much lower inhibition of pAKT (IC50 of 19 nM), resulting in mTORC1/mTORC2 selectivity approaching 40-fold. RMC-5552 has anti-cancer effect.

Physicochemical Properties

Molecular Formula	C93H136N10O24
Molecular Weight	1778.13
Exact Mass	1776.972
CAS #	2382768-62-7
PubChem CID	162640850
Appearance	White to off-white solid powder
LogP	7
Hydrogen Bond Donor Count	6
Hydrogen Bond Acceptor Count	30
Rotatable Bond Count	38
Heavy Atom Count	127
Complexity	3460
Defined Atom Stereocenter Count	16
SMILES	C1C=C(C)[C@H](C[C@@]2(CC[C@H]([C@@](O)(O2)C(C(N2CCCC[C@@]2([H])C(=O)O[C@]([H])([C@@H](C[C@@H]2CC[C@H]([C@H](OC)C2)OC(=O)NCCOCCOCCOCCOCCOCCOCCOCCOCCC(=O)N2CC3=CC=C(CN4N=C(C5C6N=C(N)OC=6C=CC=5)C5C(N)=NC=NC4=5)C=C3CC2)C)C[C@@H](O)[C@@H](C=C([C@@H](O)[C@@H](OC)C(=O)[C@H](C)C[C@H](C)C=CC=1)C)C)=O)=O)C)[H])OC |c:1,t:114,128,130|
InChi Key	IUTYTXDIGUNPAA-GKVGALOGSA-N
InChi Code	InChI=1S/C93H136N10O24/c1-59-16-12-11-13-17-60(2)77(113-8)54-71-24-19-65(7)93(112,127-71)86(108)89(109)102-30-15-14-18-73(102)90(110)124-78(55-74(104)61(3)49-64(6)84(107)85(115-10)83(106)63(5)48-59)62(4)50-66-21-25-76(79(52-66)114-9)126-92(111)96-29-33-117-35-37-119-39-41-121-43-45-123-47-46-122-44-42-120-40-38-118-36-34-116-32-28-80(105)101-31-27-68-51-67(20-22-70(68)57-101)56-103-88-81(87(94)97-58-98-88)82(100-103)69-23-26-75-72(53-69)99-91(95)125-75/h11-13,16-17,20,22-23,26,49,51,53,58-59,61-63,65-66,71,73-74,76-79,84-85,104,107,112H,14-15,18-19,21,24-25,27-48,50,52,54-57H2,1-10H3,(H2,95,99)(H,96,111)(H2,94,97,98)/b13-11+,16-12+,60-17+,64-49+/t59-,61-,62-,63-,65-,66+,71+,73+,74-,76-,77+,78+,79-,84-,85+,93-/m1/s1
Chemical Name	[(1R,2R,4S)-2-methoxy-4-[(2R)-2-[(1R,9S,12S,14R,15R,16E,18R,19R,21R,23S,24E,26E,28E,30S,32S,35R)-1,14,18-trihydroxy-19,30-dimethoxy-15,17,21,23,29,35-hexamethyl-2,3,10,20-tetraoxo-11,36-dioxa-4-azatricyclo[30.3.1.04,9]hexatriaconta-16,24,26,28-tetraen-12-yl]propyl]cyclohexyl] N-[2-[2-[2-[2-[2-[2-[2-[2-[3-[6-[[4-amino-3-(2-amino-1,3-benzoxazol-5-yl)pyrazolo[3,4-d]pyrimidin-1-yl]methyl]-3,4-dihydro-1H-isoquinolin-2-yl]-3-oxopropoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethyl]carbamate
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

ln Vitro	Only in the presence of the FKBP12-FRB allosteric mode of RMC-5552 was recruitment of FKBP12 seen. At the interface between FKBP12 and the FRB domain of mTOR, the density of RMC-5552 is seen. Through the 4-aminopyrazolo[3,4-d] braid core, the “Manhattan chain” of mTOR, and the 2-aminobenzoxazole to E2190 and K2187 Reply[1], RMC-5552 forms hydrogen connections with the backbone of G2238 and V2240.
ln Vivo	RMC-5552 (1-10 mg/kg; intraperitoneal injection; once weekly; for 28 days) displayed anti-activity in a mouse MCF-7 human breast cancer xenograft model [1]. RMC-5552 demonstrates anti-activity in mice [1]. PK Parameters in Perfume 1 mg/kg[1] Compound Tmax (h) Cmax (ng/mL) Cmax (μM) AUClast (μg/mL × h) AUClast (μM × h) t1/2 (h) ) 38 RMC-5552 2.0 ± 0.0 5667 ± 1106 3.19 ± 0.62 46 089 ± 5320 25.9 ± 3.0 4.8 ± 0.4
Animal Protocol	Animal/Disease Models: Female Balb/c nude mice (6-8 weeks old) injected with MCF-7 cells [1] Doses: 1 mg/kg, 3 mg/kg, 10 mg/kg. Route of Administration: intraperitoneal (ip) injection; once a week; for 28 days Experimental Results: resulted in tumor volume reduction.
References	[1]. Discovery of RMC-5552, a Selective Bi-Steric Inhibitor of mTORC1, for the Treatment of mTORC1-Activated Tumors. J Med Chem. 2022 Dec 19.
Additional Infomation	mTORC1 Kinase Inhibitor RMC-5552 is a bi-steric inhibitor of the raptor-mammalian target of rapamycin (mTOR) complex 1 (mTOR complex 1; mTORC1), with potential antineoplastic activity. Upon administration, mTORC1 kinase inhibitor RMC-5552 selectively targets, binds to and inhibits the serine/threonine kinase activity of mTORC1, resulting in decreased expression of mRNAs necessary for cell cycle progression, which may induce cell cycle arrest and tumor cell apoptosis. mTORC1 phosphorylates transcription factors, such as ribosomal protein S6 kinase beta-1 (S6K1) and eukaryotic translation initiation factor 4E-binding protein 1 (4EBP1), which stimulate protein synthesis and regulate cell growth, proliferation, motility, and survival. RMC-5552 is able to suppress the phosphorylation of the tumor suppressor activity of 4EBP1, induces apoptosis and decreases proliferation of mTORC1-expressing tumor cells. mTORC1 is a serine/threonine kinase that is upregulated in certain tumor cell types. It plays an important role in the PI3K/Akt/mTOR signaling pathway, which is often deregulated in cancer cells and promotes cell growth, survival, and resistance to chemotherapy and radiotherapy. Selective mTORC1 inhibition improves the poor clinical tolerability as compared to agents that also cause inhibition of the mTORC2 complex.

Solubility Data

Solubility (In Vitro)

DMSO : ~166.67 mg/mL (~93.73 mM)

Solubility (In Vivo)

Solubility in Formulation 1: 5 mg/mL (2.81 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 50.0 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: 5 mg/mL (2.81 mM) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 50.0 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	0.5624 mL	2.8119 mL	5.6239 mL
	5 mM	0.1125 mL	0.5624 mL	1.1248 mL
	10 mM	0.0562 mL	0.2812 mL	0.5624 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.