E-Selectin (Kd = 0.46 uM; IC50 = 1.75 uM)

Uproleselan (GMI-1271) is a novel E-selectin antagonist that disrupts cell survival pathways, enhances chemotherapy response, improves survival in mouse xenograft and syngeneic models, and decreases chemotherapy toxicity in vivo. A phase 1/2 study evaluated the safety, tolerability, and antileukemic activity of uproleselan (5-20 mg/kg) with MEC (mitoxantrone, etoposide, and cytarabine) among patients with relapsed/refractory (R/R) acute myeloid leukemia (AML). Among the first 19 patients, no dose-limiting toxicities were observed. The recommended phase 2 dose (RP2D) was 10 mg/kg twice daily. An additional 47 patients with R/R AML were treated with uproleselan at the RP2D plus MEC. At the RP2D, the remission rate (complete response [CR]/CR with incomplete count recovery [CRi]) was 41% (CR, 35%), and the median overall survival (OS) was 8.8 months. In a separate cohort, 25 newly diagnosed patients age ≥60 years received uproleselan at the RP2D plus cytarabine and idarubicin (7 + 3). In these frontline patients, the CR/CRi rate was 72% (CR, 52%), and the median OS was 12.6 months. The addition of uproleselan was associated with low rates of oral mucositis. E-selectin ligand expression on leukemic blasts was higher in patients with relapsed vs primary refractory AML and in newly diagnosed older patients with high-risk cytogenetics and secondary AML. In the R/R cohort, E-selectin expression >10% was associated with a higher response rate and improved survival. The addition of uproleselan to chemotherapy was well tolerated, with high remission rates, low induction mortality, and low rates of mucositis, providing a strong rationale for phase 3 randomized confirmatory studies. This trial was registered at www.clinicaltrials.gov as #NCT02306291.[1]

In all study phases, uproleselan was administered as a 20-minute IV infusion given 24 hours prior, twice daily throughout, and twice daily for 48 hours postinduction chemotherapy. The salvage chemotherapy regimen used for patients with R/R disease was MEC (10 mg/m2 of mitoxantrone per day IV over 15-20 minutes, 100 mg/m2 of etoposide per day IV over 60 minutes, and 1000 mg/m2 of cytarabine per day IV over 60 minutes for 5 days) for 1 induction cycle. Uproleselan dose levels were determined by targeting a range of expected exposures, at or above the pharmacologically active dose range, as demonstrated in preclinical models. Based on available PK data from nonhuman primate studies and safety data from human volunteer studies, the starting dose for uproleselan administered with MEC in the dose-escalation phase of the study was 5 mg/kg. Uproleselan was administered across 3 pharmacologically active dose levels (5, 10, and 20 mg/kg). Dose escalation was performed in the absence of dose-limiting toxicity (DLT), defined as myelosuppression (failure of recovery to absolute neutrophil count [ANC] ≥0.5 × 109/L and platelet count of ≥25 × 109/L) beyond day 42 in the absence of persistent morphologic evidence of leukemia in the marrow or grade 3 nonhematologic toxicity attributable to uproleselan and not resolving to grade 2 by day 42. The dose-escalation committee determined an RP2D of 10 mg/kg, defined as the dose that (1) did not cause DLT in >33% of treated patients during induction and (2) was the most appropriate dose for continuing clinical evaluation in AML based on available data (safety, exposure achieved, pharmacodynamic [PD] assessment of on-target effect, clinical activity, and overall toxicity).[1]

The phase 2 dose expansion at the RP2D continued enrolling patients with R/R AML. Responding patients were eligible to receive 1 additional cycle of consolidation with uproleselan combined with a 4-day course of MEC. Alternatively, patients underwent hematopoietic stem cell transplantation and/or received additional postremission therapies at the discretion of their treating physician.[1]

A second cohort of newly diagnosed patients age ≥60 years were enrolled between June 2016 and February 2017 and treated with uproleselan administered at the RP2D in combination with conventional 7 + 3 induction chemotherapy (200 mg/m2 of cytarabine by a 24-hour continuous daily infusion on days 1-7 in combination with 12 mg/m2 of idarubicin by IV bolus daily on days 1-3). Uproleselan was administered on the same schedule as that used in the R/R AML cohort. For patients with residual leukemia detected on a day-15 midcycle bone marrow examination, a second cycle of induction therapy (5 + 2) was allowed in combination with uproleselan at the same dose and schedule as those used during the initial 7 + 3 induction. The first 3 patients were assessed postinduction for DLT; thereafter, enrollment was opened to complete a 25-patient cohort. Responders could receive consolidation therapy with uproleselan plus intermediate-dose cytarabine (2000 mg/m2 per day over 3 hours for 5 days or 1500 mg/m2 over 3 hours every 12 hours on days 1, 3, and 5) for up to 3 cycles. Patients could undergo hematopoietic stem cell transplantation at the discretion of their treating physician.[1]

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Safety assessments[1]
The primary end point of the study was the frequency, severity, and relatedness of treatment-emergent adverse events (TEAEs) in patients receiving uproleselan in combination with chemotherapy. Safety assessments included the surveillance and recording of TEAEs, vital sign measurements, clinical laboratory tests, ECOG performance status, and physical examinations. Grade and term of TEAEs were reported by the treating physician and were graded by the Common Terminology Criteria for Adverse Events (version 4.03).

Dose-escalation cohort[1]
Nineteen patients in the dose-escalation phase were enrolled in 3 separate uproleselan dose-level groups of 5 (n = 6), 10 (n = 7), and 20 mg/kg (n = 6). Uproleselan at 10 mg/kg twice daily was established as the RP2D and was used as the dose for combination therapy with MEC in the dose-expansion phase and with idarubicin and cytarabine in the newly diagnosed cohort. No DLTs were observed at any dose level in phase 1; therefore, DLT did not influence dose-level selection for phase 2. PD analysis demonstrated on-target activity as measured by reduction in shed soluble E-selectin in the plasma at all 3 dose levels. Furthermore, no dose response was observed, suggesting that all 3 dose levels assessed may have exceeded a plateau level for PD effect. Clinical outcomes (bone marrow response to uproleselan with MEC-induction chemotherapy) were similar across the dose levels. Based on the PK analysis in the phase 1 portion of this study, 10 mg/kg of uproleselan provided the highest levels of exposure that did not exceed the 14-day nonclinical safety limits, and this dose was selected for further testing in the phase 2 portion of the study.

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R/R AML cohort[1]
For the entire cohort of patients with R/R AML, the incidence of TEAEs, regardless of relationship to study drug, was similar across all uproleselan dose levels (supplemental Table 1). None of the 66 patients with R/R AML discontinued treatment because of an AE. As expected in patients with AML, all patients in this R/R cohort had evidence of grade 4 myelosuppression (thrombocytopenia, neutropenia, or anemia) during the study. For patients achieving a response (complete response [CR] or CR with incomplete count recovery [CRi]), the median time to count recovery (ANC ≥500/μL and platelets ≥50/μL) was 33.0 days (90% CI, 31.0-34.0). Table 2 shows the overall incidence of grade 3 or 4 TEAEs. Most of the TEAEs observed were typical of the background chemotherapy, with few TEAEs attributed to uproleselan (supplemental Table 2 provides an overview of TEAEs by dose). Apart from hematologic toxicities, the only grade 3 or 4 TEAEs reported for ≥10% of patients was sepsis (12% of patients). Gastrointestinal toxicities, including nausea, vomiting, diarrhea, and colitis, were mild, with grade 3 events occurring in <5% of patients. Hepatic and renal TEAEs were mostly grade 1 or 2, with grade 3 or 4 observed in 5% and 5% of patients, respectively. One patient (2%) died within 30 days of initiation of therapy; the 60-day mortality rate was 9% (n = 6). Grade 3 mucositis was reported in only 2% of patients. Other nonhematologic TEAEs were mild and generally judged to be unrelated to uproleselan.

 Blood. 2021 Sep 20;blood.2021010721. doi: 10.1182/blood.2021010721.

Uproleselan is a novel, specific E-Selectin antagonist under investigation in clinical trial NCT02306291 (Study to Determine Safety, Pharmacokinetics and Efficacy of GMI-1271 in Combination With Chemotherapy in AML).
Uproleselan is a synthetic, glycomimetic molecule and E-selectin (CD62E) antagonist, with potential anti-thrombotic, antineoplastic and chemopotentiating activities. Upon administration, uproleselan binds to E-selectin expressed on endothelial cells and prevents their interaction with selectin-E ligand-expressing cancer cells. This may prevent tumor cell activation, migration and metastasis. GMI-1271 also interferes with the binding of selectin E-expressing vascular endothelial cells to selectin-E ligand-expressing monocytes and neutrophils, thereby disrupting their activation. Consequently, this inhibits both the activation of the coagulation cascade and thrombus formation. This agent also prevents both leukocyte activation and inflammation. E-selectin is a cell adhesion molecule involved in cell rolling, signaling and chemotaxis; it also plays a crucial role in inflammatory processes and cancer.

C60H109N3O28

1320.52

1319.719

1983970-12-2

1983970-12-2 (free acid); 1914993-95-5 (sodium)

71600085

Typically exists as solid at room temperature

1.27±0.1 g/cm3(Predicted)

1249.6±65.0 °C(Predicted)

-2

9

27

52

90

1870

15

O([C@H]1[C@@H]([C@H]([C@H]([C@@H](CO)O1)O)O[C@H](C(=O)O)CC1CCCCC1)NC(C)=O)[C@@H]1C[C@H](C(NCCNC(CCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOC)=O)=O)C[C@H](CC)[C@H]1O[C@H]1[C@H]([C@@H]([C@@H]([C@H](C)O1)O)O)O

ALGJTRWKCMYWHE-YFEAQVBDSA-N

InChI=1S/C60H109N3O28/c1-5-44-38-45(39-46(53(44)90-58-52(69)51(68)50(67)41(2)87-58)88-59-54(63-42(3)65)60(74,55(70)48(40-64)89-59)91-47(57(72)73)37-43-9-7-6-8-10-43)56(71)62-13-12-61-49(66)11-14-76-17-18-78-21-22-80-25-26-82-29-30-84-33-34-86-36-35-85-32-31-83-28-27-81-24-23-79-20-19-77-16-15-75-4/h41,43-48,50-55,58-59,64,67-70,74H,5-40H2,1-4H3,(H,61,66)(H,62,71)(H,63,65)(H,72,73)/t41-,44-,45+,46+,47+,48+,50+,51+,52-,53+,54-,55-,58?,59?,60-/m0/s1

(2R)-2-(((3S,4S,5S,6R)-3-acetamido-2-(((1R,2R,3S,5R)-3-ethyl-5-((38-oxo-2,5,8,11,14,17,20,23,26,29,32,35-dodecaoxa-39-azahentetracontan-41-yl)carbamoyl)-2-(((3S,4R,5S,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)cyclohexyl)oxy)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-4-yl)oxy)-3-cyclohexylpropanoic acid

GMI-1271; Uproleselan; GMI-1271; 1983970-12-2; Uproleselan [USAN]; GMI-1271 FREE ACID; PE952ANF83; UPROLESELAN [INN]; Uproleselan (USAN/INN); GMI1271 Uproleselan

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

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.

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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).

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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)

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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).

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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

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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.

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 (Please use freshly prepared in vivo formulations for optimal results.)