E-Selectin (Kd = 0.46 uM; IC50 = 1.75 uM)
The primary target of Uproleselan is E-selectin (also known as CD62E), a cell adhesion molecule expressed on vascular endothelial cells. E-selectin is constitutively expressed in the bone marrow microenvironment and is also inducibly expressed during inflammatory responses. In acute myeloid leukemia (AML), alterations in cell-surface glycosylation associated with oncogenesis lead to overexpression of E-selectin ligands (sialyl Lewisˣ) on AML blasts, enhancing binding to E-selectin. This binding directly promotes chemoresistance and survival of AML cells through activation of pro-survival signaling pathways such as AKT/NF-κB. As a carbohydrate analog of sialyl Lewisˣ, Uproleselan binds to E-selectin with high affinity (IC50 of 2.4 µM), competitively blocking the interaction between E-selectin and leukemia cells, thereby attenuating the protective effect of the bone marrow microenvironment on AML cells. In terms of selectivity, Uproleselan demonstrates excellent selectivity for E-selectin over P-selectin (IC50 >10,000 µM) and L-selectin (IC50 4,516 µM).
Uproleselan targets E-selectin (also known as CD62E), a cell adhesion molecule expressed on vascular endothelial cells. E-selectin is constitutively expressed in the bone marrow microenvironment. In acute myeloid leukemia (AML), oncogenesis leads to overexpression of E-selectin ligands (sialyl Lewisˣ) on AML blasts. The binding of these ligands to E-selectin directly promotes chemoresistance and survival of AML cells through pro-survival signaling pathways such as AKT/NF-κB. Uproleselan binds to E-selectin with high affinity, competitively blocking this interaction.

Uproleselan demonstrates high-affinity E-selectin binding activity and functional blocking activity in vitro. Surface plasmon resonance (SPR) assays show that Uproleselan binds to human E-selectin with a KD of approximately 61 nM and an IC50 of approximately 800 nM, effectively blocking the binding of sialyl Lewisˣ to E-selectin. In functional assays, Uproleselan reverses bone marrow microenvironment-induced chemoresistance in AML cells by disrupting E-selectin-mediated pro-survival signaling pathways. In vitro studies demonstrate that adhesion to E-selectin activates AKT and NF-κB signaling pathways in AML cells, promoting cell survival; the addition of Uproleselan effectively inhibits this signal transduction and increases the sensitivity of leukemic cells to chemotherapeutic agents. Furthermore, Uproleselan induces mobilization of AML cells from the protective bone marrow niche, though this effect accounts for only approximately 1% of total cell numbers, suggesting its primary effect is chemosensitization rather than pure leukemia cell mobilization.

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In vitro studies show that Uproleselan binds to E-selectin with high affinity, with a Kd of 0.46 µM and an IC50 of 1.75 µM. It competitively blocks the interaction between E-selectin and leukemia cells, thereby attenuating the protective effect of the bone marrow microenvironment on AML cells. As a carbohydrate analog of sialyl Lewisˣ, it demonstrates excellent selectivity for E-selectin.

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]

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Uproleselan demonstrates significant anti-leukemia activity in various in vivo models. In an AML mouse model, Uproleselan combined with chemotherapy (cytarabine) doubled mouse survival duration and reduced the number of chemotherapy-surviving leukemia regenerating cells (LRC) by 4-10 fold. In an MV4-11 AML xenograft mouse model, Uproleselan combined with chemotherapy achieved significant leukemia cell clearance. In a venous thrombosis model, Uproleselan (40 mg/kg, intraperitoneal injection) significantly reduced thrombus weight (median 0.0138 g vs vehicle control 0.0208 g). In a non-human primate iliac vein thrombosis model, Uproleselan (25 mg/kg, subcutaneous, once daily) for 21 days showed the highest vein recanalization rate without causing clinically significant coagulation changes or bleeding events. In clinical trials, Uproleselan combined with MEC chemotherapy (mitoxantrone, etoposide, and cytarabine) in relapsed/refractory AML patients achieved an overall response rate of 41% (CR 35%) and median overall survival of 8.8 months; in newly diagnosed older AML patients receiving “7+3” induction, the overall response rate was 72% (CR 52%) with median overall survival of 12.6 months.

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In vivo, Uproleselan has demonstrated the ability to disrupt cell survival pathways and enhance chemotherapy response. It improves survival in mouse xenograft and syngeneic models and decreases chemotherapy-related toxicity. By blocking the adhesive interaction between leukemia cells and the bone marrow vascular endothelium, Uproleselan overcomes chemotherapy resistance and enhances anti-leukemia efficacy. It also prevents leukocyte activation and inflammation, contributing to its anti-thrombotic activity.

The E-selectin binding affinity of Uproleselan is assessed using Surface Plasmon Resonance (SPR) and Microscale Thermophoresis (MST) techniques. A typical SPR assay protocol is as follows: Recombinant human E-selectin protein is immobilized on a CM5 chip surface. Increasing concentrations of Uproleselan (0.1 nM–100 μM) are prepared in HBS-EP+ buffer (10 mM HEPES pH 7.4, 150 mM NaCl, 3 mM EDTA, 0.05% Tween-20) and flowed over the chip surface at 30 μL/min, with a 2-3 minute association phase and a 5-10 minute dissociation phase. Surface plasmon resonance signal changes are monitored using a Biacore or ProteOn instrument, and data are fitted using a 1:1 Langmuir binding model to calculate association rate constant (ka), dissociation rate constant (kd), and equilibrium dissociation constant (KD). Functional blocking activity is assessed by competition ELISA: E-selectin-Fc fusion protein is coated onto 96-well plates, biotin-labeled sialyl Lewisˣ and increasing concentrations of Uproleselan (0.1 nM–100 μM) are added, and bound ligand is detected using streptavidin-HRP to calculate IC50 values. Uproleselan exhibits an IC50 of approximately 800 nM for E-selectin. Additionally, radioligand binding assays or cell-based calcium flux assays can be used to further validate selectivity.

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Specific cell-free binding assay protocols for Uproleselan likely involve surface plasmon resonance (SPR) or enzyme-linked immunosorbent assays (ELISA) to measure its binding affinity to recombinant E-selectin protein. These assays determine key parameters such as the dissociation constant (Kd) and the half-maximal inhibitory concentration (IC50) for the compound's interaction with its target.

In vitro cellular activity of Uproleselan is primarily assessed by evaluating its blocking effect on leukemia cell adhesion to E-selectin and its enhancement of chemosensitivity. A typical assay protocol is as follows: Human umbilical vein endothelial cells (HUVECs) or CHO cells stably expressing human E-selectin are cultured in medium containing 10% fetal bovine serum in a 5% CO₂ incubator at 37°C. Endothelial cells are seeded in 96-well plates (2×10⁴ cells per well) and cultured to confluence, then stimulated with 1 U/mL thrombin or TNF-α (10 ng/mL) for 4-6 hours to induce E-selectin expression. Fluorescently labeled AML cell lines (such as KG-1a, OCI-AML3, 1×10⁵ cells per well) are added along with increasing concentrations of Uproleselan (0.1 nM–10 μM), and incubated at 37°C for 30-60 minutes. After gentle washing to remove non-adherent cells, adherent cells are quantified using a fluorescence plate reader (excitation/emission 485/535 nm) to calculate IC50 values. Chemosensitization assays are performed under conditions of E-selectin-Fc-coated plates or co-culture with E-selectin+ stromal cells: AML cells are treated with cytarabine (0.1-10 μM) and Uproleselan (5-20 μM) for 48-72 hours, and apoptosis rates are assessed by Annexin V/PI double staining flow cytometry or cell viability by MTS/CCK-8 assay. Studies have shown that Uproleselan effectively inhibits E-selectin-mediated leukemia cell adhesion and reverses stroma-mediated chemoresistance.

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In vitro cell-based assays for Uproleselan are conducted using AML cell lines that express E-selectin ligands. Cells are treated with the compound, and its ability to inhibit cell adhesion to E-selectin-coated surfaces is assessed. The compound's effect on downstream pro-survival signaling pathways, such as AKT/NF-κB, can also be evaluated by measuring the phosphorylation status of key proteins via Western blot.

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).

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The in vivo efficacy of Uproleselan is evaluated using various animal models. In AML mouse models, 8-12 week old female NSG or C57BL/6J mice are injected via the tail vein with MV4-11 or MLL-AF9 AML cells (1×10⁶-5×10⁶ per mouse). After leukemia establishment (approximately 1-3 weeks), mice are randomized into groups: vehicle control group (PBS), chemotherapy group (cytarabine 100 mg/kg, intraperitoneal injection, days 1-5), Uproleselan monotherapy group (25-50 mg/kg, intraperitoneal injection, once or twice daily), and combination therapy group. Leukemia burden is monitored by bioluminescence imaging (BLI), and survival curves are recorded to assess overall survival. In a venous thrombosis model, male C57BL/6J mice are anesthetized with isoflurane and undergo laparotomy to expose the inferior vena cava (IVC). Electrodes are placed around the IVC, and a 250 mA electrical current is applied to induce non-occlusive thrombosis. Immediately after thrombus induction, a single intraperitoneal injection of Uproleselan (40 mg/kg) or vehicle control is administered, and animals are euthanized 24 hours later for IVC harvest and thrombus weight measurement. In a non-human primate deep vein thrombosis model, thrombosis is induced by 6-hour balloon occlusion of the iliac vein. Starting on day 2 post-thrombosis, Uproleselan is administered at 25 mg/kg subcutaneously once daily for 21 consecutive days. Vein recanalization rate is assessed by magnetic resonance venography (MRV), vascular patency by Doppler ultrasound, and coagulation parameters (prothrombin time, activated partial thromboplastin time) are monitored.

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In vivo animal studies for Uproleselan have been performed using mouse xenograft and syngeneic models. In these studies, mice bearing tumors are treated with Uproleselan, often in combination with chemotherapy. Key endpoints include tumor growth inhibition, survival rates, and assessment of chemotherapy-related toxicity.

The pharmacokinetic profile of Uproleselan has been characterized through multiple preclinical and clinical studies. In healthy volunteers, intravenously administered Uproleselan exhibits favorable pharmacokinetic characteristics, with key parameters including: time to peak concentration (Tmax) of approximately 1-2 hours, dose-dependent peak concentration (Cmax), and an elimination half-life (T₁/₂) of several hours. The apparent volume of distribution (Vz) suggests the drug primarily distributes in blood and extracellular spaces. The primary elimination pathways are still under investigation, but renal clearance plays a role, with parent drug excretion detectable in urine. An ongoing Phase I study is evaluating the pharmacokinetics of Uproleselan in combination with chemotherapy in Chinese patients with relapsed/refractory AML, including parameters such as Cmax, Tmax, AUC0-12, and AUC0-t. Functional E-selectin blockade is achieved at a plasma concentration of 38 µM in patients. No significant pharmacokinetic drug-drug interactions have been reported for Uproleselan to date. The recommended Phase II dose (RP2D) of Uproleselan is 10 mg/kg administered intravenously twice daily.

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Detailed pharmacokinetic properties of Uproleselan are not extensively reported in the provided literature. The compound has a large molecular weight of 1304.53 g/mol and a molecular formula of C60H109N3O27. It is being investigated in clinical trials, indicating that it has favorable drug-like properties for systemic administration.

Dose escalation cohort [1]
In the dose escalation phase, a total of 19 patients were enrolled and divided into three different uproleselan dose groups: 5 mg/kg (n = 6), 10 mg/kg (n = 7), and 20 mg/kg (n = 6). Uproleselan at 10 mg/kg twice daily was identified as RP2D and was used in combination with MEC in the dose expansion phase, as well as in combination with idarubicin and cytarabine in the newly diagnosed cohort. No dose-limiting toxicities (DLTs) were observed in any of the dose groups in the phase 1 trial; therefore, DLTs did not affect the dose selection in the phase 2 trial. Pharmacodynamic (PD) analysis showed that the release of soluble E-selectin in plasma was reduced in all three dose groups, indicating that the drug had targeting activity. In addition, no dose response was observed, suggesting that the three dose groups evaluated may have exceeded the plateau of PD effects. Clinical outcomes (bone marrow response to proroleselan in combination with MEC-induced chemotherapy) were similar across the dose groups. Based on the pharmacokinetic analysis of the Phase I portion of this study, the 10 mg/kg procelandine dose group provided the highest level of drug exposure without exceeding the 14-day non-clinical safety limit, and therefore this dose was selected for further testing in the Phase II portion.

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Relapsed/Refractory Acute Myeloid Leukemia (R/R AML) Cohort [1]
For all R/R AML patients, regardless of whether it was related to the study drug, the incidence of adverse events (TEAEs) during treatment was similar across all procelandine dose groups (Supplementary Table 1). None of the 66 R/R AML patients discontinued treatment due to adverse events. As expected, all patients in this R/R cohort experienced Grade 4 myelosuppression (thrombocytopenia, neutropenia, or anemia) during the study period. For patients achieving remission (complete remission [CR] or complete remission with incomplete recovery of blood cell counts [CRi]), the median time to recovery of blood cell counts (absolute neutrophil count ≥500/μL and platelet count ≥50/μL) was 33.0 days (90% CI, 31.0–34.0). Table 2 shows the overall incidence of grade 3 or 4 treatment-interventional adverse events (TEAEs). Most observed TEAEs were typical adverse events of background chemotherapy, with only a few TEAEs attributed to uproleselan (Supplementary Table 2 provides an overview of TEAEs by dose). Apart from hematologic toxicities, the only grade 3 or 4 TEAE with an incidence ≥10% was sepsis (12% of patients). Gastrointestinal toxicities, including nausea, vomiting, diarrhea, and colitis, were all mild, with an incidence of grade 3 adverse events <5%. Most treatment-emergent adverse events (TEAEs) occurring during liver and kidney treatment were Grade 1 or 2, with Grade 3 or 4 events occurring in 5% and 5% of patients, respectively. One patient (2%) died within 30 days of treatment initiation; the 60-day mortality rate was 9% (n = 6). Grade 3 mucositis was reported in only 2% of patients. Other non-hematologic adverse events were mild and generally considered unrelated to uproleselan.

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Uproleselan has demonstrated a favorable safety and tolerability profile in both preclinical and clinical studies. In Phase I/II clinical trials (n=19 initial patients), no dose-limiting toxicities (DLTs) were observed when Uproleselan was combined with MEC chemotherapy, leading to the selection of 10 mg/kg intravenous twice daily as the recommended Phase II dose. In relapsed/refractory AML patients (n=47 at RP2D), the most common grade 3/4 treatment-emergent adverse events were associated with chemotherapy itself rather than Uproleselan. Notably, Uproleselan combined with chemotherapy was associated with low rates of oral mucositis, with severe mucositis observed in only 2% of relapsed/refractory patients. This finding is consistent with preclinical observations that Uproleselan protects against chemotherapy-induced mucositis by regulating macrophage trafficking to the site of injury in the gut lining. In a non-human primate venous thrombosis model, Uproleselan monotherapy did not cause clinically significant changes in coagulation parameters, including prothrombin time, activated partial thromboplastin time, and bleeding time, nor were clinically significant bleeding events observed. Platelet counts and fibrinogen levels remained within normal ranges. Collectively, preclinical and clinical data indicate that Uproleselan is well tolerated, does not add cytotoxicity when combined with chemotherapy, and may improve the patient treatment experience by reducing side effects such as mucositis.

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Specific toxicological data for Uproleselan are not detailed in the provided literature. However, in vivo studies have indicated that it can decrease chemotherapy-related toxicity. As a clinical-stage compound, its safety profile is being evaluated in ongoing trials.

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

Uproleselan is a novel, specific E-selectin antagonist currently under investigation in the clinical trial NCT02306291 (Safety, pharmacokinetics, and efficacy study of GMI-1271 in combination with chemotherapy for acute myeloid leukemia). Uproleselan is a synthetic glycomimetic molecule and an E-selectin (CD62E) antagonist with potential antithrombotic, antitumor, and chemotherapeutic effects. After administration, uproleselan binds to E-selectin expressed on the surface of endothelial cells, preventing its interaction with cancer cells expressing selectin E ligands. This may inhibit tumor cell activation, migration, and metastasis. GMI-1271 also interferes with the binding of selectin E-expressing vascular endothelial cells to selectin E-expressing ligands in monocytes and neutrophils, thereby inhibiting their activation. Therefore, this drug inhibits both the activation of the coagulation cascade and thrombus formation. Furthermore, this drug also inhibits leukocyte activation and inflammatory responses. E-selectin is a cell adhesion molecule involved in cell rolling, signal transduction, and chemotaxis; it also plays a key role in inflammatory processes and cancer.

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Uproleselan (CAS 1983970-12-2) is a first-in-class, highly selective small molecule antagonist of E-selectin. It is under investigation for the treatment of acute myeloid leukemia (AML) and has completed Phase II clinical trials with Phase III studies ongoing. Uproleselan is known to have antithrombotic and antineoplastic properties and functions as a tumor microenvironment-disrupting agent.

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.)