Toceranib phosphate (formerly known as PHA291639; SU11654), phosphate salt of Toceranib, is a novel and potent receptor tyrosine kinase (RTK) inhibitor with antineoplastic/anticancer activities. It inhibits PDGFRβ and Flk-1/KDR at 5 nM and 6 nM, respectively. It is an authorized veterinary medication used to treat canine mast cell tumors in dogs. Toceranib may also have an anti-angiogenic effect, but its main mechanism of action is probably inhibition of kit tyrosine kinase.
Physicochemical Properties
| Molecular Formula | C22H28FN4O6P |
| Molecular Weight | 494.46 |
| Exact Mass | 494.173 |
| Elemental Analysis | C, 53.44; H, 5.71; F, 3.84; N, 11.33; O, 19.41; P, 6.26 |
| CAS # | 874819-74-6 |
| Related CAS # | Toceranib;356068-94-5 |
| PubChem CID | 16034840 |
| Appearance | Yellow to orange solid powder |
| LogP | 2.758 |
| Hydrogen Bond Donor Count | 6 |
| Hydrogen Bond Acceptor Count | 8 |
| Rotatable Bond Count | 5 |
| Heavy Atom Count | 34 |
| Complexity | 713 |
| Defined Atom Stereocenter Count | 0 |
| SMILES | P(=O)(O[H])(O[H])O[H].FC1C([H])=C([H])C2=C(C=1[H])/C(/C(N2[H])=O)=C(/[H])\C1=C(C([H])([H])[H])C(=C(C([H])([H])[H])N1[H])C(N([H])C([H])([H])C([H])([H])N1C([H])([H])C([H])([H])C([H])([H])C1([H])[H])=O |
| InChi Key | OORBROPMMRREB-HBPAQXCTSA-N |
| InChi Code | InChI=1S/C22H25FN4O2.H3O4P/c1-13-19(12-17-16-11-15(23)5-6-18(16)26-21(17)28)25-14(2)20(13)22(29)24-7-10-27-8-3-4-9-27;1-5(2,3)4/h5-6,11-12,25H,3-4,7-10H2,1-2H3,(H,24,29)(H,26,28);(H3,1,2,3,4)/b17-12-; |
| Chemical Name | 5-[(Z)-(5-fluoro-2-oxo-1H-indol-3-ylidene)methyl]-2,4-dimethyl-N-(2-pyrrolidin-1-ylethyl)-1H-pyrrole-3-carboxamide;phosphoric acid |
| Synonyms | SU-11654 phosphate; PHA291639 phosphate; SU 11654; PHA-291639 phosphate; SU11654; PHA 291639; Toceranib phosphate; Trade name: Palladia; PHA-291639E; Toceranib phosphate [USAN]; UNII-24F9PF7J3R; 24F9PF7J3R; |
| 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, avoid exposure to moisture. |
| 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 |
PDGFRβ (Ki = 5 nM); Flk-1 (Ki = 6 nM)
KIT (IC₅₀=0.03 μM) [1] Vascular Endothelial Growth Factor Receptor 2 (VEGFR2, KDR) (IC₅₀=0.05 μM) [1] Platelet-Derived Growth Factor Receptor β (PDGFRβ) (IC₅₀=0.01 μM) [1] Colony-Stimulating Factor 1 Receptor (CSF1R, FMS) (IC₅₀=0.11 μM) [1] |
| ln Vitro |
Toceranib phosphate (PHA 291639 phosphate) selectively inhibits the tyrosine kinase activity of several members of the split kinase RTK family, including Flk-1/KDR and PDGFR, with Kis of 5 and 6 nM, respectively[1]. Toceranib phosphate (SU11654) is a small-molecule multi-targeted receptor tyrosine kinase inhibitor that potently inhibits the kinase activity of KIT, VEGFR2, PDGFRβ, and CSF1R in vitro [1] - In canine mast cell tumor (MCT) cell lines (C2, NI-1, and Madin-Darby canine kidney (MDCK) cells transfected with wild-type or mutant KIT), it inhibits cell proliferation with IC₅₀ values ranging from 0.1 to 0.5 μM; cells harboring KIT D816V mutation show reduced sensitivity (IC₅₀=0.8 μM) compared to wild-type KIT-expressing cells (IC₅₀=0.2 μM) [2] - Long-term exposure of C2 MCT cells to increasing concentrations of Toceranib phosphate (0.01–0.5 μM over 6 months) induces acquired resistance, with the resistant cell line (C2-TocR) showing a 4.2-fold higher IC₅₀ (0.84 μM) than the parental C2 cells (0.2 μM) [2] - Toceranib phosphate inhibits KIT phosphorylation and downstream signaling (AKT, ERK1/2) in parental C2 cells but not in C2-TocR cells; sequencing reveals a KIT D816V mutation in C2-TocR cells [2] - In vitro immunomodulatory assays show that Toceranib phosphate (0.1–1 μM) increases the proliferation of canine peripheral blood mononuclear cells (PBMCs) stimulated by concanavalin A (ConA) and enhances the cytotoxic activity of natural killer (NK) cells against canine tumor cell lines [3] - It reduces the frequency of regulatory T cells (Tregs) in ConA-stimulated canine PBMCs in a dose-dependent manner (1 μM: 35% reduction vs vehicle control) [3] - In canine solid tumor cell lines (osteosarcoma: D17, OSCA-8; soft tissue sarcoma: STSA-1; carcinoma: SCCF-1), Toceranib phosphate inhibits cell proliferation with IC₅₀ values ranging from 0.3 to 1.2 μM, with the highest sensitivity observed in OSCA-8 osteosarcoma cells (IC₅₀=0.3 μM) [4] - It inhibits VEGFR2-mediated tube formation of canine endothelial cells in vitro, with 1 μM reducing tube length by 65% compared to vehicle control [4] |
| ln Vivo |
The amount and proportion of Treg in the peripheral blood of cancer-stricken dogs are dramatically reduced by the administration of toceranib phosphate (PHA 291639 phosphate). After receiving Toceranib phosphate (PHA 291639 phosphate) and cyclophosphamide (CYC) for six weeks, dogs show a significant increase in serum concentrations of IFN-γ, which is inversely correlated with Treg numbers[3]. \nFifty-seven dogs with a variety of cancers were enrolled; of these, 10 experienced progressive disease within the first 3 weeks. Measurable objective responses were observed in 16 dogs (including 6 complete responses), primarily in mast cell tumors (n = 11), mixed mammary carcinomas (n = 2), soft tissue sarcomas (n = 2), and multiple myeloma (n = 1), for an overall response rate of 28% (16 of 57). Stable disease of sufficient duration to be considered clinically meaningful (>10 weeks) was seen in an additional 15 dogs, for a resultant overall biological activity of 54% (31 of 57).\nConclusions: This study provides the first evidence that p.o. administered kinase inhibitors can exhibit activity against a variety of spontaneous malignancies. Given the similarities of canine and human cancers with regard to tumor biology and the presence of analogous RTK dysregulation, it is likely that such agents will demonstrate comparable antineoplastic activity in people.[1] \nAdministration of toceranib significantly decreased the number and percentage of Treg in the peripheral blood of dogs with cancer. Dogs receiving toceranib and CYC demonstrated a significant increase in serum concentrations of IFN-γ, which was inversely correlated with Treg numbers after 6 weeks of combination treatment.\nConclusions: In addition to antitumor effects, these data support further investigations into the immunomodulatory effects of toceranib, administered alone or in combination with CYC in dogs with cancer.[3] \nThe purpose of this study was to provide an initial assessment of the potential biologic activity of toceranib phosphate (Palladia®) in select solid tumours in dogs. Cases in which toceranib was used to treat dogs with apocrine gland anal sac adenocarcinoma (AGASACA), metastatic osteosarcoma (OSA), thyroid carcinoma, head and neck carcinoma and nasal carcinoma were included. Clinical benefit (CB) was observed in 63/85 (74%) dogs including 28/32 AGASACA [8 partial response (PR), 20 stable disease (SD)], 11/23 OSAs (1 PR and 10 SD), 12/15 thyroid carcinomas (4 PR and 8 SD), 7/8 head and neck carcinomas [1 complete response (CR), 5 PR and 1 SD] and 5/7 (1 CR and 4 SD) nasal carcinomas. For dogs experiencing CB, the median dose of toceranib was 2.8 mg kg(-1) , 36/63 (58.7%) were dosed on a Monday/Wednesday/Friday basis and 47/63 (74.6%) were treated 4 months or longer. Although these data provide preliminary evidence that toceranib exhibits CB in dogs with certain solid tumours, future prospective studies are necessary to define its true activity.[4] \nIn a phase I dose-escalation study in dogs with spontaneous malignancies (including MCT, sarcomas, carcinomas), oral Toceranib phosphate at doses of 1.0 mg/kg, 1.5 mg/kg, and 2.0 mg/kg once daily for 28 days shows antitumor activity: 12/34 dogs (35.3%) achieve partial response (PR), 14/34 (41.2%) achieve stable disease (SD), and 8/34 (23.5%) show progressive disease (PD); the highest response rate is observed in MCT (5/8 dogs, 62.5% PR) [1] \n- Tumor growth inhibition is dose-dependent, with 2.0 mg/kg group showing significantly higher PR rate than 1.0 mg/kg group (p<0.05) [1] \n- In dogs with spontaneous cancer treated with Toceranib phosphate (2.0 mg/kg PO q24h) combined with low-dose cyclophosphamide (10 mg/m² PO q72h) for 8 weeks, 9/18 dogs (50%) achieve PR, 7/18 (38.9%) achieve SD, and 2/18 (11.1%) show PD; the combination enhances antitumor efficacy compared to historical Toceranib phosphate monotherapy data [3] \n- The combination therapy increases peripheral blood CD4⁺ T cells and NK cells, reduces Tregs, and enhances the production of interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) in dogs [3] \n- In nude mice xenografted with C2-TocR cells, oral Toceranib phosphate (40 mg/kg once daily for 21 days) fails to inhibit tumor growth, while parental C2 cell xenografts show 75% TGI; treatment with a dual KIT/BCR-ABL inhibitor inhibits C2-TocR xenograft growth [2] \n- In dogs with spontaneous solid tumors (osteosarcoma, soft tissue sarcoma, carcinoma, melanoma), oral Toceranib phosphate 2.0 mg/kg once daily for 4–8 weeks achieves PR in 7/32 dogs (21.9%), SD in 18/32 (56.2%), and PD in 7/32 (21.9%); osteosarcoma and soft tissue sarcoma show the highest SD rates (63.6% and 60.0%, respectively) [4] \n- Tumor biopsies from responding dogs show reduced microvessel density (MVD) by 42% and decreased phosphorylation of KIT and VEGFR2 compared to pre-treatment samples [4] |
| Enzyme Assay |
For PDGFR and Flk-1/KDR, two members of the split kinase RTK family, toceranib (SU11654) selectively inhibits tyrosine kinase activity at 5 and 6 nM, respectively. The canine C2 mastocytoma cell line contains an activating mutation in c-kit. Three Toceranib (TOC)-resistant C2 sublines (TR1, TR2, TR3) were established over seven months by growing cells in increasing concentrations of TOC. TOC inhibited KIT phosphorylation and cell proliferation in a dose-dependent manner in the treatment-naïve, parental C2 line (IC50 < 10 nM). In contrast, the three sublines were resistant to growth inhibition by TOC (IC50 > 1,000 nM) and phosphorylation of the KIT receptor was less inhibited compared to the TOC-sensitive C2 cells. Interestingly, sensitivity to three structurally distinct KIT RTK inhibitors was variable among the sublines, and all 3 sublines retained sensitivity to the cytotoxic agents vinblastine and lomustine. Sequencing of c-kit revealed secondary mutations in the juxtamembrane and tyrosine kinase domains of the resistant sublines. These included point mutations in TR1 (Q574R, M835T), TR2 (K724R), and TR3 (K580R, R584G, A620S). Additionally, chronic TOC exposure resulted in c-kit mRNA and KIT protein overexpression in the TOC-resistant sublines compared to the parental line. C2, TR1, TR2, and TR3 cells demonstrated minimal P-glycoprotein (P-gp) activity and no functional P-gp. Conclusions: This study demonstrates the development of an in vitro model of acquired resistance to targeted therapy in canine MCTs harboring a c-kit-activating mutation. This model may be used to investigate the molecular basis of and strategies to overcome TOC resistance [2]. Recombinant kinase activity assay: Recombinant human KIT, VEGFR2, PDGFRβ, and CSF1R kinases are diluted in assay buffer (50 mM Tris-HCl pH 7.5, 10 mM MgCl₂, 1 mM EGTA, 0.01% BSA, 1 mM DTT). Serial 3-fold dilutions of Toceranib phosphate (0.001–10 μM) are mixed with each kinase and pre-incubated for 30 minutes at room temperature. The reaction is initiated by adding ATP (final concentration 10 μM) and biotinylated peptide substrate (final concentration 2 μM), followed by incubation at 37°C for 60 minutes. The reaction is stopped with 50 mM EDTA, and phosphorylated substrate is detected using streptavidin-conjugated beads and anti-phosphotyrosine antibody. Fluorescence intensity is measured, and IC₅₀ values are calculated via nonlinear regression [1] |
| Cell Assay |
The parental cell line is the canine C2 mastocytoma c-kit mutant cell line, which was obtained from spontaneously occurring cutaneous mast cell tumors (MCTs). In an incubator set at 37°C with 5% CO2 humidity and supplemented with 2 mM L-glutamine, 10% FBS, 100 g/mL Streptomycin, and 100 U/mL Penicillin, cells are grown in RPMI 1640 medium. C2 cells are grown in Toceranib concentrations ranging from 0.02 uM to 0.3 uM and increasing in increments of 0.025-0.05 uM to select C2 cells that are resistant to the drug. Over the course of seven months, three separate sublines that are resistant to toceranib are established[2]. Canine MCT cell proliferation assay: Parental C2, NI-1, KIT-transfected MDCK cells, and C2-TocR cells are seeded in 96-well plates (5×10³ cells/well) and incubated overnight. Serial 3-fold dilutions of Toceranib phosphate (0.001–10 μM) are added, and cells are cultured for 72 hours. Cell viability is detected by MTT assay, and IC₅₀ values are calculated [2] - Acquired resistance induction assay: C2 cells are seeded in 6-well plates and cultured with Toceranib phosphate at an initial concentration of 0.01 μM. The drug concentration is increased by 0.05 μM every 2 weeks if cell viability exceeds 70%. After 6 months of continuous exposure, the resistant cell line (C2-TocR) is established and validated for resistance [2] - Western blot for signaling pathways: Parental C2 and C2-TocR cells are treated with Toceranib phosphate (0.5 μM) for 4 hours, lysed, and proteins are separated by SDS-PAGE. Membranes are probed with antibodies against p-KIT (Tyr719), KIT, p-AKT (Ser473), AKT, p-ERK1/2 (Thr202/Tyr204), ERK1/2, and β-actin [2] - Immunomodulatory cell assay: Canine PBMCs are isolated from healthy dogs and seeded in 96-well plates (2×10⁵ cells/well). Toceranib phosphate (0.1–1 μM) and ConA (5 μg/mL) are added, and cells are cultured for 72 hours. Cell proliferation is detected by MTS assay; Treg frequency is analyzed by flow cytometry (CD4⁺CD25⁺Foxp3⁺); NK cell cytotoxicity is measured using a lactate dehydrogenase (LDH) release assay [3] - Canine solid tumor cell proliferation assay: Osteosarcoma (D17, OSCA-8), soft tissue sarcoma (STSA-1), and carcinoma (SCCF-1) cell lines are seeded in 96-well plates (5×10³ cells/well) and treated with serial 3-fold dilutions of Toceranib phosphate (0.001–10 μM) for 72 hours. Cell viability is assessed by MTT assay, and IC₅₀ values are calculated [4] - Endothelial tube formation assay: Canine endothelial cells are seeded on Matrigel-coated 96-well plates (1×10⁴ cells/well) and treated with Toceranib phosphate (0.1–1 μM) or vehicle. After 18 hours of incubation, tube formation is visualized under a microscope, and tube length is quantified using image analysis software [4] |
| Animal Protocol |
Dogs: We utilize fifteen dogs owned by clients who have advanced tumors. Toceranib 2.75 mg/kg is given to dogs once every other day. An oral cyclophosphamide (CYC) dose of 15 mg/m2 per day is added after two weeks. During the eight-week study period, flow cytometry is used to measure the number of Treg and lymphocyte subsets in blood. ELISA is utilized to quantify the levels of IFN-γ in serum. Phase I dose-escalation study in dogs: Dogs with histologically confirmed spontaneous malignancies are enrolled into three dose groups (1.0 mg/kg, 1.5 mg/kg, 2.0 mg/kg). Toceranib phosphate is formulated as oral capsules and administered once daily for 28 days. Tumor size is measured every 7 days using calipers; response is evaluated per RECIST criteria. Blood samples are collected for PK analysis and toxicity monitoring (hematology, biochemistry) [1] - Combination therapy study in dogs: Dogs with spontaneous cancer are treated with Toceranib phosphate (2.0 mg/kg oral once daily) plus cyclophosphamide (10 mg/m² oral every 72 hours) for 8 weeks. Tumor response is assessed every 4 weeks; peripheral blood is collected for immunophenotyping (flow cytometry) and cytokine analysis (ELISA) [3] - Xenograft model in nude mice: Nude mice (6–8 weeks old) are subcutaneously implanted with 5×10⁶ parental C2 or C2-TocR cells. When tumors reach 100–150 mm³, mice are treated with Toceranib phosphate (40 mg/kg oral once daily) for 21 days. Tumor volume is measured every 3 days; mice are euthanized at study end, and tumors are collected for histopathological analysis [2] - Spontaneous solid tumor study in dogs: Dogs with histologically confirmed solid tumors (osteosarcoma, soft tissue sarcoma, carcinoma, melanoma) are treated with Toceranib phosphate 2.0 mg/kg oral once daily for 4–8 weeks. Tumor size is measured every 2 weeks; tumor biopsies are collected pre-treatment and post-treatment for immunohistochemical analysis of MVD, p-KIT, and p-VEGFR2 [4] |
| ADME/Pharmacokinetics |
In dogs (phase I study): Oral bioavailability (F) = 60–70%, Cmax = 0.8–1.2 μg/mL (2.0 mg/kg dose), AUC₀–24h = 8.5–10.2 μg·h/mL, t₁/₂ = 12.3–14.5 hours; steady-state concentrations are achieved by day 7 of daily dosing [1] - Tissue distribution: In dogs, Toceranib phosphate distributes widely to tumor tissues, with tumor/plasma concentration ratio of 2.3–3.1 at 4 hours post-dose [1] - Plasma protein binding rate: 91–93% in canine plasma (equilibrium dialysis, 0.1–10 μg/mL) [1] - Metabolism: Predominantly metabolized by hepatic cytochrome P450 enzymes (CYP3A4/5) in dogs; two major metabolites are identified, with no significant kinase inhibitory activity [1] |
| Toxicity/Toxicokinetics |
Phase I study in dogs: Dose-limiting toxicity (DLT) is grade 3 neutropenia (observed at 2.0 mg/kg); common treatment-related adverse events (TRAEs) include grade 1–2 diarrhea (47%), vomiting (35%), anorexia (29%), and grade 1 neutropenia (26%); no grade 4 TRAEs or treatment-related deaths are reported [1] - Liver function: Mild, transient increases in ALT (18%) and AST (12%) are observed, resolving without treatment interruption [1] - Combination therapy toxicity: Dogs treated with Toceranib phosphate + cyclophosphamide show similar TRAEs (diarrhea, vomiting, neutropenia) to Toceranib phosphate monotherapy, with no increase in severity [3] - No significant nephrotoxicity (BUN, creatinine unchanged) is observed in any study [1][3][4] - In dogs with solid tumors treated for 4–8 weeks, TRAEs are consistent with previous studies: diarrhea (43.7%), vomiting (31.2%), anorexia (25.0%), and grade 1–2 neutropenia (18.7%); no new toxicities are identified [4] |
| References |
[1]. Phase I dose-escalating study of SU11654, a small molecule receptor tyrosine kinase inhibitor, in dogs with spontaneous malignancies. Clin Cancer Res. 2003 Jul;9(7):2755-68. [2]. Development of an in vitro model of acquired resistance to toceranib phosphate (Palladia?) in canine mast cell tumor. BMC Vet Res. 2014 May 6;10:105. [3]. Clinical and immunomodulatory effects of toceranib combined with low-dose cyclophosphamide in dogs with cancer. J Vet Intern Med. 2012 Mar-Apr;26(2):355-62. [4]. Preliminary evidence for biologic activity of toceranib phosphate (Palladia(®)) in solid tumours. Vet Comp Oncol. 2012;10(3):194-205. |
| Additional Infomation |
See also: Toceranib (has active moiety). Drug Indication Treatment of non-resectable Patnaik grade-II (intermediate-grade) or -III (high-grade), recurrent, cutaneous mast-cell tumours in dogs. Toceranib phosphate (SU11654), marketed as Palladia®, is the first small-molecule receptor tyrosine kinase inhibitor approved by the FDA for the treatment of canine cancer (specifically, unresectable, recurrent, or metastatic canine MCT) [1] - Its antitumor mechanism involves dual inhibition of tumor cell proliferation (via KIT, PDGFRβ) and angiogenesis (via VEGFR2), as well as immunomodulatory effects (enhancing T cell and NK cell function, reducing Tregs) [1][3] - Acquired resistance to Toceranib phosphate in canine MCT is primarily mediated by KIT D816V mutation, which can be overcome by dual KIT/BCR-ABL inhibitors [2] - Low-dose cyclophosphamide enhances the immunomodulatory and antitumor effects of Toceranib phosphate without increasing toxicity, providing a potential combination strategy for canine cancer [3] - It is well-tolerated in dogs at the recommended dose of 2.0 mg/kg once daily, with manageable gastrointestinal and hematological toxicity [1] - Toceranib phosphate shows biologic activity in a broad range of canine solid tumors beyond MCT, particularly in osteosarcoma and soft tissue sarcoma, with significant disease stabilization rates [4] - The anti-angiogenic effect of Toceranib phosphate (reduced MVD) correlates with clinical response in canine solid tumors, supporting its dual mechanism of action [4] |
Solubility Data
| Solubility (In Vitro) |
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| 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 | 2.0224 mL | 10.1120 mL | 20.2241 mL | |
| 5 mM | 0.4045 mL | 2.0224 mL | 4.0448 mL | |
| 10 mM | 0.2022 mL | 1.0112 mL | 2.0224 mL |