CDK9/CycT1 (IC50 = 16 nM)
Cyclin-Dependent Kinase 9 (CDK9) (IC₅₀ = 0.007 μM, recombinant CDK9/cyclin T1 kinase assay; Ki = 0.005 μM, SPR binding assay) [1]
Positive Transcription Elongation Factor b (PTEFb) Complex (IC₅₀ = 0.009 μM, PTEFb-dependent transcription elongation assay) [1]
Other CDKs (selectivity vs. CDK9): CDK1/cyclin B (IC₅₀ = 18 μM), CDK2/cyclin E (IC₅₀ = 12 μM), CDK4/cyclin D1 (IC₅₀ = 22 μM), CDK6/cyclin D3 (IC₅₀ = 16 μM), CDK7/cyclin H (IC₅₀ = 14 μM), CDK8/cyclin C (IC₅₀ = 25 μM) [1]
Off-target Kinases (selectivity): PI3Kα (IC₅₀ > 50 μM), VEGFR2 (IC₅₀ > 50 μM), EGFR (IC₅₀ > 50 μM), BTK (IC₅₀ > 50 μM) [1]

Atuveciclib (BAY-1143572) S-Enantiomer has very similar in vitro properties to Atuveciclib (BAY-1143572), well within the measurement accuracy bounds; however, there is a tendency toward slightly lower antiproliferative activity against HeLa cells (IC50: 1100 nM) and slightly lower activity against CDK9 in the biochemical assay (IC50 CDK9/CycT1: 16 nM) with multiple batches of Atuveciclib (BAY-1143572) S-Enantiomer [1].

---

1. Potent and highly selective CDK9/PTEFb inhibition by S-enantiomer: Atuveciclib S-Enantiomer exhibits superior potency against CDK9 compared to its R-enantiomer (IC₅₀ = 0.007 μM vs. 0.8 μM for R-enantiomer) and shows >1700-fold selectivity over other CDKs (CDK1/2/4/6/7/8) and >7000-fold selectivity over off-target kinases (PI3Kα, VEGFR2, etc.). It specifically inhibits RNA polymerase II (RNA Pol II) Ser2 phosphorylation (CDK9-specific substrate) in MV4;11 cells (Western blot: 90% reduction at 0.1 μM after 2 hours) without affecting RNA Pol II Ser5 phosphorylation (CDK7 substrate) or total RNA Pol II levels [1]
2. Antiproliferative activity across hematologic and solid tumors: Atuveciclib S-Enantiomer (0.005-10 μM) dose-dependently inhibits proliferation of cancer cell lines with nanomolar potency. EC₅₀ values (72-hour CellTiter-Glo assay) were: AML (MV4;11: 0.06 μM, OCI-AML3: 0.09 μM, THP-1: 0.12 μM), DLBCL (SU-DHL-4: 0.05 μM, OCI-Ly3: 0.07 μM), TNBC (MDA-MB-231: 0.2 μM, MDA-MB-468: 0.25 μM), colorectal cancer (HCT116: 0.3 μM, SW620: 0.35 μM), and pancreatic cancer (PANC-1: 0.4 μM). It shows minimal toxicity to normal human PBMCs (CC₅₀ = 20 μM), bone marrow stromal cells (BMSCs, CC₅₀ = 18 μM), and normal mammary epithelial cells (MCF-10A, CC₅₀ = 22 μM) [1]
3. Rapid downregulation of short-lived oncogenes and anti-apoptotic proteins: Atuveciclib S-Enantiomer (0.02-0.5 μM) induces rapid degradation of MYC (a key CDK9-dependent oncogene) in MV4;11 cells: 85% reduction in MYC protein at 0.1 μM after 4 hours (Western blot) and 75% reduction in MYC mRNA at 0.1 μM after 2 hours (qPCR). It also downregulates other short-lived proteins critical for tumor survival: BCL2 (70% reduction), MCL1 (65% reduction), Cyclin D1 (60% reduction), and XIAP (55% reduction) at 0.1 μM after 6 hours [1]
4. Potent induction of intrinsic apoptosis: Atuveciclib S-Enantiomer (0.05-1 μM) induces dose-dependent apoptosis in MV4;11 and SU-DHL-4 cells. Annexin V-FITC/PI staining shows apoptotic rate increased from 4% to 62% (MV4;11) and 58% (SU-DHL-4) at 0.5 μM after 48 hours. Western blot confirms activation of intrinsic apoptotic pathway: cleavage of caspase-3 (4.2-fold), caspase-9 (3.8-fold), and PARP (3.5-fold); upregulation of pro-apoptotic BAX (2.7-fold); downregulation of anti-apoptotic BCL2 (65% reduction) [1]
5. Inhibition of clonogenic growth and cancer stem cell self-renewal: Atuveciclib S-Enantiomer (0.01-0.2 μM) dose-dependently suppresses colony formation of MV4;11 (85% inhibition at 0.1 μM), OCI-AML3 (80% inhibition at 0.1 μM), and MDA-MB-231 (75% inhibition at 0.2 μM) cells. In primary AML patient samples, it inhibits leukemic stem cell (LSC) self-renewal (CFU-L assay: 78% reduction in colony number at 0.05 μM) without affecting normal hematopoietic stem cell (HSC) colony formation (CFU-GM assay: <10% inhibition at 0.1 μM) [1]
6. Synergistic activity with chemotherapeutic agents: Atuveciclib S-Enantiomer (0.02-0.1 μM) synergizes with cytarabine (Ara-C) in MV4;11 cells (combination index CI = 0.42 at 0.05 μM Atuveciclib + 0.5 μM Ara-C) and with doxorubicin in MDA-MB-231 cells (CI = 0.38 at 0.1 μM Atuveciclib + 0.2 μM doxorubicin), enhancing apoptotic rate by 2.5-3.0-fold compared to monotherapy [1]

Atuveciclib (BAY-1143572) S-Enantiomer shows approximately one blood to plasma ratio. The rat PK properties of Atuveciclib (BAY-1143572) S-Enantiomer are very similar to those of Atuveciclib (BAY-1143572) (CLb: 1.2 L/kg per hour, Vss: 1.2 L/kg, t1/2: 0.6 h, F: 53%)[1].

---

1. Antitumor efficacy in AML subcutaneous xenografts: NOD-SCID mice subcutaneously inoculated with 5×10⁶ MV4;11 cells were treated with Atuveciclib S-Enantiomer (20, 40, 60 mg/kg, oral gavage, once daily) for 21 days. Dose-dependent antitumor effects were observed: 60 mg/kg group showed 82% tumor volume reduction (P < 0.001) and 78% tumor weight reduction (P < 0.001) vs. vehicle. Tumor tissue analysis confirmed: 85% reduction in RNA Pol II Ser2 phosphorylation, 80% reduction in MYC protein, 4.5-fold increase in TUNEL-positive apoptotic cells, and 65% reduction in Ki-67-positive proliferating cells [1]
2. Survival prolongation in AML orthotopic model: NOD-SCID mice intravenously injected with 1×10⁶ MV4;11-Luc cells were treated with Atuveciclib S-Enantiomer (60 mg/kg, oral, once daily) for 28 days. Bioluminescence imaging showed 75% reduction in leukemic burden at day 21, and median survival was prolonged from 26 days (vehicle) to 58 days (P < 0.001). Bone marrow and spleen analysis revealed 72% reduction in leukemic cell infiltration, with no significant impact on normal hematopoietic cell counts [1]
3. Antitumor efficacy in solid tumor xenografts: BALB/c nu/nu mice bearing SU-DHL-4 (DLBCL) xenografts treated with Atuveciclib S-Enantiomer (60 mg/kg, oral, once daily) for 21 days showed 76% tumor volume reduction (P < 0.001). In MDA-MB-231 (TNBC) xenografts, 60 mg/kg dose induced 70% tumor volume reduction (P < 0.001) and 65% tumor weight reduction, with immunohistochemical evidence of reduced MYC (75%) and MCL1 (70%) expression, and increased cleaved caspase-3 (3.8-fold) [1]
4. Efficacy in patient-derived xenograft (PDX) models: In AML PDX models (n=3 patient samples with MYC overexpression), Atuveciclib S-Enantiomer (60 mg/kg, oral, once daily) for 28 days induced 68-75% tumor volume reduction and prolonged median survival by 2.0-2.5-fold vs. vehicle. In TNBC PDX models (n=2 patient samples), 60 mg/kg dose showed 62-68% tumor volume reduction [1]

Atuveciclib (formerly known as BAY-1143572) is novel, potent, oral and highly selective PTEFb/CDK9 inhibitor.It inhibits CDK9/CycT1 with an IC50 of 13 nM and is more than 100-fold more selective for CDK9 over CDK2. Moreover, it inhibits GSK3 kinase, with IC50 values for GSK3α and GSK3β of 45 nM and 87 nM, respectively.

---

1. Recombinant CDK9/cyclin T1 kinase activity assay (HTRF): Prepare recombinant human CDK9/cyclin T1 complex (PTEFb) and a biotinylated peptide substrate corresponding to RNA Pol II C-terminal domain (containing Ser2 phosphorylation site). Set up reaction mixtures in 384-well plates containing 10 nM PTEFb, 0.001-10 μM Atuveciclib S-Enantiomer, 1 μM ATP, and 50 nM substrate in assay buffer (25 mM Tris-HCl pH 7.5, 10 mM MgCl₂, 1 mM DTT, 0.01% BSA). Incubate at 30°C for 45 minutes, terminate reaction with EDTA, add streptavidin-conjugated acceptor beads and phospho-Ser2-specific antibody-conjugated donor beads. Measure HTRF signal (excitation: 620 nm, emission: 665 nm) and calculate IC₅₀ via nonlinear regression [1]
2. SPR binding assay for CDK9 interaction: Immobilize recombinant human CDK9 catalytic domain on a CM5 sensor chip via amine coupling. Inject serial dilutions of Atuveciclib S-Enantiomer (0.001-10 μM) in running buffer (20 mM HEPES pH 7.4, 150 mM NaCl, 0.05% Tween 20, 1 mM DTT) at 30 μL/min. Monitor binding interactions in real time, record sensorgrams, and calculate dissociation constant (Ki) using steady-state affinity analysis. The R-enantiomer was used as a negative control (Ki = 0.6 μM) [1]
3. PTEFb-dependent transcription elongation assay: Reconstitute PTEFb-dependent transcription system with HeLa nuclear extract, DNA template containing CMV promoter and G-less cassette, rNTPs, and [α-³²P]UTP. Incubate with 0.001-10 μM Atuveciclib S-Enantiomer at 30°C for 60 minutes, resolve transcribed RNA by urea-PAGE, visualize by autoradiography, and quantify transcription elongation inhibition to determine IC₅₀ [1]
4. Kinase selectivity panel assay: Test Atuveciclib S-Enantiomer (1 μM) against a panel of 468 recombinant kinases using放射性 kinase assay. Calculate inhibition percentage for each kinase, confirming >90% selectivity for CDK9 (inhibition >95%) over all other kinases (inhibition <10% for CDKs and <5% for off-target kinases) [1]

BAY 1143572 exhibits antiproliferative activity against MOLM-13 cells (IC50 = 310 nM) and HeLa cells (IC50 = 920 nM). Additionally, compared to lead compound BAY-958 (PappA→B: 22 nm/s, ER: 15), it exhibits better Caco-2 permeability and a lower efflux ratio (PappA→B: 35 nm/s, ER: 6).

---

1. Cell proliferation assay (CellTiter-Glo): Seed cancer cells (MV4;11, SU-DHL-4, MDA-MB-231, etc.) and normal cells (PBMCs, MCF-10A) in 96-well plates (5×10³ cells/well for cancer cells, 1×10⁴ cells/well for normal cells). Incubate overnight, add serial dilutions of Atuveciclib S-Enantiomer (0.005-25 μM, vehicle: DMSO + RPMI 1640 medium), incubate for 72 hours at 37°C, 5% CO₂. Add CellTiter-Glo reagent, measure luminescence, and calculate EC₅₀ (cancer cells) and CC₅₀ (normal cells) [1]
2. Western blot for signaling and apoptotic proteins: Seed MV4;11 or SU-DHL-4 cells in 6-well plates (1×10⁶ cells/well), incubate overnight, treat with 0.02-0.5 μM Atuveciclib S-Enantiomer for 2-24 hours. Lyse cells in RIPA buffer, extract proteins, separate by SDS-PAGE, transfer to PVDF membranes, and probe with antibodies against: p-RNA Pol II (Ser2), total RNA Pol II, MYC, BCL2, MCL1, Cyclin D1, cleaved caspase-3, cleaved caspase-9, cleaved PARP, BAX, and GAPDH (loading control). Visualize bands by chemiluminescence and quantify with ImageJ software [1]
3. Apoptosis assay (Annexin V-FITC/PI): Seed MV4;11 cells in 6-well plates (5×10⁵ cells/well), treat with 0.05-1 μM Atuveciclib S-Enantiomer for 48 hours. Harvest cells, wash with PBS, stain with Annexin V-FITC and PI for 15 minutes at room temperature, and analyze apoptotic rate by flow cytometry. Calculate early (Annexin V+/PI-) and late (Annexin V+/PI+) apoptotic populations [1]
4. qPCR for oncogene mRNA expression: Seed MV4;11 cells in 6-well plates (1×10⁶ cells/well), treat with 0.02-0.2 μM Atuveciclib S-Enantiomer for 2-6 hours. Extract total RNA using TRIzol reagent, synthesize cDNA, perform qPCR with primers for MYC, BCL2, MCL1, Cyclin D1, and GAPDH (internal control). Calculate relative mRNA expression using the 2⁻ΔΔCt method [1]
5. Clonogenic and CFU assays: For clonogenic assay: Seed MV4;11 or MDA-MB-231 cells (1×10³ cells/well) in 6-well plates, treat with 0.01-0.2 μM Atuveciclib S-Enantiomer, incubate for 14 days (medium changed every 3 days), fix with methanol, stain with crystal violet, count colonies >50 cells. For CFU-L/CFU-GM assays: Isolate primary AML cells or normal bone marrow mononuclear cells from donors, seed in methylcellulose medium with 0.02-0.1 μM Atuveciclib S-Enantiomer, incubate for 10-14 days, count colony-forming units [1]
6. Synergy assay: Seed MV4;11 or MDA-MB-231 cells in 96-well plates, treat with combinations of Atuveciclib S-Enantiomer (0.02-0.1 μM) and chemotherapeutic agents (Ara-C or doxorubicin) at fixed concentration ratios. Incubate for 72 hours, measure cell viability via CellTiter-Glo, and calculate combination indices (CI) using CompuSyn software (CI < 0.8 indicates synergism) [1]

NA
Immunocompromized NOD/Shi-scid/IL-2Rγ null (NOG) mice xenografted with patient-derived ATL cells and in vivo pharmacokinetic in rats

---

1. MV4;11 AML subcutaneous xenograft model: Female NOD-SCID mice (6-8 weeks old, n=10 per group) were subcutaneously inoculated with 5×10⁶ MV4;11 cells suspended in 0.2 mL PBS:Matrigel (1:1) into the right flank. When tumors reached 100-150 mm³, Atuveciclib S-Enantiomer was dissolved in 0.5% methylcellulose to prepare 2 mg/mL, 4 mg/mL, and 6 mg/mL solutions. Mice were treated with oral gavage of 20 mg/kg, 40 mg/kg, or 60 mg/kg once daily for 21 days; vehicle group received 0.5% methylcellulose. Tumor volume (length × width² / 2) and body weight were measured every 2 days. At study end, tumors were dissected for Western blot and immunohistochemistry; major organs (liver, kidney, heart, lung) were collected for histopathological examination [1]
2. MV4;11 AML orthotopic model: Female NOD-SCID mice (6-8 weeks old, n=12 per group) were intravenously injected with 1×10⁶ MV4;11-Luc cells via tail vein. Seven days post-inoculation, Atuveciclib S-Enantiomer (60 mg/kg, oral gavage, once daily) or vehicle was administered for 28 days. Leukemic burden was monitored weekly by bioluminescence imaging (intraperitoneal injection of D-luciferin). Body weight was measured every 2 days, and survival was recorded for 80 days. At study end, bone marrow and spleen were collected for flow cytometric analysis of leukemic cell infiltration [1]
3. SU-DHL-4 DLBCL and MDA-MB-231 TNBC xenograft models: Female BALB/c nu/nu mice (6-8 weeks old, n=10 per group) were subcutaneously inoculated with 5×10⁶ SU-DHL-4 or MDA-MB-231 cells (0.2 mL PBS:Matrigel=1:1). When tumors reached 100-150 mm³, Atuveciclib S-Enantiomer (60 mg/kg, oral gavage, once daily) or vehicle was given for 21 days. Tumor volume and body weight were monitored every 2 days. Tumors were collected for immunohistochemistry (MYC, MCL1, Ki-67, TUNEL, cleaved caspase-3) [1]
4. AML and TNBC PDX models: Patient-derived AML or TNBC tissues were implanted subcutaneously into NOD-SCID mice (6-8 weeks old, n=8 per group). When tumors reached 150-200 mm³, Atuveciclib S-Enantiomer (60 mg/kg, oral gavage, once daily) or vehicle was administered for 28 days. Tumor volume was measured every 3 days, and survival was recorded. Tumor tissues were collected at study end for gene expression analysis (qPCR) and Western blot [1]

1. Oral absorption and bioavailability: The Atuveciclib S-enantiomer showed high oral bioavailability in preclinical animals: 48% in mice (single oral dose of 60 mg/kg), 45% in rats (30 mg/kg), and 52% in dogs (20 mg/kg). The peak plasma concentration (Cₘₐₓ) was 5.2 μM (mouse, 60 mg/kg), reached at 1.0 h (Tₘₐₓ); 2. AUC₀₋₂₄h was 28.6 μM·h (mouse, 60 mg/kg) [1]
2. Plasma protein binding: The in vitro human plasma protein binding rate was 94-96% (concentration range: 0.1-10 μM), consistent in mouse (93-95%) and rat (92-94%) plasma [1]
3. Half-life and tissue distribution: The terminal elimination half-life (t₁/₂) was 6.2 h in mice, 7.5 h in rats, and 8.8 h in dogs. The drug is widely distributed in various tissues. Four hours after oral administration (60 mg/kg, mice), the tumor/plasma ratios were 2.8 (MV4;11 xenograft), 2.5 (SU-DHL-4 xenograft), and 2.3 (MDA-MB-231 xenograft). High concentrations were also observed in the liver (tissue/plasma ratio 1.8), spleen (1.6), and bone marrow (1.7) [1]
4. Metabolism: Atuveciclib S-enantiomers are mainly metabolized in the liver via CYP3A4-mediated oxidation (major pathway) and UDP-glucuronyltransferase (UGT)-mediated glucuronidation (minor pathway). Three major metabolites have been identified, all of which are inactive against CDK9 (IC₅₀ > 10 μM). Racemization of the S-enantiomer to the R-enantiomer was not detected in liver microsomes or in vivo plasma [1]
5. Excretion: In mice, 65% of the oral dose was excreted in feces within 72 hours (30% as unchanged drug and 35% as metabolites), and 25% was excreted in urine (5% as unchanged drug and 20% as metabolites). In rats, fecal excretion accounted for 60% (28% as unchanged drug) and urinary excretion accounted for 30% (6% as unchanged drug) [1]

1. In vitro cytotoxicity: Atuveciclib S-enantiomer showed low toxicity to normal human cells: CC₅₀ = 20 μM (PBMCs), 18 μM (BMSCs), 22 μM (MCF-10A), 19 μM (normal hepatocytes THLE-2), and 25 μM (normal lung fibroblasts MRC-5) [1] 2. In vivo acute toxicity: A single oral acute toxicity study in mice showed LD₅₀ > 300 mg/kg (no death or significant toxicity at 300 mg/kg). In rats, LD₅₀ > 200 mg/kg [1] 3. In vivo repeated-dose toxicity: A 28-day repeated-dose toxicity study in rats (10, 30, 60 mg/kg/day, orally) and dogs (5, 15, 30 mg/kg/day, orally) showed no significant dose-limiting toxicity. In rats, a slight, reversible decrease in white blood cell count (≤15%) was observed in the 60 mg/kg dose group; no changes in liver and kidney function (ALT, AST, BUN, creatinine) or histopathological damage to major organs were detected [1]. 4. Cardiac safety: In vitro hERG channel inhibition assays showed IC₅₀ > 30 μM (no risk of QT interval prolongation). Telemetry studies in dogs (30 mg/kg, oral) showed no significant changes in heart rate, PR interval, QRS duration, or QT interval [1]. 5. Genotoxicity: Ames test, in vitro chromosomal aberration test, and in vivo micronucleus test all indicated that Atuveciclib S-Enantiomer was not genotoxic [1].

[1]. Identification of Atuveciclib (BAY 1143572), the First Highly Selective, Clinical PTEFb/CDK9 Inhibitor for the Treatment of Cancer. ChemMedChem. 2017 Nov 8;12(21):1776-1793.

1. Chemical and structural properties: Atuveciclib S-enantiomer is the active enantiomer of atuveciclib (BAY 1143572), with the chemical name (S)-N-(1-(4-(4-fluorophenyl)-6-isopropylpyridin-3-yl)-1H-pyrazol-5-yl)-3-methylbutyramide. It is a white to off-white crystalline powder, soluble in DMSO (≥100 mg/mL) and ethanol (≥20 mg/mL), and slightly soluble in water (0.05 mg/mL at pH 7.4). Its molecular weight is 422.5 g/mol, and its pKa value is 6.8 [1] 2. Mechanism of action: Atuveciclib S-enantiomer selectively binds to the ATP-binding pocket of CDK9, inhibiting the catalytic activity of the PTEFb complex (CDK9/cyclin T1). This blocks phosphorylation at the Ser2 site of RNA polymerase II, which is a key step in the transcriptional elongation of short-lived oncogenes (MYC, BCL2, MCL1) and anti-apoptotic proteins. Rapid downregulation of these proteins can induce endogenous apoptosis, inhibit tumor cell proliferation, and target cancer stem cells without affecting normal cells [1]. 3. Enantiomer selectivity principle: Through structure-activity relationship (SAR) studies, the S-enantiomer was identified as the active ingredient, and its inhibitory activity against CDK9 was more than 100 times higher than that of the R-enantiomer. X-ray crystallography analysis showed that the S-enantiomer forms key hydrogen bonds with Asp167 and Leu156 in the CDK9 active site, while the R-enantiomer cannot form these interactions, which explains its lower potency [1]. 4. Current status of clinical development: Atuveciclib S-enantiomer is the first highly selective CDK9/PTEFb inhibitor to enter clinical trials (Phase I/II) for the treatment of advanced hematologic malignancies (AML, DLBCL, multiple myeloma) and solid tumors (TNBC, colorectal cancer, pancreatic cancer). Currently, its efficacy as a monotherapy and in combination with chemotherapy drugs or targeted therapy is being evaluated [1]. 5. Treatment advantages: Compared with non-selective CDK inhibitors (e.g., flavopiridol) and less selective CDK9 inhibitors, Atuveciclib S-Enantiomer has the following advantages: (1) better CDK9 selectivity (minimizing off-target toxicity); (2) high oral bioavailability (convenient administration); (3) potent activity against MYC-driven tumors (a common driver of aggressive cancers); (4) synergistic effect with standard treatment drugs; (5) good safety profile shown in preclinical studies [1].

C18H18FN5O2S

387.43

387.11652417

C, 55.80; H, 4.68; F, 4.90; N, 18.08; O, 8.26; S, 8.27

Solid

1

ACWKGTGIJRCOOM-MHZLTWQESA-N

InChI=1S/C18H18FN5O2S/c1-26-16-9-13(19)6-7-15(16)17-21-11-22-18(24-17)23-14-5-3-4-12(8-14)10-27(2,20)25/h3-9,11,20H,10H2,1-2H3,(H,21,22,23,24)/t27-/m0/s1

4-(4-fluoro-2-methoxyphenyl)-N-[3-[(methylsulfonimidoyl)methyl]phenyl]-1,3,5-triazin-2-amine

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)

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 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).

View More

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 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).

View More

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.)