TOPK

[1]. TOPK inhibitor induces complete tumor regression in xenograft models of human cancer through inhibition of cytokinesis. Sci Transl Med. 2014 Oct 22;6(259):259ra145.

[2]. Potent anti-myeloma activity of the TOPK inhibitor OTS514 in pre-clinical models. Cancer Med. 2020 Jan;9(1):324-334.

TOPK (T-lymphokine-activated killer cell-derived protein kinase) is highly and frequently activated in various cancer tissues, including lung cancer and triple-negative breast cancer, and plays an indispensable role in cancer cell mitosis. We report the development of a potent TOPK inhibitor, OTS964 {(R)-9-(4-(1-(dimethylamino)propyl-2-yl)phenyl)-8-hydroxy-6-methylthieno[2,3-c]quinoline-4(5H)-one}, which inhibits TOPK kinase activity with high affinity and selectivity. Similar to the knockdown of TOPK small interfering RNA (siRNA), this inhibitor induces cell division defects in vitro and in human lung cancer xenograft models, ultimately inducing apoptosis in cancer cells. Although administration of the free compound induced hematopoietic adverse reactions (leukopenia with thrombocytosis), administration of the liposomal formulation effectively induced complete regression of transplanted tumors in mice without any observed adverse reactions. Our results suggest that inhibiting TOPK activity may be a viable therapeutic approach for treating various human cancers. [1]

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Multiple myeloma (MM) is still considered incurable, thus requiring the development of new drugs. Mitotic kinase T-LAK cell-derived protein kinase/PDZ-binding kinase (TOPK/PBK) is closely related to tumor cell proliferation, cancer stem cell maintenance, and poor patient prognosis in various cancers. In this report, we demonstrate for the first time that the TOPK inhibitor OTS514 has significant anti-myeloma activity. OTS514 can induce cell cycle arrest and apoptosis in a range of human myeloma cell lines (HMCL) at nanomolar concentrations and inhibit the proliferation of putative CD138+ stem cell populations in peripheral blood mononuclear cells of multiple myeloma (MM) patients. In bone marrow cells of MM patients, OTS514 treatment showed stronger killing effects on malignant CD138+ plasma cells than on CD138- plasma cells. In an invasive mouse xenograft model, oral administration of 100 mg/kg OTS964 five days a week was well tolerated, and the tumor volume was reduced by 48%-81% compared with the control group, depending on the initial size of the xenograft. After treatment of HMCL with OTS514, the expression of FOXO3 and its transcriptional targets CDKN1A (p21) and CDKN1B (p27) was upregulated, and apoptosis was induced. TOPK inhibitors can also induce the loss of FOXM1 and disrupt the AKT, p38 MAPK and NF-κB signaling pathways. The effect of OTS514 is independent of p53 mutation or deletion status. The combination of OTS514 and lenalidomide in the treatment of HMCL can produce a synergistic effect, which provides a theoretical basis for evaluating TOPK inhibitors in existing myeloma treatment regimens. [2]

C21H20N2O2S

364.46

364.12

C, 62.91; H, 5.28; Cl, 8.84; N, 6.99; O, 7.98; S, 8.00

1338541-25-5

Typically exists as solid at room temperature

3.25

OETLNMOJNONWOY-LBPRGKRZSA-N

InChI=1S/C21H20N2O2S/c1-11-9-16(24)17(14-5-3-13(4-6-14)12(2)10-22)18-15-7-8-26-20(15)21(25)23-19(11)18/h3-9,12,24H,10,22H2,1-2H3,(H,23,25)/t12-/m0/s1

(R)-9-(4-(1-aminopropan-2-yl)phenyl)-8-hydroxy-6-methylthieno[2,3-c]quinolin-4(5H)-one

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)

DMSO : 31.43 mg/mL (86.24 mM; adjust pH to 3 with 1 M HCL)

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.)