Methods of treating cancer

A technology for cancer and cancer cells, applied in pharmaceutical formulations, medical preparations containing active ingredients, antineoplastic drugs, etc.

Pending Publication Date: 2022-06-07
ASTRAZENECA AB
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, some cancer treatments are only effective for some patients
In addition, some cancer patients develop resistance to certain cancer treatments

Method used

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  • Methods of treating cancer
  • Methods of treating cancer
  • Methods of treating cancer

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0057] Example 1: Development of an FFPE IHC assay specific for SLFN11 and characterization of the DU145SLFN11 KO cell line.

[0058] method

[0059] Knockdown of SLFN11 in DU145 prostate cancer cells by CRISPR / Cas9. Using in-house CRISPR3 software designed to target exon 4 ( The sgRNAS of SLFN11 in the prospacer sequence adjacent motif is bold), synthesized by Integrated DNA Technology (IDT), and cloned into a vector (azPGE02-Cas9-T2A-GFP) containing Cas9 and GFP cassettes . The vector was then transfected into DU145 prostate cancer cells using Lipofectamine 3000 (Thermofisher Scientific). After 48 hours, the pooled single cells with the highest green fluorescent protein (GFP) expression were sorted into 96-well plates. Clones that have lost the wild-type allele are expanded to obtain cell lines from single clones. Two SLFN11-deficient clones were delineated and selected for pharmacological studies (clone KO1 and clone KO2). Cell lysates from SLFN11-perfect (wt) and ...

example 2

[0063] Example 2: Resistance to DDA in DU145 SLFN11 KO cells can be reversed by combination treatment with WEE1 inhibitors.

[0064] method

[0065] Adavotinib was synthesized at AstraZeneca. Gemcitabine, cisplatin, hydroxyurea (HU) and etoposide were obtained from Tocris and camptothecin from Sigma. Stock solutions of gemcitabine (50 mM), cisplatin (1.67 mM) and HU (1 M) were prepared in aqueous solution; all other drugs were dissolved in dimethylsulfoxide (DMSO) (10 mM) at a concentration of 10 mM.

[0066] DU145 isogenic cells (WT and SLFN11 KO) were seeded in 384-well plates and allowed to settle overnight. Figure 2A Immunoblots of SLFN11 WT and KO DU145 isogenic cells used in the experiments are shown. KO 1 and KO 2 are two different CRISPR-KO clones. Cells were dosed with compound solutions (containing the highest doses of 3 μM adavotinib, 0.1 μM gemcitabine and 1 μM etoposide) in a 6x6 concentration matrix using an Echo 555 (LabCyte Corporation). After five cons...

example 3

[0071] Example 3: Resistance to gemcitabine in SLFN11 deficient cell lines can be reversed by combination treatment with WEE1 inhibitors.

[0072] method

[0073] SLFN11 RNA-seq data (log2 RPKM values) were downloaded from Cancer Cell Line Encyclopedia (CCLE) (Barretina J. et al., Nature, 2012;483:603-607), and drug response data (log(IC 50) and the area under the dose-response curve (AUC)) were downloaded from the cancer database drug sensitivity (Yang W et al., Nucleic Acids Res [nucleic acid research], 2013; 41: D955-61). Cell lines with CCLE RNA seq log2 RPKM values ​​below 1 were defined as SLFN11-deficient, and cell lines with log2 RPKM values ​​greater than 1 were defined as SLFN11-perfect. Nineteen pancreatic cell lines in 384-well plates were dosed with increasing concentrations of adavotinib and gemcitabine in a 6x6 concentration matrix using Echo 555 (LabCyte, Inc.). Doses ranged from 0-3 μM for adavotinib and 0-0.3 μM for gemcitabine; in both cases a 1 :3 dilut...

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Abstract

Disclosed herein are methods of treating cancer in a patient with SLFN11 deficient cancer cells with a combination of a WEE1 inhibitor and a DNA damaging agent.

Description

technical field [0001] The present disclosure generally relates to methods of treating cancer. Background technique [0002] WEE1 is a nuclear kinase that belongs to the serine / threonine family of protein kinases. WEE1 inhibits CDKs by phosphorylating cyclin-dependent kinases (CDKs) at two distinct sites (Tyr15 and Thr14). Thus, WEE1 functions in the regulation of mitotic entry and initiation of DNA replication, cell size and the DNA damage checkpoint. Inhibitors of WEE1 have been tested for the treatment of cancer as monotherapy and in combination with other cancer treatments. [0003] Schlafen 11 (SLFN11 ) belongs to the Schlafen protein family and is only expressed in humans and some primates. Inactivation of SLFN11 in cancer cells has been found to lead to resistance to anticancer agents that cause DNA damage and replication stress. Thus, SLFN11 is a determinant of sensitivity to different classes of DNA damaging agents and PARP inhibitors. See Zoppoli et al, PNAS 2...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/06A61K31/519A61P35/00
CPCA61K45/06A61K31/519A61P35/00A61K2300/00A61K31/7068A61K47/6855A61P15/00A61K33/243A61K31/17A61K31/35G01N33/57484G01N2800/52
Inventor E·莱奥C·温克勒M·J·奥康诺尔G·N·琼斯A·J·皮尔斯
Owner ASTRAZENECA AB
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