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Target modulation by a kinase inhibitor engineered to induce a tandem blockade of the epidermal growth factor receptor (EGFR) and c-Src: the concept of type III combi-targeting.

Rao S, Larroque-Lombard AL, Peyrard L, Thauvin C, Rachid Z, Williams C, Jean-Claude BJ - PLoS ONE (2015)

Bottom Line: Cancer cells are characterized by a complex network of interrelated and compensatory signaling driven by multiple kinases that reduce their sensitivity to targeted therapy.Variation of K1-K2 linkers led to AL776, our first optimized EGFR-c-Src targeting prototype.We now term this new targeting model consisting of designing a kinase inhibitor K1-K2 to target Kin-1 and Kin-2, and to further release two inhibitors K1 and K2 of the latter kinases, "type III combi-targeting".

View Article: PubMed Central - PubMed

Affiliation: Cancer Drug Research Laboratory, Department of Medicine, Division of Medical Oncology, McGill University Health Center/Royal Victoria Hospital, 687 Pine Avenue West Rm M7.19, Montreal, Quebec, H3A 1A1 Canada.

ABSTRACT
Cancer cells are characterized by a complex network of interrelated and compensatory signaling driven by multiple kinases that reduce their sensitivity to targeted therapy. Therefore, strategies directed at inhibiting two or more kinases are required to robustly block the growth of refractory tumour cells. Here we report on a novel strategy to promote sustained inhibition of two oncogenic kinases (Kin-1 and Kin-2) by designing a molecule K1-K2, termed "combi-molecule", to induce a tandem blockade of Kin-1 and Kin-2, as an intact structure and to be further hydrolyzed to two inhibitors K1 and K2 directed at Kin-1 and Kin-2, respectively. We chose to target EGFR (Kin-1) and c-Src (Kin-2), two tyrosine kinases known to synergize to promote tumour growth and progression. Variation of K1-K2 linkers led to AL776, our first optimized EGFR-c-Src targeting prototype. Here we showed that: (a) AL776 blocked EGFR and c-Src as an intact structure using an in vitro kinase assay (IC50 EGFR = 0.12 μM and IC50 c-Src = 3 nM), (b) it could release K1 (AL621, a nanomolar EGFR inhibitor) and K2 (dasatinib, a clinically approved Abl/c-Src inhibitor) by hydrolytic cleavage both in vitro and in vivo, (c) it could robustly inhibit phosphorylation of EGFR and c-Src (0.25-1 μM) in cells, (d) it induced 2-4 fold stronger growth inhibition than gefitinib or dasatinib and apoptosis at concentrations as low as 1 μM, and, (e) blocked motility and invasion at sub-micromolar doses in the highly invasive 4T1 and MDA-MB-231 cells. Despite its size (MW = 1032), AL776 blocked phosphorylation of EGFR and c-Src in 4T1 tumours in vivo. We now term this new targeting model consisting of designing a kinase inhibitor K1-K2 to target Kin-1 and Kin-2, and to further release two inhibitors K1 and K2 of the latter kinases, "type III combi-targeting".

No MeSH data available.


Related in: MedlinePlus

Synthesis and hydrolysis of AL776, the lead K1-K2 prototype targetingEGFR and c-Src.The synthesis of AL776 was carried out in our laboratory according to thesteps indicated above. The resulting type III K1-K2 molecule is designedto undergo hydrolysis inside the cells and release a potent EGFRtyrosine kinase inhibitor (K1) termed AL621 and a potent c-Src tyrosinekinase inhibitor (K2) dasatinib (type I). AL776 is also capable ofexerting its dual inhibitory property by directly interacting with eachtarget as an intact molecule (type II).
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pone.0117215.g003: Synthesis and hydrolysis of AL776, the lead K1-K2 prototype targetingEGFR and c-Src.The synthesis of AL776 was carried out in our laboratory according to thesteps indicated above. The resulting type III K1-K2 molecule is designedto undergo hydrolysis inside the cells and release a potent EGFRtyrosine kinase inhibitor (K1) termed AL621 and a potent c-Src tyrosinekinase inhibitor (K2) dasatinib (type I). AL776 is also capable ofexerting its dual inhibitory property by directly interacting with eachtarget as an intact molecule (type II).

Mentions: The synthesis of AL776 proceeded according to Fig. 3. Dasatinib was treated with an excess ofsuccinic anhydride to give compound 1, which was coupled with AL621(a potent EGFR tyrosine kinase inhibitor with IC50 = 3 nM [12]) in the presence ofEDCI, HOBt and DMAP to give VII (AL776) as an analytically purewhite powder following purification by preparative TLC. We predicted that thehydrolysis of AL776 would restore its primary synthetic elements (i.e. AL621 asK1 and dasatinib as K2).


Target modulation by a kinase inhibitor engineered to induce a tandem blockade of the epidermal growth factor receptor (EGFR) and c-Src: the concept of type III combi-targeting.

Rao S, Larroque-Lombard AL, Peyrard L, Thauvin C, Rachid Z, Williams C, Jean-Claude BJ - PLoS ONE (2015)

Synthesis and hydrolysis of AL776, the lead K1-K2 prototype targetingEGFR and c-Src.The synthesis of AL776 was carried out in our laboratory according to thesteps indicated above. The resulting type III K1-K2 molecule is designedto undergo hydrolysis inside the cells and release a potent EGFRtyrosine kinase inhibitor (K1) termed AL621 and a potent c-Src tyrosinekinase inhibitor (K2) dasatinib (type I). AL776 is also capable ofexerting its dual inhibitory property by directly interacting with eachtarget as an intact molecule (type II).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4414309&req=5

pone.0117215.g003: Synthesis and hydrolysis of AL776, the lead K1-K2 prototype targetingEGFR and c-Src.The synthesis of AL776 was carried out in our laboratory according to thesteps indicated above. The resulting type III K1-K2 molecule is designedto undergo hydrolysis inside the cells and release a potent EGFRtyrosine kinase inhibitor (K1) termed AL621 and a potent c-Src tyrosinekinase inhibitor (K2) dasatinib (type I). AL776 is also capable ofexerting its dual inhibitory property by directly interacting with eachtarget as an intact molecule (type II).
Mentions: The synthesis of AL776 proceeded according to Fig. 3. Dasatinib was treated with an excess ofsuccinic anhydride to give compound 1, which was coupled with AL621(a potent EGFR tyrosine kinase inhibitor with IC50 = 3 nM [12]) in the presence ofEDCI, HOBt and DMAP to give VII (AL776) as an analytically purewhite powder following purification by preparative TLC. We predicted that thehydrolysis of AL776 would restore its primary synthetic elements (i.e. AL621 asK1 and dasatinib as K2).

Bottom Line: Cancer cells are characterized by a complex network of interrelated and compensatory signaling driven by multiple kinases that reduce their sensitivity to targeted therapy.Variation of K1-K2 linkers led to AL776, our first optimized EGFR-c-Src targeting prototype.We now term this new targeting model consisting of designing a kinase inhibitor K1-K2 to target Kin-1 and Kin-2, and to further release two inhibitors K1 and K2 of the latter kinases, "type III combi-targeting".

View Article: PubMed Central - PubMed

Affiliation: Cancer Drug Research Laboratory, Department of Medicine, Division of Medical Oncology, McGill University Health Center/Royal Victoria Hospital, 687 Pine Avenue West Rm M7.19, Montreal, Quebec, H3A 1A1 Canada.

ABSTRACT
Cancer cells are characterized by a complex network of interrelated and compensatory signaling driven by multiple kinases that reduce their sensitivity to targeted therapy. Therefore, strategies directed at inhibiting two or more kinases are required to robustly block the growth of refractory tumour cells. Here we report on a novel strategy to promote sustained inhibition of two oncogenic kinases (Kin-1 and Kin-2) by designing a molecule K1-K2, termed "combi-molecule", to induce a tandem blockade of Kin-1 and Kin-2, as an intact structure and to be further hydrolyzed to two inhibitors K1 and K2 directed at Kin-1 and Kin-2, respectively. We chose to target EGFR (Kin-1) and c-Src (Kin-2), two tyrosine kinases known to synergize to promote tumour growth and progression. Variation of K1-K2 linkers led to AL776, our first optimized EGFR-c-Src targeting prototype. Here we showed that: (a) AL776 blocked EGFR and c-Src as an intact structure using an in vitro kinase assay (IC50 EGFR = 0.12 μM and IC50 c-Src = 3 nM), (b) it could release K1 (AL621, a nanomolar EGFR inhibitor) and K2 (dasatinib, a clinically approved Abl/c-Src inhibitor) by hydrolytic cleavage both in vitro and in vivo, (c) it could robustly inhibit phosphorylation of EGFR and c-Src (0.25-1 μM) in cells, (d) it induced 2-4 fold stronger growth inhibition than gefitinib or dasatinib and apoptosis at concentrations as low as 1 μM, and, (e) blocked motility and invasion at sub-micromolar doses in the highly invasive 4T1 and MDA-MB-231 cells. Despite its size (MW = 1032), AL776 blocked phosphorylation of EGFR and c-Src in 4T1 tumours in vivo. We now term this new targeting model consisting of designing a kinase inhibitor K1-K2 to target Kin-1 and Kin-2, and to further release two inhibitors K1 and K2 of the latter kinases, "type III combi-targeting".

No MeSH data available.


Related in: MedlinePlus