The conjugates therapeutic efficacy with targeting moieties was evaluated using prostate cancer cell lines with and without EDB-Fn overexpression (C4-2 and DU-145). Breast cancer cell lines with and without integrin v1 overexpression (MCF-7 and MDA-MB-231) were also used to evaluate antiproliferative activity. Noncancerous embryonic kidney epithelial cell line (HEK-293) was used as a control since this cell line did not express both biomarkers. All the compounds with and without targeting moieties showed comparable or lower antiproliferative activity than the CBT alone after 72 h incubation. The antiproliferative activity of compound 11 on MCF-7 (IC50 = 160.4 nM) was 1.6 times greater than on MDA-MB-231 (IC50 = 251.6 nM), showing a slight improvement in antiproliferative activity. A similar observation was found with compound 16 that showed 1.45 times higher antiproliferative activity on MCF-7 (IC50 = 162.4 nM) as compared to MDA-MB-231 cells (IC50 = 235.0 nM). Both MCF-7 and MDA-MB-231 are breast cancer cell lines. MDA-MB-231 is triple-negative breast cancer cells, and it showed higher IC50 values for CBT than MCF-7 cells. Later, the antiproliferative activity was compared between MCF-7 and the control cells (HEK-293); it was observed that compound 11 showed 3.33 times higher antiproliferative activity on MCF-7 cells (IC50 for 11 = 160.4 nM) compared to HEK-293 (IC50 for 11 = 534.5 nM), and compound 16 showed 4.6 times higher antiproliferative activity on MCF-7 (IC50 for 16 = 162.4 nM) compared to HEK-293 (IC50 for 16 = 483.3 nM). Thereafter, we wanted to assess the effectiveness of the EDB-Fn targeting moiety by comparing the antiproliferative activities on prostate cancer cell lines with and without EDB-Fn overexpression. The data showed a similar trend to that observed in breast cancer cells. For instance, compound 11 showed 3.4 times higher antiproliferative activity on C4-2 (IC50 for 11 = 181.4 nM) compared to DU-145 (IC50 for 11 = 611.2 nM). Similarly, compound 16 showed 3.7 times higher antiproliferative activity on C4-2 (IC50 for 16 = 202.2 nM) compared to DU-145 (IC50 for 16 = 751.1 nM). Compound 16 (possessing EDB-Fn targeting moiety) showed 1.1 times less antiproliferative activity compared to compound 6 (cCPP-CBT, without targeting moiety) on C4-2 cell lines, but compound 16 showed a much higher difference (3.6 times) in antiproliferative activity when compared with compound 6 on EDB-Fn moderately expressing DU-145 cell line. This data is further corroborated in EDB-Fn nonexpressing HEK-293. For instance, compound 16 showed 4.6 times less antiproliferative activity when compared with compound 6 on HEK-293 cell line. It can be inferred from the data (Table 2) that the EDB-Fn targeting conjugate was significantly effective on EDB-Fn overexpressing cells, while the conjugate has minimal effect on nonexpressing or moderately expressing cells. In conclusion, it is plausible to state that the presence of conjugating moieties on compound 11 (integrin targeting) and compound 16 (EDB-Fn targeting) significantly affects their ability to adhere and deliver the chemotherapeutic agent selectively to biomarker overexpressing cancer cells.

The conjugates therapeutic efficacy with targeting moieties was evaluated using prostate cancer cell lines with and without EDB-Fn overexpression (C4-2 and DU-145). Breast cancer cell lines with and without integrin v1 overexpression (MCF-7 and MDA-MB-231) were also used to evaluate antiproliferative activity. Noncancerous embryonic kidney epithelial cell line (HEK-293) was used as a control since this cell line did not express both biomarkers. All the compounds with and without targeting moieties showed comparable or lower antiproliferative activity than the CBT alone after 72 h incubation. The antiproliferative activity of compound 11 on MCF-7 (IC50 = 160.4 nM) was 1.6 times greater than on MDA-MB-231 (IC50 = 251.6 nM), showing a slight improvement in antiproliferative activity. A similar observation was found with compound 16 that showed 1.45 times higher antiproliferative activity on MCF-7 (IC50 = 162.4 nM) as compared to MDA-MB-231 cells (IC50 = 235.0 nM). Both MCF-7 and MDA-MB-231 are breast cancer cell lines. MDA-MB-231 is triple-negative breast cancer cells, and it showed higher IC50 values for CBT than MCF-7 cells. Later, the antiproliferative activity was compared between MCF-7 and the control cells (HEK-293); it was observed that compound 11 showed 3.33 times higher antiproliferative activity on MCF-7 cells (IC50 for 11 = 160.4 nM) compared to HEK-293 (IC50 for 11 = 534.5 nM), and compound 16 showed 4.6 times higher antiproliferative activity on MCF-7 (IC50 for 16 = 162.4 nM) compared to HEK-293 (IC50 for 16 = 483.3 nM). Thereafter, we wanted to assess the effectiveness of the EDB-Fn targeting moiety by comparing the antiproliferative activities on prostate cancer cell lines with and without EDB-Fn overexpression. The data showed a similar trend to that observed in breast cancer cells. For instance, compound 11 showed 3.4 times higher antiproliferative activity on C4-2 (IC50 for 11 = 181.4 nM) compared to DU-145 (IC50 for 11 = 611.2 nM). Similarly, compound 16 showed 3.7 times higher antiproliferative activity on C4-2 (IC50 for 16 = 202.2 nM) compared to DU-145 (IC50 for 16 = 751.1 nM). Compound 16 (possessing EDB-Fn targeting moiety) showed 1.1 times less antiproliferative activity compared to compound 6 (cCPP-CBT, without targeting moiety) on C4-2 cell lines, but compound 16 showed a much higher difference (3.6 times) in antiproliferative activity when compared with compound 6 on EDB-Fn moderately expressing DU-145 cell line. This data is further corroborated in EDB-Fn nonexpressing HEK-293. For instance, compound 16 showed 4.6 times less antiproliferative activity when compared with compound 6 on HEK-293 cell line. It can be inferred from the data (Table 2) that the EDB-Fn targeting conjugate was significantly effective on EDB-Fn overexpressing cells, while the conjugate has minimal effect on nonexpressing or moderately expressing cells. In conclusion, it is plausible to state that the presence of conjugating moieties on compound 11 (integrin targeting) and compound 16 (EDB-Fn targeting) significantly affects their ability to adhere and deliver the chemotherapeutic agent selectively to biomarker overexpressing cancer cells.

The conjugates therapeutic efficacy with targeting moieties was evaluated using prostate cancer cell lines with and without EDB-Fn overexpression (C4-2 and DU-145). Breast cancer cell lines with and without integrin v1 overexpression (MCF-7 and MDA-MB-231) were also used to evaluate antiproliferative activity. Noncancerous embryonic kidney epithelial cell line (HEK-293) was used as a control since this cell line did not express both biomarkers. All the compounds with and without targeting moieties showed comparable or lower antiproliferative activity than the CBT alone after 72 h incubation. The antiproliferative activity of compound 11 on MCF-7 (IC50 = 160.4 nM) was 1.6 times greater than on MDA-MB-231 (IC50 = 251.6 nM), showing a slight improvement in antiproliferative activity. A similar observation was found with compound 16 that showed 1.45 times higher antiproliferative activity on MCF-7 (IC50 = 162.4 nM) as compared to MDA-MB-231 cells (IC50 = 235.0 nM). Both MCF-7 and MDA-MB-231 are breast cancer cell lines. MDA-MB-231 is triple-negative breast cancer cells, and it showed higher IC50 values for CBT than MCF-7 cells. Later, the antiproliferative activity was compared between MCF-7 and the control cells (HEK-293); it was observed that compound 11 showed 3.33 times higher antiproliferative activity on MCF-7 cells (IC50 for 11 = 160.4 nM) compared to HEK-293 (IC50 for 11 = 534.5 nM), and compound 16 showed 4.6 times higher antiproliferative activity on MCF-7 (IC50 for 16 = 162.4 nM) compared to HEK-293 (IC50 for 16 = 483.3 nM). Thereafter, we wanted to assess the effectiveness of the EDB-Fn targeting moiety by comparing the antiproliferative activities on prostate cancer cell lines with and without EDB-Fn overexpression. The data showed a similar trend to that observed in breast cancer cells. For instance, compound 11 showed 3.4 times higher antiproliferative activity on C4-2 (IC50 for 11 = 181.4 nM) compared to DU-145 (IC50 for 11 = 611.2 nM). Similarly, compound 16 showed 3.7 times higher antiproliferative activity on C4-2 (IC50 for 16 = 202.2 nM) compared to DU-145 (IC50 for 16 = 751.1 nM). Compound 16 (possessing EDB-Fn targeting moiety) showed 1.1 times less antiproliferative activity compared to compound 6 (cCPP-CBT, without targeting moiety) on C4-2 cell lines, but compound 16 showed a much higher difference (3.6 times) in antiproliferative activity when compared with compound 6 on EDB-Fn moderately expressing DU-145 cell line. This data is further corroborated in EDB-Fn nonexpressing HEK-293. For instance, compound 16 showed 4.6 times less antiproliferative activity when compared with compound 6 on HEK-293 cell line. It can be inferred from the data (Table 2) that the EDB-Fn targeting conjugate was significantly effective on EDB-Fn overexpressing cells, while the conjugate has minimal effect on nonexpressing or moderately expressing cells. In conclusion, it is plausible to state that the presence of conjugating moieties on compound 11 (integrin targeting) and compound 16 (EDB-Fn targeting) significantly affects their ability to adhere and deliver the chemotherapeutic agent selectively to biomarker overexpressing cancer cells.

The conjugates therapeutic efficacy with targeting moieties was evaluated using prostate cancer cell lines with and without EDB-Fn overexpression (C4-2 and DU-145). Breast cancer cell lines with and without integrin v1 overexpression (MCF-7 and MDA-MB-231) were also used to evaluate antiproliferative activity. Noncancerous embryonic kidney epithelial cell line (HEK-293) was used as a control since this cell line did not express both biomarkers. All the compounds with and without targeting moieties showed comparable or lower antiproliferative activity than the CBT alone after 72 h incubation. The antiproliferative activity of compound 11 on MCF-7 (IC50 = 160.4 nM) was 1.6 times greater than on MDA-MB-231 (IC50 = 251.6 nM), showing a slight improvement in antiproliferative activity. A similar observation was found with compound 16 that showed 1.45 times higher antiproliferative activity on MCF-7 (IC50 = 162.4 nM) as compared to MDA-MB-231 cells (IC50 = 235.0 nM). Both MCF-7 and MDA-MB-231 are breast cancer cell lines. MDA-MB-231 is triple-negative breast cancer cells, and it showed higher IC50 values for CBT than MCF-7 cells. Later, the antiproliferative activity was compared between MCF-7 and the control cells (HEK-293); it was observed that compound 11 showed 3.33 times higher antiproliferative activity on MCF-7 cells (IC50 for 11 = 160.4 nM) compared to HEK-293 (IC50 for 11 = 534.5 nM), and compound 16 showed 4.6 times higher antiproliferative activity on MCF-7 (IC50 for 16 = 162.4 nM) compared to HEK-293 (IC50 for 16 = 483.3 nM). Thereafter, we wanted to assess the effectiveness of the EDB-Fn targeting moiety by comparing the antiproliferative activities on prostate cancer cell lines with and without EDB-Fn overexpression. The data showed a similar trend to that observed in breast cancer cells. For instance, compound 11 showed 3.4 times higher antiproliferative activity on C4-2 (IC50 for 11 = 181.4 nM) compared to DU-145 (IC50 for 11 = 611.2 nM). Similarly, compound 16 showed 3.7 times higher antiproliferative activity on C4-2 (IC50 for 16 = 202.2 nM) compared to DU-145 (IC50 for 16 = 751.1 nM). Compound 16 (possessing EDB-Fn targeting moiety) showed 1.1 times less antiproliferative activity compared to compound 6 (cCPP-CBT, without targeting moiety) on C4-2 cell lines, but compound 16 showed a much higher difference (3.6 times) in antiproliferative activity when compared with compound 6 on EDB-Fn moderately expressing DU-145 cell line. This data is further corroborated in EDB-Fn nonexpressing HEK-293. For instance, compound 16 showed 4.6 times less antiproliferative activity when compared with compound 6 on HEK-293 cell line. It can be inferred from the data (Table 2) that the EDB-Fn targeting conjugate was significantly effective on EDB-Fn overexpressing cells, while the conjugate has minimal effect on nonexpressing or moderately expressing cells. In conclusion, it is plausible to state that the presence of conjugating moieties on compound 11 (integrin targeting) and compound 16 (EDB-Fn targeting) significantly affects their ability to adhere and deliver the chemotherapeutic agent selectively to biomarker overexpressing cancer cells.

The conjugates therapeutic efficacy with targeting moieties was evaluated using prostate cancer cell lines with and without EDB-Fn overexpression (C4-2 and DU-145). Breast cancer cell lines with and without integrin v1 overexpression (MCF-7 and MDA-MB-231) were also used to evaluate antiproliferative activity. Noncancerous embryonic kidney epithelial cell line (HEK-293) was used as a control since this cell line did not express both biomarkers. All the compounds with and without targeting moieties showed comparable or lower antiproliferative activity than the CBT alone after 72 h incubation. The antiproliferative activity of compound 11 on MCF-7 (IC50 = 160.4 nM) was 1.6 times greater than on MDA-MB-231 (IC50 = 251.6 nM), showing a slight improvement in antiproliferative activity. A similar observation was found with compound 16 that showed 1.45 times higher antiproliferative activity on MCF-7 (IC50 = 162.4 nM) as compared to MDA-MB-231 cells (IC50 = 235.0 nM). Both MCF-7 and MDA-MB-231 are breast cancer cell lines. MDA-MB-231 is triple-negative breast cancer cells, and it showed higher IC50 values for CBT than MCF-7 cells. Later, the antiproliferative activity was compared between MCF-7 and the control cells (HEK-293); it was observed that compound 11 showed 3.33 times higher antiproliferative activity on MCF-7 cells (IC50 for 11 = 160.4 nM) compared to HEK-293 (IC50 for 11 = 534.5 nM), and compound 16 showed 4.6 times higher antiproliferative activity on MCF-7 (IC50 for 16 = 162.4 nM) compared to HEK-293 (IC50 for 16 = 483.3 nM). Thereafter, we wanted to assess the effectiveness of the EDB-Fn targeting moiety by comparing the antiproliferative activities on prostate cancer cell lines with and without EDB-Fn overexpression. The data showed a similar trend to that observed in breast cancer cells. For instance, compound 11 showed 3.4 times higher antiproliferative activity on C4-2 (IC50 for 11 = 181.4 nM) compared to DU-145 (IC50 for 11 = 611.2 nM). Similarly, compound 16 showed 3.7 times higher antiproliferative activity on C4-2 (IC50 for 16 = 202.2 nM) compared to DU-145 (IC50 for 16 = 751.1 nM). Compound 16 (possessing EDB-Fn targeting moiety) showed 1.1 times less antiproliferative activity compared to compound 6 (cCPP-CBT, without targeting moiety) on C4-2 cell lines, but compound 16 showed a much higher difference (3.6 times) in antiproliferative activity when compared with compound 6 on EDB-Fn moderately expressing DU-145 cell line. This data is further corroborated in EDB-Fn nonexpressing HEK-293. For instance, compound 16 showed 4.6 times less antiproliferative activity when compared with compound 6 on HEK-293 cell line. It can be inferred from the data (Table 2) that the EDB-Fn targeting conjugate was significantly effective on EDB-Fn overexpressing cells, while the conjugate has minimal effect on nonexpressing or moderately expressing cells. In conclusion, it is plausible to state that the presence of conjugating moieties on compound 11 (integrin targeting) and compound 16 (EDB-Fn targeting) significantly affects their ability to adhere and deliver the chemotherapeutic agent selectively to biomarker overexpressing cancer cells.