Linker ID	LIN00091
Linker Name	Disulfide bond
Linker Type	GSH concentration-sensitive linkers
Structure
	2D MOL
Formula	*2S2
#Ro5 Violations (Lipinski): 0	Molecular Weight (mw)			64.134
	Lipid-water partition coefficient (xlogp)			1.2964
	Hydrogen Bond Donor Count (hbonddonor)			0
	Hydrogen Bond Acceptor Count (hbondacc)			2
	Rotatable Bond Count (rotbonds)			1
Canonical smiles	*SS*

Experiment 1 Reporting the Activity Data of This PDC					[1]
Indication	Solid tumor
Efficacy Data	Grade 3 increased GGT	8.30%
Patients Enrolled	24 patients with various types of solid tumors.
Administration Dosage	9.6 mg/m2 BIW
MOA of PDC	BT1718 is a novel first in class bicyclic targeting peptide that selectively binds MT1 MMP (MMP-14) and is linked to the maytansinoid tubulin inhibitor DM1 by a cleavable disulfide linker. Bicycle Toxin Conjugates have a low molecular weight compared to other conjugated toxin approaches, enabling rapid tumour penetration and a short systemic half-life (up to 40 minutes in non-human primates) potentially reducing toxicity. The target MT1-MMP (MT1) is a surface metalloproteinase involved in tissue remodelling through proteolysis of extracellular matrix components: Highly expressed in tumours with unmet medical need, such as triple negative breast cancer, non small cell lung cancer and ovarian cancer. Strong link with cell invasion and metastasis. High tumour MT1 expression is correlated with poor outcomes in multiple tumour types. High adjacent stromal expression and low expression in adult normal tissue.    Click to Show/Hide
Description	24 patients were enrolled with various types of solid tumors across both dose escalation cohorts (see table). BIW RP2D was determined as 7.2 mg/m2. The 2 patients with DLTs at 9.6 mg/m2 BIW experienced grade 3 increased GGT or fatigue. QW dose escalation continues at 20 mg/m2. Mean number of cycles received = 2.3 months (N = 24), with no objective responses observed to date in this unselected population.    Click to Show/Hide
Experiment 2 Reporting the Activity Data of This PDC					[1]
Indication	Solid tumor
Efficacy Data	Fatigue	8.30%
Patients Enrolled	24 patients with various types of solid tumors.
Administration Dosage	9.6 mg/m2 BIW
MOA of PDC	BT1718 is a novel first in class bicyclic targeting peptide that selectively binds MT1 MMP (MMP-14) and is linked to the maytansinoid tubulin inhibitor DM1 by a cleavable disulfide linker. Bicycle Toxin Conjugates have a low molecular weight compared to other conjugated toxin approaches, enabling rapid tumour penetration and a short systemic half-life (up to 40 minutes in non-human primates) potentially reducing toxicity. The target MT1-MMP (MT1) is a surface metalloproteinase involved in tissue remodelling through proteolysis of extracellular matrix components: Highly expressed in tumours with unmet medical need, such as triple negative breast cancer, non small cell lung cancer and ovarian cancer. Strong link with cell invasion and metastasis. High tumour MT1 expression is correlated with poor outcomes in multiple tumour types. High adjacent stromal expression and low expression in adult normal tissue.    Click to Show/Hide
Description	24 patients were enrolled with various types of solid tumors across both dose escalation cohorts (see table). BIW RP2D was determined as 7.2 mg/m2. The 2 patients with DLTs at 9.6 mg/m2 BIW experienced grade 3 increased GGT or fatigue. QW dose escalation continues at 20 mg/m2. Mean number of cycles received = 2.3 months (N = 24), with no objective responses observed to date in this unselected population.    Click to Show/Hide

Experiment 1 Reporting the Activity Data of This PDC					[2]
Indication	Solid tumor
Efficacy Data	Tumor Growth Inhibition value (TGI)	12.50%
Administration Time	Twice a week for 12 days
Administration Dosage	1 mg/kg
Evaluation Method	Tumor volume detection
MOA of PDC	BT1718 is a novel first in class bicyclic targeting peptide that selectively binds MT1 MMP (MMP-14) and is linked to the maytansinoid tubulin inhibitor DM1 by a cleavable disulfide linker. Bicycle Toxin Conjugates have a low molecular weight compared to other conjugated toxin approaches, enabling rapid tumour penetration and a short systemic half-life (up to 40 minutes in non-human primates) potentially reducing toxicity. The target MT1-MMP (MT1) is a surface metalloproteinase involved in tissue remodelling through proteolysis of extracellular matrix components: Highly expressed in tumours with unmet medical need, such as triple negative breast cancer, non small cell lung cancer and ovarian cancer. Strong link with cell invasion and metastasis. High tumour MT1 expression is correlated with poor outcomes in multiple tumour types. High adjacent stromal expression and low expression in adult normal tissue.    Click to Show/Hide
Description	Testing of BT1718 in different dosing regimes in an additional model (HT-1080 cells) also demonstrated excellent tumour regression, with 10mg/kg biw leading to complete tumour clearance in all 3 animals within 23 days and no re-growth out to 72 days.
In Vivo Model	Mice implanted with MT1-positive HT-1080 cells
In Vitro Model	Fibrosarcoma	HT-1080 cell		CVCL_0317
Experiment 2 Reporting the Activity Data of This PDC					[2]
Indication	Solid tumor
Efficacy Data	Tumor Growth Inhibition value (TGI)	12.70%
Administration Time	Three times a week for 12 days
Administration Dosage	1 mg/kg
Evaluation Method	Tumor volume detection
MOA of PDC	BT1718 is a novel first in class bicyclic targeting peptide that selectively binds MT1 MMP (MMP-14) and is linked to the maytansinoid tubulin inhibitor DM1 by a cleavable disulfide linker. Bicycle Toxin Conjugates have a low molecular weight compared to other conjugated toxin approaches, enabling rapid tumour penetration and a short systemic half-life (up to 40 minutes in non-human primates) potentially reducing toxicity. The target MT1-MMP (MT1) is a surface metalloproteinase involved in tissue remodelling through proteolysis of extracellular matrix components: Highly expressed in tumours with unmet medical need, such as triple negative breast cancer, non small cell lung cancer and ovarian cancer. Strong link with cell invasion and metastasis. High tumour MT1 expression is correlated with poor outcomes in multiple tumour types. High adjacent stromal expression and low expression in adult normal tissue.    Click to Show/Hide
Description	Testing of BT1718 in different dosing regimes in an additional model (HT-1080 cells) also demonstrated excellent tumour regression, with 10mg/kg biw leading to complete tumour clearance in all 3 animals within 23 days and no re-growth out to 75 days.
In Vivo Model	Mice implanted with MT1-positive HT-1080 cells
In Vitro Model	Fibrosarcoma	HT-1080 cell		CVCL_0317
Experiment 3 Reporting the Activity Data of This PDC					[1]
Indication	Fibrosarcoma
Efficacy Data	Tumor Growth Inhibition value (TGI)	18%
Administration Time	14 days
Administration Dosage	1 mg/kg qw
MOA of PDC	BT1718 is a novel first in class bicyclic targeting peptide that selectively binds MT1 MMP (MMP-14) and is linked to the maytansinoid tubulin inhibitor DM1 by a cleavable disulfide linker. Bicycle Toxin Conjugates have a low molecular weight compared to other conjugated toxin approaches, enabling rapid tumour penetration and a short systemic half-life (up to 40 minutes in non-human primates) potentially reducing toxicity. The target MT1-MMP (MT1) is a surface metalloproteinase involved in tissue remodelling through proteolysis of extracellular matrix components: Highly expressed in tumours with unmet medical need, such as triple negative breast cancer, non small cell lung cancer and ovarian cancer. Strong link with cell invasion and metastasis. High tumour MT1 expression is correlated with poor outcomes in multiple tumour types. High adjacent stromal expression and low expression in adult normal tissue.    Click to Show/Hide
Description	Treatment with BDCs containing the most labile linkers (BT17BDC17 or BT1718) showed rapid and complete tumour clearance (EBC-1 cells), while BDCs containing more stabilised linkers showed comparatively reduced efficacy (fig. 5) suggesting that target internalisation is not the sole mechanism of action for BDC efficacy and that extracellular cleavage and release of toxin within the local tumour environment likely also contribute. Only the most labile BDC (BT17BDC17) caused any significant toxicity (17% 9.7 body weight loss); all others were well tolerated (<10% body weight loss at 10 mg/kg tiw). Optimal therapeutic index was achieved with BT1718. Testing of BT1718 in different dosing regimes in an additional model (HT-1080 cells) also demonstrated excellent tumour regression, with 10 mg/kg biw leading to complete tumour clearance in all 3 animals within 23 days and no re-growth out to 70 days.    Click to Show/Hide
In Vivo Model	Cell-derived xenograftmodel inmiceimplanted with MT1-positive HT-1080 cells.
In Vitro Model	Fibrosarcoma	HT-1080 cell		CVCL_0317
Experiment 4 Reporting the Activity Data of This PDC					[1]
Indication	Fibrosarcoma
Efficacy Data	Tumor Growth Inhibition value (TGI)	27%
Administration Time	14 days
Administration Dosage	1 mg/kg biw
MOA of PDC	BT1718 is a novel first in class bicyclic targeting peptide that selectively binds MT1 MMP (MMP-14) and is linked to the maytansinoid tubulin inhibitor DM1 by a cleavable disulfide linker. Bicycle Toxin Conjugates have a low molecular weight compared to other conjugated toxin approaches, enabling rapid tumour penetration and a short systemic half-life (up to 40 minutes in non-human primates) potentially reducing toxicity. The target MT1-MMP (MT1) is a surface metalloproteinase involved in tissue remodelling through proteolysis of extracellular matrix components: Highly expressed in tumours with unmet medical need, such as triple negative breast cancer, non small cell lung cancer and ovarian cancer. Strong link with cell invasion and metastasis. High tumour MT1 expression is correlated with poor outcomes in multiple tumour types. High adjacent stromal expression and low expression in adult normal tissue.    Click to Show/Hide
Description	Treatment with BDCs containing the most labile linkers (BT17BDC17 or BT1718) showed rapid and complete tumour clearance (EBC-1 cells), while BDCs containing more stabilised linkers showed comparatively reduced efficacy (fig. 5) suggesting that target internalisation is not the sole mechanism of action for BDC efficacy and that extracellular cleavage and release of toxin within the local tumour environment likely also contribute. Only the most labile BDC (BT17BDC17) caused any significant toxicity (17% 9.7 body weight loss); all others were well tolerated (<10% body weight loss at 10 mg/kg tiw). Optimal therapeutic index was achieved with BT1718. Testing of BT1718 in different dosing regimes in an additional model (HT-1080 cells) also demonstrated excellent tumour regression, with 10 mg/kg biw leading to complete tumour clearance in all 3 animals within 23 days and no re-growth out to 70 days.    Click to Show/Hide
In Vivo Model	Cell-derived xenograftmodel inmiceimplanted with MT1-positive HT-1080 cells.
In Vitro Model	Fibrosarcoma	HT-1080 cell		CVCL_0317
Experiment 5 Reporting the Activity Data of This PDC					[2]
Indication	Solid tumor
Efficacy Data	Tumor Growth Inhibition value (TGI)	85.40%
Administration Time	Three times a week for 12 days
Administration Dosage	3 mg/kg
Evaluation Method	Tumor volume detection
MOA of PDC	BT1718 is a novel first in class bicyclic targeting peptide that selectively binds MT1 MMP (MMP-14) and is linked to the maytansinoid tubulin inhibitor DM1 by a cleavable disulfide linker. Bicycle Toxin Conjugates have a low molecular weight compared to other conjugated toxin approaches, enabling rapid tumour penetration and a short systemic half-life (up to 40 minutes in non-human primates) potentially reducing toxicity. The target MT1-MMP (MT1) is a surface metalloproteinase involved in tissue remodelling through proteolysis of extracellular matrix components: Highly expressed in tumours with unmet medical need, such as triple negative breast cancer, non small cell lung cancer and ovarian cancer. Strong link with cell invasion and metastasis. High tumour MT1 expression is correlated with poor outcomes in multiple tumour types. High adjacent stromal expression and low expression in adult normal tissue.    Click to Show/Hide
Description	Testing of BT1718 in different dosing regimes in an additional model (HT-1080 cells) also demonstrated excellent tumour regression, with 10mg/kg biw leading to complete tumour clearance in all 3 animals within 23 days and no re-growth out to 74 days.
In Vivo Model	Mice implanted with MT1-positive HT-1080 cells
In Vitro Model	Fibrosarcoma	HT-1080 cell		CVCL_0317
Experiment 6 Reporting the Activity Data of This PDC					[1]
Indication	Fibrosarcoma
Efficacy Data	Tumor Growth Inhibition value (TGI)	88%
Administration Time	14 days
Administration Dosage	3 mg/kg qw
MOA of PDC	BT1718 is a novel first in class bicyclic targeting peptide that selectively binds MT1 MMP (MMP-14) and is linked to the maytansinoid tubulin inhibitor DM1 by a cleavable disulfide linker. Bicycle Toxin Conjugates have a low molecular weight compared to other conjugated toxin approaches, enabling rapid tumour penetration and a short systemic half-life (up to 40 minutes in non-human primates) potentially reducing toxicity. The target MT1-MMP (MT1) is a surface metalloproteinase involved in tissue remodelling through proteolysis of extracellular matrix components: Highly expressed in tumours with unmet medical need, such as triple negative breast cancer, non small cell lung cancer and ovarian cancer. Strong link with cell invasion and metastasis. High tumour MT1 expression is correlated with poor outcomes in multiple tumour types. High adjacent stromal expression and low expression in adult normal tissue.    Click to Show/Hide
Description	Treatment with BDCs containing the most labile linkers (BT17BDC17 or BT1718) showed rapid and complete tumour clearance (EBC-1 cells), while BDCs containing more stabilised linkers showed comparatively reduced efficacy (fig. 5) suggesting that target internalisation is not the sole mechanism of action for BDC efficacy and that extracellular cleavage and release of toxin within the local tumour environment likely also contribute. Only the most labile BDC (BT17BDC17) caused any significant toxicity (17% 9.7 body weight loss); all others were well tolerated (<10% body weight loss at 10 mg/kg tiw). Optimal therapeutic index was achieved with BT1718. Testing of BT1718 in different dosing regimes in an additional model (HT-1080 cells) also demonstrated excellent tumour regression, with 10 mg/kg biw leading to complete tumour clearance in all 3 animals within 23 days and no re-growth out to 70 days.    Click to Show/Hide
In Vivo Model	Cell-derived xenograftmodel inmiceimplanted with MT1-positive HT-1080 cells.
In Vitro Model	Fibrosarcoma	HT-1080 cell		CVCL_0317
Experiment 7 Reporting the Activity Data of This PDC					[1]
Indication	Lung squamous cell carcinoma
Efficacy Data	Tumor Growth Inhibition value (TGI)	100%
Administration Time	14 days
Administration Dosage	10mg/kg
MOA of PDC	BT1718 is a novel first in class bicyclic targeting peptide that selectively binds MT1 MMP (MMP-14) and is linked to the maytansinoid tubulin inhibitor DM1 by a cleavable disulfide linker. Bicycle Toxin Conjugates have a low molecular weight compared to other conjugated toxin approaches, enabling rapid tumour penetration and a short systemic half-life (up to 40 minutes in non-human primates) potentially reducing toxicity. The target MT1-MMP (MT1) is a surface metalloproteinase involved in tissue remodelling through proteolysis of extracellular matrix components: Highly expressed in tumours with unmet medical need, such as triple negative breast cancer, non small cell lung cancer and ovarian cancer. Strong link with cell invasion and metastasis. High tumour MT1 expression is correlated with poor outcomes in multiple tumour types. High adjacent stromal expression and low expression in adult normal tissue.    Click to Show/Hide
Description	Treatment with BDCs containing the most labile linkers (BT17BDC17 or BT1718) showed rapid and complete tumour clearance (EBC-1 cells), while BDCs containing more stabilised linkers showed comparatively reduced efficacy (fig. 5) suggesting that target internalisation is not the sole mechanism of action for BDC efficacy and that extracellular cleavage and release of toxin within the local tumour environment likely also contribute. Only the most labile BDC (BT17BDC17) caused any significant toxicity (17% 9.7 body weight loss); all others were well tolerated (<10% body weight loss at 10 mg/kg tiw). Optimal therapeutic index was achieved with BT1718. Testing of BT1718 in different dosing regimes in an additional model (HT-1080 cells) also demonstrated excellent tumour regression, with 10 mg/kg biw leading to complete tumour clearance in all 3 animals within 23 days and no re-growth out to 70 days.    Click to Show/Hide
In Vivo Model	Cell-derived xenograftmodel inmiceimplanted with MT1-positive EBC-1 cells.
In Vitro Model	Lung squamous cell carcinoma	EBC-1 cell		CVCL_2891
Experiment 8 Reporting the Activity Data of This PDC					[1]
Indication	Fibrosarcoma
Efficacy Data	Tumor Growth Inhibition value (TGI)	100%
Administration Time	14 days
Administration Dosage	10 mg/kg biw
MOA of PDC	BT1718 is a novel first in class bicyclic targeting peptide that selectively binds MT1 MMP (MMP-14) and is linked to the maytansinoid tubulin inhibitor DM1 by a cleavable disulfide linker. Bicycle Toxin Conjugates have a low molecular weight compared to other conjugated toxin approaches, enabling rapid tumour penetration and a short systemic half-life (up to 40 minutes in non-human primates) potentially reducing toxicity. The target MT1-MMP (MT1) is a surface metalloproteinase involved in tissue remodelling through proteolysis of extracellular matrix components: Highly expressed in tumours with unmet medical need, such as triple negative breast cancer, non small cell lung cancer and ovarian cancer. Strong link with cell invasion and metastasis. High tumour MT1 expression is correlated with poor outcomes in multiple tumour types. High adjacent stromal expression and low expression in adult normal tissue.    Click to Show/Hide
Description	Treatment with BDCs containing the most labile linkers (BT17BDC17 or BT1718) showed rapid and complete tumour clearance (EBC-1 cells), while BDCs containing more stabilised linkers showed comparatively reduced efficacy (fig. 5) suggesting that target internalisation is not the sole mechanism of action for BDC efficacy and that extracellular cleavage and release of toxin within the local tumour environment likely also contribute. Only the most labile BDC (BT17BDC17) caused any significant toxicity (17% 9.7 body weight loss); all others were well tolerated (<10% body weight loss at 10 mg/kg tiw). Optimal therapeutic index was achieved with BT1718. Testing of BT1718 in different dosing regimes in an additional model (HT-1080 cells) also demonstrated excellent tumour regression, with 10 mg/kg biw leading to complete tumour clearance in all 3 animals within 23 days and no re-growth out to 70 days.    Click to Show/Hide
In Vivo Model	Cell-derived xenograftmodel inmiceimplanted with MT1-positive HT-1080 cells.
In Vitro Model	Fibrosarcoma	HT-1080 cell		CVCL_0317
Experiment 9 Reporting the Activity Data of This PDC					[1]
Indication	Fibrosarcoma
Efficacy Data	Tumor Growth Inhibition value (TGI)	100%
Administration Time	14 days
Administration Dosage	3 mg/kg biw
MOA of PDC	BT1718 is a novel first in class bicyclic targeting peptide that selectively binds MT1 MMP (MMP-14) and is linked to the maytansinoid tubulin inhibitor DM1 by a cleavable disulfide linker. Bicycle Toxin Conjugates have a low molecular weight compared to other conjugated toxin approaches, enabling rapid tumour penetration and a short systemic half-life (up to 40 minutes in non-human primates) potentially reducing toxicity. The target MT1-MMP (MT1) is a surface metalloproteinase involved in tissue remodelling through proteolysis of extracellular matrix components: Highly expressed in tumours with unmet medical need, such as triple negative breast cancer, non small cell lung cancer and ovarian cancer. Strong link with cell invasion and metastasis. High tumour MT1 expression is correlated with poor outcomes in multiple tumour types. High adjacent stromal expression and low expression in adult normal tissue.    Click to Show/Hide
Description	Treatment with BDCs containing the most labile linkers (BT17BDC17 or BT1718) showed rapid and complete tumour clearance (EBC-1 cells), while BDCs containing more stabilised linkers showed comparatively reduced efficacy (fig. 5) suggesting that target internalisation is not the sole mechanism of action for BDC efficacy and that extracellular cleavage and release of toxin within the local tumour environment likely also contribute. Only the most labile BDC (BT17BDC17) caused any significant toxicity (17% 9.7 body weight loss); all others were well tolerated (<10% body weight loss at 10 mg/kg tiw). Optimal therapeutic index was achieved with BT1718. Testing of BT1718 in different dosing regimes in an additional model (HT-1080 cells) also demonstrated excellent tumour regression, with 10 mg/kg biw leading to complete tumour clearance in all 3 animals within 23 days and no re-growth out to 70 days.    Click to Show/Hide
In Vivo Model	Cell-derived xenograftmodel inmiceimplanted with MT1-positive HT-1080 cells.
In Vitro Model	Fibrosarcoma	HT-1080 cell		CVCL_0317
Experiment 10 Reporting the Activity Data of This PDC					[1]
Indication	Fibrosarcoma
Efficacy Data	Tumor Growth Inhibition value (TGI)	100%
Administration Time	14 days
Administration Dosage	10 mg/kg qw
MOA of PDC	BT1718 is a novel first in class bicyclic targeting peptide that selectively binds MT1 MMP (MMP-14) and is linked to the maytansinoid tubulin inhibitor DM1 by a cleavable disulfide linker. Bicycle Toxin Conjugates have a low molecular weight compared to other conjugated toxin approaches, enabling rapid tumour penetration and a short systemic half-life (up to 40 minutes in non-human primates) potentially reducing toxicity. The target MT1-MMP (MT1) is a surface metalloproteinase involved in tissue remodelling through proteolysis of extracellular matrix components: Highly expressed in tumours with unmet medical need, such as triple negative breast cancer, non small cell lung cancer and ovarian cancer. Strong link with cell invasion and metastasis. High tumour MT1 expression is correlated with poor outcomes in multiple tumour types. High adjacent stromal expression and low expression in adult normal tissue.    Click to Show/Hide
Description	Treatment with BDCs containing the most labile linkers (BT17BDC17 or BT1718) showed rapid and complete tumour clearance (EBC-1 cells), while BDCs containing more stabilised linkers showed comparatively reduced efficacy (fig. 5) suggesting that target internalisation is not the sole mechanism of action for BDC efficacy and that extracellular cleavage and release of toxin within the local tumour environment likely also contribute. Only the most labile BDC (BT17BDC17) caused any significant toxicity (17% 9.7 body weight loss); all others were well tolerated (<10% body weight loss at 10 mg/kg tiw). Optimal therapeutic index was achieved with BT1718. Testing of BT1718 in different dosing regimes in an additional model (HT-1080 cells) also demonstrated excellent tumour regression, with 10 mg/kg biw leading to complete tumour clearance in all 3 animals within 23 days and no re-growth out to 70 days.    Click to Show/Hide
In Vivo Model	Cell-derived xenograftmodel inmiceimplanted with MT1-positive HT-1080 cells.
In Vitro Model	Fibrosarcoma	HT-1080 cell		CVCL_0317
Experiment 11 Reporting the Activity Data of This PDC					[2]
Indication	Solid tumor
Efficacy Data	Tumor Growth Inhibition value (TGI)	100%
Administration Time	12 days
Administration Dosage	10 mg/kg
Evaluation Method	Tumor volume detection
MOA of PDC	BT1718 is a novel first in class bicyclic targeting peptide that selectively binds MT1 MMP (MMP-14) and is linked to the maytansinoid tubulin inhibitor DM1 by a cleavable disulfide linker. Bicycle Toxin Conjugates have a low molecular weight compared to other conjugated toxin approaches, enabling rapid tumour penetration and a short systemic half-life (up to 40 minutes in non-human primates) potentially reducing toxicity. The target MT1-MMP (MT1) is a surface metalloproteinase involved in tissue remodelling through proteolysis of extracellular matrix components: Highly expressed in tumours with unmet medical need, such as triple negative breast cancer, non small cell lung cancer and ovarian cancer. Strong link with cell invasion and metastasis. High tumour MT1 expression is correlated with poor outcomes in multiple tumour types. High adjacent stromal expression and low expression in adult normal tissue.    Click to Show/Hide
Description	Treatment with BDCs containing the most labile linkers (BT17BDC17 orBT1718) showed rapid and complete tumour clearance (EBC-1 cells).
In Vivo Model	Mice implanted with MT1-positive EBC-1 cells
In Vitro Model	Lung squamous cell carcinoma	EBC-1 cell		CVCL_2891
Experiment 12 Reporting the Activity Data of This PDC					[2]
Indication	Solid tumor
Efficacy Data	Tumor Growth Inhibition value (TGI)	100%
Administration Time	Twice a week for 12 days
Administration Dosage	10 mg/kg
Evaluation Method	Tumor volume detection
MOA of PDC	BT1718 is a novel first in class bicyclic targeting peptide that selectively binds MT1 MMP (MMP-14) and is linked to the maytansinoid tubulin inhibitor DM1 by a cleavable disulfide linker. Bicycle Toxin Conjugates have a low molecular weight compared to other conjugated toxin approaches, enabling rapid tumour penetration and a short systemic half-life (up to 40 minutes in non-human primates) potentially reducing toxicity. The target MT1-MMP (MT1) is a surface metalloproteinase involved in tissue remodelling through proteolysis of extracellular matrix components: Highly expressed in tumours with unmet medical need, such as triple negative breast cancer, non small cell lung cancer and ovarian cancer. Strong link with cell invasion and metastasis. High tumour MT1 expression is correlated with poor outcomes in multiple tumour types. High adjacent stromal expression and low expression in adult normal tissue.    Click to Show/Hide
Description	Testing of BT1718 in different dosing regimes in an additional model (HT-1080 cells) also demonstrated excellent tumour regression, with 10mg/kg biw leading to complete tumour clearance in all 3 animals within 23 days and no re-growth out to 70 days.
In Vivo Model	Mice implanted with MT1-positive HT-1080 cells
In Vitro Model	Fibrosarcoma	HT-1080 cell		CVCL_0317
Experiment 13 Reporting the Activity Data of This PDC					[2]
Indication	Solid tumor
Efficacy Data	Tumor Growth Inhibition value (TGI)	100%
Administration Time	Twice a week for 12 days
Administration Dosage	3 mg/kg
Evaluation Method	Tumor volume detection
MOA of PDC	BT1718 is a novel first in class bicyclic targeting peptide that selectively binds MT1 MMP (MMP-14) and is linked to the maytansinoid tubulin inhibitor DM1 by a cleavable disulfide linker. Bicycle Toxin Conjugates have a low molecular weight compared to other conjugated toxin approaches, enabling rapid tumour penetration and a short systemic half-life (up to 40 minutes in non-human primates) potentially reducing toxicity. The target MT1-MMP (MT1) is a surface metalloproteinase involved in tissue remodelling through proteolysis of extracellular matrix components: Highly expressed in tumours with unmet medical need, such as triple negative breast cancer, non small cell lung cancer and ovarian cancer. Strong link with cell invasion and metastasis. High tumour MT1 expression is correlated with poor outcomes in multiple tumour types. High adjacent stromal expression and low expression in adult normal tissue.    Click to Show/Hide
Description	Testing of BT1718 in different dosing regimes in an additional model (HT-1080 cells) also demonstrated excellent tumour regression, with 10mg/kg biw leading to complete tumour clearance in all 3 animals within 23 days and no re-growth out to 71 days.
In Vivo Model	Mice implanted with MT1-positive HT-1080 cells
In Vitro Model	Fibrosarcoma	HT-1080 cell		CVCL_0317
Experiment 14 Reporting the Activity Data of This PDC					[2]
Indication	Solid tumor
Efficacy Data	Tumor Growth Inhibition value (TGI)	100%
Administration Time	Three times a week for 12 days
Administration Dosage	10 mg/kg
Evaluation Method	Tumor volume detection
MOA of PDC	BT1718 is a novel first in class bicyclic targeting peptide that selectively binds MT1 MMP (MMP-14) and is linked to the maytansinoid tubulin inhibitor DM1 by a cleavable disulfide linker. Bicycle Toxin Conjugates have a low molecular weight compared to other conjugated toxin approaches, enabling rapid tumour penetration and a short systemic half-life (up to 40 minutes in non-human primates) potentially reducing toxicity. The target MT1-MMP (MT1) is a surface metalloproteinase involved in tissue remodelling through proteolysis of extracellular matrix components: Highly expressed in tumours with unmet medical need, such as triple negative breast cancer, non small cell lung cancer and ovarian cancer. Strong link with cell invasion and metastasis. High tumour MT1 expression is correlated with poor outcomes in multiple tumour types. High adjacent stromal expression and low expression in adult normal tissue.    Click to Show/Hide
Description	Testing of BT1718 in different dosing regimes in an additional model (HT-1080 cells) also demonstrated excellent tumour regression, with 10mg/kg biw leading to complete tumour clearance in all 3 animals within 23 days and no re-growth out to 73 days.
In Vivo Model	Mice implanted with MT1-positive HT-1080 cells
In Vitro Model	Fibrosarcoma	HT-1080 cell		CVCL_0317

Experiment 1 Reporting the Activity Data of This PDC					[3]
Indication	Tumor
Efficacy Data	Tumor Growth Inhibition value (TGI)	58.10%
Administration Time	23 days
Administration Dosage	5 mg/kg
Evaluation Method	Tumor volume detection
Description	Combination treatment significantly inhibited tumor growth without significant toxicity in both mouse xenografts compared with CBX-12 and ceralasertib monotherapy without significant toxicity.
In Vivo Model	HCT-116 xenografts
Experiment 2 Reporting the Activity Data of This PDC					[3]
Indication	Tumor
Efficacy Data	Tumor Growth Inhibition value (TGI)	76.50%
Administration Time	35 days
Administration Dosage	10 mg/kg
Evaluation Method	Tumor volume detection
Description	Combination treatment significantly inhibited tumor growth without significant toxicity in both mouse xenografts compared with CBX-12 and ceralasertib monotherapy without significant toxicity.
In Vivo Model	MDA-MB-231 xenografts
Experiment 3 Reporting the Activity Data of This PDC					[3]
Indication	Tumor
Efficacy Data	Tumor Growth Inhibition value (TGI)	81.40%
Administration Time	23 days
Administration Dosage	5 mg/kg with ceralasertib 25 mg/kg
Evaluation Method	Tumor volume detection
Description	Combination treatment significantly inhibited tumor growth without significant toxicity in both mouse xenografts compared with CBX-12 and ceralasertib monotherapy without significant toxicity.
In Vivo Model	HCT-116 xenografts
Experiment 4 Reporting the Activity Data of This PDC					[3]
Indication	Tumor
Efficacy Data	Tumor Growth Inhibition value (TGI)	95%
Administration Time	35 days
Administration Dosage	10 mg/kg with ceralasertib 25 mg/kg
Evaluation Method	Tumor volume detection
Description	Combination treatment significantly inhibited tumor growth without significant toxicity in both mouse xenografts compared with CBX-12 and ceralasertib monotherapy without significant toxicity.
In Vivo Model	MDA-MB-231 xenografts
Experiment 5 Reporting the Activity Data of This PDC					[3]
Indication	Tumor
Efficacy Data	Survival rate	60 mm3
Administration Time	45 days
Administration Dosage	5 mg/kg
Description	Combination treatment significantly inhibited tumor growth without significant toxicity in both mouse xenografts compared with CBX-12 and ceralasertib monotherapy without significant toxicity.
In Vivo Model	HCT-116 xenografts
Experiment 6 Reporting the Activity Data of This PDC					[3]
Indication	Tumor
Efficacy Data	Survival rate	75 mm3
Administration Time	42 days
Administration Dosage	10 mg/kg
Description	Combination treatment significantly inhibited tumor growth without significant toxicity in both mouse xenografts compared with CBX-12 and ceralasertib monotherapy without significant toxicity.
In Vivo Model	MDA-MB-231 xenografts
Experiment 7 Reporting the Activity Data of This PDC					[3]
Indication	Tumor
Efficacy Data	Survival rate	100 mm3
Administration Time	42 days
Administration Dosage	10 mg/kg with ceralasertib 25 mg/kg
Description	Combination treatment significantly inhibited tumor growth without significant toxicity in both mouse xenografts compared with CBX-12 and ceralasertib monotherapy without significant toxicity.
In Vivo Model	MDA-MB-231 xenografts
Experiment 8 Reporting the Activity Data of This PDC					[3]
Indication	Tumor
Efficacy Data	Survival rate	100 mm3
Administration Time	45 days
Administration Dosage	5 mg/kg with ceralasertib 25 mg/kg
Description	Combination treatment significantly inhibited tumor growth without significant toxicity in both mouse xenografts compared with CBX-12 and ceralasertib monotherapy without significant toxicity.
In Vivo Model	HCT-116 xenografts

Experiment 1 Reporting the Activity Data of This PDC					[4]
Indication	Small cell lung cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	0.06 µmol/L
Administration Time	6 h
Description	PEN-221 receptor dependent activity in vitro varied from strong (NCI-H524) to modest (HCC-33) but was consistent (N = 3) and correlated with in vivo activity (described below). Octreotide alone did not have an effect on cellular growth. These cell lines also demonstrated sensitivity to the cytotoxic payload, DM1, with sub-micromolar IC50's after a 6-hour exposure and 70-hour additional incubation period (Supplementary Fig. S2B).    Click to Show/Hide
In Vitro Model	Lung small cell carcinoma	NCI-H524 cell		CVCL_1568
Experiment 2 Reporting the Activity Data of This PDC					[4]
Indication	Small cell lung cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	0.108 µmol/L
Administration Time	6 h
Description	PEN-221 receptor dependent activity in vitro varied from strong (NCI-H524) to modest (HCC-33) but was consistent (N = 3) and correlated with in vivo activity (described below). Octreotide alone did not have an effect on cellular growth. These cell lines also demonstrated sensitivity to the cytotoxic payload, DM1, with sub-micromolar IC50's after a 6-hour exposure and 70-hour additional incubation period (Supplementary Fig. S2B).    Click to Show/Hide
In Vitro Model	Small cell lung carcinoma	NCI-H69 cell		CVCL_1579
Experiment 3 Reporting the Activity Data of This PDC					[4]
Indication	Small cell lung cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	0.258 µmol/L
Administration Time	6 h
Description	PEN-221 receptor dependent activity in vitro varied from strong (NCI-H524) to modest (HCC-33) but was consistent (N = 3) and correlated with in vivo activity (described below). Octreotide alone did not have an effect on cellular growth. These cell lines also demonstrated sensitivity to the cytotoxic payload, DM1, with sub-micromolar IC50's after a 6-hour exposure and 70-hour additional incubation period (Supplementary Fig. S2B).    Click to Show/Hide
In Vitro Model	Small cell lung carcinoma	NCI-H69 cell		CVCL_1579
Experiment 4 Reporting the Activity Data of This PDC					[4]
Indication	Small cell lung cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	0.333 µmol/L
Administration Time	6 h
Description	PEN-221 receptor dependent activity in vitro varied from strong (NCI-H524) to modest (HCC-33) but was consistent (N = 3) and correlated with in vivo activity (described below). Octreotide alone did not have an effect on cellular growth. These cell lines also demonstrated sensitivity to the cytotoxic payload, DM1, with sub-micromolar IC50's after a 6-hour exposure and 70-hour additional incubation period (Supplementary Fig. S2B).    Click to Show/Hide
In Vitro Model	Lung small cell carcinoma	NCI-H524 cell		CVCL_1568
Experiment 5 Reporting the Activity Data of This PDC					[4]
Indication	Small cell lung cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	0.658 µmol/L
Administration Time	6 h
Description	PEN-221 receptor dependent activity in vitro varied from strong (NCI-H524) to modest (HCC-33) but was consistent (N = 3) and correlated with in vivo activity (described below). Octreotide alone did not have an effect on cellular growth. These cell lines also demonstrated sensitivity to the cytotoxic payload, DM1, with sub-micromolar IC50's after a 6-hour exposure and 70-hour additional incubation period (Supplementary Fig. S2B).    Click to Show/Hide
In Vitro Model	Lung small cell carcinoma	HCC33 cell		CVCL_2058
Experiment 6 Reporting the Activity Data of This PDC					[4]
Indication	Small cell lung cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	0.83 µmol/L
Administration Time	6 h
Description	PEN-221 receptor dependent activity in vitro varied from strong (NCI-H524) to modest (HCC-33) but was consistent (N = 3) and correlated with in vivo activity (described below). Octreotide alone did not have an effect on cellular growth. These cell lines also demonstrated sensitivity to the cytotoxic payload, DM1, with sub-micromolar IC50's after a 6-hour exposure and 70-hour additional incubation period (Supplementary Fig. S2B).    Click to Show/Hide
In Vitro Model	Lung small cell carcinoma	HCC33 cell		CVCL_2058

Experiment 1 Reporting the Activity Data of This PDC					[5]
Indication	Obesity
Efficacy Data	Vehicle-corrected weight loss rate	9.50%
Administration Time	1 dose
Administration Dosage	0.22 nmol
Evaluation Method	This study revealed a superior vehicle-corrected weight loss of 9.5% in response to the GLP-1-MK-801 infusion relative to a weight loss of 4.5% after semaglutide infusion (Fig. 4b).
MOA of PDC	Treatment with GLP-1-inactive MK-801 produced no additional weight loss efficacy relative to GLP-1 monotherapy (Fig. 2g-i), indicating that the pronounced weight loss induced by GLP-1-MK-801 is driven by concerted and site-directed pharmacological GLP-1 receptor agonism and NMDA receptor antagonism.
In Vivo Model	High-fat high-sucrose-fed mice
Half life period	1.9 h
Experiment 2 Reporting the Activity Data of This PDC					[5]
Indication	Obesity
Efficacy Data	Vehicle-corrected weight loss rate	15.20%
Administration Time	Once a day for 9 days
Administration Dosage	100 nmol kg-1
Evaluation Method	We observed a pronounced vehicle-corrected weight loss of 15.2% in the group treated with GLP-1-MK-801 compared with 3.5% in the group treated with the parent GLP-1 analogue after 9days of treatment, underscoring that the weight-lowering efficacy of the conjugate is intact in the absence of functional MC4R signalling (Fig. 3l,m).
MOA of PDC	Treatment with GLP-1-inactive MK-801 produced no additional weight loss efficacy relative to GLP-1 monotherapy (Fig. 2g-i), indicating that the pronounced weight loss induced by GLP-1-MK-801 is driven by concerted and site-directed pharmacological GLP-1 receptor agonism and NMDA receptor antagonism.
In Vivo Model	Mc4r-KO mice
Half life period	1.9 h
Experiment 3 Reporting the Activity Data of This PDC					[5]
Indication	Obesity
Efficacy Data	Vehicle-corrected weight loss rate	23.20%
Administration Time	Once a day for 14 days
Administration Dosage	100 nmol kg-1
Evaluation Method	Over a 14-day treatment period, GLP-1-MK-801 synergistically lowered body weight compared with the dose-matched monotherapies and produced a vehicle-corrected weight loss of 23.2%.
MOA of PDC	Treatment with GLP-1-inactive MK-801 produced no additional weight loss efficacy relative to GLP-1 monotherapy (Fig. 2g-i), indicating that the pronounced weight loss induced by GLP-1-MK-801 is driven by concerted and site-directed pharmacological GLP-1 receptor agonism and NMDA receptor antagonism.
In Vivo Model	DIO mice
Half life period	1.9 h
Experiment 4 Reporting the Activity Data of This PDC					[5]
Indication	Obesity
Efficacy Data	Lean mass rate	8%
Administration Time	Once a day for 14 days
Administration Dosage	100 nmol kg-1
Evaluation Method	GLP-1-MK-801 produced a vehicle-corrected reduction in body fat mass of 45%, accompanied by an 8% loss in lean mass.
MOA of PDC	Treatment with GLP-1-inactive MK-801 produced no additional weight loss efficacy relative to GLP-1 monotherapy (Fig. 2g-i), indicating that the pronounced weight loss induced by GLP-1-MK-801 is driven by concerted and site-directed pharmacological GLP-1 receptor agonism and NMDA receptor antagonism.
In Vivo Model	DIO mice
Half life period	1.9 h
Experiment 5 Reporting the Activity Data of This PDC					[5]
Indication	Obesity
Efficacy Data	Fat mass reduction rate	45%
Administration Time	Once a day for 14 days
Administration Dosage	100 nmol kg-1
Evaluation Method	GLP-1-MK-801 produced a vehicle-corrected reduction in body fat mass of 45%, accompanied by an 8% loss in lean mass.
MOA of PDC	Treatment with GLP-1-inactive MK-801 produced no additional weight loss efficacy relative to GLP-1 monotherapy (Fig. 2g-i), indicating that the pronounced weight loss induced by GLP-1-MK-801 is driven by concerted and site-directed pharmacological GLP-1 receptor agonism and NMDA receptor antagonism.
In Vivo Model	DIO mice
Half life period	1.9 h

Experiment 1 Reporting the Activity Data of This PDC					[6]
Indication	Breast cancer
Efficacy Data	Tumor Growth Inhibition value (TGI)	60% (Day 14)
Administration Time	Injected via tail vein every t hree days
Administration Dosage	8 µmol/kg
MOA of PDC	In the previous study, we focused on the discovery of novel cell targeting peptides, leading to the identification of the preferred HER2-targeting peptide P1 (NPNWGRSWYNQRFK) derived from pertuzumab, with the Kd of 5.70 10-6 M. Here, we designed and synthesized a series of PDCs choosing Camptothecin (CPT) as the payload and P1 as the peptide carrier. We aimed to discover novel PDC molecules with potent inhibitory activity and selectivity targeting HER2 for effective cancer treatment. Conjugate Z8, which balanced the activity and the biosafety, showed good antiproliferative activity (IC50 = 1.91 ± 0.71 M), comparable to that of CPT (IC50 = 2.10 ± 1.34 M), against NCI-N87 cells. Moreover, subsequent verification tests indicated that Z8 operated through a mechanism similar to CPT, but the results from in vivo anti-tumor activity assay suggested that Z8 exhibited greater safety compared to CPT to some extent. These findings highlight Z8 as a promising candidate for further investigation in the treatment of HER2-positive tumors.    Click to Show/Hide
Description	To study the in vivo anti-tumor activity of Z8, SK-BR-3 tumor bearing nude mice were administered once every three days. As shown in Fig. 11A-11D, CPT and Z8 demonstrated significant inhibitory effects on SK-BR-3 tumor growth, with the Z8 group exhibiting more pronounced inhibition compared to CPT. Notably, the average tumor volume and weight in the Z8 group were the smallest among all groups, including the control and CPT groups. Moreover, to further study the toxicity of CPT and Z8, serums of mice were collected for blood biochemical analysis. As shown in Fig. 11E-H, treatment with both CPT and Z8 resulted in increased levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), but the effect of Z8 group was relatively low, indicating that Z8 was safer than CPT in vivo. Furthermore, in CPT group, urea nitrogen levels increased significantly, whereas no significant change was observed in the Z8 group, indicating potential renal function impairment following CPT treatment, whereas Z8 showed better safety in this regard. Additionally, the levels of alkaline phosphatase (ALP), total protein (TP), albumin (ALB), globulin (GLOB) and creatinine (CREA) did not significantly differ between the control group and the treatment groups. Finally, compared to the control group, histological examination using H&E staining (Hematoxylin and Eosin staining) of the major organs in the treatment group did not reveal obvious cell necrosis and inflammatory cell infiltration, indicating that Z8 would not bring additional toxicity.    Click to Show/Hide
In Vivo Model	SK-BR-3 tumor-bearing nude mice xenograft model.
In Vitro Model	Breast adenocarcinoma	SK-BR-3 cell		CVCL_0033

Experiment 1 Reporting the Activity Data of This PDC					[6]
Indication	Breast cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	1.04 ± 0.24 µM
Administration Time	48 h
Evaluation Method	CCK-8 assay
MOA of PDC	In the previous study, we focused on the discovery of novel cell targeting peptides, leading to the identification of the preferred HER2-targeting peptide P1 (NPNWGRSWYNQRFK) derived from pertuzumab, with the Kd of 5.70 10-6 M. Here, we designed and synthesized a series of PDCs choosing Camptothecin (CPT) as the payload and P1 as the peptide carrier. We aimed to discover novel PDC molecules with potent inhibitory activity and selectivity targeting HER2 for effective cancer treatment. Conjugate Z8, which balanced the activity and the biosafety, showed good antiproliferative activity (IC50 = 1.91 ± 0.71 M), comparable to that of CPT (IC50 = 2.10 ± 1.34 M), against NCI-N87 cells. Moreover, subsequent verification tests indicated that Z8 operated through a mechanism similar to CPT, but the results from in vivo anti-tumor activity assay suggested that Z8 exhibited greater safety compared to CPT to some extent. These findings highlight Z8 as a promising candidate for further investigation in the treatment of HER2-positive tumors.    Click to Show/Hide
Description	To evaluated the in vitro anti-tumor activity of the conjugates, HER2-positive SK-BR-3, NCI-N87, HER2-negative MDA-MB-231 and normal MCF-10A cells were selected. As shown in Table 2, nearly all conjugates exhibited obvious anti-proliferation activity against SK-BR-3 cells, while most of the conjugates, except for Z6 and Z9, demonstrated different degrees of inhibitory activity on NCI-N87 cells. Taken together, Z8 and Z11 exhibited superior anti-tumor activity. The antiproliferative activity of Z8 against NCI-N87 cells (IC50 = 1.91 ± 0.71 uM) was comparable to that of CPT (IC50 = 2.10 ± 1.34 uM), while the antiproliferative activity of Z11 against SK-BR-3 cells (IC50 = 0.41 ± 0.13 uM) was comparable to that of CPT (IC50 = 0.26 ± 0.06 uM). Additionally, the IC50 values of these two conjugates against MDA-MB-231 and MCF-10A cells were significantly increased, showing their ability to selectively inhibit tumor cells. Based on the above results, Z8 and Z11, which exhibited better activity and apparent selectivity, were selected for subsequent studies.    Click to Show/Hide
In Vitro Model	Breast adenocarcinoma	SK-BR-3 cell		CVCL_0033
Experiment 2 Reporting the Activity Data of This PDC					[6]
Indication	Gastric cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	1.91 ± 0.71 µM
Administration Time	48 h
Evaluation Method	CCK-8 assay
MOA of PDC	In the previous study, we focused on the discovery of novel cell targeting peptides, leading to the identification of the preferred HER2-targeting peptide P1 (NPNWGRSWYNQRFK) derived from pertuzumab, with the Kd of 5.70 10-6 M. Here, we designed and synthesized a series of PDCs choosing Camptothecin (CPT) as the payload and P1 as the peptide carrier. We aimed to discover novel PDC molecules with potent inhibitory activity and selectivity targeting HER2 for effective cancer treatment. Conjugate Z8, which balanced the activity and the biosafety, showed good antiproliferative activity (IC50 = 1.91 ± 0.71 M), comparable to that of CPT (IC50 = 2.10 ± 1.34 M), against NCI-N87 cells. Moreover, subsequent verification tests indicated that Z8 operated through a mechanism similar to CPT, but the results from in vivo anti-tumor activity assay suggested that Z8 exhibited greater safety compared to CPT to some extent. These findings highlight Z8 as a promising candidate for further investigation in the treatment of HER2-positive tumors.    Click to Show/Hide
Description	To evaluated the in vitro anti-tumor activity of the conjugates, HER2-positive SK-BR-3, NCI-N87, HER2-negative MDA-MB-231 and normal MCF-10A cells were selected. As shown in Table 2, nearly all conjugates exhibited obvious anti-proliferation activity against SK-BR-3 cells, while most of the conjugates, except for Z6 and Z9, demonstrated different degrees of inhibitory activity on NCI-N87 cells. Taken together, Z8 and Z11 exhibited superior anti-tumor activity. The antiproliferative activity of Z8 against NCI-N87 cells (IC50 = 1.91 ± 0.71 uM) was comparable to that of CPT (IC50 = 2.10 ± 1.34 uM), while the antiproliferative activity of Z11 against SK-BR-3 cells (IC50 = 0.41 ± 0.13 uM) was comparable to that of CPT (IC50 = 0.26 ± 0.06 uM). Additionally, the IC50 values of these two conjugates against MDA-MB-231 and MCF-10A cells were significantly increased, showing their ability to selectively inhibit tumor cells. Based on the above results, Z8 and Z11, which exhibited better activity and apparent selectivity, were selected for subsequent studies.    Click to Show/Hide
In Vitro Model	Gastric tubular adenocarcinoma	NCI-N87 cell		CVCL_1603
Experiment 3 Reporting the Activity Data of This PDC					[6]
Indication	Breast cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	13.49 ± 3.59 µM
Administration Time	48 h
Evaluation Method	CCK-8 assay
MOA of PDC	In the previous study, we focused on the discovery of novel cell targeting peptides, leading to the identification of the preferred HER2-targeting peptide P1 (NPNWGRSWYNQRFK) derived from pertuzumab, with the Kd of 5.70 10-6 M. Here, we designed and synthesized a series of PDCs choosing Camptothecin (CPT) as the payload and P1 as the peptide carrier. We aimed to discover novel PDC molecules with potent inhibitory activity and selectivity targeting HER2 for effective cancer treatment. Conjugate Z8, which balanced the activity and the biosafety, showed good antiproliferative activity (IC50 = 1.91 ± 0.71 M), comparable to that of CPT (IC50 = 2.10 ± 1.34 M), against NCI-N87 cells. Moreover, subsequent verification tests indicated that Z8 operated through a mechanism similar to CPT, but the results from in vivo anti-tumor activity assay suggested that Z8 exhibited greater safety compared to CPT to some extent. These findings highlight Z8 as a promising candidate for further investigation in the treatment of HER2-positive tumors.    Click to Show/Hide
Description	To evaluated the in vitro anti-tumor activity of the conjugates, HER2-positive SK-BR-3, NCI-N87, HER2-negative MDA-MB-231 and normal MCF-10A cells were selected. As shown in Table 2, nearly all conjugates exhibited obvious anti-proliferation activity against SK-BR-3 cells, while most of the conjugates, except for Z6 and Z9, demonstrated different degrees of inhibitory activity on NCI-N87 cells. Taken together, Z8 and Z11 exhibited superior anti-tumor activity. The antiproliferative activity of Z8 against NCI-N87 cells (IC50 = 1.91 ± 0.71 uM) was comparable to that of CPT (IC50 = 2.10 ± 1.34 uM), while the antiproliferative activity of Z11 against SK-BR-3 cells (IC50 = 0.41 ± 0.13 uM) was comparable to that of CPT (IC50 = 0.26 ± 0.06 uM). Additionally, the IC50 values of these two conjugates against MDA-MB-231 and MCF-10A cells were significantly increased, showing their ability to selectively inhibit tumor cells. Based on the above results, Z8 and Z11, which exhibited better activity and apparent selectivity, were selected for subsequent studies.    Click to Show/Hide
In Vitro Model	Breast adenocarcinoma	MDA-MB-231 cell		CVCL_0062
Experiment 4 Reporting the Activity Data of This PDC					[6]
Indication	Breast cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	26.34 ± 1.08 µM
Administration Time	48 h
Evaluation Method	CCK-8 assay
MOA of PDC	In the previous study, we focused on the discovery of novel cell targeting peptides, leading to the identification of the preferred HER2-targeting peptide P1 (NPNWGRSWYNQRFK) derived from pertuzumab, with the Kd of 5.70 10-6 M. Here, we designed and synthesized a series of PDCs choosing Camptothecin (CPT) as the payload and P1 as the peptide carrier. We aimed to discover novel PDC molecules with potent inhibitory activity and selectivity targeting HER2 for effective cancer treatment. Conjugate Z8, which balanced the activity and the biosafety, showed good antiproliferative activity (IC50 = 1.91 ± 0.71 M), comparable to that of CPT (IC50 = 2.10 ± 1.34 M), against NCI-N87 cells. Moreover, subsequent verification tests indicated that Z8 operated through a mechanism similar to CPT, but the results from in vivo anti-tumor activity assay suggested that Z8 exhibited greater safety compared to CPT to some extent. These findings highlight Z8 as a promising candidate for further investigation in the treatment of HER2-positive tumors.    Click to Show/Hide
Description	To evaluated the in vitro anti-tumor activity of the conjugates, HER2-positive SK-BR-3, NCI-N87, HER2-negative MDA-MB-231 and normal MCF-10A cells were selected. As shown in Table 2, nearly all conjugates exhibited obvious anti-proliferation activity against SK-BR-3 cells, while most of the conjugates, except for Z6 and Z9, demonstrated different degrees of inhibitory activity on NCI-N87 cells. Taken together, Z8 and Z11 exhibited superior anti-tumor activity. The antiproliferative activity of Z8 against NCI-N87 cells (IC50 = 1.91 ± 0.71 uM) was comparable to that of CPT (IC50 = 2.10 ± 1.34 uM), while the antiproliferative activity of Z11 against SK-BR-3 cells (IC50 = 0.41 ± 0.13 uM) was comparable to that of CPT (IC50 = 0.26 ± 0.06 uM). Additionally, the IC50 values of these two conjugates against MDA-MB-231 and MCF-10A cells were significantly increased, showing their ability to selectively inhibit tumor cells. Based on the above results, Z8 and Z11, which exhibited better activity and apparent selectivity, were selected for subsequent studies.    Click to Show/Hide
In Vitro Model	Normal	MCF-10A cell		CVCL_0598
Experiment 5 Reporting the Activity Data of This PDC					[6]
Indication	Breast cancer
Efficacy Data	Apoptosis rate	27.10%
Administration Time	24 h
Administration Dosage	1 µM
Evaluation Method	Flow cytometry assay
MOA of PDC	In the previous study, we focused on the discovery of novel cell targeting peptides, leading to the identification of the preferred HER2-targeting peptide P1 (NPNWGRSWYNQRFK) derived from pertuzumab, with the Kd of 5.70 10-6 M. Here, we designed and synthesized a series of PDCs choosing Camptothecin (CPT) as the payload and P1 as the peptide carrier. We aimed to discover novel PDC molecules with potent inhibitory activity and selectivity targeting HER2 for effective cancer treatment. Conjugate Z8, which balanced the activity and the biosafety, showed good antiproliferative activity (IC50 = 1.91 ± 0.71 M), comparable to that of CPT (IC50 = 2.10 ± 1.34 M), against NCI-N87 cells. Moreover, subsequent verification tests indicated that Z8 operated through a mechanism similar to CPT, but the results from in vivo anti-tumor activity assay suggested that Z8 exhibited greater safety compared to CPT to some extent. These findings highlight Z8 as a promising candidate for further investigation in the treatment of HER2-positive tumors.    Click to Show/Hide
Description	The proapoptotic activity of Z8 was studied by flow cytometry combined with Annexin V-FITC/PI double staining. As shown in Fig. 4, after treatment with 1 uM Z8, the apoptosis rate of SK-BR-3 cells was approximately 27.1 % which increased to 41.1 % following treatment with 2 uM Z8 (4.4 % in the blank control group and 55.8 % in CPT group. These findings suggest that Z8 can significantly induce apoptosis in a dose-dependent manner.    Click to Show/Hide
In Vitro Model	Breast adenocarcinoma	SK-BR-3 cell		CVCL_0033
Experiment 6 Reporting the Activity Data of This PDC					[6]
Indication	Breast cancer
Efficacy Data	Apoptosis rate	41.10%
Administration Time	24 h
Administration Dosage	2 µM
Evaluation Method	Flow cytometry assay
MOA of PDC	In the previous study, we focused on the discovery of novel cell targeting peptides, leading to the identification of the preferred HER2-targeting peptide P1 (NPNWGRSWYNQRFK) derived from pertuzumab, with the Kd of 5.70 10-6 M. Here, we designed and synthesized a series of PDCs choosing Camptothecin (CPT) as the payload and P1 as the peptide carrier. We aimed to discover novel PDC molecules with potent inhibitory activity and selectivity targeting HER2 for effective cancer treatment. Conjugate Z8, which balanced the activity and the biosafety, showed good antiproliferative activity (IC50 = 1.91 ± 0.71 M), comparable to that of CPT (IC50 = 2.10 ± 1.34 M), against NCI-N87 cells. Moreover, subsequent verification tests indicated that Z8 operated through a mechanism similar to CPT, but the results from in vivo anti-tumor activity assay suggested that Z8 exhibited greater safety compared to CPT to some extent. These findings highlight Z8 as a promising candidate for further investigation in the treatment of HER2-positive tumors.    Click to Show/Hide
Description	The proapoptotic activity of Z8 was studied by flow cytometry combined with Annexin V-FITC/PI double staining. As shown in Fig. 4, after treatment with 1 uM Z8, the apoptosis rate of SK-BR-3 cells was approximately 27.1 % which increased to 41.1 % following treatment with 2 uM Z8 (4.4 % in the blank control group and 55.8 % in CPT group. These findings suggest that Z8 can significantly induce apoptosis in a dose-dependent manner.    Click to Show/Hide
In Vitro Model	Breast adenocarcinoma	SK-BR-3 cell		CVCL_0033

Experiment 1 Reporting the Activity Data of This PDC					[7]
Indication	Solid tumor
Efficacy Data	Tumor growth inhibition value (TGI)	67.10%
Administration Time	Every two days for 2 weeks
Administration Dosage	8 μmol/kg
MOA of PDC	LHRH, also named gonadotropin-releasing hormone (GnRH), is an endogenous peptide agonist (primary sequence: pGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH2) released from hypothalamus. LHRH-R (LHRH receptor), a member of the G protein-coupled receptor family, is overexpressed in various tumor types, while their expression in the corresponding normal tissues, apart from pituitary cells, is comparatively low. Given this, we chose LHRH as the TTP component of MSCPTP (TTP-CPP peptide). In this investigation, we combined LHRH (as the TTP part), peptide PLGLAG, T2 (as the CPP part) and cysteine (as linker binding site) into an MSCPTP named LT-1. Then PTX was conjugated with LT-1 via a GSH-cleavable module to produce the smart PDC, namely LTP-1 (TTP-CPP-PTX conjugate). In vitro, LTP-1 exhibited selective and stronger cytotoxicity than PTX against LHRH-R-positive tumor cells with little effect on normal cells. In vivo, LTP-1 was highly effective in suppressing tumor growth in an MCF-7 xenograft mouse model. Additional experiments on both cellular and molecular levels were carried out to unravel the possible antitumor mechanism of action of LTP-1.    Click to Show/Hide
Description	At the dose of 8 μmol/kg, LTP-1 decreased the tumor volume and tumor weight by 77.2% and 67.1%.
In Vivo Model	MCF-7 xenograft mice.
Experiment 2 Reporting the Activity Data of This PDC					[7]
Indication	Solid tumor
Efficacy Data	Tumor growth inhibition value (TGI)	77.20%
Administration Time	Every two days for 2 weeks
Administration Dosage	8 μmol/kg
Evaluation Method	Tumor volume detection
MOA of PDC	LHRH, also named gonadotropin-releasing hormone (GnRH), is an endogenous peptide agonist (primary sequence: pGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH2) released from hypothalamus. LHRH-R (LHRH receptor), a member of the G protein-coupled receptor family, is overexpressed in various tumor types, while their expression in the corresponding normal tissues, apart from pituitary cells, is comparatively low. Given this, we chose LHRH as the TTP component of MSCPTP (TTP-CPP peptide). In this investigation, we combined LHRH (as the TTP part), peptide PLGLAG, T2 (as the CPP part) and cysteine (as linker binding site) into an MSCPTP named LT-1. Then PTX was conjugated with LT-1 via a GSH-cleavable module to produce the smart PDC, namely LTP-1 (TTP-CPP-PTX conjugate). In vitro, LTP-1 exhibited selective and stronger cytotoxicity than PTX against LHRH-R-positive tumor cells with little effect on normal cells. In vivo, LTP-1 was highly effective in suppressing tumor growth in an MCF-7 xenograft mouse model. Additional experiments on both cellular and molecular levels were carried out to unravel the possible antitumor mechanism of action of LTP-1.    Click to Show/Hide
Description	At the dose of 8 μmol/kg, LTP-1 decreased the tumor volume and tumor weight by 77.2% and 67.1%.
In Vivo Model	MCF-7 xenograft mice.
Experiment 3 Reporting the Activity Data of This PDC					[7]
Indication	Solid tumor
Efficacy Data	Tumor growth inhibition value (TGI)	83.40%
Administration Time	Every two days for 2 weeks
Administration Dosage	12 μmol/kg
MOA of PDC	LHRH, also named gonadotropin-releasing hormone (GnRH), is an endogenous peptide agonist (primary sequence: pGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH2) released from hypothalamus. LHRH-R (LHRH receptor), a member of the G protein-coupled receptor family, is overexpressed in various tumor types, while their expression in the corresponding normal tissues, apart from pituitary cells, is comparatively low. Given this, we chose LHRH as the TTP component of MSCPTP (TTP-CPP peptide). In this investigation, we combined LHRH (as the TTP part), peptide PLGLAG, T2 (as the CPP part) and cysteine (as linker binding site) into an MSCPTP named LT-1. Then PTX was conjugated with LT-1 via a GSH-cleavable module to produce the smart PDC, namely LTP-1 (TTP-CPP-PTX conjugate). In vitro, LTP-1 exhibited selective and stronger cytotoxicity than PTX against LHRH-R-positive tumor cells with little effect on normal cells. In vivo, LTP-1 was highly effective in suppressing tumor growth in an MCF-7 xenograft mouse model. Additional experiments on both cellular and molecular levels were carried out to unravel the possible antitumor mechanism of action of LTP-1.    Click to Show/Hide
Description	At the dose of 12 μmol/kg, LTP-1 decreased the tumor volume and tumor weight by 90.1% and 83.4%, respectively.
In Vivo Model	MCF-7 xenograft mice.
Experiment 4 Reporting the Activity Data of This PDC					[7]
Indication	Solid tumor
Efficacy Data	Tumor growth inhibition value (TGI)	90.10%
Administration Time	Every two days for 2 weeks
Administration Dosage	12 μmol/kg
Evaluation Method	Tumor volume detection
MOA of PDC	LHRH, also named gonadotropin-releasing hormone (GnRH), is an endogenous peptide agonist (primary sequence: pGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH2) released from hypothalamus. LHRH-R (LHRH receptor), a member of the G protein-coupled receptor family, is overexpressed in various tumor types, while their expression in the corresponding normal tissues, apart from pituitary cells, is comparatively low. Given this, we chose LHRH as the TTP component of MSCPTP (TTP-CPP peptide). In this investigation, we combined LHRH (as the TTP part), peptide PLGLAG, T2 (as the CPP part) and cysteine (as linker binding site) into an MSCPTP named LT-1. Then PTX was conjugated with LT-1 via a GSH-cleavable module to produce the smart PDC, namely LTP-1 (TTP-CPP-PTX conjugate). In vitro, LTP-1 exhibited selective and stronger cytotoxicity than PTX against LHRH-R-positive tumor cells with little effect on normal cells. In vivo, LTP-1 was highly effective in suppressing tumor growth in an MCF-7 xenograft mouse model. Additional experiments on both cellular and molecular levels were carried out to unravel the possible antitumor mechanism of action of LTP-1.    Click to Show/Hide
Description	At the dose of 12 μmol/kg, LTP-1 decreased the tumor volume and tumor weight by 90.1% and 83.4%, respectively.
In Vivo Model	MCF-7 xenograft mice.

Experiment 1 Reporting the Activity Data of This PDC					[7]
Indication	Solid tumor
Efficacy Data	Half maximal inhibitory concentration (IC50)	3.8 ± 0.3 nM
Evaluation Method	MTT assay
MOA of PDC	LHRH, also named gonadotropin-releasing hormone (GnRH), is an endogenous peptide agonist (primary sequence: pGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH2) released from hypothalamus. LHRH-R (LHRH receptor), a member of the G protein-coupled receptor family, is overexpressed in various tumor types, while their expression in the corresponding normal tissues, apart from pituitary cells, is comparatively low. Given this, we chose LHRH as the TTP component of MSCPTP (TTP-CPP peptide). In this investigation, we combined LHRH (as the TTP part), peptide PLGLAG, T2 (as the CPP part) and cysteine (as linker binding site) into an MSCPTP named LT-1. Then PTX was conjugated with LT-1 via a GSH-cleavable module to produce the smart PDC, namely LTP-1 (TTP-CPP-PTX conjugate). In vitro, LTP-1 exhibited selective and stronger cytotoxicity than PTX against LHRH-R-positive tumor cells with little effect on normal cells. In vivo, LTP-1 was highly effective in suppressing tumor growth in an MCF-7 xenograft mouse model. Additional experiments on both cellular and molecular levels were carried out to unravel the possible antitumor mechanism of action of LTP-1.    Click to Show/Hide
Description	LTP-1 exhibited greater anti-proliferative effects (IC50s of 3.8-20.3 nM) than PTX (IC50s of 6.6-28.6 nM) against most cancer cells except Hela, and less cytotoxicity to normal cells (IC50s of >80 nM and 66.0 nM for NCM460 and HEK-293, respectively). Thus, LTP-1 displayed not only enhanced anti-proliferative activity, but also higher selectivity for cancer cells over normal cells. It is also worthy of note that LTP-1 showed much higher activity against the paclitaxel-resistant A2780/PTX cells with an IC50 of 0.8 μM, as compared to PTX which is essentially inactive (IC50 = 23.9 μM). Hemolysis assay further testified that LTP-1 presented weak hemolytic activity even at a concentration up to 80 μM (as illustrated in Fig. 4a).    Click to Show/Hide
In Vitro Model	Invasive breast carcinoma	MCF-7 cell		CVCL_0031
Experiment 2 Reporting the Activity Data of This PDC					[7]
Indication	Solid tumor
Efficacy Data	Half maximal inhibitory concentration (IC50)	5.6 ± 0.2 nM
Evaluation Method	MTT assay
MOA of PDC	LHRH, also named gonadotropin-releasing hormone (GnRH), is an endogenous peptide agonist (primary sequence: pGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH2) released from hypothalamus. LHRH-R (LHRH receptor), a member of the G protein-coupled receptor family, is overexpressed in various tumor types, while their expression in the corresponding normal tissues, apart from pituitary cells, is comparatively low. Given this, we chose LHRH as the TTP component of MSCPTP (TTP-CPP peptide). In this investigation, we combined LHRH (as the TTP part), peptide PLGLAG, T2 (as the CPP part) and cysteine (as linker binding site) into an MSCPTP named LT-1. Then PTX was conjugated with LT-1 via a GSH-cleavable module to produce the smart PDC, namely LTP-1 (TTP-CPP-PTX conjugate). In vitro, LTP-1 exhibited selective and stronger cytotoxicity than PTX against LHRH-R-positive tumor cells with little effect on normal cells. In vivo, LTP-1 was highly effective in suppressing tumor growth in an MCF-7 xenograft mouse model. Additional experiments on both cellular and molecular levels were carried out to unravel the possible antitumor mechanism of action of LTP-1.    Click to Show/Hide
Description	LTP-1 exhibited greater anti-proliferative effects (IC50s of 3.8-20.3 nM) than PTX (IC50s of 6.6-28.6 nM) against most cancer cells except Hela, and less cytotoxicity to normal cells (IC50s of >80 nM and 66.0 nM for NCM460 and HEK-293, respectively). Thus, LTP-1 displayed not only enhanced anti-proliferative activity, but also higher selectivity for cancer cells over normal cells. It is also worthy of note that LTP-1 showed much higher activity against the paclitaxel-resistant A2780/PTX cells with an IC50 of 0.8 μM, as compared to PTX which is essentially inactive (IC50 = 23.9 μM). Hemolysis assay further testified that LTP-1 presented weak hemolytic activity even at a concentration up to 80 μM (as illustrated in Fig. 4a).    Click to Show/Hide
In Vitro Model	Colon adenocarcinoma	HT-29 cell		CVCL_0320
Experiment 3 Reporting the Activity Data of This PDC					[7]
Indication	Solid tumor
Efficacy Data	Half maximal inhibitory concentration (IC50)	8.3 ± 0.5 nM
Evaluation Method	MTT assay
MOA of PDC	LHRH, also named gonadotropin-releasing hormone (GnRH), is an endogenous peptide agonist (primary sequence: pGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH2) released from hypothalamus. LHRH-R (LHRH receptor), a member of the G protein-coupled receptor family, is overexpressed in various tumor types, while their expression in the corresponding normal tissues, apart from pituitary cells, is comparatively low. Given this, we chose LHRH as the TTP component of MSCPTP (TTP-CPP peptide). In this investigation, we combined LHRH (as the TTP part), peptide PLGLAG, T2 (as the CPP part) and cysteine (as linker binding site) into an MSCPTP named LT-1. Then PTX was conjugated with LT-1 via a GSH-cleavable module to produce the smart PDC, namely LTP-1 (TTP-CPP-PTX conjugate). In vitro, LTP-1 exhibited selective and stronger cytotoxicity than PTX against LHRH-R-positive tumor cells with little effect on normal cells. In vivo, LTP-1 was highly effective in suppressing tumor growth in an MCF-7 xenograft mouse model. Additional experiments on both cellular and molecular levels were carried out to unravel the possible antitumor mechanism of action of LTP-1.    Click to Show/Hide
Description	LTP-1 exhibited greater anti-proliferative effects (IC50s of 3.8-20.3 nM) than PTX (IC50s of 6.6-28.6 nM) against most cancer cells except Hela, and less cytotoxicity to normal cells (IC50s of >80 nM and 66.0 nM for NCM460 and HEK-293, respectively). Thus, LTP-1 displayed not only enhanced anti-proliferative activity, but also higher selectivity for cancer cells over normal cells. It is also worthy of note that LTP-1 showed much higher activity against the paclitaxel-resistant A2780/PTX cells with an IC50 of 0.8 μM, as compared to PTX which is essentially inactive (IC50 = 23.9 μM). Hemolysis assay further testified that LTP-1 presented weak hemolytic activity even at a concentration up to 80 μM (as illustrated in Fig. 4a).    Click to Show/Hide
In Vitro Model	Ovarian endometrioid adenocarcinoma	A2780 cell		CVCL_0134
Experiment 4 Reporting the Activity Data of This PDC					[7]
Indication	Solid tumor
Efficacy Data	Half maximal inhibitory concentration (IC50)	20.3 ± 3.3 nM
Evaluation Method	MTT assay
MOA of PDC	LHRH, also named gonadotropin-releasing hormone (GnRH), is an endogenous peptide agonist (primary sequence: pGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH2) released from hypothalamus. LHRH-R (LHRH receptor), a member of the G protein-coupled receptor family, is overexpressed in various tumor types, while their expression in the corresponding normal tissues, apart from pituitary cells, is comparatively low. Given this, we chose LHRH as the TTP component of MSCPTP (TTP-CPP peptide). In this investigation, we combined LHRH (as the TTP part), peptide PLGLAG, T2 (as the CPP part) and cysteine (as linker binding site) into an MSCPTP named LT-1. Then PTX was conjugated with LT-1 via a GSH-cleavable module to produce the smart PDC, namely LTP-1 (TTP-CPP-PTX conjugate). In vitro, LTP-1 exhibited selective and stronger cytotoxicity than PTX against LHRH-R-positive tumor cells with little effect on normal cells. In vivo, LTP-1 was highly effective in suppressing tumor growth in an MCF-7 xenograft mouse model. Additional experiments on both cellular and molecular levels were carried out to unravel the possible antitumor mechanism of action of LTP-1.    Click to Show/Hide
Description	LTP-1 exhibited greater anti-proliferative effects (IC50s of 3.8-20.3 nM) than PTX (IC50s of 6.6-28.6 nM) against most cancer cells except Hela, and less cytotoxicity to normal cells (IC50s of >80 nM and 66.0 nM for NCM460 and HEK-293, respectively). Thus, LTP-1 displayed not only enhanced anti-proliferative activity, but also higher selectivity for cancer cells over normal cells. It is also worthy of note that LTP-1 showed much higher activity against the paclitaxel-resistant A2780/PTX cells with an IC50 of 0.8 μM, as compared to PTX which is essentially inactive (IC50 = 23.9 μM). Hemolysis assay further testified that LTP-1 presented weak hemolytic activity even at a concentration up to 80 μM (as illustrated in Fig. 4a).    Click to Show/Hide
In Vitro Model	Human papillomavirus-related cervical adenocarcinoma	HeLa cell		CVCL_0030
Experiment 5 Reporting the Activity Data of This PDC					[7]
Indication	Solid tumor
Efficacy Data	Half maximal inhibitory concentration (IC50)	66.0 ± 8.3 nM
Evaluation Method	MTT assay
MOA of PDC	LHRH, also named gonadotropin-releasing hormone (GnRH), is an endogenous peptide agonist (primary sequence: pGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH2) released from hypothalamus. LHRH-R (LHRH receptor), a member of the G protein-coupled receptor family, is overexpressed in various tumor types, while their expression in the corresponding normal tissues, apart from pituitary cells, is comparatively low. Given this, we chose LHRH as the TTP component of MSCPTP (TTP-CPP peptide). In this investigation, we combined LHRH (as the TTP part), peptide PLGLAG, T2 (as the CPP part) and cysteine (as linker binding site) into an MSCPTP named LT-1. Then PTX was conjugated with LT-1 via a GSH-cleavable module to produce the smart PDC, namely LTP-1 (TTP-CPP-PTX conjugate). In vitro, LTP-1 exhibited selective and stronger cytotoxicity than PTX against LHRH-R-positive tumor cells with little effect on normal cells. In vivo, LTP-1 was highly effective in suppressing tumor growth in an MCF-7 xenograft mouse model. Additional experiments on both cellular and molecular levels were carried out to unravel the possible antitumor mechanism of action of LTP-1.    Click to Show/Hide
Description	LTP-1 exhibited greater anti-proliferative effects (IC50s of 3.8-20.3 nM) than PTX (IC50s of 6.6-28.6 nM) against most cancer cells except Hela, and less cytotoxicity to normal cells (IC50s of >80 nM and 66.0 nM for NCM460 and HEK-293, respectively). Thus, LTP-1 displayed not only enhanced anti-proliferative activity, but also higher selectivity for cancer cells over normal cells. It is also worthy of note that LTP-1 showed much higher activity against the paclitaxel-resistant A2780/PTX cells with an IC50 of 0.8 μM, as compared to PTX which is essentially inactive (IC50 = 23.9 μM). Hemolysis assay further testified that LTP-1 presented weak hemolytic activity even at a concentration up to 80 μM (as illustrated in Fig. 4a).    Click to Show/Hide
In Vitro Model	Normal	HEK293 cell		CVCL_0045
Experiment 6 Reporting the Activity Data of This PDC					[7]
Indication	Solid tumor
Efficacy Data	Half maximal inhibitory concentration (IC50)	> 80.0 nM
Evaluation Method	MTT assay
MOA of PDC	LHRH, also named gonadotropin-releasing hormone (GnRH), is an endogenous peptide agonist (primary sequence: pGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH2) released from hypothalamus. LHRH-R (LHRH receptor), a member of the G protein-coupled receptor family, is overexpressed in various tumor types, while their expression in the corresponding normal tissues, apart from pituitary cells, is comparatively low. Given this, we chose LHRH as the TTP component of MSCPTP (TTP-CPP peptide). In this investigation, we combined LHRH (as the TTP part), peptide PLGLAG, T2 (as the CPP part) and cysteine (as linker binding site) into an MSCPTP named LT-1. Then PTX was conjugated with LT-1 via a GSH-cleavable module to produce the smart PDC, namely LTP-1 (TTP-CPP-PTX conjugate). In vitro, LTP-1 exhibited selective and stronger cytotoxicity than PTX against LHRH-R-positive tumor cells with little effect on normal cells. In vivo, LTP-1 was highly effective in suppressing tumor growth in an MCF-7 xenograft mouse model. Additional experiments on both cellular and molecular levels were carried out to unravel the possible antitumor mechanism of action of LTP-1.    Click to Show/Hide
Description	LTP-1 exhibited greater anti-proliferative effects (IC50s of 3.8-20.3 nM) than PTX (IC50s of 6.6-28.6 nM) against most cancer cells except Hela, and less cytotoxicity to normal cells (IC50s of >80 nM and 66.0 nM for NCM460 and HEK-293, respectively). Thus, LTP-1 displayed not only enhanced anti-proliferative activity, but also higher selectivity for cancer cells over normal cells. It is also worthy of note that LTP-1 showed much higher activity against the paclitaxel-resistant A2780/PTX cells with an IC50 of 0.8 μM, as compared to PTX which is essentially inactive (IC50 = 23.9 μM). Hemolysis assay further testified that LTP-1 presented weak hemolytic activity even at a concentration up to 80 μM (as illustrated in Fig. 4a).    Click to Show/Hide
In Vitro Model	Normal	NCM460 cell		CVCL_0460
Experiment 7 Reporting the Activity Data of This PDC					[7]
Indication	Solid tumor
Efficacy Data	Half maximal inhibitory concentration (IC50)	800 ± 100 nM
Evaluation Method	MTT assay
MOA of PDC	LHRH, also named gonadotropin-releasing hormone (GnRH), is an endogenous peptide agonist (primary sequence: pGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH2) released from hypothalamus. LHRH-R (LHRH receptor), a member of the G protein-coupled receptor family, is overexpressed in various tumor types, while their expression in the corresponding normal tissues, apart from pituitary cells, is comparatively low. Given this, we chose LHRH as the TTP component of MSCPTP (TTP-CPP peptide). In this investigation, we combined LHRH (as the TTP part), peptide PLGLAG, T2 (as the CPP part) and cysteine (as linker binding site) into an MSCPTP named LT-1. Then PTX was conjugated with LT-1 via a GSH-cleavable module to produce the smart PDC, namely LTP-1 (TTP-CPP-PTX conjugate). In vitro, LTP-1 exhibited selective and stronger cytotoxicity than PTX against LHRH-R-positive tumor cells with little effect on normal cells. In vivo, LTP-1 was highly effective in suppressing tumor growth in an MCF-7 xenograft mouse model. Additional experiments on both cellular and molecular levels were carried out to unravel the possible antitumor mechanism of action of LTP-1.    Click to Show/Hide
Description	LTP-1 exhibited greater anti-proliferative effects (IC50s of 3.8-20.3 nM) than PTX (IC50s of 6.6-28.6 nM) against most cancer cells except Hela, and less cytotoxicity to normal cells (IC50s of >80 nM and 66.0 nM for NCM460 and HEK-293, respectively). Thus, LTP-1 displayed not only enhanced anti-proliferative activity, but also higher selectivity for cancer cells over normal cells. It is also worthy of note that LTP-1 showed much higher activity against the paclitaxel-resistant A2780/PTX cells with an IC50 of 0.8 μM, as compared to PTX which is essentially inactive (IC50 = 23.9 μM). Hemolysis assay further testified that LTP-1 presented weak hemolytic activity even at a concentration up to 80 μM (as illustrated in Fig. 4a).    Click to Show/Hide
In Vitro Model	Ovarian endometrioid adenocarcinoma	A2780/PTX cell		CVCL_C0D6

Experiment 1 Reporting the Activity Data of This PDC					[8]
Indication	Tumor
Efficacy Data	Tumer volume	1800 mm3
Administration Time	Given every other day for a total of five times
Administration Dosage	400 µg/kg (calculated by free DM1)
Description	It was demonstrated here, RCCD@NPs and RSSD@NPs exhibited significantly better tumor-growth inhibition compared with that of the free DM1, QCCD@NPs or QSSD@NPs (Figure (Figure4A).4A). RSSD@NPs showed the most suppression on B16 tumor up to 25 days, and resulted in a tumor volume 4 times smaller than the saline group at the end of the experiment. The final tumor volumes in various nano-DDS groups ranked from the greatest to the least: QCCD@NPs, QSSD@NPs>DM1>RCCD@NPs>RSSD@NPs. Almost the same trends were found in terms of tumor weight and tumor size (Figures (Figures4B,4B, C).    Click to Show/Hide

Experiment 1 Reporting the Activity Data of This PDC					[8]
Indication	Tumor
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	20.42 ± 4.70 nM
Administration Time	48 h
Description	The IC50 of QCCD@NPs and QSSD@NPs (389.05±75.25 nM and 245.47±37.54 nM) on B16 cells seemed much higher than the values of RCCD@NPs and RSSD@NPs (102.33±38.92 nM and 21.38±4.32 nM). The cytotoxicity of APDC@NPs on HUVEC showed a similar pattern.
In Vitro Model	Normal	Human umbilical vein endothelial cell		Homo sapiens
Experiment 2 Reporting the Activity Data of This PDC					[8]
Indication	Tumor
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	21.38 ± 4.32 nM
Administration Time	48 h
Description	The IC50 of QCCD@NPs and QSSD@NPs (389.05±75.25 nM and 245.47±37.54 nM) on B16 cells seemed much higher than the values of RCCD@NPs and RSSD@NPs (102.33±38.92 nM and 21.38±4.32 nM). The cytotoxicity of APDC@NPs on HUVEC showed a similar pattern.
In Vitro Model	Melanoma	B16 cell		CVCL_F936
Experiment 3 Reporting the Activity Data of This PDC					[8]
Indication	Tumor
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	302.00 ± 72.91 nM
Administration Time	48 h
Description	The IC50 of QCCD@NPs and QSSD@NPs (389.05±75.25 nM and 245.47±37.54 nM) on B16 cells seemed much higher than the values of RCCD@NPs and RSSD@NPs (102.33±38.92 nM and 21.38±4.32 nM). The cytotoxicity of APDC@NPs on HUVEC showed a similar pattern.
In Vitro Model	Invasive breast carcinoma	MCF-7 cell		CVCL_0031

Experiment 1 Reporting the Activity Data of This PDC					[8]
Indication	Tumor
Efficacy Data	Tumer volume	5200 mm3
Administration Time	Given every other day for a total of five times
Administration Dosage	400 µg/kg (calculated by free DM1)
Description	It was demonstrated here, RCCD@NPs and RSSD@NPs exhibited significantly better tumor-growth inhibition compared with that of the free DM1, QCCD@NPs or QSSD@NPs (Figure (Figure4A).4A). RSSD@NPs showed the most suppression on B16 tumor up to 25 days, and resulted in a tumor volume 4 times smaller than the saline group at the end of the experiment. The final tumor volumes in various nano-DDS groups ranked from the greatest to the least: QCCD@NPs, QSSD@NPs>DM1>RCCD@NPs>RSSD@NPs. Almost the same trends were found in terms of tumor weight and tumor size (Figures (Figures4B,4B, C).    Click to Show/Hide

Experiment 1 Reporting the Activity Data of This PDC					[8]
Indication	Tumor
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	154.88 ± 31.85 nM
Administration Time	48 h
Description	The IC50 of QCCD@NPs and QSSD@NPs (389.05±75.25 nM and 245.47±37.54 nM) on B16 cells seemed much higher than the values of RCCD@NPs and RSSD@NPs (102.33±38.92 nM and 21.38±4.32 nM). The cytotoxicity of APDC@NPs on HUVEC showed a similar pattern.
In Vitro Model	Normal	Human umbilical vein endothelial cell		Homo sapiens
Experiment 2 Reporting the Activity Data of This PDC					[8]
Indication	Tumor
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	245.47 ± 37.54 nM
Administration Time	48 h
Description	The IC50 of QCCD@NPs and QSSD@NPs (389.05±75.25 nM and 245.47±37.54 nM) on B16 cells seemed much higher than the values of RCCD@NPs and RSSD@NPs (102.33±38.92 nM and 21.38±4.32 nM). The cytotoxicity of APDC@NPs on HUVEC showed a similar pattern.
In Vitro Model	Melanoma	B16 cell		CVCL_F936
Experiment 3 Reporting the Activity Data of This PDC					[8]
Indication	Tumor
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	371.53 ± 94.49 nM
Administration Time	48 h
Description	The IC50 of QCCD@NPs and QSSD@NPs (389.05±75.25 nM and 245.47±37.54 nM) on B16 cells seemed much higher than the values of RCCD@NPs and RSSD@NPs (102.33±38.92 nM and 21.38±4.32 nM). The cytotoxicity of APDC@NPs on HUVEC showed a similar pattern.
In Vitro Model	Invasive breast carcinoma	MCF-7 cell		CVCL_0031

Experiment 1 Reporting the Activity Data of This PDC					[9]
Indication	Tumor
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	205 ± 49 nM
Description	As shown in Table 1, the cytotoxic potency of the tubugi-1-SH was - in case of HT-29 and PC-3 - by factors 5 to 8 higher compared to the entire peptide-toxin conjugate 8. The only slight increase of cytotoxic activity of compound 9 compared to the complete conjugate 8 in Colo320 cells is most likely caused by a generally weak responsiveness of Colo320 cells towards tubugi-1-SH and the entire conjugate tubugi-1-SS-NPY. When compared with HT-29 and PC-3 cells, the IC50 value of tubugi-1-SH is by factor 10 higher in Colo320. Since the membrane passage of tubugi-1-SH is not depending on a NPY receptor, there have to be other explanations for the reduced cytotoxic impact of tubugi-1 and corresponding derivatives in Colo320, rather than the NPY Y1 receptor expression level.    Click to Show/Hide
In Vitro Model	Prostate carcinoma	PC-3 cell		CVCL_0035
Experiment 2 Reporting the Activity Data of This PDC					[9]
Indication	Tumor
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	452 ± 60 nM
Description	As shown in Table 1, the cytotoxic potency of the tubugi-1-SH was - in case of HT-29 and PC-3 - by factors 5 to 8 higher compared to the entire peptide-toxin conjugate 8. The only slight increase of cytotoxic activity of compound 9 compared to the complete conjugate 8 in Colo320 cells is most likely caused by a generally weak responsiveness of Colo320 cells towards tubugi-1-SH and the entire conjugate tubugi-1-SS-NPY. When compared with HT-29 and PC-3 cells, the IC50 value of tubugi-1-SH is by factor 10 higher in Colo320. Since the membrane passage of tubugi-1-SH is not depending on a NPY receptor, there have to be other explanations for the reduced cytotoxic impact of tubugi-1 and corresponding derivatives in Colo320, rather than the NPY Y1 receptor expression level.    Click to Show/Hide
In Vitro Model	Colon cancer	HT29 cell		CVCL_A8EZ
Experiment 3 Reporting the Activity Data of This PDC					[9]
Indication	Tumor
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	706 ± 185 nM
Description	As shown in Table 1, the cytotoxic potency of the tubugi-1-SH was - in case of HT-29 and PC-3 - by factors 5 to 8 higher compared to the entire peptide-toxin conjugate 8. The only slight increase of cytotoxic activity of compound 9 compared to the complete conjugate 8 in Colo320 cells is most likely caused by a generally weak responsiveness of Colo320 cells towards tubugi-1-SH and the entire conjugate tubugi-1-SS-NPY. When compared with HT-29 and PC-3 cells, the IC50 value of tubugi-1-SH is by factor 10 higher in Colo320. Since the membrane passage of tubugi-1-SH is not depending on a NPY receptor, there have to be other explanations for the reduced cytotoxic impact of tubugi-1 and corresponding derivatives in Colo320, rather than the NPY Y1 receptor expression level.    Click to Show/Hide
In Vitro Model	Colon adenocarcinoma	COLO 320 cell		CVCL_1989
Experiment 4 Reporting the Activity Data of This PDC					[9]
Indication	Tumor
Efficacy Data	Cell viability	0%
Administration Time	72 h
Administration Dosage	10 µM
Description	The 72 h treatment is more effective than the 6 h pulse treatment. Notably, in vitro antitumor activities of 8 were found to correlate very good with the hY1R expression levels, as detected by gene expression analyses using RT-qPCR. Both the cytotoxic activity and the hY1R expression level rank in the order SK-N-MC > MDA-MB-468 > MDA-MB-231 > 184B5, what proofs the hY1R-specific and -selective nature of the mode of antitumor action of the designed PDC 8. Importantly, the activity of 8 against the selected normal breast cell line 184B5 is in the same order of magnitude as for the hY1R-deficient tumor cell line (MDA-MB-231), both tested at even higher concentration of the PDC than for the Y1 cell lines.    Click to Show/Hide
In Vitro Model	Askin tumor	SK-N-MC cell		CVCL_0530
Experiment 5 Reporting the Activity Data of This PDC					[9]
Indication	Tumor
Efficacy Data	Cell viability	2%
Administration Time	6 h
Administration Dosage	10 µM
Description	The 72 h treatment is more effective than the 6 h pulse treatment. Notably, in vitro antitumor activities of 8 were found to correlate very good with the hY1R expression levels, as detected by gene expression analyses using RT-qPCR. Both the cytotoxic activity and the hY1R expression level rank in the order SK-N-MC > MDA-MB-468 > MDA-MB-231 > 184B5, what proofs the hY1R-specific and -selective nature of the mode of antitumor action of the designed PDC 8. Importantly, the activity of 8 against the selected normal breast cell line 184B5 is in the same order of magnitude as for the hY1R-deficient tumor cell line (MDA-MB-231), both tested at even higher concentration of the PDC than for the Y1 cell lines.    Click to Show/Hide
In Vitro Model	Askin tumor	SK-N-MC cell		CVCL_0530
Experiment 6 Reporting the Activity Data of This PDC					[9]
Indication	Tumor
Efficacy Data	Cell viability	5%
Administration Time	72 h
Administration Dosage	10 µM
Description	The 72 h treatment is more effective than the 6 h pulse treatment. Notably, in vitro antitumor activities of 8 were found to correlate very good with the hY1R expression levels, as detected by gene expression analyses using RT-qPCR. Both the cytotoxic activity and the hY1R expression level rank in the order SK-N-MC > MDA-MB-468 > MDA-MB-231 > 184B5, what proofs the hY1R-specific and -selective nature of the mode of antitumor action of the designed PDC 8. Importantly, the activity of 8 against the selected normal breast cell line 184B5 is in the same order of magnitude as for the hY1R-deficient tumor cell line (MDA-MB-231), both tested at even higher concentration of the PDC than for the Y1 cell lines.    Click to Show/Hide
In Vitro Model	Breast adenocarcinoma	MDA-MB-468 cell		CVCL_0419
Experiment 7 Reporting the Activity Data of This PDC					[9]
Indication	Tumor
Efficacy Data	Cell viability	10%
Administration Time	6 h
Administration Dosage	10 µM
Description	The 72 h treatment is more effective than the 6 h pulse treatment. Notably, in vitro antitumor activities of 8 were found to correlate very good with the hY1R expression levels, as detected by gene expression analyses using RT-qPCR. Both the cytotoxic activity and the hY1R expression level rank in the order SK-N-MC > MDA-MB-468 > MDA-MB-231 > 184B5, what proofs the hY1R-specific and -selective nature of the mode of antitumor action of the designed PDC 8. Importantly, the activity of 8 against the selected normal breast cell line 184B5 is in the same order of magnitude as for the hY1R-deficient tumor cell line (MDA-MB-231), both tested at even higher concentration of the PDC than for the Y1 cell lines.    Click to Show/Hide
In Vitro Model	Breast adenocarcinoma	MDA-MB-468 cell		CVCL_0419
Experiment 8 Reporting the Activity Data of This PDC					[9]
Indication	Tumor
Efficacy Data	Cell viability	20%
Administration Time	72 h
Administration Dosage	10 µM
Description	The 72 h treatment is more effective than the 6 h pulse treatment. Notably, in vitro antitumor activities of 8 were found to correlate very good with the hY1R expression levels, as detected by gene expression analyses using RT-qPCR. Both the cytotoxic activity and the hY1R expression level rank in the order SK-N-MC > MDA-MB-468 > MDA-MB-231 > 184B5, what proofs the hY1R-specific and -selective nature of the mode of antitumor action of the designed PDC 8. Importantly, the activity of 8 against the selected normal breast cell line 184B5 is in the same order of magnitude as for the hY1R-deficient tumor cell line (MDA-MB-231), both tested at even higher concentration of the PDC than for the Y1 cell lines.    Click to Show/Hide
In Vitro Model	Breast adenocarcinoma	MDA-MB-231 cell		CVCL_0062
Experiment 9 Reporting the Activity Data of This PDC					[9]
Indication	Tumor
Efficacy Data	Cell viability	20%
Administration Time	72 h
Administration Dosage	10 µM
Description	The 72 h treatment is more effective than the 6 h pulse treatment. Notably, in vitro antitumor activities of 8 were found to correlate very good with the hY1R expression levels, as detected by gene expression analyses using RT-qPCR. Both the cytotoxic activity and the hY1R expression level rank in the order SK-N-MC > MDA-MB-468 > MDA-MB-231 > 184B5, what proofs the hY1R-specific and -selective nature of the mode of antitumor action of the designed PDC 8. Importantly, the activity of 8 against the selected normal breast cell line 184B5 is in the same order of magnitude as for the hY1R-deficient tumor cell line (MDA-MB-231), both tested at even higher concentration of the PDC than for the Y1 cell lines.    Click to Show/Hide
In Vitro Model	Normal	Normal mammary gland epithelium		Homo sapiens
Experiment 10 Reporting the Activity Data of This PDC					[9]
Indication	Tumor
Efficacy Data	Cell viability	35%
Administration Time	6 h
Administration Dosage	10 µM
Description	The 72 h treatment is more effective than the 6 h pulse treatment. Notably, in vitro antitumor activities of 8 were found to correlate very good with the hY1R expression levels, as detected by gene expression analyses using RT-qPCR. Both the cytotoxic activity and the hY1R expression level rank in the order SK-N-MC > MDA-MB-468 > MDA-MB-231 > 184B5, what proofs the hY1R-specific and -selective nature of the mode of antitumor action of the designed PDC 8. Importantly, the activity of 8 against the selected normal breast cell line 184B5 is in the same order of magnitude as for the hY1R-deficient tumor cell line (MDA-MB-231), both tested at even higher concentration of the PDC than for the Y1 cell lines.    Click to Show/Hide
In Vitro Model	Breast adenocarcinoma	MDA-MB-231 cell		CVCL_0062
Experiment 11 Reporting the Activity Data of This PDC					[9]
Indication	Tumor
Efficacy Data	Cell viability	50%
Administration Time	6 h
Administration Dosage	10 µM
Description	The 72 h treatment is more effective than the 6 h pulse treatment. Notably, in vitro antitumor activities of 8 were found to correlate very good with the hY1R expression levels, as detected by gene expression analyses using RT-qPCR. Both the cytotoxic activity and the hY1R expression level rank in the order SK-N-MC > MDA-MB-468 > MDA-MB-231 > 184B5, what proofs the hY1R-specific and -selective nature of the mode of antitumor action of the designed PDC 8. Importantly, the activity of 8 against the selected normal breast cell line 184B5 is in the same order of magnitude as for the hY1R-deficient tumor cell line (MDA-MB-231), both tested at even higher concentration of the PDC than for the Y1 cell lines.    Click to Show/Hide
In Vitro Model	Normal	Normal mammary gland epithelium		Homo sapiens

Experiment 1 Reporting the Activity Data of This PDC					[10]
Indication	Breast cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	236.1 nM
In Vitro Model	Breast adenocarcinoma	MDA-MB-468 cell		CVCL_0419
Experiment 2 Reporting the Activity Data of This PDC					[10]
Indication	Breast cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	405.3 nM
In Vitro Model	Invasive breast carcinoma	MCF-7 cell		CVCL_0031

Experiment 1 Reporting the Activity Data of This PDC					[11]
Indication	Colorectal cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	0.047 µM
Administration Time	48 h
Evaluation Method	MTT assay
MOA of PDC	Given these findings, we employed tissue-specific drug delivery approaches PDC to overcome those adverse effects. Several TfR-targeted peptides sequence with different affinity have been reported, so we synthesized a series of TfR-targeted peptide-DM1 conjugates for screening the effects of TfR-targeted drug candidates. It is speculated that the conjugates can specifically target tumor with high expression of TfR and can be uptake more than that of the normal cells. We succeeded found LWJ-M30, a conjugate of DM1 and B6, had a significant therapeutic effect after preliminary screening. In order to further reveal the improved therapeutic effects and mechanism of the LWJ-M30 on cancer with high TfR expression, in this study, we investigated the role of the LWJ-M30 in the targeted therapy for colorectal cancer in vitro and in vivo and explored the therapeutic mechanism.    Click to Show/Hide
Description	To determine the activity of the compounds, we analyzed the effects of them on cell growth and migration in HCT116 cells. We found that LWJ-M30 displayed significant inhibition of cell proliferation and clone formation ability. The IC50 of LWJ-M30 was 0.047 uM which was close to DM1 (0.026 uM). According to the results of wound-healing assay and transwell migration assay, LWJ-M30 also dramatically showed a inhibitory effect on cells migration. These results suggested that LWJ-M30 could inhibit the cells proliferation and migration.    Click to Show/Hide
In Vitro Model	Colon carcinoma	HCT 116 cell		CVCL_0291

Experiment 1 Reporting the Activity Data of This PDC					[6]
Indication	Breast cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	0.41 ± 0.13 µM
Administration Time	48 h
Evaluation Method	CCK-8 assay
MOA of PDC	In the previous study, we focused on the discovery of novel cell targeting peptides, leading to the identification of the preferred HER2-targeting peptide P1 (NPNWGRSWYNQRFK) derived from pertuzumab, with the Kd of 5.70 10-6 M. Here, we designed and synthesized a series of PDCs choosing Camptothecin (CPT) as the payload and P1 as the peptide carrier. We aimed to discover novel PDC molecules with potent inhibitory activity and selectivity targeting HER2 for effective cancer treatment. Conjugate Z8, which balanced the activity and the biosafety, showed good antiproliferative activity (IC50 = 1.91 ± 0.71 M), comparable to that of CPT (IC50 = 2.10 ± 1.34 M), against NCI-N87 cells. Moreover, subsequent verification tests indicated that Z8 operated through a mechanism similar to CPT, but the results from in vivo anti-tumor activity assay suggested that Z8 exhibited greater safety compared to CPT to some extent. These findings highlight Z8 as a promising candidate for further investigation in the treatment of HER2-positive tumors.    Click to Show/Hide
Description	To evaluated the in vitro anti-tumor activity of the conjugates, HER2-positive SK-BR-3, NCI-N87, HER2-negative MDA-MB-231 and normal MCF-10A cells were selected. As shown in Table 2, nearly all conjugates exhibited obvious anti-proliferation activity against SK-BR-3 cells, while most of the conjugates, except for Z6 and Z9, demonstrated different degrees of inhibitory activity on NCI-N87 cells. Taken together, Z8 and Z11 exhibited superior anti-tumor activity. The antiproliferative activity of Z8 against NCI-N87 cells (IC50 = 1.91 ± 0.71 uM) was comparable to that of CPT (IC50 = 2.10 ± 1.34 uM), while the antiproliferative activity of Z11 against SK-BR-3 cells (IC50 = 0.41 ± 0.13 uM) was comparable to that of CPT (IC50 = 0.26 ± 0.06 uM). Additionally, the IC50 values of these two conjugates against MDA-MB-231 and MCF-10A cells were significantly increased, showing their ability to selectively inhibit tumor cells. Based on the above results, Z8 and Z11, which exhibited better activity and apparent selectivity, were selected for subsequent studies.    Click to Show/Hide
In Vitro Model	Breast adenocarcinoma	SK-BR-3 cell		CVCL_0033
Experiment 2 Reporting the Activity Data of This PDC					[6]
Indication	Gastric cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	3.93 ± 1.41 µM
Administration Time	48 h
Evaluation Method	CCK-8 assay
MOA of PDC	In the previous study, we focused on the discovery of novel cell targeting peptides, leading to the identification of the preferred HER2-targeting peptide P1 (NPNWGRSWYNQRFK) derived from pertuzumab, with the Kd of 5.70 10-6 M. Here, we designed and synthesized a series of PDCs choosing Camptothecin (CPT) as the payload and P1 as the peptide carrier. We aimed to discover novel PDC molecules with potent inhibitory activity and selectivity targeting HER2 for effective cancer treatment. Conjugate Z8, which balanced the activity and the biosafety, showed good antiproliferative activity (IC50 = 1.91 ± 0.71 M), comparable to that of CPT (IC50 = 2.10 ± 1.34 M), against NCI-N87 cells. Moreover, subsequent verification tests indicated that Z8 operated through a mechanism similar to CPT, but the results from in vivo anti-tumor activity assay suggested that Z8 exhibited greater safety compared to CPT to some extent. These findings highlight Z8 as a promising candidate for further investigation in the treatment of HER2-positive tumors.    Click to Show/Hide
Description	To evaluated the in vitro anti-tumor activity of the conjugates, HER2-positive SK-BR-3, NCI-N87, HER2-negative MDA-MB-231 and normal MCF-10A cells were selected. As shown in Table 2, nearly all conjugates exhibited obvious anti-proliferation activity against SK-BR-3 cells, while most of the conjugates, except for Z6 and Z9, demonstrated different degrees of inhibitory activity on NCI-N87 cells. Taken together, Z8 and Z11 exhibited superior anti-tumor activity. The antiproliferative activity of Z8 against NCI-N87 cells (IC50 = 1.91 ± 0.71 uM) was comparable to that of CPT (IC50 = 2.10 ± 1.34 uM), while the antiproliferative activity of Z11 against SK-BR-3 cells (IC50 = 0.41 ± 0.13 uM) was comparable to that of CPT (IC50 = 0.26 ± 0.06 uM). Additionally, the IC50 values of these two conjugates against MDA-MB-231 and MCF-10A cells were significantly increased, showing their ability to selectively inhibit tumor cells. Based on the above results, Z8 and Z11, which exhibited better activity and apparent selectivity, were selected for subsequent studies.    Click to Show/Hide
In Vitro Model	Gastric tubular adenocarcinoma	NCI-N87 cell		CVCL_1603
Experiment 3 Reporting the Activity Data of This PDC					[6]
Indication	Breast cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	10.25 ± 1.34 µM
Administration Time	48 h
Evaluation Method	CCK-8 assay
MOA of PDC	In the previous study, we focused on the discovery of novel cell targeting peptides, leading to the identification of the preferred HER2-targeting peptide P1 (NPNWGRSWYNQRFK) derived from pertuzumab, with the Kd of 5.70 10-6 M. Here, we designed and synthesized a series of PDCs choosing Camptothecin (CPT) as the payload and P1 as the peptide carrier. We aimed to discover novel PDC molecules with potent inhibitory activity and selectivity targeting HER2 for effective cancer treatment. Conjugate Z8, which balanced the activity and the biosafety, showed good antiproliferative activity (IC50 = 1.91 ± 0.71 M), comparable to that of CPT (IC50 = 2.10 ± 1.34 M), against NCI-N87 cells. Moreover, subsequent verification tests indicated that Z8 operated through a mechanism similar to CPT, but the results from in vivo anti-tumor activity assay suggested that Z8 exhibited greater safety compared to CPT to some extent. These findings highlight Z8 as a promising candidate for further investigation in the treatment of HER2-positive tumors.    Click to Show/Hide
Description	To evaluated the in vitro anti-tumor activity of the conjugates, HER2-positive SK-BR-3, NCI-N87, HER2-negative MDA-MB-231 and normal MCF-10A cells were selected. As shown in Table 2, nearly all conjugates exhibited obvious anti-proliferation activity against SK-BR-3 cells, while most of the conjugates, except for Z6 and Z9, demonstrated different degrees of inhibitory activity on NCI-N87 cells. Taken together, Z8 and Z11 exhibited superior anti-tumor activity. The antiproliferative activity of Z8 against NCI-N87 cells (IC50 = 1.91 ± 0.71 uM) was comparable to that of CPT (IC50 = 2.10 ± 1.34 uM), while the antiproliferative activity of Z11 against SK-BR-3 cells (IC50 = 0.41 ± 0.13 uM) was comparable to that of CPT (IC50 = 0.26 ± 0.06 uM). Additionally, the IC50 values of these two conjugates against MDA-MB-231 and MCF-10A cells were significantly increased, showing their ability to selectively inhibit tumor cells. Based on the above results, Z8 and Z11, which exhibited better activity and apparent selectivity, were selected for subsequent studies.    Click to Show/Hide
In Vitro Model	Breast adenocarcinoma	MDA-MB-231 cell		CVCL_0062
Experiment 4 Reporting the Activity Data of This PDC					[6]
Indication	Breast cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	12.93 ± 1.15 µM
Administration Time	48 h
Evaluation Method	CCK-8 assay
MOA of PDC	In the previous study, we focused on the discovery of novel cell targeting peptides, leading to the identification of the preferred HER2-targeting peptide P1 (NPNWGRSWYNQRFK) derived from pertuzumab, with the Kd of 5.70 10-6 M. Here, we designed and synthesized a series of PDCs choosing Camptothecin (CPT) as the payload and P1 as the peptide carrier. We aimed to discover novel PDC molecules with potent inhibitory activity and selectivity targeting HER2 for effective cancer treatment. Conjugate Z8, which balanced the activity and the biosafety, showed good antiproliferative activity (IC50 = 1.91 ± 0.71 M), comparable to that of CPT (IC50 = 2.10 ± 1.34 M), against NCI-N87 cells. Moreover, subsequent verification tests indicated that Z8 operated through a mechanism similar to CPT, but the results from in vivo anti-tumor activity assay suggested that Z8 exhibited greater safety compared to CPT to some extent. These findings highlight Z8 as a promising candidate for further investigation in the treatment of HER2-positive tumors.    Click to Show/Hide
Description	To evaluated the in vitro anti-tumor activity of the conjugates, HER2-positive SK-BR-3, NCI-N87, HER2-negative MDA-MB-231 and normal MCF-10A cells were selected. As shown in Table 2, nearly all conjugates exhibited obvious anti-proliferation activity against SK-BR-3 cells, while most of the conjugates, except for Z6 and Z9, demonstrated different degrees of inhibitory activity on NCI-N87 cells. Taken together, Z8 and Z11 exhibited superior anti-tumor activity. The antiproliferative activity of Z8 against NCI-N87 cells (IC50 = 1.91 ± 0.71 uM) was comparable to that of CPT (IC50 = 2.10 ± 1.34 uM), while the antiproliferative activity of Z11 against SK-BR-3 cells (IC50 = 0.41 ± 0.13 uM) was comparable to that of CPT (IC50 = 0.26 ± 0.06 uM). Additionally, the IC50 values of these two conjugates against MDA-MB-231 and MCF-10A cells were significantly increased, showing their ability to selectively inhibit tumor cells. Based on the above results, Z8 and Z11, which exhibited better activity and apparent selectivity, were selected for subsequent studies.    Click to Show/Hide
In Vitro Model	Normal	MCF-10A cell		CVCL_0598

Experiment 1 Reporting the Activity Data of This PDC					[12]
Indication	Hepatocellular carcinoma
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	0.87 µM
In Vitro Model	Hepatocellular carcinoma	SMMC-7721 cell		CVCL_0534
Experiment 2 Reporting the Activity Data of This PDC					[12]
Indication	Breast cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	3.66 µM
In Vitro Model	Invasive breast carcinoma of no special type	EGFR-overexpressing MCF-7 cell		CVCL_0031
Experiment 3 Reporting the Activity Data of This PDC					[12]
Indication	Breast cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	32.2µM
In Vitro Model	Invasive breast carcinoma	MCF-7 cell		CVCL_0031

Experiment 1 Reporting the Activity Data of This PDC					[6]
Indication	Breast cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	1.37 ± 0.42 µM
Administration Time	48 h
Evaluation Method	CCK-8 assay
MOA of PDC	In the previous study, we focused on the discovery of novel cell targeting peptides, leading to the identification of the preferred HER2-targeting peptide P1 (NPNWGRSWYNQRFK) derived from pertuzumab, with the Kd of 5.70 10-6 M. Here, we designed and synthesized a series of PDCs choosing Camptothecin (CPT) as the payload and P1 as the peptide carrier. We aimed to discover novel PDC molecules with potent inhibitory activity and selectivity targeting HER2 for effective cancer treatment. Conjugate Z8, which balanced the activity and the biosafety, showed good antiproliferative activity (IC50 = 1.91 ± 0.71 M), comparable to that of CPT (IC50 = 2.10 ± 1.34 M), against NCI-N87 cells. Moreover, subsequent verification tests indicated that Z8 operated through a mechanism similar to CPT, but the results from in vivo anti-tumor activity assay suggested that Z8 exhibited greater safety compared to CPT to some extent. These findings highlight Z8 as a promising candidate for further investigation in the treatment of HER2-positive tumors.    Click to Show/Hide
Description	To evaluated the in vitro anti-tumor activity of the conjugates, HER2-positive SK-BR-3, NCI-N87, HER2-negative MDA-MB-231 and normal MCF-10A cells were selected. As shown in Table 2, nearly all conjugates exhibited obvious anti-proliferation activity against SK-BR-3 cells, while most of the conjugates, except for Z6 and Z9, demonstrated different degrees of inhibitory activity on NCI-N87 cells. Taken together, Z8 and Z11 exhibited superior anti-tumor activity. The antiproliferative activity of Z8 against NCI-N87 cells (IC50 = 1.91 ± 0.71 uM) was comparable to that of CPT (IC50 = 2.10 ± 1.34 uM), while the antiproliferative activity of Z11 against SK-BR-3 cells (IC50 = 0.41 ± 0.13 uM) was comparable to that of CPT (IC50 = 0.26 ± 0.06 uM). Additionally, the IC50 values of these two conjugates against MDA-MB-231 and MCF-10A cells were significantly increased, showing their ability to selectively inhibit tumor cells. Based on the above results, Z8 and Z11, which exhibited better activity and apparent selectivity, were selected for subsequent studies.    Click to Show/Hide
In Vitro Model	Breast adenocarcinoma	SK-BR-3 cell		CVCL_0033
Experiment 2 Reporting the Activity Data of This PDC					[6]
Indication	Gastric cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	3.45 ± 1.09 µM
Administration Time	48 h
Evaluation Method	CCK-8 assay
MOA of PDC	In the previous study, we focused on the discovery of novel cell targeting peptides, leading to the identification of the preferred HER2-targeting peptide P1 (NPNWGRSWYNQRFK) derived from pertuzumab, with the Kd of 5.70 10-6 M. Here, we designed and synthesized a series of PDCs choosing Camptothecin (CPT) as the payload and P1 as the peptide carrier. We aimed to discover novel PDC molecules with potent inhibitory activity and selectivity targeting HER2 for effective cancer treatment. Conjugate Z8, which balanced the activity and the biosafety, showed good antiproliferative activity (IC50 = 1.91 ± 0.71 M), comparable to that of CPT (IC50 = 2.10 ± 1.34 M), against NCI-N87 cells. Moreover, subsequent verification tests indicated that Z8 operated through a mechanism similar to CPT, but the results from in vivo anti-tumor activity assay suggested that Z8 exhibited greater safety compared to CPT to some extent. These findings highlight Z8 as a promising candidate for further investigation in the treatment of HER2-positive tumors.    Click to Show/Hide
Description	To evaluated the in vitro anti-tumor activity of the conjugates, HER2-positive SK-BR-3, NCI-N87, HER2-negative MDA-MB-231 and normal MCF-10A cells were selected. As shown in Table 2, nearly all conjugates exhibited obvious anti-proliferation activity against SK-BR-3 cells, while most of the conjugates, except for Z6 and Z9, demonstrated different degrees of inhibitory activity on NCI-N87 cells. Taken together, Z8 and Z11 exhibited superior anti-tumor activity. The antiproliferative activity of Z8 against NCI-N87 cells (IC50 = 1.91 ± 0.71 uM) was comparable to that of CPT (IC50 = 2.10 ± 1.34 uM), while the antiproliferative activity of Z11 against SK-BR-3 cells (IC50 = 0.41 ± 0.13 uM) was comparable to that of CPT (IC50 = 0.26 ± 0.06 uM). Additionally, the IC50 values of these two conjugates against MDA-MB-231 and MCF-10A cells were significantly increased, showing their ability to selectively inhibit tumor cells. Based on the above results, Z8 and Z11, which exhibited better activity and apparent selectivity, were selected for subsequent studies.    Click to Show/Hide
In Vitro Model	Gastric tubular adenocarcinoma	NCI-N87 cell		CVCL_1603
Experiment 3 Reporting the Activity Data of This PDC					[6]
Indication	Breast cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	13.91 ± 2.61 µM
Administration Time	48 h
Evaluation Method	CCK-8 assay
MOA of PDC	In the previous study, we focused on the discovery of novel cell targeting peptides, leading to the identification of the preferred HER2-targeting peptide P1 (NPNWGRSWYNQRFK) derived from pertuzumab, with the Kd of 5.70 10-6 M. Here, we designed and synthesized a series of PDCs choosing Camptothecin (CPT) as the payload and P1 as the peptide carrier. We aimed to discover novel PDC molecules with potent inhibitory activity and selectivity targeting HER2 for effective cancer treatment. Conjugate Z8, which balanced the activity and the biosafety, showed good antiproliferative activity (IC50 = 1.91 ± 0.71 M), comparable to that of CPT (IC50 = 2.10 ± 1.34 M), against NCI-N87 cells. Moreover, subsequent verification tests indicated that Z8 operated through a mechanism similar to CPT, but the results from in vivo anti-tumor activity assay suggested that Z8 exhibited greater safety compared to CPT to some extent. These findings highlight Z8 as a promising candidate for further investigation in the treatment of HER2-positive tumors.    Click to Show/Hide
Description	To evaluated the in vitro anti-tumor activity of the conjugates, HER2-positive SK-BR-3, NCI-N87, HER2-negative MDA-MB-231 and normal MCF-10A cells were selected. As shown in Table 2, nearly all conjugates exhibited obvious anti-proliferation activity against SK-BR-3 cells, while most of the conjugates, except for Z6 and Z9, demonstrated different degrees of inhibitory activity on NCI-N87 cells. Taken together, Z8 and Z11 exhibited superior anti-tumor activity. The antiproliferative activity of Z8 against NCI-N87 cells (IC50 = 1.91 ± 0.71 uM) was comparable to that of CPT (IC50 = 2.10 ± 1.34 uM), while the antiproliferative activity of Z11 against SK-BR-3 cells (IC50 = 0.41 ± 0.13 uM) was comparable to that of CPT (IC50 = 0.26 ± 0.06 uM). Additionally, the IC50 values of these two conjugates against MDA-MB-231 and MCF-10A cells were significantly increased, showing their ability to selectively inhibit tumor cells. Based on the above results, Z8 and Z11, which exhibited better activity and apparent selectivity, were selected for subsequent studies.    Click to Show/Hide
In Vitro Model	Breast adenocarcinoma	MDA-MB-231 cell		CVCL_0062
Experiment 4 Reporting the Activity Data of This PDC					[6]
Indication	Breast cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	28.25 ± 3.12 µM
Administration Time	48 h
Evaluation Method	CCK-8 assay
MOA of PDC	In the previous study, we focused on the discovery of novel cell targeting peptides, leading to the identification of the preferred HER2-targeting peptide P1 (NPNWGRSWYNQRFK) derived from pertuzumab, with the Kd of 5.70 10-6 M. Here, we designed and synthesized a series of PDCs choosing Camptothecin (CPT) as the payload and P1 as the peptide carrier. We aimed to discover novel PDC molecules with potent inhibitory activity and selectivity targeting HER2 for effective cancer treatment. Conjugate Z8, which balanced the activity and the biosafety, showed good antiproliferative activity (IC50 = 1.91 ± 0.71 M), comparable to that of CPT (IC50 = 2.10 ± 1.34 M), against NCI-N87 cells. Moreover, subsequent verification tests indicated that Z8 operated through a mechanism similar to CPT, but the results from in vivo anti-tumor activity assay suggested that Z8 exhibited greater safety compared to CPT to some extent. These findings highlight Z8 as a promising candidate for further investigation in the treatment of HER2-positive tumors.    Click to Show/Hide
Description	To evaluated the in vitro anti-tumor activity of the conjugates, HER2-positive SK-BR-3, NCI-N87, HER2-negative MDA-MB-231 and normal MCF-10A cells were selected. As shown in Table 2, nearly all conjugates exhibited obvious anti-proliferation activity against SK-BR-3 cells, while most of the conjugates, except for Z6 and Z9, demonstrated different degrees of inhibitory activity on NCI-N87 cells. Taken together, Z8 and Z11 exhibited superior anti-tumor activity. The antiproliferative activity of Z8 against NCI-N87 cells (IC50 = 1.91 ± 0.71 uM) was comparable to that of CPT (IC50 = 2.10 ± 1.34 uM), while the antiproliferative activity of Z11 against SK-BR-3 cells (IC50 = 0.41 ± 0.13 uM) was comparable to that of CPT (IC50 = 0.26 ± 0.06 uM). Additionally, the IC50 values of these two conjugates against MDA-MB-231 and MCF-10A cells were significantly increased, showing their ability to selectively inhibit tumor cells. Based on the above results, Z8 and Z11, which exhibited better activity and apparent selectivity, were selected for subsequent studies.    Click to Show/Hide
In Vitro Model	Normal	MCF-10A cell		CVCL_0598

Experiment 1 Reporting the Activity Data of This PDC					[6]
Indication	Breast cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	1.44 ± 0.63 µM
Administration Time	48 h
Evaluation Method	CCK-8 assay
MOA of PDC	In the previous study, we focused on the discovery of novel cell targeting peptides, leading to the identification of the preferred HER2-targeting peptide P1 (NPNWGRSWYNQRFK) derived from pertuzumab, with the Kd of 5.70 10-6 M. Here, we designed and synthesized a series of PDCs choosing Camptothecin (CPT) as the payload and P1 as the peptide carrier. We aimed to discover novel PDC molecules with potent inhibitory activity and selectivity targeting HER2 for effective cancer treatment. Conjugate Z8, which balanced the activity and the biosafety, showed good antiproliferative activity (IC50 = 1.91 ± 0.71 M), comparable to that of CPT (IC50 = 2.10 ± 1.34 M), against NCI-N87 cells. Moreover, subsequent verification tests indicated that Z8 operated through a mechanism similar to CPT, but the results from in vivo anti-tumor activity assay suggested that Z8 exhibited greater safety compared to CPT to some extent. These findings highlight Z8 as a promising candidate for further investigation in the treatment of HER2-positive tumors.    Click to Show/Hide
Description	To evaluated the in vitro anti-tumor activity of the conjugates, HER2-positive SK-BR-3, NCI-N87, HER2-negative MDA-MB-231 and normal MCF-10A cells were selected. As shown in Table 2, nearly all conjugates exhibited obvious anti-proliferation activity against SK-BR-3 cells, while most of the conjugates, except for Z6 and Z9, demonstrated different degrees of inhibitory activity on NCI-N87 cells. Taken together, Z8 and Z11 exhibited superior anti-tumor activity. The antiproliferative activity of Z8 against NCI-N87 cells (IC50 = 1.91 ± 0.71 uM) was comparable to that of CPT (IC50 = 2.10 ± 1.34 uM), while the antiproliferative activity of Z11 against SK-BR-3 cells (IC50 = 0.41 ± 0.13 uM) was comparable to that of CPT (IC50 = 0.26 ± 0.06 uM). Additionally, the IC50 values of these two conjugates against MDA-MB-231 and MCF-10A cells were significantly increased, showing their ability to selectively inhibit tumor cells. Based on the above results, Z8 and Z11, which exhibited better activity and apparent selectivity, were selected for subsequent studies.    Click to Show/Hide
In Vitro Model	Breast adenocarcinoma	SK-BR-3 cell		CVCL_0033
Experiment 2 Reporting the Activity Data of This PDC					[6]
Indication	Gastric cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	7.83 ± 1.77 µM
Administration Time	48 h
Evaluation Method	CCK-8 assay
MOA of PDC	In the previous study, we focused on the discovery of novel cell targeting peptides, leading to the identification of the preferred HER2-targeting peptide P1 (NPNWGRSWYNQRFK) derived from pertuzumab, with the Kd of 5.70 10-6 M. Here, we designed and synthesized a series of PDCs choosing Camptothecin (CPT) as the payload and P1 as the peptide carrier. We aimed to discover novel PDC molecules with potent inhibitory activity and selectivity targeting HER2 for effective cancer treatment. Conjugate Z8, which balanced the activity and the biosafety, showed good antiproliferative activity (IC50 = 1.91 ± 0.71 M), comparable to that of CPT (IC50 = 2.10 ± 1.34 M), against NCI-N87 cells. Moreover, subsequent verification tests indicated that Z8 operated through a mechanism similar to CPT, but the results from in vivo anti-tumor activity assay suggested that Z8 exhibited greater safety compared to CPT to some extent. These findings highlight Z8 as a promising candidate for further investigation in the treatment of HER2-positive tumors.    Click to Show/Hide
Description	To evaluated the in vitro anti-tumor activity of the conjugates, HER2-positive SK-BR-3, NCI-N87, HER2-negative MDA-MB-231 and normal MCF-10A cells were selected. As shown in Table 2, nearly all conjugates exhibited obvious anti-proliferation activity against SK-BR-3 cells, while most of the conjugates, except for Z6 and Z9, demonstrated different degrees of inhibitory activity on NCI-N87 cells. Taken together, Z8 and Z11 exhibited superior anti-tumor activity. The antiproliferative activity of Z8 against NCI-N87 cells (IC50 = 1.91 ± 0.71 uM) was comparable to that of CPT (IC50 = 2.10 ± 1.34 uM), while the antiproliferative activity of Z11 against SK-BR-3 cells (IC50 = 0.41 ± 0.13 uM) was comparable to that of CPT (IC50 = 0.26 ± 0.06 uM). Additionally, the IC50 values of these two conjugates against MDA-MB-231 and MCF-10A cells were significantly increased, showing their ability to selectively inhibit tumor cells. Based on the above results, Z8 and Z11, which exhibited better activity and apparent selectivity, were selected for subsequent studies.    Click to Show/Hide
In Vitro Model	Gastric tubular adenocarcinoma	NCI-N87 cell		CVCL_1603
Experiment 3 Reporting the Activity Data of This PDC					[6]
Indication	Breast cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	17.84 ± 3.14 µM
Administration Time	48 h
Evaluation Method	CCK-8 assay
MOA of PDC	In the previous study, we focused on the discovery of novel cell targeting peptides, leading to the identification of the preferred HER2-targeting peptide P1 (NPNWGRSWYNQRFK) derived from pertuzumab, with the Kd of 5.70 10-6 M. Here, we designed and synthesized a series of PDCs choosing Camptothecin (CPT) as the payload and P1 as the peptide carrier. We aimed to discover novel PDC molecules with potent inhibitory activity and selectivity targeting HER2 for effective cancer treatment. Conjugate Z8, which balanced the activity and the biosafety, showed good antiproliferative activity (IC50 = 1.91 ± 0.71 M), comparable to that of CPT (IC50 = 2.10 ± 1.34 M), against NCI-N87 cells. Moreover, subsequent verification tests indicated that Z8 operated through a mechanism similar to CPT, but the results from in vivo anti-tumor activity assay suggested that Z8 exhibited greater safety compared to CPT to some extent. These findings highlight Z8 as a promising candidate for further investigation in the treatment of HER2-positive tumors.    Click to Show/Hide
Description	To evaluated the in vitro anti-tumor activity of the conjugates, HER2-positive SK-BR-3, NCI-N87, HER2-negative MDA-MB-231 and normal MCF-10A cells were selected. As shown in Table 2, nearly all conjugates exhibited obvious anti-proliferation activity against SK-BR-3 cells, while most of the conjugates, except for Z6 and Z9, demonstrated different degrees of inhibitory activity on NCI-N87 cells. Taken together, Z8 and Z11 exhibited superior anti-tumor activity. The antiproliferative activity of Z8 against NCI-N87 cells (IC50 = 1.91 ± 0.71 uM) was comparable to that of CPT (IC50 = 2.10 ± 1.34 uM), while the antiproliferative activity of Z11 against SK-BR-3 cells (IC50 = 0.41 ± 0.13 uM) was comparable to that of CPT (IC50 = 0.26 ± 0.06 uM). Additionally, the IC50 values of these two conjugates against MDA-MB-231 and MCF-10A cells were significantly increased, showing their ability to selectively inhibit tumor cells. Based on the above results, Z8 and Z11, which exhibited better activity and apparent selectivity, were selected for subsequent studies.    Click to Show/Hide
In Vitro Model	Breast adenocarcinoma	MDA-MB-231 cell		CVCL_0062
Experiment 4 Reporting the Activity Data of This PDC					[6]
Indication	Breast cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	20.36 ± 1.82 µM
Administration Time	48 h
Evaluation Method	CCK-8 assay
MOA of PDC	In the previous study, we focused on the discovery of novel cell targeting peptides, leading to the identification of the preferred HER2-targeting peptide P1 (NPNWGRSWYNQRFK) derived from pertuzumab, with the Kd of 5.70 10-6 M. Here, we designed and synthesized a series of PDCs choosing Camptothecin (CPT) as the payload and P1 as the peptide carrier. We aimed to discover novel PDC molecules with potent inhibitory activity and selectivity targeting HER2 for effective cancer treatment. Conjugate Z8, which balanced the activity and the biosafety, showed good antiproliferative activity (IC50 = 1.91 ± 0.71 M), comparable to that of CPT (IC50 = 2.10 ± 1.34 M), against NCI-N87 cells. Moreover, subsequent verification tests indicated that Z8 operated through a mechanism similar to CPT, but the results from in vivo anti-tumor activity assay suggested that Z8 exhibited greater safety compared to CPT to some extent. These findings highlight Z8 as a promising candidate for further investigation in the treatment of HER2-positive tumors.    Click to Show/Hide
Description	To evaluated the in vitro anti-tumor activity of the conjugates, HER2-positive SK-BR-3, NCI-N87, HER2-negative MDA-MB-231 and normal MCF-10A cells were selected. As shown in Table 2, nearly all conjugates exhibited obvious anti-proliferation activity against SK-BR-3 cells, while most of the conjugates, except for Z6 and Z9, demonstrated different degrees of inhibitory activity on NCI-N87 cells. Taken together, Z8 and Z11 exhibited superior anti-tumor activity. The antiproliferative activity of Z8 against NCI-N87 cells (IC50 = 1.91 ± 0.71 uM) was comparable to that of CPT (IC50 = 2.10 ± 1.34 uM), while the antiproliferative activity of Z11 against SK-BR-3 cells (IC50 = 0.41 ± 0.13 uM) was comparable to that of CPT (IC50 = 0.26 ± 0.06 uM). Additionally, the IC50 values of these two conjugates against MDA-MB-231 and MCF-10A cells were significantly increased, showing their ability to selectively inhibit tumor cells. Based on the above results, Z8 and Z11, which exhibited better activity and apparent selectivity, were selected for subsequent studies.    Click to Show/Hide
In Vitro Model	Normal	MCF-10A cell		CVCL_0598

Experiment 1 Reporting the Activity Data of This PDC					[6]
Indication	Breast cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	1.76 ± 0.33 µM
Administration Time	48 h
Evaluation Method	CCK-8 assay
MOA of PDC	In the previous study, we focused on the discovery of novel cell targeting peptides, leading to the identification of the preferred HER2-targeting peptide P1 (NPNWGRSWYNQRFK) derived from pertuzumab, with the Kd of 5.70 10-6 M. Here, we designed and synthesized a series of PDCs choosing Camptothecin (CPT) as the payload and P1 as the peptide carrier. We aimed to discover novel PDC molecules with potent inhibitory activity and selectivity targeting HER2 for effective cancer treatment. Conjugate Z8, which balanced the activity and the biosafety, showed good antiproliferative activity (IC50 = 1.91 ± 0.71 M), comparable to that of CPT (IC50 = 2.10 ± 1.34 M), against NCI-N87 cells. Moreover, subsequent verification tests indicated that Z8 operated through a mechanism similar to CPT, but the results from in vivo anti-tumor activity assay suggested that Z8 exhibited greater safety compared to CPT to some extent. These findings highlight Z8 as a promising candidate for further investigation in the treatment of HER2-positive tumors.    Click to Show/Hide
Description	To evaluated the in vitro anti-tumor activity of the conjugates, HER2-positive SK-BR-3, NCI-N87, HER2-negative MDA-MB-231 and normal MCF-10A cells were selected. As shown in Table 2, nearly all conjugates exhibited obvious anti-proliferation activity against SK-BR-3 cells, while most of the conjugates, except for Z6 and Z9, demonstrated different degrees of inhibitory activity on NCI-N87 cells. Taken together, Z8 and Z11 exhibited superior anti-tumor activity. The antiproliferative activity of Z8 against NCI-N87 cells (IC50 = 1.91 ± 0.71 uM) was comparable to that of CPT (IC50 = 2.10 ± 1.34 uM), while the antiproliferative activity of Z11 against SK-BR-3 cells (IC50 = 0.41 ± 0.13 uM) was comparable to that of CPT (IC50 = 0.26 ± 0.06 uM). Additionally, the IC50 values of these two conjugates against MDA-MB-231 and MCF-10A cells were significantly increased, showing their ability to selectively inhibit tumor cells. Based on the above results, Z8 and Z11, which exhibited better activity and apparent selectivity, were selected for subsequent studies.    Click to Show/Hide
In Vitro Model	Breast adenocarcinoma	SK-BR-3 cell		CVCL_0033
Experiment 2 Reporting the Activity Data of This PDC					[6]
Indication	Gastric cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	6.12 ± 0.45 µM
Administration Time	48 h
Evaluation Method	CCK-8 assay
MOA of PDC	In the previous study, we focused on the discovery of novel cell targeting peptides, leading to the identification of the preferred HER2-targeting peptide P1 (NPNWGRSWYNQRFK) derived from pertuzumab, with the Kd of 5.70 10-6 M. Here, we designed and synthesized a series of PDCs choosing Camptothecin (CPT) as the payload and P1 as the peptide carrier. We aimed to discover novel PDC molecules with potent inhibitory activity and selectivity targeting HER2 for effective cancer treatment. Conjugate Z8, which balanced the activity and the biosafety, showed good antiproliferative activity (IC50 = 1.91 ± 0.71 M), comparable to that of CPT (IC50 = 2.10 ± 1.34 M), against NCI-N87 cells. Moreover, subsequent verification tests indicated that Z8 operated through a mechanism similar to CPT, but the results from in vivo anti-tumor activity assay suggested that Z8 exhibited greater safety compared to CPT to some extent. These findings highlight Z8 as a promising candidate for further investigation in the treatment of HER2-positive tumors.    Click to Show/Hide
Description	To evaluated the in vitro anti-tumor activity of the conjugates, HER2-positive SK-BR-3, NCI-N87, HER2-negative MDA-MB-231 and normal MCF-10A cells were selected. As shown in Table 2, nearly all conjugates exhibited obvious anti-proliferation activity against SK-BR-3 cells, while most of the conjugates, except for Z6 and Z9, demonstrated different degrees of inhibitory activity on NCI-N87 cells. Taken together, Z8 and Z11 exhibited superior anti-tumor activity. The antiproliferative activity of Z8 against NCI-N87 cells (IC50 = 1.91 ± 0.71 uM) was comparable to that of CPT (IC50 = 2.10 ± 1.34 uM), while the antiproliferative activity of Z11 against SK-BR-3 cells (IC50 = 0.41 ± 0.13 uM) was comparable to that of CPT (IC50 = 0.26 ± 0.06 uM). Additionally, the IC50 values of these two conjugates against MDA-MB-231 and MCF-10A cells were significantly increased, showing their ability to selectively inhibit tumor cells. Based on the above results, Z8 and Z11, which exhibited better activity and apparent selectivity, were selected for subsequent studies.    Click to Show/Hide
In Vitro Model	Gastric tubular adenocarcinoma	NCI-N87 cell		CVCL_1603
Experiment 3 Reporting the Activity Data of This PDC					[6]
Indication	Breast cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	13.21 ± 1.15 µM
Administration Time	48 h
Evaluation Method	CCK-8 assay
MOA of PDC	In the previous study, we focused on the discovery of novel cell targeting peptides, leading to the identification of the preferred HER2-targeting peptide P1 (NPNWGRSWYNQRFK) derived from pertuzumab, with the Kd of 5.70 10-6 M. Here, we designed and synthesized a series of PDCs choosing Camptothecin (CPT) as the payload and P1 as the peptide carrier. We aimed to discover novel PDC molecules with potent inhibitory activity and selectivity targeting HER2 for effective cancer treatment. Conjugate Z8, which balanced the activity and the biosafety, showed good antiproliferative activity (IC50 = 1.91 ± 0.71 M), comparable to that of CPT (IC50 = 2.10 ± 1.34 M), against NCI-N87 cells. Moreover, subsequent verification tests indicated that Z8 operated through a mechanism similar to CPT, but the results from in vivo anti-tumor activity assay suggested that Z8 exhibited greater safety compared to CPT to some extent. These findings highlight Z8 as a promising candidate for further investigation in the treatment of HER2-positive tumors.    Click to Show/Hide
Description	To evaluated the in vitro anti-tumor activity of the conjugates, HER2-positive SK-BR-3, NCI-N87, HER2-negative MDA-MB-231 and normal MCF-10A cells were selected. As shown in Table 2, nearly all conjugates exhibited obvious anti-proliferation activity against SK-BR-3 cells, while most of the conjugates, except for Z6 and Z9, demonstrated different degrees of inhibitory activity on NCI-N87 cells. Taken together, Z8 and Z11 exhibited superior anti-tumor activity. The antiproliferative activity of Z8 against NCI-N87 cells (IC50 = 1.91 ± 0.71 uM) was comparable to that of CPT (IC50 = 2.10 ± 1.34 uM), while the antiproliferative activity of Z11 against SK-BR-3 cells (IC50 = 0.41 ± 0.13 uM) was comparable to that of CPT (IC50 = 0.26 ± 0.06 uM). Additionally, the IC50 values of these two conjugates against MDA-MB-231 and MCF-10A cells were significantly increased, showing their ability to selectively inhibit tumor cells. Based on the above results, Z8 and Z11, which exhibited better activity and apparent selectivity, were selected for subsequent studies.    Click to Show/Hide
In Vitro Model	Breast adenocarcinoma	MDA-MB-231 cell		CVCL_0062
Experiment 4 Reporting the Activity Data of This PDC					[6]
Indication	Breast cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	15.25 ± 2.08 µM
Administration Time	48 h
Evaluation Method	CCK-8 assay
MOA of PDC	In the previous study, we focused on the discovery of novel cell targeting peptides, leading to the identification of the preferred HER2-targeting peptide P1 (NPNWGRSWYNQRFK) derived from pertuzumab, with the Kd of 5.70 10-6 M. Here, we designed and synthesized a series of PDCs choosing Camptothecin (CPT) as the payload and P1 as the peptide carrier. We aimed to discover novel PDC molecules with potent inhibitory activity and selectivity targeting HER2 for effective cancer treatment. Conjugate Z8, which balanced the activity and the biosafety, showed good antiproliferative activity (IC50 = 1.91 ± 0.71 M), comparable to that of CPT (IC50 = 2.10 ± 1.34 M), against NCI-N87 cells. Moreover, subsequent verification tests indicated that Z8 operated through a mechanism similar to CPT, but the results from in vivo anti-tumor activity assay suggested that Z8 exhibited greater safety compared to CPT to some extent. These findings highlight Z8 as a promising candidate for further investigation in the treatment of HER2-positive tumors.    Click to Show/Hide
Description	To evaluated the in vitro anti-tumor activity of the conjugates, HER2-positive SK-BR-3, NCI-N87, HER2-negative MDA-MB-231 and normal MCF-10A cells were selected. As shown in Table 2, nearly all conjugates exhibited obvious anti-proliferation activity against SK-BR-3 cells, while most of the conjugates, except for Z6 and Z9, demonstrated different degrees of inhibitory activity on NCI-N87 cells. Taken together, Z8 and Z11 exhibited superior anti-tumor activity. The antiproliferative activity of Z8 against NCI-N87 cells (IC50 = 1.91 ± 0.71 uM) was comparable to that of CPT (IC50 = 2.10 ± 1.34 uM), while the antiproliferative activity of Z11 against SK-BR-3 cells (IC50 = 0.41 ± 0.13 uM) was comparable to that of CPT (IC50 = 0.26 ± 0.06 uM). Additionally, the IC50 values of these two conjugates against MDA-MB-231 and MCF-10A cells were significantly increased, showing their ability to selectively inhibit tumor cells. Based on the above results, Z8 and Z11, which exhibited better activity and apparent selectivity, were selected for subsequent studies.    Click to Show/Hide
In Vitro Model	Normal	MCF-10A cell		CVCL_0598

Experiment 1 Reporting the Activity Data of This PDC					[6]
Indication	Breast cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	2.37 ± 0.32 µM
Administration Time	48 h
Evaluation Method	CCK-8 assay
MOA of PDC	In the previous study, we focused on the discovery of novel cell targeting peptides, leading to the identification of the preferred HER2-targeting peptide P1 (NPNWGRSWYNQRFK) derived from pertuzumab, with the Kd of 5.70 10-6 M. Here, we designed and synthesized a series of PDCs choosing Camptothecin (CPT) as the payload and P1 as the peptide carrier. We aimed to discover novel PDC molecules with potent inhibitory activity and selectivity targeting HER2 for effective cancer treatment. Conjugate Z8, which balanced the activity and the biosafety, showed good antiproliferative activity (IC50 = 1.91 ± 0.71 M), comparable to that of CPT (IC50 = 2.10 ± 1.34 M), against NCI-N87 cells. Moreover, subsequent verification tests indicated that Z8 operated through a mechanism similar to CPT, but the results from in vivo anti-tumor activity assay suggested that Z8 exhibited greater safety compared to CPT to some extent. These findings highlight Z8 as a promising candidate for further investigation in the treatment of HER2-positive tumors.    Click to Show/Hide
Description	To evaluated the in vitro anti-tumor activity of the conjugates, HER2-positive SK-BR-3, NCI-N87, HER2-negative MDA-MB-231 and normal MCF-10A cells were selected. As shown in Table 2, nearly all conjugates exhibited obvious anti-proliferation activity against SK-BR-3 cells, while most of the conjugates, except for Z6 and Z9, demonstrated different degrees of inhibitory activity on NCI-N87 cells. Taken together, Z8 and Z11 exhibited superior anti-tumor activity. The antiproliferative activity of Z8 against NCI-N87 cells (IC50 = 1.91 ± 0.71 uM) was comparable to that of CPT (IC50 = 2.10 ± 1.34 uM), while the antiproliferative activity of Z11 against SK-BR-3 cells (IC50 = 0.41 ± 0.13 uM) was comparable to that of CPT (IC50 = 0.26 ± 0.06 uM). Additionally, the IC50 values of these two conjugates against MDA-MB-231 and MCF-10A cells were significantly increased, showing their ability to selectively inhibit tumor cells. Based on the above results, Z8 and Z11, which exhibited better activity and apparent selectivity, were selected for subsequent studies.    Click to Show/Hide
In Vitro Model	Breast adenocarcinoma	SK-BR-3 cell		CVCL_0033
Experiment 2 Reporting the Activity Data of This PDC					[6]
Indication	Gastric cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	7.25 ± 1.75 µM
Administration Time	48 h
Evaluation Method	CCK-8 assay
MOA of PDC	In the previous study, we focused on the discovery of novel cell targeting peptides, leading to the identification of the preferred HER2-targeting peptide P1 (NPNWGRSWYNQRFK) derived from pertuzumab, with the Kd of 5.70 10-6 M. Here, we designed and synthesized a series of PDCs choosing Camptothecin (CPT) as the payload and P1 as the peptide carrier. We aimed to discover novel PDC molecules with potent inhibitory activity and selectivity targeting HER2 for effective cancer treatment. Conjugate Z8, which balanced the activity and the biosafety, showed good antiproliferative activity (IC50 = 1.91 ± 0.71 M), comparable to that of CPT (IC50 = 2.10 ± 1.34 M), against NCI-N87 cells. Moreover, subsequent verification tests indicated that Z8 operated through a mechanism similar to CPT, but the results from in vivo anti-tumor activity assay suggested that Z8 exhibited greater safety compared to CPT to some extent. These findings highlight Z8 as a promising candidate for further investigation in the treatment of HER2-positive tumors.    Click to Show/Hide
Description	To evaluated the in vitro anti-tumor activity of the conjugates, HER2-positive SK-BR-3, NCI-N87, HER2-negative MDA-MB-231 and normal MCF-10A cells were selected. As shown in Table 2, nearly all conjugates exhibited obvious anti-proliferation activity against SK-BR-3 cells, while most of the conjugates, except for Z6 and Z9, demonstrated different degrees of inhibitory activity on NCI-N87 cells. Taken together, Z8 and Z11 exhibited superior anti-tumor activity. The antiproliferative activity of Z8 against NCI-N87 cells (IC50 = 1.91 ± 0.71 uM) was comparable to that of CPT (IC50 = 2.10 ± 1.34 uM), while the antiproliferative activity of Z11 against SK-BR-3 cells (IC50 = 0.41 ± 0.13 uM) was comparable to that of CPT (IC50 = 0.26 ± 0.06 uM). Additionally, the IC50 values of these two conjugates against MDA-MB-231 and MCF-10A cells were significantly increased, showing their ability to selectively inhibit tumor cells. Based on the above results, Z8 and Z11, which exhibited better activity and apparent selectivity, were selected for subsequent studies.    Click to Show/Hide
In Vitro Model	Gastric tubular adenocarcinoma	NCI-N87 cell		CVCL_1603
Experiment 3 Reporting the Activity Data of This PDC					[6]
Indication	Breast cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	19.10 ± 2.94 µM
Administration Time	48 h
Evaluation Method	CCK-8 assay
MOA of PDC	In the previous study, we focused on the discovery of novel cell targeting peptides, leading to the identification of the preferred HER2-targeting peptide P1 (NPNWGRSWYNQRFK) derived from pertuzumab, with the Kd of 5.70 10-6 M. Here, we designed and synthesized a series of PDCs choosing Camptothecin (CPT) as the payload and P1 as the peptide carrier. We aimed to discover novel PDC molecules with potent inhibitory activity and selectivity targeting HER2 for effective cancer treatment. Conjugate Z8, which balanced the activity and the biosafety, showed good antiproliferative activity (IC50 = 1.91 ± 0.71 M), comparable to that of CPT (IC50 = 2.10 ± 1.34 M), against NCI-N87 cells. Moreover, subsequent verification tests indicated that Z8 operated through a mechanism similar to CPT, but the results from in vivo anti-tumor activity assay suggested that Z8 exhibited greater safety compared to CPT to some extent. These findings highlight Z8 as a promising candidate for further investigation in the treatment of HER2-positive tumors.    Click to Show/Hide
Description	To evaluated the in vitro anti-tumor activity of the conjugates, HER2-positive SK-BR-3, NCI-N87, HER2-negative MDA-MB-231 and normal MCF-10A cells were selected. As shown in Table 2, nearly all conjugates exhibited obvious anti-proliferation activity against SK-BR-3 cells, while most of the conjugates, except for Z6 and Z9, demonstrated different degrees of inhibitory activity on NCI-N87 cells. Taken together, Z8 and Z11 exhibited superior anti-tumor activity. The antiproliferative activity of Z8 against NCI-N87 cells (IC50 = 1.91 ± 0.71 uM) was comparable to that of CPT (IC50 = 2.10 ± 1.34 uM), while the antiproliferative activity of Z11 against SK-BR-3 cells (IC50 = 0.41 ± 0.13 uM) was comparable to that of CPT (IC50 = 0.26 ± 0.06 uM). Additionally, the IC50 values of these two conjugates against MDA-MB-231 and MCF-10A cells were significantly increased, showing their ability to selectively inhibit tumor cells. Based on the above results, Z8 and Z11, which exhibited better activity and apparent selectivity, were selected for subsequent studies.    Click to Show/Hide
In Vitro Model	Breast adenocarcinoma	MDA-MB-231 cell		CVCL_0062
Experiment 4 Reporting the Activity Data of This PDC					[6]
Indication	Breast cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	24.45 ± 2.04 µM
Administration Time	48 h
Evaluation Method	CCK-8 assay
MOA of PDC	In the previous study, we focused on the discovery of novel cell targeting peptides, leading to the identification of the preferred HER2-targeting peptide P1 (NPNWGRSWYNQRFK) derived from pertuzumab, with the Kd of 5.70 10-6 M. Here, we designed and synthesized a series of PDCs choosing Camptothecin (CPT) as the payload and P1 as the peptide carrier. We aimed to discover novel PDC molecules with potent inhibitory activity and selectivity targeting HER2 for effective cancer treatment. Conjugate Z8, which balanced the activity and the biosafety, showed good antiproliferative activity (IC50 = 1.91 ± 0.71 M), comparable to that of CPT (IC50 = 2.10 ± 1.34 M), against NCI-N87 cells. Moreover, subsequent verification tests indicated that Z8 operated through a mechanism similar to CPT, but the results from in vivo anti-tumor activity assay suggested that Z8 exhibited greater safety compared to CPT to some extent. These findings highlight Z8 as a promising candidate for further investigation in the treatment of HER2-positive tumors.    Click to Show/Hide
Description	To evaluated the in vitro anti-tumor activity of the conjugates, HER2-positive SK-BR-3, NCI-N87, HER2-negative MDA-MB-231 and normal MCF-10A cells were selected. As shown in Table 2, nearly all conjugates exhibited obvious anti-proliferation activity against SK-BR-3 cells, while most of the conjugates, except for Z6 and Z9, demonstrated different degrees of inhibitory activity on NCI-N87 cells. Taken together, Z8 and Z11 exhibited superior anti-tumor activity. The antiproliferative activity of Z8 against NCI-N87 cells (IC50 = 1.91 ± 0.71 uM) was comparable to that of CPT (IC50 = 2.10 ± 1.34 uM), while the antiproliferative activity of Z11 against SK-BR-3 cells (IC50 = 0.41 ± 0.13 uM) was comparable to that of CPT (IC50 = 0.26 ± 0.06 uM). Additionally, the IC50 values of these two conjugates against MDA-MB-231 and MCF-10A cells were significantly increased, showing their ability to selectively inhibit tumor cells. Based on the above results, Z8 and Z11, which exhibited better activity and apparent selectivity, were selected for subsequent studies.    Click to Show/Hide
In Vitro Model	Normal	MCF-10A cell		CVCL_0598

Experiment 1 Reporting the Activity Data of This PDC					[13]
Indication	Neuroblastoma
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	2.47 ± 0.91 µM
Administration Time	72 h
Evaluation Method	CellTiter-Glo luminescent cell viability assay
Description	The cytotoxic effect of the peptide-drug conjugate on Kelly-WT cells was comparable to that of 9, but not as strong as the one of the native drug. Contrary to the other utilized substances, the antiproliferative action of the bioconjugate in wild-type and drug-resistant cells was nearly the same. In comparison with doxorubicin the cytotoxicity of 10 against Kelly-ADR cells was increased by a factor of 3 according to the obtained IC50 values. Even the unmodified dimer had roughly the same cytotoxic effect on Kelly-ADR cells as doxorubicin.    Click to Show/Hide
In Vitro Model	Neuroblastoma	Kelly-WT cell		CVCL_2092
Experiment 2 Reporting the Activity Data of This PDC					[13]
Indication	Neuroblastoma
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	2.76 ± 0.33 µM
Administration Time	72 h
Evaluation Method	CellTiter-Glo luminescent cell viability assay
Description	The cytotoxic effect of the peptide-drug conjugate on Kelly-WT cells was comparable to that of 9, but not as strong as the one of the native drug. Contrary to the other utilized substances, the antiproliferative action of the bioconjugate in wild-type and drug-resistant cells was nearly the same. In comparison with doxorubicin the cytotoxicity of 10 against Kelly-ADR cells was increased by a factor of 3 according to the obtained IC50 values. Even the unmodified dimer had roughly the same cytotoxic effect on Kelly-ADR cells as doxorubicin.    Click to Show/Hide
In Vitro Model	Neuroblastoma	Kelly-ADR cell		CVCL_2092
Experiment 3 Reporting the Activity Data of This PDC					[13]
Indication	Neuroblastoma
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	3.66 ± 0.77 µM
Administration Time	72 h
Evaluation Method	CellTiter-Glo luminescent cell viability assay
Description	The obtained IC50 values for 9 (6.73 ± 2.44 uM) and 10 (3.66 ± 0.77 uM) were higher than for doxorubicin (0.90 ± 0.12 uM).
In Vitro Model	Invasive breast carcinoma	MCF-7 cell		CVCL_0031

Experiment 1 Reporting the Activity Data of This PDC					[14]
Indication	Pancreatic cancer
Efficacy Data	Half maximal inhibitory concentration (IC50)	5.0 ±0.6 μM
Administration Time	72 h
Evaluation Method	MTS assay
MOA of PDC	Towards these ends, we report on the design of a modular and user-friendly approach to enable bio-orthogonal conjugation of synthetic amanitins to three different linkers that are elaborated to three potent peptide-amanitin conjugates that differ by their mode of toxin release, as defined by the linker. Octreotate (TATE-N3, 2) was chosen as it is a circulation-stable octapeptide somatostatin analog that agonizes somatostatin receptors (Kd &tide; 0.4 nM for sstr2). Targeting sstr2 has been successfully exploited for drug delivery, cancer imaging, and radiotherapy owing to the high levels of sstr2 expression on neuroendocrine tumors (e.g. carcinoids, pancreatic islet cell tumors, thyroid carcinomas, and small lung cancer to name a few). While this is the first report of an octreotate-amanitin conjugate, herein, octreotate serves as an exemplar to highlight the potential for developing peptide-amanitin conjugates in targeted applications.    Click to Show/Hide
Description	With bioconjugates 5-7 in hand, we evaluated their cytotoxicity by assaying cell viability on an sstr2-positive rat pancreatic cancer Ar42J cells, using free amatoxins as controls. The MTS cell viability assay showed that all three PDCs were effective at reducing the viability of Ar42J cells in the low nM range, providing calculated EC50 values of 4.2, and 2.3 nM for conjugates 6 and 7, respectively; these displayed up to 1000-fold enhancement in apparent cytotoxicity compared to free amatoxins 3 and 4, both of which gave calculated EC50 values of &tide;2 μM. This points to successful targeting and intracellular accumulation of toxin inside cells through an sstr2-mediated uptake inside the cells. Further evidence supporting target-mediated uptake is the micromolar toxicity of bioconjugates to the sstr2-negative CHO cell line that is otherwise sensitive to non-targeted amanitin. In comparison to -amanitin and synthetic amatoxins 3 and 4, bioconjugates 5-7 are 7- to 14-fold less toxic to CHO cells (control cell line), likely owing to impaired uptake of octreotate-amanitin conjugates that must enter via diffusion mediated processes or other non-specific import mechanisms that are currently unknown. In addition, a blocking study was run in the presence of free TATE-N32, which led to the expected reduction in apparent toxicity.    Click to Show/Hide
In Vitro Model	Digestive system neoplasms	AR42J (SSTR2+) cell		CVCL_0143
Experiment 2 Reporting the Activity Data of This PDC					[14]
Indication	Pancreatic cancer
Efficacy Data	Half maximal effective concentration (EC50)	4.2 ±0.6 nM
Administration Time	72 h
Evaluation Method	MTS assay
MOA of PDC	Towards these ends, we report on the design of a modular and user-friendly approach to enable bio-orthogonal conjugation of synthetic amanitins to three different linkers that are elaborated to three potent peptide-amanitin conjugates that differ by their mode of toxin release, as defined by the linker. Octreotate (TATE-N3, 2) was chosen as it is a circulation-stable octapeptide somatostatin analog that agonizes somatostatin receptors (Kd &tide; 0.4 nM for sstr2). Targeting sstr2 has been successfully exploited for drug delivery, cancer imaging, and radiotherapy owing to the high levels of sstr2 expression on neuroendocrine tumors (e.g. carcinoids, pancreatic islet cell tumors, thyroid carcinomas, and small lung cancer to name a few). While this is the first report of an octreotate-amanitin conjugate, herein, octreotate serves as an exemplar to highlight the potential for developing peptide-amanitin conjugates in targeted applications.    Click to Show/Hide
Description	With bioconjugates 5-7 in hand, we evaluated their cytotoxicity by assaying cell viability on an sstr2-positive rat pancreatic cancer Ar42J cells, using free amatoxins as controls. The MTS cell viability assay showed that all three PDCs were effective at reducing the viability of Ar42J cells in the low nM range, providing calculated EC50 values of 4.2, and 2.3 nM for conjugates 6 and 7, respectively; these displayed up to 1000-fold enhancement in apparent cytotoxicity compared to free amatoxins 3 and 4, both of which gave calculated EC50 values of &tide;2 μM. This points to successful targeting and intracellular accumulation of toxin inside cells through an sstr2-mediated uptake inside the cells. Further evidence supporting target-mediated uptake is the micromolar toxicity of bioconjugates to the sstr2-negative CHO cell line that is otherwise sensitive to non-targeted amanitin. In comparison to -amanitin and synthetic amatoxins 3 and 4, bioconjugates 5-7 are 7- to 14-fold less toxic to CHO cells (control cell line), likely owing to impaired uptake of octreotate-amanitin conjugates that must enter via diffusion mediated processes or other non-specific import mechanisms that are currently unknown. In addition, a blocking study was run in the presence of free TATE-N32, which led to the expected reduction in apparent toxicity.    Click to Show/Hide
In Vitro Model	Digestive system neoplasms	AR42J (SSTR2+) cell		CVCL_0143

Experiment 1 Reporting the Activity Data of This PDC					[6]
Indication	Breast cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	5.96 ± 2.41 µM
Administration Time	48 h
Evaluation Method	CCK-8 assay
MOA of PDC	In the previous study, we focused on the discovery of novel cell targeting peptides, leading to the identification of the preferred HER2-targeting peptide P1 (NPNWGRSWYNQRFK) derived from pertuzumab, with the Kd of 5.70 10-6 M. Here, we designed and synthesized a series of PDCs choosing Camptothecin (CPT) as the payload and P1 as the peptide carrier. We aimed to discover novel PDC molecules with potent inhibitory activity and selectivity targeting HER2 for effective cancer treatment. Conjugate Z8, which balanced the activity and the biosafety, showed good antiproliferative activity (IC50 = 1.91 ± 0.71 M), comparable to that of CPT (IC50 = 2.10 ± 1.34 M), against NCI-N87 cells. Moreover, subsequent verification tests indicated that Z8 operated through a mechanism similar to CPT, but the results from in vivo anti-tumor activity assay suggested that Z8 exhibited greater safety compared to CPT to some extent. These findings highlight Z8 as a promising candidate for further investigation in the treatment of HER2-positive tumors.    Click to Show/Hide
Description	To evaluated the in vitro anti-tumor activity of the conjugates, HER2-positive SK-BR-3, NCI-N87, HER2-negative MDA-MB-231 and normal MCF-10A cells were selected. As shown in Table 2, nearly all conjugates exhibited obvious anti-proliferation activity against SK-BR-3 cells, while most of the conjugates, except for Z6 and Z9, demonstrated different degrees of inhibitory activity on NCI-N87 cells. Taken together, Z8 and Z11 exhibited superior anti-tumor activity. The antiproliferative activity of Z8 against NCI-N87 cells (IC50 = 1.91 ± 0.71 uM) was comparable to that of CPT (IC50 = 2.10 ± 1.34 uM), while the antiproliferative activity of Z11 against SK-BR-3 cells (IC50 = 0.41 ± 0.13 uM) was comparable to that of CPT (IC50 = 0.26 ± 0.06 uM). Additionally, the IC50 values of these two conjugates against MDA-MB-231 and MCF-10A cells were significantly increased, showing their ability to selectively inhibit tumor cells. Based on the above results, Z8 and Z11, which exhibited better activity and apparent selectivity, were selected for subsequent studies.    Click to Show/Hide
In Vitro Model	Breast adenocarcinoma	SK-BR-3 cell		CVCL_0033
Experiment 2 Reporting the Activity Data of This PDC					[6]
Indication	Gastric cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	10.23 ± 1.65 µM
Administration Time	48 h
Evaluation Method	CCK-8 assay
MOA of PDC	In the previous study, we focused on the discovery of novel cell targeting peptides, leading to the identification of the preferred HER2-targeting peptide P1 (NPNWGRSWYNQRFK) derived from pertuzumab, with the Kd of 5.70 10-6 M. Here, we designed and synthesized a series of PDCs choosing Camptothecin (CPT) as the payload and P1 as the peptide carrier. We aimed to discover novel PDC molecules with potent inhibitory activity and selectivity targeting HER2 for effective cancer treatment. Conjugate Z8, which balanced the activity and the biosafety, showed good antiproliferative activity (IC50 = 1.91 ± 0.71 M), comparable to that of CPT (IC50 = 2.10 ± 1.34 M), against NCI-N87 cells. Moreover, subsequent verification tests indicated that Z8 operated through a mechanism similar to CPT, but the results from in vivo anti-tumor activity assay suggested that Z8 exhibited greater safety compared to CPT to some extent. These findings highlight Z8 as a promising candidate for further investigation in the treatment of HER2-positive tumors.    Click to Show/Hide
Description	To evaluated the in vitro anti-tumor activity of the conjugates, HER2-positive SK-BR-3, NCI-N87, HER2-negative MDA-MB-231 and normal MCF-10A cells were selected. As shown in Table 2, nearly all conjugates exhibited obvious anti-proliferation activity against SK-BR-3 cells, while most of the conjugates, except for Z6 and Z9, demonstrated different degrees of inhibitory activity on NCI-N87 cells. Taken together, Z8 and Z11 exhibited superior anti-tumor activity. The antiproliferative activity of Z8 against NCI-N87 cells (IC50 = 1.91 ± 0.71 uM) was comparable to that of CPT (IC50 = 2.10 ± 1.34 uM), while the antiproliferative activity of Z11 against SK-BR-3 cells (IC50 = 0.41 ± 0.13 uM) was comparable to that of CPT (IC50 = 0.26 ± 0.06 uM). Additionally, the IC50 values of these two conjugates against MDA-MB-231 and MCF-10A cells were significantly increased, showing their ability to selectively inhibit tumor cells. Based on the above results, Z8 and Z11, which exhibited better activity and apparent selectivity, were selected for subsequent studies.    Click to Show/Hide
In Vitro Model	Gastric tubular adenocarcinoma	NCI-N87 cell		CVCL_1603
Experiment 3 Reporting the Activity Data of This PDC					[6]
Indication	Breast cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	15.44 ± 2.41 µM
Administration Time	48 h
Evaluation Method	CCK-8 assay
MOA of PDC	In the previous study, we focused on the discovery of novel cell targeting peptides, leading to the identification of the preferred HER2-targeting peptide P1 (NPNWGRSWYNQRFK) derived from pertuzumab, with the Kd of 5.70 10-6 M. Here, we designed and synthesized a series of PDCs choosing Camptothecin (CPT) as the payload and P1 as the peptide carrier. We aimed to discover novel PDC molecules with potent inhibitory activity and selectivity targeting HER2 for effective cancer treatment. Conjugate Z8, which balanced the activity and the biosafety, showed good antiproliferative activity (IC50 = 1.91 ± 0.71 M), comparable to that of CPT (IC50 = 2.10 ± 1.34 M), against NCI-N87 cells. Moreover, subsequent verification tests indicated that Z8 operated through a mechanism similar to CPT, but the results from in vivo anti-tumor activity assay suggested that Z8 exhibited greater safety compared to CPT to some extent. These findings highlight Z8 as a promising candidate for further investigation in the treatment of HER2-positive tumors.    Click to Show/Hide
Description	To evaluated the in vitro anti-tumor activity of the conjugates, HER2-positive SK-BR-3, NCI-N87, HER2-negative MDA-MB-231 and normal MCF-10A cells were selected. As shown in Table 2, nearly all conjugates exhibited obvious anti-proliferation activity against SK-BR-3 cells, while most of the conjugates, except for Z6 and Z9, demonstrated different degrees of inhibitory activity on NCI-N87 cells. Taken together, Z8 and Z11 exhibited superior anti-tumor activity. The antiproliferative activity of Z8 against NCI-N87 cells (IC50 = 1.91 ± 0.71 uM) was comparable to that of CPT (IC50 = 2.10 ± 1.34 uM), while the antiproliferative activity of Z11 against SK-BR-3 cells (IC50 = 0.41 ± 0.13 uM) was comparable to that of CPT (IC50 = 0.26 ± 0.06 uM). Additionally, the IC50 values of these two conjugates against MDA-MB-231 and MCF-10A cells were significantly increased, showing their ability to selectively inhibit tumor cells. Based on the above results, Z8 and Z11, which exhibited better activity and apparent selectivity, were selected for subsequent studies.    Click to Show/Hide
In Vitro Model	Breast adenocarcinoma	MDA-MB-231 cell		CVCL_0062
Experiment 4 Reporting the Activity Data of This PDC					[6]
Indication	Breast cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	33.12 ± 3.84 µM
Administration Time	48 h
Evaluation Method	CCK-8 assay
MOA of PDC	In the previous study, we focused on the discovery of novel cell targeting peptides, leading to the identification of the preferred HER2-targeting peptide P1 (NPNWGRSWYNQRFK) derived from pertuzumab, with the Kd of 5.70 10-6 M. Here, we designed and synthesized a series of PDCs choosing Camptothecin (CPT) as the payload and P1 as the peptide carrier. We aimed to discover novel PDC molecules with potent inhibitory activity and selectivity targeting HER2 for effective cancer treatment. Conjugate Z8, which balanced the activity and the biosafety, showed good antiproliferative activity (IC50 = 1.91 ± 0.71 M), comparable to that of CPT (IC50 = 2.10 ± 1.34 M), against NCI-N87 cells. Moreover, subsequent verification tests indicated that Z8 operated through a mechanism similar to CPT, but the results from in vivo anti-tumor activity assay suggested that Z8 exhibited greater safety compared to CPT to some extent. These findings highlight Z8 as a promising candidate for further investigation in the treatment of HER2-positive tumors.    Click to Show/Hide
Description	To evaluated the in vitro anti-tumor activity of the conjugates, HER2-positive SK-BR-3, NCI-N87, HER2-negative MDA-MB-231 and normal MCF-10A cells were selected. As shown in Table 2, nearly all conjugates exhibited obvious anti-proliferation activity against SK-BR-3 cells, while most of the conjugates, except for Z6 and Z9, demonstrated different degrees of inhibitory activity on NCI-N87 cells. Taken together, Z8 and Z11 exhibited superior anti-tumor activity. The antiproliferative activity of Z8 against NCI-N87 cells (IC50 = 1.91 ± 0.71 uM) was comparable to that of CPT (IC50 = 2.10 ± 1.34 uM), while the antiproliferative activity of Z11 against SK-BR-3 cells (IC50 = 0.41 ± 0.13 uM) was comparable to that of CPT (IC50 = 0.26 ± 0.06 uM). Additionally, the IC50 values of these two conjugates against MDA-MB-231 and MCF-10A cells were significantly increased, showing their ability to selectively inhibit tumor cells. Based on the above results, Z8 and Z11, which exhibited better activity and apparent selectivity, were selected for subsequent studies.    Click to Show/Hide
In Vitro Model	Normal	MCF-10A cell		CVCL_0598

Experiment 1 Reporting the Activity Data of This PDC					[15]
Indication	Tumor
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	6.21 ± 1.12 µM
Evaluation Method	CCK-8 assay
Description	The BP9a-SS-DOX conjugate exhibited impressed antiproliferative activity against HepG2 cells with an IC50 value of 6.21 ± 1.12 uM which was lower than that of free DOX.
In Vitro Model	Hepatoblastoma	Hep-G2 cell		CVCL_0027

Experiment 1 Reporting the Activity Data of This PDC					[16]
Indication	Breast cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	11.4 μM
Description	Doxorubicin binds with high affinity to DNA by intercalation, which leads to DNA damage and subsequent growth inhibition44,45. We performed MTT assays in order to determine the viability of the tumor cells and additionally to assess the proliferative potential of the cells after drug treatment. MCF-7 and HT-29 cells were incubated with doxorubicin and the peptide conjugates at various concentrations in the range of 0.1 and 50 μmfor 72 h (in a serum-containing medium), and the results were expressed as IC50-values (Table 1).    Click to Show/Hide
In Vitro Model	Invasive breast carcinoma	MCF-7 cell		CVCL_0031
Experiment 2 Reporting the Activity Data of This PDC					[16]
Indication	Breast cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	27 μM
Description	Doxorubicin binds with high affinity to DNA by intercalation, which leads to DNA damage and subsequent growth inhibition44,45. We performed MTT assays in order to determine the viability of the tumor cells and additionally to assess the proliferative potential of the cells after drug treatment. MCF-7 and HT-29 cells were incubated with doxorubicin and the peptide conjugates at various concentrations in the range of 0.1 and 50 μmfor 72 h (in a serum-containing medium), and the results were expressed as IC50-values (Table 1).    Click to Show/Hide
In Vitro Model	Invasive breast carcinoma	MCF-7 cell		CVCL_0031

Experiment 1 Reporting the Activity Data of This PDC					[16]
Indication	Breast cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	19 μM
Description	Doxorubicin binds with high affinity to DNA by intercalation, which leads to DNA damage and subsequent growth inhibition44,45. We performed MTT assays in order to determine the viability of the tumor cells and additionally to assess the proliferative potential of the cells after drug treatment. MCF-7 and HT-29 cells were incubated with doxorubicin and the peptide conjugates at various concentrations in the range of 0.1 and 50 μmfor 72 h (in a serum-containing medium), and the results were expressed as IC50-values (Table 1).    Click to Show/Hide
In Vitro Model	Colon cancer	HT29 cell		CVCL_A8EZ
Experiment 2 Reporting the Activity Data of This PDC					[16]
Indication	Breast cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	24.7 μM
Description	Doxorubicin binds with high affinity to DNA by intercalation, which leads to DNA damage and subsequent growth inhibition44,45. We performed MTT assays in order to determine the viability of the tumor cells and additionally to assess the proliferative potential of the cells after drug treatment. MCF-7 and HT-29 cells were incubated with doxorubicin and the peptide conjugates at various concentrations in the range of 0.1 and 50 μmfor 72 h (in a serum-containing medium), and the results were expressed as IC50-values (Table 1).    Click to Show/Hide
In Vitro Model	Colon cancer	HT29 cell		CVCL_A8EZ

Experiment 1 Reporting the Activity Data of This PDC					[17]
Indication	Breast cancer
Efficacy Data	Half Maximal Inhibitory Concentration (IC50)	> 400 µM
In Vitro Model	Invasive breast carcinoma	MCF-7 cell		CVCL_0031

Experiment 1 Reporting the Activity Data of This PDC					[18]
Indication	Tumor
Efficacy Data	Cell viability	18%
Administration Time	72 h
Administration Dosage	10 µM
Evaluation Method	MTS assay
Description	HeLa cells treated with pHLIP-S-S-DOX at low pH showed significant toxicity (˜50% viability at the lowest concentration tested (0.16 μM pHLIP)) which, in this construct, corresponds to the DOX concentration. Cellular viability tracked inversely with increasing pHLIP conjugate concentration, peaking at ˜18% viability at 16 μM.
In Vitro Model	Endocervical adenocarcinoma	HeLa cell		CVCL_0030
Experiment 2 Reporting the Activity Data of This PDC					[18]
Indication	Tumor
Efficacy Data	Cell viability	20%
Administration Time	72 h
Administration Dosage	1.3 µM
Evaluation Method	MTS assay
Description	HeLa cells treated with pHLIP-S-S-DOX at low pH showed significant toxicity (˜50% viability at the lowest concentration tested (0.16 μM pHLIP)) which, in this construct, corresponds to the DOX concentration. Cellular viability tracked inversely with increasing pHLIP conjugate concentration, peaking at ˜18% viability at 13 μM.
In Vitro Model	Endocervical adenocarcinoma	HeLa cell		CVCL_0030
Experiment 3 Reporting the Activity Data of This PDC					[18]
Indication	Tumor
Efficacy Data	Cell viability	22%
Administration Time	72 h
Administration Dosage	2.5 µM
Evaluation Method	MTS assay
Description	HeLa cells treated with pHLIP-S-S-DOX at low pH showed significant toxicity (˜50% viability at the lowest concentration tested (0.16 μM pHLIP)) which, in this construct, corresponds to the DOX concentration. Cellular viability tracked inversely with increasing pHLIP conjugate concentration, peaking at ˜18% viability at 14 μM.
In Vitro Model	Endocervical adenocarcinoma	HeLa cell		CVCL_0030
Experiment 4 Reporting the Activity Data of This PDC					[18]
Indication	Tumor
Efficacy Data	Cell viability	22%
Administration Time	72 h
Administration Dosage	5.0 µM
Evaluation Method	MTS assay
Description	HeLa cells treated with pHLIP-S-S-DOX at low pH showed significant toxicity (˜50% viability at the lowest concentration tested (0.16 μM pHLIP)) which, in this construct, corresponds to the DOX concentration. Cellular viability tracked inversely with increasing pHLIP conjugate concentration, peaking at ˜18% viability at 15 μM.
In Vitro Model	Endocervical adenocarcinoma	HeLa cell		CVCL_0030
Experiment 5 Reporting the Activity Data of This PDC					[18]
Indication	Tumor
Efficacy Data	Cell viability	30%
Administration Time	72 h
Administration Dosage	0.31 µM
Evaluation Method	MTS assay
Description	The cytotoxicity data for the pHLIP-PAMAM-DOX conjugate mirrored quite closely that of the pHLIP-S-S-DOX conjugate, particularly at the higher concentrations (>1.25 μM). However, at lower pHLIP concentrations (0.16 μM-0.63 μM), the PAMAM conjugate exhibited higher cytotoxicity than the single DOX conjugate (˜up to 17% higher toxicity).    Click to Show/Hide
In Vitro Model	Endocervical adenocarcinoma	HeLa cell		CVCL_0030
Experiment 6 Reporting the Activity Data of This PDC					[18]
Indication	Tumor
Efficacy Data	Cell viability	30%
Administration Time	72 h
Administration Dosage	0.63 µM
Evaluation Method	MTS assay
Description	The cytotoxicity data for the pHLIP-PAMAM-DOX conjugate mirrored quite closely that of the pHLIP-S-S-DOX conjugate, particularly at the higher concentrations (>1.25 μM). However, at lower pHLIP concentrations (0.16 μM-0.63 μM), the PAMAM conjugate exhibited higher cytotoxicity than the single DOX conjugate (˜up to 17% higher toxicity).    Click to Show/Hide
In Vitro Model	Endocervical adenocarcinoma	HeLa cell		CVCL_0030
Experiment 7 Reporting the Activity Data of This PDC					[18]
Indication	Tumor
Efficacy Data	Cell viability	30%
Administration Time	72 h
Administration Dosage	1.3 µM
Evaluation Method	MTS assay
Description	The cytotoxicity data for the pHLIP-PAMAM-DOX conjugate mirrored quite closely that of the pHLIP-S-S-DOX conjugate, particularly at the higher concentrations (>1.25 μM). However, at lower pHLIP concentrations (0.16 μM-0.63 μM), the PAMAM conjugate exhibited higher cytotoxicity than the single DOX conjugate (˜up to 17% higher toxicity).    Click to Show/Hide
In Vitro Model	Endocervical adenocarcinoma	HeLa cell		CVCL_0030
Experiment 8 Reporting the Activity Data of This PDC					[18]
Indication	Tumor
Efficacy Data	Cell viability	38%
Administration Time	72 h
Administration Dosage	0.16 µM
Evaluation Method	MTS assay
Description	The cytotoxicity data for the pHLIP-PAMAM-DOX conjugate mirrored quite closely that of the pHLIP-S-S-DOX conjugate, particularly at the higher concentrations (>1.25 μM). However, at lower pHLIP concentrations (0.16 μM-0.63 μM), the PAMAM conjugate exhibited higher cytotoxicity than the single DOX conjugate (˜up to 17% higher toxicity).    Click to Show/Hide
In Vitro Model	Endocervical adenocarcinoma	HeLa cell		CVCL_0030
Experiment 9 Reporting the Activity Data of This PDC					[18]
Indication	Tumor
Efficacy Data	Cell viability	42%
Administration Time	72 h
Administration Dosage	0.63 µM
Evaluation Method	MTS assay
Description	HeLa cells treated with pHLIP-S-S-DOX at low pH showed significant toxicity (˜50% viability at the lowest concentration tested (0.16 μM pHLIP)) which, in this construct, corresponds to the DOX concentration. Cellular viability tracked inversely with increasing pHLIP conjugate concentration, peaking at ˜18% viability at 12 μM.
In Vitro Model	Endocervical adenocarcinoma	HeLa cell		CVCL_0030
Experiment 10 Reporting the Activity Data of This PDC					[18]
Indication	Tumor
Efficacy Data	Cell viability	45%
Administration Time	72 h
Administration Dosage	0.31 µM
Evaluation Method	MTS assay
Description	HeLa cells treated with pHLIP-S-S-DOX at low pH showed significant toxicity (˜50% viability at the lowest concentration tested (0.16 μM pHLIP)) which, in this construct, corresponds to the DOX concentration. Cellular viability tracked inversely with increasing pHLIP conjugate concentration, peaking at ˜18% viability at 11 μM.
In Vitro Model	Endocervical adenocarcinoma	HeLa cell		CVCL_0030
Experiment 11 Reporting the Activity Data of This PDC					[18]
Indication	Tumor
Efficacy Data	Cell viability	50%
Administration Time	72 h
Administration Dosage	0.16 µM
Evaluation Method	MTS assay
Description	HeLa cells treated with pHLIP-S-S-DOX at low pH showed significant toxicity (˜50% viability at the lowest concentration tested (0.16 μM pHLIP)) which, in this construct, corresponds to the DOX concentration. Cellular viability tracked inversely with increasing pHLIP conjugate concentration, peaking at ˜18% viability at 10 μM.
In Vitro Model	Endocervical adenocarcinoma	HeLa cell		CVCL_0030
Experiment 12 Reporting the Activity Data of This PDC					[18]
Indication	Tumor
Efficacy Data	Cell viability	90%
Administration Time	72 h
Administration Dosage	0.63 µM
Evaluation Method	MTS assay
Description	HeLa cells treated with pHLIP-S-S-DOX at low pH showed significant toxicity (˜50% viability at the lowest concentration tested (0.16 μM pHLIP)) which, in this construct, corresponds to the DOX concentration. Cellular viability tracked inversely with increasing pHLIP conjugate concentration, peaking at ˜18% viability at 19 μM.
In Vitro Model	Endocervical adenocarcinoma	HeLa cell		CVCL_0030
Experiment 13 Reporting the Activity Data of This PDC					[18]
Indication	Tumor
Efficacy Data	Cell viability	95%
Administration Time	72 h
Administration Dosage	0.16 µM
Evaluation Method	MTS assay
Description	HeLa cells treated with pHLIP-S-S-DOX at low pH showed significant toxicity (˜50% viability at the lowest concentration tested (0.16 μM pHLIP)) which, in this construct, corresponds to the DOX concentration. Cellular viability tracked inversely with increasing pHLIP conjugate concentration, peaking at ˜18% viability at 17 μM.
In Vitro Model	Endocervical adenocarcinoma	HeLa cell		CVCL_0030
Experiment 14 Reporting the Activity Data of This PDC					[18]
Indication	Tumor
Efficacy Data	Cell viability	98%
Administration Time	72 h
Administration Dosage	0.31 µM
Evaluation Method	MTS assay
Description	HeLa cells treated with pHLIP-S-S-DOX at low pH showed significant toxicity (˜50% viability at the lowest concentration tested (0.16 μM pHLIP)) which, in this construct, corresponds to the DOX concentration. Cellular viability tracked inversely with increasing pHLIP conjugate concentration, peaking at ˜18% viability at 18 μM.
In Vitro Model	Endocervical adenocarcinoma	HeLa cell		CVCL_0030
Experiment 15 Reporting the Activity Data of This PDC					[18]
Indication	Tumor
Efficacy Data	Cell viability	99%
Administration Time	72 h
Administration Dosage	10 µM
Evaluation Method	MTS assay
Description	HeLa cells treated with pHLIP-S-S-DOX at low pH showed significant toxicity (˜50% viability at the lowest concentration tested (0.16 μM pHLIP)) which, in this construct, corresponds to the DOX concentration. Cellular viability tracked inversely with increasing pHLIP conjugate concentration, peaking at ˜18% viability at 23 μM.
In Vitro Model	Endocervical adenocarcinoma	HeLa cell		CVCL_0030
Experiment 16 Reporting the Activity Data of This PDC					[18]
Indication	Tumor
Efficacy Data	Cell viability	100%
Administration Time	72 h
Administration Dosage	1.3 µM
Evaluation Method	MTS assay
Description	HeLa cells treated with pHLIP-S-S-DOX at low pH showed significant toxicity (˜50% viability at the lowest concentration tested (0.16 μM pHLIP)) which, in this construct, corresponds to the DOX concentration. Cellular viability tracked inversely with increasing pHLIP conjugate concentration, peaking at ˜18% viability at 20 μM.
In Vitro Model	Endocervical adenocarcinoma	HeLa cell		CVCL_0030
Experiment 17 Reporting the Activity Data of This PDC					[18]
Indication	Tumor
Efficacy Data	Cell viability	100%
Administration Time	72 h
Administration Dosage	2.5 µM
Evaluation Method	MTS assay
Description	HeLa cells treated with pHLIP-S-S-DOX at low pH showed significant toxicity (˜50% viability at the lowest concentration tested (0.16 μM pHLIP)) which, in this construct, corresponds to the DOX concentration. Cellular viability tracked inversely with increasing pHLIP conjugate concentration, peaking at ˜18% viability at 21 μM.
In Vitro Model	Endocervical adenocarcinoma	HeLa cell		CVCL_0030
Experiment 18 Reporting the Activity Data of This PDC					[18]
Indication	Tumor
Efficacy Data	Cell viability	100%
Administration Time	72 h
Administration Dosage	5.0 µM
Evaluation Method	MTS assay
Description	HeLa cells treated with pHLIP-S-S-DOX at low pH showed significant toxicity (˜50% viability at the lowest concentration tested (0.16 μM pHLIP)) which, in this construct, corresponds to the DOX concentration. Cellular viability tracked inversely with increasing pHLIP conjugate concentration, peaking at ˜18% viability at 22 μM.
In Vitro Model	Endocervical adenocarcinoma	HeLa cell		CVCL_0030

Ref 1	BT1718, a novel bicyclic peptide-maytansinoid conjugate targeting MT1-MMP for the treatment of solid tumours: Design of bicyclic peptide and linker selection.
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