Drug Information

General Information of This Drug
Drug ID DRG00013
Drug Name Gemcitabine
Synonyms
gemcitabine; 95058-81-4; 2'-Deoxy-2',2'-difluorocytidine; 2',2'-Difluorodeoxycytidine; dFdC; Cytidine, 2'-deoxy-2',2'-difluoro-; Gemcitabine free base; 2',2'-difluoro-2'-deoxycytidine; 103882-84-4; 4-amino-1-[(2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)oxolan-2-yl]-1,2-dihydropyrimidin-2-one; LY188011; 4-amino-1-((2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyrimidin-2(1H)-one; 95058-81-4 (free base); B76N6SBZ8R; LY 188011; 4-Amino-1-[(2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)oxolan-2-yl]pyrimidin-2-one; Gamcitabine; Gemcitabina; CHEBI:175901; DFdCyd; NSC-613327; Gemcitabinum; Folfugem; Gemcel; Zefei; GemLip; 4-amino-1-((2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)-tetrahydrofuran-2-yl)pyrimidin-2(1H)-one; Gemcitabinum [INN-Latin]; Gemcitabina [INN-Spanish]; C9H11F2N3O4; Gemzar (hydrochloride); SR-05000001491; Gemcitabine (USAN/INN); UNII-B76N6SBZ8R; 2',2'-DiF-dC; Gemcitabine [USAN:INN:BAN]; CCRIS 8984; MFCD00869720; HSDB 7567; 2vpp; gemcitabine (Gemzar); NSC 613327; 4pd5; GEMCITABINE [MI]; GEMCITABINE [INN]; 2'-Deoxy-.beta.-D-2',2'-difluorocytidine; GEMCITABINE [HSDB]; GEMCITABINE [USAN]; CHEMBL888; GEMCITABINE [VANDF]; SCHEMBL4295; GEMCITABINE [WHO-DD]; GTPL4793; DTXSID3040487; 2'deoxy-2',2'-difluorocytidine; BCPP000219; BDBM429521; GLXC-04598; HMS2089P10; HMS3715N07; 2`-Deoxy-2`,2`-difluorocytidine; med.21724, Compound Gemcitabine; DL-215; s1714; AKOS015920303; Cytidine, 2'-deoxy-2',2'-difluoro-2'-Deoxy-.beta.-D-2',2'-difluorocytidine; BCP9000721; CCG-221183; CS-0643; DB00441; GS-3582; 4-Amino-1-[3,3-difluoro-4-hydroxy-5-(hydroxymethyl) tetrahydrofuran-2-yl]-1H-pyrimidin-2-one; NCGC00168784-02; NCGC00168784-08; NCGC00168784-12; BP-58640; HY-17026; G0544; NS00000342; SW199649-2; C07650; D02368; EN300-267822; AB01274777-01; AB01274777-02; AB01274777_05; AB01274777_06; Q414143; J-001056; SR-05000001491-1; SR-05000001491-2; BRD-K15108141-001-04-1; Z1741982024; 4-AMINO-1-[(2R,4R,5R)-3,3-DIFLUORO-4-HYDROXY-5-(HYDROXYMETHYL)TETRAHYDRO-2-FURANYL]-2(1H)-PYRIMIDINONE; 4-amino-1-[(2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl]pyrimidin-2-one
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Target(s) Ribonucleoside-diphosphate reductase subunit M2 (RRM2)  Target Info 
Structure
Formula
C9H11F2N3O4
#Ro5 Violations (Lipinski): 0 Molecular Weight (mw) 263.2
Lipid-water partition coefficient (xlogp) -1.5
Hydrogen Bond Donor Count (hbonddonor) 3
Hydrogen Bond Acceptor Count (hbondacc) 6
Rotatable Bond Count (rotbonds) 2
PubChem CID
60750
Canonical smiles
C1=CN(C(=O)N=C1N)C2C(C(C(O2)CO)O)(F)F
InChI
InChI=1S/C9H11F2N3O4/c10-9(11)6(16)4(3-15)18-7(9)14-2-1-5(12)13-8(14)17/h1-2,4,6-7,15-16H,3H2,(H2,12,13,17)/t4-,6-,7-/m1/s1
InChIKey
SDUQYLNIPVEERB-QPPQHZFASA-N
IUPAC Name
4-amino-1-[(2R,4R,5R)-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)oxolan-2-yl]pyrimidin-2-one
The activity data of This Drug
Standard Type Value Disease Model Cell line Cell line ID Ref.
Half Maximal Growth Inhibition (GI50) 2 nM Lung adenocarcinoma NCI-H23 cell CVCL_1547 [1]
Half Maximal Growth Inhibition (GI50) 2.085 nM Lung large cell carcinoma NCI-H460 cell CVCL_0459 [2]
Half Maximal Growth Inhibition (GI50) 3 nM Colon adenocarcinoma HCT 15 cell CVCL_0292 [1]
Half Maximal Growth Inhibition (GI50) 4 nM Breast ductal carcinoma BT-549 cell CVCL_1092 [1]
Half Maximal Growth Inhibition (GI50) 6 nM Prostate carcinoma PC-3 cell CVCL_0035 [1]
Half Maximal Growth Inhibition (GI50) 10 nM Invasive breast carcinoma MCF-7 cell CVCL_0031 [3]
Half Maximal Inhibitory Concentration (IC50) 1.5 nM Ovarian endometrioid adenocarcinoma A2780 cell CVCL_0134 [4]
Half Maximal Inhibitory Concentration (IC50) 2.6 nM High grade ovarian serous adenocarcinoma OVCAR-8 cell CVCL_1629 [5]
Half Maximal Inhibitory Concentration (IC50) 6 nM Pancreatic ductal adenocarcinoma BxPC-3 cell CVCL_0186 [6]
Half Maximal Inhibitory Concentration (IC50) 6 nM Colon carcinoma CT26 cell CVCL_7254 [7]
Half Maximal Inhibitory Concentration (IC50) 6 nM Colon carcinoma HCT 116 cell CVCL_0291 [8]
Half Maximal Inhibitory Concentration (IC50) 7 nM Thymoma EL4 cell CVCL_0255 [7]
Half Maximal Inhibitory Concentration (IC50) 7 nM Lymphoblastic leukemia L1210 cell CVCL_0382 [7]
Half Maximal Inhibitory Concentration (IC50) 8 nM Breast ductal carcinoma BT-549 cell CVCL_1092 [7]
Half Maximal Inhibitory Concentration (IC50) 9 nM Lung small cell carcinoma DMS-53 cell CVCL_1177 [9]
Half Maximal Inhibitory Concentration (IC50) 9.2 nM Uterine sarcoma MES-SA/Dx5 cell CVCL_2598 [10]
Half Maximal Inhibitory Concentration (IC50) 9.7 nM Colon carcinoma HCT 116 cell CVCL_0291 [5]
Half Maximal Inhibitory Concentration (IC50) 9.9 nM Colon adenocarcinoma HCT 15 cell CVCL_0292 [5]
Half Maximal Inhibitory Concentration (IC50) 10.3 nM Astrocytoma SF268 cell CVCL_1689 [5]
Half Maximal Inhibitory Concentration (IC50) 11.4 nM Breast adenocarcinoma MDA-MB-231 cell CVCL_0062 [11]
Half Maximal Inhibitory Concentration (IC50) 13.6 nM Colon adenocarcinoma SW480 cell CVCL_0546 [5]
Half Maximal Inhibitory Concentration (IC50) 16 nM Pancreatic ductal adenocarcinoma MIA PaCa-2 cell CVCL_0428 [12]
Half Maximal Inhibitory Concentration (IC50) 16 nM Lung carcinoma SW1573 cell CVCL_1720 [4]
Half Maximal Inhibitory Concentration (IC50) 19 nM Pancreatic ductal adenocarcinoma Capan-1 cell CVCL_0237 [6]
Half Maximal Inhibitory Concentration (IC50) 19.8 nM Gliosarcoma SF539 cell CVCL_1691 [5]
Half Maximal Inhibitory Concentration (IC50) 20 nM Lung adenocarcinoma A-549 cell CVCL_0023 [9]
Half Maximal Inhibitory Concentration (IC50) 25 nM Breast adenocarcinoma MDA-MB-231 cell CVCL_0062 [13]
Half Maximal Inhibitory Concentration (IC50) 30 nM Colon carcinoma HCT 116 cell CVCL_0291 [14]
Half Maximal Inhibitory Concentration (IC50) 35 nM Ovarian endometrioid adenocarcinoma A2780 cell CVCL_0134 [8]
Half Maximal Inhibitory Concentration (IC50) 40 nM Prostate carcinoma PC-3 cell CVCL_0035 [15]
Half Maximal Inhibitory Concentration (IC50) 47 nM Squamous carcinoma SCC-25 cell CVCL_1682 [16]
Half Maximal Inhibitory Concentration (IC50) 50 nM Lung adenocarcinoma A-549 cell CVCL_0023 [17]
Half Maximal Inhibitory Concentration (IC50) 50 nM Chronic myeloid leukemia K562 cell CVCL_0004 [18]
Half Maximal Inhibitory Concentration (IC50) 51.4 nM Normal COLO205 cell CVCL_F402 [5]
Half Maximal Inhibitory Concentration (IC50) 70 nM Acute monocytic leukemia U-937 cell CVCL_0007 [19]
Half Maximal Inhibitory Concentration (IC50) 90.7 nM Osteosarcoma U2OS cell CVCL_0042 [5]
Half Maximal Inhibitory Concentration (IC50) 100 nM Chronic myeloid leukemia K562 cell CVCL_0004 [20]
Half Maximal Inhibitory Concentration (IC50) 120 nM Pancreatic ductal adenocarcinoma MIA PaCa-2 cell CVCL_0428 [21]
Half Maximal Inhibitory Concentration (IC50) 150 nM Pancreatic ductal adenocarcinoma PANC-1 cell CVCL_0480 [22]
Half Maximal Inhibitory Concentration (IC50) 180 nM Osteosarcoma U2OS cell CVCL_0042 [20]
Half Maximal Inhibitory Concentration (IC50) 275 nM Lung carcinoma SW1573 cell CVCL_1720 [4]
Half Maximal Inhibitory Concentration (IC50) 310 nM Ovarian endometrioid adenocarcinoma A2780 cell CVCL_0134 [23]
Half Maximal Inhibitory Concentration (IC50) 410 nM Colon carcinoma HCT 116 cell CVCL_0291 [20]
Half Maximal Inhibitory Concentration (IC50) 512 nM Prostate carcinoma LNCaP cell CVCL_0395 [7]
Half Maximal Inhibitory Concentration (IC50) 570 nM Invasive breast carcinoma MCF-7 cell CVCL_0031 [24]
Half Maximal Inhibitory Concentration (IC50) 718 nM Chronic myeloid leukemia K562 cell CVCL_0004 [7]
Half Maximal Inhibitory Concentration (IC50) 840 nM Hepatoma Bel-7402 cell CVCL_5492 [23]
Half Maximal Inhibitory Concentration (IC50) 1000 nM T acute lymphoblastic leukemia CCRF-CEM cell CVCL_0207 [4]
Half Maximal Inhibitory Concentration (IC50) 1.1 uM Neuroblastoma SK-N-AS cell CVCL_1700 [7]
Half Maximal Inhibitory Concentration (IC50) 1.4 uM Lung adenocarcinoma A-549 cell CVCL_0023 [23]
Half Maximal Inhibitory Concentration (IC50) 2.1 uM Breast adenocarcinoma MDA-MB-231 cell CVCL_0062 [25]
Half Maximal Inhibitory Concentration (IC50) 3 uM Normal COLO205 cell CVCL_F402 [8]
Half Maximal Inhibitory Concentration (IC50) 3.28 uM Invasive breast carcinoma MCF-7 cell CVCL_0031 [23]
Half Maximal Inhibitory Concentration (IC50) 4.12 uM Endocervical adenocarcinoma HeLa cell CVCL_0030 [7]
Half Maximal Inhibitory Concentration (IC50) 4.9 uM Hepatoblastoma Hep-G2 cell CVCL_0027 [26]
Half Maximal Inhibitory Concentration (IC50) 5.05 uM Lung adenocarcinoma A-549 cell CVCL_0023 [26]
Half Maximal Inhibitory Concentration (IC50) >10 uM Endocervical adenocarcinoma HeLa cell CVCL_0030 [8]
Half Maximal Inhibitory Concentration (IC50) 40.791 uM Pancreatic ductal adenocarcinoma Capan-2 cell CVCL_0026 [12]
Half Maximal Inhibitory Concentration (IC50) 50 uM T acute lymphoblastic leukemia CCRF-CEM cell CVCL_0207 [4]
Half Maximal Inhibitory Concentration (IC50) >50 uM Normal BJ cell CVCL_E483 [27]
Tumor Growth Inhibition value (TGI) 100 uM Pancreatic ductal adenocarcinoma MIA PaCa-2 cell CVCL_0428 [28]
Each Peptide-drug Conjugate Related to This Drug
Full Information of The Activity Data of The PDC(s) Related to This Drug
GOXG2 [Investigative]
 PDC Info 
Identified from the Human Clinical Data
Click To Hide/Show 1 Activity Data Related to This Level
Experiment 1 Reporting the Activity Data of This PDC [29]
Indication Prostate cancer
Efficacy Data Relative intensity 5%
Administration Time 60 min
Administration Dosage 1 μg/ml
MOA of PDC
We aimed to construct PDCs with linker controllable drug release rates simply by manipulating the linker unit. For a more rapid drug-release rate we developed GOXG1 and GOXG2. These conjugates bear a carboxylate ester linker directly attached to the primary and the secondary alcohol group of the drug respectively, followed by oxime and amide bond. The primary alcohol of gemcitabine has been used since it is involved in the phosphorylation process, through which gemcitabine exerts its cytotoxic effect. Therefore, we aimed to block the primary alcohol and examine its effect (GOXG1) and also take advantage of the secondary alcohol which could lead to a PDC with a completely different profile (GOXG2), although they share structural similarities. For a slower drug release rate, we designed and developed the PDC GN4OXG that contains an amide bond on the 4-N position of the parent drug followed by click oxime ligation and another amide bond. The stability of this molecule should be enhanced since it is devoid of rapidly hydrolyzable ester bonds. Furthermore, in this PDC since the 4-NH2 moiety of gemcitabine is capped it could further surmount the rapid gemcitabine metabolism that leads to the formation of dFdU, after the enzymatic 4-N deamination of gemcitabine by cytidine deaminase (CDA).

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Description
GOXG2 was the least stable, followed by GOXG1, while GN4OXG was the most stable.
In Vivo Model Human plasma.
Revealed Based on the Cell Line Data
Click To Hide/Show 3 Activity Data Related to This Level
Experiment 1 Reporting the Activity Data of This PDC [29]
Indication Prostate cancer
Efficacy Data Relative intensity 0%
Administration Time 10 h
Administration Dosage 1 μM
MOA of PDC
We aimed to construct PDCs with linker controllable drug release rates simply by manipulating the linker unit. For a more rapid drug-release rate we developed GOXG1 and GOXG2. These conjugates bear a carboxylate ester linker directly attached to the primary and the secondary alcohol group of the drug respectively, followed by oxime and amide bond. The primary alcohol of gemcitabine has been used since it is involved in the phosphorylation process, through which gemcitabine exerts its cytotoxic effect. Therefore, we aimed to block the primary alcohol and examine its effect (GOXG1) and also take advantage of the secondary alcohol which could lead to a PDC with a completely different profile (GOXG2), although they share structural similarities. For a slower drug release rate, we designed and developed the PDC GN4OXG that contains an amide bond on the 4-N position of the parent drug followed by click oxime ligation and another amide bond. The stability of this molecule should be enhanced since it is devoid of rapidly hydrolyzable ester bonds. Furthermore, in this PDC since the 4-NH2 moiety of gemcitabine is capped it could further surmount the rapid gemcitabine metabolism that leads to the formation of dFdU, after the enzymatic 4-N deamination of gemcitabine by cytidine deaminase (CDA).

   Click to Show/Hide
Description
This experiment showed that the more labile pro-drug was GOXG2, which was almost fully degraded in less than 10 hours while GN4OXG and GOXG1 showed similar, enhanced stability compared to GOXG2.
In Vitro Model Prostate carcinoma DU145 cell CVCL_0105
Experiment 2 Reporting the Activity Data of This PDC [29]
Indication Prostate cancer
Efficacy Data Half maximal inhibitory concentration (IC50) 494 ± 93 nM
MOA of PDC
We aimed to construct PDCs with linker controllable drug release rates simply by manipulating the linker unit. For a more rapid drug-release rate we developed GOXG1 and GOXG2. These conjugates bear a carboxylate ester linker directly attached to the primary and the secondary alcohol group of the drug respectively, followed by oxime and amide bond. The primary alcohol of gemcitabine has been used since it is involved in the phosphorylation process, through which gemcitabine exerts its cytotoxic effect. Therefore, we aimed to block the primary alcohol and examine its effect (GOXG1) and also take advantage of the secondary alcohol which could lead to a PDC with a completely different profile (GOXG2), although they share structural similarities. For a slower drug release rate, we designed and developed the PDC GN4OXG that contains an amide bond on the 4-N position of the parent drug followed by click oxime ligation and another amide bond. The stability of this molecule should be enhanced since it is devoid of rapidly hydrolyzable ester bonds. Furthermore, in this PDC since the 4-NH2 moiety of gemcitabine is capped it could further surmount the rapid gemcitabine metabolism that leads to the formation of dFdU, after the enzymatic 4-N deamination of gemcitabine by cytidine deaminase (CDA).

   Click to Show/Hide
Description
The most potent among them was found to be pro-drug GOXG2 with IC50 494 ± 93 nM and 675 ± 82 nM against DU145 and PC3, respectively. Regarding the other two conjugates, GN4OXG was found to be the next most cytotoxic compound against DU145 with IC50 590 ± 62 nM, followed by the least toxic GOXG1 with IC50 611 ± 80 nM. In contrast, GOXG1 was the second more toxic against PC3 with IC50 754 ± 142 nM, followed by the least toxic GN4OXG which showed IC50 833 ± 27 nM. The results are summarized in Table 2.

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In Vitro Model Prostate carcinoma DU145 cell CVCL_0105
Experiment 3 Reporting the Activity Data of This PDC [29]
Indication Prostate cancer
Efficacy Data Half maximal inhibitory concentration (IC50) 675 ± 82 nM
MOA of PDC
We aimed to construct PDCs with linker controllable drug release rates simply by manipulating the linker unit. For a more rapid drug-release rate we developed GOXG1 and GOXG2. These conjugates bear a carboxylate ester linker directly attached to the primary and the secondary alcohol group of the drug respectively, followed by oxime and amide bond. The primary alcohol of gemcitabine has been used since it is involved in the phosphorylation process, through which gemcitabine exerts its cytotoxic effect. Therefore, we aimed to block the primary alcohol and examine its effect (GOXG1) and also take advantage of the secondary alcohol which could lead to a PDC with a completely different profile (GOXG2), although they share structural similarities. For a slower drug release rate, we designed and developed the PDC GN4OXG that contains an amide bond on the 4-N position of the parent drug followed by click oxime ligation and another amide bond. The stability of this molecule should be enhanced since it is devoid of rapidly hydrolyzable ester bonds. Furthermore, in this PDC since the 4-NH2 moiety of gemcitabine is capped it could further surmount the rapid gemcitabine metabolism that leads to the formation of dFdU, after the enzymatic 4-N deamination of gemcitabine by cytidine deaminase (CDA).

   Click to Show/Hide
Description
The most potent among them was found to be pro-drug GOXG2 with IC50 494 ± 93 nM and 675 ± 82 nM against DU145 and PC3, respectively. Regarding the other two conjugates, GN4OXG was found to be the next most cytotoxic compound against DU145 with IC50 590 ± 62 nM, followed by the least toxic GOXG1 with IC50 611 ± 80 nM. In contrast, GOXG1 was the second more toxic against PC3 with IC50 754 ± 142 nM, followed by the least toxic GN4OXG which showed IC50 833 ± 27 nM. The results are summarized in Table 2.

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In Vitro Model Prostate carcinoma PC-3 cell CVCL_0035
GOXG1 [Investigative]
 PDC Info 
Identified from the Human Clinical Data
Click To Hide/Show 1 Activity Data Related to This Level
Experiment 1 Reporting the Activity Data of This PDC [29]
Indication Prostate cancer
Efficacy Data Relative intensity 30%
Administration Time 60 min
Administration Dosage 1 μg/ml
MOA of PDC
We aimed to construct PDCs with linker controllable drug release rates simply by manipulating the linker unit. For a more rapid drug-release rate we developed GOXG1 and GOXG2. These conjugates bear a carboxylate ester linker directly attached to the primary and the secondary alcohol group of the drug respectively, followed by oxime and amide bond. The primary alcohol of gemcitabine has been used since it is involved in the phosphorylation process, through which gemcitabine exerts its cytotoxic effect. Therefore, we aimed to block the primary alcohol and examine its effect (GOXG1) and also take advantage of the secondary alcohol which could lead to a PDC with a completely different profile (GOXG2), although they share structural similarities. For a slower drug release rate, we designed and developed the PDC GN4OXG that contains an amide bond on the 4-N position of the parent drug followed by click oxime ligation and another amide bond. The stability of this molecule should be enhanced since it is devoid of rapidly hydrolyzable ester bonds. Furthermore, in this PDC since the 4-NH2 moiety of gemcitabine is capped it could further surmount the rapid gemcitabine metabolism that leads to the formation of dFdU, after the enzymatic 4-N deamination of gemcitabine by cytidine deaminase (CDA).

   Click to Show/Hide
Description
GOXG2 was the least stable, followed by GOXG1, while GN4OXG was the most stable.
In Vivo Model Human plasma.
Revealed Based on the Cell Line Data
Click To Hide/Show 3 Activity Data Related to This Level
Experiment 1 Reporting the Activity Data of This PDC [29]
Indication Prostate cancer
Efficacy Data Relative intensity 10%
Administration Time 10 h
Administration Dosage 1 μM
MOA of PDC
We aimed to construct PDCs with linker controllable drug release rates simply by manipulating the linker unit. For a more rapid drug-release rate we developed GOXG1 and GOXG2. These conjugates bear a carboxylate ester linker directly attached to the primary and the secondary alcohol group of the drug respectively, followed by oxime and amide bond. The primary alcohol of gemcitabine has been used since it is involved in the phosphorylation process, through which gemcitabine exerts its cytotoxic effect. Therefore, we aimed to block the primary alcohol and examine its effect (GOXG1) and also take advantage of the secondary alcohol which could lead to a PDC with a completely different profile (GOXG2), although they share structural similarities. For a slower drug release rate, we designed and developed the PDC GN4OXG that contains an amide bond on the 4-N position of the parent drug followed by click oxime ligation and another amide bond. The stability of this molecule should be enhanced since it is devoid of rapidly hydrolyzable ester bonds. Furthermore, in this PDC since the 4-NH2 moiety of gemcitabine is capped it could further surmount the rapid gemcitabine metabolism that leads to the formation of dFdU, after the enzymatic 4-N deamination of gemcitabine by cytidine deaminase (CDA).

   Click to Show/Hide
Description
This experiment showed that the more labile pro-drug was GOXG2, which was almost fully degraded in less than 10 hours while GN4OXG and GOXG1 showed similar, enhanced stability compared to GOXG2.
In Vitro Model Prostate carcinoma DU145 cell CVCL_0105
Experiment 2 Reporting the Activity Data of This PDC [29]
Indication Prostate cancer
Efficacy Data Half maximal inhibitory concentration (IC50) 611 ± 80 nM
MOA of PDC
We aimed to construct PDCs with linker controllable drug release rates simply by manipulating the linker unit. For a more rapid drug-release rate we developed GOXG1 and GOXG2. These conjugates bear a carboxylate ester linker directly attached to the primary and the secondary alcohol group of the drug respectively, followed by oxime and amide bond. The primary alcohol of gemcitabine has been used since it is involved in the phosphorylation process, through which gemcitabine exerts its cytotoxic effect. Therefore, we aimed to block the primary alcohol and examine its effect (GOXG1) and also take advantage of the secondary alcohol which could lead to a PDC with a completely different profile (GOXG2), although they share structural similarities. For a slower drug release rate, we designed and developed the PDC GN4OXG that contains an amide bond on the 4-N position of the parent drug followed by click oxime ligation and another amide bond. The stability of this molecule should be enhanced since it is devoid of rapidly hydrolyzable ester bonds. Furthermore, in this PDC since the 4-NH2 moiety of gemcitabine is capped it could further surmount the rapid gemcitabine metabolism that leads to the formation of dFdU, after the enzymatic 4-N deamination of gemcitabine by cytidine deaminase (CDA).

   Click to Show/Hide
Description
The most potent among them was found to be pro-drug GOXG2 with IC50 494 ± 93 nM and 675 ± 82 nM against DU145 and PC3, respectively. Regarding the other two conjugates, GN4OXG was found to be the next most cytotoxic compound against DU145 with IC50 590 ± 62 nM, followed by the least toxic GOXG1 with IC50 611 ± 80 nM. In contrast, GOXG1 was the second more toxic against PC3 with IC50 754 ± 142 nM, followed by the least toxic GN4OXG which showed IC50 833 ± 27 nM. The results are summarized in Table 2.

   Click to Show/Hide
In Vitro Model Prostate carcinoma DU145 cell CVCL_0105
Experiment 3 Reporting the Activity Data of This PDC [29]
Indication Prostate cancer
Efficacy Data Half maximal inhibitory concentration (IC50) 754 ± 142 nM
MOA of PDC
We aimed to construct PDCs with linker controllable drug release rates simply by manipulating the linker unit. For a more rapid drug-release rate we developed GOXG1 and GOXG2. These conjugates bear a carboxylate ester linker directly attached to the primary and the secondary alcohol group of the drug respectively, followed by oxime and amide bond. The primary alcohol of gemcitabine has been used since it is involved in the phosphorylation process, through which gemcitabine exerts its cytotoxic effect. Therefore, we aimed to block the primary alcohol and examine its effect (GOXG1) and also take advantage of the secondary alcohol which could lead to a PDC with a completely different profile (GOXG2), although they share structural similarities. For a slower drug release rate, we designed and developed the PDC GN4OXG that contains an amide bond on the 4-N position of the parent drug followed by click oxime ligation and another amide bond. The stability of this molecule should be enhanced since it is devoid of rapidly hydrolyzable ester bonds. Furthermore, in this PDC since the 4-NH2 moiety of gemcitabine is capped it could further surmount the rapid gemcitabine metabolism that leads to the formation of dFdU, after the enzymatic 4-N deamination of gemcitabine by cytidine deaminase (CDA).

   Click to Show/Hide
Description
The most potent among them was found to be pro-drug GOXG2 with IC50 494 ± 93 nM and 675 ± 82 nM against DU145 and PC3, respectively. Regarding the other two conjugates, GN4OXG was found to be the next most cytotoxic compound against DU145 with IC50 590 ± 62 nM, followed by the least toxic GOXG1 with IC50 611 ± 80 nM. In contrast, GOXG1 was the second more toxic against PC3 with IC50 754 ± 142 nM, followed by the least toxic GN4OXG which showed IC50 833 ± 27 nM. The results are summarized in Table 2.

   Click to Show/Hide
In Vitro Model Prostate carcinoma PC-3 cell CVCL_0035
GN4OXG [Investigative]
 PDC Info 
Identified from the Human Clinical Data
Click To Hide/Show 1 Activity Data Related to This Level
Experiment 1 Reporting the Activity Data of This PDC [29]
Indication Prostate cancer
Efficacy Data Relative intensity 75%
Administration Time 60 min
Administration Dosage 1 μg/ml
MOA of PDC
We aimed to construct PDCs with linker controllable drug release rates simply by manipulating the linker unit. For a more rapid drug-release rate we developed GOXG1 and GOXG2. These conjugates bear a carboxylate ester linker directly attached to the primary and the secondary alcohol group of the drug respectively, followed by oxime and amide bond. The primary alcohol of gemcitabine has been used since it is involved in the phosphorylation process, through which gemcitabine exerts its cytotoxic effect. Therefore, we aimed to block the primary alcohol and examine its effect (GOXG1) and also take advantage of the secondary alcohol which could lead to a PDC with a completely different profile (GOXG2), although they share structural similarities. For a slower drug release rate, we designed and developed the PDC GN4OXG that contains an amide bond on the 4-N position of the parent drug followed by click oxime ligation and another amide bond. The stability of this molecule should be enhanced since it is devoid of rapidly hydrolyzable ester bonds. Furthermore, in this PDC since the 4-NH2 moiety of gemcitabine is capped it could further surmount the rapid gemcitabine metabolism that leads to the formation of dFdU, after the enzymatic 4-N deamination of gemcitabine by cytidine deaminase (CDA).

   Click to Show/Hide
Description
GOXG2 was the least stable, followed by GOXG1, while GN4OXG was the most stable.
In Vivo Model Human plasma.
Revealed Based on the Cell Line Data
Click To Hide/Show 3 Activity Data Related to This Level
Experiment 1 Reporting the Activity Data of This PDC [29]
Indication Prostate cancer
Efficacy Data Relative intensity 12%
Administration Time 10 h
Administration Dosage 1 μM
MOA of PDC
We aimed to construct PDCs with linker controllable drug release rates simply by manipulating the linker unit. For a more rapid drug-release rate we developed GOXG1 and GOXG2. These conjugates bear a carboxylate ester linker directly attached to the primary and the secondary alcohol group of the drug respectively, followed by oxime and amide bond. The primary alcohol of gemcitabine has been used since it is involved in the phosphorylation process, through which gemcitabine exerts its cytotoxic effect. Therefore, we aimed to block the primary alcohol and examine its effect (GOXG1) and also take advantage of the secondary alcohol which could lead to a PDC with a completely different profile (GOXG2), although they share structural similarities. For a slower drug release rate, we designed and developed the PDC GN4OXG that contains an amide bond on the 4-N position of the parent drug followed by click oxime ligation and another amide bond. The stability of this molecule should be enhanced since it is devoid of rapidly hydrolyzable ester bonds. Furthermore, in this PDC since the 4-NH2 moiety of gemcitabine is capped it could further surmount the rapid gemcitabine metabolism that leads to the formation of dFdU, after the enzymatic 4-N deamination of gemcitabine by cytidine deaminase (CDA).

   Click to Show/Hide
Description
This experiment showed that the more labile pro-drug was GOXG2, which was almost fully degraded in less than 10 hours while GN4OXG and GOXG1 showed similar, enhanced stability compared to GOXG2.
In Vitro Model Prostate carcinoma DU145 cell CVCL_0105
Experiment 2 Reporting the Activity Data of This PDC [29]
Indication Prostate cancer
Efficacy Data Half maximal inhibitory concentration (IC50) 590 ± 62 nM
MOA of PDC
We aimed to construct PDCs with linker controllable drug release rates simply by manipulating the linker unit. For a more rapid drug-release rate we developed GOXG1 and GOXG2. These conjugates bear a carboxylate ester linker directly attached to the primary and the secondary alcohol group of the drug respectively, followed by oxime and amide bond. The primary alcohol of gemcitabine has been used since it is involved in the phosphorylation process, through which gemcitabine exerts its cytotoxic effect. Therefore, we aimed to block the primary alcohol and examine its effect (GOXG1) and also take advantage of the secondary alcohol which could lead to a PDC with a completely different profile (GOXG2), although they share structural similarities. For a slower drug release rate, we designed and developed the PDC GN4OXG that contains an amide bond on the 4-N position of the parent drug followed by click oxime ligation and another amide bond. The stability of this molecule should be enhanced since it is devoid of rapidly hydrolyzable ester bonds. Furthermore, in this PDC since the 4-NH2 moiety of gemcitabine is capped it could further surmount the rapid gemcitabine metabolism that leads to the formation of dFdU, after the enzymatic 4-N deamination of gemcitabine by cytidine deaminase (CDA).

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Description
The most potent among them was found to be pro-drug GOXG2 with IC50 494 ± 93 nM and 675 ± 82 nM against DU145 and PC3, respectively. Regarding the other two conjugates, GN4OXG was found to be the next most cytotoxic compound against DU145 with IC50 590 ± 62 nM, followed by the least toxic GOXG1 with IC50 611 ± 80 nM. In contrast, GOXG1 was the second more toxic against PC3 with IC50 754 ± 142 nM, followed by the least toxic GN4OXG which showed IC50 833 ± 27 nM. The results are summarized in Table 2.

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In Vitro Model Prostate carcinoma DU145 cell CVCL_0105
Experiment 3 Reporting the Activity Data of This PDC [29]
Indication Prostate cancer
Efficacy Data Half maximal inhibitory concentration (IC50) 833 ± 27 nM
MOA of PDC
We aimed to construct PDCs with linker controllable drug release rates simply by manipulating the linker unit. For a more rapid drug-release rate we developed GOXG1 and GOXG2. These conjugates bear a carboxylate ester linker directly attached to the primary and the secondary alcohol group of the drug respectively, followed by oxime and amide bond. The primary alcohol of gemcitabine has been used since it is involved in the phosphorylation process, through which gemcitabine exerts its cytotoxic effect. Therefore, we aimed to block the primary alcohol and examine its effect (GOXG1) and also take advantage of the secondary alcohol which could lead to a PDC with a completely different profile (GOXG2), although they share structural similarities. For a slower drug release rate, we designed and developed the PDC GN4OXG that contains an amide bond on the 4-N position of the parent drug followed by click oxime ligation and another amide bond. The stability of this molecule should be enhanced since it is devoid of rapidly hydrolyzable ester bonds. Furthermore, in this PDC since the 4-NH2 moiety of gemcitabine is capped it could further surmount the rapid gemcitabine metabolism that leads to the formation of dFdU, after the enzymatic 4-N deamination of gemcitabine by cytidine deaminase (CDA).

   Click to Show/Hide
Description
The most potent among them was found to be pro-drug GOXG2 with IC50 494 ± 93 nM and 675 ± 82 nM against DU145 and PC3, respectively. Regarding the other two conjugates, GN4OXG was found to be the next most cytotoxic compound against DU145 with IC50 590 ± 62 nM, followed by the least toxic GOXG1 with IC50 611 ± 80 nM. In contrast, GOXG1 was the second more toxic against PC3 with IC50 754 ± 142 nM, followed by the least toxic GN4OXG which showed IC50 833 ± 27 nM. The results are summarized in Table 2.

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In Vitro Model Prostate carcinoma PC-3 cell CVCL_0035
D-Lys6-GnRH-gemcitabine(2G2) [Investigative]
 PDC Info 
Revealed Based on the Cell Line Data
Click To Hide/Show 5 Activity Data Related to This Level
Experiment 1 Reporting the Activity Data of This PDC [30]
Indication Tumor
Efficacy Data Half Maximal Inhibitory Concentration (IC50) 0.11 nM
Description
The presented data show that 2G2, 2G1 and GSHG bind to GnRH-R with 95.5-, 15.2-, and 4.4-fold higher affinity, respectively, than that of the native peptide D-Lys6-GnRH (10.5 ± 0.2 nM, according to our former study [3]).
In Vitro Model Normal HEK293 cell CVCL_0045
Experiment 2 Reporting the Activity Data of This PDC [30]
Indication Tumor
Efficacy Data Half Maximal Inhibitory Concentration (IC50) 449.1 nM
Description
GSHGpossesses the highest cytotoxic effect among the three conjugates, which is comparable with that of gemcitabine in the examined cell lines and especially regarding MCF-7 cells.
In Vitro Model Invasive breast carcinoma MCF-7 cell CVCL_0031
Experiment 3 Reporting the Activity Data of This PDC [30]
Indication Tumor
Efficacy Data Half Maximal Inhibitory Concentration (IC50) 9761 nM
Description
GSHGpossesses the highest cytotoxic effect among the three conjugates, which is comparable with that of gemcitabine in the examined cell lines and especially regarding MCF-7 cells.
In Vitro Model Prostate carcinoma DU145 cell CVCL_0105
Experiment 4 Reporting the Activity Data of This PDC [30]
Indication Tumor
Efficacy Data Half Maximal Inhibitory Concentration (IC50) > 40000 nM
Description
GSHGpossesses the highest cytotoxic effect among the three conjugates, which is comparable with that of gemcitabine in the examined cell lines and especially regarding MCF-7 cells.
In Vitro Model Prostate carcinoma PC-3 cell CVCL_0035
Experiment 5 Reporting the Activity Data of This PDC [30]
Indication Tumor
Efficacy Data Half Maximal Inhibitory Concentration (IC50) > 40000 nM
Description
GSHGpossesses the highest cytotoxic effect among the three conjugates, which is comparable with that of gemcitabine in the examined cell lines and especially regarding MCF-7 cells.
In Vitro Model Breast adenocarcinoma MDA-MB-231 cell CVCL_0062
D-Lys6-GnRH-gemcitabine(2G1) [Investigative]
 PDC Info 
Revealed Based on the Cell Line Data
Click To Hide/Show 5 Activity Data Related to This Level
Experiment 1 Reporting the Activity Data of This PDC [30]
Indication Tumor
Efficacy Data Half Maximal Inhibitory Concentration (IC50) 0.69 nM
Description
The presented data show that 2G2, 2G1 and GSHG bind to GnRH-R with 95.5-, 15.2-, and 4.4-fold higher affinity, respectively, than that of the native peptide D-Lys6-GnRH (10.5 ± 0.2 nM, according to our former study [3]).
In Vitro Model Normal HEK293 cell CVCL_0045
Experiment 2 Reporting the Activity Data of This PDC [30]
Indication Tumor
Efficacy Data Half Maximal Inhibitory Concentration (IC50) 621.3 nM
Description
GSHGpossesses the highest cytotoxic effect among the three conjugates, which is comparable with that of gemcitabine in the examined cell lines and especially regarding MCF-7 cells.
In Vitro Model Invasive breast carcinoma MCF-7 cell CVCL_0031
Experiment 3 Reporting the Activity Data of This PDC [30]
Indication Tumor
Efficacy Data Half Maximal Inhibitory Concentration (IC50) > 40000 nM
Description
GSHGpossesses the highest cytotoxic effect among the three conjugates, which is comparable with that of gemcitabine in the examined cell lines and especially regarding MCF-7 cells.
In Vitro Model Prostate carcinoma DU145 cell CVCL_0105
Experiment 4 Reporting the Activity Data of This PDC [30]
Indication Tumor
Efficacy Data Half Maximal Inhibitory Concentration (IC50) > 40000 nM
Description
GSHGpossesses the highest cytotoxic effect among the three conjugates, which is comparable with that of gemcitabine in the examined cell lines and especially regarding MCF-7 cells.
In Vitro Model Prostate carcinoma PC-3 cell CVCL_0035
Experiment 5 Reporting the Activity Data of This PDC [30]
Indication Tumor
Efficacy Data Half Maximal Inhibitory Concentration (IC50) > 40000 nM
Description
GSHGpossesses the highest cytotoxic effect among the three conjugates, which is comparable with that of gemcitabine in the examined cell lines and especially regarding MCF-7 cells.
In Vitro Model Breast adenocarcinoma MDA-MB-231 cell CVCL_0062
EETI-2.5Z-Val-Ala-PAB-gemcitabine [Investigative]
 PDC Info 
Revealed Based on the Cell Line Data
Click To Hide/Show 7 Activity Data Related to This Level
Experiment 1 Reporting the Activity Data of This PDC [31]
Indication Glioblastoma
Efficacy Data Half Maximal Inhibitory Concentration (IC50) 1.5 ± 0.2 nM
Evaluation Method CCK-8 assay
Description
Cell proliferation was quantified 4 d after treatment with each compoundusing CCK-8 colorimetric assays and compared to the untreated control.
In Vitro Model Glioblastoma U-87MG cell CVCL_0022
Experiment 2 Reporting the Activity Data of This PDC [31]
Indication Breast cancer
Efficacy Data Half Maximal Effective Concentration (EC50) 0.6 ± 0.1 nM
Evaluation Method CCK-8 assay
Description
Cell proliferation was quantified 4 d after treatment with 5 d or 3 using CCK-8 colorimetric assays and compared to the untreated control. Metabolic activity measured by CCK-8 was validated by quantifying celldeath using Trypan Blue.
In Vitro Model Breast adenocarcinoma MDA-MB-468 cell CVCL_0419
Experiment 3 Reporting the Activity Data of This PDC [31]
Indication Pancreatic cancer
Efficacy Data Half Maximal Effective Concentration (EC50) 1.8 ± 0.8 nM
Evaluation Method CCK-8 assay
Description
Cell proliferation was quantified 4 d after treatment with 5 d or 3 using CCK-8 colorimetric assays and compared to the untreated control. Metabolic activity measured by CCK-8 was validated by quantifying celldeath using Trypan Blue.
In Vitro Model Pancreatic ductal adenocarcinoma BxPC-3 cell CVCL_0186
Experiment 4 Reporting the Activity Data of This PDC [31]
Indication Pancreatic cancer
Efficacy Data Half Maximal Effective Concentration (EC50) 2.1 ± 0.2 nM
Evaluation Method CCK-8 assay
Description
Cell proliferation was quantified 4 d after treatment with 5 d or 3 using CCK-8 colorimetric assays and compared to the untreated control. Metabolic activity measured by CCK-8 was validated by quantifying celldeath using Trypan Blue.
In Vitro Model Pancreatic ductal adenocarcinoma PANC-1 cell CVCL_0480
Experiment 5 Reporting the Activity Data of This PDC [31]
Indication Ovarian cancer
Efficacy Data Half Maximal Effective Concentration (EC50) 2.3 ± 0.5 nM
Evaluation Method CCK-8 assay
Description
Cell proliferation was quantified 4 d after treatment with 5 d or 3 using CCK-8 colorimetric assays and compared to the untreated control. Metabolic activity measured by CCK-8 was validated by quantifying celldeath using Trypan Blue.
In Vitro Model Ovarian endometrioid adenocarcinoma A2780 cell CVCL_0134
Experiment 6 Reporting the Activity Data of This PDC [31]
Indication Glioblastoma
Efficacy Data Half Maximal Effective Concentration (EC50) 7.9 ± 0.8 nM
Evaluation Method CCK-8 assay
Description
Cell proliferation was quantified 4 d after treatment with 5 d or 3 using CCK-8 colorimetric assays and compared to the untreated control. Metabolic activity measured by CCK-8 was validated by quantifying celldeath using Trypan Blue.
In Vitro Model Glioblastoma D-270MG cell CVCL_S751
Experiment 7 Reporting the Activity Data of This PDC [31]
Indication Glioblastoma
Efficacy Data Half Maximal Effective Concentration (EC50) 9.0 ± 1.8 nM
Evaluation Method CCK-8 assay
Description
Cell proliferation was quantified 4 d after treatment with 5 d or 3 using CCK-8 colorimetric assays and compared to the untreated control. Metabolic activity measured by CCK-8 was validated by quantifying celldeath using Trypan Blue.
In Vitro Model Glioblastoma U-87MG cell CVCL_0022
D-Lys6-GnRH-gemcitabine(GSHG) [Investigative]
 PDC Info 
Revealed Based on the Cell Line Data
Click To Hide/Show 5 Activity Data Related to This Level
Experiment 1 Reporting the Activity Data of This PDC [30]
Indication Tumor
Efficacy Data Half Maximal Inhibitory Concentration (IC50) 2.40 nM
Description
The presented data show that 2G2, 2G1 and GSHG bind to GnRH-R with 95.5-, 15.2-, and 4.4-fold higher affinity, respectively, than that of the native peptide D-Lys6-GnRH (10.5 ± 0.2 nM, according to our former study [3]).
In Vitro Model Normal HEK293 cell CVCL_0045
Experiment 2 Reporting the Activity Data of This PDC [30]
Indication Tumor
Efficacy Data Half Maximal Inhibitory Concentration (IC50) 55.5 nM
Description
GSHGpossesses the highest cytotoxic effect among the three conjugates, which is comparable with that of gemcitabine in the examined cell lines and especially regarding MCF-7 cells.
In Vitro Model Invasive breast carcinoma MCF-7 cell CVCL_0031
Experiment 3 Reporting the Activity Data of This PDC [30]
Indication Tumor
Efficacy Data Half Maximal Inhibitory Concentration (IC50) 684 nM
Description
GSHGpossesses the highest cytotoxic effect among the three conjugates, which is comparable with that of gemcitabine in the examined cell lines and especially regarding MCF-7 cells.
In Vitro Model Prostate carcinoma DU145 cell CVCL_0105
Experiment 4 Reporting the Activity Data of This PDC [30]
Indication Tumor
Efficacy Data Half Maximal Inhibitory Concentration (IC50) 937 nM
Description
GSHGpossesses the highest cytotoxic effect among the three conjugates, which is comparable with that of gemcitabine in the examined cell lines and especially regarding MCF-7 cells.
In Vitro Model Prostate carcinoma PC-3 cell CVCL_0035
Experiment 5 Reporting the Activity Data of This PDC [30]
Indication Tumor
Efficacy Data Half Maximal Inhibitory Concentration (IC50) 2387 nM
Description
GSHGpossesses the highest cytotoxic effect among the three conjugates, which is comparable with that of gemcitabine in the examined cell lines and especially regarding MCF-7 cells.
In Vitro Model Breast adenocarcinoma MDA-MB-231 cell CVCL_0062
EETI-2.5Z-amide -gemcitabine [Investigative]
 PDC Info 
Revealed Based on the Cell Line Data
Click To Hide/Show 2 Activity Data Related to This Level
Experiment 1 Reporting the Activity Data of This PDC [31]
Indication Glioblastoma
Efficacy Data Half Maximal Inhibitory Concentration (IC50) 2.8 ± 0.2 nM
Evaluation Method CCK-8 assay
Description
Cell proliferation was quantified 4 d after treatment with each compoundusing CCK-8 colorimetric assays and compared to the untreated control.
In Vitro Model Glioblastoma U-87MG cell CVCL_0022
Experiment 2 Reporting the Activity Data of This PDC [31]
Indication Glioblastoma
Efficacy Data Half Maximal Effective Concentration (EC50) 8.9 ± 1.2 nM
Evaluation Method CCK-8 assay
Description
Cell proliferation was quantified 4 d after treatment with each compoundusing CCK-8 colorimetric assays and compared to the untreated control.
In Vitro Model Glioblastoma U-87MG cell CVCL_0022
EETI-2.5Z-carbamate -gemcitabine [Investigative]
 PDC Info 
Revealed Based on the Cell Line Data
Click To Hide/Show 2 Activity Data Related to This Level
Experiment 1 Reporting the Activity Data of This PDC [31]
Indication Glioblastoma
Efficacy Data Half Maximal Inhibitory Concentration (IC50) 3.3 ± 0.2 nM
Evaluation Method CCK-8 assay
Description
Cell proliferation was quantified 4 d after treatment with each compoundusing CCK-8 colorimetric assays and compared to the untreated control.
In Vitro Model Glioblastoma U-87MG cell CVCL_0022
Experiment 2 Reporting the Activity Data of This PDC [31]
Indication Glioblastoma
Efficacy Data Half Maximal Effective Concentration (EC50) > 1000 nM
Evaluation Method CCK-8 assay
Description
Cell proliferation was quantified 4 d after treatment with each compoundusing CCK-8 colorimetric assays and compared to the untreated control.
In Vitro Model Glioblastoma U-87MG cell CVCL_0022
EETI-2.5Z-ester-gemcitabine [Investigative]
 PDC Info 
Revealed Based on the Cell Line Data
Click To Hide/Show 2 Activity Data Related to This Level
Experiment 1 Reporting the Activity Data of This PDC [31]
Indication Glioblastoma
Efficacy Data Half Maximal Inhibitory Concentration (IC50) 5.2 ± 3.6 nM
Evaluation Method CCK-8 assay
Description
Cell proliferation was quantified 4 d after treatment with each compoundusing CCK-8 colorimetric assays and compared to the untreated control.
In Vitro Model Glioblastoma U-87MG cell CVCL_0022
Experiment 2 Reporting the Activity Data of This PDC [31]
Indication Glioblastoma
Efficacy Data Half Maximal Effective Concentration (EC50) 8.5 ± 3.3 nM
Evaluation Method CCK-8 assay
Description
Cell proliferation was quantified 4 d after treatment with each compoundusing CCK-8 colorimetric assays and compared to the untreated control.
In Vitro Model Glioblastoma U-87MG cell CVCL_0022
OGF-Gem [Investigative]
 PDC Info 
Revealed Based on the Cell Line Data
Click To Hide/Show 2 Activity Data Related to This Level
Experiment 1 Reporting the Activity Data of This PDC [32]
Indication Pancreatic ductal adenocarcinoma
Efficacy Data Half Maximal Inhibitory Concentration (IC50) 17.63 ± 2.334 nM
Evaluation Method MTT assay
Administration Time 72 h
MOA of PDC
Therefore, we designed, synthesized, and characterized an OGF-Gem conjugate, where OGF and Gem are tethered by an organic linker (Figure 1). Gem was subjected to selective protection using the tert-butoxycarbonyl (Boc) group and prepared as gemcitabine hemisuccinate. 5-O-diBoc-gemcitabine hemisuccinate was conjugated with the OGF peptide in solution. We demonstrated the cytotoxic activity of the OGF-Gem conjugate against pancreatic cancer cell lines, including the metastatic line (MIA PaCa-2 and AsPC-1). Furthermore, we confirmed that OGF-Gem is either not cytotoxic or significantly less cytotoxic to two non-tumor-transformed human cellskidney (HEK-293) and skin fibroblast cells (HDFa). We also determined the effect of OGF-Gem on cell cycle inhibition, and the inhibition of cell proliferation, senescent cells, and apoptosis. We have demonstrated that OGF-Gem has antimetastatic potential due to inhibited pancreatic tumor cell (AsPC-1)-induced platelet aggregation. This can significantly impact the inhibition of disease progression (metastasis) of pancreatic cancer.

   Click to Show/Hide
Description
The tested compounds cytotoxic activity was determined using the MTT test, which is based on the ability to convert tetrazole salts to water insoluble formazan through mitochondrial dehydrogenases. Our results show a high cytotoxic effect on all pancreatic cancer cell lines. Exposing pancreatic cell lines MIA PaCa-2 and AsPC-1 to OGF-Gem decreased viability. Importantly, the OGF-Gem conjugate demonstrated a more pronounced cytotoxic effect against the metastatic pancreatic cancer cell line AsPC-1 compared to the commonly used chemotherapeutic agent. The results obtained for non-tumor-transformed cellsa human embryonic kidney line HEK-293 and human primary dermal fibroblast line HDFa presented a slight cytotoxicity effect from the OGF-Gem derivative within 3 days of incubation for all tested concentrations. Interestingly, an 80% reduction in HEK-293 cell viability was observed for the 100 nM Gem compared to the 100 nM OGF-Gem derivative. In HDFa cells, 100 nM Gem reduced viability to 35%, while the OGF-Gem conjugate slightly decreased the viability (to 75% viability) after 72 h of incubation. Based on the analysis of the results, concentrations of 3.125, 12.5, 50, and 100 nM, as well as an incubation time of 72 h, were selected for further experiments on the three pancreatic cancer cell lines.

   Click to Show/Hide
In Vitro Model Pancreatic ductal adenocarcinoma MIA PaCa-2 cell CVCL_0428
Experiment 2 Reporting the Activity Data of This PDC [32]
Indication Pancreatic ductal adenocarcinoma
Efficacy Data Half Maximal Inhibitory Concentration (IC50) 27.44 ± 9.161 nM
Evaluation Method MTT assay
Administration Time 72 h
MOA of PDC
Therefore, we designed, synthesized, and characterized an OGF-Gem conjugate, where OGF and Gem are tethered by an organic linker (Figure 1). Gem was subjected to selective protection using the tert-butoxycarbonyl (Boc) group and prepared as gemcitabine hemisuccinate. 5-O-diBoc-gemcitabine hemisuccinate was conjugated with the OGF peptide in solution. We demonstrated the cytotoxic activity of the OGF-Gem conjugate against pancreatic cancer cell lines, including the metastatic line (MIA PaCa-2 and AsPC-1). Furthermore, we confirmed that OGF-Gem is either not cytotoxic or significantly less cytotoxic to two non-tumor-transformed human cellskidney (HEK-293) and skin fibroblast cells (HDFa). We also determined the effect of OGF-Gem on cell cycle inhibition, and the inhibition of cell proliferation, senescent cells, and apoptosis. We have demonstrated that OGF-Gem has antimetastatic potential due to inhibited pancreatic tumor cell (AsPC-1)-induced platelet aggregation. This can significantly impact the inhibition of disease progression (metastasis) of pancreatic cancer.

   Click to Show/Hide
Description
The tested compounds cytotoxic activity was determined using the MTT test, which is based on the ability to convert tetrazole salts to water insoluble formazan through mitochondrial dehydrogenases. Our results show a high cytotoxic effect on all pancreatic cancer cell lines. Exposing pancreatic cell lines MIA PaCa-2 and AsPC-1 to OGF-Gem decreased viability. Importantly, the OGF-Gem conjugate demonstrated a more pronounced cytotoxic effect against the metastatic pancreatic cancer cell line AsPC-1 compared to the commonly used chemotherapeutic agent. The results obtained for non-tumor-transformed cellsa human embryonic kidney line HEK-293 and human primary dermal fibroblast line HDFa presented a slight cytotoxicity effect from the OGF-Gem derivative within 3 days of incubation for all tested concentrations. Interestingly, an 80% reduction in HEK-293 cell viability was observed for the 100 nM Gem compared to the 100 nM OGF-Gem derivative. In HDFa cells, 100 nM Gem reduced viability to 35%, while the OGF-Gem conjugate slightly decreased the viability (to 75% viability) after 72 h of incubation. Based on the analysis of the results, concentrations of 3.125, 12.5, 50, and 100 nM, as well as an incubation time of 72 h, were selected for further experiments on the three pancreatic cancer cell lines.

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In Vitro Model Pancreatic ductal adenocarcinoma AsPC-1 cell CVCL_0152
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