DNA synthesis
Nedaplatin (NSC 375101D) targets tumor cell DNA (induces intra- and inter-strand DNA cross-links, IC50=2.8 μM for human lung adenocarcinoma cell line A549; IC50=1.9 μM for human cervical cancer cell line HeLa)[3]
Nedaplatin inhibits DNA topoisomerase II (in vitro enzyme activity inhibition IC50=4.5 μM)[2]

Nedaplatin (Aqupla) is a cisplatin derivative that inhibits tumor colony forming units, with an IC50 value of 28.5 μg/mL. [1] A platinum compound called nedaplatin is used as a cancer chemotherapy drug. The proliferation of SBC-3 cells is inhibited by nedaplatin at concentrations of 0.005 μg/mL, 0.01 μg/mL, 0.025 μg/mL, 0.05 μg/mL, 0.1 μg/mL, 0.25 μg/mL, and 0.5 μg/mL, respectively. Nedaplatin's IC50 value for suppressing SBC-3 cell growth is 0.053 μg/mL.[3]

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Anti-lung cancer cell activity: IC50 values were 2.8 μM for A549, 2.1 μM for PC-9; IC50=5.6 μM for cisplatin-resistant A549/cis cells with a resistance fold of 2, showing weak cross-resistance[3]
- Anti-gastrointestinal cancer cell activity: IC50 values were 3.2 μM for MKN-45, 2.7 μM for SGC-7901 (human gastric cancer cells); IC50=3.5 μM for human colon cancer cell line HCT116[4]
- Anti-gynecological cancer cell activity: IC50 values were 1.9 μM for HeLa, 2.3 μM for SiHa (human cervical cancer cells); IC50=3.1 μM for human ovarian cancer cell line SK-OV-3[5]
- Induces tumor cell apoptosis: Treatment of HeLa cells with 2 μM Nedaplatin for 48 hours resulted in 52% apoptotic cells, accompanied by activation of caspase-3 and caspase-8, and increased PARP cleavage[5]
- Cell cycle arrest: Treatment of A549 cells with 3 μM Nedaplatin for 24 hours increased the proportion of G2/M phase cells from 14% to 42%, blocking DNA synthesis[3]
- DNA cross-linking activity: Incubation with calf thymus DNA resulted in 38% intra-strand cross-link products (5 μM, 24 hours), significantly higher than cisplatin (25%)[2]
- Synergistic effect with combination therapy: Combined treatment of A549 cells with Nedaplatin and paclitaxel (10 nM) reduced IC50 from 2.8 μM to 1.1 μM; combined treatment of MKN-45 cells with Nedaplatin and 5-fluorouracil (10 μM) increased apoptosis rate from 41% to 68%[6]

In comparison to nedaplatin, CDDP, or 5-FU monotherapy, the sequential administration of 5-FU before nedaplatin or CDDP (FN or FC therapy) results in synergistically enhanced inhibition of tumour growth and prolonged survival.[4] When Nedaplatin and Gemcitabine are administered together, the Ma44 tumor model exhibits a synergistically enhanced inhibition of tumor growth. When administered toward the end of the therapy, nedaplatin plus gemcitabine is also effective against Ma44 cells, a model for advanced disease. The NCI-H460 tumor model also demonstrates a strong enhancement of growth inhibition by neoplatin when combined with gemcitabine.[5]

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Human lung cancer xenograft model (BALB/c nude mice): Intravenous injection of Nedaplatin 4 mg/kg once weekly for 3 consecutive weeks achieved a tumor inhibition rate of 65%; the 8 mg/kg dose group had an inhibition rate of 82%, reducing tumor volume from 350 mm³ to 63 mm³[3]
- Human gastric cancer xenograft model (nude mice): Intraperitoneal injection of Nedaplatin 6 mg/kg once every two weeks for 2 consecutive times reduced tumor weight from 1.2 g to 0.3 g, prolonging median survival time from 42 days to 68 days[4]
- Cisplatin-resistant tumor model (A549/cis xenograft nude mice): Intravenous injection of Nedaplatin 8 mg/kg once weekly for 3 consecutive weeks achieved an inhibition rate of 58%, significantly higher than the cisplatin group (22%)[3]
- In vivo efficacy of combination therapy: Nedaplatin (4 mg/kg, intravenous injection, once weekly) combined with paclitaxel (10 mg/kg, intraperitoneal injection, once weekly) for lung cancer xenografts increased tumor inhibition rate from 65% to 89% without significant toxicity increase[6]

DNA topoisomerase II activity inhibition assay: Purified human topoisomerase II was incubated with serial concentrations of Nedaplatin (0.5~20 μM) for 30 minutes, and pBR322 DNA substrate was added. After incubation at 37°C for 1 hour, DNA unwinding and fragmentation were detected by agarose gel electrophoresis. Results showed 50% enzyme activity inhibition at 4.5 μM, and complete blocking of enzyme-mediated DNA topological changes at 10 μM[2]
- DNA cross-linking activity assay: Calf thymus DNA was incubated with 2~10 μM Nedaplatin at 37°C for 24 hours, digested with proteinase K, and cross-link products were detected by HPLC. At 5 μM, intra-strand cross-link products accounted for 38% of total DNA, and inter-strand cross-link products accounted for 8%[2]

An assay for regrowth is used to measure the antitumor activity of the drug treatments, which include the inhibition of cell proliferation (including that of human SCLC cell line SBC-3 and human NSCLC cell line PC-14). To put it briefly, cells are treated with drugs alone or in combination for six days at 37°C in a 100% humidity and 5% CO2 environment. After that, the cells are pipetted six to eight times, or nearly all of them become single cells, and they are counted using a counter. Plotting the percentage of surviving cells (the unaffected cell fraction, fu) against drug concentration yields concentration-effect curves for each drug. To calculate the treated:control cultures' cell proliferation ratio (T:C%), use the formula below: [(the number of treated cells on day 6)/(the number of treated cells on day 0)]/[(the number of control cells on day 6)/(the number of control cells on day 0)] × 100%. The drug concentration needed to cause a 50% decrease in the number of cells is known as the IC50. For each, four or five separate experiments are conducted. In order to assess the impact of the drug treatment schedule on the combination's effect, the cells are treated for three hours either sequentially—first with Nedaplatin and then with irinotecan (Nedaplatin→irinotecan) or simultaneously with both drugs. Cells are exposed to the first drug for three hours during the sequential exposure treatment, then they are immediately exposed to the second drug for three hours after being ished in fresh medium once. Up until assessment, the treated cells are cultivated in a drug-free medium.

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Cell proliferation inhibition assay (MTT method): Different tumor cell lines (A549, HeLa, MKN-45, etc.) were seeded in 96-well plates at 1×10⁴ cells/well, incubated for 24 hours, and treated with serial concentrations of Nedaplatin (0.1~20 μM) for 72 hours. MTT reagent was added for 4 hours of incubation, and absorbance at 570 nm was measured to calculate IC50 values[3]
- Apoptosis detection assay (Annexin V-FITC/PI double staining method): HeLa cells were seeded in 6-well plates, treated with 2~5 μM Nedaplatin for 48 hours, collected, stained with Annexin V-FITC and PI, and the proportion of apoptotic cells was detected by flow cytometry[5]
- Cell cycle analysis assay: A549 cells were treated with 3 μM Nedaplatin for 24 hours, fixed with ethanol, stained with PI, and the distribution of cells in each cycle was analyzed by flow cytometry[3]
- Western blot assay: SK-OV-3 cells were treated with 4 μM Nedaplatin for 24 hours, total protein was extracted and subjected to SDS-PAGE electrophoresis. After membrane transfer, incubation with caspase-3 and PARP primary antibodies was performed, followed by secondary antibody incubation and development to detect protein expression and cleavage[5]
- Colony formation assay: HCT116 cells were seeded in 6-well plates at 500 cells/well, treated with 1~5 μM Nedaplatin, stained with crystal violet after 14 days of culture, and the number of colonies was counted to calculate the inhibition rate[4]

Tumor-bearing athymic BALB/c nude mice with Ma44 or NCI-H460 cells
10 mg/kg or 20 mg/kg
Administered via i.v.

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Human lung cancer xenograft model (BALB/c nude mice): 2×10⁶ A549 cells were subcutaneously inoculated into the right back of mice, and drug administration started when the tumor volume reached 100 mm³. Nedaplatin was dissolved in normal saline to prepare a 0.8 mg/mL solution, administered intravenously at 4 mg/kg or 8 mg/kg once weekly for 3 consecutive weeks. Tumor volume was measured every 3 days, and tumors were excised and weighed at the end of the experiment[3]
- Human gastric cancer xenograft model (nude mice): 1×10⁷ MKN-45 cells were subcutaneously inoculated, and drug administration started when the tumor volume reached 150 mm³. The drug was dissolved in 5% glucose solution, administered intraperitoneally at 6 mg/kg once every two weeks for 2 consecutive times. Mouse body weight and tumor growth were monitored, and survival time was recorded[4]
- Cisplatin-resistant tumor model (A549/cis xenograft nude mice): 2×10⁶ A549/cis cells were subcutaneously inoculated, and when the tumor volume reached 120 mm³, Nedaplatin 8 mg/kg was administered intravenously once weekly for 3 consecutive weeks. A cisplatin control group (8 mg/kg) was set to compare the tumor inhibition effect[3]
- Combination therapy model (lung cancer xenograft nude mice): After A549 cell inoculation, Nedaplatin (4 mg/kg, intravenous injection, once weekly) was combined with paclitaxel (10 mg/kg, intraperitoneal injection, once weekly) for 3 consecutive weeks. Tumor volume was measured every 3 days to evaluate the synergistic effect[6]

Absorption, Distribution and Excretion
Most of the platinum in nedaplatin is excreted in the urine (59.6%). The volume of distribution of free platinum is 12.0 L. The clearance rate of free platinum is 4.47 L/h.

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Absorption: The oral bioavailability in rats is <5%, and the clinical administration is mainly by intravenous injection; after intravenous injection of 8 mg/kg, the peak plasma concentration (Cmax) is 28 μg/mL, and the time to peak concentration (Tmax) is 0.5 hours [1]
-Distribution: The drug concentration in tumor tissue is high. After intravenous injection of 8 mg/kg, the drug concentration in mouse tumor tissue can reach 3.1 times the plasma concentration; it is mainly distributed in the liver, kidney, spleen and tumor tissue, and the concentration in brain tissue is extremely low (<2% of plasma concentration) [1]
-Metabolism: It is mainly metabolized in the liver. Platinum ligands are removed by hydrolysis to form inactive platinum metabolites; the metabolic half-life is about 3.5 hours [1]
-Excretion: Within 72 hours after administration to rats, the amount excreted in urine accounts for 60%~65% of the administered dose, the amount excreted in feces accounts for 15%~20%, and the original drug accounts for 40% of the amount excreted in urine [1]
- Half-life: The elimination half-life (t1/2β) after intravenous injection in rats was 6.8 hours; after intravenous injection of 80 mg/m² in humans, t1/2β was 9.2 hours [1]
- Plasma protein binding rate: In vitro experiments showed that the plasma protein binding rate of this drug in human plasma was 75%~80%, mainly binding to albumin [1]

Protein Binding
Approximately 50% of the platinum in nedaplatin appears to bind to human plasma proteins.

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Nephrotoxicity: After intravenous injection of 16 mg/kg nedaplatin in rats, serum creatinine and blood urea nitrogen levels increased by 25%~30% compared with the control group, and were significantly lower than those in the cisplatin group (increased by 80%~90%); no tubular necrosis was observed after long-term administration (8 mg/kg, once a week for 4 weeks) [1]
- Bone marrow suppression: After intravenous injection of 10 mg/kg in mice, the white blood cell count decreased from 11×10⁹/L to 4.2×10⁹/L, and the platelet count decreased from 380×10⁹/L to 130×10⁹/L, and returned to normal 3 weeks after drug withdrawal [4]
- Gastrointestinal toxicity: After intravenous injection of 8 mg/kg in dogs, mild vomiting and diarrhea occurred, with an incidence of about 15%, and no serious gastrointestinal bleeding or ulceration occurred [1]
- Median lethal dose (LD50): The LD50 of intravenous injection in mice was 45 mg/kg, intraperitoneal LD50 is 58 mg/kg [1]
- Ototoxicity: After long-term administration (10 mg/kg, for 4 weeks), no significant increase in the auditory brainstem response threshold was observed in rats, and no cisplatin-like ototoxicity was observed [6]
- Drug interaction: In vitro experiments showed that the drug does not inhibit CYP450 enzymes (CYP3A4, CYP2D6, etc.), and no obvious pharmacokinetic interaction was observed when used in combination with paclitaxel or 5-fluorouracil [6]

[1]. Cancer Chemother Pharmacol . 1997;39(6):493-7.

[2]. Dalton Trans . 2010 Sep 21;39(35):8113-27.

[3]. Clin Cancer Res . 2001 Jan;7(1):202-9.

[4]. Eur J Cancer . 1998 Oct;34(11):1796-801.

[5]. Jpn J Cancer Res . 2001 Jan;92(1):51-8.

[6]. Cancer Res . 2014 Jul 15;74(14):3913-22.

Nedaplatin is a second-generation platinum analog. It has lower nephrotoxicity than [DB00515], but comparable efficacy. It was approved for marketing in Japan in 1995. Nedaplatin is a second-generation cisplatin analog with antitumor activity. It contains a novel cyclic structure in which glycolic acid binds to platinum atoms via bidentate ligands. Nedaplatin forms active platinum complexes that bind to nucleophilic groups in DNA, leading to intra- and inter-strand crosslinks and ultimately inducing apoptosis and cell death. Compared to cisplatin and carboplatin, this drug appears to have lower nephrotoxicity and neurotoxicity. Indications: For the treatment of non-small cell lung cancer, small cell lung cancer, esophageal cancer, and head and neck tumors. Mechanism of Action: As a platinum analog, nedaplatin's mechanism of action is likely similar to [DB00515], and the following mechanism description is based on [DB00515]. After entering the cell, nedaplatin is hydrolyzed to its active form and forms a complex with water molecules. This active form binds to nucleophiles in the cytoplasm, such as glutathione and other cysteine-rich proteins, causing cells to lose antioxidant proteins, thereby increasing oxidative stress. Nedaplatin can also bind to purine nucleotides in DNA. Its active form allows cross-linking at both binding sites. High-mobility group box 1 and 2 induce apoptosis upon stimulation by guanine cross-linking, and their binding can protect cross-linked DNA from repair mechanisms. Mismatch repair (MMR) protein complexes can also recognize DNA distortions caused by platinum-based complexes and attempt to repair the DNA. When MMR complexes attempt to remove platinum-based cross-links, single-strand breaks occur. After repair failure, MMR complexes induce apoptosis. Single-strand breaks in DNA are more likely to form lethal double-strand breaks during radiotherapy, resulting in the radiosensitizing effect of nedaplatin.

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Pharmacodynamics
Nedaplatin damages DNA and induces cancer cell death. It also has a radiosensitizing effect, increasing the sensitivity of affected cells to radiotherapy.

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Mechanism of Action: Nedaplatin is a second-generation platinum-based chemotherapy drug. After entering the cell, platinum drugs release platinum ions, which form intra- or inter-strand crosslinks with tumor cell DNA, destroying the structure and function of DNA. At the same time, they inhibit DNA topoisomerase II activity, block DNA repair and replication, and induce tumor cell apoptosis [2]
- Indications: Used to treat malignant tumors such as non-small cell lung cancer, esophageal cancer, gastric cancer and cervical cancer [4]
- Drug resistance mechanism: Some tumor cells develop drug resistance by upregulating the expression of DNA repair enzymes (such as ERCC1) and enhancing glutathione-mediated drug detoxification [6]
- Clinical advantages: Compared with cisplatin, nephrotoxicity and ototoxicity are significantly reduced, gastrointestinal reactions are milder, and patients tolerate better [1]
- Dosage: The recommended clinical dose for adults is 80~100 mg/m², intravenous infusion, for a course of treatment of 3~4 weeks [3]

C2H8N2O3PT

303.17

303.018

C, 7.98; H, 2.01; N, 9.30; O, 15.94; Pt, 64.78

95734-82-0

72120

Light yellow to yellow solid powder

265.6ºC at 760 mmHg

128.7ºC

0.12

4

5

0

8

50.5

0

[Pt+2].O([H])C([H])([H])C(=O)O[H].[N-]([H])[H].[N-]([H])[H]

ZAXCMPAWRCMABN-UHFFFAOYSA-N

InChI=1S/C2H4O3.2H3N.Pt/c3-1-2(4)5;;;/h3H,1H2,(H,4,5);2*1H3;

azane;2-hydroxyacetic acid;platinum

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: This product is not stable in solution, please use freshly prepared working solution for optimal results.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Solubility in Formulation 1: 7.14 mg/mL (23.55 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication (<60°C).

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 (Please use freshly prepared in vivo formulations for optimal results.)