DNA alkylating/crosslinking

(rel)-Oxaliplatin (2-128 μM; 24–72 hours; HCC, HCCLM3, and Hep3B cells) causes apoptosis and inhibits cell growth [1]. DNA cross-linking (ISC) and DNA-protein cross-linking (DPC) are two examples of the primary and secondary DNA damage that (rel)-Oxaliplatin (10 μM; 15-240 min; CEM cells) induces[2]. (rel)-Oxaliplatin (0.01 to 100 μM; 24 hours) efficiently inhibits the following cell lines: melanoma cell lines SK-MEL-2 and HT-144; ovarian cancer cell line A2780; colon cancer cell line HT-29; glioblastoma cell lines U-373MG and U-87MG; bladder cancer cell lines RT4 and TCCSUP; IC50 are 11 μM, 15 μM, 0.17 μM, 0.97 μM, 2.95 μM, 17.6 μM, 30.9 μM, and 7.85 μM, respectively [3]Cells

(rel)-Oxaliplatin (5–10 mg/kg; intraperitoneally; for 32 days; in nude mice) suppresses the formation of tumors [1].

Cell viability assay [1] Cells
Cell Types: HCC, HCCLM3 and Hep3B Cell
Tested Concentrations: 24, 48 and 72 hrs (hours)
Incubation Duration: 2, 4, 8, 16, 32, 64 and 128 μM
Experimental Results: Cell viability diminished in a dose- and time-dependent manner .

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Western Blot Analysis [1]
Cell Types: HCCLM3 and Hep3B cells
Tested Concentrations: 48 hrs (hours)
Incubation Duration: 10 μM
Experimental Results: The expression of Bcl-2 and Bcl-xL was down-regulated, and the expression of Bax was up-regulated.

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Cell cycle analysis [1]
Cell Types: HCCLM3 and Hep3B cells
Tested Concentrations: 24 hrs (hours)
Incubation Duration: 10 μM
Experimental Results: Increased percentage of apoptotic cells (17.70% for HCCLM3 cells; 21.19% for Hep3B cells).

Animal/Disease Models: Nude mice [1]
Doses: 5 and 10 mg/kg
Route of Administration: intraperitoneal (ip) injection; continued for 32 days
Experimental Results: Tumor volume reduction in HCCLM3 tumor xenografts.

[1]. Woynarowski JM, et, al. Oxaliplatin-induced damage of cellular DNA. Mol Pharmacol. 2000 Nov;58(5):920-7.
[2]. Wang Z, et, al. Oxaliplatin induces apoptosis in hepatocellular carcinoma cells and inhibits tumor growth. Expert Opin Investig Drugs. 2009 Nov;18(11):1595-604.
[3]. Pendyala L, et, al. In vitro cytotoxicity, protein binding, red blood cell partitioning, and biotransformation of oxaliplatin. Cancer Res. 1993 Dec 15;53(24):5970-6.

Objective: Oxaliplatin, a platinum-based chemotherapy drug, possesses broad-spectrum antitumor activity. Currently, detailed data on the effects of oxaliplatin on hepatocellular carcinoma (HCC) cells are lacking. This study aimed to investigate the inhibitory effects of oxaliplatin on the proliferation of HCCLM3 and Hep3B cells in vitro and in vivo. Methods: Cell viability was assessed using the MTT assay, and apoptosis was detected by flow cytometry and transmission electron microscopy. Gene chip analysis, quantitative reverse transcription-PCR, and Western blotting were used to detect the expression of apoptosis-related proteins in HCCLM3 cells. Furthermore, the in vivo effects of oxaliplatin were investigated using a xenograft tumor model. Results: Oxaliplatin inhibited the growth of HCCLM3 and Hep3B cells. Using flow cytometry, transmission electron microscopy, and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL), we found that apoptosis is the main mechanism by which oxaliplatin inhibits tumor progression. Microarray analysis, quantitative reverse transcription PCR and Western blotting further confirmed that during oxaliplatin-induced apoptosis, the expression of anti-apoptotic proteins Bcl-2 and Bcl-xL was downregulated, while the expression of pro-apoptotic protein Bax was upregulated. Conclusion: The anti-proliferative effect of oxaliplatin on liver cancer cells is due to its induction of apoptosis. Therefore, oxaliplatin may be an effective drug for the treatment of liver cancer, and its application deserves further in-depth research. [1] Objective: Platinum-based chemotherapy drug oxaliplatin has broad anti-tumor activity. To date, there is no detailed data on the effect of oxaliplatin on hepatocellular carcinoma (HCC) cells. This article studies the anti-proliferative effect of oxaliplatin on HCCLM3 and Hep3B cells in vitro and in vivo. Methods: Cell viability was assessed by MTT assay, and apoptosis was detected by flow cytometry and transmission electron microscopy. The expression of apoptosis-related proteins in HCCLM3 cells was detected by microarray analysis, quantitative reverse transcription-PCR and Western blotting. In addition, the effects of oxaliplatin in vivo were studied using a xenograft tumor model. Results: Oxaliplatin inhibited the growth of HCCLM3 and Hep3B cells. By flow cytometry, transmission electron microscopy and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay, we found that apoptosis was the main mechanism by which oxaliplatin inhibited tumor progression. Microarray analysis, quantitative reverse transcription-PCR and Western blot analysis further confirmed that during oxaliplatin-induced apoptosis, the expression of anti-apoptotic proteins Bcl-2 and Bcl-xL was downregulated, while the expression of pro-apoptotic protein Bax was upregulated. Conclusion: The anti-proliferative effect of oxaliplatin on liver cancer cells is due to its induction of apoptosis. Therefore, oxaliplatin may be an effective drug for the treatment of liver cancer, and its application deserves further in-depth research. [2] This study investigated the in vitro cytotoxicity, protein binding, platinum distribution from whole blood to erythrocytes, platinum exchange from erythrocytes to plasma, and in vitro biotransformation in plasma of the novel non-nephrotoxic platinum analog oxaliplatin. Cytotoxicity studies were conducted on a range of human tumor cell lines derived from ovarian cancer (A2780, A2780/cp), bladder cancer (TCCSUP, RT4), colon cancer (HT-29), melanoma (SKMEL-2, HTB144), and glioma (U373MG and U87MG). The relative potency of the five platinum-based complexes was: oxaliplatin = tetraplatin > cisplatin > isopropylplatin > carboplatin. Oxaliplatin was effective against HT-29 cells and showed very low cross-resistance with cisplatin against A2780/cp cells. Two bladder cancer cell lines, two melanoma cell lines, and one of two glioblastoma cell lines were resistant to both oxaliplatin and tetraplatin. The cytotoxicity profiles of the oxaliplatin-tetraplatin and cisplatin-carboplatin groups showed statistically significant correlations according to Spearman's rank correlation test. Oxaliplatin's protein-binding capacity is similar to that of cisplatin and tetraplatin; 85-88% of the platinum in oxaliplatin (5, 10, or 20 μg/mL) binds to plasma proteins within 5 hours after administration, with a mean half-life of 1.71 ± 0.06 hours. When oxaliplatin is incubated in whole blood (5, 10, and 20 μg/mL), erythrocytes absorb 37.1 ± 2.1% of the total platinum (maximum absorption) within 2 hours, and this platinum cannot be exchanged into the plasma. Therefore, platinum bound to erythrocytes does not serve as a drug reservoir. In plasma, oxaliplatin remains unchanged at 0.5 hours, but at 1 hour, 30% of the total platinum in plasma appears in a peak with a retention time similar to that of dichloroplatin(II), the major biotransformation product of tetraplatin (trans-1,2-diaminocyclohexane). At 2 hours, (trans-1,2-diaminocyclohexane)dichloroplatin(II) and three other platinum-containing peaks were detected, but oxaliplatin, which remained unchanged, was not detected. All platinum eluted in a single peak near the solvent front at 4 hours. The striking similarity in cytotoxicity between oxaliplatin and tetraplatin may be due to the formation of (trans-1,2-diaminocyclohexane)dichloroplatin(II) in tissue culture medium. [3]

C8H12N2O4PT

395.269885063171

397.06

C, 24.19; H, 3.55; N, 7.05; O, 16.11; Pt, 49.10

63121-00-6

Oxaliplatin;61825-94-3;Oxaliplatin-d10;1132819-16-9

Typically exists as solid at room temperature

193.6ºC at 760 mmHg

75ºC

0.614

4

6

0

15

191

2

O=C1C(=O)O[Pt]2(NC3CCCCC3N2)O1

DRMCATBEKSVAPL-BNTLRKBRSA-N

InChI=1S/C6H12N2.C2H2O4.Pt/c7-5-3-1-2-4-6(5)8;3-1(4)2(5)6;/h5-8H,1-4H2;(H,3,4)(H,5,6);/q-2;;+2/t5-,6-;;/m1../s1

trans-1,2-Cyclohexanediamine, platinum complex; Platinum, [rel-(1R,2R)-1,2-cyclohexanediamine-kappaN1,kappaN2][ethanedioato(2-)-kappaO1,kappaO2]-, (SP-4-2)-;

trans-1,2-Cyclohexanediamine, platinum complex; Platinum, [rel-(1R,2R)-1,2-cyclohexanediamine-kappaN1,kappaN2][ethanedioato(2-)-kappaO1,kappaO2]-, (SP-4-2)-; 63121-00-6

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

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

Note: Do not dissolve Oxaliplatin in DMSO, as platinum-based drugs are prone to deactivation in DMSO. Additionally, Oxaliplatin is not stable in solution and should be prepared immediately before use. DMSO has been reported to significantly inhibit or completely abolish the biological activity of Oxaliplatin.

Note: Oxaliplatin is generally not recommended to be dissolved in DMSO, as platinum-based drugs are prone to deactivation in DMSO. Additionally, Oxaliplatin is not stable in solution and should be prepared immediately before use.

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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