LDH-A/lactate dehydrogenase-A

In NPCs, oxalic acid (10 μM; 24-72 h) suppresses cell growth in ways that are dose- and time-dependent [2]. In CNE-1 and CNE-2 cells, sodium oxalate (0–100 mM; 24 hours) promotes the G2/M phase of the cell life cycle [2]. By using caspase-3, sodium oxalate (0-100 mM; 48 hours) raises the levels of reactive oxygen species (ROS) in NPC cells [2].

When paired with radiation therapy, sodium oxalate (750 mg/kg; once daily; 3 weeks) tumor treatment can enhance the inhibitory impact in vivo [2].

Cell Proliferation Analysis[2]
Cell Types: CNE-1 and CNE activation and pathway-induced ROS levels in NPC cells[2]. -2 Cell
Tested Concentrations: 10 μM
Incubation Duration: 24-72 hrs (hours)
Experimental Results: In CNE-1 and CNE-2 cancer cells, the IC50 at 24, 48 and 72 hrs (hours) were 74.6, 32.4 and 17.8 mM and 62.3, 44.5, respectively. 31.6 mM, respectively.

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Apoptosis analysis [2]
Cell Types: Nasopharyngeal carcinoma cells
Tested Concentrations: 0, 20, 50, 100 mM
Incubation Duration: 48 hrs (hours)
Experimental Results: There was a dose-dependent increase in early and late apoptotic cells. Increases the expression of pro-apoptotic Bax and cleaved-caspase-3, while reducing the anti-apoptotic signals of Bcl-2 and pro-caspase-3.

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Cell cycle analysis [2]
Cell Types: CNE-1 and CNE-2 Cell
Tested Concentrations: 0, 20, 50 and 100 mM
Incubation Duration: 24 hrs (hours)
Experimental Results: Dose-dependent increase in CNE-1 and CNE- numbers 2 cells in G2/M stage.

Animal/Disease Models: Female Balb/c nude mice were injected with CNE-1 cells [2].
Doses: 750 mg/kg
Route of Administration: intraperitoneal (ip) injection; 750 mg/kg; one time/day; 3 weeks
Experimental Results: Inhibition of tumor growth compared with oxalate alone or irradiation alone.

[1]. LDH-A inhibitors as remedies to enhance the anticancer effects of PARP inhibitors in ovarian cancer cells. Aging (Albany NY). 2021 Dec 16;13(24):25920-25930.

[2]. Inhibition of LDH-A by oxamate induces G2/M arrest, apoptosis and increases radiosensitivity in nasopharyngeal carcinoma cells. Oncol Rep. 2013 Dec;30(6):2983-91.

[3]. Targeting lactate dehydrogenase‑A promotes docetaxel‑induced cytotoxicity predominantly in castration‑resistant prostate cancer cells. Oncol Rep. 2019 Jul;42(1):224-230.

Ovarian cancer is one of the deadliest gynecological malignancies. Studies have shown that PARP inhibitors can selectively target BRCA-mutated ovarian cancer and also have some inhibitory effect on ovarian cancer without BRCA mutations. However, their mechanism of action remains unclear. This study used wild-type BRCA ovarian cancer cell lines (A2780 and SKOV3). The results showed that PARP inhibitors alone (olaparib or AG14361) significantly inhibited the proliferation of A2780 cells, but had little inhibitory effect on the proliferation of SKOV3 cells. We analyzed differentially expressed genes using RNA sequencing technology and found that PARP inhibitors increased LDH-A expression in SKOV3 cells, and verified this result by RT-PCR. We further investigated whether LDH-A inhibition could enhance the inhibitory effect of PARP inhibitors on ovarian cancer without BRCA mutations using oxalate (an LDH-A specific inhibitor). Cell proliferation, migration, and invasion were detected using the CCK-8 assay, scratch assay, and Transwell assay, respectively. Olaparib and AG14361 significantly inhibited the proliferation/invasion of A2780 cells, but had no significant effect on SKOV3 cells. LDH-A inhibitors significantly enhanced the inhibitory effect of PARP inhibitors on A2780 and SKOV3 cells. Therefore, high LDH-A expression levels affect the inhibitory effect of PARP inhibitors on ovarian cancer carrying wild-type BRCA, while LDH-A inhibition significantly enhances this inhibitory effect. [1] The increased glucose consumption rate and dependence on aerobic glycolysis to produce ATP in cancer cells, known as the Wahlberg effect, has been observed for some time. Changes in energy metabolism in cancer cells provide an attractive opportunity to develop novel cancer treatment strategies. Lactate dehydrogenase (LDH) catalyzes the conversion of pyruvate to lactate and plays a crucial role in glycolysis. It has been reported that LDH-A expression levels are elevated in the serum of patients with head and neck cancer cells and nasopharyngeal carcinoma (NPC) and are associated with poor prognosis. However, the effect of LDH-A inhibitors on NPC cells is still unclear. This study found that oxalamide, a classic inhibitor of LDH-A, inhibited the proliferation of two NPC cancer cell lines, CNE-1 and CNE-2, in a dose- and time-dependent manner. Oxalide induced cell cycle arrest in the G2/M phase by downregulating the CDK1/cyclin B1 pathway and promoted apoptosis by enhancing the generation of mitochondrial reactive oxygen species (ROS). N-acetylcysteine is a specific reactive oxygen species scavenger that significantly blocked the growth inhibition induced by oxalamide. We also found that oxalamide increased the sensitivity of the two nasopharyngeal carcinoma cell lines to ionizing radiation. In addition, we validated similar results in a tumor xenograft model. In summary, these results suggest that LDH-A may be a promising target for the treatment of nasopharyngeal carcinoma. [2]

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Docetaxel (DOC) is one of the most effective chemotherapy drugs for the treatment of castration-resistant prostate cancer (CRPC). Despite significant initial clinical efficacy, most patients eventually develop resistance to DOC. In tumor metabolism, tumors preferentially utilize anaerobic metabolism, and lactate dehydrogenase (LDH) plays a crucial role. LDH controls the conversion of pyruvate to lactate, and LDH-A is one of the major isoenzymes of LDH, responsible for regulating this metabolic process. This study investigated the role of LDH-A in drug resistance in human prostate cancer (PC) by analyzing four prostate cancer cell lines (including castration-induced lines PC3, DU145, LNCaP, and LN-CSS, the latter being a hormone-refractory cell line established from LNCaP). Sodium oxalamide (SO) was used as a specific LDH-A inhibitor. Changes in LDH-A expression levels were analyzed using Western blotting. Cell growth and survival were assessed using the WST-1 assay. Cell cycle progression and apoptosis induction were assessed using flow cytometry combined with propidium iodide and Annexin V staining. LDH expression was closely related to the sensitivity of PC cells to docetaxel (DOC). SO inhibited PC cell growth, which was thought to be due to the inhibition of LDH-A expression. Synergistic cytotoxicity of DOC and SO combination therapy was observed in LN-CSS cells, but not in LNCaP cells. The combination therapy produced additive cytotoxicity in PC-3 and DU145 cells, leading to cell cycle arrest in the G2-M phase of LN-CSS cells and increasing the number of cells in the sub-G1 phase. SO promoted DOC-induced apoptosis in LN-CSS cells, partly due to its inhibition of DOC-induced increase in LDH-A expression. The results strongly suggest that LDH-A plays an important role in DOC resistance in late-stage PC cells, and that inhibiting LDH-A expression can increase the sensitivity of cells to DOC, especially in CRPC cells. This study may provide valuable information for the future development of targeted therapies for CRPC. [3]

C2H2NNAO3

111.0320

110.993

C, 21.64; H, 1.82; N, 12.62; Na, 20.71; O, 43.23

565-73-1

5242

Typically exists as White to off-white solids at room temperature

306.3ºC at 760 mmHg

300 °C

139ºC

1

3

1

7

90.9

0

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

RQVZIJIQDCGIKI-UHFFFAOYSA-M

InChI=1S/C2H3NO3.Na/c3-1(4)2(5)6;/h(H2,3,4)(H,5,6);/q;+1/p-1

sodium;oxamate

Oxamic acid sodium salt; Sodium 2-amino-2-oxoacetate; Oxamic acid, sodium salt; Aminooxoacetic acid sodium salt; Acetic acid, aminooxo-, monosodium salt; Oxamate (sodium);

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: Please store this product in a sealed and protected environment, avoid exposure to moisture.

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

H2O : ~12.5 mg/mL (~112.58 mM)
DMSO : ~3.23 mg/mL (~29.09 mM)

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.)