LDHA/lactate dehydrogenase A (Ki =8 μM)

Acetone-CoA pyruvylase, which is the substrate of FX-11 (9 μM) [2], is phosphorylated to demonstrate the activation of AMP. In P493 cells, FX-11 suppresses glycolysis and modifies cellular energy supplementing. In BxPc-3 and MIA PaCa-2 cells, FX-11 (0-100 μM, 72 h) limits cell growth [3].

FX-11 (42 μg/mouse; IP, once daily for 10–14 days) suppresses the formation of P493 tumors [2]. FX-11 (0–2 mg/kg, IP, once daily for three weeks) Standing

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In this study, researchers investigated whether the PKM2 activator, TEPP-46, and the LDHA inhibitor, FX-11, can be combined to inhibit in vitro and in vivo tumor growth in preclinical models of pancreatic cancer. They assessed PKM2 and LDHA expression, enzyme activity, and cell proliferation rate after treatment with TEPP-46, FX-11, or a combination of both. Efficacy was validated in vivo by evaluating tumor growth, PK and LDHA activity in plasma and tumors, and PKM2, LDHA, and Ki-67 expression in tumor tissues following treatment. Dual therapy synergistically inhibited pancreatic cancer cell proliferation and significantly delayed tumor growth in vivo without apparent toxicity. Treatment with TEPP-46 and FX-11 resulted in increased PK and reduced LDHA enzyme activity in plasma and tumor tissues and decreased PKM2 and LDHA expression in tumors, which was reflected by a decrease in tumor volume and proliferation. The targeting of glycolytic enzymes such as PKM2 and LDHA represents a promising therapeutic approach for the treatment of pancreatic cancer.[2]

Western Blot Analysis [2]
Cell Types: P493 Cell
Tested Concentrations: 9 μM
Incubation Duration: 24 hrs (hours), 48 hrs (hours)
Experimental Results: ATP levels diminished, accompanied by activation of AMP kinase and phosphorylation of its substrate acetyl-CoA carboxylase.

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Cell proliferation assay [3]
Cell Types: BxPc-3 and MIA PaCa-2 Cell
Tested Concentrations: 0-100 µM
Incubation Duration: 72 hrs (hours)
Experimental Results: diminished cell metabolic activity in a concentration-dependent manner, showing significant reduction in cell proliferation, BxPc- The IC50 values for 3 and MIA PaCa-2 cells were 49.27 µM and 60.54 µM, respectively.

Animal/Disease Models: Male SCID (severe combined immunodeficient) mouse and RH-Foxn1nu (nude) mice (human P493 B cell xenografts) [2]
Doses: 42 μg /mouse (2.1 mg/kg)
Route of Administration: IP; delays tumor growth [3]. one time/day for 10-14 days.
Experimental Results: Significant inhibition of tumor growth and inhibition of tumor xenograft progression.

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Animal/Disease Models: Immunocompromised CD-1 mice (6-8 weeks; 20-25 g, n=5 per group) [3]
Doses: 2 mg/kg, 1 mg/kg+15 mg/kg TEPP- 46. 2 mg/kg+30 mg/kg TEPP-46
Route of Administration: intraperitoneal (ip) injection (100 µL), daily, for 3 weeks
Experimental Results: LDHA activity in plasma and tumor lysates was Dramatically diminished; proliferation markers were Dramatically diminished The expression of Ki-67; a significant decrease in proliferation index was observed in tumor sections; and a significant delay in tumor growth.

[1]. EC Calvaresi. Small molecule inhibitors of lactate dehydrogenase a as an anticancer strategy. 2014.

[2]. Inhibition of lactate dehydrogenase A induces oxidative stress and inhibits tumor progression. Proc Natl Acad Sci U S A. 2010 Feb 2;107(5):2037-42.

[3]. Targeting Pyruvate Kinase M2 and Lactate Dehydrogenase A Is an Effective Combination Strategy for the Treatment of Pancreatic Cancer. Cancers (Basel). 2019 Sep 16;11(9):1372.

Due to genetic alterations and tumor hypoxia, many cancer cells take up large amounts of glucose and generate lactate via lactate dehydrogenase A (LDHA). LDHA is encoded by target genes of c-Myc and hypoxia-inducible factor (HIF-1). Previous studies have shown that decreased LDHA expression is associated with tumor initiation, but its role in tumor maintenance and progression remains unclear. Furthermore, the mechanism by which decreasing LDHA expression through interference or antisense RNA inhibits tumorigenesis is unknown. This study found that decreasing LDHA expression via siRNA or inhibiting LDHA activity using the small molecule inhibitor FX11 [3-dihydroxy-6-methyl-7-(phenylmethyl)-4-propylnaphthalene-1-carboxylic acid] both reduced ATP levels and induced significant oxidative stress and cell death, which were partially reversed by the antioxidant N-acetylcysteine. In addition, we demonstrated that FX11 can inhibit the progression of large human lymphomas and pancreatic cancer xenografts. When used in combination with the NAD(+) synthesis inhibitor FK866, FX11 induced lymphoma regression. Therefore, using FX11 to inhibit LDHA is a feasible and well-tolerated approach to treat LDHA-dependent tumors. Our study documents a treatment approach targeting the Wahlberg effect and demonstrates that oxidative stress and cancer metabolic phenotypes are key aspects of cancer biology that need to be considered for targeted cancer energy metabolism therapy. [2]

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In recent years, the use of cancer cell metabolism as an anticancer therapy strategy has attracted widespread attention. As early as the 1920s, German scientist Otto Wahlberg observed the vigorous consumption of glucose and high levels of aerobic glycolysis in cancer tissue, a phenomenon now known as the Wahlberg effect. Today, we recognize that the Wahlberg effect is mediated by a variety of complex factors, including the overexpression of the insulin-independent glucose transporter GLUT-1 and the overexpression of various glycolytic enzymes, such as lactate dehydrogenase A (LDH-A). As the terminal enzyme of glycolysis, LDH-A catalyzes the reversible conversion of pyruvate to lactate and oxidizes NADH to NAD+ in the process. Most of the lactate produced by this reaction is secreted into the tumor microenvironment, acidifying the surrounding tissue and helping the tumor evade the attack of immune cells. The oxidation of NADH to NAD+ can replenish NAD+ in the absence or impaired function of oxidative metabolism, thereby maintaining the continuous ATP production from glycolysis. Cell culture and in vivo studies have shown that LDH-A knockdown (using RNA interference) significantly reduces cell and tumor proliferation, suggesting that LDH-A may be a viable anticancer target. Although various in vitro LDH-A inhibitors exist, there is still a need for a highly efficient and selective small molecule inhibitor that can act both intracellularly and in vivo. This article reports the development and biological evaluation of N-hydroxyindole LDH-A inhibitors, including a series of novel dual Warburg-targeted glucose-coupled LDH-A inhibitors developed in collaboration between the Hergenrother and Minutolo laboratories. This article also discusses the development of novel assays for evaluating the relative cellular uptake, cellular lactate production, and competition for cell entry with 13C glucose of the NHI series compounds. Furthermore, this article reports a direct cellular activity comparison of the most promising NHI series compounds with reported in vitro LDH-A inhibitors. Finally, this paper also discusses the study of the interaction between compounds and LDH-A in cell lysates and intact cells using CETSA and DARTS techniques [1].

C22H22O4MO

350.4077

350.152

C, 75.41; H, 6.33; O, 18.26

213971-34-7

10498042

White to off-white solid powder

4.8

3

4

5

26

473

0

LVPYVYFMCKYFCZ-UHFFFAOYSA-N

InChI=1S/C22H22O4/c1-3-7-16-17-10-13(2)15(11-14-8-5-4-6-9-14)12-18(17)19(22(25)26)21(24)20(16)23/h4-6,8-10,12,23-24H,3,7,11H2,1-2H3,(H,25,26)

7-Benzyl-2,3-dihydroxy-6-methyl-4-propylnaphthalene-1-carboxylic acid

FX 11; FX11; LDHA Inhibitor FX11; FX11; 7-benzyl-2,3-dihydroxy-6-methyl-4-propyl-naphthalene-1-carboxylic Acid; 2,3-Dihydroxy-6-methyl-7-(phenylmethyl)-4-propyl-1-naphthalenecarboxylic Acid; CHEMBL126519; 7-benzyl-2,3-dihydroxy-6-methyl-4-propylnaphthalene-1-carboxylic acid; FX-11

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)

DMSO : ~250 mg/mL (~713.45 mM)

Solubility in Formulation 1: ≥ 2.08 mg/mL (5.94 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 2.08 mg/mL (5.94 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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