FX11

LDHA/lactate dehydrogenase A (Ki =8 μM)

丙酮辅酶 A 丙酮酰酶是 FX-11 (9 μM) [2] 的底物，它被磷酸化以证明 AMP 的激活。在 P493 细胞中，FX-11 抑制糖酵解并改变细胞能量补充。在 BxPc-3 和 MIA PaCa-2 细胞中，FX-11（0-100 μM，72 小时）限制细胞生长 [3]。

FX-11（42 μg/小鼠；IP，每天一次，持续 10-14 天）抑制 P493 肿瘤的形成 [2]。 FX-11（0–2 mg/kg，IP，每日一次，持续三周）

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在这项研究中，研究人员研究了PKM2激活剂TEPP-46和LDHA抑制剂FX-11是否可以联合抑制胰腺癌临床前模型的体外和体内肿瘤生长。他们评估了用TEPP-46、FX-11或两者联合处理后的PKM2和LDHA表达、酶活性和细胞增殖率。在体内通过评估肿瘤生长情况、血浆和肿瘤中PK和LDHA活性以及治疗后肿瘤组织中PKM2、LDHA和Ki-67的表达来验证疗效。双重治疗在体内协同抑制胰腺癌细胞增殖，显著延缓肿瘤生长，无明显毒性。TEPP-46和FX-11处理后，血浆和肿瘤组织中PK和LDHA酶活性增加，LDHA酶活性降低，肿瘤中PKM2和LDHA表达降低，表现为肿瘤体积和增殖减少。靶向糖酵解酶如PKM2和LDHA是治疗胰腺癌的一种很有前景的治疗方法。[2]

蛋白质印迹分析 [2]
细胞类型： P493 细胞
测试浓度： 9 μM
孵育时间： 24 hrs（小时）、48 hrs（小时）
实验结果： ATP 水平降低，伴随着 AMP 激酶的激活及其底物乙酰辅酶 A 羧化酶的磷酸化。

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细胞增殖测定 [3]
细胞类型： BxPc-3 和 MIA PaCa-2 细胞
测试浓度： 0-100 µM
孵育持续时间：72 hrs（小时）
实验结果：以浓度依赖性方式降低细胞代谢活性，显示显着降低细胞增殖，BxPc-3 和 MIA PaCa-2 细胞的 IC50 值分别为 49.27 µM 和 60.54 µM。

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.

As the result of genetic alterations and tumor hypoxia, many cancer cells avidly take up glucose and generate lactate through lactate dehydrogenase A (LDHA), which is encoded by a target gene of c-Myc and hypoxia-inducible factor (HIF-1). Previous studies with reduction of LDHA expression indicate that LDHA is involved in tumor initiation, but its role in tumor maintenance and progression has not been established. Furthermore, how reduction of LDHA expression by interference or antisense RNA inhibits tumorigenesis is not well understood. Here, we report that reduction of LDHA by siRNA or its inhibition by a small-molecule inhibitor (FX11 [3-dihydroxy-6-methyl-7-(phenylmethyl)-4-propylnaphthalene-1-carboxylic acid]) reduced ATP levels and induced significant oxidative stress and cell death that could be partially reversed by the antioxidant N-acetylcysteine. Furthermore, we document that FX11 inhibited the progression of sizable human lymphoma and pancreatic cancer xenografts. When used in combination with the NAD(+) synthesis inhibitor FK866, FX11 induced lymphoma regression. Hence, inhibition of LDHA with FX11 is an achievable and tolerable treatment for LDHA-dependent tumors. Our studies document a therapeutical approach to the Warburg effect and demonstrate that oxidative stress and metabolic phenotyping of cancers are critical aspects of cancer biology to consider for the therapeutical targeting of cancer energy metabolism.[2]

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Exploiting cancer cell metabolism as an anticancer therapeutic strategy has garnered much attention in recent years. As early as the 1920s, German scientist Otto Warburg observed cancer tissues’ avid glucose consumption and high rates of aerobic glycolysis, a phenomenon now known as the Warburg effect. Today, we understand the Warburg effect is mediated by a number of complex factors, including overexpression of the insulin-independent glucose transporter GLUT-1 and overexpression of various glycolytic enzymes, including lactate dehydrogenase A (LDH-A). As the terminal enzyme of glycolysis, LDH-A catalyzes the reversible conversion of pyruvate to lactate, and in doing so, oxidizes NADH to NAD+ . The lactate produced by this reaction is largely excreted into the tumor microenvironment, where it acidifies surrounding tissues and helps the tumor evade destruction by immune cells. The oxidation of NADH to NAD+ allows for continued ATP production through glycolysis by replenishing NAD+ in the absence, or reduced function, of oxidative metabolism. Cell culture and in vivo studies of LDH-A knockdown (using RNA interference) have been shown to lead to substantial decreases in cell and tumor proliferation, thus providing evidence that LDH-A would be a viable anticancer target. While various in vitro LDH-A inhibitors exist, there is a need for a potent and selective small molecule inhibitor that functions both in cells and in vivo. Here, the development and biological assessment of the N-hydroxyindole class of LDH-A inhibitors, including a series of novel dual-Warburg targeting glucose-conjugated LDH-A inhibitors, developed through a collaboration between the Hergenrother and Minutolo laboratories, is reported. The development of novel assays to assess the relative cell uptake, cell lactate production, and competition with 13C glucose for cellular entry, of NHI series compounds are also discussed. Head-to-head cellular assessments of the most promising NHI series compounds alongside literature-reported in vitro inhibitors of LDH-A are reported. Finally, efforts to directly probe the interactions of compounds with LDH-A in cell lysate and whole cells using CETSA and DARTS techniques are discussed[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

77.76

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)

配方 1 中的溶解度: ≥ 2.08 mg/mL (5.94 mM) (饱和度未知) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将100 μL 20.8 mg/mL澄清DMSO储备液加入400 μL PEG300中，混匀；然后向上述溶液中加入50 μL Tween-80，混匀；加入450 μL生理盐水定容至1 mL。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

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配方 2 中的溶解度: ≥ 2.08 mg/mL (5.94 mM) (饱和度未知) in 10% DMSO + 90% Corn Oil (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 20.8 mg/mL 澄清 DMSO 储备液加入到 900 μL 玉米油中并混合均匀。

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
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6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。