Betulinic acid (ALS357; Lupatic acid)   中文名称: 白桦脂酸

Topoisomerase I ( IC50 = 5 μM ); HIV-1 ( IC50 = 1.4 μM ); NF-κB

体外活性：桦木酸具有抗 HIV 活性，可阻断 H9 淋巴细胞中的 HIV 复制，EC50 值为 1.4 μM，并抑制未感染的 H9 细胞生长，IC50 值为 13 μM。桦木酸还通过抑制氨肽酶 N 活性发挥抗黑色素瘤作用，IC50 为 7.3 μM。此外，桦木酸可抑制真核拓扑异构酶 I 活性，IC50 为 5 μM，可作为抗肿瘤药物的有力候选药物。激酶测定：在结合缓冲液中存在或不存在桦木酸的情况下，将寡核苷酸 1 和寡核苷酸 2 的退火 25 聚体双链体与 25 或 50 单位的大鼠肝拓扑异构酶 I 在 8 °C 下孵育 15 分钟。孵育后，将反应混合物在 7% 非变性聚丙烯酰胺凝胶中于 4 °C 的 0.167 × TBE 缓冲液中进行电泳，并用溴化乙锭对 DNA 条带进行染色。细胞测定：测定中使用CCK-8。 MDA-MB-231 细胞以 2 × 103 个细胞/孔的密度在 96 孔板中培养，然后用 DMSO 载体或 100 µL 培养基中的各种浓度（5 µM 至 160 µM）的桦木酸进行处理，以达到指定的效果。次。处理期结束后，将每孔中的培养基更换为110μL含有10μL CCK-8混合物的培养基，并将板在37°C下孵育1小时30分钟。使用酶标仪测量 450 nm 波长处的吸光度。

桦木酸可抑制携带 LNCaP 细胞的无胸腺裸鼠中前列腺癌细胞和肿瘤（异种移植物）的生长，部分原因是蛋白酶体依赖性 Sp1、Sp3、Sp4 和几个 Sp 调节基因的下调。

将由寡核苷酸 1 和寡核苷酸 2 组成的 25 聚体退火双链体与 25 或 50 单位的大鼠肝拓扑异构酶 I 一起进行 8°C 孵育，无论结合缓冲液中存在或不存在桦木酸。孵育后，反应混合物在 4 °C 下在 0.167 × TBE 缓冲液中的 7% 非变性聚丙烯酰胺凝胶中进行电泳。然后使用溴化乙锭对 DNA 带进行染色。

该测定使用 CCK-8。在 96 孔板中以 2 × 10³ 细胞/孔的密度生长后，用 DMSO 载体或不同浓度的桦木酸（5 µM 至 160 µM）处理 MDA-MB-231 细胞。 ）在 100 µL 介质中持续指定的持续时间。处理阶段后，将含有 10 µL CCK-8 混合物的 110 µL 培养基添加到每个孔中，并将板在 37°C 下孵育 1 小时 30 分钟。借助酶标仪，测定 450 nm 波长处的吸光度[2]。

[1]. Betulinic acid, a potent inhibitor of eukaryotic topoisomerase I: identification of the inhibitory step, the major functional group responsible and development of more potent derivatives. Med Sci Monit. 2002 Jul;8(7):BR254-65.

[2]. Betulinic acid induces apoptosis and ultrastructural changes in MDA-MB-231 breast cancer cells. Ultrastruct Pathol. 2018 Jan-Feb;42(1):49-54.

[3]. Betulinic acid alleviates dextran sulfate sodium-induced colitis and visceral pain in mice. Naunyn Schmiedebergs Arch Pharmacol. 2017 Dec 26.

[4]. Anti-AIDS agents--XXVII. Synthesis and anti-HIV activity of betulinic acid and dihydrobetulinic acid derivatives. Bioorg Med Chem. 1997 Dec;5(12):2133-43.

[5]. Betulinic acid as new activator of NF-kappaB: molecular mechanisms and implications for cancer therapy. Oncogene. 2005 Oct 20;24(46):6945-56.

Betulinic acid is a pentacyclic triterpenoid that is lupane having a double bond at position 20(29) as well as 3beta-hydroxy and 28-carboxy substituents. It is found in the bark and other plant parts of several species of plants including Syzygium claviflorum. It exhibits anti-HIV, antimalarial, antineoplastic and anti-inflammatory properties. It has a role as an EC 5.99.1.3 [DNA topoisomerase (ATP-hydrolysing)] inhibitor, an anti-HIV agent, an antimalarial, an anti-inflammatory agent, an antineoplastic agent and a plant metabolite. It is a pentacyclic triterpenoid and a hydroxy monocarboxylic acid. It derives from a hydride of a lupane.
Betulinic Acid has been used in trials studying the treatment of Dysplastic Nevus Syndrome.
Betulinic acid has been reported in Paeonia emodi, Bowdichia virgilioides, and other organisms with data available.
Betulinic Acid is a pentacyclic lupane-type triterpene derivative of betulin (isolated from the bark of Betula alba, the common white birch) with antiinflammatory, anti-HIV and antineoplastic activities. Betulinic acid induces apoptosis through induction of changes in mitochondrial membrane potential, production of reactive oxygen species, and opening of mitochondrial permeability transition pores, resulting in the release of mitochondrial apogenic factors, activation of caspases, and DNA fragmentation. Although originally thought to exhibit specific cytotoxicity against melanoma cells, this agent has been found to be cytotoxic against non-melanoma tumor cell types including neuroectodermal and brain tumor cells.
A lupane-type triterpene derivative of betulin which was originally isolated from BETULA or birch tree. It has anti-inflammatory, anti-HIV and antineoplastic activities.
See also: Paeonia lactiflora root (part of); Jujube fruit (part of).

C30H48O3

456.7

456.36

C, 78.90; H, 10.59; O, 10.51

472-15-1

64971

White to off-white solid powder

1.1±0.1 g/cm3

550.0±33.0 °C at 760 mmHg

295-298 °C (dec.)(lit.)

300.5±21.9 °C

0.0±3.4 mmHg at 25°C

1.533

8.94

57.53

2

3

2

33

861

10

O([H])[C@@]1([H])C([H])([H])C([H])([H])[C@@]2(C([H])([H])[H])[C@]([H])(C1(C([H])([H])[H])C([H])([H])[H])C([H])([H])C([H])([H])[C@]1(C([H])([H])[H])[C@]2([H])C([H])([H])C([H])([H])[C@]2([H])[C@@]3([H])[C@]([H])(C(=C([H])[H])C([H])([H])[H])C([H])([H])C([H])([H])[C@]3(C(=O)O[H])C([H])([H])C([H])([H])[C@@]12C([H])([H])[H]

QGJZLNKBHJESQX-FZFNOLFKSA-N

InChI=1S/C30H48O3/c1-18(2)19-10-15-30(25(32)33)17-16-28(6)20(24(19)30)8-9-22-27(5)13-12-23(31)26(3,4)21(27)11-14-29(22,28)7/h19-24,31H,1,8-17H2,2-7H3,(H,32,33)/t19-,20+,21-,22+,23-,24+,27-,28+,29+,30-/m0/s1

(1R,3aS,5aR,5bR,7aR,9S,11aR,11bR,13aR,13bR)-9-hydroxy-5a,5b,8,8,11a-pentamethyl-1-prop-1-en-2-yl-1,2,3,4,5,6,7,7a,9,10,11,11b,12,13,13a,13b-hexadecahydrocyclopenta[a]chrysene-3a-carboxylic acid

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 requires protection from light (avoid light exposure) during transportation and storage.

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

DMSO: 20~35.7 mg/mL (43.8~78.2 mM)
Ethanol: 10 mg/mL (~21.9 mM)

配方 1 中的溶解度: ≥ 2.08 mg/mL (4.55 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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配方 2 中的溶解度: 4 mg/mL (8.76 mM) in 50% PEG300 50% Saline (这些助溶剂从左到右依次添加，逐一添加), 悬浊液; 超声助溶。
*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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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;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
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。