Farnesyl diphosphate synthase (IC50 = 460 nM)

阿仑膦酸钠水合物直接作用于破骨细胞时，会抑制破骨细胞功能所必需的磷脂产生的限速阶段[1]。 Alendronate Sodium Hydrate 剂量依赖性地抑制 [3H]MVA 对甾醇的摄取，同时 IPP 和 DMAPP 的剂量标志物增加 [3]。阿仑膦酸钠水合物通过降低香叶基香叶基二磷酸（一种 18-25 kDa 的蛋白质和非皂化抑制剂，包括破骨细胞中的甾醇）的水平来抑制 [3H]甲瓦龙酸内酯 [2]。

在兔子中，阿仑膦酸钠水合物会导致胃糜烂，但不会导致胃窦发育。阿仑膦酸钠水合物可增加由卡美辛引起的胃窦的频率和幅度。阿仑膦酸钠水合物会延迟胃愈合，并放大碳水化合物胡萝卜素的作用[4]。当与紫杉醇（10-50 mg/kg/天，每天两次或两次）联合使用时，阿仑膦酸钠水合物（0.04-0.1 mg/kg，每周两次或每周 0.1 mg/kg）部分抑制人 PC-3 ML 的骨转移用它处理的小鼠的细胞。这可导致PC-3 ML在骨髓和软组织中致命性爆发肿瘤生长，但其发生率在4-5周内显着提高[5]。

阿仑膦酸盐是一种含氮的双膦酸盐，是一种强效的骨吸收抑制剂，用于治疗和预防骨质疏松症。最近的研究结果表明，阿仑膦酸盐和其他含氮双膦酸盐抑制类异戊二烯生物合成途径，并干扰蛋白质异戊二烯化，这是由于香叶基香叶基二磷酸水平降低的结果。本研究确定法尼醇二磷酸合酶是双膦酸盐抑制的甲羟戊酸途径酶。对肝脏细胞质提取物产物的HPLC分析将阿仑膦酸盐抑制的潜在靶点（IC（50）=1700 nM）缩小到异戊烯基二磷酸异构酶和法尼基二磷酸合酶。阿仑膦酸盐抑制重组人法尼烷基二磷酸合酶，IC（50）为460 nM（预孵育15分钟后）。阿仑膦酸盐不抑制异戊烯基二磷酸异构酶或GGPP合酶，部分从肝细胞质中纯化。重组法尼烷基二磷酸合酶也被帕米膦酸二钠（IC（50）=500 nM）和利塞膦酸二钾（IC（50中）=3.9 nM）抑制，被依替膦酸二铵可忽略（IC50=80微M），而完全不被氯膦酸二钙抑制。在破骨细胞中，阿仑膦酸盐抑制了[（3）H]甲羟戊酸内酯掺入18-25 kDa的蛋白质和非脂质体，包括甾醇。这些发现（i）确定法呢酰二磷酸合酶是甲羟戊酸途径中阿仑膦酸盐的选择性靶标，（ii）表明该酶被其他含氮双膦酸盐抑制，如利塞膦酸盐，但不被氯膦酸盐抑制，支持不同双膦酸盐的不同作用机制，以及（iii）在纯化的破骨细胞中记录了阿仑膦酸酯对异戊二烯化和甾醇生物合成的抑制作用。[2]

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阿仑膦酸盐（ALN）是一种用于治疗骨质疏松症和其他骨吸收疾病的氨基双膦酸盐化合物，已被认为通过抑制GGPP的形成起作用。在本研究中，我们使用大鼠肝脏的S（10）匀浆部分来证明，ALN对[（3）H]MVA掺入甾醇具有剂量依赖性抑制作用，并伴随着放射性标记掺入IPP和DMAPP的增加。我们进一步表明，ALN是细胞质反式异戊二烯基转移酶（FPP合酶）的强效抑制剂。这种抑制作用与烯丙基焦磷酸盐底物具有竞争性，但与IPP没有竞争性，这表明ALN充当烯丙基焦磷酸类似物并与游离酶结合。K（i）在0.5微米范围内。

含氮双膦酸盐被证明可以通过抑制从甲羟戊酸到胆固醇的生物合成途径中的酶来引起巨噬细胞凋亡。这项研究表明，在破骨细胞中，香叶基香叶基二磷酸（大多数GTP结合蛋白的异戊二烯化底物）可能是受这些双膦酸盐影响的关键中间体。我们报告说，洛伐他汀（羟甲基戊二酰辅酶A还原酶抑制剂）和阿仑膦酸盐都能抑制培养中的小鼠破骨细胞形成。洛伐他汀的作用被甲羟戊酸完全阻断，而香叶基香叶醇的阻断效果较差，而阿仑膦酸钠的作用被甲羟戊酸部分阻断，被香叶基香叶醇更有效。阿仑膦酸钠对小鼠颅盖骨吸收的抑制作用也被甲羟戊酸阻断，而氯屈膦酸钠的抑制作用则没有。此外，洛伐他汀和阿仑膦酸钠也抑制了兔破骨细胞的形成和活性。甲羟戊酸或香叶基香叶醇可以阻止洛伐他汀的作用，而香叶基香叶醇可以防止阿仑膦酸盐的作用。法尼醇和角鲨烯没有效果。信号研究表明，洛伐他汀和阿仑膦酸盐在纯化的破骨细胞中激活了一种34kDa的激酶。洛伐他汀介导的激活被甲羟戊酸和香叶基香叶醇阻断，而阿仑膦酸钠的激活被香叶基香叶醇阻断。这些发现共同支持了阿仑膦酸盐直接作用于破骨细胞，抑制胆固醇生物合成途径中对破骨细胞功能至关重要的限速步骤的假设。外源性香叶基香叶醇可以阻止这种抑制作用，这可能是控制细胞骨架重组、囊泡融合和凋亡的GTP结合蛋白异戊二烯化所必需的，这些过程涉及破骨细胞的活化和存活[1]。

Gastric ulceration associated with the use of NSAIDs is most frequently observed in elderly women, the same sector of society most likely to be receiving therapy for osteoporosis. As some anti-osteoporosis medications have been suggested to irritate the upper gastrointestinal mucosa, we evaluated the ability of one such drug, alendronate, to damage the gastric mucosa and to influence the severity and healing of gastric ulcers in rodents. The effects of alendronate on indomethacin-induced antral ulceration was evaluated in the rabbit, while effects on ulcer healing and on the formation of gastric erosions was evaluated in the rat. Effects of alendronate on gastric acid secretion, blood flow and prostaglandin synthesis were also evaluated. Alendronate caused erosions in the rabbit stomach, but not antral ulceration. However, at the highest doses tested (80 mg) alendronate increased the incidence and size of indomethacin-induced antral ulcers. Alendronate also enhanced indomethacin-induced gastric damage in the rat, and delayed gastric ulcer healing. These effects of alendronate were not attributable to changes in gastric acid secretion, blood flow, prostaglandin synthesis or the pharmacokinetics of indomethacin. The damaging effects of alendronate on the stomach were due to topical irritant effects and could be observed at concentrations as low as 4 mg/ml within 30 min of oral administration or topical superfusion. These results support preliminary clinical evidence that alendronate can damage the gastric mucosa. While gastric injury may be a rare occurrence in patients taking this drug, concomitant use of alendronate and NSAIDs may increase the incidence or severity of ulceration.[4]

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The combined influence of alendronate, a bisphosphonate compound, and taxol on the establishment and growth of human PC-3 ML subclones injected intravenously via the tail vein in SCID mice was investigated. The pretreatment of SCID mice with alendronate (0.04-0.1 mg/kg twice weekly or 0.1 mg/kg weekly) partially blocked the establishment of bone metastases by human PC-3 ML cells and resulted in tumor formation in the peritoneum and other soft tissues. However, alendronate pretreatment of mice (0.1 mg/kg twice weekly or weekly) and dosing along with taxol (10-50 mg/kg/day, twice weekly, or weekly) blocked the growth of PC-3 ML tumors in the bone marrow and soft tissues in a statistically significant manner and improved survival rates significantly (p < 0.001) by 4-5 weeks. ELISAs and zymography of matrix metalloproteinase production in vitro and in vivo showed that alendronate and taxol alone partially inhibited metalloproteinase production, but that taxol in combination with alendronate totally blocked protease production and release. The combined activities of alendronate and taxol appeared to inhibit the establishment and growth of tumors in SCID mice, perhaps, in part, as a result of inhibition of protease production and release.[5]

Effects During Pregnancy and Lactation
◉ Overview of Medication Use During Lactation
Limited evidence suggests that discontinuing breastfeeding after long-term bisphosphonate treatment does not appear to have adverse effects on the infant. Since there is currently no information on the use of alendronate sodium during lactation, alternative medications may be preferred, especially for breastfed newborns or premature infants. However, breastfed infants are unlikely to absorb alendronate sodium. If the mother takes bisphosphonates during pregnancy or lactation, some experts recommend monitoring the infant's serum calcium levels in the first two months postpartum. ◉ Effects on Breastfed Infants
Because alendronate sodium can persist in the body for several years after long-term use, the following case may be relevant. A woman took alendronate sodium for 6 months within a year prior to conception, followed by pamidronate sodium every 4 months. Her infant was breastfed for 3 months (feeding extent not specified). The infant developed mild hypocalcemia at 2 months of age, but calcium levels were normal at 5 months of age, and long bone development was normal.
◉ Impact on breastfeeding and breast milk
As of the revision date, no relevant published information was found.

[1]. Fisher JE, et al. Alendronate mechanism of action: geranylgeraniol, an intermediate in the mevalonate pathway, prevents inhibition of osteoclast formation, bone resorption, and kinase activation in vitro. Proc Natl Acad Sci U S A. 1999 Jan 5;96(1):133-8.
[2]. Bergstrom JD, et al. Alendronate is a specific, nanomolar inhibitor of farnesyl diphosphate synthase. Arch Biochem Biophys. 2000 Jan 1;373(1):231-41.
[3]. Keller RK, et al. Mechanism of aminobisphosphonate action: characterization of alendronate inhibition of the isoprenoid pathway. Biochem Biophys Res Commun. 1999 Dec 20;266(2):560-3.
[4]. Elliott SN, et al. Alendronate induces gastric injury and delays ulcer healing in rodents. Life Sci. 1998;62(1):77-91.
[5]. Stearns ME, et al. Effects of alendronate and taxol on PC-3 ML cell bone metastases in SCID mice. Invasion Metastasis. 1996;16(3):116-31

Alendronate sodium is the sodium salt of alendronate, a second-generation bisphosphonate and a synthetic analog of pyrophosphate with anti-bone resorption activity. Alendronate sodium binds to and inhibits the activity of geranyltransferase (farnesyl pyrophosphate synthase), an enzyme involved in the biosynthesis of terpenoids. Inhibition of this enzyme prevents the biosynthesis of isoprene lipids (FPP and GGPP), which are donor substrates for farnesylation and geranyl-geranylation during the post-translational modification of small GTPase signaling proteins, modifications crucial for osteoclast turnover. Therefore, osteoclast activity is inhibited, leading to reduced bone resorption and turnover.
A non-hormonal drug used to treat osteoporosis in postmenopausal women. This drug promotes healthy bone growth and restores some of the bone loss caused by osteoporosis.
See also: Alendronate sodium (note moved here).

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Indications
For the treatment of postmenopausal osteoporosis, especially in patients at risk of vitamin D deficiency. Adrovance can reduce the risk of vertebral and hip fractures.

C4H12NNAO7P2

271.08

270.998

129318-43-0

Alendronate sodium hydrate;121268-17-5;Alendronic acid;66376-36-1

16760285

Typically exists as solid at room temperature

616.7ºC at 760 mmHg

326.7ºC

180.93

5

8

5

15

293

0

C(CC(O)(P(=O)(O)O)P(=O)(O)[O-])CN.[Na+]

CAKRAHQRJGUPIG-UHFFFAOYSA-M

InChI=1S/C4H13NO7P2.Na/c5-3-1-2-4(6,13(7,8)9)14(10,11)12;/h6H,1-3,5H2,(H2,7,8,9)(H2,10,11,12);/q;+1/p-1

sodium;(4-amino-1-hydroxy-1-phosphonobutyl)-hydroxyphosphinate

ALENDRONATE SODIUM; 129318-43-0; Fosamax; Binosto; Fosamac; Onclast; alendronate monosodium; Monosodium alendronate;

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)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

注意: 如下所列的是一些常用的体内动物实验溶解配方，主要用于溶解难溶或不溶于水的产品（水溶度<1 mg/mL）。 建议您先取少量样品进行尝试，如该配方可行，再根据实验需求增加样品量。

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注射用配方1: DMSO : Tween 80： Saline = 10 : 5 : 85 (如： 100 μL DMSO → 50 μL Tween 80 → 850 μL Saline)
*生理盐水/Saline的制备：将0.9g氯化钠/NaCl溶解在100 mL ddH ₂ O中，得到澄清溶液。
注射用配方 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (如： 100 μL DMSO → 400 μL PEG300 → 50 μL Tween 80 → 450 μL Saline)
注射用配方 3: DMSO : Corn oil = 10 : 90 (如： 100 μL DMSO → 900 μL Corn oil)
示例: 以注射用配方 3 (DMSO : Corn oil = 10 : 90) 为例说明, 如果要配制 1 mL 2.5 mg/mL的工作液, 您可以取 100 μL 25 mg/mL 澄清的 DMSO 储备液，加到 900 μL Corn oil/玉米油中, 混合均匀。

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注射用配方 4: DMSO : 20% SBE-β-CD in Saline = 10 : 90 [如：100 μL DMSO → 900 μL (20% SBE-β-CD in Saline)]
*20% SBE-β-CD in Saline的制备（4°C，储存1周）：将2g SBE-β-CD (磺丁基-β-环糊精) 溶解于10mL生理盐水中，得到澄清溶液。
注射用配方 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (如： 500 μL 2-Hydroxypropyl-β-cyclodextrin (羟丙基环胡精)→ 500 μL Saline)
注射用配方 6: DMSO : PEG300 : Castor oil : Saline = 5 : 10 : 20 : 65 (如： 50 μL DMSO → 100 μL PEG300 → 200 μL Castor oil → 650 μL Saline)
注射用配方 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (如： 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
注射用配方 8: 溶解于Cremophor/Ethanol (50 : 50), 然后用生理盐水稀释。
注射用配方 9: EtOH : Corn oil = 10 : 90 (如： 100 μL EtOH → 900 μL Corn oil)
注射用配方 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (如： 100 μL EtOH → 400 μL PEG300 → 50 μL Tween 80 → 450 μL Saline)

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口服配方 1: 悬浮于0.5% CMC Na (羧甲基纤维素钠)
口服配方 2: 悬浮于0.5% Carboxymethyl cellulose (羧甲基纤维素)
示例: 以口服配方 1 (悬浮于 0.5% CMC Na)为例说明, 如果要配制 100 mL 2.5 mg/mL 的工作液, 您可以先取0.5g CMC Na并将其溶解于100mL ddH2O中，得到0.5%CMC-Na澄清溶液；然后将250 mg待测化合物加到100 mL前述 0.5%CMC Na溶液中，得到悬浮液。

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口服配方 3: 溶解于 PEG400 (聚乙二醇400)
口服配方 4: 悬浮于0.2% Carboxymethyl cellulose (羧甲基纤维素)
口服配方 5: 溶解于0.25% Tween 80 and 0.5% Carboxymethyl cellulose (羧甲基纤维素)
口服配方 6: 做成粉末与食物混合

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注意: 以上为较为常见方法，仅供参考， InvivoChem并未独立验证这些配方的准确性。具体溶剂的选择首先应参照文献已报道溶解方法、配方或剂型，对于某些尚未有文献报道溶解方法的化合物，需通过前期实验来确定（建议先取少量样品进行尝试），包括产品的溶解情况、梯度设置、动物的耐受性等。

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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。