3-Aminopropylphosphinic acid (3-APPA; CGP 27492; CGA 147823)   中文名称: 3-氨基丙基膦酸

GABAB receptor

3-氨基丙基次膦酸（10 μM）可作为皮肤抗衰老药物[4]。它还会诱导电刺激回肠中胆碱能抽搐收缩的浓度依赖性抑制（IC50=1.84-0.23 μM）[2]。

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测量了GABA、β-丙氨酸和甘氨酸的膦类似物对豚鼠回肠纵肌的影响。3-氨基丙基次膦酸（AMPh）和2-氨基乙基膦酸（2-AEPh）在非刺激制剂和电刺激制剂中均无任何作用。GABA的膦类似物3-Aminopropylphosphinic acid/3-氨基丙基膦酸（3-APPh）在10（-3）M的剂量下具有GABAB激动作用（放松和抑制抽搐反应）。没有观察到对GABAA受体的激动作用。3-APPh在测试剂量（2 X 10（-4）M和10（-3）M）下也对GABAB激动剂的作用显示出拮抗作用，导致GABA和（-）-巴氯芬抑制抽搐反应的对数剂量-效应曲线平行偏移。相比之下，3-APPh没有拮抗吗啡和去甲肾上腺素的抑制作用。通过GABAA受体介导的GABA的收缩作用不受3-APPh（10（-3）M）的影响。结论3-APPh是豚鼠回肠GABAB位点的部分激动剂。[1]

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1.测试了γ-氨基丁酸（GABA）类似物3-氨基丙基次膦酸对豚鼠离体回肠和大鼠离体肛尾肌制剂的活性。比较了3-Aminopropylphosphinic acid/3-氨基丙基次膦酸与GABA和巴氯芬的作用。2.在电刺激的回肠中，3-氨基丙基次膦酸，如GABA和巴氯芬，对胆碱能抽搐收缩产生浓度依赖性抑制，IC50值为1.84+/-0.23 microM（n=12）。与GABA不同，但与巴氯芬一样，3-氨基丙基次膦酸不会产生初始收缩。3.3-氨基丙基次膦酸和巴氯芬在豚鼠回肠中的抑制作用未被荷包牡丹碱（10微M）、酚妥拉明加普萘洛尔（均为1微M），育亨宾（1微M。然而，3-氨基丙基次膦酸的抑制作用，而巴氯芬的抑制作用则没有，被phaclofen（500微M）拮抗。此外，巴氯芬脱敏可消除豚鼠回肠中3-氨基丙基次膦酸的影响。4.3-氨基丙基次膦酸、GABA和巴氯芬可减少电场刺激引起的大鼠尾骨肌抽搐收缩。3-氨基丙基次膦酸抑制尾骨收缩的IC50为0.89+/-0.15微M（n=8）。5.结论是，3-氨基丙基次膦酸是一种强效、选择性的GABAB激动剂，在豚鼠回肠中的效力是巴氯芬的七倍，在大鼠肛门括约肌制剂中的效力比巴氯芬强五倍。[2]

3-氨基丙基次膦酸（5 mg/kg；静脉注射）通过阻断 GABA 的作用来抑制豚鼠的迷走神经支气管痉挛[3]。

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GABA是中枢神经系统中一种已知的抑制性神经递质。最近的研究还表明，GABA存在于包括肺在内的外周组织中。为了阐明GABA在肺中的作用，研究了GABA和选择性GABA激动剂和拮抗剂对豚鼠神经元诱导的气道收缩的影响。在体外，河豚毒素和阿托品抑制了电场刺激（EFS）诱导的气管收缩，表明收缩是由乙酰胆碱的神经元释放介导的。由EFS引起的收缩，而不是由外源性乙酰胆碱引起的收缩被GABA（EC50=4.5微M）和选择性GABA-B激动剂巴氯芬（EC50=9微摩）抑制，但不被GABA-A激动剂麝香醇抑制。巴氯芬的抑制作用不受GABA-A拮抗剂荷包牡丹碱的影响，但被GABA-B拮抗剂3-Aminopropylphosphinic acid/3-氨基丙基次膦酸（3-APPA）（pA2=4.5）和2-羟基乙酰氯芬（pA2=4.1）显著逆转。在体内，麻醉、机械通气豚鼠的迷走神经刺激（5 V，20 Hz，0.5 ms，5 s）引起胆碱能依赖性支气管痉挛，静脉注射GABA（3和10 mg/kg）和巴氯芬（1-10mg/kg）可抑制该痉挛，但麝香醇不能抑制。GABA和巴氯芬对迷走神经支气管痉挛的抑制作用被3-APPA（5mg/kg，静脉注射）阻断，但未被荷包牡丹碱阻断。分别用酚妥拉明或普萘洛尔阻断α肾上腺素能受体和β肾上腺素能受体治疗动物后，对GABA-B激动剂的反应没有改变。GABA和巴氯芬也不会改变静脉注射乙酰甲胆碱引起的支气管痉挛[3]。

“体外”制剂[1]
实验在雄性豚鼠（体重范围300-500g）中进行；动物们被头部的一击所伤；快速去除末端回肠的片段，并将其放置在以下成分（mM）的改良克雷布斯溶液中：KHzP041.3，KCI 3.4，NaCl134.7，CaC12 2.8，MgSO4 0.6，NaHC03 16.3，葡萄糖7.7。通过Paton和Zar（1968）的方法获得了附着有肌间神经丛的回肠纵向肌条。将管段安装在器官浴中，用5%COZ和95%02的混合物起泡，并保持在37°C。必要时，按照Paton（1963）和Paton&VLzi（1969）描述的方法进行电刺激。使用MARB刺激器的两个同轴铂电极施加刺激（持续时间为1msec的最大矩形脉冲的1.5倍，频率为每分钟6次）。在0.5g的静张力下将回肠连接到等距换能器，并在MARB多导生理记录仪上记录反应。给药前，让制剂平衡60分钟。给药量不得超过总浴量的1%（4毫升）。通过在两次给药之间间隔20分钟，可以防止GABA脱敏的发生。事实上，正如我们之前观察到的（Giotti等人，1983a；b），在非刺激和电刺激制剂中，以15-30min的间隔重复亚最大剂量的GABA都会引起相同的效果。

Animal/Disease Models: guinea pigs[3]
Doses: 5 mg/kg
Route of Administration: IV
Experimental Results: Blocked the inhibitory effects of GABA against vagal broncho-spasm.

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Guinea-pig ileum [2]
Male guinea-pigs (300-450g) were killed by a blow to the head and bled out. Segments of ileum of approximately 3 cm in length were removed from an area 10-15cm proximal to the ileo-caecal junction. Preparations were immediately placed in a modified Krebs solution (BUlbring, 1953) which was continually bubbled with 95% 02 and 5% CO2. Segments were freed of their mesenteric attachment and suspended in 10 ml organ baths containing Krebs solution under an isometric tension of 1 g. Isometric contractions to transmural stimulation (Paton, 1954) were recorded with strain gauge transducers and displayed on an Ormed Multitrace pen recorder. Electrical stimulation of preparations was achieved by passing rectangular pulses (duration 0.5 ms; frequency 0.1 Hz; supramaximal voltage (25-35 V)), from a Grass SD11 stimulator via platinum electrodes. Preparations were allowed to equilibrate for 1 h prior to addition of compounds to the organ bath.

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Rat anococcygeus [2]
Male Wistar rats (200-300 g) were killed by a blow to the head and bled out and their anococcygeus muscles removed as previously described (Gillespie, 1972). The muscles were mounted in organ baths (10 ml) containing modified Krebs solution which was continually gassed with 95% 02 and 5% CO2. A resting tension of 0.5 g was applied and the preparations were field stimulated from Grass SD11 stimulators via platinum ring electrodes with the following stimulus parameters: pulse duration 1 ms; frequency 10 Hz; for 1 s. Isometric muscle responses were measured with a strain gauge transducer and displayed on an Ormed Multitrace pen recorder. Typical tension responses generated in either preparation as a result of electrical stimulation were between 2 and 4 g force. Preparations generating less tension were rejected. In both preparations, sequential agonist concentration-response curves were constructed, allowing 30 min between additions of agonist to minimize tachyphylaxis. When antagonists were used, concentration-response curves to 3-Aminopropylphosphinic acid in the presence of the antagonist were constructed after an initial equiliThe following drugs were used: y-amino-n-butyric acid, (±+baclofen, (±)-propranolol (ICI), phentolamine mesylate, yohimbine hydrochloride, naloxone hydrochloride, 8-phenyltheophylline, impromidine oxylate, bicuculline methiodide, phaclofen and 3-Aminopropylphosphinic acid (prepared by the method of Dingwall et al., 1987a, b). With the exception of 8-phenyltheophylline, all compounds were dissolved in distilled water, subsequent dilutions being made in distilled water and compounds were added to the organ bath in volumes no greater than 1% total volume. 8- Phenyltheophylline was made up in 80% methanol/ 2 M NaOH, subsequent dilutions being made in distilled water and all vehicle controls were negative. Propranolol and phentolamine were added directly to the Krebs solution.

[1]. GABA-related activities of amino phosphonic acids on guinea-pig ileum longitudinal muscle. J Auton Pharmacol. 1986 Sep;6(3):163-9.
[2]. 3-Aminopropylphosphinic acid--a potent, selective GABAB receptor agonist in the guinea-pig ileum and rat anococcygeus muscle. Br J Pharmacol. 1989 Aug;97(4):1292-6.
[3]. Prejunctional GABA-B inhibition of cholinergic, neurally-mediated airway contractions in guinea-pigs. Pulm Pharmacol. 1991;4(4):218-24.
[4]. 3-Aminopropyl dihydrogen phosphate (3-APPA; 3-aminopropane phosphoric acid); a novel anti-aging substance. Journal of Investigative Dermatology, vol. 4, no. 106, 2015, p. 895.

Bioulac, De Tinguy-Moreaud, Vincent, and Neuzil (1979) described the inhibitory effect of 3-APPh on central neuronal firing, an effect insensitive to dicyclopentene (possibly a GABAB receptor); furthermore, Cates, Li, Yakashe, et al. (1984) recently confirmed that this compound has a certain affinity for the GABA binding site. Regarding GABAA receptors, none of the tested phosphonates showed agonistic activity against this subtype: only 3-APPh caused ileal contraction, but this contraction was insensitive to dicyclopentene and bitter substances, and GABA could not desensitize this contraction. The ineffectiveness of 3-APPh against GABAE receptors is also consistent with previous binding studies at the central level (Galli, Zilletti, Scotton, Adembri, and Giotti, 1980; Cates et al., 1984). However, our primary focus was on investigating the antagonistic effects of these drugs on GABA receptors: none showed interference with GABAA receptor-mediated contraction; conversely, high doses of 3-APPh exhibited significant antagonism against GABAB-mediated inhibition; the antagonism of 3-APPh was reversible and specific to GABAergic drugs. 2-AEPh and 3-APPh were ineffective. One question arising from this is whether the agonistic effect of 3-APPh on GABAB receptors interferes with its antagonistic effect on the same receptor through desensitization. The fact that 3-APPh rapidly reverses the effect of GABAA on muscle twitching (Figure 6) supports the idea of a direct antagonistic effect. Furthermore, at lower test doses (2 × 10⁻⁵ M), the antagonistic effect of 3-APPh on the GABAB effect appeared competitive: the dose-response curves shifted parallel, with little change in the maximum effect. On the other hand, desensitization may occur at the highest dose (10⁻³ M); in fact, at this concentration, the dose-response curve tends to flatten, which may be due to desensitization. In summary, the GABA phosphonate analog 3-aminopropylphosphonic acid (3-APPh) exhibits characteristics of both a weak GABA_B receptor antagonist and a weak GABA_B receptor agonist, while having no effect on GABA_A receptors (agonist or antagonist). This characteristic may be interpreted as partial agonism, unlike other drugs considered GABA_A receptor antagonists (such as 3-APS, which has characteristics of both a strong GABA_A receptor agonist and a weak GABA_B receptor antagonist) (Table 2). Therefore, it appears that no effective GABA_B receptor antagonist has yet been discovered. However, a precise understanding of the differences between drugs acting on GABAB receptors can help in using them as experimental tools and in developing more selective GABAB antagonists. [1] Several studies have explored the requirements for GABAB receptor activity, showing that even small changes to the baclofen molecule can lead to a complete loss of its activity (Olpe et al., 1980; Krogsgard-Larsen, 1988). To date, no GABAB agonist has been found to be more effective than baclofen in the in vitro systems described herein. 3-aminopropylphosphonic acid is 7 times more potent than racemic baclofen in the guinea pig ileum and 5 times more potent than racemic baclofen in the rat anal and caudal muscles. Studies reported by Dingwall et al. (1987a, b) have shown that 3-aminopropylphosphonic acid has a 20-fold greater affinity for GABAB receptors than baclofen. Interestingly, a comparison is made between GABA phosphonate analogs described in this paper as potent agonists and those described as weak partial agonists/antagonists (Luzzi et al., 1986), despite the only structural difference being the acidic moiety. 3-Aminopropylphosphonic acid has a distorted tetrahedral arrangement around its phosphorus atom, contains only one acidic proton, and has a negative charge distributed across the two oxygen atoms, thus making it similar to GABA in many respects. 3-Aminopropylphosphonic acid has a nearly tetrahedral arrangement around its phosphorus atom, contains two acidic protons (pH-dependent), and has a negative charge distributed across the three oxygen atoms. In guinea pig ileum, 3-aminopropylphosphonic acid appears to interact with presynaptic GABA receptors located at cholinergic nerve endings. Since the Sid value of 3-aminopropylphosphonic acid is not significant and it does not induce initial contraction, it may not possess GABAA receptor agonist activity. Although a more specific O value is needed for final statistical validation of this hypothesis, the results using the known specific receptor antagonist Iic acidm(l) indicate that 3-aminopropylphosphonic acid does not interact with any other type of receptor. Furthermore, tissues desensitized to baclofen no longer respond to 3-aminopropylphosphonic acid. Clonidine (with a mean above average, p < 0.05) was used to test the specificity of GABAB receptors. Clonidine showed comparable potency to baclofen during GABAB receptor desensitization, with significant differences before and after desensitization. Faclofen is considered a weak but selective GABAB antagonist (Kerr et al., 1987; Dutar and Nicholl, 1988). We were able to demonstrate that the response to 3-aminopropylphosphonic acid was inhibited in guinea pig ileum, but the response to baclofen was not inhibited. The reason is currently unclear, although recent studies have claimed that faclofen exhibits GABA antagonistic activity in multiple testing systems (Karlsson et al., 1988; Soltesz et al., 1988), but its structure does support our hypothesis that the 3-aminopropylphosphonic acid (MPA) interacts with the GABA receptor. [2]
In guinea pig ileum and rat anal-coccygeal muscle, the response time course of MPA was similar to that of GABA and baclofen. In guinea pig ileum, the response to the GABAB receptor agonist was generally transient, while in rat anal-coccygeal muscle, the response was more persistent (Bowery et al., 1981; Muhyaddin et al., 1982). Our pharmacological studies support the view of Dingwall et al. (1987a, b) that MPA is a potent and selective GABAB receptor agonist. Further research on this compound may help elucidate the physiological role of the GABAB receptor in the mammalian gut. [2]

C3H10NO2P

122.08286

122.037

103680-47-3

6335948

Typically exists as solid at room temperature

0.001mmHg at 25°C

-1.4

95.92

2

3

3

7

66

0

NCCCP(=O)O

MQIWYGZSHIXQIU-UHFFFAOYSA-O

InChI=1S/C3H8NO2P/c4-2-1-3-7(5)6/h1-4H2/p+1

3-aminopropyl-hydroxy-oxophosphanium

3-aminopropylphosphinic acid; 103680-47-3; 3-aminopropyl-hydroxy-oxophosphanium; 3-Aminopropanephosphinic acid; (3-aminopropyl)phosphinic acid; CGP-27492; 3-APPA; Cgp 27492;

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));
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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；
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