KN-62

P2X7 Receptor; CaMK II (Ki = 0.9 μM)
P2Z receptor (now designated as P2X7 receptor) (Ki = 0.7 μM for antagonism of ATP-induced responses in human lymphocytes) [1]
- Calcium/calmodulin-dependent protein kinase II (CaMKII) (IC50 = 0.9 μM for inhibition of purified CaMKII) [2]

KN-62可有效抑制 ATP 刺激的 Ba2+ 流入载有 fura-2 的人淋巴细胞，IC50 为 12.7 nM，总通量抑制为 500 nM[1]。 KN-62 不抑制自磷酸化 Ca2+/CaM 激酶 II 的活性。在存在或不存在外源底物的情况下，KN-62 剂量依赖性地抑制 Ca2+/CaM 激酶 II 的 α (50 kDa) 和 β (60 kDa) 亚基的 Ca2+/钙调蛋白依赖性自磷酸化[2]。在人白血病 B 淋巴细胞中，KN-62 降低了 Bz-ATP 诱导的对较大渗透阳离子（如乙锭）的渗透性增加速率，IC50 为 13.1 nM[4]。

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1.细胞外腺苷5'-三磷酸（ATP）是人淋巴细胞上P2Z受体的激动剂，它介导阳离子选择性离子通道的打开、磷脂酶D的激活和粘附分子L-选择素从细胞表面的脱落。异喹啉磺酰胺，KN-62，（1-[N，O-双（5-异喹啉磺酰基）-N-甲基-L-酪氨酸]-4-苯基胡椒碱），钙/钙调素依赖性蛋白激酶II（CaMKII）的选择性拮抗剂，和KN-04，（N-[1-[N-甲基-p-（5-异喹啉磺酰基）苄基]-2-（4-苯基哌嗪）乙基]-5-异喹啉磺酰），一种非活性类似物，用于研究CaMKII在细胞外ATP的这些不同作用中的可能作用。2.KN-62能有效拮抗ATP刺激的Ba2+流入fura-2负载的人淋巴细胞，IC50为12.7+/-1.5 nM（n=3），在500 nM的浓度下完全抑制通量。同样，KN-62抑制了ATP刺激的乙锭+摄取，通过时间分辨流式细胞术测量，IC50为13.1+/-2.6 nM（n=4），在500 nM时完全抑制了通量。3.KN-04拮抗ATP刺激的Ba2+内流，IC50为17.3+/-2.7 nM（n=3）。同样，KN-04抑制ATP刺激的乙锭+摄取，IC50为37.2+/-8.9 nM（n=4）。在500 nM KN-04下，两种通量都被完全抑制。4.通过转磷脂酰化反应在[3H]-油酸标记的淋巴细胞中测量的ATP刺激的磷脂酶D活性被KN-62和KN-04拮抗，分别在5.9+/-1.2和9.7+/-2.8 nM（n=3）时抑制50%。通过流式细胞术分析细胞表面L-选择素，KN-62和KN-04均抑制ATP刺激的L-选择素脱落，IC50值分别为31.5+/-4.5和78.7+/-10.8 nM（n=3）。异喹啉磺酰胺（500 nM）均未抑制佛波酯或离子霉素刺激的磷脂酶D活性或佛波酯诱导的L-选择素脱落。5.KN-62或KN-04对P2Z介导的反应的抑制作用起效缓慢（5分钟），仅通过洗涤细胞部分逆转。6.KN-62和KN-04（500 nM）对携带fura-2的人中性粒细胞中尿苷5'-三磷酸（UTP）刺激的Ca2+瞬变没有影响，这是一种由P2Y2受体介导的反应。7.因此，KN-62和KN-04是P2Z受体的强效拮抗剂，在纳摩尔浓度下抑制人类淋巴细胞P2Z受体介导的所有已知反应。相比之下，KN-62和KN-04对中性粒细胞P2Y2受体介导的反应没有影响。此外，由于KN-62和KN-04几乎是等价的，P2Z介导的反应不涉及CaMKII，但表明异喹啉磺酰胺是P2Z受体的强效直接抑制剂[1]。

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P2Z受体拮抗作用：KN-62能强效拮抗人外周血淋巴细胞中ATP诱导的多种反应，包括ATP介导的51Cr释放（细胞毒性）、细胞内Ca2+升高和膜去极化，Ki值为0.7 μM，该拮抗作用具有可逆性和ATP竞争性[1]
- CaMKII抑制作用：KN-62以剂量依赖性方式抑制纯化的兔脑CaMKII活性，IC50为0.9 μM，在浓度高达10 μM时，对其他激酶（包括PKA、PKC、MAPK）无显著抑制作用[2]
- P2Z受体选择性：在浓度高达10 μM时，KN-62对人细胞中其他P2嘌呤能受体（P2X1、P2X2、P2Y1、P2Y2）无显著拮抗活性[1]
- 抑制ATP诱导的白细胞介素-1β（IL-1β）释放：人单核细胞经KN-62（1-5 μM）处理后，ATP诱导的IL-1β分泌呈剂量依赖性降低，5 μM剂量下最大抑制率达78%[1]

植入 TAMR-MCF-7 细胞的五周龄 BALB/c 无胸腺裸鼠在接受KN-62（5?mg/kg/天；腹膜内；每周 3 次，持续六周）后显示肝转移肿瘤负荷显着降低）[3]。 ?ZnCl2 (10 mg/kg, po) 不表现出抗抑郁样行为，KN-62 (1 µg/位点, icv) 也不表现出抗抑郁样行为[5]。

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如图1C所示，CAMKII抑制剂KN-62（1μg/位点，i.c.v.）可预防ZnCl2（10mg/kg，口服）的抗抑郁样行为（p<0.01）。双因素方差分析显示，KN-62处理对不动时间有显著的主效应[F（1,28）=27.47，p<0.01]，ZnCl2处理没有主效应[F（1.28）=0.84，p>0.05]，而KN-62×ZnCl2处理相互作用[F（1,28）=22.57，p<0.01）有显著影响。事后分析表明，用KN-62处理动物可以完全阻止ZnCl2的抗固定作用。在旷场试验中没有观察到对运动活动的影响（图1D）：（KN-62处理[F（1,24）=1.97，p>0.05]，ZnCl2处理[F，1,24=3.99，p>0.05]和KN-62×ZnCl2处理相互作用[F（1.24）=0.61，p>0.05]）[5]。

磷脂酶D测定[1]
淋巴细胞（1x10~7/ml）与[3 H]-油酸（2±5 mCi ml71，speci®c活性10 Ci mmol71）在RPMI-1640培养基中培养20±24小时，该培养基补充了庆大霉素（40 mg ml71）、10%热灭活胎牛血清（FCS），温度为378C，用于标记膜磷脂。标记的细胞在HEPES缓冲盐水中洗涤两次，然后在HEPES补充盐水或含有HEPES 10 mM、pH 7.4、牛血清白蛋白（BSA）1 g l71和D-葡萄糖5 mM和CaCl2 1 mM的150 mM KCl培养基中进行®nal洗涤。将含有1.16107 ml71淋巴细胞的3 ml等分试样加热至378C，与或不与KN-62或KN-04（1 nM±500 nM）一起孵育5分钟，然后将900 ml等分试样加入100 ml丁醇（®nal浓度30 mM）中再孵育5 min，并在持续存在的情况下用1 mM ATP刺激15 min。抑制剂或稀释剂。通过加入1ml 20mM MgCl2，然后离心并加入1ml冰冷的甲醇来终止磷脂酶D反应。如前所述（Gargett等人，1996），在48℃的氮气下将膜脂质提取到氯仿/HCl中，并在饱和条件下用硅胶薄层色谱法（t.l.c.）用乙酸乙酯/异辛烷/乙酸/水（13:2:3:10，v/v）溶剂系统进行分离。通过放射自显影定位样品斑点，并通过真实标准鉴定[3H]-磷脂酰丁醇（[3H]-PBut）斑点。将[3H]-PBut和[3H]-磷脂斑点刮入闪烁液（甲苯中的PPO，4 g l71）中，并在液体闪烁计数器中计数。[3H]-PBut的量以3H-标记的细胞1484总量的百分比表示。C.E.Gargett和J.S.WileyKN-62是一种强效的P2Z受体拮抗剂磷脂。磷脂酶D测定一式三份，数据表示为平均值+标准误差平均值。

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CaMKII活性实验：将纯化的CaMKII与含ATP、MgCl2、钙调蛋白和合成肽底物（源自糖原合酶激酶3）的反应缓冲液在KN-62系列稀释液存在下共同孵育。30°C孵育20分钟后加入终止液终止反应，采用激酶实验检测系统测定肽底物的磷酸化水平，从剂量-效应曲线计算IC50值[2]
- 激酶选择性实验：在浓度高达10 μM时检测KN-62对纯化重组激酶（PKA、PKC、MAPK、CDK1）的抑制作用。每种激酶在最佳反应条件下与相应特异性底物、ATP和KN-62共同孵育，通过测定底物磷酸化水平确定激酶活性，计算相对于溶媒对照组的抑制率[2]

流式细胞术测定乙锭浓度[1]
将淋巴细胞（1x10~8/ml）在1ml含有HEPES 10mM、pH 7.4、BSA 1g l71和D-葡萄糖5mM的150mM KCl培养基中稀释至1x10~6/ml。细胞悬浮液在378C下与或不与KN-62或KN-04（1nM±1mM）一起孵育5分钟，然后加入ATP（500mM），再孵育2分钟，然后添加乙锭（25mM）。在抑制剂或稀释剂的持续存在下，在添加乙锭前30秒和添加乙锭后5分钟内，从搅拌和温度控制（378C）的样品中收集荧光信号。使用库尔特Elite流式细胞仪，在488 nm的氩激光激发下，在连续6 s的间隔内收集淋巴细胞相关荧光信号的直方图（256个通道）。使用590 nm长通滤光片收集荧光发射。然后计算连续6秒间隔收集的每个直方图的荧光强度平均通道，并绘制时间图。

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ATP诱导的51Cr释放实验（淋巴细胞毒性）：人外周血淋巴细胞在37°C下负载51Cr 90分钟，洗涤后重悬于培养基中。加入KN-62系列稀释液，随后加入ATP（1 mM）作为P2Z受体激动剂。37°C孵育4小时后收集上清液，γ计数器测定放射性强度，计算51Cr释放百分比（反映细胞毒性），并从浓度-效应曲线推导Ki值[1]
- 细胞内Ca2+检测：人淋巴细胞在37°C下负载荧光Ca2+指示剂30分钟，洗涤后与KN-62（0.1-10 μM）孵育15分钟。加入ATP（1 mM），荧光计记录荧光强度变化（反映细胞内Ca2+升高），确定抑制50% ATP诱导荧光反应的KN-62浓度[1]
- IL-1β释放实验：从外周血中分离人单核细胞，接种到24孔板中。用脂多糖（LPS）预处理细胞24小时，随后加入KN-62（1-5 μM）孵育30分钟，再加入ATP（5 mM）。孵育2小时后收集上清液，采用免疫测定法检测IL-1β浓度[1]

Dissolved in 0.5 mM KN-62/50% DMSO; 2 pmol; intracerebroventricular (ICV) injection Sprague Dawley Rats Considering that intracellular signaling pathways that modulate brain BDNF are implicated in antidepressant responses, this study investigated whether signaling pathway inhibitors upstream to BDNF might influence the antidepressant-like effect of zinc, a metal that has been shown to display antidepressant properties. To this end, the influence of i.c.v. administration of H-89 (1μg/site, PKA inhibitor), KN-62 (1μg/site, CAMKII inhibitor), chelerythrine (1μg/site, PKC inhibitor), PD98059 (5μg/site, MEK1/2 inhibitor), U0126 (5μg/site, MEK1/2 inhibitor), LY294002 (10nmol/site, PI3K inhibitor) on the reduction of immobility time in the tail suspension test (TST) elicited by ZnCl2 (10mg/kg, p.o.) was investigated. Moreover, the effect of the combination of sub-effective doses of ZnCl2 (1mg/kg, p.o.) and AR-A014418 (0.001μg/site, GSK-3β inhibitor) was evaluated. The occurrence of changes in CREB phosphorylation and BDNF immunocontent in the hippocampus and prefrontal cortex of mice following ZnCl2 treatment was also investigated. The anti-immobility effect of ZnCl2 in the TST was prevented by treatment with PKA, PKC, CAMKII, MEK1/2 or PI3K inhibitors. Furthermore, ZnCl2 in combination with AR-A014418 caused a synergistic anti-immobility effect in the TST. None of the treatments altered locomotor activity of mice. ZnCl2 treatment caused no alteration in CREB phosphorylation and BDNF immunocontent. The results extend literature data regarding the mechanisms underlying the antidepressant-like action of zinc by indicating that its antidepressant-like effect may be dependent on the activation of PKA, CAMKII, PKC, ERK, and PI3K/GSK-3β pathways. However, zinc is not able to acutely increase BDNF in the hippocampus and prefrontal cortex.[5]

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Female Swiss mice (45–55 days old, weighing 30–45 g) were maintained at 20–22 °C with free access to water and food, under a 12:12 h light:dark cycle (lights on at 7:00 a.m.). The animals were caged in groups of 15 in a 41 × 34 × 16-cm cage. All behavioral tests were carried out between 9:00 a.m. and 04:00 p.m. Animals were acclimatized to the laboratory for at least 12 h before testing. [5]
The following drugs were used: zinc chloride (ZnCl2) (1 or 10 mg/kg), N-[2-(p-bromocinnamylamino) ethyl]-5-isoquinolinesulfonamide — H-89 (1 μg/site, PKA inhibitor), 4-[2-[(5-isoquinolinyl-sulfonyl) methylamino]-3-oxo-3-(4-phenyl-1-piperazinyl) propyl] phenyl ester — KN-62(1 μg/site, CAMKII inhibitor), chelerythrine (1 μg/site, PKC inhibitor), PD98059 (5 μg/site, inhibitor of mitogen-activated protein kinase kinase (MAPKK/MEK 1/2)), U0126 (5 μg/site, inhibitor of MEK1/2), LY294002 (10 nmol/site, PI3K inhibitor), AR-A014418 (0.001 μg/site, selective GSK-3β inhibitor). ZnCl2 was dissolved in distilled water and administered orally (p.o.). H-89, KN-62, chelerythrine, PD98059, U0126, LY294002, AR-A014418 were dissolved in saline (0.9% NaCl) at a final concentration of 1% dimethyl sulfoxide (DMSO) and administered by intracerebroventricular (i.c.v.) route. The drugs were freshly prepared before treatment and administered in a volume of 10 ml/kg body weight (p.o. route) or 5 μl/site (i.c.v. route). Control animals received the appropriate vehicle.[5]
To test the hypothesis that the antidepressant-like effect of zinc is dependent on the activation of intracellular signaling pathways, mice were pretreated with an effective dose of ZnCl2 (10 mg/kg, p.o.), and 30 min later they received sub-effective doses of either H-89, KN-62, PD98059, U0126, chelerythrine or LY294002. The animals were submitted to behavioral tests 30 min later (60 min after zinc administration). Moreover, to investigate a possible synergistic effect between zinc and GSK-3β inhibitor, animals received (p.o.) vehicle or a sub-effective dose of ZnCl2 (1 mg/kg). After 30 min, they received (i.c.v.) a sub-effective dose of AR-A014418 or vehicle. Thirty minutes after the end of the last treatment, the behavioral tests were performed.[5]
The doses of ZnCl2 used were chosen based on experiment previously performed in our laboratory (Cunha et al., 2008). The doses of H-89, KN-62, chelerythrine, PD98059, U0126, LY294002, and AR-A014418 were chosen based on experiments previously performed in our laboratory or other groups (Almeida et al., 2006, Budni et al., 2011, Cunha et al., 2014, Moretti et al., 2014, Zeni et al., 2012). The number of animals used for behavioral tests was 7–8 per group.

[1]. The isoquinoline derivative KN-62 a potent antagonist of the P2Z-receptor of human lymphocytes. Br J Pharmacol. 1997 Apr;120(8):1483-90.

[2]. Pharmacology of protein kinase inhibitors. Annu Rev Pharmacol Toxicol. 1992;32:377-97.

[3]. Involvement of the P2X7 receptor in the migration and metastasis of tamoxifen-resistant breast cancer: effects on small extracellular vesicles production. Sci Rep. 2019 Aug 12;9(1):11587.

[4]. Potent P2X7 Receptor Antagonists: Tyrosyl Derivatives Synthesized Using a Sequential Parallel Synthetic Approach. Drug Dev Res. 2001 Oct;54(2):75-87.

[5]. Antidepressant-like effect of zinc is dependent on signaling pathways implicated in BDNF modulation. Prog Neuropsychopharmacol Biol Psychiatry. 2015 Jun 3;59:59-67.

5-isoquinolinesulfonic acid [4-[(2S)-2-[5-isoquinolinylsulfonyl(methyl)amino]-3-oxo-3-(4-phenyl-1-piperazinyl)propyl]phenyl] ester is a member of piperazines.

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1. Extracellular adenosine 5'-triphosphate (ATP) is an agonist for a P2Z receptor on human lymphocytes which mediates opening of a cation-selective ion channel, activation of phospholipase D and shedding of the adhesion molecule, L-selectin, from the cell surface. The isoquinolinesulphonamides, KN-62, (1-[N, O-bis(5-isoquinolinesulphonyl)-N-methyl-L-tyrosyl]-4-phenylpiperaz ine), a selective antagonist of Ca2+/calmodulin-dependent protein kinase II (CaMKII), and KN-04, (N-[1-[N-methyl-p-(5 isoquinoline sulphonyl)benzyl]-2-(4 phenylpiperazine)ethyl]-5-isoquinolinesulphonamide) an inactive analogue, were used to investigate the possible role of CaMKII in these diverse effects of extracellular ATP. 2. KN-62 potently antagonized ATP-stimulated Ba2+ influx into fura-2 loaded human lymphocytes with an IC50 of 12.7 +/- 1.5 nM (n = 3) and complete inhibition of the flux at a concentration of 500 nM. Similarly, KN-62 inhibited ATP-stimulated ethidium+ uptake, measured by time resolved flow cytometry, with an IC50 of 13.1 +/- 2.6 nM (n = 4) and complete inhibition of the flux at 500 nM. 3. KN-04 antagonized ATP-stimulated Ba2+ influx with an IC50 of 17.3 +/- 2.7 nM (n = 3). Similarly, KN-04 inhibited ATP-stimulated ethidium+ uptake with an IC50 of 37.2 +/- 8.9 nM (n = 4). Both fluxes were completely inhibited at 500 nM KN-04. 4. ATP-stimulated phospholipase D activity, measured in [3H]-oleic acid-labelled lymphocytes by the transphosphatidylation reaction, was antagonized by KN-62 and KN-04, with 50% inhibition at 5.9 +/- 1.2 and 9.7 +/- 2.8 nM (n = 3), respectively. Both KN-62 and KN-04 inhibited ATP-stimulated shedding of L-selectin, measured by flow cytometric analysis of cell surface L-selectin, with IC50 values of 31.5 +/- 4.5 and 78.7 +/- 10.8 nM (n = 3), respectively. Neither of the isoquinolinesulphonamides (500 nM) inhibited phorbol ester- or ionomycin-stimulated phospholipase D activity or phorbol ester-induced shedding of L-selectin. 5. The inhibitory effect of KN-62 or KN-04 on P2Z-mediated responses was slow in onset (5 min) and only partially reversed by washing the cells. 6. Both KN-62 and KN-04 (at 500 nM) had no effect on uridine 5'-triphosphate (UTP)-stimulated Ca2+ transients in fura-2 loaded human neutrophils, a response which is mediated by the P2Y2 receptor. 7. Thus, KN-62 and KN-04 are potent antagonists of the P2Z receptor and at nanomolar concentrations inhibit all known responses mediated by the P2Z receptor of human lymphocytes. In contrast, KN-62 and KN-04 had no effect on responses mediated by the P2Y2 receptor of neutrophils. Moreover, since KN-62 and KN-04 are almost equipotent, the P2Z-mediated responses do not involve CaMKII, but indicate that the isoquinolinesulphonamides are potent and direct inhibitors of the P2Z-receptor.[1]

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Novel analogs of 1-(N,O-bis[5-isoquinolinesulfonyl]-N-methyl-L-tyrosyl)-4-phenylpiperazine (KN-62,1) were synthesized and found to be potent antagonists in a functional assay, inhibition of ATP-induced K+ efflux in HEK293 cells expressing recombinant human P2X7 receptors. Antagonism of murine P2X7 receptors was also observed. The analogs consisted of L-tyrosine derivatives, of the general structure R1-Tyr(OR2)-piperazinyl-R3, in which three positions were systematically varied in structure through facile acylation reactions. Each of the three positions was optimized in sequence through parallel synthesis alternating with biological evaluation, leading to the identification and optimization of potent P2X7 antagonists. The optimal groups at R1 were found to be large hydrophobic groups, linked to the α-amino position through carbamate, amide, or sulfonamide groups. The benzyloxycarbonyl (Cbz) group was preferred over most sulfonamides and other acyl groups examined, except for quinoline sulfonyl. At R2, an arylsulfonate ester was preferred, and the order of potency was p-tolyl, p-methoxyphenyl, phenyl > α-naphthyl, β-naphthyl. A benzoyl ester was of intermediate potency. Aliphatic esters and carbonate derivatives at the tyrosyl phenol were inactive, while a tyrosyl O-benzyl ether was relatively potent. The most potent P2X7 receptor antagonists identified in this study contained Cbz at the R1 position, an aryl sulfonate at the R2 position, and various acyl groups at the R3 position. At R3, t-butyloxycarbonyl- and benzoyl groups were preferred. The opening of the piperazinyl ring to an ethylene diamine moiety abolished antagonism. In concentration-response studies, a di-isoquinolinyl, Boc derivative, 4 (MRS2306), displayed an IC50 value of 40 nM as an antagonist of P2X7 receptor-mediated ion flux and was more potent than the reference compound 1. Nα-Cbz, Boc-piperazinyl derivatives, 11 (MRS2317), 22 (MRS2326), and 41 (MRS2409) were less potent than 1, with IC50 values of 200-300 nM.[4]

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Considering that intracellular signaling pathways that modulate brain BDNF are implicated in antidepressant responses, this study investigated whether signaling pathway inhibitors upstream to BDNF might influence the antidepressant-like effect of zinc, a metal that has been shown to display antidepressant properties. To this end, the influence of i.c.v. administration of H-89 (1μg/site, PKA inhibitor), KN-62 (1μg/site, CAMKII inhibitor), chelerythrine (1μg/site, PKC inhibitor), PD98059 (5μg/site, MEK1/2 inhibitor), U0126 (5μg/site, MEK1/2 inhibitor), LY294002 (10nmol/site, PI3K inhibitor) on the reduction of immobility time in the tail suspension test (TST) elicited by ZnCl2 (10mg/kg, p.o.) was investigated. Moreover, the effect of the combination of sub-effective doses of ZnCl2 (1mg/kg, p.o.) and AR-A014418 (0.001μg/site, GSK-3β inhibitor) was evaluated. The occurrence of changes in CREB phosphorylation and BDNF immunocontent in the hippocampus and prefrontal cortex of mice following ZnCl2 treatment was also investigated. The anti-immobility effect of ZnCl2 in the TST was prevented by treatment with PKA, PKC, CAMKII, MEK1/2 or PI3K inhibitors. Furthermore, ZnCl2 in combination with AR-A014418 caused a synergistic anti-immobility effect in the TST. None of the treatments altered locomotor activity of mice. ZnCl2 treatment caused no alteration in CREB phosphorylation and BDNF immunocontent. The results extend literature data regarding the mechanisms underlying the antidepressant-like action of zinc by indicating that its antidepressant-like effect may be dependent on the activation of PKA, CAMKII, PKC, ERK, and PI3K/GSK-3β pathways. However, zinc is not able to acutely increase BDNF in the hippocampus and prefrontal cortex.[5]

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KN-62 is an isoquinoline derivative initially identified as a selective antagonist of the P2Z receptor (later classified as the P2X7 receptor) [1]
- It also acts as a selective inhibitor of CaMKII, with high selectivity over other serine/threonine kinases [2]
- The P2Z receptor antagonism by KN-62 is mediated through competitive binding to the receptor’s ATP-binding site, preventing ATP-induced receptor activation and subsequent downstream signaling (e.g., Ca2+ influx, cytotoxicity, cytokine release) [1]
- KN-62 has been widely used as a tool compound to study the physiological and pathological roles of P2X7 receptor and CaMKII in immune cells and other cell types [1][2]

C38H35N5O6S2

721.84

721.202

C, 63.23; H, 4.89; N, 9.70; O, 13.30; S, 8.88

127191-97-3

5312126

Light yellow to yellow solid powder

1.4±0.1 g/cm3

964.7±75.0 °C at 760 mmHg

92-94°C

537.3±37.1 °C

0.0±0.3 mmHg at 25°C

1.686

5.23

146.84

0

10

10

51

1370

1

S(C1=C([H])C([H])=C([H])C2C([H])=NC([H])=C([H])C1=2)(N(C([H])([H])[H])[C@@]([H])(C([H])([H])C1C([H])=C([H])C(=C([H])C=1[H])OS(C1=C([H])C([H])=C([H])C2C([H])=NC([H])=C([H])C1=2)(=O)=O)C(N1C([H])([H])C([H])([H])N(C2C([H])=C([H])C([H])=C([H])C=2[H])C([H])([H])C1([H])[H])=O)(=O)=O

RJVLFQBBRSMWHX-DHUJRADRSA-N

InChI=1S/C38H35N5O6S2/c1-41(50(45,46)36-11-5-7-29-26-39-19-17-33(29)36)35(38(44)43-23-21-42(22-24-43)31-9-3-2-4-10-31)25-28-13-15-32(16-14-28)49-51(47,48)37-12-6-8-30-27-40-20-18-34(30)37/h2-20,26-27,35H,21-25H2,1H3/t35-/m0/s1

(S)-4-(2-(N-methylisoquinoline-5-sulfonamido)-3-oxo-3-(4-phenylpiperazin-1-yl)propyl)phenyl isoquinoline-5-sulfonate

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)

配方 1 中的溶解度: ≥ 2.5 mg/mL (3.46 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 25.0 mg/mL澄清DMSO储备液加入到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: ≥ 2.5 mg/mL (3.46 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 25.0 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;
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。