HIF-1α/hypoxia inducible factor 1 α

短杆菌肽 A 对革兰氏阳性菌株（包括对多种药物耐药的菌株）表现出强大的广谱抗生素活性[1]。短杆菌肽 A 的一个缺点是其高溶血活性[1]。
与莫能菌素 (HY-N4302) 类似，短杆菌肽 A（0.1 nM–10 μM，72 小时）会降低 RCC 细胞系的活力[2]。 VHL 和 HIF-1α 表达均不会显着影响短杆菌肽 A 的细胞敏感性[2]。在肾细胞癌细胞中，短杆菌肽 A（1 和 10 μM，48 或 72 小时）会导致非凋亡细胞死亡[2]。在肾细胞癌细胞中，短杆菌肽 A（0–10 μM，24 小时）会导致代谢功能障碍并消耗细胞能量[2]。
HIF-1α 和 HIF-2α 蛋白表达、HIF 转录活性和靶基因表达均降低通过短杆菌肽 A（0–1 μM，24-72 小时）[3]。

短杆菌肽 A（0.11 mg/kg；瘤内注射；每周两次，持续 14 天）可抑制 RCC 肿瘤异种移植物的生长[2]。
短杆菌肽 A（0.22 mg/kg；腹腔注射；每周 3 次）连续 26 天）阻止表达 VHL 的 RCC 肿瘤异种移植物生长和血管生成[3]。

细胞系：A498、786-O、Caki-1、SN12C、ACHN、UMRC6、UMRC6+VHL、HEK293T+pcDNA3、HEK293T+HA-HIF-1α、HEK293T+HA-HIF-1α-mut
浓度：0.1 nM-10 μM
孵育时间：72 小时
结果：降低针对 A498、786-O、Caki-1、SN12C、ACHN、UMRC6、UMRC6+VHL、HEK293T+pcDNA3、HEK293T+HA-HIF 的活力-1α 和 HEK293T+HA-HIF-1α-mut 细胞，IC50 分别为 0.420、0.430、0.228、0.104、0.783、0.253、0.425、0.057、0.058 和 0.067 μM。

Animal Model: Six to eight weeks old, female Nu/J mice without hair were given a subcutaneous injection of a 1.0 × 10^6 SN12C cell suspension in a 50% reduced growth factor Matrigel solution[2].
Dosage: 0.11 mg/kg body weight
Administration: Intratumoral injection, twice weekly for 14 days
Result: The average tumor mass was reduced by approximately 40% without significant toxicity.

16132269 Mice oral LD50 1 gm/kg CRC Handbook of Antibiotic Compounds, Volume 1-, Berdy, J., Boca Raton, FL, CRC Press, 1980, 4(1)(240), 1980
16132269 Mice intraperitoneal LD50 60 mg/kg CRC Handbook of Antibiotic Compounds, Volume 1-, Berdy, J., Boca Raton, FL, CRC Press, 1980, 4(1)(240), 1980
16132269 Mice intravenous LD50 5 mg/kg CRC Handbook of Antibiotic Compounds, Volume 1-, Berdy, J., Boca Raton, FL, CRC Press, 1980, 4(1)(240), 1980

[1]. Discovery of gramicidin A analogues with altered activities by multidimensional screening of a one-bead-one-compound library. Nat Commun. 2020 Oct 1;11(1):4935.

[2]. Gramicidin A induces metabolic dysfunction and energy depletion leading to cell death in renal cell carcinoma cells. Mol Cancer Ther. 2013 Nov;12(11):2296-307.

[3]. Gramicidin A blocks tumor growth and angiogenesis through inhibition of hypoxia-inducible factor in renal cell carcinoma. Mol Cancer Ther. 2014 Apr;13(4):788-99.

group of peptide antibiotics produced by Bacillus brevis. Gram-C or S is a cyclic ten-amino acid polypeptide, while Gram-A, B, and D are linear. Gram-C is one of the two major components of tyrosinins. Gram-A (1) is a peptide antibiotic that disrupts transmembrane ion concentration gradients by forming ion channels in the lipid bilayer. Although it has been used clinically for many years, its application has been limited by its strong hemolytic activity and cytotoxicity to mammals (possibly due to their shared ion transport mechanism). This paper reports an integrated high-throughput strategy for discovering analogs of Compound 1 with different bioactivity profiles. We designed 4096 analog structures to maintain the charge neutrality, hydrophobicity, and channel-forming properties of Compound 1. Through analog synthesis, tandem mass spectrometry sequencing, and three micro-screenings, we finally identified 10 representative analogs. Resynthesis and detailed functional evaluation revealed that all 10 analogs had similar ion channel functions, but differed in cytotoxicity, hemolytic activity, and antibacterial activity. Our large-scale structure-activity relationship study suggests that it is feasible to develop compound 1 analogs that can selectively induce toxicity in target organisms. [1]

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Ion carriers are lipid-soluble organic molecules that disrupt the transmembrane potential of cells by making biological membranes permeable to specific ions. They include mobile carriers that complex with metal cations and channel-forming agents that insert into the membrane to form hydrophilic pores. Although mobile carriers have anticancer properties, research on channel-forming agents is limited. Here, we used the channel-forming ion carrier gramin A to investigate its effects on the growth and survival of renal cell carcinoma (RCC) cells. RCC is a highly histologically heterogeneous malignant tumor that is highly resistant to conventional therapies. We found that gramin A reduced the in vitro viability of various renal cell carcinoma (RCC) cell lines at sub-micromolar concentrations (all IC50 < 1.0 μmol/L). Gram-A toxicity to RCC cells is independent of histological subtype, expression of von Hippel-Lindau tumor suppressor gene and its downstream target gene hypoxia-inducible factor-1α. Gram-A’s effect on reducing cell viability is comparable to or stronger than that of the carrier monensin, depending on the cell line. Mechanistic studies have shown that Gram-A blocks ATP production by inhibiting oxidative phosphorylation and glycolysis, leading to cell energy depletion and non-apoptotic cell death. In addition, Gram-A can also effectively inhibit the growth of in vivo RCC xenograft tumors. These results reveal the new application prospects of Gram-A as a potential therapeutic agent for RCC. [2]

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Ion carriers are hydrophobic organic molecules that disrupt the transmembrane potential of cells by increasing the permeability of the cell membrane to specific ions. Gram-A is a channel-forming ion carrier that can form hydrophilic membrane pores that allow monovalent cations to pass through rapidly. Previously, we found that Gram-A can induce cell energy stress and cell death in renal cell carcinoma (RCC) cell lines. Renal cell carcinoma (RCC) is a refractory cancer characterized by constitutive activation of the transcription factor hypoxia-inducible factor (HIF). In this study, we demonstrated that grammidine A can inhibit HIF in RCC cells. We found that under both normoxic and hypoxic conditions, grammidine A destabilized HIF-1α and HIF-2α proteins, thereby reducing HIF transcriptional activity and the expression of various hypoxia-responsive genes. Mechanistic studies revealed that grammidine A accelerates O₂-dependent HIF downregulation by upregulating the expression of von Hippel-Lindau (VHL) tumor suppressor protein, and VHL targets hydroxylated HIF for proteasomal degradation. Furthermore, grammidine A inhibited the growth of human renal cell carcinoma xenografts without significant toxicity in mice. Tumors treated with grammidine A also exhibited physiological and molecular characteristics consistent with HIF-dependent angiogenesis inhibition. In summary, these results indicate that Gram-A, as a potent HIF inhibitor, plays a novel role in inhibiting tumor growth and angiogenesis in renal cell carcinoma expressing VHL. [3]

C99H140N20O17

1882.29000

1881.07

C, 63.17; H, 7.50; N, 14.88; O, 14.45

11029-61-1

Gramicidin A TFA

16132269

White to off-white solid powder

11.262

548.99

21

17

52

136

3980

14

OCCNC([C@@H](NC(CNC([C@@H](NC(CNC([C@@H](NC(CNC([C@@H](NC([C@H](NC([C@@H](NC)C(C)C)=O)C(C)C)=O)CC1C2=CC=CC=C2NC=1)=O)=O)CC1C2=CC=CC=C2NC=1)=O)=O)CC1C2=CC=CC=C2NC=1)=O)=O)CC1C2=CC=CC=C2NC=1)=O

ZWCXYZRRTRDGQE-LUPIJMBPSA-N

InChI=1S/C99H140N20O17/c1-51(2)37-73(109-86(123)59(17)107-81(122)49-105-96(133)82(55(9)10)106-50-121)89(126)108-60(18)87(124)117-84(57(13)14)98(135)119-85(58(15)16)99(136)118-83(56(11)12)97(134)116-80(44-64-48-104-72-34-26-22-30-68(64)72)95(132)112-76(40-54(7)8)92(129)115-79(43-63-47-103-71-33-25-21-29-67(63)71)94(131)111-75(39-53(5)6)91(128)114-78(42-62-46-102-70-32-24-20-28-66(62)70)93(130)110-74(38-52(3)4)90(127)113-77(88(125)100-35-36-120)41-61-45-101-69-31-23-19-27-65(61)69/h19-34,45-48,50-60,73-80,82-85,101-104,120H,35-44,49H2,1-18H3,(H,100,125)(H,105,133)(H,106,121)(H,107,122)(H,108,126)(H,109,123)(H,110,130)(H,111,131)(H,112,132)(H,113,127)(H,114,128)(H,115,129)(H,116,134)(H,117,124)(H,118,136)(H,119,135)/t59-,60-,73+,74+,75+,76+,77-,78-,79-,80-,82-,83+,84+,85-/m0/s1

(2R)-2-[[(2S)-2-[[2-[[(2S)-2-formamido-3-methylbutanoyl]amino]acetyl]amino]propanoyl]amino]-N-[(2S)-1-[[(2R)-1-[[(2S)-1-[[(2R)-1-[[(2S)-1-[[(2R)-1-[[(2S)-1-[[(2R)-1-[[(2S)-1-[[(2R)-1-[[(2S)-1-(2-hydroxyethylamino)-3-(1H-indol-3-yl)-1-oxopropan-2-yl]amino]-4-methyl-1-oxopentan-2-yl]amino]-3-(1H-indol-3-yl)-1-oxopropan-2-yl]amino]-4-methyl-1-oxopentan-2-yl]amino]-3-(1H-indol-3-yl)-1-oxopropan-2-yl]amino]-4-methyl-1-oxopentan-2-yl]amino]-3-(1H-indol-3-yl)-1-oxopropan-2-yl]amino]-3-methyl-1-oxobutan-2-yl]amino]-3-methyl-1-oxobutan-2-yl]amino]-3-methyl-1-oxobutan-2-yl]amino]-1-oxopropan-2-yl]-4-methylpentanamide

Valinegramicidin A; Valyl gramicidin A; Gramicidin A; 11029-61-1; 1-L-Valinegramicidin A; 4419-81-2; Gramicidin A, 1-L-valine-; GNF-Pf-2578; 1-L-Valinegramicidin A;

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

注意： 请将本产品存放在密封且受保护的环境中，避免吸湿/受潮。

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;
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7、 以上所有助溶剂都可在 Invivochem.cn网站购买。