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Leupeptin, Microbial (Synonyms: Acetyl-L-leucyl-L-leucyl-L-argininal)

カタログ番号GC10027

可逆性プロテアーゼ阻害剤

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Leupeptin, Microbial 化学構造

Cas No.: 103476-89-7

サイズ 価格 在庫数 個数
10mM (in 1mL DMSO)
$36.00
在庫あり
5mg
$23.00
在庫あり
25mg
$69.00
在庫あり
50mg
$112.00
在庫あり

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Sample solution is provided at 25 µL, 10mM.

Product has been cited by 1 publications

Product Documents

Quality Control & SDS

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Protocol

Leupeptin analysis [1]:

Preparation Method

The concentration of Leupeptin, Microbial was calculated by enzyme inhibition assay using 50 pg of papain and 0.2 mM of N-benzoylL-arginine ethyl ester as the target protease and papain substrate, respectively. Enzyme reactions were carried out at 25 ℃ and pH 6.2.

Applications

Leupeptin, Microbial is a leucine-specific protease and a metalloprotease.

Cell experiment [2]:

Cell lines

MRC-C cell

Preparation Method

To investigate the effect of Leupeptin, Microbial on virus yield, MRC-C cultures in 35 mm dishes were infected by adsorption of virus from 0T ml concentrated suspension of HCV 229E for 1 h at 36 ℃. The cultures were washed twice and overlaid with 2 ml Eagle’s MEM plus 0.2% bovine serum albumin and 20 mm-HEPES buffer pH 7.4. Leupeptin, Microbial at a range of concentrations was added to the virus inoculum, to the maintenance medium and also to maintenance medium used to treat the cultures for 1 h before infection. After 24 h at 36 ℃ in air, the cells were scraped into the medium and disrupted by sonication.

Reaction Conditions

0.4 to l00 µg/ml for 25 h(1 h before infection to 24 h after infection)

Applications

The protease inhibitor Leupeptin, Microbial prevented multiplication of the human coronavirus strain 229E in cultures of MRC-C cells.

Animal experiment [3]:

Animal models

C57BL/6NCrl male mouse

Preparation Method

Mice received i.p. injections of 0.5 ml sterile Phosphate Buffered Saline or 0.5 ml PBS containing 9–40 mg/kg Leupeptin, Microbial hemisulfate.

Dosage form

0, 9, 18 36 and 40 mg/kg Leupeptin, Microbial; Intraperitoneal injection

Applications

Rate of LC3b accumulation after Leupeptin, Microbial treatment was greatest in the liver and lowest in spleen.

References:

[1]. Kim IS, Kim YB, et,al. Characterization of the leupeptin-inactivating enzyme from Streptomyces exfoliatus SMF13 which produces leupeptin. Biochem J. 1998 Apr 15;331 (Pt 2)(Pt 2):539-45. doi: 10.1042/bj3310539. PMID: 9531495; PMCID: PMC1219386.

[2]. Appleyard G, Tisdale M. Inhibition of the growth of human coronavirus 229E by leupeptin. J Gen Virol. 1985 Feb;66 (Pt 2):363-6. doi: 10.1099/0022-1317-66-2-363. PMID: 3968542.

[3]. Haspel J, Shaik RS, et,al. Characterization of macroautophagic flux in vivo using a leupeptin-based assay. Autophagy. 2011 Jun;7(6):629-42. doi: 10.4161/auto.7.6.15100. Epub 2011 Jun 1. PMID: 21460622; PMCID: PMC3127049.

Background

Leupeptin、Microbialは広範なセリンとシステインプロテアーゼ阻害剤です[1]。Leupeptin、Microbialは可逆性のプロテアーゼ阻害剤であり、35 nM、3.4 μM、6 nM、および72 nMのKi値を持つ牛トリプシン、ヒトプラスミナーゼ、牛脾臓カテプシンB、および再生人間カルトリプシンに作用します[7]

プロテアーゼ阻害剤であるLeupeptin、Microbialは、MRC-C細胞の培養において人間コロナウイルス株229Eの増殖を抑制しました[3]。トリプシン様プロテアーゼの阻害作用により、Leupeptin、Microbialは基質菌糸体の発達を維持しますが、静止相培養中のLeupeptin、Microbialのタンパク分解によってトリプシン様プロテアーゼが活性化されるため、基質菌糸体が消化されて空気中菌糸体形成が促進されます[2]。またカルペイン阻害剤であるLeupeptin, Microbialは主にネオマイシンから毛細胞を保護しました[5]。 Leupeptinから得られた肝炎B表面抗原(HBsAg)は細胞懸濁液培養物を最大50%回復させました[6]

Leupeptin、Microbialはマウスでよく耐容されました。 Leupeptin、Microbialは、全組織抽出物およびリソソーム濃縮部分(LE部分)においてLC3b-IIを用量依存的に有意に増加させました。 電子顕微鏡(EM)レベルでは、Leupeptin、Microbialは電子密度の高い小胞構造の蓄積を誘発しました[4]。

References:
[1]. Aoyagi T, Miyata S, et,al. Biological activities of leupeptins. J Antibiot (Tokyo). 1969 Nov;22(11):558-68. doi: 10.7164/antibiotics.22.558. PMID: 4243683.
[2]. Kim IS, Kim YB, et,al. Characterization of the leupeptin-inactivating enzyme from Streptomyces exfoliatus SMF13 which produces leupeptin. Biochem J. 1998 Apr 15;331 ( Pt 2)(Pt 2):539-45. doi: 10.1042/bj3310539. PMID: 9531495; PMCID: PMC1219386.
[3]. Appleyard G, Tisdale M. Inhibition of the growth of human coronavirus 229E by leupeptin. J Gen Virol. 1985 Feb;66 ( Pt 2):363-6. doi: 10.1099/0022-1317-66-2-363. PMID: 3968542.
[4]. Haspel J, Shaik RS, et,al. Characterization of macroautophagic flux in vivo using a leupeptin-based assay. Autophagy. 2011 Jun;7(6):629-42. doi: 10.4161/auto.7.6.15100. Epub 2011 Jun 1. PMID: 21460622; PMCID: PMC3127049.
[5]. Aoyagi T, Miyata S, et,al. Biological activities of leupeptins. J Antibiot (Tokyo). 1969 Nov;22(11):558-68. doi: 10.7164/antibiotics.22.558. PMID: 4243683.
[6]. Coffin AB, Williamson KL, et,al. Profiling drug-induced cell death pathways in the zebrafish lateral line. Apoptosis. 2013 Apr;18(4):393-408. doi: 10.1007/s10495-013-0816-8. PMID: 23413197; PMCID: PMC3627356.
[7]. Ganapathi TR, Sunil Kumar GB, et,al. Analysis of the limitations of hepatitis B surface antigen expression in soybean cell suspension cultures. Plant Cell Rep. 2007 Sep;26(9):1575-84. doi: 10.1007/s00299-007-0379-7. Epub 2007 May 30. PMID: 17534624.

Chemical Properties

Cas No. 103476-89-7 SDF
同義語 Acetyl-L-leucyl-L-leucyl-L-argininal
Chemical Name 2-acetamido-N-(1-((5-((diaminomethylene)amino)-1-oxopentan-2-yl)amino)-4-methyl-1-oxopentan-2-yl)-4-methylpentanamide
Canonical SMILES O=C(C(C([H])([H])C(C([H])([H])[H])([H])C([H])([H])[H])([H])N([H])C(C([H])([H])[H])=O)N([H])C(C(N([H])C(C([H])=O)([H])C([H])([H])C([H])([H])C([H])([H])/N=C(N([H])[H])/N([H])[H])=O)([H])C([H])([H])C(C([H])([H])[H])([H])C([H])([H])[H]
Formula C20H38N6O4.1/2H2SO4 M.Wt 493.6
溶解度 ≥ 24.7mg/mL in DMSO, ≥ 20mg/mL in Water Storage Store at -20°C
General tips Please select the appropriate solvent to prepare the stock solution according to the solubility of the product in different solvents; once the solution is prepared, please store it in separate packages to avoid product failure caused by repeated freezing and thawing.Storage method and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored at -20°C, please use it within 1 month.
To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time.
Shipping Condition Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request.

Complete Stock Solution Preparation Table

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1 mg 5 mg 10 mg
1 mM 2.0259 mL 10.1297 mL 20.2593 mL
5 mM 0.4052 mL 2.0259 mL 4.0519 mL
10 mM 0.2026 mL 1.013 mL 2.0259 mL
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Average Rating: 5 ★★★★★ (Based on Reviews and 30 reference(s) in Google Scholar.)

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