7300-59-6

Usage

Chemical Properties

cream coloured to light yellow powder

Uses

Substrate for gamma-glutamyl transpeptidase

InChI:InChI=1/C11H13N3O5/c12-10(15)6-5-9(11(16)17)13-7-1-3-8(4-2-7)14(18)19/h1-4,9,13H,5-6H2,(H2,12,15)(H,16,17)/t9-/m0/s1

Synthetic route

(S)-2-Benzyloxycarbonylamino-4-(4-nitro-phenylcarbamoyl)-butyric acid (113277-32-0) → L-glutamic acid 5-(4-nitroanilide) (7300-59-6)

Conditions	Yield
With hydrogen; palladium on activated charcoal In methanol

(S)-4-[2-(4-Nitro-phenylcarbamoyl)-ethyl]-5-oxo-oxazolidine-3-carboxylic acid benzyl ester (848215-13-4) → L-glutamic acid 5-(4-nitroanilide) (7300-59-6)

Conditions	Yield
Multi-step reaction with 2 steps 1: lithium hydroxide / tetrahydrofuran; H2O / 0.5 h / 5 °C 2: hydrogen / Pd/C / methanol View Scheme

L-glutamic acid (56-86-0) → L-glutamic acid 5-(4-nitroanilide) (7300-59-6)

Conditions	Yield
Multi-step reaction with 6 steps 1: 98 percent / triethylamine / methanol / 3 h 2: 90 percent / toluenesulfonic acid / toluene / 3 h / Heating 3: thionyl chloride / CH2Cl2 / 0.5 h / Heating 4: pyridine / CH2Cl2 / 0.25 h / 20 °C 5: lithium hydroxide / tetrahydrofuran; H2O / 0.5 h / 5 °C 6: hydrogen / Pd/C / methanol View Scheme

N-benzyloxycarbonyl-L-glutamic acid (1155-62-0) → L-glutamic acid 5-(4-nitroanilide) (7300-59-6)

Conditions	Yield
Multi-step reaction with 5 steps 1: 90 percent / toluenesulfonic acid / toluene / 3 h / Heating 2: thionyl chloride / CH2Cl2 / 0.5 h / Heating 3: pyridine / CH2Cl2 / 0.25 h / 20 °C 4: lithium hydroxide / tetrahydrofuran; H2O / 0.5 h / 5 °C 5: hydrogen / Pd/C / methanol View Scheme

(S)-4-<2'-(Chloroformyl)ethyl>-5-oxooxazolidin-3-carbonsaeure-benzylester (58456-26-1) → L-glutamic acid 5-(4-nitroanilide) (7300-59-6)

Conditions	Yield
Multi-step reaction with 3 steps 1: pyridine / CH2Cl2 / 0.25 h / 20 °C 2: lithium hydroxide / tetrahydrofuran; H2O / 0.5 h / 5 °C 3: hydrogen / Pd/C / methanol View Scheme

(S)-3-[3-(benzyloxycarbonyl)-5-oxooxazolidin-4-yl]propanoic acid (23632-67-9) → L-glutamic acid 5-(4-nitroanilide) (7300-59-6)

Conditions	Yield
Multi-step reaction with 4 steps 1: thionyl chloride / CH2Cl2 / 0.5 h / Heating 2: pyridine / CH2Cl2 / 0.25 h / 20 °C 3: lithium hydroxide / tetrahydrofuran; H2O / 0.5 h / 5 °C 4: hydrogen / Pd/C / methanol View Scheme

N-tert-butoxycarbonyl glutamic acid tert-butyl ester (24277-39-2) + 4-nitro-aniline (100-01-6) → L-glutamic acid 5-(4-nitroanilide) (7300-59-6)

Conditions	Yield
Multi-step reaction with 2 steps 1: HATU; N-ethyl-N,N-diisopropylamine / N,N-dimethyl-formamide / 0.5 h / 120 °C / Microwave irradiation 2: hydrogenchloride / 1,4-dioxane / 4 h / 40 °C View Scheme

C20H29N3O7 → L-glutamic acid 5-(4-nitroanilide) (7300-59-6)

Conditions	Yield
With hydrogenchloride In 1,4-dioxane at 40℃; for 4h;

L-glutamic acid 5-(4-nitroanilide) (7300-59-6) → 4-nitro-aniline (100-01-6)

Conditions	Yield
at 37℃; 0.1 M Tris-HCl buffer, γ-glutamyltransferase from mushroom Lentinus edodes; other enzyme; Michaelis constant Km and Vmax determination;
With religiosin B at 37℃; pH=8; Kinetics; aq. buffer; Enzymatic reaction;
With religiosin C at 37℃; pH=8; Kinetics; Concentration; aq. buffer; Enzymatic reaction;

glycylglycine (556-50-3) + L-glutamic acid 5-(4-nitroanilide) (7300-59-6) → L-α-glutamyl=>glycyl=>glycine (6706-20-3)

Conditions	Yield
With HCl buffer; rat kidney γ-glutamyltranspeptidase pH=8.0; Enzyme kinetics;

L-glutamic acid 5-(4-nitroanilide) (7300-59-6) + D-glutamic acid γ-p-nitroanilide (60133-17-7) → (S)-2-((R)-2-Amino-4-carboxy-butyrylamino)-4-(4-nitro-phenylcarbamoyl)-butyric acid

Conditions	Yield
With HCl buffer; rat kidney γ-glutamyltranspeptidase pH=8.0; Enzyme kinetics;

L-glutamic acid 5-(4-nitroanilide) (7300-59-6) → L-glutamic acid (56-86-0) + 4-nitro-aniline (100-01-6)

Conditions	Yield
With marine bivalve lysozyme; MOPS buffer; Tapes japonica at 37℃; pH=7.0; Enzyme kinetics;
With recombinant Pseudomonas nitroreducens IFO12694 γ-glutamyltranspeptidase at 30℃; for 0.0333333h; pH=10.5; aq. buffer; Enzymatic reaction;
With C-terminal hexahistidine-tagged Geobacillus thermodenitrificans NG80-2 γ-glutamyl transpeptidase at 52℃; pH=8; Kinetics; aq. Tris-HCl buffer; Enzymatic reaction;

glycylglycine (556-50-3) + L-glutamic acid 5-(4-nitroanilide) (7300-59-6) → 4-nitro-aniline (100-01-6)

Conditions	Yield
With Tris-HCl buffer at 37℃; pH=8.0; Enzyme kinetics; Further Variations:; Catalysts;

glycylglycine (556-50-3) + L-glutamic acid 5-(4-nitroanilide) (7300-59-6) → gamma-Glutamyl-glycyl-glycine (13640-39-6) + 4-nitro-aniline (100-01-6)

Conditions	Yield
With γ-glutamyltransferase from Bacillus subtilis In aq. phosphate buffer at 22℃; pH=8.5; pH-value; Enzymatic reaction;

Upstream product

• 24277-39-2 N-tert-butoxycarbonyl glutamic acid tert-butyl ester 
• 100-01-6 4-nitro-aniline 
• 23632-67-9 (S)-3-[3-(benzyloxycarbonyl)-5-oxooxazolidin-4-yl]propanoic acid 
• 58456-26-1 (S)-4-<2'-(Chloroformyl)ethyl>-5-oxooxazolidin-3-carbonsaeure-benzylester 
• 1155-62-0 N-benzyloxycarbonyl-L-glutamic acid 
• 848215-13-4 (S)-4-[2-(4-Nitro-phenylcarbamoyl)-ethyl]-5-oxo-oxazolidine-3-carboxylic acid benzyl ester 
• 113277-32-0 (S)-2-Benzyloxycarbonylamino-4-(4-nitro-phenylcarbamoyl)-butyric acid 
• 56-86-0 L-glutamic acid 

Downstream Products

• 13640-39-6 gamma-Glutamyl-glycyl-glycine 
• 100-01-6 4-nitro-aniline 
• 56-86-0 L-glutamic acid 

Relevant academic research and scientific papers

2-Substituted Nγ-glutamylanilides as novel probes of ASCT2 with improved potency

Herein, we report the discovery and structure-activity relationships (SAR) of 2-substituted glutamylanilides as novel probes of the steric environment comprising the amino acid binding domain of alanine-serine-cysteine transporter subtype 2 (ASCT2). Focused library development led to three novel, highly potent ASCT2 inhibitors, with N-(2-(morpholinomethyl)phenyl)-l-glutamine exhibiting the greatest potency in a live-cell glutamine uptake assay. This level of potency represents a three-fold improvement over the most potent, previously reported inhibitor in this series, GPNA. Furthermore, this and other compounds in the series exhibit tractable chemical properties for further development as potential therapeutic leads.

Nγ-Aryl glutamine analogues as probes of the ASCT2 neutral amino acid transporter binding site

Analogues of l-glutamine were designed and synthesized to test a hydrogen-bond hypothesis between ligand and neutral amino acid transporter ASCT2. The key design feature contains a substituted phenyl ring on the amide nitrogen that contains electron withdrawing and electron donating groups that alter the pKa of the amide NH. Through this study a preliminary binding site map has been developed, and a potent commercially available competitive inhibitor of the ASCT2 transporter has been identified.