396726-03-7

Usage

Uses

1-Hydroxy-2-methyl-2-buten-4-yl 4-Diphosphate is an isoprenoid precursor produced by Plasmodium falciparum that affects A. gambiae s.l. blood meal seeking and feeding behaviours as well as susceptibility to infection.

Biochem/physiol Actions

HMB-PP is an intermediate of the non-mevalonate pathway (MEP pathway) of isoprenoid biosynthesis; it was shown to be a potent γδ T cell activator.

Synthetic route

trans-3-hydroxymethyl-2-butenyl chloride (56904-88-2) → (E)-(4-hydroxy-3-methylbut-2-enyl)phosphonophosphoric acid (396726-03-7)

Conditions	Yield
Stage #1: trans-3-hydroxymethyl-2-butenyl chloride With tris(tetra-n-butylammonium) hydrogen pyrophosphate In acetonitrile at 20℃; for 2h; Stage #2: With DOWEX 50WX8-200; ammonium bicarbonate In isopropyl alcohol pH=8.0; Further stages.;	83%

2C-methyl-D-erythritol 2,4-O-cyclodiphosphate (151435-51-7) → (E)-(4-hydroxy-3-methylbut-2-enyl)phosphonophosphoric acid (396726-03-7)

Conditions	Yield
With Escherichia coli; gcpE genes In water at 37℃; for 20h;
With sodium dithionite; paraquat dichloride In water at 37℃; pH=8; Kinetics; Solvent; aq. buffer; Enzymatic reaction;

((E)-4-bromo-2-methylbut-2-en-1-ol) (918823-42-4) → (E)-(4-hydroxy-3-methylbut-2-enyl)phosphonophosphoric acid (396726-03-7)

Conditions	Yield
With tris(tetrabutylammonium) hydrogen pyrophosphate In acetonitrile at 20℃; for 16h;	36.2 mg

(E)-4-chloro-2-methylbut-2-enal (26394-25-2) → (E)-(4-hydroxy-3-methylbut-2-enyl)phosphonophosphoric acid (396726-03-7)

Conditions	Yield
Multi-step reaction with 2 steps 1.1: 80 percent / NaBH4 / tetrahydrofuran / 5 h / -20 °C 2.1: tris(tetra-n-butylammonium) hydrogen pyrophosphate / acetonitrile / 2 h / 20 °C 2.2: 83 percent / Dowex 50WX8-200; NH4HCO3 / propan-2-ol / pH 8.0 View Scheme

2-hydroxy-2-methyl-3-butenal dimethyl acetal (3330-23-2) → (E)-(4-hydroxy-3-methylbut-2-enyl)phosphonophosphoric acid (396726-03-7)

Conditions	Yield
Multi-step reaction with 3 steps 1.1: 164 mg / NaCl; conc. HCl / CuCl2 / benzene / 0 - 50 °C 2.1: 80 percent / NaBH4 / tetrahydrofuran / 5 h / -20 °C 3.1: tris(tetra-n-butylammonium) hydrogen pyrophosphate / acetonitrile / 2 h / 20 °C 3.2: 83 percent / Dowex 50WX8-200; NH4HCO3 / propan-2-ol / pH 8.0 View Scheme

(2R,3R)-4-hydorxy-3-methyl-2,3-epoxybutanyl diphosphate (1196056-86-6) → (E)-(4-hydroxy-3-methylbut-2-enyl)phosphonophosphoric acid (396726-03-7)

Conditions	Yield
With sodium dithionite; paraquat dichloride In water at 37℃; pH=8; Kinetics; Reagent/catalyst; Solvent; aq. buffer; Enzymatic reaction;

(E)-(4-hydroxy-3-methylbut-2-enyl)phosphonophosphoric acid (396726-03-7) → (2E)-2-methylbut-2-ene-1,4-diol (53627-41-1)

Conditions	Yield
With Escherichia coli Type III S alkaline phosphatase In water

(E)-(4-hydroxy-3-methylbut-2-enyl)phosphonophosphoric acid (396726-03-7) → (E)-1,4-diacetoxy-2-methyl-2-butene (59054-99-8)

Conditions	Yield
Multi-step reaction with 2 steps 1: Escherichia coli Type III S alkaline phosphatase / H2O 2: pyridine / 20 °C View Scheme

(E)-(4-hydroxy-3-methylbut-2-enyl)phosphonophosphoric acid (396726-03-7) → dimethylallyl diphosphate (358-72-5) + isopentenyl diphosphate (358-71-4)

Conditions	Yield
With sodium dithionite; IspH; 6,7-dihydro-2,11-dimethyldipyrido<1,2-a:2',1'-c>pyrazinediium dibromide at 37℃; for 0.5h; aq. buffer; Enzymatic reaction;
With E-4-hydroxy-3-methylbut-2-enyl diphosphate reductase from Aquifex aeolicus Mechanism; Enzymatic reaction;
With Clostridium thermocellum VPI 7372 (E)-1-hydroxy-2-methylbut-2-enyl-4-diphosphate reductase fused to ribosomal protein S1; paraquat dichloride; dithionite(2-) Kinetics; Enzymatic reaction;

Upstream product

• 56904-88-2 trans-3-hydroxymethyl-2-butenyl chloride 
• 918823-42-4 ((E)-4-bromo-2-methylbut-2-en-1-ol) 
• 1196056-86-6 (2R,3R)-4-hydorxy-3-methyl-2,3-epoxybutanyl diphosphate 
• 3330-23-2 2-hydroxy-2-methyl-3-butenal dimethyl acetal 
• 26394-25-2 (E)-4-chloro-2-methylbut-2-enal 
• 151435-51-7 2C-methyl-D-erythritol 2,4-O-cyclodiphosphate 

Downstream Products

• 358-72-5 dimethylallyl diphosphate 
• 358-71-4 isopentyl pyrophosphate 
• 59054-99-8 (E)-1,4-diacetoxy-2-methyl-2-butene 

Relevant academic research and scientific papers

Synthesis of (E)-4-hydroxydimethylallyl diphosphate. An intermediate in the methyl erythritol phosphate branch of the isoprenoid pathway

The syntheses of (E)-1-hydroxy-2-methyl-2-buten-4-yl diphosphate ((E)-4-hydroxydimethylallyl diphosphate, HDMAPP), an intermediate in the methyl erythritol phosphate pathway, and (E)-[4-2H]HDMAPP were accomplished in two steps from (E)-4-chloro-2-methyl-2-butenal. The synthetic route is easily adaptable for the facile incorporation of tritium at C-4 of the diphosphate.

Replacing the pyrophosphate group of HMB-PP by a diphosphonate function abrogates its potential to activate human γδ T cells but does not lead to competitive antagonism

The immunological characterization of (E)-4-hydroxy-3-methyl-but-2-enyl pyrophosphate (HMB-PP), and its methylenediphosphonate analogue, HMB-PCP, is described. With an EC50 of 0.1-0.2 nM, HMB-PP is significantly more potent in stimulating human Vγ9/Vδ2 T cells than any other compound described so far. However, replacing the pyrophosphate by a P-CH2-P function abrogates the bioactivity drastically, with HMB-PCP having a EC50 of only 5.3 μM.

IspG converts an epoxide substrate analogue to (E)-4-hydroxy-3-methylbut-2- enyl diphosphate: Implications for IspG catalysis in isoprenoid biosynthesis

(Chemical Equation Presented) IspG is an intriguing enzyme in bacteria, parasite, and plant isoprenoid biosynthesis, and its catalytic mechanism remains elusive. We report here the synthesis of (2R,3R)-4-hydroxy-3-methyl-2,3- epoxybutanyl diphosphate (Epoxy-HMBPP), a proposed intermediate in one of the frequently cited mechanistic models. We have also demonstrated that this epoxide analogue is a catalytically competent IspG substrate. This study represents the first mechanistic study of this important enzyme.

Isoprenoid biosynthesis in Escherichia coli via the methylerythritol phosphate pathway: Enzymatic conversion of methylerythritol cyclodiphosphate into a phosphorylated derivative of (E)-2-methylbut-2-ene-1,4-diol

A crude cell-free system from an Escherichia coli strain overexpressing the cluster containing the three genes yfgA, yfgB, and gcpE converted 2-C-methyl-D-erythritol 2,4-cyclodiphosphate (1) into a phosphorylated derivative of (E)-2-methylbut-2-ene-1,4-diol (6), which most probably represents a novel intermediate in the methylerythritol phosphate pathway for isoprenoid biosynthesis. The free diol 6 was accumulated by phosphatase treatment of the crude enzyme preparation and was identified by comparison with a synthetic reference.