Literature summary for 2.4.2.4 extracted from

Birck, M.; Clinch, K.; Gainsford, G.; Schramm, V.; Tyler, P.
Syntheses of 5-chlorouracils/thymines with 1-[phosphono(methyl/difluoromethyl)]-1,2-unsaturated-moiety-substituted methyl groups at N(1) and human thymidine phosphorylase inhibitory activity (2009), Helv. Chim. Acta, 92, 823-838.

No PubMed abstract available

Cloned(Commentary)

Cloned (Comment)	Organism
expression of the calmodulin binding protein-tagged enzyme in Escherichia coli strain BL21(DE3) cells transformed with the pCal-n-EK/TP and pGroESL vectors	Homo sapiens

Inhibitors

Inhibitors	Comment	Organism
1-[(R)-3-fluoro-2-(phosphonomethoxy)propyl]thymine	-	Homo sapiens
3-benzoyl-5-chloropyrimidine-2,4(1H,3H)-dione	-	Homo sapiens
5-ethyl-1-[(R)-3-fluoro-2-(phosphonomethoxy)propyl]uracil	-	Homo sapiens
5-ethyl-1-[(R)-3-hydroxy-2-(phosphonomethoxy)propyl]uracil	-	Homo sapiens
5-ethyl-1-[(S)-3-fluoro-2-(phosphonomethoxy)propyl]uracil	-	Homo sapiens
5-methyluridine	-	Homo sapiens
diethyl ((2-[(3,4-dihydro-5-methyl-2,4-dioxopyrimidin-1(2H)-yl)methyl]cyclopent-1-en-1-yl)(difluoro)methyl)phosphonate	-	Homo sapiens
diethyl ((2-[(3,4-dihydro-5-methyl-2,4-dioxopyrimidin-1(2H)-yl)methyl]cyclopent-1-en-1-yl)methyl)phosphonate	-	Homo sapiens
diethyl ((2-[(5-chloro-3,4-dihydro-2,4-dioxopyrimidin-1(2H)-yl)methyl]cyclopent-1-en-1-yl)methyl)phosphonate	-	Homo sapiens
diethyl (2-[(5-chloro-3,4-dihydro-2,4-dioxopyrimidin-1(2H)-yl)methyl]benzyl)phosphonate	-	Homo sapiens
diethyl ([2-(hydroxymethyl)cyclopent-1-en-1-yl]methyl)phosphonate	-	Homo sapiens
diethyl [(Z)-4-(5-chloro-3,4-dihydro-2,4-dioxopyrimidin-1(2H)-yl)but-2-en-1-yl]phosphonate	-	Homo sapiens
diethyl [(Z)-4-hydroxybut-2-en-1-yl]phosphonate	-	Homo sapiens
diethyl [2-(hydroxymethyl)benzyl]phosphonate	-	Homo sapiens
disodium ((2-[(3,4-dihydro-5-methyl-2,4-dioxopyrimidin-1(2H)-yl)methyl]cyclopent-1-en-1-yl)(difluoro)methyl)phosphonate	-	Homo sapiens
disodium ((2-[(3,4-dihydro-5-methyl-2,4-dioxopyrimidin-1(2H)-yl)methyl]cyclopent-1-en-1-yl)methyl)phosphonate	-	Homo sapiens
disodium ((2-[(5-chloro-3,4-dihydro-2,4-dioxopyrimidin-1(2H)-yl)methyl]cyclopent-1-en-1-yl)methyl)phosphonate	-	Homo sapiens
disodium (2-[(5-chloro-3,4-dihydro-2,4-dioxopyrimidin-1(2H)-yl)methyl]benzyl)phosphonate	-	Homo sapiens
disodium [(Z)-4-(5-chloro-3,4-dihydro-2,4-dioxopyrimidin-1(2H)-yl)but-2-en-1-yl]phosphonate	-	Homo sapiens
additional information	synthesis of 5-alkyl, 5-aryl, or 5-fluoro derivatives of 1-[2-(phosphonomethoxy)-ethyl]thymine inhibitors that mimic the interatomic distance between the incoming phosphate and leaving pyrimidine groups at the transition state for the putative SN2 mechanism of thymidine phosphorylase, structures, the inhibitors are not effective compared to (phosphonomethoxy)-alkyl-pyrimidine inhibitors, overview. Switching the N1-linked side chains found in 1-[2-(phosphonomethoxy)-ethyl]thymine, 1-[(R)-3-hydroxy-2-(phosphonomethoxy)propyl]thymine and 1-[(R)-3-fluoro-2-(phosphonomethoxy)propyl]thymine to the N3-position of the nucleobases resulted in significant loss of activity. Replacement of the iminopyrrolidine ring found in 5-ethyl-1-[(R)-3-hydroxy-2-(phosphonomethoxy)propyl]uracil with a 3-methylimidazol-3-ium ring leads to a series of less active 5-halo-6-[(3-methylimidazol-3-ium-1-yl)methyl]-uracil chlorides. On the other hand, replacement with a 2-aminoimidazole ring, leads to a number of 6-[(2-aminoimidazol-1-yl)methyl]-5-chloro-(and 5-bromo)uracil hydrochlorides with similar inhibitory strength compared to 5-ethyl-1-[(R)-3-hydroxy-2-(phosphonomethoxy)propyl]uracil, computer-modeling simulations, overview	Homo sapiens
uridine	-	Homo sapiens

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
additional information	-	additional information	steady-state kinetics	Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
thymidine + phosphate	Homo sapiens	-	thymine + 2-deoxy-alpha-D-ribose 1-phosphate	-	r

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant enzyme from Escherichia coli strain BL21(DE3) by calmodulin affinity chromatography, cleavage of the calmodulin binding protein tag by enterokinase, removal by chromatography on soybean trypsin inhibitor-sepharose	Homo sapiens

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
2'-deoxy-5-nitrouridine + phosphate	-	Homo sapiens	5-nitrouracil + 2-deoxy-alpha-D-ribose 1-phosphate	-	?
thymidine + phosphate	-	Homo sapiens	thymine + 2-deoxy-alpha-D-ribose 1-phosphate	-	r

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
6	-	assay at	Homo sapiens

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism	Structure
0.013	-	uridine	pH 6.0, recombinant enzyme	Homo sapiens
0.017	-	5-methyluridine	pH 6.0, recombinant enzyme	Homo sapiens