Chan dcm, fu hn, forsch ra, queener sf, rosowsky a. Evaluation of potent inhibitors of dihydrofolate reductase in a. A class of goldi phosphane complexes have been identified as inhibitors of dihydrofolate reductase dhfr from e. Dihydrofolate reductase from a subline of the ll210 lymphoma was purified by affinity chromatography using substituted sepharose 4b to which was coupled methotrexate, a specific, tight binding inhibitor of the enzyme. Plasmodium dihydrofolate reductase is a second enzyme.
Tapale aissms college of pharmacy, kennedy road, near rto office, pune 411 041, maharashtra, india. Dihydrofolate is a target for antibacterial and antiprotozoan drugs. A key finding of the current study which is important in understanding why tmp binds differently to chicken and e. Thus far, all of the clinically useful drugs of this class have been inhibitors of dihydrofolate reductase dhfr, a key enzyme in the synthesis of thymidylate, and therefore, of dna. Pdf structurebased design of dimeric bisbenzimidazole. Pdf synthesis and biological evaluation of a new series. Tapale aissms college of pharmacy, kennedy road, near rto office, pune 411 041. Dihydrofolate reductase an overview sciencedirect topics. Dihydrofolate reductase inhibitors as antibacterial agents. Clinical pharmacology and resistance to dihydrofolate. Structurebased design of dimeric bisbenzimidazole inhibitors. A novel class of dihydrofolate reductase inhibitors mihailo banjanac1, iva tatic1, zrinka ivezic2, sanja tomic3 and jerka dumic4 1cemdd, glaxosmithkline research centre zagreb, prilaz baruna filipovia 29, hr0 zagreb, croatia 2nabriva therapeutics ag, leberstrasse 20, at1112 vienna, austria.
Badhfr is less sensitive to trimethoprim analogs than is dihydrofolate reductase from other species such as escherichia coli, staphylococcus aureus, and streptococcus pneumoniae. Carboxymethylation by iodoacetate of dihydrofolate reductase from the amethopterinresistant. Received 21 september 2016 accepted 23 september 2016 available online xxxx keywords. Malarial dihydrofolate reductase as a paradigm for drug. As part of a search for dihydrofolate reductase dhfr inhibitors combining the high potency of piritrexim ptx with the high antiparasitic vs. While the functional dihydrofolate reductase gene has been mapped to chromosome 5, multiple intronless processed pseudogenes or dihydrofolate. The crystal structures of two human dihydrofolate reductase hdhfr ternary complexes, each with bound nadph cofactor and a lipophilic antifolate inhibitor, have been determined at atomic resolution.
Dihydrofolate reductase dhfr was discovered in the late 1950s by investigators searching for folatedependent enzymes involved in 1carbon metabolism, with its already known application as anticancer and as antibiotics. Dihydrofolate reductase dhfr catalyzes the nadphdependent reduction of dihydrofolate to tetrahydrofolate, and is a crucial enzyme for the synthesis of purines, pyrimidines and some amino acids. Some of the drugs that inhibit this enzyme can, as in the case of methotrexate, be used as anticancer drugs. Specifically, dhfr catalyzes the reduction of folate and 7,8 dihydrofolate dhf to 5,6,7,8tetrahydrofolate thf. Formation of biofilms is a key virulence factor of s. Function dhfr is a key enzyme in folate metabolism as it is involved in 5,10methylene tetra hydro folate thf generation from 7,8 dihydrofolate dhf. Inhibitors of dhfr must have a sufficient specificity for the dhfr of microorganisms and that of cancerous cells for not inducing too many adverse effects in humans. Early dihydrofolate reductase gene amplification events in. Structurebased design of dimeric bisbenzimidazole inhibitors to an emergent trimethoprimresistant type ii dihydrofolate reductase guides the design of monomeric analogues. The type i dihydrofolate reductase, encoded by plasmid r483 and residing within transposon 7 tn7, had a subunit mol. Comprehensive summaries of uppsala dissertations from the faculty of pharmacy 252.
Pharmacology antibiotics dna, rna, folic acid, protein synthesis inhibitors made easy. Novel inhibitors of leishmanial dihydrofolate reductase. In this work, we study the structureactivity relationship of a series of dihydrofolate reductase inhibitors by twodimensional quantitative activitystructure relationship and threedimensional quantitative activitystructure relationship techniques. Approaches to soft drug analogues of dihydrofolate reductase. The latter mechanism appears to be a common mechanism by which animal cells become resistant to methotrexate following exposure to stepwise in creases in drug concentration in vitro.
Reduction of folate by dihydrofolate reductase from. Usa home product directory biochemicals and reagents enzymes, inhibitors, and substrates application index cell signaling enzymes dihydrofolate reductase dhfr enzyme life science home life science products. Early dihydrofolate reductase gene amplification events in cho cells usually occur on the same chromosome arm as the original locus barbara j. This antimicrobial drug class includes sulfonamides, dihydrofolate reductase inhibitors and combinations of these two subclasses. It catalyzes the reduction of 7,8dihydrofolate dhf to 5,6,7,8tetra. Jul 16, 2012 inhibitors of dihydrofolate reductase in leishmania and trypanosomes. The genetic expression of these hybrid plasmids was studied in purified escherichia coli minicells. Pdf dihydrofolate reductase as a therapeutic target researchgate.
Dihydrofolate reductase is a small enzyme that plays a supporting role, but an essential role, in the building of dna and other processes. Biochemicals that inhibit dhfr have many applications in biochemical and physiological research. Many antifolates are known to inhibit dihydrofolate reductase from murine pneumocystis carinii, with 50%. Detailed annotation on the structure, function, physiology, pharmacology and clinical relevance of drug targets.
Pdf the folate antagonists are an important class of therapeutic compounds, as. Dihydrofolate reductase inhibitor an overview sciencedirect topics. Dihydrofolate reductase assay kit catalog number cs0340 storage temperature 20 c technical bulletin product description dihydrofolate reductase dhfr is a ubiquitous enzyme present in all eukaryotic and prokaryotic cells, playing a key role in thymidine synthesis. Inhibitors of dihydrofolate reductase have antibiotic, antimalarial and antineoplastic properties. Design, synthesis, and antifolate activity of new analogues of piritrexim and other diaminopyrimidine dihydrofolate reductase inhibitors with omegacarboxyalkoxy or omegacarboxy1alkynyl substitution in the side chain. The structure of dihydrofolate reductase journal of biological. Dihydrofolate reductase inhibitors as potential drugs, chemischer informationsdienst, 1982, 2wiley online library pdf info.
Lastly, dual compounds, combining dhfr with other folate receptors. The type ii dihydrofolate reductase, specified by plasmid r67, had a subunit mol. Dihydrofolate reductase dhfr inhibitors pharmacorama. The purified enzyme was subjected to disc gel electrophoresis at ph 8. Approaches to soft drug analogues of dihydrofolate reductase inhibitors. Hamlin biomedical sciences division, lawrence livermore national laboratory, livermore, california 94550 usa.
Joining these chemical groups with a flexible linker could. The compounds screened showed weak activity in both the enzyme assays and. Trimethoprim and other nonclassical antifolates an. It catalyzes tetrahydrofolate regeneration by reduction of dihydrofolate, using nadph as a cofactor. This dual drug target aspect was one of the reasons why dihydrofolate reductase became one of the most intensely studied proteins. While the dihydrofolate reductase enzyme is thought to be present in multiple cellular compartments, it is most particularly localized in the cytosol and the nucleoplasm. Plasmodium falciparum has grown increasingly resistant to antifolates for decades. It manages the state of folate, a snaky organic molecule that shuttles carbon atoms to enzymes that need them in their reactions. Dihydrofolate reductase inhibitors are an important class of drugs, as evidenced by their use as antibacterial, antimalarial, antifungal, and anticancer agents. These compounds, together with the thirty experimentally known inhibitors themselves, were docked into a dhfr crystal structure. Flexible diaminodihydrotriazine inhibitors of plasmodium.
Dihydrofolate reductase dihydrofolate reductase inhibitors. Rational drug design, synthesis and biological evaluation of. Rotation of the peptide plane joining i7 and v8 from its position in the folate. Dihydrofolate reductase dhfr plays a critical role in regulating the metabolism of folate. Design, synthesis, and antifolate activity of new analogues of. Insight into inhibitory activity of mycobacterial dihydrofolate reductase inhibitors by insilico molecular modeling approaches.
Merge compounds generated from known rt inhibitors had an aver age molecular weight of 493 9 da, an average logp of 4. This reduces potential side effects of both agents while providing patients a. A number of compounds were obtained and screened against the enzyme and against the intact parasite leishmania donovani and the related organisms trypanosoma brucei and trypanosoma cruzi. Methotrexate is the dhfr inhibitor used most often in a clinical setting as an. Dihydrofolate reductase as a therapeutic target for. Synthesis and biological evaluation of a new series of dihydrofolate reductase inhibitors based on the 42,6diamino5pyrimidinylalkyllglutamic acid structure. Dhfr has a key role in cell growth and proliferation. Dihydrofolate reductase dhfr is a member of the reductase family of enzymes that is ubiquitously expressed in all organisms. Dihydrofolate reductase inhibitors as potential drugs. Dihydrofolate reductase from bacillus anthracis badhfr a validated drug target in the treatment of the infectious disease, anthrax.
Pdf dihydrofolate reductase dhfr was discovered in the late 1950s. Dihydrofolate reductase, or dhfr, is an enzyme that reduces dihydrofolic acid to tetrahydrofolic acid, using nadph as electron donor, which can be converted to the kinds of tetrahydrofolate cofactors used in 1carbon transfer chemistry. This study focuses on structure of dihydrofolate, its interaction with specific amino acids, mechanism of dhfr catalysis. Combining structure features of tmp as template and amide moieties to synthesis. Progress in understanding the biochemical basis of mechanisms responsible for enzyme selectivity and antiproliferative effects has. Moreover, human dihydrofolate reductase is a target for cytostatic and immunosuppressive agents. Dihydrofolate reductase gene amplification and possible. Trimethoprim and other nonclassical antifolates an excellent template for searching modifications of dihydrofolate reductase enzyme inhibitors skip. Inhibition of sporozoite invasion and inhibition of intrahepatic development nat. Dihydrofolate reductase assay kit cs0340 technical bulletin. The inhibitors were designed to avoid steric clash of the pcl group of the inhibitors with the side. Folate inhibitors antagonize the synthesis of folic acid and are used for treating bacterial, fungal and protozoal infections. Iclaprim a new dihydrofolate reductase inhibitor is currently under stage iii clinical trials for complicated skin and skin structure infections. Streptococcus mutans is a major aetiological agent of dental caries.
Resistant parasites have been shown to carry mutant forms of dihydrofolate reductase dhfr, a component of the bifunctional enzyme dihydrofolate reductase thymidylate synthase dhfrts, which has been shown to be the target of pyrimethamine pyr and cycloguanil cyc. Inhibitors of dihydrofolate reductase, 8oxapteridines thesis presented to the graduate council of the north texas state university in partial fulfillment of the requirements for the degree of master of science by shwuching h. Inhibitors of multiple mutants of plasmodium falciparum. Essential proteinprotein interactions between plasmodium. Some of them have little effect on the human enzyme but a strong effect on the dihydrofolate reductases of bacterial and other parasites. The folate antagonists are an important class of therapeutic compounds, as evidenced by their use as antiinfective, antineoplastic, and antiinflammatory drugs.
Kinetic and chemical mechanism of the dihydrofolate reductase. Dihydrofolate reductase dhfr is an enzyme that catalyzes the nadphdependent reduction of dihydrofolate to tetrahydrofolate. Dihydrofolate reductase inhibitors are an important class of drugs. Dihydrofolate reductase, dhfr, whose role is to regenerate folic acid into its reduced form tetrahydrofolate, is necessary for bacteria, plasmodia and normal and cancerous human cells. A dihydrofolate reductase inhibitor dhfr inhibitor is a molecule that inhibits the function of dihydrofolate reductase, and is a type of antifolate since folate is needed by rapidly dividing cells to make thymine, this effect may be used to therapeutic advantage. By combining an antitnf agent with mtx one can lower the dose of both drugs. A total of 3974 ligmergegenerated compounds were derived from the top thirty experimentally verified dhfr binders listed in the bindingdb database as of october 2011. Pdf insight into inhibitory activity of mycobacterial.
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