Cellular Pharmacology Research Articles

An introduction to a symposium dedicated to the scientific achievements of Roger Nicoll

This special symposium issue contains articles based on talks given at a recent 2 day symposium in honour of Roger A Nicoll, Professor in the Department of Cellular and Molecular Pharmacology at the University of California, San Francisco (UCSF). Roger initially trained in medicine but became interested in neuroscience as a result of reading the work of Sir John Eccles. From 1973 to 1975, he worked with Eccles in Buffalo, New York, and then took a faculty position at UCSF in 1975. Here he initially studied synaptic transmission in the olfactory bulb (Jahr & Nicoll, 1982) and hippocampal formation (Alger & Nicoll, 1982) but soon moved almost exclusively to the hippocampus where his laboratory has been studying the neurons and their synaptic connections for the last 30 years. These studies have examined the properties of both excitatory and inhibitory systems and led to fundamental changes in our understanding of both forms of synaptic transmission. His laboratory has been at the forefront of understanding the cellular and biochemical mechanisms that underpin synaptic plasticity, particularly long-term potentiation. He has trained dozens of scientists, many of whom are now are senior lab heads spread over the world. This symposium was a celebration of Roger's life in science, bringing together people who have been in his laboratory as well as his closest colleagues to discuss their areas within the lab as well as areas of their own current research. It was a wonderful event with Professor Eric Kandel giving the keynote address.

Abstract 919: Design and preclinical pharmacological evaluation of a cleavable succinate ester solubilizing group for isoindolinone MDM2-p53 protein-protein interaction inhibitors

Abstract The potent and selective MDM2-p53 antagonist 4-chloro-3-(4-chlorophenyl)-3-((1-(hydroxymethyl)cyclopropyl)methoxy)-2-(4-nitrobenzyl)isoindolin-1-one 1 displays promising activity in cellular tumour models; however, poor solubility precludes in vivo investigations.(Bioorg. Med. Chem. Letters 2011, 21, 5916-5919) A water-soluble succinate ester of 1 was designed as a pro-drug, based on an understanding of the interaction of the parent compound 1 with MDM2, and the cellular pharmacology and in vivo pharmacokinetics of the ester 2 were defined. Compound 2 was synthesised and evaluated as an antagonist of MDM2- and MDMX-p53 using ELISA assays. Cell growth inhibition induced by 2 and 1 were compared in paired p53 functional and p53 non-functional HCT116, A2780 and SJSA-1 cell lines. Conversion of 2 into 1 was studied in human and mouse plasma in vitro, as was uptake into tumour cells, by HPLC analysis. The pharmacokinetics of 2 was studied in vivo in tumour-bearing mice. Unexpectedly, the succinate ester 2 was 7-fold more potent than the parent compound 1 as an MDM2-p53 binding antagonist in cell-free assays (IC50 values 20 ± 13 nM versus 95 ± 96 nM, respectively) and maintained >100-fold selectivity for MDM2 over MDMX antagonism. In cell growth inhibition assays (GI50) 2 was 2.3-, 6.1- and 8.6-fold more potent in paired p53 functional versus non-functional HCT116, A2780 and SJSA-1 cell lines, respectively. As anticipated, the solubility of 2 (65 µM) was greatly improved over 1 (6.5 µM), and the succinate ester was converted to the parent compound in mouse plasma (>90% in 60 min), although interestingly not in human plasma. Following incubation of tumour cell lines with 2, both the pro-drug and parent compound could be detected. Administration of 2 to A2780 tumour-bearing mice i.v. at 10 mg/kg generated plasma concentrations approaching those required for cell growth inhibition (≥ 3 μM), and oral administration at 100 mg/kg resulted in 2 bio-availability of 7%, and an 1 AUC that was estimated to be 21% of the equivalent i.v. dose of 2. However, a major unidentified metabolite was observed which compromised the in vivo utility of the succinate ester 2. Protein-protein interaction inhibitors as exemplified by MDM2-p53 binding antagonists are a major new class of antitumour agent. However, targeting hydrophobic protein-protein interactions can result in molecules with poor physicochemical properties. A cleavable succinate ester approach can be applied to address this problem without compromising target interaction or cellular activity, as in the current study. Although additional metabolite pathways may limit the utility of this approach, the improved potency gained by addition of a solubilising group provides valuable information for further lead optimisation. (This work was supported by the CR-UK) Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 919. doi:1538-7445.AM2012-919

Myocilin Interacts with Syntrophins and Is Member of Dystrophin-associated Protein Complex

Genetic studies have linked myocilin to open angle glaucoma, but the functions of the protein in the eye and other tissues have remained elusive. The purpose of this investigation was to elucidate myocilin function(s). We identified α1-syntrophin, a component of the dystrophin-associated protein complex (DAPC), as a myocilin-binding candidate. Myocilin interacted with α1-syntrophin via its N-terminal domain and co-immunoprecipitated with α1-syntrophin from C2C12 myotubes and mouse skeletal muscle. Expression of 15-fold higher levels of myocilin in the muscles of transgenic mice led to the elevated association of α1-syntrophin, neuronal nitric-oxide synthase, and α-dystroglycan with DAPC, which increased the binding of laminin to α-dystroglycan and Akt signaling. Phosphorylation of Akt and Forkhead box O-class 3, key regulators of muscle size, was increased more than 3-fold, whereas the expression of muscle-specific RING finger protein-1 and atrogin-1, muscle atrophy markers, was decreased by 79 and 88%, respectively, in the muscles of transgenic mice. Consequently, the average size of muscle fibers of the transgenic mice was increased by 36% relative to controls. We suggest that intracellular myocilin plays a role as a regulator of muscle hypertrophy pathways, acting through the components of DAPC.

Cellular Pharmacology of Protein Kinase Mζ (PKMζ) Contrasts with Its in Vitro Profile: IMPLICATIONS FOR PKMζ AS A MEDIATOR OF MEMORY

A number of recent studies have used pharmacological inhibitors to establish a role for protein kinase Mζ (PKMζ) in synaptic plasticity and memory. These studies use zeta inhibitory peptide (ZIP) and chelerythrine as inhibitors of PKMζ to block long term potentiation and memory; staurosporine is used as a negative control to show that a nonspecific kinase inhibitor does not block long term potentiation and memory. Here, we show that neither ZIP nor chelerythrine inhibits PKMζ in cultured cells or brain slices. In contrast, staurosporine does block PKMζ activity in cells and brain slices by inhibiting its upstream phosphoinositide-dependent kinase-1. These studies demonstrate that the effectiveness of drugs against purified PKMζ may not be indicative of their specificity in the more complex environment of the cell and suggest that PKMζ is unlikely to be the mediator of synaptic plasticity or memory.

Antidiabetic Properties of Berberine: From Cellular Pharmacology to Clinical Effects

Berberine is an alkaloid that is highly concentrated in the roots, rhizomes, and stem bark of various plants. It affects glucose metabolism, increasing insulin secretion, stimulating glycolysis, suppressing adipogenesis, inhibiting mitochondrial function, activating the 5' adenosine monophosphate-activated protein kinase (AMPK) pathway, and increasing glycokinase activity. Berberine also increases glucose transporter-4 (GLUT-4) and glucagon-like peptide-1 (GLP-1) levels. On GLP-1 receptor activation, adenylyl cyclase is activated, and cyclic adenosine monophosphate is generated, leading to activation of second messenger pathways and closure of adenosine triphosphate–dependent potassium channels. Increased intracellular potassium causes depolarization, and calcium influx through the voltage-dependent calcium channels occurs. This intracellular calcium increase stimulates the migration and exocytosis of the insulin granules. In glucose-consuming tissues, such as adipose, or liver or muscle cells, berberine affects both GLUT-4 and retinol-binding protein-4 in favor of glucose uptake into cells; stimulates glycolysis by AMPK activation; and has effects on the peroxisome proliferator-activated receptor γ molecular targets and on the phosphorylation of insulin receptor substrate-1, finally resulting in decreased insulin resistance. Moreover, recent studies suggest that berberine could have a direct action on carbohydrate metabolism in the intestine. The antidiabetic and insulin-sensitizing effect of berberine has also been confirmed in a few relatively small, short-term clinical trials. The tolerability is high for low dosages, with some gastrointestinal complaints appearing to be associated with use of high dosages.

Effect of HIV-1 infection and sex on the cellular pharmacology of the antiretroviral drugs zidovudine and lamivudine.

The cellular pharmacology of zidovudine (ZDV) and lamivudine (3TC) in vivo is not completely understood. This prospective longitudinal study investigated the relationship between HIV-1 serostatus, sex, race, and time on therapy with intracellular and plasma ZDV and 3TC concentrations. Of 20 HIV-seronegative and 23 HIV-seropositive volunteers enrolled, 16 (8 women) and 21 (5 women) completed all 12 study days, respectively. Volunteers began ZDV-3TC therapy (plus a third active drug in HIV-seropositive volunteers), and steady-state concentrations (C(ss)) were determined after days 1, 3, 7, and 12. A repeated-measures mixed model was utilized. HIV-seronegative status was associated with 22% (95% confidence interval [CI], 0%, 50%) and 37% (15%, 67%) higher C(ss) estimates compared to those of HIV-seropositive individuals for intracellular ZDV-TP and 3TC-TP levels, respectively. African-Americans had 36% (8%, 72%) higher ZDV-TP estimates than non-African-Americans. Sex was not associated with ZDV-TP or 3TC-TP (P > 0.19). Women had 36% (4%, 78%) higher plasma ZDV, but the effect was lessened when normalized by lean body weight (5% [-19%, 38%]; P = 0.68). Plasma 3TC was 19% (0%, 41%) higher in HIV-seropositive volunteers and 22% (0%, 48%) higher in African American volunteers, but these effects were not significant when corrected for creatinine clearance (7% [-9%, 20%] and -5% [-26%, 12%] for HIV serostatus and race, respectively; P > 0.35). These results suggest that HIV-seropositive status decreases and African American race elevates the cellular triphosphates of ZDV and 3TC. This information extends knowledge of ZDV and 3TC cellular pharmacology in vivo and provides new leads for future cellular pharmacology studies aimed at optimizing HIV prevention/treatment with these agents.

Molecular and Cellular Pharmacology of the Hypoxia-Activated Prodrug TH-302

TH-302 is a 2-nitroimidazole triggered hypoxia-activated prodrug (HAP) of bromo-isophosphoramide mustard currently undergoing clinical evaluation. Here, we describe broad-spectrum activity, hypoxia-selective activation, and mechanism of action of TH-302. The concentration and time dependence of TH-302 activation was examined as a function of oxygen concentration, with reference to the prototypic HAP tirapazamine, and showed superior oxygen inhibition of cytotoxicity and much improved dose potency relative to tirapazamine. Enhanced TH-302 cytotoxicity under hypoxia was observed across 32 human cancer cell lines. One-electron reductive enzyme dependence was confirmed using cells overexpressing human NADPH:cytochrome P450 oxidoreductase and radiolytic reduction established the single-electron stoichiometry of TH-302 fragmentation (activation). Examining downstream effects of TH-302 activity, we observed hypoxia-dependent induction of γH2AX phosphorylation, DNA cross-linking, and cell-cycle arrest. We used Chinese hamster ovary cell-based DNA repair mutant cell lines and established that lines deficient in homology-dependent repair, but not lines deficient in base excision, nucleotide excision, or nonhomologous end-joining repair, exhibited marked sensitivity to TH-302 under hypoxia. Consistent with this finding, enhanced sensitivity to TH-302 was also observed in lines deficient in BRCA1, BRCA2, and FANCA. Finally, we characterized TH-302 activity in the three-dimensional tumor spheroid and multicellular layer models. TH-302 showed much enhanced potency in H460 spheroids compared with H460 monolayer cells under normoxia. Multicellular layers composed of mixtures of parental HCT116 cells and HCT116 cells engineered to express an oxygen-insensitive bacterial nitroreductase showed that TH-302 exhibits a significant bystander effect.

Targeting DNA Topoisomerase I with Non-Camptothecin Poisons

DNA topoisomerase I is required for DNA relaxation during a variety of cellular functions. The identification of camptothecins as specific enzyme poisons and their clinical efficacy have stimulated extensive efforts to exploit topoisomerase I as a therapeutic target for cancer. However, several limitations of camptothecins, such as low solubility and stability, high toxicity, and the occurrence of resistance, have encouraged the development of non-camptothecin topoisomerase I inhibitors. Different natural and synthetic compounds (e.g., indolocarbazoles, dibenzonaphthyridine and indenoisoquinoline) have been extensively studied as alternatives to camptothecins and have been proved to be promising therapeutic agents. In this review, we comparatively evaluate the preclinical results obtained with the different non-camptothecin poisons proposed thus far as topoisomerase I inhibitors, with special reference to cellular pharmacology, and discuss the perspective for their use in the clinical setting.

Tissue engineered human skin equivalents.

Human skin not only serves as an important barrier against the penetration of exogenous substances into the body, but also provides a potential avenue for the transport of functional active drugs/reagents/ingredients into the skin (topical delivery) and/or the body (transdermal delivery). In the past three decades, research and development in human skin equivalents have advanced in parallel with those in tissue engineering and regenerative medicine. The human skin equivalents are used commercially as clinical skin substitutes and as models for permeation and toxicity screening. Several academic laboratories have developed their own human skin equivalent models and applied these models for studying skin permeation, corrosivity and irritation, compound toxicity, biochemistry, metabolism and cellular pharmacology. Various aspects of the state of the art of human skin equivalents are reviewed and discussed.

“Networks” is Different

“Networks” is Different

ChemInform Abstract: Synthesis, Antiviral Activity, Cytotoxicity and Cellular Pharmacology of L‐3′‐Azido‐2′,3′‐dideoxypurine Nucleosides.

AbstractA rapid, simple, and general method using microwave‐assisted transglycosylation reactions for the synthesis of compounds like (IV) is presented.

Synthesis and antiviral activity of 2′-deoxy-2′-fluoro-2′-C-methyl-7-deazapurine nucleosides, their phosphoramidate prodrugs and 5′-triphosphates

Thirty novel α- and β-d-2'-deoxy-2'-fluoro-2'-C-methyl-7-deazapurine nucleoside analogs were synthesized and evaluated for in vitro antiviral activity. Several α- and β-7-deazapurine nucleoside analogs exhibited modest anti-HCV activity and cytotoxicity. Four synthesized 7-deazapurine nucleoside phosphoramidate prodrugs (18-21) showed no anti-HCV activity, whereas the nucleoside triphosphates (22-24) demonstrated potent inhibitory effects against both wild-type and S282T mutant HCV polymerases. Cellular pharmacology studies in Huh-7 cells revealed that the 5'-triphosphates were not formed at significant levels from either the nucleoside or the phosphoramidate prodrugs, indicating that insufficient phosphorylation was responsible for the lack of anti-HCV activity. Evaluation of anti-HIV-1 activity revealed that an unusual α-form of 7-carbomethoxyvinyl substituted nucleoside (10) had good anti-HIV-1 activity (EC(50)=0.71±0.25 μM; EC(90)=9.5±3.3 μM) with no observed cytotoxicity up to 100 μM in four different cell lines.

Antitumor and Antiangiogenic Activities of BMS-690514, an Inhibitor of Human EGF and VEGF Receptor Kinase Families

The extensive involvement of the HER kinases in epithelial cancer suggests that kinase inhibitors targeting this receptor family have the potential for broad spectrum antitumor activity. BMS-690514 potently inhibits all three HER kinases, and the VEGF receptor kinases. This report summarizes data from biochemical and cellular pharmacology studies, as well as antitumor activity of BMS-690514. The potency and selectivity of BMS-690514 was evaluated by using an extensive array of enzymatic and binding assays, as well as cellular assays that measure proliferation and receptor signaling. Antitumor activity was evaluated by using multiple xenograft models that depend on HER kinase signaling. The antiangiogenic properties of BMS-690514 were assessed in a matrigel plug assay, and effect on tumor blood flow was measured by dynamic contrast-enhanced MRI. BMS-690514 is a potent and selective inhibitor of epidermal growth factor receptor (EGFR), HER2, and HER4, as well as the VEGF receptor kinases. It inhibits proliferation of tumor cells with potency that correlates with inhibition of receptor signaling, and induces apoptosis in lung tumor cells that have an activating mutation in EGFR. Antitumor activity was observed with BMS-690514 at multiple doses that are well tolerated in mice. There was evidence of suppression of tumor angiogenesis and endothelial function by BMS-690514, which may contribute to its efficacy. By combining inhibition of two receptor kinase families, BMS-690524 is a novel targeted agent that disrupts signaling in the tumor and its vasculature.

Synthesis, antiviral activity, cytotoxicity and cellular pharmacology of l-3′-azido-2′,3′-dideoxypurine nucleosides

Microwave-assisted optimized transglycosylation reactions were used to prepare eleven modified l-3′-azido-2′,3′-dideoxypurine nucleosides. These l-nucleoside analogs were evaluated against HIV and hepatitis B virus. The l-3′-azido-2′,3′-dideoxypurines nucleosides were metabolized to nucleoside 5′-triphosphates in primary human lymphocytes, but exhibited weak or no antiviral activity against HIV-1. The nucleosides were also inactive against HBV in HepG2 cells. Pre-steady state kinetic experiments demonstrated that the l-3′-azido-2′,3′-dideoxypurine triphosphates could be incorporated by purified HIV-1 reverse transcriptase, although their catalytic efficiency ( k pol/ K d) of incorporation was low. Interestingly, a phosphoramidate prodrug of l-3′-azido-2′,3′-dideoxyadenosine exhibited anti-HIV-1 activity without significant toxicity.

Differences in the Cellular Response and Signaling Pathways between Cisplatin and Monodentate Organometallic Ru(II) Antitumor Complexes Containing a Terphenyl Ligand

The new monofunctional Ru(II)-arene complex [(η⁶-arene)Ru(II)(en)Cl]+, where en = 1,2-diaminoethane and the arene is para-terphenyl (complex 1) exhibits promising cytotoxic effects in human tumor cells including those resistant to conventional cisplatin (J. Med. Chem.2008, 51, 5310). The present study is focused on the cellular pharmacology of 1 to elucidate more deeply the mechanisms underlying its antitumor effects. We have identified several cellular mechanisms induced by 1 in human ovarian carcinoma cells, including inhibition of DNA synthesis, overexpression and activation of p53, expression of proapoptotic proteins p21(WAF1) and Bax, G₀/G₁ arrest, and nuclear fragmentation as a result of apoptotic, and, to a much lower extent, also necrotic processes. Thus, 1 inhibits growth of the cancer cells through induction of apoptotic cell death and G₀/G₁ cell cycle arrest. Further investigations have shown that 1 induces apoptosis by regulating the expression of Bcl-2 family proteins. There were significant differences in cellular responses to the treatment with 1 and with conventional cisplatin, particularly in the kinetics and the extent of these responses. In addition, the distinct p53 activation profile of 1 compared with cisplatin provides an explanation for the activity of this ruthenium drug against cisplatin-resistant cells. Hence complex 1 may provide an alternative therapy in patients with acquired cisplatin resistance, particularly with respect to its very low mutagenicity and different mode of action compared to platinum antitumor drugs in clinical use.

Direct speciation analysis of inorganic elements in single cells using X-ray absorption spectroscopy

Up to now, X-ray absorption spectroscopy (XAS), which includes XANES (X-ray absorption near edge structure) and EXAFS (Extended X-ray absorption fine structure), was mainly used with unfocused beams. The recent development of X-ray focusing optics, with spatial resolution down to the micrometre level while maintaining high photon fluxes, has opened the field of XAS application to single cell analysis. Micro-XAS enables the characterization of the local structure of the elements within subcellular structures in terms of oxidation state, site ligation, and coordination. It has been mainly applied to the determination of trace metal oxidation states using micro-XANES, and in some favorable cases to identify the molecules binding to the metals in cells. Due to the minute quantity of analytes in subcellular compartments, micro-EXAFS is still more challenging to perform than micro-XANES as it requires higher signal to noise ratios. Micro-XAS has proven to be useful in various fields of research such as metal-based neuro-degeneration, cellular pharmacology, trace element physiology and metal toxicology. Micro-XAS presents unique capabilities over other speciation methods because it can be performed in situ, directly in subcellular compartments, without cell fractionation which is prone to modify the chemical element species. However, the stability of chemical element species all along the analytical procedure, from sample preparation to storage and analysis should be tested systematically.

Pharmacological considerations for tenofovir and emtricitabine to prevent HIV infection.

The use of antiretroviral medications in HIV-negative individuals as pre-exposure prophylaxis (PrEP) is a promising approach to prevent HIV infection. Tenofovir disoproxil fumarate (TDF) and emtricitabine exhibit desirable properties for PrEP including: favourable pharmacokinetics that support infrequent dosing; few major drug-drug or drug-food interactions; an excellent clinical safety record; and pre-clinical evidence for efficacy. Several large, randomized, controlled clinical trials are evaluating the safety and efficacy of TDF and emtricitabine for this new indication. A thorough understanding of variability in drug response will help determine future investigations in the field and/or implementation into clinical care. Because tenofovir and emtricitabine are nucleos(t)ide analogues, the HIV prevention and toxicity effects depend on the triphosphate analogue formed intracellularly. This review identifies important cellular pharmacology considerations for tenofovir and emtricitabine, which include drug penetration into relevant tissues and cell types, race/ethnicity/pharmacogenetics, gender, cellular activation state and appropriate episodic or alternative dosing strategies based on pharmacokinetic principles. The current state of knowledge in these areas is summarized and the future utility of intracellular pharmacokinetics/pharmacodynamics for the PrEP field is discussed.

Synthesis and Cellular Pharmacology Studies of a Series of 2-amino-3-aroyl-4-substituted Thiophene Derivatives

Microtubules are dynamic structures that play a crucial role in cellular division and are recognized as an important target for cancer therapy. In search of new compounds with strong antiproliferative activity and simple molecular structure, we have synthesized four different series of compounds in which different substituents were linked to the 4- or 5-position of the 2-amino-3-(3',4',5'-trimethoxybenzoyl) thiophene system. When these compounds were analyzed in vitro for their inhibition of cell proliferation, the 4-aryl substituted derivatives had little activity. In contrast, the presence of a methylene, oxymethyl, aminomethyl or methylenepiperazino moiety between the aryl and the 4-position of the thiophene ring resulted in statistically significant improvement in activity relative to the 4-aryl substituted derivatives. It is noteworthy that the antiproliferative effects of the synthesized compounds were more pronounced against human Molt/4 and CEM as compared with murine L1210 and FM3A cells. The effects of a selected series of compounds on cell cycle progression correlated well with their strong antiproliferative activity and inhibition of tubulin polymerization. We found that the antiproliferative effects of the most active compounds were associated with increase of the proportion of cells in the G2/M and sub-G1 phases of the cell cycle. The structure-activity relationships observed in the series of compounds described here should permit the design of more active molecules.

Copper Transporter 2 Regulates Endocytosis and Controls Tumor Growth and Sensitivity to Cisplatin In Vivo

Copper transporter 2 (CTR2) is one of the four copper transporters in mammalian cells that influence the cellular pharmacology of cisplatin and carboplatin. CTR2 was knocked down using a short hairpin RNA interference. Robust expression of CTR2 was observed in parental tumors grown in vivo, whereas no staining was found in the tumors formed from cells in which CTR2 had been knocked down. Knockdown of CTR2 reduced growth rate by 5.8-fold, increased the frequency of apoptotic cells, and decreased the vascular density, but it did not change copper content. Knockdown of CTR2 increased the tumor accumulation of cis-diamminedichloroplatinum(II) [cisplatin (cDDP)] by 9.1-fold and greatly increased its therapeutic efficacy. Because altered endocytosis has been implicated in cDDP resistance, uptake of dextran was used to quantify the rate of macropinocytosis. Knockdown of CTR2 increased dextran uptake 2.5-fold without reducing exocytosis. Inhibition of macropinocytosis with either amiloride or wortmannin blocked the increase in macropinocytosis mediated by CTR2 knockdown. Stimulation of macropinocytosis by platelet-derived growth factor coordinately increased dextran and cDDP uptake. Knockdown of CTR2 was associated with activation of the Rac1 and cdc42 GTPases that control macropinocytosis but not activation of the phosphoinositide-3 kinase pathway. We conclude that CTR2 is required for optimal tumor growth and that it is an unusually strong regulator of cisplatin accumulation and cytotoxicity. CTR2 regulates the transport of cDDP in part through control of the rate of macropinocytosis via activation of Rac1 and cdc42. Selective knockdown of CTR2 in tumors offers a strategy for enhancing the efficacy of cDDP.

Abstract IA6-1: The evolution of folate analogs: Drugs targeted to folate transporters, drugs targeted to novel enzyme sites, effects on signaling pathways

Abstract IA6-1: The evolution of folate analogs: Drugs targeted to folate transporters, drugs targeted to novel enzyme sites, effects on signaling pathways