Fatih M. Uckun

Metvan: a novel oxovanadium(IV) complex with broad spectrum anticancer activity

Among the 25 bis(cyclopentadienyl)vanadium(IV) and 14 oxovanadium(IV) compounds synthesised and evaluated for anticancer activity, bis(4,7-dimethyl-1,10-phenanthroline) sulfatooxovanadium(IV) (metvan) was identified as the most promising multitargeted anticancer vanadium complex with apoptosis-inducing activity. At nanomolar and low micromolar concentrations, metvan induces apoptosis in human leukaemia cells, multiple myeloma cells and solid tumour cells derived from breast cancer, glioblastoma, ovarian, prostate and testicular cancer patients. It is highly effective against cisplatin-resistant ovarian cancer and testicular cancer cell lines. Metvan is much more effective than the standard chemotherapeutic agents dexamethasone and vincristine in inducing apoptosis in primary leukaemia cells from patients with acute lymphoblastic leukaemia, acute myeloid leukaemia or chronic acute myeloid leukaemia. Metvan-induced apoptosis is associated with a loss of mitochondrial transmembrane potential, the generation of reactive oxygen species and depletion of glutathione. Treatment of leukaemia cells from acute lymphoblastic leukaemia, acute myeloid leukaemia and chronic acute myeloid leukaemia patients with metvan inhibits the constitutive expression as well as the gelatinolytic activities of matrix metalloproteinase-9 and -2. Treatment of human malignant glioblastoma and breast cancer cells with metvan at concentrations > 1 μM is associated with a nearly complete loss of the adhesive, migratory and invasive properties of the treated cancer cell populations. Metvan shows favourable pharmacokinetics in mice and does not cause acute or subacute toxicity at the dose levels tested (12.5 – 50 mg/kg). Therapeutic plasma concentrations ≥ 5 μM, which are highly cytotoxic against human cancer cells, can be rapidly achieved and maintained in mice for at least 24 h after intraperitoneal bolus injection of a single 10 mg/kg non-toxic dose of metvan. Metvan exhibits significant antitumour activity, delays tumour progression and prolongs survival time in severe combined immunodeficient mouse xenograft models of human malignant glioblastoma and breast cancer. The broad spectrum anticancer activity of metvan together with favourable pharmacodynamic features and lack of toxicity warrants further development of this oxovanadium compound as a new anticancer agent. Metvan could represent the first vanadium complex as an alternative to platinum-based chemotherapy.

Targeting Mast Cells in Endometriosis with Janus Kinase 3 Inhibitor, JANEX-1

Endometriosis (EMS) is a chronic inflammatory disease of multifactorial etiology characterized by implantation and growth of endometrial glands and stroma outside the uterine cavity. EMS is a significant public health issue as it affects 15–20% of women in their reproductive age. Clinical symptoms may include pelvic pain, dysmenorrhea, dyspareunia, pelvic/abdominal masses, and infertility. Symptomatic treatments such as surgical resection and/or hormonal suppression of ovarian function and analgesics are not as effective as desired. Consequently, there is an enormous unmet need to develop effective medical therapy capable of preventing the occurrence and recurrence of EMS without undesirable side‐effects. EMS‐associated intra‐abdominal bleeding episodes, local inflammation, adhesions, and i.p. immunologic dysfunction leads to pelvic nociception and pelvic pain. Increasing evidence supports the involvement of allergic‐type inflammation in EMS. Invasion of mast cells, degranulation, and proliferation of interstitial component are observed in endometriotic lesions. Presence of activated and degranulating mast cells within the nerve structures can contribute to the development of pain and hyperalgesia by direct effects on primary nociceptive neurons. Therefore, treatments targeting endometrial mast cells may prove effective in preventing or alleviating EMS‐associated symptoms. The Janus kinase 3 (JAK3) is abundantly expressed in mast cells and is required for the full expression of high‐affinity IgE receptor‐mediated mast cell inflammatory sequelae. JANEX‐1/WHI‐P131 is a rationally designed novel JAK3 inhibitor with potent anti‐inflammatory activity in several cellular and in vivo animal models of inflammation, including mouse models of peritonitis, colitis, cellulitis, sunburn, and airway inflammation with favorable toxicity and pharmacokinetic profile. We hypothesize that JAK3 inhibitors, especially JANEX‐1, may prove useful to prevent or alleviate the symptoms of EMS.

Novel tight binding PETT, HEPT and DABO-based non-nucleoside inhibitors of HIV-1 reverse transcriptase.

Non-nucleoside reverse transcriptase (RT) inhibitors (NNRTIs) are a key component of effective combination antiretroviral therapies for HIV/AIDS. NNRTIs despite their chemical diversity, bind to a common allosteric site of HIV-1 RT, the primary target for anti-AIDS chemotherapy, and noncompetitively inhibit DNA polymerization. NNRTIs currently in clinical use have a low genetic barrier to resistance and therefore, the need for novel NNRTIs active against drug-resistant mutants selected by current therapies is of paramount importance. We describe the chemistry and biological evaluation of highly potent novel phenethylthiazolylthiourea (PETT), 1-[(2-hydroxyethoxy)methyl]-6-(phenylthio)thymine (HEPT) and dihydroalkoxybenzyloxopyrimidine (DABO) derivatives targeting the hydrophobic binding pocket of HIV-1 RT. These NNRTIs were rationally designed by molecular modeling and docking studies using a novel composite binding pocket that predicted how drug-resistant mutations would change the RT binding pocket shape, volume, and chemical make-up and how these changes could affect NNRTI binding. Several ligand derivatization sites were identified for docked NNRTIs that fit the composite binding pocket. The best fit was determined by calculating an inhibition constant (Ludi Ki) of the docked compound for the composite binding pocket. Compounds with a Ludi Ki of < 1 μM were identified as the most promising tight binding NNRTIs. These NNRTIs displayed high selective indices with robust anti-HIV-1 activity against the wild-type and drug-resistant isolates carrying multiple RT gene mutations. The high rate of treatment failure due to the emergence of drug resistance mutations makes the discovery of broad-spectrum PETT, HEPT and DABO-based NNRTIs useful as a component of effective combination regimens.

Targeting Spleen Tyrosine Kinase (SYK) for Treatment of Human Disease

Targeting Spleen Tyrosine Kinase (Syk) for Treatment of Human Disease nSpleen tyrosine kinase (SYK) plays a crucial role in the coordination of immune recognition receptors and orchestrates multiple downstream signaling pathways in various hematopoietic cells. In addition to its well-known function in transducing Fcγ receptor- and B cell receptor-mediated events, SYK signals downstream of a growing list of immunoreceptor pathways that modulate the innate and adaptive responses.

Limitations of the Human-PBL-SCID Mouse Model for Vaginal Transmission of HIV-1

SCID mice reconstituted with human peripheral blood lymphocytes (PBL) are amenable to vaginal transmission of HIV‐1. We investigated the effectiveness of this model to establish systemic HIV‐1 infection.

Substituted heterocyclic thiourea compounds as a new class of anti-allergic agents inhibiting IgE/FcεRI receptor mediated mast cell leukotriene release

Mast cell derived leukotrienes (LTs) play a vital role in pathophysiology of allergy and asthma. We synthesized various analogues of indolyl, naphthyl and phenylethyl substituted halopyridyl, thiazolyl and benzothiazolyl thioureas and examined their in vitro effects on the high affinity IgE receptor/Fc epsilon RI-mediated mast cell leukotriene release. Of the 22 naphthylethyl thiourea compounds tested, there were 7 active compounds and N-[1-(1-naphthyl)ethyl]-N-[2-(ethyl-4-acetylthiazolyl)]thiourea (17 and 16) (IC(50)=0.002 microM) and N-[1-(1R)-naphthylethyl]-N-[2-(5-methylpyridyl)]thiourea (compound 5) (IC(50)=0.005 microM) were identified as the lead compounds. Among the 11 indolylethyl thiourea compounds tested, there were seven active compounds and the halopyridyl compounds N-[2-(3-indolylethyl)]-N-[2-(5-chloropyridyl)]thiourea (24) and N-[2-(3-indolylethyl)]-N-[2-(5-bromopyridyl)]thiourea (25) were the most active agents and inhibited the LTC(4) release with low micromolar IC(50) values of 4.9 and 6.1 microM, respectively. The hydroxylphenyl substituted compounds N-[2-(4-hydroxyphenyl)ethyl]-N-[2-(5-chloropyridyl)]thiourea (37; IC(50)=12.6 microM), N-[2-(4-hydroxyphenyl)ethyl]-N-[2-(5-bromopyridyl)]thiourea (50; IC(50)=16.8 microM) and N-[2-(4-hydroxyphenyl)ethyl]-N-[2-(pyridyl)]thiourea (35; IC(50)=8.5 microM) were the most active pyridyl thiourea agents. Notably, the introduction of electron withdrawing or donating groups had a marked impact on the biological activity of these thiourea derivatives and the Hammett sigma values of their substituents were identified as predictors of their potency. In contrast, experimentally determined partition coefficient values did not correlate with the biological activity of the thiourea compounds which demonstrates that their liphophilicity is not an important factor controlling their mast cell inhibitory effects. These results establish the substituted halopyridyl, indolyl and naphthyl thiourea compounds as a new chemical class of anti-allergic agents inhibiting IgE receptor/Fc epsilon RI-mediated mast cell LTC(4) release. Further lead optimization efforts may provide the basis for new and effective treatment as well as prevention programs for allergic asthma in clinical settings.

Therapeutic innovations against HIV

AIDS has become the leading cause of infection-associated mortality worldwide. Since the 1983 discovery of HIV as the aetiological agent of AIDS, numerous compounds have been developed to prevent or disrupt various stages of HIV replication. Three critical stages in the HIV life cycle have served as the molecular targets for 21 antiviral drugs in clinical use: i) the initial attachment of the virion to immune cells; ii) the transcription of viral RNA to viral DNA; and iii) the processing of gag-pol protein by HIV protease. This patent review discusses the therapeutic innovations against HIV-1, including both patented standard anti-HIV drugs in clinical use as well as promising new anti-HIV drug candidates in the pipeline.

Influence of long-term stability conditions on microbicidal nucleoside prodrug (WHI-07)-loaded gel-microemulsion

The objective of this study was to evaluate the long-term stability of the antiretroviral spermicide WHI-07 (5-bromo-6-methoxy-5,6-dihydro-3′-azidothymidine-5′-(p-bromophenyl)-methoxyalaninyl phosphate) in a polymer-based microemulsion. The recovery and stability of WHI-07 in gelmicroemulsion was examined by a validated high-performance liquid chromatography (HPLC) method. The stability was examined over a period of 24 weeks at 3 controlled temperatures (4°C, 25°C, and 40°C). The recovery of the prodrug from 0.5% to 2.0% WHI-07-loaded gel-microemulsion was 99.8%. HPLC analysis revealed that a 2% WHI-07-loaded gel-microemulsion stored at room temperature and cold temperatures for 24 weeks retained >90% of the prodrug, whereas those stored at 40°C maintained 90% of initial WHI-07 for at least 10 weeks. The observed stability of WHI-07 in gel-microemulsion is of great importance for its widespread utility in various climatological conditions.

Synthesis of a piceatannol analog: Replacement of hydroxy group with amide functionality

Abstract Polyhydroxylated stilbene analogs of piceatannol are shown to possess protein‐tyrosine kinase (PTK) inhibitory activity. We have developed a novel approach to introduce an amide moiety into the structure of piceatannol. The amido substituted stilbene derivative was synthesized in 10 steps with an overall yield of 30%.

Synthesis of Substituted Terphenyl Derivatives

Abstract Several substituted terphenyl derivatives were prepared following two synthetic routes with five steps. Condensation of 4‐amino‐3‐bromo‐benzotrifluoride with 4‐(methylthio) phenyl boronic acid in the presence of sodium carbonate and palladium triphenylphosphine in toluene gave 4‐amino‐3‐(4‐methylthiophenyl)‐benzotrifluoride. Treatment of this compound with osmium tetroxide in the presence of 4‐methyl morpholine‐N‐oxide yielded the corresponding sulfone, which was converted into the iodo derivative using sodium nitrite. The iodo derivative thus obtained was treated first with propylmagnesium chloride and triethylborane and subsequently with hydroxylamine‐O‐sulfonic acid to furnish the target compounds.