Chemistry and Biology of Cylindrols: Novel Inhibitors of Ras Farnesyl-Protein Transferase from Cylindrocarpon lucidum

Farnesyl-protein transferase (FPTase) is an enzyme responsible for the farnesylation of Ras protein. Farnesylation is required for cell-transforming activity in several tumor-types, and therefore, inhibition of FPTase activity may be a potential target for anticancer drugs. Our continued search for novel inhibitors led to the isolation of a number of bicyclic resorcinaldehyde cyclohexanone derivatives named here cylindrols A(1) to A(4), cylindrols B and B(1), and a number of known compounds, from Cylindrocarpon lucidum. The compounds were isolated by bioassay-guided separation using Sephadex LH-20, silica gel, and reverse phase HPLC. Structures were elucidated by extensive application of 2D NMR and X-ray crystallography. The determination of absolute stereochemistry was accomplished by CD measurements. Chemical transformations of the most abundant compound resulted in a number of key derivatives which were critical for the evaluation of structure activity relationship. These compounds are members of ascochlorin family and showed a wide range of inhibitory activity (0.7 &mgr;M to >140 &mgr;M) against FPTase. The FPTase activity was noncompetitive with respect to both substrates. Isolation, structures, chemical transformations, and FPTase activity are discussed in detail.     

Facile synthesis of unsymmetrical benzotetrathiafulvalenes via cleavage of the corresponding hexathioorthooxalates

Unsymmetrical hexathioorthooxalates of types ($\text{1}$) and ($\text{2}$) undergo elimination of dialkyl disulfide on heating in an organic solvent; the reaction, which is catalyzed by acid, proceeds without fission of the central C:C bond and provides the first general, high yield synthesis of unsymmetrical benzotetrathiafulvalenes of types ($\text{3}$) and ($\text{4}$).     

Chemistry of the phenoxathiins and isosterically related heterocycles. XXII. 6,7,9-trimethyl-4-azaphenoxathiin: First reported synthesis of an analog of the 4-azaphenoxathiin ring system

The reaction of 2-chloro-3-thiocyanatopyridine with a substituted spiroepoxycyclohexadienone, which served as a masked phenol with reversed polarity, led to the first reported synthesis of an analog of the 4-azaphenoxathiin ring system. Confirmation of the structure was obtained from the assignment of the ¹³C-nmr spectrum.     

Clorobiocin biosynthesis in Streptomyces: identification of the halogenase and generation of structural analogs

Clorobiocin (clo) and novobiocin (nov) are potent inhibitors of bacterial DNA gyrase. The two substances differ in the substitution pattern at C-8' of the aminocoumarin ring, carrying a chlorine atom or a methyl group, respectively. By gene inactivation, clo-hal was identified as the gene of the halogenase responsible for the introduction of the chlorine atom of clorobiocin. Inactivation of cloZ did not affect clorobiocin formation, showing that this ORF is not essential for clorobiocin biosynthesis. Expression of the methyltransferase gene novO in the clo-hal(-) mutant led to the very efficient formation of a hybrid antibiotic containing a methyl group instead of a chlorine atom at C-8'. Comparison of the antibacterial activity of clorobiocin analogs with -Cl, -H, or -CH(3) at C-8' showed that chlorine leads to 8-fold higher activity than hydrogen and to 2-fold higher activity than a methyl group.     

The LOV2 domain of phototropin: a reversible photochromic switch

Light, oxygen, or voltage (LOV) domains constitute a new class of photoreceptor proteins that are sensitive to blue light through a noncovalently bound flavin chromophore. Blue-light absorption by the LOV2 domain initiates a photochemical reaction that results in formation of a long-lived covalent adduct between a cysteine and the flavin cofactor. We have applied ultrafast spectroscopy on the photoaccumulated covalent adduct state of LOV2 and find that, upon absorption of a near-UV photon by the adduct state, the covalent bond between the flavin and the cysteine is broken and the blue-light-sensitive ground state is regained on an ultrafast time scale of 100 ps. We thus demonstrate that the LOV2 domain is a reversible photochromic switch, which can be activated by blue light and deactivated by near-UV light.     

First Synthesis of 3-Mercapto-2(1H)-pyridinone, a Simple Disubstituted Pyridine Useful for Synthesis of the 4-Azaphenoxathiine Ring System and Its Novel Diazaphenoxathiine Analogs: 1,6-Diazaphenoxathiine and 2,6-Diazaphenoxathiine(1)

3-Mercapto-2(1H)-pyridinone (1) can be synthesized in three simple high-yielding steps from readily available 2-tert-butylthiazolo[4,5-b]pyridine (2). Its disodium salt condenses with o-chloronitrobenzene, 2-chloro-3-nitropyridine, and 3-chloro-4-nitropyridine 1-oxide to give respectively 4-azaphenoxathiine (10), 1,6-diazaphenoxathiine (12), and 2,6-diazaphenoxathiine 2-oxide (14) which reduces to 2,6-diazaphenoxathiine (15). The structures of these previously unreported azaphenoxathiine systems were confirmed by assignment of their respective (13)C NMR spectra.     

Chemistry of the phenoxathiins and isosterically related heterocycles. XXIII . 1-azathianthrene: First reported synthesis of a monoazathianthrene and the investigation of the 13C-NMR spectrum using two-dimensional NMR techniques

The reaction of the dianion of 3-mercaptopyridin-2(1H)-thione with 2-chloronitrobenzene in N,N-dimethylformamide leads to the formation of 1-azathianthrene, the first reported mono-aza analog of the thianthrene ring system. A partial assignment of the ¹³C-nmr spectrum of the title compound is reported, the assignment based on chemical shift arguments, spin-lattice (T₁) relaxation times and ¹H-¹³C spin coupling constants. Amplitude modulated two-dimensional Fourier transform (AM2DFT) techniques were employed for the acquisition of the heteronuclear spin-coupling constants.     

The first example of a liquid crystalline side-chain polymer with bent-core mesogenic units: ferroelectric switching and spontaneous achiral symmetry breaking in an achiral polymer

A dimethylsiloxane diluted polysiloxane side chain co-polymer with non-chiral banana-shaped mesogenic units shows an optically isotropic ferroelectric switching polar smectic C phase (SmCPF) consisting of a conglomerate of homogeneously chiral domains with opposite handedness.