Sapan K. Jain

An experimental and theoretical study of excited-state dipole moments of some flavones using an efficient solvatochromic method based on the solvent polarity parameter, ETN

The electronic absorption and fluorescence spectra of some biologically active natural flavones have been recorded at room temperature (298 K) in solvents of different polarities. The effects of the solvents upon the spectral properties are discussed. Difference in fluorescence intensity of flavones has been explained on the basis of intersystem crossing and degree of non-planarity calculated theoretically using Austin Model 1 (AM1) method. Excited-state dipole moments have been determined using the solvatochromic method based on the microscopic solvent polarity parameter, E(N)(T). A reasonable agreement has been observed between experimental and AM1 calculated dipole moment changes. Our results are found to be quite reliable in view of the fact that the correlation of the solvatochromic Stokes shifts with microscopic solvent polarity parameter, E(N)(T) is superior to that obtained using bulk solvent polarity functions for all the systems studied here.

Mechanism of biochemical action of substituted 4-methylbenzopyran-2-ones. Part 9: comparison of acetoxy 4-methylcoumarins and other polyphenolic acetates reveal the specificity to acetoxy drug: protein transacetylase for pyran carbonyl group in proximity to the oxygen heteroatom

The evidences for the possible enzymatic transfer of acetyl groups (catalyzed by a transacetylase localized in microsomes) from an acetylated compound (acetoxy-4-methylcoumarins) to enzyme proteins leading to profound modulation of their catalytic activities was cited in our earlier publications in this series. The investigations on the specificity for transacetylase (TA) with respect to the number and positions of acetoxy groups on the benzenoid ring of coumarin molecule revealed that acetoxy groups in proximity to the oxygen heteroatom (at C-7 and C-8 positions) demonstrate a high degree of specificity to TA. These studies were extended to the action of TA on acetates of other polyphenols, such as flavonoids and catechin with a view to establish the importance of pyran carbonyl group for the catalytic activity. The absolute requirement of the carbonyl group in the pyran ring of the substrate for TA to function was established by the observation that TA activity was hardly discernible when catechin pentacetate and 7-acetoxy-3,4-dihydro-2,2-dimethylbenzopyran (both lacking pyran ring carbonyl group) were used as the substrates. Further, the TA activity with flavonoid acetates was remarkably lower than that with acetoxycoumarins, thus suggesting the specificity for pyran carbonyl group in proximity to the oxygen heteroatom. The biochemical properties of flavonoid acetates, such as irreversible activation of NADPH cytochrome C reductase and microsome-catalyzed aflatoxin B(1) binding to DNA in vitro were found to be in tune with their specificity to TA.

Intramolecular excited-state proton-transfer studies on flavones in different environments.

The absorption and fluorescence spectra of some biologically active flavones have been studied as a function of the acidity (pH/H0) of the solution. Dissociation constants have been determined for the ground and first excited singlet states. The results are compared with those obtained from Forster-Weller calculations. The acidity constants obtained by fluorimetric titration method are in complete agreement (in most of the systems) with ground state data indicating a excited state deactivation prior to prototropic equilibration. Compared to umbelliferones, flavones are only weakly fluorescent in alkaline solution. This behaviour is explained by the small energy difference between the singlet excited state and triplet excited state giving rise to more efficient intersystem crossing. Most of the flavones studied here undergo adiabatic photodissociation in the singlet excited state indicating the formation of an exciplex or a phototautomer.

Calreticulin transacetylase (CRTAase): Identification of novel substrates and CRTAase-mediated modification of protein kinase C (PKC) activity in lymphocytes of asthmatic patients by polyphenolic acetates

Earlier reports from our laboratory established the acetyl transferase function of calreticulin (CRT), enabling CRT to transfer acetyl groups from the acetoxy groups of polyphenolic acetates (PAs) to certain receptor proteins. We have in this paper documented the ability of CRT to catalyze the possible transfer of acetyl moiety from 7-acetamido-4-methylcoumarin (7-N-AMC) to the proteins, glutathione S-transferase (GST), and NADPH cytochrome c reductase, leading to the modification of their catalytic activities. 7-Acetoxy-4-methylthiocoumarin (7-AMTC) compared to 7-acetoxy-4-methylcoumarin (7-AMC) when used as a substrate for calreticulin transacetylase (CRTAase) yielded significantly higher catalytic activity. PM3-optimized geometries suggested that the availability of electrons on the sulfur atom of the thiocarbonyl group of the thiocoumarin may render the substrate binding more favorable to the active site of the enzyme as compared to its oxygen analog. Further CRTAase activity was characterized in the human blood lymphocytes. There was no appreciable difference in CRTAase activity of lymphocytes of asthmatic patients as compared to those of normal subjects. The results presented here highlight for the first time the irreversible inhibition of human blood lymphocytes protein kinase C (PKC) by 7,8-diacetoxy-4-methylcoumarin (DAMC) possibly by way of acetylation. The activity of PKC in lymphocytes of asthmatic patients was found to proportionally increase with the severity of the disease. When PA was incubated with lymphocytes of normal patients, PKC was inhibited marginally. On the other hand, lymphocyte PKC of severe asthmatic patients was inhibited drastically. Several PAs inhibited PKC of asthmatic patients in tune with their specificity to CRTAase. DAMC was found to exert maximum inhibitory action on PKC, while 7,8-dihydroxy-4-methylcoumarin (DHMC), the deacetylated product of DAMC, failed to inhibit PKC. These observations clearly describe DAMC as the novel irreversible inhibitor of PKC, and DAMC may be found useful in the control of inflammation and may serve as a potential drug candidate in the therapy of asthma.