CH3)2 Research Articles

REDISTRIBUTION AND VIRUS INACTIVATION EFFICACY OF A SILICON PHTHALOCYANINE IN RED BLOOD CELL CONCENTRATES AS A FUNCTION OF DELIVERY VEHICLE

The silicon phthalocyanine, HOSiPcOSi(CH3)2(CH2)3N(CH3)2 (Pc 4), is a new photosensitizer that can inactivate lipid-enveloped viruses in red blood cell concentrates (RBCC) upon exposure to red light. Because Pc 4 is insoluble in water, it was delivered either as an emulsion in saline and cremophor EL (CRM) or as a solution in dimethyl sulfoxide (DMSO). In RBCC, Pc 4 added in either vehicle distributed between the plasma and red blood cells (RBC) in a ratio of 4:6, similar to the ratio of these components in RBCC 3:7 (i.e. a hematocrit of 70%). Light exposure did not affect this distribution and caused only marginal degradation of Pc 4 at a light dose that inactivates > 5 log10 vesicular stomatitis virus (VSV). Among human plasma proteins, Pc 4 bound mainly (about 70%) to lipoproteins and to a lesser extent to albumin and lower molecular weight proteins when delivered in DMSO. When delivered in CRM, distribution between lipoproteins and albumin became more even. Among the lipoproteins Pc 4 bound almost exclusively to very low-density lipoproteins (VLDL) when delivered in DMSO and to both VLDL and low-density lipoproteins when added in CRM. The rate of VSV inactivation was independent of the delivery vehicle but there was less RBC damage, as measured by hemolysis during storage, when Pc 4 was added in CRM. These results indicate that using CRM as emulsifier can enhance the specificity of Pc 4-induced photochemical decontamination of RBCC for transfusion.

Selective protection against IgG binding to red cells treated with phthalocyanines and red light for virus inactivation.

Irradiation with red light of red cells (RBCs) containing the photodynamically active phthalocyanine (Pc) dyes is being studied for inactivation of lipid-enveloped viruses. One of the outstanding problems with this treatment is the binding of IgG to RBCs. The effects of oxygen and type I or type II quenchers on this IgG uptake were evaluated. The Pc compounds used were aluminum phthalocyanine tetrasulfonate (AIPcS4), HOSiPcOSi(CH3)2(CH2)3N(CH3)2 (Pc 4); HOSiPcOSi(CH3)2(CH2)3N+(CH3)3I- (Pc 5); and SiPcOSi[(CH3)2(CH2)3N+(CH3)3](2)2I- (Pc 6). RBCs were analyzed by flow cytometry for the presence of IgG. Irradiation with red light for 30 minutes of RBCs containing either 2 microM Pc 4, 2 microM Pc 5, 2 microM Pc 6, or 6.5 microM AIPcS4 resulted in an uptake of IgG. These conditions completely inactivated the lipid-enveloped vesicular stomatitis virus (VSV) (> 5 log10 kill). IgG uptake was reduced when oxygen was depleted. The addition of reduced glutathione (GSH) or mercaptoethanol prevented the binding of IgG with RBCs treated with AIPcS4, Pc 4, Pc 5, and Pc 6. Specific binding of IgG2 but not of C3d was observed upon irradiation of RBCs with Pc 5 and Pc 6 in the absence of GSH. No gross changes were observed in RBC antigen strength after irradiation with the dyes in the presence of GSH. Inactivation of VSV by Pc plus light was not affected by GSH. Sulfhydryl compounds are useful in preventing IgG binding to RBCs following Pc photosensitization. Since virus inactivation proceeds at the same rate in the presence and the absence of sulfhydryl compounds, their addition to treated RBCs should allow crossmatching for transfusion after treatment. The binding of IgG depends to a large extent on the generation of reactive oxygen species.

The use of Zwittergent 3-14 in the purification of recombinant human interferon-beta Ser17 (Betaseron).

A new method for purifying human interferon-beta SER17 from E. coli-derived inclusion bodies has been developed. This procedure eliminates the need for strong denaturants, such as sodium dodecyl sulfate or chaotropes. The procedure makes use of a nondenaturing detergent and a brief incubation at pH 12 to solubilize interferon-beta Ser17 from inclusion bodies. The detergent used was Zwittergent 3-14 (nonionic and pH-insensitive), which is included in the class of sulfobetaines (RN+ (CH3)2(CH2)xSO3-). Zwittergent 3-14 was used in combination with urea to produce a urea/Zwittergent 3-14 washed inclusion body preparation enriched in human interferon-beta Ser17 (Betaseron). Solubilization of inclusion bodies was accomplished by employing a brief (1 minute) shift to pH 12 in the presence of 2.5% Zwittergent 3-14 followed by rapid adjustment to pH 8.0. Solubilization was complete, and the solution could be rapidly adjusted to pH 8 without any observable precipitation of protein. The resultant supernatant could be successfully subjected to a number of chromatographic and analytic procedures, many of which are not compatible with strong anionic detergents, such as SDS. Betaseron was purified from Zwittergent 3-14 solubilized inclusion body lysates using both ion-exchange and size-exclusion chromatography. Purified Betaseron retained bioactivity and could be refolded by simple dialysis against a nonreducing buffer. This method represents a novel procedure for purifying Betaseron from inclusion bodies using a nondenaturing detergent and ion-exchange chromatography.

New phthalocyanines for photodynamic virus inactivation in red blood cell concentrates.

Cationic phthalocyanines with either aluminum or silicon as the central metal were evaluated for their ability to inactivate viruses in red blood cell concentrates (RBCC) photodynamically. In addition, the virucidal potential of a substituted anionic phthalocyanine, aluminum dibenzodisulfophthalocyanine hydroxide (A1N2SB2POH) was evaluated and compared with that of the much studied anionic aluminum tetrasulfophthalocyanine hydroxide (A1PcS4OH). Based on the rate of inactivation of the lipid-enveloped vesicular stomatitis virus (VSV), the virucidal potential of these phthalocyanines was: HOSiPcOSi(CH3)2(CH2)3N+(CH3)3I- (Pc 5) = SiPc[OSi(CH3)2-(CH2)3N+(CH3)3I-]2 (Pc 6) > A1PcOSi(CH3)2(CH2)3N+(CH3)2(CH2)11CH3I- (Pc 21) = A1N2SB2POH = A1PcS4 > HOSiPc[OSi(CH3)2(CH2)3N+(CH3)2(CH2)11CH3I-]2 (Pc 14) > A1PcOSi(CH3)2(CH2)3N+(CH3)3I- (Pc 2). Phthalocyanine ligand 14 and Pc 21 are new phthalocyanines, made by quaternizing known amino analogues. Compared to VSV, the rate of inactivation of Sindbis virus (another model lipid-enveloped virus) was identical when treated in red blood cells (RBC) with Pc 5 and slightly higher when treated with Pc 6 and A1PcS4OH. Treatment of RBCC containing cell-free human immunodeficiency virus (HIV-1) with Pc 5 or A1PcS4OH required 15 min of irradiation to inactivate (> 5 log10 reduction) the virus. The extent of HIV-1 inactivation with A1N2SB2POH was 3.7 log10 after 60 min of red light exposure. The RBC integrity after photosensitization was measured by the ability of the cells to bind to plates coated with poly-L-lysine, (which reflects the retention of the RBC surface negative charges) and hemolysis of the cells over a 7 day storage period.(ABSTRACT TRUNCATED AT 250 WORDS)

New phthalocyanine photosensitizers for photodynamic therapy.

Six new aluminum and silicon phthalocyanines have been synthesized and their photocytotoxicity toward V79 cells has been studied. The compounds that have been prepared are: A1PcOSi(CH3)2(CH2)3N(CH3)2, I; A1Pc-OSi(CH3)2(CH2)3N(CH3)3+I-, II; CH3SiPcOSi(CH3)2(CH2)3N(CH3)2, III; HOSiPcOSi(CH3)2(CH2)3N(CH3)2, IV; HOSiPcOSi(CH3)2(CH2)3N(CH3)3+I-, V; and SiPc[OSi(CH3)2(CH2)3N(CH3)3+I-]2, VI. Relative growth delay values for compounds I-VI and relative cytotoxicity values for compounds I, II, IV, V and VI have been determined. Compounds I and II have been shown to be comparable in photocytotoxicity to what is presumed to be A1PcOH.xH2O, and compound IV has been shown to have greater activity. The classes of compounds to which these six compounds belong appear to have potential for photodynamic therapy.

Protection by the fluoride ion against phthalocyanine-induced photodynamic killing of Chinese hamster cells.

When a dilute F- solution was added to a culture of Chinese hamster cells that had been preincubated with an aluminium phthalocyanine sensitizer derived from AlPcCl, the photosensitivity of the cells was markedly reduced compared to control cells not treated with F-. Under the same treatment conditions, the reduction in [3H]thymidine incorporation into cellular DNA caused by light and this sensitizer and the production of DNA-protein crosslinks caused by light and this sensitizer were also inhibited by F-. In contrast, the killing of Chinese hamster cells, the reduction of thymidine incorporation by the cells, and the production of DNA-protein crosslinks in the cells caused by the combination of light and either Photofrin II or the silicon phthalocyanine HOSiPcOSi(CH3)2(CH2)3-N(CH3)2 were not inhibited by F-. We conclude that the aluminium phthalocyanine sensitizer used is largely or completely AlPc(OH)(H2O), that it is converted to a fluoro complex by F-, and that this compound probably is a less efficient generator of photochemical damage at a critical cellular target(s) than is AlPc(OH)(H2O). The inhibition of thymidine incorporation and DNA-protein crosslink formation indicates that the effects of F- can be expressed at intracellular sites. It is further concluded that the silicon phthalocyanine sensitizer and Photofrin II do not interact significantly with F-.

SYNTHESIS, CHARACTERIZATION AND SPECTRAL BEHAVIOR OF DIORGANOTELLURIUM(IV) SPECIES HAVING A (Te - S) BOND

Abstract (Te—S) bonded compounds of the general formula [R2Te{S2P(OR')2}2| (R=CH3, C6H5, p-MeOC6H4; R'=CH3, C2H5, CH(CH3)2(CH2)2CH3 and CH2CH(CH3)2] have been synthesized by the reaction of R2TeCl2 with the ammonium salt of O,O';-dialkyldithiophosphoric acid in anhydrous toluene at ambient temperature or by the reaction of R2Te(OR')2 (R'=CH3 or C2H5) with corresponding acids. The compounds are monomeric and non-electrolytes in nature. They decompose (TeIV → + Te11 → Te°) slowly in solvents such as CH2Cl2, CHCl3 or CCl4 at ambient temperature. The decomposition products have been characterized. The interaction of [R2Te{S2P(OR')2}2] with phosphine donor bases results in their complete decomposition yielding tellurium metal. The newly synthesized derivatives have been characterized on the basis of analytical and spectral (IR, 1H. 13C, 31P and 125,Te NMR) data. A possible geometry around the central tellurium atom has been proposed.

Effect of charge separation on methyl orange homologue-ionene interaction.

Methyl Orange and Ethyl Orange were used in the study of the effects of charge separation of polyionic amine (ionene) and hydrophobicity of both the dye and ionene on the interaction between the anionic dye and the cationic polymer. The unit structure of x-y ionene was _??_Br-(CH3)2N+(CH2)xN+ (CH3)2(CH2)yBr-_??_. The x-y for seven ionenes used were 3-3, 3-4, 3-6, 3-8, 3-12, 6-6, and 6-12. The amounts of the dye bound to the ionene were determined by means of equilibrium dialysis procedure. One dye molecule was found to occupy one cationic charged site of ionene. The binding occurred due to the positively cooperative electrostatic and hydrophobic interactions. The results were analysed according to the procedure of McGhee and von Hippel. The interaction constants differed widely from one ionene to another ionene. The binding of the dye to the ionene depended strongly on the charge separation and hydrophobicity of both the dye and the ionene. The entropy change of the interaction process suggested strongly that the dye stacked on the polymer.

Micellar properties of zwitterionic phosphobetaine amphiphiles in aqueous solution: Influence of the intercharge distance

Zwitterionic amphiphiles of the general formula H(CH2)y + N(CH3)2(CH2)n PO2C6H 5 − , where the number of intercharge methylenesn is varied, were studied in dilute aqueous solution. Their critical micellar concentrations show a peculiar variation withn, first increasing asn varies from 1 to 4 and then slowly decreasing as methylenes are added up to 10. This behavior is interpreted as being the consequence of two opposite contributions. The first is the classical CMC lowering due to the increase of hydrophobic character with the total number of methylene groups in the surfactant molecule. The second contribution is the increase in the dipole moment of the zwitterionic headgroup withn, leading to stronger dipole-dipole repulsions between headgroups at the micellar surface. Experimental results suggest that the dipole moment does not increase linearly withn because of the polymethylene chain flexibility. This is supported by13C NMR relaxation experiments.

ChemInform Abstract: Crystal and Molecular Structure of Cyclic αα′‐Silaalkadiynes (‐(CH3)2Si‐CC‐Si(CH3)2(CH2)n‐)2, n = 1‐4

ChemInformVolume 18, Issue 2 Physical Organic Chemistry ChemInform Abstract: Crystal and Molecular Structure of Cyclic αα′-Silaalkadiynes (-(CH3)2Si-CC-Si(CH3)2(CH2)n-)2, n = 1-4 G. AA. ELIASSEN, G. AA. ELIASSEN Dep. Chem., Univ. Oslo, Blindern, 0315 Oslo 3, NorwaySearch for more papers by this authorE. KLOSTER-JENSEN, E. KLOSTER-JENSEN Dep. Chem., Univ. Oslo, Blindern, 0315 Oslo 3, NorwaySearch for more papers by this authorC. ROEMMING, C. ROEMMING Dep. Chem., Univ. Oslo, Blindern, 0315 Oslo 3, NorwaySearch for more papers by this author G. AA. ELIASSEN, G. AA. ELIASSEN Dep. Chem., Univ. Oslo, Blindern, 0315 Oslo 3, NorwaySearch for more papers by this authorE. KLOSTER-JENSEN, E. KLOSTER-JENSEN Dep. Chem., Univ. Oslo, Blindern, 0315 Oslo 3, NorwaySearch for more papers by this authorC. ROEMMING, C. ROEMMING Dep. Chem., Univ. Oslo, Blindern, 0315 Oslo 3, NorwaySearch for more papers by this author First published: January 13, 1987 https://doi.org/10.1002/chin.198702067Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat No abstract is available for this article. Volume18, Issue2January 13, 1987 RelatedInformation

Chemical properties of water-soluble porphyrins. 5. Reactions of some manganese (III) porphyrins with the superoxide and other reducing radicals.

Solution properties of three manganese porphyrins, in monomeric form, were investigated. These were the 'picket-fence-like' porphyrin Mn(III)-alpha,alpha,alpha,beta- tetra-ortho(N-methylisonicotinamidophenyl)porphyrin (Mn(III)PFP) and two 'planar unhindered' porphyrins, the Mn(III)TMPyP (tetrakis (4-N-methylpyridyl)porphyrin) and Mn(III)TAP (tetra(4-N,N,N-trimethylanilinium)porphyrin). The porphyrin properties studied were: the absorption spectra in their manganic and manganous forms; acid/base properties of the aquo complexes; the effect of potential axial ligands (up to a concentration of 0.1 mol dm-3) and their one electron reduction potentials. Knowing these properties, the reaction of the Mn(III) porphyrins with the superoxide radical and other reducing radicals were studied using the pulse radiolysis technique. The second-order reaction rate constant of O2- with the Mn(III) porphyrins, which governs the catalytic efficiency of the metalloporphyrins upon the disproportionation of the superoxide radical, was 5.1 X 10(7) to 4.0 X 10(5) dm3 mol-1 s-1, depending on the pH and the nature of the metalloporphyrin. These values are at least one order of magnitude lower than found for Fe(III)TMPyP. One electron reduction of the three Mn(III) porphyrins by eaq-, CO2-, CH2OH and (CH3)2COH had similar second-order rate constants (10(9)-10(10) dm3 mol-1 s-1). That for (CH3)2(CH2)COH was about 10(5) dm3 mol-1 s-1. Reduction in all cases produced the corresponding Mn(II) porphyrin and no intermediate was found. The oxidation reaction of the Mn(II) porphyrins by O2- was approximately two orders of magnitude faster when compared to the reduction of Mn(III) porphyrins with the same radical. Since the reactivities of O2- towards the three manganese (III) compounds follow their reduction potentials, it is suggested that these reactions are governed by an outer-sphere mechanism. This suggestion is corroborated by the finding that water molecules acting as axial ligands, in these aqueous solution systems, are not replaced by another potential ligand when the latter is in the concentration range of 100 mM or less.

Effect of charge density of cationic polyelectrolytes on complex formation with DNA.

AbstractThe interaction between DNA and ionen polymers, ‐[N+(CH3)2(CH2)mN+(CH3)2(CH2)n], with m‐n of 3–3, 6–6, and 6–10 were examined in order to know how the binding behavior of cationic polymers with DNA depends on the charge density of polycation. The ionen polymer has no bulky side chain and the binding forces with DNA would be attributed mainly to electrostatic interaction. When 3–3 ionen polymers were added to DNA solution, precipitable complexes with the ratio of cationic residue to DNA phosphate (+/−) of 1/1 and the free DNA molecules were segregated, while 6–6 and 6–10 ionen polymers formed soluble complexes with DNA molecules up to (+/−) = 0.5. This suggests that 3–3 ionen polymers bind cooperatively with DNA while 6–6 and 6–10 ionen polymers bind noncooperatively. The cooperative binding of 3–3 ionen polymer and the noncooperative binding of 6–6 ionen polymer were also supported by the thermal melting and recooling profiles from the midpoint between first and second meltings. It was concluded that the charge density of DNA phosphate is a critical value determining whether the ionen polymers bind to DNA by a cooperative or by a noncooperative binding, since the distance between successive cationic charges of 3–3 ionen polymer is shorter than that between successive phosphate charges on DNA double helix and those of 6–6 and 6–10 ionen polymers are longer.

Soap‐based detergent formulations: XVIII. Effect of structure variations on surface‐active properties of sulfur containing amphoteric surfactants

AbstractQuaternary ammonium amphoteric surfactants have been found in the past to be excellent lime soap dispersing agents and detergents but exhibit unusual solubility behavior. In search of a relationship between chemical structure and surface‐active properties, compounds having the following general formula were synthesized: ZN+(CH3)2(CH2)nX−, where Z is C12H25‐, C14H29‐, C16H33‐, or C15H31CONHC3H6‐, n is 2, 3, or 4 and X is SO3‐ or OSO3‐. Tertiary amines were converted to sulfobetaines (X=SO3‐) by reaction with (a) butanesultone (n=4), (b) propanesultone (n=3), or (c) sodium 2‐bromoethanesulfonate (n=2). An alternate synthesis for the sulfoethylbetaines (n=2) involved the reaction of the tertiary amines with ethylene bromide, followed by treatment with sodium sulfite. All sulfated quaternary ammonium compounds (X=OSO3‐) were synthesized by treatment of the tertiary amine with the appropriate chloroalcohol, followed by sulfation with chlorosulfonic acid. The sulfated quaternary ammonium amphoterics are stable to acid hydrolysis, and alkaline stability improves with increasing bridge chain length. Sulfoethyl amphoterics are less water soluble than sulfobutyl, which in turn are less soluble than sulfopropyl derivatives of the same alkyl chain length. For the most part, the sulfated amphoterics are insoluble but are solubilized by soap. The lime soap dispersing properties improve as the carbon chain bridge length increases for both the sulfates and sulfonates. Formulations of tallow soap, amphoteric surfactant, and sodium silicates gave good detergency in most cases.

Electronic Conductivity of Poly(N-alkylene, N, N-dimethylammonium chloride)-7, 7, 8, 8-Tetracyanoquinodimethane Salts

7, 7, 8, 8-Tetracyanoquinodimethane (CQ) salts of n, m-ionene N(CH3)2 CH2 N(CH3)2 (CH2), where (n, m) were (2, 5), (3, 3), (4, 4), (5, 5), (6, 5) and (6, 6), were prepared and their specific resistances r and activation energies Ea were measured.When neutral CQ was doped to simple salts, the valuesof r and Ea decreased and CQ complex salts had minimum values of(r=10 2 10 5 ohm, cm) and (Ea=0.077 0.22 eV).Ea could be correlated with the length of polymethylene chain in polymer, when the number of CH2 groups was less than six.The longer the polymethylene chain, the larger Ea of the simple salts and the smaller Ea of the complex salts became. This suggests that the electric conduction mechanism of simple salts is different from that of complex salts.

Topography of Nucleic Acid Helices in Solutions

Abstract A number of reporter molecules of the structure R-(CH2)n-N+(CH3)2(CH2)mN+(CH3)3·2Br−, where R is a chromophore absorbing in the 300–500 mp region, have been synthesized. The effect of DNA and RNA on the absorption, induced circular dichroism, and proton magnetic resonance spectra is reported. A red shift and a hypochromic effect on the absorption spectra of the bound chromophore is observed. In all cases where R is an “unsymmetrical” 4-nitroaniline chromophore, it is found that DNA and RNA induce an opposite CD in the absorption band of the bound reporter molecules. These results together with PMR studies are interpreted in terms of the structure of the nucleic acid systems in solutions.