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The new water-soluble photosensitizer 5,10,15,20-tetrakis[3,4-bis(carboxymethyleneoxy)phenyl]chlorin (T3,4BCPC) has been prepared, characterized and labeled with 99mTc radionuclide. The radiotracer was evaluated for tissue distribution in Wistar rats. Accumulation of administrated activities in the liver, kidney, bladder and large intestine at 4 h post-injection indicated that the labeled ligand was largely eliminated through the renal and partly through the hepatobiliary system. In vivo biodistribution studies of the labeled compound were carried out in rodent and murine tumor models in comparison with other tumor-seeking radiopharmaceuticals such as 99mTc(V)-dimercaptosuccinic acid (DMSA), 201thallous chloride (TlCl) and 99mTc-citrate using a gamma camera computer system. In N-nitrosomethylurea (NMU)-induced rat mammary tumors, the labeled ligand showed a five-fold tumor to muscle (T/M) ratio compared to 99mTc(V)-DMSA (3-fold) and 201TlCl (3-fold). In the case of C(3)H/J virus-induced spontaneous mammary tumors, the differences were not marked. However, in the transplanted rat C(6)-glioma, the T/M ratio of the labeled compound was appreciably higher (four-fold) than that noted with 99mTc(V)-DMSA (two-fold), 201TlCl (three-fold) and 99mTc-citrate (more than three-fold). These findings suggest that the radiolabeled T3,4BCPC may have potential for the detection of cancer. In order to ascertain the efficacy of the compound for photodynamic therapy applications, a preclinical PDT study was carried out in fibrosarcoma-bearing mice after injecting 5.0 mg/kg body weight of the T3,4BCPC. A laser dose of 20 mW for 60 s resulted in 80% destruction of tumors. These data suggest that this molecule could be useful for PDT of cancer. The labeled agent could also be useful in monitoring the progression/regression of tumors before, during, and after chemotherapy, radiation therapy or PDT., (Copyright 2002 Elsevier Science B.V.)

A new water-soluble cyclam acid porphyrin (CAP), 5,10,15,20-tetrakis [4-[4',8',11'-tris(carboxymethyl)-1'-(1',4',8',11'-tetraazacyclotetradecane)amidomethyleneoxy]phenyl] porphyrin has been synthesised, characterised and labelled with 99mTc. In vivo distribution studies were performed in C6-gliomas and N-nitroso-N-methylurea (NMU) induced mammary tumour bearing rats and scintiimages were obtained at 5 h post-administration of the labelled ligand using gamma camera computer system. Tumour to muscle (T/M) ratios were determined and compared with currently available tumour seeking radiopharmaceuticals such as 99mTc(V)-DMSA, 99mTc-Citrate and 201TlCl. In the case of NMU induced mammary tumour rats the ratios were 6.93, 1.97, 5.30 and 3.29; while in the case of C6-gliomas the ratios were 5.58, 2.18, 3.96 and 3.02 for 99mTc-CAP, 99mTc(V)-DMSA, 99mTc-Citrate and 203TlCl, respectively.

We have synthesized two water soluble dendritic porphyrins, termed DP1 and DP2 and have successfully radiolabeled them with 99mTc. These 99mTc-labeled porphyrins were administered to C6-glioma tumor bearing Wistar rats and scintiimaging and biodistribution studies were carried out. Tumor to muscle ratios of DP1 and DP2 were 8.0 and 9.7, respectively. These molecules may have potential for tumor imaging and diagnosis and may even prove useful as photosensitizers in photodynamic therapy applications.

Technetium-99m labeled cyclam N-2'-methoxyethyl-2-(3'-nitro-1'-triazole) acetamide (cyclam AK 2123) has been synthesized, radiolabeled and characterized as a hypoxic tumor imaging agent. Radiochemical purity was greater than 95%. Marker biodistribution was measured in normal Wistar strain rats at different time intervals after intra venous (i.v.) administration. In vivo distribution and scintigraphic imaging studies were performed after i.v. injection into mammary tumor-bearing rats using a gamma camera and associated computer. Intratumor partial oxygen pressure (pO2) and oxygen saturation measurements were performed to estimate the oxygenation status of the tumors. Tumor to muscle ratio (T/M) of 99mTc-cyclam AK 2123 was 8.5 which was compared with other tumor seeking radiopharmaceuticals, viz. 99mTc-(V) DMSA (3.07), 99mTc-citrate (5.29) and 201T1C1 (3.29). T/M ratios were also evaluated in comparison with radioiodinated iodoazomycin galactopyronoside (125I-IAZG). The ratio obtained was 18 for 99mTc-cyclam AK 2123 and 20 for 125I-IAZG, respectively. The increased concentration of radioactivity in these tumors suggests that this agent could be labelling hypoxic cells and have utility as an imaging agent.

Using mitochondria isolated from Sarcoma 180 ascites tumour in Swiss mice as a model system, we have evaluated the ability of a novel porphyrin, meso-tetrakis[4-(carboxymethyleneoxy)phenyl]porphyrin (H2T4CPP), to induce damage on photosensitization. Oxidative damage to mitochondria, one of the primary and crucial targets of the photodynamic effect, is assessed by measuring products of lipid peroxidation such as thiobarbituric acid reactive substances (TBARS) and lipid hydroperoxides (LOOH), besides the loss of activity of the mitochondrial marker enzyme succinate dehydrogenase (SDH). Analysis of product formation, the effect of deuteration and selective inhibition by scavengers of reactive oxygen species (ROS) show that the damage observed is due mainly to singlet oxygen (1O2) and to a minor extent to hydroxyl radicals (.OH). The 1O2 generation and triplet lifetime of this porphyrin have also been estimated. Fluorescence spectroscopy, used to ascertain the binding of this porphyrin to the mitochondrial proteins, shows a rapid association within 0-2 h and a decline thereafter. Confocal microscopy reveals intracellular localisation of this porphyrin in cells in vitro. Our overall results suggest that the porphyrin H2T4CPP, due to its ability to bind to mitochondrial protein components and to generate ROS upon photoexcitation, may have potential applications in photodynamic therapy.

The ability of a novel porphyrin, meso-tetrakis[3,4-bis(carboxymethyleneoxy)phenyl]porphyrin (T3,4-CPP), to induce photodamage in subcellular membranes, in the form of rat hepatic and tumor microsomes, was evaluated with a view to locating suitable porphyrin derivative for possible use in photodynamic therapy. This water-soluble porphyrin, on exposure to visible light, induced a significant extent of membrane lipid peroxidation as assessed by the formation of thiobarbituric acid reactive substances, lipid hydroperoxides and conjugated dienes. The peroxidation induced in hepatic microsomes is both time- and concentration-dependent. Using inhibitors of reactive oxygen species and comparing products of peroxidation, it is shown that the damage induced is mainly due to singlet oxygen and partly due to other species like free radicals. T3,4-CPP also caused the generation of singlet oxygen as a function of illumination time. Since membrane damage induced by a sensitizer on photoexcitation has been considered to be an important mechanism by which photodynamic cell killing of tumor occurs, the studies on this novel porphyrin indicate the possible potential of this compound in photodynamic therapy.

meso-Tetrakis[4-(carboxymethyleneoxy)phenyl]porphyrin (H2T4CPP) cleaves pBR322 plasmid DNA to single strand breaks in the presence of molecular oxygen and visible light. The above photocleavage was much more efficient in D2O buffer of sodium phosphate (pD = 7.4) than H2O buffer of sodium phosphate (pH = 7.4). In addition this photocleavage of plasmid DNA was inhibited in the presence of sodium azide, lipoic acid, tert-butanol or mannitol suggesting the involvement of 1O2 and.OH in the photocleavage of plasmid DNA. The photocleavage was observed to be more efficient in the presence of H2T4CPP than in the presence of H2CPP [meso-tetrakis (4-carboxy-phenyl)porphyrin]. Our spectral studies using UV-visible, fluorescence and circular dichroism techniques suggest that H2T4CPP binds to DNA while H2CPP does not. Thus, the difference in photocleavage may be caused by the nonbinding of H2CPP and by the binding of H2T4CPP to calf thymus (CT) DNA.

Free radical-induced oxidative damage is involved in several pathological disorders. On the other hand, selective induction of peroxidation in diseased tissue is a promising approach to the treatment of cancer by photodynamic therapy. In this study we have used rat brain mitochondria as a model to evaluate the ability of a new water soluble porphyrin, 5,10,15,20-tetrakis[4-(carboxymethyleneoxy)phenyl]porphyrin (T4CPP), to induce peroxidative damage during photosensitization. Peroxidation in mitochondria, one of the crucial targets of the photodynamic effect, was assessed from the formation of thiobarbituric acid reactive substances and lipid hydroperoxides. The effect on mitochondrial function was estimated from the loss of a mitochondrial marker enzyme, succinate dehydrogenase (SDH). The photodamage was observed to be time- and concentration-dependent of T4CPP. Inhibition studies suggested involvement of singlet oxygen ((1)O2) and, to a lesser extent, of hydroxyl (OH), peroxyl (ROO(-)) and superoxide radicals (O2(-)) in the photodamage. The addition of γ-linolenic acid (a promoter of lipid peroxidation) to the system led to an enhancement of the T4CPP-induced peroxidative damage. Thus, our study indicated that the combination of γ-linolenic acid and T4CPP could enhance the photodynamic effect and has potential applications in photodynamic therapy.

Porphyrins, in combination with light, offer an alternate approach to the treatment of cancer, in the form of photodynamic therapy (PDT). With a view to locate new porphyrins for use in PDT, we evaluated the ability of a novel water-soluble porphyrin, meso-tetrakis[4-(carboxymethyleneoxy)phenyl]porphyrin (T4CPP) to induce photodamage in membranes, using rat hepatic microsomes as a model system. Hepatic microsomes treated with T4CPP and exposed to visible light showed significant lipid peroxidation, as assessed by the formation of conjugated dienes, lipid hydroperoxides, and thiobarbituric acid-reactive substances. The peroxidation induced was both time- and concentration-dependent. T4CPP plus light also resulted in the destruction of the microsomal enzymes adenosine triphosphatase and glucose-6-phosphatase. Analysis of the products of peroxidation and selective inhibition by specific inhibitors showed that the oxidative damage induced was mainly due to singlet oxygen and partly due to hydroxyl radical. The porphyrin T4CPP was efficiently labeled with 99mTc. When this 99mTc-labeled porphyrin was injected into a mammary-tumor-bearing rat, it accumulated in the tumor. Our studies suggest that T4CPP, due to its potential to localize in tumors and to induce membrane damage as exemplified by alteration in rat liver microsomes, may have possible applications in this new modality of cancer treatment.

With a view to locate porphyrins for use in photodynamic therapy (PDT), the new modality of cancer treatment we have evaluated the ability of a novel water soluble porphyrin meso-tetrakis[4-(carboxymethyleneoxy)phenyl]porphyrin (T4CPP) to induce damage to mitochondria during photosensitization. T4CPP, when exposed to visible light, induced lipid peroxidation in rat liver mitochondria as assessed by the formation of thiobarbituric acid reactive substances (TBARS), conjugated dienes (CD) and lipid hydroperoxides (LOOH). The effect on mitochondrial function was assessed by estimating the activity of succinate dehydrogenase (SDH). The peroxidation induced was observed to be time- and concentration- dependent. Analysis of product formation and selective inhibition by scavengers of reactive oxygen species showed that the oxidative damage observed was mainly due to singlet oxygen ((1)O2) and partly due to other reactive species. T4CPP plus light also caused significant lipid peroxidation in Sarcoma 180 ascites tumour mitochondria. Our studies indicate that T4CPP has the potential to photoinduce damage in hepatic and ascites mitochondria, a crucial site of damage in PDT.

Seven new water-soluble cationic complexes of general formula [Pt(2-pyc)(N-N)]+ (where N-N is 2NH3, ethylenediamine (en), 1,2-diaminopropane (1,2-dap), 1,3-diaminopropane (1,3-dap), (+/-) trans-1,2-diaminocyclohaxane (dach), 2,2'-dipyridylamine (dpa) or 1,10-phenanthroline (phen), and 2-pyridinecarboxylate anion) have been prepared. These complexes have been characterized by conductance measurements, and by ultraviolet-visible, infrared (IR), and 1H nuclear magnetic resonance (NMR) spectroscopy. The COSY (correlated spectroscopy) spectra of [Pt(2-pyc)(dpa)]+ and [Pt(2-pys)(dpa)]+ further support the structures of the above complexes with three nitrogen and one oxygen donor atoms in the first coordination sphere of platinum(II) with 1,2-diamine or alpha-diimine and 2-pyridinecarboxylate anion behaving as bidentate ligands. One of the compounds, [Pt(2-pyc)(dpa)]Cl, also shows a birefringence property in water. These compounds inhibit the growth of P388 lymphocytic leukemia cells. [Pt(2-pyc)(dpa)]+ shows I.D.50 value comparable to cisplatin. However, six other complexes show higher I.D.50 values than cisplatin. In addition, the inhibition studies also suggest that their target is DNA. Therefore, the interactions of four of the above complexes with calf thymus DNA have been studied by ultraviolet and fluorescence spectral methods. These studies suggest that [Pt(2-pyc)(NH3)2]+ and [Pt(2-pyc)(1,2-dap)+ bind to DNA by noncovalent interactions. On the other hand, [Pt(2-pyc)(dpa)]+ and [Pt(2-pyc)(phen)]+ bind to DNA by covalent monofunctional binding. The latter two complexes have also been interacted with PUC19 DNA. The gel electrophoresis studies of these interactions suggest that these complexes bind to DNA, and this binding leads to a conformational change in DNA.

Four new palladium(II) and platinum(II) complexes of formula [M(dipy)(AA)]+ (where dipy is 2,2'-dipyridylamine, AA is an anion of glycine or L-alanine, and M is Pd(II) or Pt(II)) have been synthesized and characterized with amino acids binding as bidentate ligands. These complexes are 1:1 electrolyte in conductivity water. Of the above four complexes, the two L-alanine complexes show ID50 values against P388 lymphocytic leukemia cells lower than cis-diamminedichloroplatinum(II), whereas the two glycine complexes show ID50 values higher than cisplatin. The interaction of calf thymus DNA with the above complexes shows significant spectral changes in the presence of [Pt(dipy)(gly)]Cl, [Pd(dipy)(ala)]Cl, and [Pt(dipy)(ala)]Cl and the mode of binding between these complexes and DNA seems to be noncovalent.

Four water soluble complexes of the type [M(bpy)(a-x)]NO3, where M is Pd(II) or Pt(II), bpy is 2,2-bipyridine, and a-x is anion of 2-pyridinecarboxylic acid or 2-pyrazinecarboxylic acid, have been found to bind calf thymus DNA, possibly through hydrogen binding. [M(bpy)(2-py)]NO3 complexes (2-py is an anion of 2-pyridinecarboxylic acid) show I.D.50 values smaller than cisplatin whereas [M(bpy)(2-pyz)]NO3 complexes (2-pyz is an anion of 2-pyrazinecarboxylic acid) show I.D.50 values larger than cisplatin against P388 cancer cells.

A new complex [Pt(NH3)2(ddtc)]NO3.2H2O as a 1:1 electrolyte has been prepared. This was characterized by spectroscopic methods. The electronic absorption spectrum of this complex in water suggests that it has a square planar geometry. The infrared, 1H NMR and x-ray photoelectron spectroscopic studies suggest the bonding of ammonia molecules and diethyldithiocarbamate as bidentate ligand to platinum(II) in this complex. The 50% inhibition value of this complex against P388 lymphocytic leukemia cells is comparable with cisplatin. This complex interacts with calf thymus DNA by coordinate covalent bond.

Four new binuclear complexes of formula [M2(bipy)2(BAA)]Cl2 (where M is Pt(II) or Pd(II), bipy is 2,2'-bipyridine, and BAA is a dianion of meso-alpha-alpha'-diaminoadipic acid (DAA) or meso-alpha,alpha'-diaminosuberic acid (DSA) have been synthesized. These complexes have been characterized by chemical analysis and ultraviolet-visible, infrared, and 1H NMR spectroscopy. The mode of binding of ligands in these complexes has been ascertained by infrared and detailed 1H NMR spectroscopy. These complexes are 1:2 electrolyte in conductivity water. They have also been tested against P388 lymphocytic leukemia cells and their target is DNA molecules. [Pt2(bipy)2(DSA)]Cl2, [Pd2(bipy)2(DSA)Cl2, and [Pd2(bipy)2(DAA)]Cl2 show I.D.50 values comparable or lower than cis-diamminedichloroplatinum(II) and [Pt(bipy)(Ala)]Cl. In addition, binding studies of [Pt2(bipy)2(DSA)]Cl2 and [Pd2(bipy)2(DAA)]Cl2 to calf thymus DNA have been carried out and the mode of binding seems to be hydrogen bonding, as suggested earlier for analogous mononuclear amino acid-DNA complexes.

The effect of vitamin K3 (2-methyl-1,4-naphthoquinone) on Adriamycin (ADR) induced growth inhibition of drug sensitive and multidrug resistant P388 leukemia cells was evaluated. Exposure to ADR concentrations of 100-5000 ng simultaneously with 1 microM vitamin K3 elicited an enhanced inhibition of tumor cell survival. The effect of treatment with ADR alone, or in combination with vitamin K3 on DNA and RNA biosynthesis in the sensitive and resistant tumor cells, was also assessed. DNA and RNA biosynthesis inhibition was increased in P388/S (the parental cell line) and P388/ADR cells (the ADR resistant cell line which exhibits the multidrug resistant (MDR) phenotype) exposed to ADR after pretreatment for 3 h with vitamin K3. Concurrent administration in vivo of vitamin K3 and ADR illustrated a therapeutically significant increase (P less than 0.05) in the life span of sensitive and resistant tumor cell bearing animals. Vitamin K3 caused a depletion of the intracellular glutathione (GSH) levels in P388/S and P388/ADR leukemia cells but at concentrations greater than those that enhanced ADR cytotoxicity. Pretreatment of the tumor cells with 1 microM vitamin K3 induced a 35-50% (P less than 0.001) elevation in the intracellular ADR accumulation in MDR P388 leukemia cells, while such an effect was absent in P388/S tumor cells. DNA binding studies performed utilizing calf thymus DNA, indicated that vitamin K3 enhanced the intercalation potential of ADR and also altered the equilibrium between the free and bound form of ADR in a cell free system. These factors and their possible effects on the potentiation of ADR cytotoxicity and the therapeutic significance of utilizing vitamin K3 as an adjuvant in the chemotherapy of MDR tumors is discussed.

Several new L-amino acid derivatives of 2,2'-bipyridine and 1,10-phenanthroline complexes of platinum (Pt) and palladium (Pd) and a few binuclear 2,2'-bipyridine complexes of these metals were tested for their potential to inhibit rat hepatic nuclear transcription in vitro. Pd complexes were generally more effective inhibitors of transcription than the corresponding Pt complexes. Among Pd-diimine chlorides, the 2,2'-bipyridine complex was nearly 10 times more active than the corresponding 1,10-phenanthroline complex. Both Pt-diimine chlorides, however, showed same level of inhibitory activity. Amino acid derivatives were less inhibitory with respect to the parent metal diimine chlorides except for 1,10-phenanthroline complexes of Pd. For binuclear 2,2'-bipyridine complexes of Pt, the increase in length of linking hydrocarcon chain increased the inhibitory potential of the complex. The mechanism of inhibition of transcription by these metal complexes was sought to be understood by use of actinomycin-D and poly[d(I-C)] to differentiate effect on the two major components of transcription machinery viz. the template and the enzyme. These studies along with studies on reconstituted system of transcription using either pretreated template or enzyme indicate that these metal complexes displayed dual effect on transcription by inhibiting both the template and the enzymes.

Eleven new complexes of formula [M(NN)(XO3)] (where M is Pd(II) or Pt(II); NN is 2,2'-bipyridine, 1,10-phenanthroline, 2,2'-dipyridylamine, ethylenediamine or (+-)trans-1,2-diaminocyclohexane, and XO3(2-) is SeO3(2-) or TeO3(2-)) have been synthesized. These water soluble complexes have been characterized by chemical analysis and conductivity measurements as well as ultraviolet-visible and infrared spectroscopy. In these complexes the selenite or tellurite ligand coordinates to platinum(II) or palladium(II) as bidentate with two oxygen atoms. These complexes inhibit the growth of P 388 lymphocytic leukemia cells, their targets are DNA. The selenite complexes invariably show I.D.50 values less than cisplatin. However, the I.D.50 values of the tellurite complexes are usually higher than cisplatin, except that of [Pd(dach)(TeO3)] which has comparable I.D.50 values, as compared to cisplatin. [Pt(bipy)(SeO3)] and [Pd(bipy)(SeO3)] have been interacted with calf thymus DNA and bind to DNA through a coordinate covalent bond.

The syntheses of nine palladium(II) complexes of type [Pd(phen)(AA)]+ (where AA is an anion of glycine, L-alanine, L-leucine, L-phenylalanine, L-tyrosine, L-tryptophan, L-valine, L-proline, or L-serine) have been achieved. These palladium(II) complexes have been characterized by ultraviolet-visible, infrared, and 1H NMR spectroscopy. The binding studies of several complexes [M(NN)(AA)]+ (where M is Pd(II) as Pt(II), NN is 2,2'-bipyridine or 1,10-phenanthrodine, and AA is an anion of amino acid) with calf thymus DNA have been carried out using UV difference absorption and fluorescence spectroscopy. The mode of binding of the above complexes to DNA suggests the involvement of the hydrogen bonding between them. Several complexes [M(phen)(AA)]+ (where M is Pd(II) or Pt(II) and AA is an anion of amino acid) have also been screened for cytotoxicity in P388 lymphocytic cells. Of them, only two complexes, [Pd(Phen)(Gly)]+ and [Pd(phen)(Val)]+, show comparable cytotoxicity, as cisplatin does.

A series of complexes of the type [Pt(phen)(AA)]+ (where AA is the anion of glycine, L-alanine, L-leucine, L-phenylalanine, L-tyrosine, or L-tryptophan) has been synthesized. These complexes have been characterized by electronic absorption, infrared, and 1H NMR spectroscopy. The interaction of these complexes with calf thymus DNA has been studied using fluorescence spectroscopy. They inhibit the intercalation of ethidium bromide in DNA by intercalative binding at low concentrations and show nonintercalative binding at higher concentrations.

Eight new palladium complexes of the formula [Pd(bipy)(AA)]Cl 1 or 2 H2O (where bipy is 2,2'-bipyridine and AA is an anion of glycine, L-alanine, L-leucine, L-proline, L-serine, L-lysine, L-asparagine, or L-glutamine) have been synthesized by reaction of [Pd(bipy)Cl2] with an appropriate mono sodium salt of amino acid in water. These complexes have been characterized by chemical analysis and by visible, infrared, and 1H NMR spectroscopy. The detailed 1H NMR and infrared spectral studies of these complexes ascertain the mode of binding of amino acids to palladium through nitrogen of terminal -NH2 group and oxygen of terminal -COO- group. The molar conductance values of these complexes in water suggest them to be 1:1 electrolytes. These complexes have also shown growth inhibition against L1210 lymphoid leukemic, P388 lymphocytic leukemic, Sarcoma 180, and Ehrlich ascitic tumor cells. Some of these complexes show better 50% inhibitory dose values than cis-diamminedichloroplatinum(II).

Four new water soluble complexes of the formula [Pt(DA)(DDTC)]NO3 (where DA is 2,2'-bipyridine, 1,10-phenanthroline, 1,2-diaminopropane, or 1,2-diaminocyclohexane, and DDTC is diethyldithiocarbamate anion) have been synthesized by reaction of platinum-diamine/diimine diaqua complex with sodium diethyldithiocarbamate in molar ratio of 1:1. These complexes have been characterized by the chemical analysis, and ultraviolet-visible, infra-red and 1H NMR spectroscopy. The infrared and 1H NMR spectral studies of these complexes have ascertained the modes of binding of diamine/diimine and diethyldithiocarbamate to platinum. The molar conductance values of these platinum complexes in conductivity water suggest them to be 1:1 electrolytes. These four complexes and two other complexes containing ethylenediamine and 1,3-diaminopropane ligands have been tested against P-388 lymphocytic leukemic cells. Out of them only 2,2'-bipyridine and 1,10-phenanthroline complexes show 1.D.50 values less than cisplatin.

Resonance Raman spectra of mu-oxo-bis[iron(III) tetraphenylporphyrin] have been observed and compared to Raman spectra of the monomers iron(III) tetraphenylporphyrin chloride and bis(piperidine)iron(II) tetraphenylporphyrin. Selection rules for the Raman effect under conditions of resonance of the incident photons with electronic states are presented for the exact symmetry group (C2) and the pseudo-symmetry group (D4d): emphasis is placed on the physical processes involved in scattering via vibronic states of the dimer. These two models are experimentally distinguishable in the behavior of the depolarization ratios of the dimer vibrational doublets. Experimental data favor the assignment of D4d as the symmetry group relevant to a description of the various states of the dimer. The usefulness of resonance Raman spectra of the mu-oco-dimer of Fe(III) tetraphenylporphyrin has been discussed with respect to several biochemical systems where there is strong evidence of two or more closely interacting chromophores.

Seven platinum(II) complexes of the type [Pt(bipy)(AA)]n+ (where n = 1 or 0 and AA is anion of L-valine, L-isoleucine, L-aspartic acid (dianion), L-glutamic acid (dianion), L-glutamine, L-proline, or S-methyl-L-cysteine) have been prepared and characterized. The modes of binding of amino acids in these complexes have been ascertained particularly by infrared and 1H NMR spectral studies. The L-glutamine complex shows a ID50 value (50% inhibitory dose) in the range of greater than 20 micrograms/ml to 100 micrograms/ml of the complex. However, the complexes of L-valine, L-isoleucine, L-aspartic acid, L-glutamic acid, L-proline, and S-methyl-L-cysteine show ID50 values greater than 100 micrograms/ml of the complex. The above complexes also show inferior growth inhibition of P-388 cells than platinum(II) complexes of 2,2'-bipyridine with L-alanine, L-leucine, L-methionine, and L-aspargine as reported earlier. The platinum(II) complexes of 2,2'-bipyridine with glycine (Gly), L-alanine (Ala), L-leucine (leu), L-valine (Val), L-methionine (Met), L-phenylalanine (Phe), L-serine (Ser), L-tyrosine (Tyr) and L-tryptophan (Trp) have been tested for mutagenesis using TA 100 and TA 98 strains. They show nonmutagenicity. This is in contrast to the cis-[Pt(NH3)2Cl2] showing a base pair substitution mutagenesis.

Six new platinum complexes of the formula [Pt(2,2'-bipyridine)(amino acid)]n+, where n = 1 to 2 and amino acid is an anion of L-histidine, L-lysine, L-asparagine, L-phenylalanine, L-tryptophan, or L-tyrosine, have been prepared by interaction of [Pt(2,2'-bipyridine)Cl2] and an appropriate amino acid (sodium salt) in water or water-methanol mixture. They have been characterized by chemical analyses and spectral methods such as ultraviolet-visible, infrared, and 1H NMR spectroscopy. The 1H NMR studies of these complexes ascertain the modes of binding of amino acids to platinum. The histidine binds to platinum through the nitrogen of a -NH2 group and another nitrogen of heterocyclic ring. All other amino acids bind to platinum through nitrogen of a -NH2 group and oxygen of a -COO- group. The mode of binding of some amino acids to platinum in these complexes has been further confirmed by infrared spectroscopy, and the formulations of these complexes have been supported by conductivity measurements. These six amino acid complexes and also other complexes of glycine, alanine, leucine, serine, cysteine, methionine, and glutamine have shown growth inhibition against P-388 lymphocytic leukemic cells.

An important feature of Perutz's trigger mechanism for cooperativity in the reversible oxygenation of hemoglobin (Hb) is the tension along the histidine--metal linkage in deoxyHb and deoxycobaltohemoglobin (deoxy CoHb), supposedly due to the pull exerted by the globin on the metal atom. We have attempted to verify the existence of this pull by studying the emission Mössbauer spectra of deoxy 57CoHb and oxy 57 CoHb at different temperatures. The emission Mössbauer spectrum for none of the cobalt Hbs agrees with the absorption spectrum of the corresponding iron analog and, moreover, the spectrum of deoxy 57CoHb is characteristic of the intermediate-spin iron. These observations indicate that the daughter 57Fe atom is "frozen" almost in the same spatial situation as that of the parent 57Co. The protein is apparently holding the cobalt atom in position rather rigidly and, after the electron-capture decay of the 57Co atom, the protein does not permit the daughter 57Fe to move to a position characteristic of the iron atom.

The interactions of glycine, alanine, valine, leucine, proline, methionine, and asparagine derivatives of 2,2'-bipyridineplatinum(II) with calf thymus DNA, polyadenylic acid, and polyguanylic acid have been studied by difference absorption spectral technique. The association constants (Kapp) and number of binding sites per 100 nucleotides (n) have been obtained from binding isotherms which were constructed by treatment of data according to Scatchard equation. The Kapp values (2.2 X 10(4) to 4.6 X 10(5) M-1) of binding of platinum complexes with DNA are comparable to Kapp values (5.6 X 10(4) to 2.9 X 10(5) M-1) of binding of platinum complexes with polyguanylic acid. However, the polyadenylic acid does not exhibit any binding to platinum complexes. The hydrogen bonding is, however, the most probable mode of bonding involved in stabilizing the DNA-amino acid complexes.

The synthesis of virtually all the lanthanide octaethylporphyrin complexes have been achieved by heating appropriate anhydrous lanthanide halide and octaethylporphyrin in imidazole melt at 210 degrees C for two hours. The lighter lanthanide porphyrin complexes are very susceptible to hydrolysis, the middle lanthanide porphyrin complexes are moderately stable, and the heavier lanthanide porphyrin complexes are relatively more stable to hydrolysis. Two out of four lanthanide porphyrin complexes studied in detail, namely ytterbium and lutetium octaethylporphyrins, aggregate in benzene and the Soret bands in their absorption spectra are about 6 nm shifted to higher energies upon a hundred-fold increase in their concentrations. The aggregations of these lanthanide porphyrin complexes in non-coordinating solvents have been further verified by 1H NMR spectral studies. This spectral behavior can be interpreted qualitatively in terms of the model of the molecular exciton interactions with stacking of at least two prophyrins. A dimeric structure of these lanthanide porphyrin complexes has been proposed on the basis of geometrical considerations. On the contrary, the europium and gadolinium octaethylporphyrins associate very weakly in benzene in the concentration range studied. All four lanthanide porphyrin complexes interact with pyridine and piperidine, and the Soret bands in their absorption spectra are about 8 nm shifted to low energies as compared with their values in pure benzene.

Nine new palladium(II) complexes of the formula [Pd(bipy)(AA)]n+ (where bipy is 2,2'-bipyridine, AA is an anion of L-cysteine, L-aspartic acid, L-glutamic acid, L-methionine, L-histidine, L-arginine, L-phenylalanine, L-tyrosine, or L-tryptophan, and n = 0 or 1) have been synthesized by interaction of [Pd(bipy)Cl2] with an appropriate sodium salt of amino acid in water. These palladium(II) complexes have been characterized by chemical analysis and by visible, infrared, and 1H NMR spectroscopy. The modes of binding of amino acids in these palladium complexes have been ascertained by infrared and 1H NMR spectroscopy. The molar conductances of these complexes in water suggest that they are either nonelectrolytes or 1:1 electrolytes. These palladium complexes have shown growth inhibition against L1210 lymphoid leukemic, P388 lymphocytic leukemic, Sarcama 180, and Ehrlich ascites tumor cells. Some of these complexes show I.D.50 values comparable to or lower than cis-diamminedichloroplatinum(II).

A synthetic carbon monoxide complex of ruthenium (II) myoglobin has been reconstituted from apomyoglobin and the carbon monoxide of ruthenium (II) mesoporphyrin IX. This synthetic myoglobin complex shows an absorption spectrum with normal Soret and beta bands, and a split alpha band. The alpha-band splitting is not observed in the spectrum of the carbon monoxide derivative of ruthenium (II) mesoporphyrin IX in pyridine, even though the width of the alpha band in pyridine is narrower than in theprotein. The separation between two alpha bands in the spectrum of the protein is reduced from 7.5 to 6 nm in the presence of 2 M NaCl. This observation is interpreted in terms of perturbation of concentrated NaCl on the protein. The separation between the maxima of two alpha bands in the spectrum of the ruthenium (II) myoglobin also becomes smaller with decrease in pH from 7 to 4.5, and this process involves the distal histidine. The alpha band splitting in this protein complex is interpreted in terms of rhombic distortion of the square planar symmetry of the metal porphyrin in the protein.