Synthesis and Biological Properties of Enkephalin-like Peptides Containing Carboranylalanine in Place of Phenylalanine

The syntheses of [D -Ala², Car⁴, Leu⁵]-enkephalin, its amide, and of [D -Ala², Car⁴, Met⁵]-enkephalin amide are described in detail. The three compounds show pronounced morphine-like potencies in certain bio-assays, and were used to investigate the influence of electronic, steric and hydrophobic properties of the side-chains of amino acids no. 4 and 5 on opioid activity (see [13]).     

Synthesis of enantiopure, axially chiral, C-tetrasubstituted α-amino acids with binaphthyl-based crowned side chains and 3D-structural analysis of their peptides

The syntheses of the terminally protected, crowned, C^α-tetrasubstituted α-amino acids with only axial chirality, the two diastereomers Boc-(S)-Bip[(R)-Binol-22-C-6]-OMe and Boc-(R)-Bip[(R)-Binol-22-C-6]-OMe, and their respective enantiomers Boc-(R)-Bip[(S)-Binol-22-C-6]-OMe and Boc-(S)-Bip[(S)-Binol-22-C-6]-OMe, all derived from 2′,1′:1,2; 1″,2″:3,4-dibenzcyclohepta-1,3-diene-6-amino-6-carboxylic acid (Bip), were performed by bis-alkylation with cyclization of racemic (R+S)-Boc-[HO]₂-Bip-OMe, possessing two phenolic OH groups at the 6,6′-positions of the biphenyl frame of Bip, using (+)-(R)- and (−)-(S)-Binol[(OCH₂CH₂)₂OTs]₂ (2,2′-bis[5-tosyloxy-3-oxa-1-pentyloxy]-1,1′-binaphthyl), respectively, as the alkylating agent followed by chromatographic separation. Two series of terminally protected model peptides to the hexamer level, containing the (R)-Bip[(S)-Binol-22-C-6] residue at i and i+3 positions of the sequence, combined with either l-Ala or l-Ala/Aib, were synthesized by solution methods. Their 3D-structural analyses by FTIR absorption and NMR suggest that these peptides preferentially adopt folded secondary structures.     

68Ga-Labeled [Leu13ψThz14]Bombesin(7–14) Derivatives: Promising GRPR-Targeting PET Tracers with Low Pancreas Uptake

The gastrin-releasing peptide receptor (GRPR) is a G-protein-coupled receptor that is overexpressed in many solid cancers and is a promising target for cancer imaging and therapy. However, high pancreas uptake is a major concern in the application of reported GRPR-targeting radiopharmaceuticals, particularly for targeted radioligand therapy. To lower pancreas uptake, we explored Ga-complexed TacsBOMB2, TacsBOMB3, TacsBOMB4, TacsBOMB5, and TacsBOMB6 derived from a potent GRPR antagonist sequence, [Leu¹³ψThz¹⁴]Bombesin(7–14), and compared their potential for cancer imaging with [⁶⁸Ga]Ga-RM2. The K_i(GRPR) values of Ga-TacsBOMB2, Ga-TacsBOMB3, Ga-TacsBOMB4, Ga-TacsBOMB5, Ga-TacsBOMB6, and Ga-RM2 were 7.08 ± 0.65, 4.29 ± 0.46, 458 ± 38.6, 6.09 ± 0.95, 5.12 ± 0.57, and 1.51 ± 0.24 nM, respectively. [⁶⁸Ga]Ga-TacsBOMB2, [⁶⁸Ga]Ga-TacsBOMB3, [⁶⁸Ga]Ga-TacsBOMB5, [⁶⁸Ga]Ga-TacsBOMB6, and [⁶⁸Ga]Ga-RM2 clearly show PC-3 tumor xenografts in positron emission tomography (PET) images, while [⁶⁸Ga]Ga-TacsBOMB5 shows the highest tumor uptake (15.7 ± 2.17 %ID/g) among them. Most importantly, the pancreas uptake values of [⁶⁸Ga]Ga-TacsBOMB2 (2.81 ± 0.78 %ID/g), [⁶⁸Ga]Ga-TacsBOMB3 (7.26 ± 1.00 %ID/g), [⁶⁸Ga]Ga-TacsBOMB5 (1.98 ± 0.10 %ID/g), and [⁶⁸Ga]Ga-TacsBOMB6 (6.50 ± 0.36 %ID/g) were much lower than the value of [⁶⁸Ga]Ga-RM2 (41.9 ± 10.1 %ID/g). Among the tested [Leu¹³ψThz¹⁴]Bombesin(7–14) derivatives, [⁶⁸Ga]Ga-TacsBOMB5 has the highest tumor uptake and tumor-to-background contrast ratios, which is promising for clinical translation to detect GRPR-expressing tumors. Due to the low pancreas uptake of its derivatives, [Leu¹³ψThz¹⁴]Bombesin(7–14) represents a promising pharmacophore for the design of GRPR-targeting radiopharmaceuticals, especially for targeted radioligand therapy application.     

Sequencing of cyclosporins by fast atom bombardment and linked-scan mass spectrometry

The mass spectrometric sequence determination of amino acid residues in cyclosporins using fast atom bombardment, collisionally activated dissociations in the first field-free region and linked B/E scan is described. The general fragmentation scheme was derived from the spectra of cyclosporins A, B, C, D, F, G, L and [DH-MeBmt¹]CS. The main fragmentation pathways start by primary splitting between amino acids 2–3, 1–11 and 5–6. The corresponding N-terminal b-type ions are common fragment types in the mass spectra. The 1–11 splitting can be enhanced by the introduction of a lactone group into the peptide ring by conversion of cyclosporins into isocyclosporins. The fragmentation scheme was used for amino acid sequence determination in four new natural cyclosporins, [MeLeu¹]CS, [Leu⁴]CS, [Ile⁴]CS and [Leu⁵]CS.     

Opioid Peptides Determination by Tyrosinase-Modified Oxygen Electrode

Abstract

Opioid peptides morphiceptin (Tyr-Pro-Phe-Pro-NH₂), [D-Ala², D-Leu⁵]-enkephalin (Tyr-D-Ala-Gly-Phe-D-Leu, DADLE) and [D-Thr²]-Leu-enkephalin-Thr (Tyr-D-Thr-Gly-Phe-Leu-Thr, DTLET) containing tyrosine has been studied in the reaction with mushroom tyrosinase immobilized on Clark-type oxygen electrode. A 4–6-times higher response is observed for tyrosine compared to peptides. The detection limit of the tyrosinase-modified electrode for DADLE, morphiceptin and DTLET was 6 μM, 9 μM and 16 μM, respectively.     

The Synthesis of [4-Carboranylalanine, 5-Leucine]-Enkephalin (Including an Improved Preparation of t-Butoxycarbonyl-L-o-carboranylalnine,New Derivatives of L-Propargylglycine,and a Note on Melanotropic and Opiate Receptor Binding Characteristics)

The title compound, an analogue of [Leu⁵]-enkephalin with L -o-carboranylalanine replacing L -phenylalanine in position 4, was prepared by fragment condensation. The analogue has a 3-fold higher affinity for rat brain opiate receptors in the [³H]naloxone competition assay than natural [Leu⁵]-enkephalin. Like [Leu⁵]-enkephalin and N^a-acetyl-[Leu⁵]-enkephalin, the N-terminal tripeptide fragment, H · Tyr-Gly-Gly · OH, had no melanotropic activity in the Rana pipiens frog skin assay. A convenient, direct synthesis of methyl t-butoxycarbonyl-L -propargylglycinate is described, and the ¹³C-NMR. spectra of L -o-carboranylalanine recorded. The procedure was extended to the preparation of BOC · Car-Leu · OMe from BOC · Pra-Leu · OMe. A number of new propargylglycine derivatives are reported.     

Comparison of Crystal Structures of New Racemic Chiral Compounds Showing and Not Showing the Phenomenon of Preferential Enrichment

Abstract

The crystal structures of (±)-[2-[4-(3-ethoxy-2-hydroxypropoxy)phenylcarbamoyl]ethyl] trimethylammonium p-chlorobenzenesulfonate [(±)-NCMe₃] and its terminal methoxy derivative, (±)-NCMe₃-OMe, are compared. The former racemate exhibited the phenomenon of Preferential Enrichment, whereas the latter failed to do so. Crystal data, (±)-NCMe₃: CuKα radiation, space group P 1, Z = 2, a = 9.848(5), b = 14.823(3), c = 9.147(1) Å, α = 97.81(1), β = 92.68(3), γ = 105.92(2)°, D _calc = 1.355 g/cm³, R = 0.056 for 3213 observed reflections; (±)-NCMe₃-OMe: CuKα radiation, space group P 1, Z = 2, a = 11.350(1), b = 14.568(2), c = 8.2981(4) Å, α = 94.346(7), β = 112.376(5), γ = 78.622(9)°, D _calc = 1.343 g/cm³, R = 0.069 for 1519 observed reflections.     

Membrane Structure of Substance P. II. Secondary Structure of Substance P, [9-Leucine]substance P,and Shorter Segments in 2,2,2-Trifluoroethanol,Methanol, and on Liposomes Studied by Circular Dichroism

Comparative CD studies with substance P ( 1 ), [Leu⁹]substance P ([Leu⁹]- 1 ), and their shorter peptide segments supported the membrane structures predicted for substance P and [Leu⁹]substance P. They indicated that the C-terminal segments (from residue 3 or 4 onward) can adopt α-helical conformations in hydrophobic environments and on lipid membranes. The N-terminal segment, (residues 1–4) had a poly(proline)-like conformation in aqueous and hydrophobic surroundings. Residues 3 and 4 (Lys-Pro) appeared to belong to both domains and bring about the transition between the two. The estimated free energies of transfer for 1 and [Leu⁹]- 1 from their random conformations in H₂O to their partially helical conformations on an aqueous-hydrophobic interface are too small to allow detectable interaction with neutral lipid membranes at low concentrations. The two peptides should, however, interact detectably with anionic membranes because of favourable Boltzmann distribution factors. This prediction was shown to be correct for liposomes prepared from 1,2-dioleoyl-sn-glycero-3-phosphocholine (neutral) and phosphatidylserine (anionic).     

Addition of iodine-based electrophilic reagents to some vinylsilanes

The addition of some Iodine-based electrophillic reagents to the cyclic vinylsilanes (1) and (4,R=Bu^t,R¹=H) and (4,R=H,R¹-OMe) occurs with high regio-and stereo-specificity to give (2,X=OMe,N₃, and NCS) and (5,R=Bu^t,R¹=H,X=OMe and N₃) and (5, R=H,R¹-OMe,X-OMe). Similar additions to 2-trimethylsilylhept-1-ene (6) are less efficient and occur with lower regioselectivity. Some of the adducts (2) and (5) have been transformed into functionalised organosilanes. Compounds (10,X=OMe,N₃, and NCS) and (12)–(15) are thereby obtained.     

Synthesis,structure, EPR,and DFT calculation on dinuclear paddle wheel Cu(II) complexes with bis-chelate rings

A series of diester-dicarboxylic acids, L¹H₂, L²H₂, L³H₂, L⁴H₂, and L⁵H₂ and their dinuclear Cu₂ complexes [Cu(L¹)CH₃CN]₂ (1), [Cu(L²)H₂O]₂ (2), [Cu(L³)CH₃CN]₂ (3), [Cu(L⁴)EtOH]₂ (4), and [Cu(L⁵)CH₃CN]₂ (5), were synthesized. The crystal structures obtained for 1, 2, and 4 and the density functional theory optimized structures for 2, 3, and 5 illustrated the formation of tetracarboxylate “paddle wheel” complexes. The phthalyl and diphenyl head groups and the spacer moieties were appropriately altered and the size of the chelate ring expanded from 15-membered in 1 to 21-membered in 5. The dinuclear units have strong Cu–Cu interaction with EPR spectra exploring spin coupled features.     

Central-to-axial chirality transfer and induced circular dichroism in 6,7-dihydro-5H-dibenz[c,e]azepine derivatives of α- and β-amino esters

DAZ-Xaa^∗-OMe amino ester derivatives with Xaa^∗ = d/l-Ala, d/l-Val, l-Leu, l-Ile, l-Ser, l-β³-HAla, l-β³-HVal, l-β³-HLeu, (1S,2S)/(1R,2R)-ACHC (2-aminocyclohexanecarboxylic acid) and (1S,2S)/(1R,2R)-ACPC (2-aminocyclopentanecarboxylic acid), N-blocked as 6,7-dihydro-5H-dibenz[c,e]azepines (DAZ), have been synthesized and evaluated for the determination of the absolute configuration of α- and β-amino esters through the induced circular dichroism of the biphenyl chromophore.     

Crystal structures,electrochemical properties,antioxidant screening and in vitro cytotoxic studies on four novel Cu(II) complexes of bidentate Schiff base ligands derived from 2-methoxyethylamine

Four novel Schiff base ligands and their copper(II) complexes, [Cu(L¹)₂] (1), [Cu(L²)₂] (2), [Cu(L³)₂] (3), and [Cu(L⁴)₂] (4), were synthesized and characterized by elemental analyses, FT-IR, and UV–Vis spectroscopy. The ligands were synthesized from the condensation of 2-methoxyethylamine with various salicylaldehyde derivatives (x-salicylaldehyde for HLⁿ, x = H (n = 1), 5-Br (n = 2), 3-OMe (n = 3), and 4-OMe (n = 4)). The molecular structures of 1, 2, and 3 were determined by the single crystal X-ray diffraction technique. The redox behavior studies of the complexes in acetonitrile display the electronic effects of the groups on the redox potential. The antioxidant activity of the Schiff base ligands and their Cu(II) complexes was evaluated using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging method and FRAP assay. Furthermore, the in vitro anticancer activity of compounds was screened, including MTT and migration assays against gastric cancer cell line (MKN-45). The results show that all ligands and complexes have antioxidant and anticancer activity in a concentration-dependent way.     

Polymeric organosilicon systems 14. Synthesis and some properties of alternating polymers composed of a dithienylene group and a mono-, di- or tri-silanylene unit

Poly[5, 5′ - (dimethylsilylene) - 2, 2′ - dithienylene] (4a), poly[5, 5′ -(methylphenylsilylene)-2, 2′ -dithienylene] (4b), poly[5, 5′ -(1, 1, 2, 2-tetramethyldisilanylene)-2, 2′ -dithienylene] (4c), poly[5, 5′-(1, 2-dimethyl-1, 2-diphenyldisilanylene)-2, 2′ -dithienylene] (4d), poly[5, 5′-(1, 2, 2, 2-tetramethyldisilanylene)-2, 2′-dithienylene] (4e), and poly[5, 5′-(1, 1, 2, 2, 3, 3 - hexamethyltrisilanylene) - 2, 2′ -dithienylene] were synthesized by dehalogenative coupling of the respective bis(2-bromothienyl)- substituted mono, di- and tri-silanes with magnesium in the presence of a catalytic amount of a nickel(II) complex in 16–99% yields. The polymers thus obtained are light-yellow solids and soluble in common organic solvents. Molecular weights, M_w, of the polymers were measured and found to be 7800–35 000 by gel-permeation chromatography relative to polystyrene standards. The photochemical properties of the polymers (4a–4d) having silylene and disilanylene units were investigated. Only poly[5, 5′-(1, 2-dimethyl-1, 2-diphenyl-disilanylene)-2, 2′-dithienylene] (4d) was found to be photoactive, but the others were inactive. When the thin solid films prepared from the polymers 4a–4e by spin-coating were exposed to antimony(V) fluoride in vacuo, the films became conducting; their conductivities were determined to be 10^?2 – 10^?3 S cm^?1 by the four-probe method.     

Evaluation of the binding ability of tetraaza[2]arene[2]triazine receptors anchoring l-alanine units for aromatic carboxylate anions

The binding affinities of three new tetraaza[2]arene[2]triazine based macrocycles anchoring one (AC1A) and two (AC2A and Me₄AC2A) l-alanine amino acid units for five aromatic carboxylate anions (bz^?, ph^2?, iph^2?, tph^2? and btc^3?) were investigated in DMSO-d₆. ¹H NMR titrations revealed that the AC1A and AC2A receptors exhibit comparable anion affinities, suggesting that the two l-alanine binding units in AC2A are not simultaneously involved in the anion recognition as indicated by molecular dynamics simulations carried out for selected AC1A and AC2A associations using the AMBER force field (GAFF). New force field parameters were developed in order to mimic the structural singularity derived from the N–H macrocyclic bridges.     

Naphthalene and Anthracene Complexes Sandwiched by Two {(Cp*)FeI} Fragments: Strong Electronic Coupling between the FeI Centers

The reactions of the half‐sandwich iron(II) complex [FeCl(Cp*)(tmeda)] ( 1 ; Cp*=η⁵‐C₅Me₅, TMEDA=N,N,N′,N′‐tetramethylethylenediamine) with potassium naphthalenide or potassium anthracenide gave the diamagnetic complexes [(Cp*)Fe(μ‐polyarene)Fe(Cp*)] (polyarene=naphthalene ( 2 ), anthracene ( 3a )), which have two {(Cp*)Fe} units bound to opposite faces of the polyarene. One of two {(Cp*)Fe} units in 3a is located over the central ring of anthracene while the other is positioned over an outer ring. The {(Cp*)Fe} unit bound to the central ring of 3a migrates to the outer ring upon heating in the solid state to give the isomer 3b . The electrochemical potential separations between successive one‐electron redox events for complexes 2 and 3b are large. The mixed valence complexes [ [2]+ ]⁺ and [ [3b]+ ]⁺ were synthesized by chemical oxidation. The mixed‐valence complex [ [3b]+ ]⁺ is charge delocalized on the Mössbauer timescale at 78 K, and its absorption spectrum shows an intervalence charge‐transfer band. Complex [ [2]+ ]⁺ exhibits two absorption bands in the near‐IR region and a slightly broadened doublet in the Mössbauer spectrum. DFT calculations were carried out to examine the electronic structures of these dinuclear iron(I) complexes to elucidate the factors responsible for their diamagnetism and to determine the degree of charge delocalization in the mixed‐valence complexes.     

Biosynthetic pathway of 24-membered macrolactam glycoside incednine

Incednine was isolated from Streptomyces sp. ML694-90F3 as an inhibitor of anti-apoptotic function of Bcl-2/Bcl-xL oncoproteins. The structure of incednine is quite unique with a characteristic 24-membered macrocyclic lactam aglycone and two unusual aminosugars. To understand its biosynthetic pathway, the incorporation studies were carried out with [1-¹³C]acetate, [1,2-¹³C₂]acetate, [1-¹³C]propionate, l-[¹³C₅,¹⁵N]glutamate, [1,2,3-¹³C₃]glycerol, d-[6,6-²H₂]glucose, and l-[CH₃-¹³C]methionine. As a result, acetate, propionate, and glycerol were well incorporated into the elongation units of the macrolactam moiety, which indicates that its basic skeleton could be constructed by standard polyketide synthase, whereas all atoms of the starter unit were labeled by [¹³C₅,¹⁵N]glutamate suggesting that glutamate is somehow decarboxylated and rearranged into 3-aminobutyrate as the unique starter unit. The origins of the sugar moieties and methyl groups were also clarified. Based on the incorporation pattern, a plausible biosynthetic pathway for incednine is proposed.     

(−)-Ternatin, a highly N-methylated cyclic heptapeptide that inhibits fat accumulation: structure and synthesis

A highly N-methylated cyclic heptapeptide, (−)-ternatin, was isolated from the mushroom Coriolus versicolor, which significantly suppressed fat accumulation against 3T3-L1 murine adipocytes (EC₅₀ = 0.14 μg/mL). Although ternatin was previously reported to be antibacterial or antimicrobial compound, its inhibitory effect on fat accumulation has been first shown. The structure of (−)-ternatin was revised to be a cyclo [d-allo-Ile¹-l-(NMe)Ala²-l-(NMe)Leu³-l-Leu⁴-l-(NMe)Ala⁵-d-(NMe)Ala⁶-(2R,3R)-3-hydroxy-Leu⁷] by spectroscopic analysis and chemical synthesis.     

Thiocyclosporins: Preparation,Solution and Crystal Structure,and Immunosuppressive Activity

The reaction of cyclosporin A (CsA) with Lawesson's reagent under different conditions yields various thiocyclosporins, in which carbonyl O-atoms and/or the hydroxy O-atom of the MeBmt residue are replaced by an S-atom. The position of the S-atom is determined by NMR spectroscopy, and the conformations of the products are studied by NMR spectroscopy and X-ray crystallography. Some of the thiocyclosporins show interesting conformational properties. Whereas one conformation strongly dominates for CsA in CDCL₃, two conformers A and B, in a ratio 58:42 are found for [¹ψ², CS–NH]CsA. Extensive NMR studies including new 2D and 3D heteronuclear techniques and restrained MD calculations using ROE effects demonstrate that the major conformer A is identical to CsA, while the minor conformer B contains an additional cis peptide bond between the Sar³ and MeLeu⁴ residues. [⁴ψ⁵, CS? NH; ⁷ψ⁸, CS–NH]CsA exhibits a conformation very similar to crystalline CsA. However, the D-Ala⁸NH, MeLeu⁶Co γ-turn H-bond is not present in this dithio analogue. Also different is the MeBmt¹side-chain conformation, the dithio conformation showing a strong MeBmt¹OH, Sar³CO H-bond. Immunosuppressive activities of thiocyclosporins are measured in IL-2 and IL-8 reporter gene assays. Their activities are discussed in relation to their conformations.     

Structural preferences of two unnatural hybrid octapeptides with and without the crystal environment: a computational study

A computational study by combining molecular dynamics simulations with density functional theory calculations has been conducted on two unnatural octapeptides, Boc-[Aib-β³(R)Val]₄-OMe and Boc-[Aib-α(S)Val]₄-OMe, and the corresponding infinite one-dimensional chains to understand their inherent structural preferences and the effect of the crystal environment on their structures. For both systems, we have determined their gas-phase lowest energy conformers, which possess hybrid helical structures (not pure α-helix-like or 3₁₀-helix-like). For Boc-[Aib-β³(R)Val]₄-OMe, our calculations show that the corresponding infinite polypeptide has an α-helix-like structure as the most stable structure. This result is in accord with the experimental crystal structure. For Boc-[Aib-α(S)Val]₄-OMe, our calculations show that the infinite polypeptide will exist in 3₁₀-helix-like structure, which is also consistent with the observed crystal structure. The present study demonstrates that the peptide conformer in the crystal environment does not necessarily correspond to the most stable structure in vacuum.