New Ecotoxicologically and Biogenetically Relevant Terpenes of the tropical green seaweed Caulerpa taxifolia which is invading the Mediterranean

The tropical green seaweed Caulerpa taxifolia (VAHL ) C. AGARDH (Caulerpales) which is invading the Mediterranean is shown to contain trace amounts of two further novel terpenes, 7,7-C-didehydro-6-hydroxy-6,7-dihydrocaulerpenyne (= (4S, 6S,1E)-3-[(Z)-acetoxymethylidene]-6-hydroxy-11-methyl-7-methylidenedodeca-1,10-dien-8-yne-1,4-diyl diacetate; 3a ) and taxifolione (= 6-methylhept-5-en-3-yn-2-one; 4 ). The former is the most active of the toxins so far isolated from this seaweed, both as an in vitro inhibitor of the growth of marine bacteria and as a cytotoxic agent toward marine ciliate protists. This suggests a central ecotoxicological role for triacetate 3a as an adjuvant factor in the invasion of the Mediterranean by this seaweed. Moreover, the almost equally toxic 10,11 -epoxycaulerpenyne ( 2 ) which is scarcely available from Nature for bioassays can now be obtained by peroxy-acid epoxidation of caulerpenyne ( 1 ), along with the 6,7-epoxycaulerpenynes 6b and 6a . The latter are very labile, 6a giving triacetate 3a , suggesting epoxides to be late biogenetic intermediates in C. taxifolia.     

Raspailynes,Novel Long-Chain Acetylenic Enol Ethers of Glycerol from the Marine Sponges Raspailia pumila and Raspailia ramosa

The sponges Raspailia pumila and ramosa (Demospongiae, Tetractinomorpha, Axinellida) from the North-East Atlantic are shown to contain a series of novel long-chain enol ethers of glycerol where the enol ether C?C bond is conjugated, in sequence, to both an acetylenic and an olefinic bond. Polar extracts give raspailynes hydroxylated at their (1Z5Z)-1,5-alkadien-3-ynyl chain, like raspailyne Al ( = (+)-(S)-3-[((1Z,5Z)-16-hydroxy-hexadeca-1,5-dien-3-ynyl)oxy]-1,2-propanediol; (+ 2 ) and isoraspailyne A ( = (+)-3-[((1Z,5Z)-17-hydroxyocta-deca-1,5-dien-3-ynyl)oxy]-1,2-[propanediol; (+)- 3 ). Less polar extracts give 3 different types of raspailynes not hydroxylated at the chain. Raspailynes of the first type have either the (1Z,5Z)-configuration in a linear chain such as raspailyne B2 (( = (?)-(s)-3-[((1Z,5Z)-trideca-1,5-dien-3-ynyl)oxy]-1,2-propanediol; (?)-4), raspailyne Bl ( = (?)-3-[((1Z,5Z)-tetradeca-1,5-dien-3-ynyl)oxy]-1,2-propanediol;(?)- 5 ), and raspailyne B ( = 3-[((1Z,5Z)-pentadeca-1,5-dien-3-ynyl)oxy]-1,2-propanediol; 6 ) or the (1Z,5Z)-pentadeca-1,5-dien-3-ynyl)oxy]-1,2-propanediol; 6 )or the (1Z,5Z)-configuration in a chain ending with an isopropyl group, like isoraspailyne Bl ( = 3-[((1Z,5Z)-12-methyltrideca-1,5-dien-3-ynyl)oxy]-1,2-propanediol; 7 ) and isoraspailyne B ( = 3-[((1Z,5Z)-13-methyltetradeca-1,5-dien-3-ynyl)oxy]-1,2-propanediol; 8 ). Raspailynes of the second type have the (1Z,5E)-configuration, like isoraspailyne Bla ( =3-[((1Z,5E)-tetradeca-1,5-dien-3-ynyl)oxy]-1,2-propanediol; 9 ) and isoraspailyne Ba ( = 3-[((1Z,5E)-13-methyltetradeca-1,5-dien-3-ynyl)oxy]-1,2-propanediol; 10 ). Raspailynes of the third type have the (1E,5Z)-configuration, like isoraspailyne Blb ( = 3-[((1E,5Z)-tetradeca-1,5-dien-3-ynyl)oxy]-1,2,-propanediol; 11 ). The (S)-configuration for (+)- 1 ,((+)- 2 , and (?)- 4 is derived from chemical correlations.     

Synthesis of kairomones and their analogs from 2-acylcyclohexane-1,3-diones with an unsaturated acyl side chain

Natural 2-[(9Z)-1-oxooctadec-9-en-1-yl]-, 2-[(9Z,12Z)-1-oxooctadeca-9,12-dien-1-yl]-, and 2-[(9Z,12Z,15Z)-1-oxooctadeca-9,12,15-trien-1-yl]cyclohexane-1,3-diones (components of flour moth Ephestia kuehniella kairomones) and some their analogs were synthesized from cyclohexane-1,3-diones and long-chain unsaturated carboxylic acid chlorides.     

1-R-2-[(1E,3E)-4-Aminobuta-1,3-dien-1-yl]-1H-benzimidazoles. Synthesis and some transformations

7-Nitropyridobenzimidazolium salts are cleaved with secondary amines to form 2-[(E,E)-4-aminobuta-1,3-dienyl]-1H-benzimidazoles. The latter react with dimethyl acetylene-dicarboxylate to yield 4a-[(E,Z,E)-6-amino-4,5-dimethoxycarbonylhexa-1,3,5-trien-1-yl]-1,2,3,4-tetra(methoxycarbonyl)-4a,5-dihydropyrido[1,2-a]benzimidazoles.     

Pd(II)-mediated coupling of 1,1-dimethyl-2-propynyl acetate with itself and 1-decene

Heating 1,1-dimethyl-2-propynyl acetate (1) with PdCl₂(MeCN)₂ (0.05–0.1 mol equiv) and MeCN alone afforded (E)-2,7-dimethylocta-2,4,6-trien-3,6-diyl diacetate (2) (yield ca. 20%) and with 1-decene (7 mol equiv) present (E)-2-methyltetradeca-2,4-dien-3-yl acetate (7) (30%) together with 2 (12%).     

Formal synthesis of strobilurins A and X

Known synthetic precursors of strobilurins A and X, i.e., methyl (3Z,5E)-6-aryl-3-methylhexa-3,5-dienoates (aryl is phenyl, 4-methoxyphenyl), were synthesized by highly stereospecific reactions from 2-(2-tert-butyldimethylsilyloxyethyl)- and 2-[2-(4-methoxybenzyloxy)-ethyl]-5-arylpenta-2E,4E-dien-1-ols. These dienols were efficiently dehydroxylated to (1E,3Z)-4-methyl-6-(4-methoxybenzyloxy)hexa-1,3-dienylarenes with their subsequent demethoxyben-zylation to (3Z,5E)-6-aryl-3-methylhexa-3,5-dien-1-ols. The latter through the step of corresponding aryldienals and aryldienoic acids were transformed to the target methyl (3Z,5E)-6-aryl-3-methylhexa-3,5-dienoates, which completes a formal synthesis of strobilurins A and X. Configuration of the C=C bonds of the conjugated aryldiene system is preserved in the considered transformations by 95–97%.     

Stereocontrolled synthesis of (3Z,5E)-6-aryl-3-methylhexa-3,5-dien-1-ols,intermediates in the synthesis of strobilurin antibiotics

(2E,4E)-5-Aryl-2-(2-benzyloxyethyl)penta-2,4-dien-1-als (aryl is phenyl and 4-methox-yphenyl) were reduced with NaBH₄ quantitatively and stereospecifically to the corresponding penta-2(E),4(E)-dien-1-ols. The hydroxymethyl group in the latter was transformed into a methyl one with a stereoselectivity of 92–97%. Debenzylation of the resulting (1E,3Z)-1-aryl-6-benzyloxy-4-methylhexa-1,3-dienes with AlCl₃ in the presence of PhNMe₂ afforded the target (3Z,5E)-6-aryl-3-methylhexa-3,5-dien-1-ols; the configuration of the C=C bonds in the conjugated aryl diene systems was retained at 95%.     

Caulerpenyne-Amine Reacting System as a Model for in vivo Interactions of Ecotoxicologically Relevant Sesquiterpenoids of the mediterranean-adapted tropical green seaweed caulerpa taxifolia

Caulerpenyne ( 1 ), the most abundant of the ecotoxicologically relevant sesquiterpenoids of the Mediterranean-adapted tropical green seaweed Caulerpa taxifolia, was found to react with Et₃N or pyridine in MeOH by initial deprotection of C(1)HO to give oxytoxin 1 ( 2a ), previously isolated from the sacoglossan mollusc Oxynoe olivacea. With BuNH₂, without any precaution to exclude light, 1 gave the series of racemic 3 and 4 , and achiral (4E,6E)- 5 , (4E,6Z)- 5 , (4Z,6E)- 5 , and (4Z,6Z)- 5 pyrrole compounds, corresponding to formal C(4) substitution, 4,5-β-elimination, and (E/Z)-isomerization at the C(4)?C(5) and C(6)?C(7) bonds. Changing to CDCl₃ as solvent in the dark, 1 gave cleanly, via 2a as an intermediate, 3 and (4E,6E)- 5 . The latter proved to be prone to (E/Z)-photoisomerization. Under standard acetylation conditions, 3 gave (4E,6E)- 5 via acetamide 7 as an intermediate. Particular notice is warranted by selective deprotection of 1 at C(1), mimicking enzyme reactions, and unprecedented formation of pyrrole compounds from freely-rotating, protected 1,4-dialdehyde systems.     

The Photoisomerization Behavior of the (4-Hydroxycinnamoyl)-spermidines

The photoisomerization behavior of three mono[(E)-3-(4-hydroxyphenyl)prop-2-enoyl]spermidines, 1, 2 , and 3 , and three bis[(E)-3-(4-hydroxyphenyl)prop-2-enoyl]spermidines, 4, 5 , and 6 , are investigated. The synthetic product (E)- 1 could be almost quantitatively (> 96%) converted into its isomer (Z)- 1 under UV light irradiation. In the cases of (E)- 2 and (E)- 3 , a mixture of (E)/(Z) ca. 1:2 was obtained, when the same conditions were applied. The comparison of their UV spectra provides the possible explanation for these different behaviors. Furthermore, it was noticed that the (Z) → (E) isomerization of the C?C bond took place during the purification by reverse-phase high-performance liquid chromatography (RP-HPLC), and the (E)/(Z)-mixture is thus inseparable. The same feature could be observed during the isolation of the (Z,Z)-N,N′-bis[3-(4-hydroxyphenyl)prop-2-enoyl]-spermidines, (Z,Z)- 4 , (Z,Z)- 5 , and (Z,Z)- 6 . Nevertheless, the fractions of (Z,Z)- 5 and (Z,Z)- 6 were in almost pure state collected, and their ¹-NMR spectra are presented.     

Face Selectivity of the Diels-Alder Additions of Exocyclic Dienes Grafted onto 7-Oxabicyclo[2.2.1]heptanes

Stereoselective syntheses of 2exo, 3exo-bis (chloromethyl)-5-[(Z)-chloromethylidene]- ( 9 ), 2exo, 3exo-bis (chloromethyl)5-[(E)-chloromethylidene]- ( 10 ) and 2exo, 3exo-bis(chloromethyl)-5-[(E)-methoxymethylidene]-6-niethylidene-7-oxa-bicyclo[2.2.1]heptane ( 13 ) are presented. Double elimination of HCI from 9, 10 and 13 yielded 2-[(Z)-chloromethylidene]- ( 14 ), 2-[(E)chloromethylidene]- ( 15 ) and 2-[(E)-methoxymethylidene]-3,5,6-mmethylidene-7-oxabicycio[2.2.1]heptane ( 18 ), respectively, without loss of the olefin configuration. Ethylene tetracarbonitrile (TCE) and N-phenyltriazolinedione (NPTAD) added to these new exocyclic dienes and tetraenes preferentially onto their exo-face. The same face selectivity was observed for the cycloadditions of TCE to the (Z)- and (E)-chlorodienes 9 and 10 , thus realizing a case where the kinetic stereoselectivity of the additions is proven not to be governed by the stability of the adducts. The exo-face selectivity of the Diels-Alder additions of dienes grafted onto 7-oxabicyclo [2,2.1]heptanes contrasts with the endo-face selectivity reported for a large number of cycloadditions of dienes grafted onto bicyclo[2.2.1]heptane skeletons.     

Experimente zum kompetitiven Einbau der Stereoisomeren des Farnesols in Cantharidin. 6. Mitteilung zur Biosynthese des Cantharidins

Experiments on the competitive incorporation of farnesol-stereoisomers into cantharidin Farnesol ( 2 ) has been demonstrated to be an efficient precursor for cantharidin ( 1 ), into which it is transformed by elimination of C(1), C(5), C(6), C(7) and C(7′) [1]. The following incorporation experiments with doubly labelled (³H and ¹⁴C) stereoisomers of farnesol present strong evidence that (E,E)- farnesol ((E,E)- 2 ) in fact is the precursor for cantharidin, whereas (2E, 6Z)- 2 and (Z,Z)- 2 are not utilized for the biosynthesis of cantharidin. A possible mechanism for the incorporation of (2Z,6E)-farnesol ((2Z,6E)- 2 ) to an extent of 56,8% relative to (E,E)- 2 is discussed.     

A bioactive cycloartane triterpene from Garcinia hombroniana

The dichloromethane bark extract of Garcinia hombroniana yielded one new cycloartane triterpene; (22Z,24E)-3β-hydroxycycloart-14,22,24-trien-26-oic acid (1) together with five known compounds: garcihombronane G (2), garcihombronane J (3), 3β acetoxy-9α-hydroxy-17,14-friedolanostan-14,24-dien-26-oic acid (4), (22Z, 24E)-3β, 9α-dihydroxy-17,14-friedolanostan-14,22,24-trien-26-oic acid (5) and 3β, 23α-dihydroxy-17,14-friedolanostan-8,14,24-trien-26-oic acid (6). Their structures were established by the spectral techniques of NMR and ESI-MS. These compounds together with some previously isolated compounds; garcihombronane B (7), garcihombronane D (8) 2,3’,4,5’-tetrahydroxy-6-methoxybenzophenone (9), volkensiflavone (10), 4’’-O-methyll-volkensiflavone (11), volkensiflavone-7-O-glucopyranoside (12), volkensiflavone-7-O-rhamnopyranoside (13), Morelloflavone (14), 3’’-O-methyl-morelloflavone (15) and morelloflavone-7-O-glucopyranoside (16) were evaluated for cholinesterase enzymes inhibitory activities using acetylcholinesterase and butyrylcholinesterase. In these activities, compounds 1–9 showed good dual inhibition on both the enzymes while compounds 10–16 did not reasonably contribute to both the cholinesterases inhibitory effects.     

Synthesis of the Bretonins,Polyolefinic Esterified Glyceryl Ethers of an Unidentified Sponge from the North-Brittany Sea: Absolute Configuration and Novel Structure Assignment

The absolute configurations of acetylated bretonin A (= (+}-( R )-1-[(acetoxy)methyl]-2-{[(4E,6E,8E)-dodeca-4,6,8-trienyl]oxy}ethyl 4-acetoxybenzoate; (?)- 1b ) and isobretonin A (= (+)-(S)-3-{[(4E,6E,8E)-do-deca-4,6,8-trienyl]oxy}-2-hydroxypropyl 4-hydroxybenzoate; (+)-2), previously isolated from an undetermined sponge of the North Brittany sea, were established by comparison with synthetic (+)- lb and (+)- 2 , obtained from the condensation of commerical (?)-(R)-2,2-dimethyl-1,3-dioxolan-4-yl p-toluenesuifonate ((?)-(R)- 15 ) with a mixture of (4E,6E,8E)- ( 14e ) and (4E,6Z,8E)-dodeca-4,6,8-trien-1-ol ( 14z ). This also allowed confirming the structure and configuration of bretonin B (= (S)-2-{[(4E,6Z,8E)-dodeca-4,6,8-trienyl]oxy}-1-(hydroxy-methyl)ethyl 4-hydroxybenzoate; 3 ) which was also isolated from the same sponge, albeit in a too small amount for a complete study. As concerns the glyceryl ethers precursors of the bretonins, co-occurrence of the usual (S)-con-figuration (from 1a ) with the unusual (R)-configuration (from (+)- 2 )) poses intriguing biogenetic problems.     

Allgemeiner Zugang zu Polyaminen mit Ethan-1,2-diamin-Einheiten: Synthese von nicht natürlich vorkommenden homologen und isomeren N1,4-Di(4-cumaroyl)sperminen

█tl="American"█The synthesis of the three N,N′-di(4-coumaroyl)tetramines, i.e., of (E,E)-N-{3-[(2-aminoethyl)amino]propyl}-3,3′-bis(4-hydroxyphenyl)-N,N′-(ethane-1,2-diyl)bis[prop-2-enamide] ( 1a ), (E,E)-N-{4-[(2-aminoethyl)amino]butyl}-3,3′-bis(4-hydroxyphenyl)-N,N′-(ethane-1,2-diyl)bis[prop-2-enamide] ( 1b ), and (E,E)-N-{6-[(2-aminoethyl)amino]hexyl}-3,3′-bis(4-hydroxyphenyl)-N,N′-(ethane-1,2-diyl)bis[prop-2-enamide] ( 1c ), is described. It proceeds through stepwise construction of the symmetric polyamine backbone including protection and deprotection steps of the amino functions. Their behavior on TLC in comparison with that of 1,4-di(4-coumaroyl)spermine (=(E,E)-N-{4-[(3-aminopropyl)amino]butyl}-3,3′-bis(4-hydroxyphenyl)-N,N′-(propane-1,3-diyl)bis[prop-2-enamide]; 2 ) is discussed.     

Reduktion von 1,2-bis[(Z)-(2-nitrophenyl)-NNO-azoxy]benzol: Synthese von Cyclotrisazobenzol ( = (5E,6aZ,11E,12aZ,17E,18aZ)-5,6,11,12,17,18-Hexaazatribenzo[aei][1,3,5,7,9,11]cyclododeca-hexaen)

Reduction of 1,2-Bis[(Z)-(2-nitrophenyl)-NNO-azoxy]benzene¹: Synthesis of Cyclotrisazobenzene ( = (5E,6aZ,11E,12aZ,17E,18aZ)-5,6,11,12,17,18-Hexaazatribenzo[aei][1,3,5,7,9,11]cyclododeca-hexaene) Na₂S reduction of 1,2-bis[(Z)-(2-nitrophenyl)-NNO-azoxy]benzene ( 2 ) yielded 3 deoxygenated products: the (known) red 2,2′-((E,E)-1,2-phenylenbisazo)dianiline ( 3 , 23%), the orange 2-[2-((E)-2-aminophenylazo)phenyl]-2H-benzotriazol ( 4 , 55%) and the colorless 2,2′-(1,2-phenylene)di-2H-benzotriazol ( 5 , 13%). The constitutions of 3 – 5 and of 6 , the N-acetyl derivative of 4 , were deduced from their ¹H-NMR spectra (chemical shifts, couplings, and symmetry properties), and the configurations of 3 , 4 , and 6 at their N,N-double bonds are assumed to be the same as in 2 . Oxidation of 3 with 2 mol-equiv. of Pb(OAc)₄ afforded 5 (47%) and a novel, highly symmetrical macrocycle, called cyclotrisazobenzene ( 7 , 24%). The constitution of 7 as a tribenzo-hexaaza[12]annulene and its (E)-configuration at the N,N-bonds was confirmed by X-ray analysis. The molecular symmetry expressed by the ¹H-, ¹³C- and ¹⁵N-NMR spectra of 7 reveals a rapid torsional motion around the six N,C bonds. This implies that the N,N-double bonds in the cyclic 12π-electron system (or 24π-electron system if the benzene rings are included) of 7 are highly localized.     

A study on the sulfenylchlorination of cembrene and the related cembranoids

p-Tolylsulfenyl chloride adds regioselectively at the C(11) ? C(12) bond of cembrene 1 to give 11-(R)-p-tolylthio-1-(S)-cembra-2E, 4Z, 7E, 12(20)-tetraene 2 following a rapid HCl elimination. Two new cembranoids, 1-(S)-cembra-2E, 4Z, 7E, 12(20)-tetraene 3 and 1-(S)-cembra-2E, 4Z, 7E, 11Z-tetraen-20-ol 4 are prepared from 2.     

Secoiridoid Glycosides from Swertia mileensis

From the MeOH extract of the aerial parts of Swertia mileensis, four new secoiridoid glycosides were isolated, 4′‐O‐[(E)‐caffeoyl]swertiamarin ( 1 ), 4′‐O‐[(Z)‐coumaroyl]swertiamarin ( 7 ), 6′‐O‐[(E)‐coumaroyl]swertiamarin ( 8 ), and 6′‐O‐[(Z)‐coumaroyl]swertiamarin ( 9 ), together with five known compounds. Their structures were elucidated by NMR spectroscopy and tandem mass spectrometry. Detailed HPLC/MS analyses and MS/MS fragmentation pathways are discussed for the identification of the swertiamarin‐derived (E)/(Z) isomers 6 / 7 and 8 / 9 .     

Specifically (π → π*)-Induced Cyclohexenone Reactions 4a-(Z-1-Propenyl)-bicyclo[4.4.0]dec-1 (8a)-en-2-one and 4a-Z-1-Propenyl-bicyclo[4.4.0]deca-1(8a), 7-dien-2-one

4a-(Z-1-Propenyl)-bicyclo[4.4.0]dec-1(8a)-en-2-one ( 6 ) and 4a-(Z-1-propenyl)-bicyclo[4.4.0]deca-1(8a), 7-dien-2-one ( 17 ) undergo an intramolecular hydrogen transfer from the methyl group of the propenyl substitutent to the α-carbon atom of the enone group, and cyclization to the [4.4.3]propellane derivatives 9 and 30 , respectively, when excited in the π → π* wavelength region. The quantum yield for (Z)- 6 → 9 under optimum conditions is 0.29 at 254 nm. These reactions occur specifically from the S₂ (π,π*) state, competing with the S₂ → T decay. The triplet reactions of 6 are E–Z double-bond isomerization, double-bond shift to (E,Z)- 8 , and rearrangement to (E)- 10 . Further investigations concern some structural limitations in the scope of the reaction type 6 → 9 and enone S₂ reactivity in general.     

Polar Metabolites of the Tropical Green Seaweed Caulerpa taxifolia Which Is Spreading in the Mediterranean Sea: Glycoglycerolipids and Stable Enols (α\=Keto Esters)

Examination of the polar components of the green seaweed Caulerpa taxifolia (Vahl ) C. Agardh , which is heavily spreading in the northeastern Mediterranean, led to two families of compounds. The new (2R)-3-O-β-D -galactopyranosyl-1-O-[(7Z,10Z,13Z)-hexadeca-7,10,13-trienoyl]-2-O-[(9Z,12Z,15Z)-octadeca-1,12,15-trienoyl]-sn-glycerol ( 2 ) was isolated in low abundance, like the analogues 1 and 3 already known from freshwater cyanobacteria. The acyl positions in 1 – 3 were determined by enzymatic methods and the absolute configuration from the O-galactosylglycerol obtained upon alcaline methanolysis. More abundant were the (4-hydroxyphenyl)- and (3,4-dihydroxyphenyl)pyruvic acid methyl esters, occurring in the enol (Z) forms 13a and 14a accompanied by very minor (E) forms 13b and 14b . The latter became predominant on UV irradiation of 13a or 14a , allowing the determination of the C=C configuration of these isolatable, stable enols from ¹H,¹³C NMR couplings (larger H−C(3)/C(1) coupling constant in the (E) than in the (Z) isomer). Contrary to literature implications, the O-galactosylglycerolipids 1 – 3 lack any cholinergic or histaminergic activity; similarly, enols (= α-keto esters) 13 and 14 or terpenoids of this seaweed were also devoid of such biological activities (see Table).     

选择性响应Cu~(2+)、Ag~+及阴离子的菲咯啉-水杨醛荧光探针

在乙酸酐中用2,9-二甲基-1,10-菲咯啉与水杨醛缩合反应得到2,2′-(1E,1′E)-2,2′-(1,10-菲咯啉-2,9-二基)双(乙烯-2,1-二基)双(2,1-亚苯基)二乙酸酯(探针1);再将其进一步水解得到2,2′-(1E,1′E)-2,2′-(1,10-菲咯啉-2,9-二基)双(乙烯-2,1-二基)二苯酚(探针2)。经1H NMR、13C NMR、IR、MS表征,探针化合物为大共轭结构,发光性能良好。两种探针分别表现出对Cu2+、Ag+不同的荧光猝灭作用,探针2还能识别阴离子F-和AcO-,具有双功能离子检测性能。光谱滴定、等温滴定量热及质谱等测定了配合物组成、作用常数及热力学参数,探针与金属离子的配合为放热反应,作用比为2∶1。