JPH06321950A - Water-soluble lankacidin derivative - Google Patents

Abstract

PURPOSE:To obtain a new compound belonging to antibiotic of lankacidin group and useful as a therapeutic agent for infectious diseases. CONSTITUTION:A compound of the formula (R1 and R2 are OH, lower alkanoyloxy or acyloxy derived from amino acid; R3 is OH or lower alkanoyloxy and R'3 is H or R3 and R'3 together form oxo; at least one of R1 to R3 is acyloxy derived from optically active amino acid. This compound of the formula is obtained by subjecting a lankacidin antibiotic having a reactive hydroxyl group to ester condensation reaction with a reagent having a reactive carboxyl group in a solvent (e.g. DMF) at -20 to 80 deg.C. The compound of the formula exhibits strong antimicrobial action and is useful as an antibacterial agent against MRSA, etc., and its dose is 0.1-5g/adult.day.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a novel compound useful as a therapeutic agent for infectious diseases caused by pathogenic microorganisms. More specifically, the present invention relates to a novel compound belonging to the Lankacidin group antibiotics which is useful as a therapeutic agent for infectious diseases.

[0002]

Or [Prior Art] lankacidin group antibiotics produced by bacteria Sho 58-179496 publication typified by microorganisms eg Streptomyces rochei var. Volubilis, microbiological [JP from the product of the fungus 59-18
3695 and 63-245683] or a 17-membered ring antibiotic represented by the following general formulas [II] and [III] produced by conversion by a chemical method and JP-A-58-52285. The compound is also a generic name and is also referred to as antibiotic T-2636. Manufacturing method, physical properties, structural formula of these antibiotics and ester derivatives,
Regarding the fate and biological activity in the body, Chemical Pharmaceutical Bulletin (Chem. Pharm.Bull.),
Vol. 22, pp. 99-108 (1974) and The Journal of Antibiotics (J. Antibio
tics), 26, 647-657 and 658-6.
68 (1973) and the like.

[0003]

[Chemical 2]

[0004]

[Chemical 3] Further, as a result of the subsequent research, regarding the compounds represented by the formulas [IV] and [V], JP-A-58-52285, Japanese Patent Application Nos. 03-293475 and 04-18847.
No. 4 patent application specification and Chemical Pharmaceutical Bulletin (Chem. Pharm. Bull.), Volume 23,
2201-2210 (1975), etc., their chemical properties are described.

[0005]

[Chemical 4]

[0006]

[Chemical 5] Further, USP 4,914,206 describes the chemical properties of ester derivatives of lancasidin antibiotics.

[0007]

Diseases caused by bacteria have been largely overcome by the development of therapeutic regimens by antibiotic administration. However, the change of the causative bacteria (bacterial replacement phenomenon) or the emergence of resistant bacteria (resistance phenomenon) due to long-term or large-scale administration of conventional antibiotics has become a major problem in the current medical field of infectious diseases. In particular, MRSA (Methicillin-resistant St
Aphylococcus aureus ) has few effective antibiotics, which is a social problem. To overcome these problems, there is a constant need in the art for new types of infectious disease therapeutic agents.

[0008]

Means for Solving the Problems In view of the present situation, the present inventors have conducted research from a new viewpoint, and have obtained the following findings. That is, the Lancacidin group antibiotics have a unique structure that forms a ring with 17 carbons, do not show cross resistance to clinical isolates of Staphylococcus aureus, and are effective in in vivo tests. Was recognized. However, these antibiotics are fat-soluble, and only attempts to develop them as oral antibacterial agents were considered. Therefore, the present inventors have come to consider developing a water-soluble derivative of lancasidin C, which exhibits the strongest activity among antibiotics of this strain, as an anti-MRSA injection. When various derivatives were examined for water solubility, stability, antibacterial property, toxicity, etc., it was found that a water-soluble ester derivative, particularly a compound obtained by condensing an amino acid with lancasidin C or lancasidinol, etc. was the most useful.

The present inventors have conducted further research based on these findings and completed the present invention. That is, the present invention has the general formula

[Chemical 6] [Wherein R ₁ and R ₂ are a hydroxyl group, a lower alkanoyloxy group or an acyloxy group derived from an amino acid, R ₃ is a hydroxyl group, a lower alkanoyloxy group or an acyloxy group derived from an amino acid and R ₃ ′ is hydrogen] , R
₃ and R ₃ 'represent an oxo group, and at least one of R ₁ , R ₂ and R ₃ is an acyloxy group derived from an optically active amino acid] or a salt thereof and a compound [I] It provides an antibacterial agent.

In the formula, the alkanoyl group of the lower alkanoyloxy group represented by R ₁ , R ₂ and R _{3 includes} , for example, formyl, acetyl, propionyl, butyryl, isobutyryl, valeryl, isovaleryl, pivaloyl, hexanoyl and the like. , C _1-3 alkanoyloxy is preferred. The amino acid-derived acyloxy group means a state in which a carboxylic acid of an amino acid is condensed with a hydroxyl group. In this specification, when an abbreviation is used for an amino acid or the like, the IUPAC-IUB commission
On Biochemical No Men Creature (Commis
sion on Biochemical Nomenclature) or an abbreviation commonly used in this field, and
When an amino acid may have an optical isomer, the L form is shown unless otherwise specified. Regarding the above-mentioned general formula [I], the amino acid of the amino acid residue is supervised by Mitsuo Kotake; Dai Organic Chemistry Vol. 21, Natural Polymer Compound III, Asakura Shoten, 19
60 years and edited by JSDavies; amino acids and peptides (Amin
o acids and peptides), Chapman and Hall, 1985
As described in the years, etc., it means a compound having an amino group and an acidic group in a molecule which may have a substituent, and a usual amino acid constituting a protein, for example, an aliphatic monoaminomonocarboxylic acid. Acids; glycine (Gly), alanine (Ala), valine (Val), leucine (Leu), isoleucine (Ile), aliphatic oxyamino acids; serine (Se)
r), threonine (Thr), acidic amino acids and acidic amino acid amides; aspartic acid (Asp), asparagine (A
sn), glutamic acid (Glu), glutamine (Gln), aromatic amino acids; phenylalanine (Phe), tyrosine (Ty)
r), tryptophan (Try), iminocarboxylic acid; proline (Pro), oxyproline, basic amino acids; arginine (Arg), lysine (Lys), histidine (His),
Sulfur-containing amino acids; methionine (Met), cystine (Cys),
Examples include cysteine.

Abnormal amino acids obtained from the natural world as microbial metabolites or animal and plant components, for example, aliphatic monoaminomonocarboxylic acid; L-α-aminobutyric acid, γ-aminobutyric acid, β-aminoisobutyric acid, β-alanine, homoserine. , Α-methyl-D-serine, 0-carbamyl-D-serine, δ-hydroxy-γ-oxo-nor-valine (HON), monoaminodicarboxylic acid; L-α-aminoadipic acid, L-theanine, L -Γ-methylene glutamic acid, L-γ-methyl glutamic acid, 3-aminoadipic acid, aziridine 2,3-dicarboxylic acid (ADA), diaminomonocarboxylic acid; L-ornithine, β-lysine,
α, β-diaminopropionic acid, L-α, γ-diaminobutyric acid, diaminodicarboxylic acid; diaminopimelic acid, acidic amino acid; cysteic acid, taurine, aromatic amino acid; kynurenine, 3,4-dioxyphenyl-L-alanine, Heterocyclic amino acids; [S] -2-amino-3- (isoxazolin-5-on-4-yl) -propionic acid,
Anticapsin, basic amino acid; L-4-oxalidine, L-4-oxolysine, [3R, 5R] -3,6-
Diamino-5-hydroxyhexanoic acid, sulfur-containing amino acid; lanthionine, S-methyl-L-cysteine, cyclic amino acid; pipecolic acid, azetidine-2-carboxylic acid, [1R, 2S] -2-aminocyclopentane-1-
Carboxylic acid (ACPC), special functional group-substituted amino acids; citrulline, alanosine, L-azaserine, amino acids having various substituents obtained by an organic synthesis method, and the like can be mentioned. When these amino acid residues may have optical isomers, they may have either L- or D-configuration.

The amino acids constituting the acyloxy group derived from the above amino acids are preferably lysine, alanine, leucine, proline, aziridine, 2,3-dicarboxylic acid and 2-aminocyclopentane-1-carboxylic acid, and particularly optical. Active D-alanine, [1R, 2S]
It is preferably 2-aminocyclopentane-1-carboxylic acid. Further, as the compound [I], preferred is one in which R ₁ is an acyloxy group of the above-mentioned optically active amino acid, R ₂ is a hydroxyl group, and R ₃ and R ₃ ′ are oxo groups. Further, the compound [I] is preferably a water-soluble compound having a solubility in water of 30 mg / ml or more, and is preferably a compound stable in an aqueous solution of pH 4-5.

Next, a method for producing the above compound [I] will be described. The condensate, which is a raw material compound used in the production of the general formula [I] or a salt thereof, comprises a reagent having a reactive carboxyl group and a lancacidin antibiotic having the other reactive hydroxyl group by using a conventional ester synthesis method. It can be produced by condensation. Examples of the compound having activated carboxyl group as a raw material include active ester [eg, alcohol (eg, pentachlorophenol, 2,4,5-
Trichlorophenol, 2,4-dinitrophenol,
Cyanomethyl alcohol, p-nitrophenol, N,
N'-dicyclohexylcarbodiimide (DCC),
N, N'-diisopropylcarbodiimide, N-hydroxy-5-norbornene-2,3-dicarboximide, N-hydroxysuccinimide, N-hydroxyphthalimide, 1-hydroxybenztriazole (H
OBt), an ester with hydrochloric acid 1-ethyl-3- (3-dimethylaminopropyl) -carbodiimide (WSC), etc.] and the like. The condensation reaction can be carried out in the presence of a solvent. The solvent can be appropriately selected from those known to be usable in the ester condensation reaction.
For example, anhydrous formamide, dimethylformamide (DM
F), amides such as N-methylpyrrolidone, sulfoxides such as dimethyl sulfoxide (DMSO), amines such as pyridine and triethylamine (TEA),
Chloroform, dichloromethane (DCM) and other halogenated hydrocarbons, tetrahydrofuran (THF), dioxane and other ethers, acetonitrile and other nitriles, ethyl acetate (AcOEt), ethyl formate and other esters, or appropriate proportions thereof. And the like. The reaction temperature is appropriately selected from the range known to be applicable to ester bond formation reactions. Specifically, for example, it is usually appropriately selected from the range of about -20 ° C to 80 ° C. The reaction time is appropriately selected from the range known to be necessary for the ester bond formation reaction. Specifically, the reaction is performed for, for example, several minutes to 48 hours.

The water-soluble functional group of the amino acid used in the above condensation reaction is preferably protected. The protecting group in this case is a known group for protecting an amino group, a carboxyl group or a hydroxyl group in peptide synthesis, and is, for example, a protecting group eliminated by hydrolysis, hydrogenolysis, reduction, aminolysis or hydrazinolysis. Is. Amino group-protecting groups include groups derived from carbonic acid, such as benzyloxycarbonyl (Z), t
-Butyloxycarbonyl (Boc), t-amyloxycarbonyl, isobornyloxycarbonyl, 4-methoxybenzyloxycarbonyl, 2-chlorobenzyloxycarbonyl, adamantyloxycarbonyl, trifluoroacetyl, formyl, 9-fluorenylmethyl Examples include oxycarbonyl (Fmoc) and the like, nitrated arylsulfenyl groups (eg, 2-nitrophenylsulfenyl, etc.), arylphosphinothionyl groups (eg, diphenylphosphinothionyl, etc.), phthalyl and the like. Of these, groups derived from carbonic acid are particularly preferred.

The deprotection reaction of the above protecting group is carried out by a method known per se, for example, a method commonly used in peptide chemistry. Deacylation of the acyloxy group is carried out by hydrolysis with a haloacetic acid, for example, trifluoroacetic acid (TFA), or a hydrohalic acid, for example, hydrochloric acid, hydrobromic acid, etc., in a suitable solvent. be able to. The oxycarbonyl group is easily hydrogenolyzed using an organic base such as diethylamine, piperidine (PIP), morpholine, p-dimethylaminopyridine, dicyclohexylamine. Since the lancacidin group antibiotics are easily decomposed in an acidic aqueous solution, a reaction carried out in an organic solvent using an organic base is advantageous. Further, the 14-position acyl group and the 8-position acyl group can be eliminated by an enzymatic reaction. That is, the former Streptomyces rochei var. Vo
lubilis [Harada et al .; Journal of Antibiotics, 26, 647-65.
7 (1973)], the latter is Bacillus megaterium
IFO 12108 [Nakahama et al .; Journal of Antibiotics, 28, 3
90-394 (1975)] is hydrolyzed by esterase I and esterase II contained in the fermentation broth. By appropriately combining these selective enzyme reactions and deprotection reactions, water-soluble ester derivatives of lancasidin antibiotics can be easily prepared. The general method for collecting the desired lancasidin derivative from the reaction solution is described below. When the compound is neutral and lipophilic,
A general means that utilizes this property may be used. An organic solvent immiscible with water, such as chloroform, ethyl acetate, methyl isobutyl ketone, or isobutanol, is added to the obtained reaction solution to extract the lancacidin derivative. The extract is washed with aqueous sodium hydrogen carbonate and water, and the organic solvent layer is concentrated to give a crude substance.

Various chromatographic methods are advantageously used to further purify the crude material to obtain the pure compound. As the carrier, silica gel, crystalline cellulose, Sephadex LH-20 (Pharmacia, Sweden) and the like are used, and these are usually carried out by a column chromatography method. To elute the active substance from the column, a suitable organic solvent such as n-hexane, chloroform, toluene, ethyl acetate, dichloroethane, acetone, methanol or the like may be used alone or as a mixed solvent. The recrystallization method is advantageously used to further purify the crude crystal to obtain a pure target product. As the recrystallization solvent, for example, diethyl ether, ethyl acetate, acetone, methanol, ethanol, etc. are advantageously used. It can also be purified by preparative high performance liquid chromatography (HPLC). As the carrier, octadecylsilane (ODS) type and silica gel type carriers are advantageously used. For example, in the case of ODS, a mixed solution of methanol or acetonitrile and a salt-containing aqueous solution is advantageously used as the mobile phase. The eluate containing the target substance is extracted with a suitable organic solvent that is immiscible with water, and the extract is concentrated,
The residue is crystallized or powdered from the above-mentioned suitable organic solvent to obtain the desired product. When the derivative is water-soluble, particularly when it is an amino acid ester, its aqueous solution in salt form is used as an adsorbent resin such as Amberlite XAD-I and II (Rohm and Haas, USA), Diaion HP-.
It is advantageous to perform chromatography using 20 and SP-207 (manufactured by Mitsubishi Kasei Co.) as a carrier. As an elution solvent, a mixed solution of methanol or acetonitrile and water or an aqueous solution containing an acid or a salt is advantageously used. Further, the above-mentioned purification method is appropriately added to obtain the desired product. The compound [I] of the present invention can be obtained as an acid addition salt, particularly a pharmacologically acceptable acid addition salt, by a method known per se, for example, an inorganic acid (eg, hydrochloric acid,
Examples thereof include salts of sulfuric acid, phosphoric acid) or organic acids (eg, acetic acid, propionic acid, citric acid, tartaric acid, malic acid, oxalic acid, methanesulfonic acid).

The physicochemical properties (solubility and stability in aqueous solution) of compound [I] are described below.

[Table 1]

[Table 2] * Numerical values indicate percent survival of the compound. Stability test condition: Compound was dissolved in a concentration of 1 mg / ml (20% glucose solution), and the residual rate at a temperature of 24 ° C and 37 ° C was set to H.
It quantified by PLC. Solubility test conditions: 100 mg / ml or 50 mg / ml (20
% Glucose solution), after dissolving and suspending the compound, 0.2
It was filtered with a 2 μm filter, diluted, and quantified by HPLC. Measurement method; converted from HPLC peak area Measurement conditions Column: ODS YMC Pack A-312 Mobile phase: 60% MeOH / 0.02M phosphate buffe
r pH 7.5 flow rate; 1.0 ml / min detection method; 230 nm (UV) Next, the biological activity of these antibiotics will be described. [Table 3] shows the antibacterial activity of Lancacidin C8-ACPC (Compound 5 ) and Lancacidin C8-D-Ala (Compound 20 ) against various pathogenic bacteria.

[0018]

[Table 3] * 1 Bact Antibiotic Medium 3 (Difco Laboratories, USA) 17.5g, Bact
Yeast extract (Difco Laboratories, USA) 5.0 g, distilled water 1000 ml (no pH adjustment)
About 10 ⁶ CFU / m as inoculum
It was measured by the agar dilution method using l. Moreover, in [Table 2], MR of lancassidin C 8-ACPC (compound 5 ) and lancassidin C 8-D-Ala (compound 20 ) is shown.
The antibacterial activity against SA is shown.

[0019]

[Table 4] * 1 Minimum growth inhibitory concentration (10 ⁴ CFU by agar dilution method)
/ Spot) * 2 MH; Mueller Hinton medium, S-MH; Serum-containing Mueller Hinton medium Lancacidin C 8-ACPC (Compound 5 ) MRSA
When it was intravenously administered to (N133A strain) infected mice (n = 5), the ED ₅₀ value was 1.4 mg / kg, which showed a strong therapeutic effect. When the same compound was intravenously injected into a mouse and subjected to an acute toxicity test, no death was observed even at a dose of 100 mg / kg. As is clear from these data, the lancasidin derivative represented by the formula [I] exhibits antibacterial activity against many Gram-positive bacteria such as S. aureus containing multidrug-resistant bacteria and has no toxicity to mammals. Extremely weak. Therefore, the antibiotics can be widely used for treating infectious diseases caused by mammalian pathogenic microorganisms.

The antibacterial agent containing the compound of the present invention as an active ingredient is useful as a prophylactic and therapeutic agent for infections caused by mammalian pathogenic microorganisms, and is obtained by mixing the compound with a pharmacologically acceptable carrier. To be This preparation is a parenteral preparation such as injection, drip, liquid, suspension and suppository, and oral preparation such as capsule, tablet,
Syrups, powders and granules or other pharmaceutically suitable dosage forms can be provided. Parenteral agents, for example, isotonic agents (eg, glucose, sorbitol, mannitol, sodium chloride, etc.) when producing injections, preservatives (eg, benzyl alcohol, chlorobutanol, methyl parahydroxybenzoate, etc.), Anticoagulants (eg, dextran sulfate, heparin, etc.), solubilizing agents (eg, cyclodextrins, tween, etc.), stabilizers (eg, polyethylene glycol, polylactic acid, etc.) and the like may be contained. Upon administration, these antibiotics are dissolved in a conventional aqueous diluent and used as a solution. Diluents include dextrose aqueous solution, physiological saline, Ringer's solution, nutritional supplement solution, etc. Further, the oral preparation may contain additives such as an excipient, a binder, a disintegrant, a lubricant, a coloring agent, a corrigent, a stabilizer and the like. These preparations are orally or parenterally administered, and when used in humans, the dose may vary depending on the type and degree of the target disease, the age of the patient, etc. It is preferred that about 0.1 g to 5 g, and especially 0.2 g to 2 g, be used per person for the treatment of the disease.

[0021]

The contents of the present invention will be described in more detail with reference to the following examples, which should not be construed as limiting the present invention. The following abbreviations were used in Reference Examples and Examples. ACPC: [1R, 2S] -2-aminocyclopentane-1-carboxylic acid ADA: aziridine 2,3-dicarboxylic acid Fmoc: 9-fluorenylmethyloxycarbonyl TEA: triethylamine DCM: dichloromethane AcOEt: ethyl acetate PIP: piperidine WSC: hydrochloric acid 1-ethyl-3- (3-dimethylaminopropyl) -carbodiimide HOBt: 1-hydroxybenztriazole HOSu: N-hydroxysuccinimide

Reference Example 1 Amino acid, ACPC (3.0 g) was added to dioxane (60 m
l), dissolved in a mixture of water (60 ml), TEA (4.7 m
l), 9-Fluorenylmethylsuccinimidyl carbonate (Fmoc-OSu, 9.0 g) was added, and the mixture was stirred at room temperature for 3 hours. 2% sodium hydrogen carbonate water (300 m
l) was added, washed with diethyl ether (50 ml × 2), adjusted to pH 3.0 with 3N hydrochloric acid, and AcOEt (200
It was extracted with ml × 3). The ethyl acetate layer was dried over anhydrous sodium sulfate and then concentrated, and N-Fmoc-ACPC (10.8
g) was obtained as a colorless oil. Next, N-Fmoc-ACP
C (10.8 g) is dissolved in DCM (270 ml) and WS
C ^R (7.7g), was stirred for 3.5 hours at room temperature was added anhydrous HOBt (5.4 g). Hexane (250 m
l) and AcOEt (250 ml) were added, and saturated saline solution (10
After washing with 0x2), the organic layer was dried over anhydrous sodium sulfate and concentrated to give N-Fmoc-ACPC-OBu active ester (12 g) as a white powder.

Example 1 Lancacidin C 8- [1R,
2S] -2-Aminocyclopentanecarboxylate ( 5 ) Lancasidine A ( 1 , 7.5 g) in DCM (350 ml)
, N-Fmoc-ACPC-OBu active ester (11 g) and TEA (3.7 ml) were added, and the mixture was added at room temperature to 64
Stir for hours. Hexane (350 ml), AcO was added to the reaction mixture.
Et (350 ml) was added, and 10% aqueous ammonium chloride solution (250 ml) and 2% aqueous sodium hydrogen carbonate solution (200 ml x)
2) and washed with water (200 ml), respectively. The organic layer was dried over anhydrous sodium sulfate and then concentrated to dryness to obtain a crude substance (1
8.2 g) was obtained. This was subjected to silica gel chromatography (500 ml, Kieselgel 60, 70-230 mesh, manufactured by A Merck, Germany), developed with an eluent in which acetone was sequentially added in toluene, and Lancacidin was extracted from the 10% acetone elution fraction. A 8-N-Fmoc-ACPC
Was obtained as a white powder ( 4 , 8.7 g). Anal. Calcd. For C ₄₈ H ₅₄ N ₂ O ₁₁・ 0.25H ₂ O, C; 68.68, H; 6.54, N; 3.33 Found C; 68.69, H; 6.55, N; 3.34 UV λmax nm (ε); 207 (75,500), 220 (52,100), 226 (58,
800), 250 (35,100), 262 (25,900), 287 (7,000), 298 (7,8
00) (MeOH) IR νmax (cm ^-1 ); 1730, 1710, 1690, 1240 (KBr) Compound 4 (2.0 g, 2.4 mmol) in DCM (90 ml)
, And cooled to 0 ° C. PIP (10 ml) was added thereto, and the mixture was stirred at 0 ° C for 4 hours. After that, all the operations except the concentration operation were performed in a low temperature room (4 ° C.). AcOEt (5
00 ml) was added, and the mixture was washed with a solution prepared by diluting 1N hydrochloric acid water (98 ml) to 500 ml, and then 1N hydrochloric acid water (2.4 ml) was added to 20 ml.
Extract twice with a solution diluted to 0 ml, and the aqueous layer is immediately pH adjusted.
Adjusted to 5.0. The aqueous layer was washed several times with a mixed solution of diethyl ether and hexane (1: 1), adjusted to pH 5.0 again, and the organic solvent was distilled off with an evaporator using a vacuum pump (340 ml). Esterase I (70
0 mg) was added and the mixture was stirred at 4 ° C. for 16 hours. Acetone (1.4 liter) was added to the reaction solution, and the mixture was centrifuged (8.00 liters).
(0 rpm, 13 minutes), the supernatant was concentrated with a vacuum evaporator, and then adjusted to pH 4.0. Next, Amber Light XA
D-II (100 ml), washed with water (400 ml), then 20
Elution was performed with% acetonitrile water (500 ml). The eluate is concentrated and then lyophilized to give Lancascidin C 8-ACPC.
The hydrochloride salt (5, 820 mg) was obtained as a white powder. [Α] _D −190 ° (c 0.99, H ₂ O, 28 ° C.) Anal. Calcd. For C ₃₁ H ₄₂ N ₂ O ₈ · HCl · 2.5H ₂ OC; 57.09, H; 7.42, N; 4.30 , Cl; 5.44 Found C; 57.32, H; 7.34, N; 4.50, Cl; 5.29 UV λmax nm (ε); 226 (49,700) (MeOH) IR νmax (cm ^-1 ); 3400, 1710, 1680, 1510, 1380, 136
0, 1270, 1220, 1000,970 (KBr) ¹³ C NMR δ ppm; 75MHz, DMSO-d ₆ 210.1 (s), 196.4 (s), 171.1 (s), 170.2 (s), 159.6 (s),
137.1 (s), 136.7 (s), 135.5 (d), 135.5 (d), 131.0 (d), 1
26.1 (d), 125.7 (d), 125.4 (d), 76.3 (d), 75.0 (d), 67.8
(d), 56.0 (s), 52.1 (d), 51.2 (d), 45.8 (d), 45.7 (d),
37.2 (t), 33.0 (t), 29.8 (t), 26.6 (t), 24.4 (q), 21.0
(t), 20.2 (q), 12.3 (q), 12.2 (q), 9.1 (q)

Example 2 Lancacidin A 8- [1R,
2S] -2-Aminocyclopentanecarboxylate ( 6 ) Compound 4 (1.80 g) obtained in Example 1 was added to DCM.
It was dissolved in (27 ml) and ice-cooled to 0 ° C. PIP (3.0
ml) was added and the mixture was stirred at 0 ° C. for 4 hours. Ether (150
ml), washed with water (100 ml × 2) and 0.015N hydrochloric acid water (100 ml), and then washed with 0.15N hydrochloric acid water (100 ml).
Immediately after extracting the desired product with (ml), the pH of the aqueous layer was adjusted to 5.5. Here, after combining 3 times, hexane (50 ml x
After washing with 3), add AcOEt (150 ml x
Extracted in 3). The AcOEt layer is 0.25N hydrochloric acid water (13
After extracting with 0 ml × 5) and immediately adjusting the pH to 3.0,
After removing AcOEt with an evaporator, Amberlite X
After applying to AD-II (65 ml) and washing with water (195 ml), 20
Elution was performed with% methanol water (300 ml), 50% methanol water (300 ml), 80% methanol water (300 ml) and methanol (300 ml). The eluate was concentrated and lyophilized, lankacidin A 8-ACPC hydrochloride (6, 5
26 mg) was obtained as a white powder. [Α] _D −120 ° (c 0.51, H ₂ O, 21 ° C.) Anal. Calcd. For C ₃₃ H ₄₄ N ₂ O ₉ · HCl · 2H ₂ OC; 57.85, H; 7.21, N; 4.09, Cl; 5.17 Found C; 57.93, H; 7.08, N; 4.17, Cl; 5.11 UV λmax nm (ε); 226 (41,600) (MeOH) IR νmax (cm ^-1 ); 3410, 1730, 1710, 1690, 1510 , 1250
(KBr) ¹³ C NMR δ ppm (75 MHz, DMSO-d ₆ ) 210.4 (s), 196.4 (s), 171.0
(S), 170.1 (s), 169.4 (s),
159.6 (s), 140.2 (d), 137.2
(S), 136.1 (s), 135.4 (d),
128.0 (d), 126.2 (d), 125.6
(D), 124.6 (d), 76.1 (d), 7
4.4 (d), 71.2 (d), 56.0 (s),
52.1 (d), 51.3 (d), 45.8
(D), 45.7 (d), 33.7 (t), 33.
1 (t), 29.8 (t), 26.5 (t), 2
4.4 (q), 21.0 (q), 21.0 (t),
20.3 (q), 12.2 (q), 12.0
(Q), 8.9 (q)

Example 3 Lancacidin C 8,14-di [1R, 2S] -2-aminocyclopentanecarboxylate ( 7 ) Lancacidin C ( 2 , 3.37 g) was added to DCM (230 m).
l-), N-Fmoc-ACP obtained in Reference Example 1
C-OBt active ester (6.90 g), TEA (2.
45 ml) was added and the mixture was stirred at room temperature for 72 hours. Furthermore N
-Fmoc-ACPC-OBt active ester (7.00
g) and TEA (2.45 ml) were added, and the mixture was stirred at room temperature for 88 hours. Hexane (200 ml), AcOEt (200 m
l) was added and 10% aqueous ammonium chloride (200 ml),
Sodium hydrogencarbonate water (200 ml x 2), water (200 m
After washing with l), drying over anhydrous sodium sulfate, and concentrating,
Crude material (13.1 g) was obtained. This was chromatographed on silica gel (500 ml), developed with an eluent in which acetone was sequentially added to toluene, and 10% acetone eluted fractions were used to extract lancascidin C 8,14-di-N-Fmoc.
-ACPC ( 7 , 5.14g) was obtained as a white powder. Anal. Calcd. For C ₆₇ H ₇₁ N ₃ O ₁₃・ 0.5C ₇ H ₈ C; 72.22, H; 6.45, N; 3.58 Found C; 72.23, H; 6.56, N; 3.62 UV λmax nm (ε); 207 (173,000), 220 (173,000), 226 (1
44,000), 254 (110,000), 262 (102,000), 228 (24,800), 2
98 (28,200) (MeOH) IR νmax (cm ^-1 ); 1720, 1690, 1510 (KBr) Compound 7 (1.0 g) was dissolved in DCM (45 ml) and 0
After ice cooling to ℃, PIP (5.0 ml) was added, and the mixture was stirred at 0 ℃ for 4 hours. AcOEt (200 ml) was added, and water (200 ml) was added.
X2), after washing with 0.013N hydrochloric acid water (200 ml),
It was extracted with 0.13N hydrochloric acid water (200 ml). This was immediately adjusted to pH 4.0, washed with ethyl acetate (100 ml), and then the organic solvent was distilled off with an evaporator using a vacuum pump. This was chromatographed on Diaion HP-20 (50 ml), washed with water (200 ml), and then 20
% Methanol water (200 ml), 50% methanol water (2
Eluted with 100 ml), After concentration and freeze drying, lankacidin C 8,14-di -ACPC2 hydrochloride (8, 370 mg)
Was obtained as a white powder. [Α] _D −108 ° (c 0.
50, H ₂ O, 21 ° C.) Anal. Calcd. For C ₃₇ H ₅₁ N ₃ O ₉・ 2HCl ・ 3H ₂ OC; 54.95, H; 7.35, N; 5.20, Cl; 8.77 Found C; 55.02, H; 7.32 , N; 5.29, Cl; 8.47 UV λmax nm (ε); 226 (46,100) (MeOH) IR νmax (cm ^-1 ); 3400, 1710, 1630, 1510, 1460, 1220
(KBr) ¹³ C NMR δ ppm (75MHz, DMSO-d ₆ ) 210.6 (s), 196.4 (s), 171.0 (s), 170.8 (s), 170.1 (s),
159.6 (s), 140.3 (d), 137.1 (s), 136.2 (s), 135.3 (d), 1
28.1 (d), 126.2 (d), 125.7 (d), 124.5 (d), 76.1 (d), 74.
6 (d), 72.2 (d), 56.0 (s), 52.1 (d), 52.1 (d), 51.3 (d),
45.8 (d), 45.7 (d), 45.7 (d), 33.5 (t), 33.1 (t), 29.8
(t), 29.7 (t), 26.5 (t), 26.4 (t), 24.4 (q), 21.0 (t), 2
1.0 (t), 20.3 (q), 12.2 (q), 12.0 (q), 9.0 (q)

Example 4 Lancasidinol 2 ', 8-
[1R, 2S] -2-Aminocyclopentanecarboxylate ( 9 ) Lancasidinol A ( 3 , 3.69 g) was added to DCM (23
0 ml), and N-prepared in Reference Example 1 in this solution.
Fmoc-ACPC-OBt active ester (7.0 g), T
EA (2.45 ml) was added, and the mixture was stirred at room temperature for 84 hours.
Furthermore, N-Fmoc-ACPC-OBt active ester (6.
3 g) and TEA (2.2 ml) were added, and the mixture was stirred at room temperature for 48 hours. Hexane (300 ml) and AcOEt in the reaction solution.
(300 ml) was added, and 10% ammonium chloride water (30
0 ml), 2% aqueous sodium hydrogen carbonate solution (300 ml x 2),
After washing with water (300 ml), dehydration with anhydrous sodium sulfate,
The organic solvent layer was concentrated to dryness to obtain a crude material (13.5 g). This is chromatographed on silica gel (800 m
l) and developed with an eluate in which acetone in toluene was sequentially added, and 10% acetone-eluted fraction was used to extract Lancasidinol A 2 ′, 8-di-Fmoc-ACPC ( 9 , 4.73).
g) was obtained as a white powder. Anal. Calcd. For C ₆₉ H ₇₅ N ₃ O ₁₄・ 0.5H ₂ OC; 70.27, H; 6.50, N; 3.56 Found C; 70.05, H; 6.54, N; 3.65 UV λmax nm (ε); 208 (81,500 ), 220 (42,100), 226 (35,
100), 252 (25,700), 262 (23,900), 288 (5,900), 298 (7,0
00) (MeOH) IR νmax (cm ^-1 ); 1730, 1710, 1510, 1240 (KBr) Compound 9 (2.0 g) was dissolved in DCM (45 ml), and 0
It was ice-cooled to ℃. PIP (5.0 ml) was added, and the mixture was stirred at 0 ° C for 4 hr. After adding AcOEt (200 ml), water (2
The mixture was washed with 00 ml × 2) and 0.013 N hydrochloric acid water (200 ml), the target product was extracted with 0.13 N hydrochloric acid water (200 ml), and the pH of the aqueous layer was immediately adjusted to 5.0. Here, the two portions were combined, washed with hexane (100 ml × 3), the aqueous layer was concentrated, and the organic solvent was evaporated. Esterase I (1.
2 g) was added, the mixture was stirred at room temperature for 1 hour, and then extracted with isobutyl alcohol (200 ml × 10). Next, 0.01N
It was re-extracted with hydrochloric acid water (500 ml × 2), adjusted to pH 5.0 and then concentrated, and the organic solvent was distilled off. This was subjected to HP-20 chromatography (100 ml) and water (300 m).
After washing with l), 20% methanol water (400 ml), 50
Elution was performed with% methanol water (400 ml). The eluate was concentrated and lyophilized to give lancasidinol 2 ', 8-di-A.
CPC2 hydrochloride ( 10 , 512 mg) was obtained as a white powder. [Α] _D −106 ° (c 0.52, H ₂ O, 21 ° C.) Anal. Calcd. For C ₃₇ H ₅₃ N ₃ O ₉・ 2HCl ・ 3H ₂ OC; 54.81, H; 7.58, N; 5.18, Cl; 8.75 Found C; 54.57, H; 7.65, N; 5.17, Cl; 8.61 UV λmax nm (ε); 226 (40,500) (MeOH) IR νmax (cm ^-1 ); 3410, 1730, 1710, 1660, 1520 , 138
0, 1270, 1210 (KBr) ¹³ C NMR δ ppm; 75MHz, DMSO-d ₆ 209.9 (s), 171.1 (s), 171.1 (s), 170.6 (s), 170.0 (s),
136.7 (s), 136.5 (s), 135.6 (d), 135.6 (d), 131.0 (d), 1
26.1 (d), 126.0 (d), 125.4 (d), 76.3 (d), 74.9 (d), 70.5
(d), 67.9 (d), 56.8 (s), 52.4 (d), 52.1 (d), 50.5 (d),
45.7 (d), 45.7 (d), 45.6 (d), 37.2 (t), 33.0 (t), 29.7
(t), 29.6 (t), 26.5 (t), 26.5 (t), 21.1 (t), 21.0 (t), 2
0.0 (q), 17.3 (q), 12.2 (q), 12.2 (q), 9.2 (q)

Example 5 Lancacidin C 14- [1
R, 2S] -2-Aminocyclopentanecarboxylate ( 12 ) Lancacidin C 8-butyrate (4.90 g) was added to DC.
Dissolved in M (150 ml), N-Fmo obtained in Reference Example 1
c-ACPC-OBt active ester (5.51 g), TE
A (1.96 ml) was added, and the mixture was stirred at room temperature for 84 hours. Add hexane (150 ml) and ethyl acetate (150 ml),
It was washed with 10% aqueous ammonium chloride (150 ml), 2% aqueous sodium hydrogen carbonate (200 ml × 2) and water (200 ml), dried over anhydrous sodium sulfate and concentrated to obtain a crude substance (10.3 g). . This was subjected to silica gel chromatography (500 ml), developed with an eluent in which acetone was sequentially added to toluene, and 5% to 10% acetone eluted fractions were used to analyze lancascidin C 8-butyryl-14.
-N-Fmoc-ACPC ( 11 , 5.46 g) was obtained as a white powder. Anal. Calcd. For C ₅₀ H ₅₈ N ₂ O ₁₁ C; 69.59, H; 6.77, N; 3.25 Found C; 69.39, H; 6.75, N; 3.49 UV λmax nm (ε); 208 (68,700), 220 ( 58,700), 226 (54,
900), 250 (31,900), 262 (24,200), 288 (5,500), 298 (6,0
00) (MeOH) IR νmax (cm ^-1 ); 1730, 1710, 1690, 1510, 1260 (KBr) Compound 11 (4.0 g) was divided into 2 portions and dissolved in methanol (1.0 liter). Add 0.1 M phosphorus buffer (9.0 liters, pH 8.0) and add esterase.
Wet cells containing II (250 g) were added, and then 48 at 37 ° C.
Stir for hours. After adding ethyl acetate (4.0 liter), the mixture was further stirred for 1 hour and filtered with Hyflo Supercel (trade name) (Jones & Manville Co., USA). 1.0 liter)
The extract was washed with water, dried over anhydrous sodium sulfate, and concentrated to obtain a crude substance (6.6 g). This was subjected to silica gel chromatography (300 ml) and developed with an eluent in which ethyl acetate was sequentially added to hexane, and the 60% ethyl acetate eluate fraction was used to analyze Lancascidin C 14 -N-Fmoc-ACPC.
(2.31 g) was obtained as a white powder. A part (800 mg) of this was dissolved in DCM (36 ml) and cooled to 0 ° C. PIP (4.0 ml) was added, and the mixture was stirred at 0 ° C for 4 hr. After adding ethyl acetate (200 ml), it was washed with 0.1 N hydrochloric acid water (392 ml), and 0.01 N hydrochloric acid water (100 ml).
It was extracted in × 2). Immediately after adjusting the pH of the water layer to 5.0,
After washing with a hexane-ether mixed solution (50 ml × 5), the mixture was concentrated and the organic solvent was distilled off. After adjusting this to pH 4.0, it was subjected to Amberlite XAD-II (40 ml),
After washing with water (200 ml), 20% acetonitrile water (2
(00 ml). After this concentration, was lyophilized, lankacidin C 14-ACPC hydrochloride (12, 440 mg)
Was obtained as a white powder. [Α] _D −114 ° (c 0.54, H ₂ O, 21 ° C.) Anal. Calcd. For C ₃₁ H ₄₂ N ₂ O ₈ · HCl · 2.5H ₂ OC; 57.09, H; 7.42, N; 4.30 , Cl; 5.44 Found C; 57.25, H; 7.49, N; 4.45, Cl; 5.85 UV λmax nm (ε); 226 (43,800) (MeOH) IR νmax (cm ^-1 ); 3400, 1710, 1680, 1510, 1360, 1260
(KBr) ¹³ C NMR δ ppm (75MHz, DMSO-d ₆ ) 210.6 (s), 196.5 (s), 170.9 (s), 170.1 (s), 159.5 (s),
140.7 (s), 137.6 (s), 134.6 (d), 132.5 (d), 132.3 (d), 1
30.3 (d), 124.0 (d), 123.8 (d), 74.5 (d), 72.7 (d), 72.4
(d), 56.2 (s), 52.0 (d), 51.2 (d), 45.8 (d), 45.6 (d),
37.3 (t), 33.6 (t), 29.9 (t), 26.6 (t), 24.4 (q), 21.0
(t), 20.3 (q), 12.3 (q), 11.9 (q), 9.0 (q)

Example 6 Lancacidin C 8- (4-oxa) -L-lysine ester ( 14 ) 4-oxa-L-Lys (9.21 g) in water (100 ml)
Dissolved in dioxane (100 ml), TEA (27.7)
ml), Fmoc-OSu (33.7 g) was added, and the mixture was stirred at room temperature for 3 hours. 10% ether / hexane solution (10
After washing with 0 ml × 2), the aqueous layer was concentrated and dioxane was distilled off. After adjusting the pH to 1.0 with hydrochloric acid, the mixture was extracted with ethyl acetate (200 ml × 2). The ethyl acetate layer was washed with water (100 ml × 3) and saturated saline (80 ml), dried over anhydrous sodium sulfate, concentrated, powdered from ether-hexane, and Nα, Nε-di-Fmoc-4-oxa-. L-Lys (18.1 g) was obtained as a white powder.
This powder (15.4 g) was dissolved in DCM (500 ml), WSC (5.5 g) and HOBt (3.9 g) were added, and the mixture was stirred at room temperature for 3 hours. Next, Lancacidin A (13
g) and TEA (4.0 ml) were added, and the mixture was stirred at room temperature for 48 hours. Hexane (500 ml) and ethyl acetate (5
(00 ml) was added, and then 10% ammonium chloride aqueous solution (400 ml) and 2% aqueous sodium hydrogen carbonate solution (400 ml x
2) and washed with water (400 ml), respectively. The organic layer was dried over anhydrous sodium sulfate, concentrated, subjected to silica gel chromatography (600 ml), developed with an eluent in which acetone in toluene was sequentially added, and the 5% to 10% acetone eluate fraction was concentrated, Powdered from ether-hexane to give Lancascidin A 8-Nα, Nε-di-Fmoc-4.
-Oxa-L-Lys ( 13 , 12.8 g) was obtained as a white powder. Anal. Calcd. For C ₆₂ H ₆₅ N ₃ O ₁₄・ 0.5H ₂ OC; 68.62, H; 6.13, N; 3.87 Found C; 68.78, H; 6.28, N; 3.78 UV λmax nm (ε); 208 (96,300 ), 220 (65,600), 226 (5
6,300), 250 (39,000), 262 (34,400), 288 (8,600), 298
(9,700) (MeOH) IR νmax (cm ⁻¹ ); 1730, 1710, 1510, 1250 (KBr) Compound 13 (6.0 g) was dissolved in DCM (270 ml), cooled to 0 ° C., and PIP (30 ml). And add 4 at 0 ℃
Stir for hours. Add ethyl acetate (1.2 liters),
0.59N hydrochloric acid water (500ml), 0.22N hydrochloric acid water (5
After extraction with 00 ml × 2), the pH was adjusted to 6.0, washed with ethyl acetate (200 ml × 3), adjusted to pH 5.0 and concentrated, and the organic solvent was distilled off (400 ml). Methanol (20 ml) and esterase I (1.0 g) were added thereto, and the mixture was stirred at room temperature for 2 hours. Acetone (1.6
Liter), centrifuge, concentrate the supernatant, and
It was adjusted to 4.0. It was then chromatographed on HP-20 (60 ml), eluting with water (240 ml). Reapply HP-20 (60 ml), wash with water (240 ml),
Eluted with 20% aqueous acetonitrile (240 ml), concentrated and lyophilized to give lancascidin C8- (4-oxa).
-L-Lys 2 hydrochloride (14, 620 mg) was obtained as a white powder. [Α] _D −143 ° (c 0.52, H ₂ O, 21 ° C.) Anal. Calcd. For C ₃₀ H ₄₃ N ₃ O ₉ · 2HCl · 2H ₂ OC; 51.58, H; 7.07, N; 6.01, Cl; 10.15 Found C; 51.48, H; 7.23, N; 6.01, Cl; 10.31 UV λmax nm (ε); 226 (47,500) (MeOH) IR νmax (cm ^-1 ); 3430, 1740, 1700, 1680, 1630 , 1510,
1260 (KBr) ¹³ C NMR δ ppm (75MHz, DMSO-d ₆ ) 211.0 (s), 196.5 (s), 170.3 (s), 166.5 (s), 159.7 (s),
137.0 (s), 136.9 (s), 136.0 (d), 135.5 (d), 131.3 (d), 1
26.2 (d), 125.3 (d), 125.0 (d), 77.5 (d), 75.0 (d), 67.9
(d), 67.6 (t), 66.8 (t), 56.1 (s), 52.4 (d), 51.2 (d),
45.9 (d), 38.5 (t), 37.2 (t), 33.0 (t), 24.5 (q), 20.3
(q), 12.3 (q), 12.2 (q), 9.2 (q)

Example 7 Lancacidin C 8-L-lysine ester ( 16 ) L-Lys hydrochloride (5.0 g) was treated in the same manner as in Example to give Nα, Nε-di-Fmoc-L-Lys ( 14.7
g) was obtained as a white powder. This is DCM (490 ml)
, WSC (5.73 g), HOBt (4.04)
g) was added and the mixture was stirred at room temperature for 3 hours. Next, lancasidin A (12.5 g) and TEA (4.14 ml) were added, and the mixture was stirred at room temperature for 62 hours. Hexane (500 ml),
After adding ethyl acetate (500 ml), the mixture was washed with 10% ammonium chloride aqueous solution (300 ml), 2% aqueous sodium hydrogencarbonate (250 ml × 2) and water (250 ml). The organic layer was dried over anhydrous sodium sulfate, concentrated, and then subjected to silica gel chromatography (1.0 liter), developed with an eluent in which acetone in toluene was sequentially added, and eluted with 10% to 20% acetone fractions. Concentrated, triturated from ether-hexane, and Lancacidin A 8-N
α, Nε-di-Fmoc-L-Lys form ( 15 , 13.7)
g) was obtained as a white powder. Anal. Calcd. For C ₆₃ H ₆₇ N ₃ O ₁₃ C; 70.44, H; 6.29, N; 3.91 Found C; 70.67, H; 6.69, N; 3.75 UV λmax nm (ε); 208 (75,200), 220 ( 49,400), 226 (41,
900), 252 (30,100), 262 (27,400), 288 (6,400), 298 (6,4
00), 298 (7,500) (MeOH) IR νmax (cm ⁻¹ ); 1730, 1710, 1510, 1240 (KBr) Compound 15 (6.0 g) was deprotected in the same manner as in Example 6 to give Lancacidin C8. -L-Lys 2 hydrochloride ( 16 ,
380 mg) was obtained as a white powder. [Α] _D −148 ° (c 0.54, H ₂ O, 21 ° C.) Anal. Calcd. For C ₃₁ H ₄₅ N ₃ O ₈ · 2HCl · 2H ₂ OC; 53.45, H; 7.38, N; 6.03, Cl; 10.18 Found C; 53.22, H; 7.54, N; 6.04, Cl; 10.00 UV λmax nm (ε); 226 (49,000) (MeOH) IR νmax (cm ^-1 ); 3400, 1740, 1700, 1630, 1510 , 1270
(KBr) ¹³ C NMR δ ppm (75MHz, DMSO-d ₆ ) 210.9 (s), 196.4 (s), 170.2 (s), 168.4 (s), 159.6 (s),
136.9 (s), 136.9 (s), 136.1 (d), 135.4 (d), 131.2 (d), 1
26.1 (d), 125.2 (d), 124.9 (d), 77.2 (d), 75.0 (d), 67.8
(d), 56.0 (s), 51.5 (d), 51.1 (d), 45.8 (d), 38.0 (t),
37.2 (t), 33.0 (t), 29.1 (t), 26.0 (t), 24.4 (q), 21.1
(t), 20.2 (q), 12.3 (q), 12.2 (q), 9.1 (q)

Example 8 Lancacidin C 8- (aziridine-3-carboxylic acid) -2-carboxylate ( 1
8 ) The amino acid ADA (5.20 g) was dissolved in 5% aqueous sodium hydrogen carbonate solution (160 ml), dioxane (80 ml) was added, and the mixture was cooled to 0 ° C. A 9-fluorenylmethyl chloroformate (Fmoc-Cl, 11.4 g) / dioxane (80 ml) solution was added dropwise thereto over 20 minutes, and the mixture was stirred at 0 ° C. for 1 hour and further at room temperature for 16 hours. The reaction solution was washed with ether (200 ml x 3), adjusted to pH 3.0, and extracted with ethyl acetate (300 ml x 3). After drying over anhydrous sodium sulfate, it was concentrated and Fmoc-ADA (7.74 g)
Was obtained as a white powder. This compound (9.14 g) was added to D
Dissolved in CM (300 ml), WSC (5.96 g), H
OBt (4.20 g) was added and the mixture was stirred at room temperature for 3 hours. Next, Lancacidin A (6.51 g) and TEA (4.34 g)
ml) was added and the mixture was stirred at room temperature for 65 hours. After adding hexane (300 ml) and ethyl acetate (300 ml) to the reaction solution,
It was washed with water (300 ml) and saturated saline (300 ml), dried over anhydrous sodium sulfate and concentrated to obtain a crude substance. This was chromatographed on silica gel, developed with an eluent in which methanol in chloroform was sequentially added, and Lancascidin A 8-F was extracted from 2% and 4% methanol elution fractions.
moc-ADA ( 17 , 7.08 g) was obtained as a white powder. Anal. Calcd. For C ₄₆ H ₄₈ N ₂ O ₁₃・ 2.5H ₂ OC; 62.65, H; 6.06, N; 3.18 Found C; 62.53, H; 5.82, N; 3.36 UV λmax nm (ε); 204 (89,100 ), 218 (62,600), 226 (54,
200), 250 (31,300), 262 (28,000), 286 (10,800), 298 (9,
500) (MeOH) IR νmax (cm ^-1 ); 1730, 1710, 1690, 1620, 1250 (KBr) Compound 17 (1.0 g) was dissolved in DCM (90 ml),
It was cooled to 0 ° C., PIP (10 ml) was added, and the mixture was stirred at 0 ° C. for 40 minutes. Add ethyl acetate (400 ml) to this,
After washing with 0.5N aqueous hydrochloric acid (240 ml) and 0.1% sodium hydrogen carbonate (100 ml × 2), extraction with 0.1% aqueous sodium hydrogen carbonate (100 ml) and 0.2% aqueous sodium hydrogen carbonate (100 ml) did. This was adjusted to pH 5.0 and then concentrated, and the organic solvent was distilled off (300 ml). Esterase I (200 mg) was added to this, and the mixture was stirred at 4 ° C for 16 hours and then adjusted to pH 5.0.
I (200 mg) was added, and the mixture was stirred at room temperature for 3 hours. After adding acetone (1.44 liters) and centrifuging, concentrating the supernatant and distilling acetone off, adjusting the pH to 3.0, extracting with ethyl acetate (150 ml x 2), washing with water, and drying with anhydrous sodium sulfate. After that, it was concentrated and Lancacidin C 8-ADA.
( 18 , 315 mg) was obtained as a white powder. A part of this (300 mg) was dissolved in ethyl acetate (100 ml),
After extraction with 044% sodium hydrogen carbonate (100 ml), p
After adjusting to H6.0 and concentrating and distilling off ethyl acetate, it was chromatographed on Amberlite XAD-II (30 ml), washed with water (120 ml), and 20% acetonitrile water (1 ml).
20 ml), and the eluate was concentrated and lyophilized to give compound 1
The Na salt of 8 (100 mg) was obtained as a white powder. [Α] _D -84.2 ° (c 0.53, H ₂ O, 21
℃) Anal. Calcd. For C ₂₉ H ₃₃ N ₂ O ₁₀ Na ・ 3.5H ₂ OC; 53.13, H; 6.15, N; 4.27, Na; 3.51 Found C; 52.91, H; 6.39, N; 4.42, Na; 3.90 UV λmax nm (ε); 226 (54,500) (MeOH) IR νmax (cm ^-1 ); 3400, 1730, 1710, 1690, 1600, 138
0, 1260 (KBr) ¹³ C NMR δ ppm (75MHz, DMSO-d ₆ ) 210.9 (s), 196.4 (s), 171.0 (s), 170.2 (s), 170.0 (s),
159.6 (s), 137.1 (s), 136.6 (s), 135.6 (d), 135.5 (d), 1
31.0 (d), 125.9 (d), 125.6 (d), 125.4 (d), 75.9 (d), 75.
0 (d), 67.8 (d), 56.1 (s), 51.1 (d), 45.8 (d), 37.2 (t),
33.8 (d), 33.2 (t), 29.5 (d), 24.4 (q), 20.2 (q), 12.2
(q), 12.2 (q), 9.1 (q)

Example 9 Lancacidin C 8-D-alanine ester ( 20 ) Fmoc-D-Ala prepared in the same manner as in Example 6.
Dissolve (5.0 g) in DCM (125 ml) and use WSC
(3.39 g) and HOBt (2.39 g) were added, and the mixture was stirred at room temperature for 3 hours. Next, Lancacidin A (4.03)
g) and TEA (2.45 ml) were added, and the mixture was stirred at room temperature for 4 hours. Hexane (150 ml) and ethyl acetate (1
After adding 50 ml), 10% ammonium chloride aqueous solution (150 ml) and 2% aqueous sodium hydrogen carbonate solution (150 ml ×)
2) and washed with water (100 ml), respectively. 2 organic layers
After standing for a day, the precipitated crystals were filtered to give lancasidin A8-Fmoc-D-Ala ( 19 , 5.3 as colorless crystals.
2 g) was obtained. Anal. Calcd. For C ₄₅ H ₅₀ N ₂ O ₁₁・ 0.25H ₂ OC; 67.61, H; 6.37, N; 3.50 Found C; 67.51, H; 6.34, N; 3.60 UV λmax nm (ε); 208 (65,000 ), 220 (57,600), 226 (54,
500), 250 (31,600), 262 (15,900), 288 (4,800), 298 (5,6
00) (MeOH) IR νmax (cm ^-1 ); 1730, 1710, 1690, 1500, 1250 (KBr) Compound 19 (3.0 g) was dissolved in DCM (135 ml), cooled to 0 ° C., and PIP (15 ml). ) Is added and 4 at 0 ° C.
Stir for hours. Add ethyl acetate (300 ml) to it,
After washing with 0.50N hydrochloric acid water (300ml), 0.019N
After extraction with hydrochloric acid water (200 ml × 2), the pH was adjusted to 6.0, and the mixture was washed with hexane (100 ml × 4). pH 5.0
After adjusting to, concentrate and evaporate the organic solvent, then esterase I
(160 mg) was added, and the mixture was stirred at 4 ° C for 16 hours and then pH was adjusted again
It was adjusted to 5.0 and stirred at room temperature for 2 hours. Acetone (1.0 liter) was added thereto, the mixture was centrifuged, the supernatant was concentrated, 4% aqueous sodium hydrogen carbonate solution (200 ml) was added, the pH was adjusted to 8.6 with aqueous sodium hydroxide solution, and then ethyl acetate ( The mixture was extracted with 150 ml × 3), and the ethyl acetate layer was washed with saturated saline (150 ml). Next, it was re-extracted with 0.023N hydrochloric acid water (250 ml), adjusted to pH 4.0, concentrated and distilled off ethyl acetate, and then XAD-II (150 ml).
Subjected to chromatography, washed with water (600 ml), obtained was eluted with 20% aqueous acetonitrile (600 ml), concentrated, and lyophilized, lankacidin C 8-D-Ala hydrochloride (20, 840 mg) as a white powder Was given. [Α] _D −186 ° (c 0.55, H ₂ O, 21 ° C.) Anal. Calcd. For C ₂₈ H ₃₈ N ₂ O ₈ · HCl · 1.7H ₂ OC; 56.27, H; 7.15, N; 4.69 , Cl; 5.93 Found C; 56.10, H; 7.15, N; 4.65, Cl; 6.24 UV λmax nm (ε); 226 (44,700) (MeOH) IR νmax (cm ^-1 ); 3400, 1740, 1700, 1680, 1630, 151
0, 1260 (KBr) ¹³ C NMR δ ppm (75MHz, DMSO-d ₆ ) 210.9 (s), 196.4 (s), 170.2 (s), 169.1 (s), 159.6 (s),
136.9 (s), 136.9 (s), 135.9 (d), 135.4 (d), 131.2 (d), 1
26.0 (d), 125.2 (d), 124.9 (d), 77.2 (d), 75.0 (d), 67.8
(d), 56.0 (s), 51.1 (d), 47.8 (d), 45.8 (d), 37.1 (t),
33.0 (t), 24.4 (q), 20.2 (q), 15.6 (q), 12.2 (q), 12.2
(q), 9.1 (q)

Example 10 Lancacidin C 8-L-alanine ester ( 22 ) In the same manner as in Example 9, Fmoc-L-Ala (5.0 g) was dissolved in DCM (125 ml) to give WSC (3.39 g),
HOBt (2.39 g) was added, and further lancasidin A was added.
(4.03 g) and TEA (2.45 ml) were added and condensed,
After post-treatment, the organic layer was concentrated, chromatographed on silica gel and developed with an eluent of sequential addition of acetone in toluene. From the 7% and 10% acetone eluate fractions, Lancascidin A 8-Fmoc-L-Ala. ( 21 , 5.8
5 g) was obtained as a white powder. Anal. Calcd. For C ₄₅ H ₅₀ N ₂ O ₁₁・ 0.5C ₇ H ₈ C; 69.27, H; 6.47, N; 3.33 Found C; 69.31, H; 6.59, N; 3.36 UV λmax nm (ε); 208 (67,400), 220 (56,400), 226 (53,
400), 250 (30,200), 262 (21,900), 288 (4,900), 298 (5,600) (MeOH) IR νmax (cm ^-1 ); 1730, 1710, 1690, 1510, 1240 (KBr) Compound 21 (4. 0 g) was dissolved in DCM (180 ml), cooled to 0 ° C., PIP (20 ml) was added, and the mixture was added at 0 ° C. to 4
Stir for hours. After the post-treatment, esterase I reaction was performed in the same manner as in Example 9 to give lancascidin C 8-L-Ala.
The hydrochloride salt ( 22 , 1.26 g) was obtained as a white powder. [Α] _D −178 ° (c 0.53, H ₂ O, 21 ° C.) Anal. Calcd. For C ₂₈ H ₃₈ N ₂ O ₈ HCl.2H ₂ OC; 55.76, H; 7.19, N; 4.64, Cl; 5.88 Found C; 55.71, H; 7.10, N; 4.60, Cl; 6.38 UV λmax nm (ε); 226 (45,800) (MeOH) IR λmax (cm ^-1 ); 3400, 1740, 1700, 1680, 1630 , 151
0, 1260 (KBr) ¹³ C NMR δ ppm (75MHz, DMSO-d ₆ ) 210.9 (s), 196.4 (s), 170.2 (s), 169.1 (s), 159.6 (s),
136.9 (s), 136.9 (s), 136.1 (d), 135.4 (d), 131.2 (d), 1
26.1 (d), 125.2 (d), 124.9 (d), 77.2 (d), 75.0 (d), 67.8
(d), 56.0 (s), 51.1 (d), 47.8 (d), 45.8 (d), 37.2 (t),
33.0 (t), 24.4 (q), 20.2 (q), 15.5 (q), 12.2 (q), 12.2
(q), 9.1 (q)

Example 11 Lancacidin C 8-L-leucine ester ( 24 ) In the same manner as in Example 9, Fmoc-L-Leu (5.0 g) was dissolved in DCM (125 ml) to give WSC (2.99 g),
HOBt (2.11g) was added, and further Lancacidin A
(3.55 g) and TEA (2.16 ml) were added and condensed,
After the workup, the organic layer was concentrated, chromatographed on silica gel, developed with eluate successively added toluene acetone, 10% acetone eluted fraction lankacidin A8-Fmoc-L-Leu ( 23, 6 0.0 g) was obtained as a white powder. Anal. Calcd. For C ₄₈ H ₅₆ N ₂ O ₁₁ C; 68.88, H; 6.74, N; 3.35 Found C; 68.96, H; 6.82, N; 3.25 UV λmax nm (ε); 208 (69,100), 220 ( 57,400), 226 (54,4
00), 250 (30,500), 262 (21,900), 288 (5,000), 298 (5,70
0) (MeOH) IR νmax (cm ⁻¹ ); 1730, 1710, 1690, 1510, 1240 (KBr) Compound 23 (4.0 g) was deprotected in the same manner as in Example 9,
Lyophilized to give Lancacidin C 8-L-Leu hydrochloride ( 2
4, 590 mg) was obtained as a white powder. [Α] _D −158 ° (c 0.54, H ₂ O, 21 ° C.) Anal. Calcd. For C ₃₁ H ₄₄ N ₂ O ₈ · HCl · 1.5H ₂ OC; 58.53, H; 7.60, N; 4.40 , Cl; 5.57 Found C; 58.26, H; 7.43, N; 4.23, Cl; 5.54 UV λmax nm (ε); 226 (52,000) (MeOH) IR νmax (cm ^-1 ); 3400, 1740, 1710, 1690, 1510, 1260
(KBr) ¹³ C NMR δ ppm (75MHz, DMSO-d ₆ ) 210.9 (s), 196.4 (s), 170.2 (s), 168.9 (s), 159.6 (s),
136.9 (s), 136.8 (s), 135.9 (d), 135.5 (d), 131.2 (d), 1
26.1 (d), 125.2 (d), 124.9 (d), 77.2 (d), 75.0 (d), 67.8
(d), 56.0 (s), 51.2 (d), 50.5 (d), 45.8 (d), 39.0 (t),
37.2 (t), 33.0 (t), 24.4 (q), 23.7 (q), 22.1 (q), 21.9
(q), 20.2 (q), 12.2 (q), 12.2 (q), 9.1 (q)

Example 12 Lancacidin C 8-D-Leucine Ester ( 26 ) In the same manner as in Example 9, Fmoc-D-Leu (7.80 g).
Was prepared, and a portion of this (5.0 g) was added to DCM (125 ml).
Dissolved in WSC (2.99 g), HOBt (2.11)
g) was added, and lancascidin A (3.55 g) and T were further added.
EA (2.16 ml) was added and condensed, and after the treatment, crystals were crystallized, which were collected by filtration, and Lancacidin A8-Fmoc-D.
-Leu ( 25 , 3.70 g) was obtained as colorless crystals. Anal. Calcd. For C ₄₈ H ₅₆ N ₂ O ₁₁ C; 68.88, H; 6.74, N; 3.35 Found C; 68.76, H; 6.60, N; 3.36 UV λmax nm (ε); 208 (61,000), 220 ( 54,300), 227 (51,
800), 251 (29,300), 264 (20,900), 287 (5,000), 298 (5,9
00) (MeOH) IR νmax (cm ^-1 ); 1730, 1710, 1690, 1510, 1240 (KBr) This compound 25 (3.60 g) was deprotected in the same manner as in Example 9 and then lyophilized to give Lancacidin C. 8-D-Leu hydrochloride ( 26 , 1.15 g) was obtained as a white powder. [Α] _D −168 ° (c 0.54, H ₂ O, 21 ° C.) Anal. Calcd. For C ₃₁ H ₄₄ N ₂ O ₈ · HCl · 1.5H ₂ OC; 58.53, H; 7.60, N; 4.40 , Cl; 5.57 Found C; 58.37, H; 7.75, N; 4.13, Cl; 5.59 UV λmax nm (ε); 226 (51,700) (MeOH) IR νmax (cm ^-1 ); 3400, 1740, 1710, 1690, 1510, 1260
(KBr) ¹³ C NMR δ ppm (75MHz, DMSO-d ₆ ) 210.9 (s), 196.4 (s), 170.2 (s), 168.9 (s), 159.6 (s),
136.9 (s), 136.9 (s), 135.9 (d), 135.4 (d), 131.3 (d), 1
26.0 (d), 125.2 (d), 124.8 (d), 77.2 (d), 75.0 (d), 67.8
(d), 56.0 (s), 51.1 (d), 50.5 (d), 45.8 (d), 39.1 (t),
37.2 (t), 33.0 (t), 24.4 (q), 23.7 (q), 22.1 (q), 21.9
(q), 20.2 (q), 12.2 (q), 12.1 (q), 9.1 (q)

Example 13 Lancacidin C 8-L-proline ester ( 28 ) In the same manner as in Example 9, Fmoc-L-Pro (5.0 g) was dissolved in DCM (125 ml) to give WSC (3.13 g),
HOBt (2.21 g) was added, and the mixture was stirred at room temperature for 3 hours. Next, Lancacidin A (3.72 g), TEA
After adding (2.26 ml) for condensation and post-treatment, the organic layer was concentrated and subjected to silica gel chromatography, developed with an eluent in which acetone in toluene was sequentially added, and 10%
Lancasidin A 8-Fmoc-L from the acetone-eluted fraction
-Pro ( 27 , 3.50 g) was obtained as a white powder. Anal. Calcd. For C ₄₇ H ₅₂ N ₂ O ₁₁ C; 68.76, H; 6.38, N; 3.41 Found C; 68.47, H; 6.61, N; 3.13 UV λmax nm (ε); 208 (72,400), 220 ( 57,700), 226 (63,
400), 251 (35,000), 264 (14,600), 288 (5,700), 298 (6,5
00) (MeOH) IR νmax (cm ⁻¹ ); 1730, 1710, 1690, 1510, 1240 (KBr) Compound 29 (3.0 g) was deprotected in the same manner as in Example 9,
Lyophilize to give Lancacidin C 8-L-Pro Hydrochloride ( 2
8, 647 mg) was obtained as a white powder. [Α] _D −189 ° (c 0.58, H ₂ O, 21 ° C.) Anal. Calcd. For C ₃₀ H ₄₁ N ₂ O ₈ · HCl · 1H ₂ OC; 58.96, H; 7.09, N; 4.58 Found C; 58.91, H; 7.34, N; 4.30 UV λmax nm (ε); 226 (49,500) (MeOH) IR νmax (cm ^-1 ); 3400, 1740, 1700, 1680, 1510, 1260
(KBr) ¹³ C NMR δ ppm (75MHz, DMSO-d ₆ ) 210.9 (s), 196.4 (s), 170.2 (s), 167.9 (s), 159.6 (s),
136.9 (s), 136.9 (s), 136.1 (d), 135.5 (d), 131.2 (d), 1
26.1 (d), 125.2 (d), 124.9 (d), 77.6 (d), 75.0 (d), 67.8
(d), 58.4 (d), 56.0 (s), 51.1 (d), 45.8 (d), 45.0 (t),
37.2 (t), 32.9 (t), 27.7 (t), 24.4 (q), 22.9 (q), 20.2
(q), 12.2 (q), 12.2 (q), 9.1 (q)

[0036]

The compound [I] of the present invention exhibits a strong antibacterial action and is useful as an antibacterial agent against MRSA and the like.

Claims (3)

[Claims] 1. A general formula: [Wherein R ₁ and R ₂ are a hydroxyl group, a lower alkanoyloxy group or an acyloxy group derived from an amino acid, R ₃ is a hydroxyl group, a lower alkanoyloxy group or an acyloxy group derived from an amino acid and R ₃ ′ is hydrogen] , R
₃ and R ₃ 'represent an oxo group, and at least one of R ₁ , R ₂ and R ₃ is an acyloxy group derived from an optically active amino acid] or a salt thereof. 2. The compound according to claim 1, wherein R ₁ is an acyloxy group derived from an optically active amino acid. 3. An antibacterial agent containing the compound according to claim 1.