CN102766198A - Glycopeptide antifungal compound, and preparation method and application thereof - Google Patents

Abstract

Translated fromChinese

本发明提供一种糖肽类抗真菌化合物及其药学上可接受的盐、其制备方法和用途，以及含有该活性成分的药物组合物，所述糖肽类抗真菌化合物的结构通式如下：其中，n等于1，2或3；R为单糖或二糖，所述的单糖为葡萄糖、半乳糖、木糖、鼠李糖、甘露糖、核糖、氨基葡萄糖或乙酰氨基葡萄糖；所述的二糖为麦芽糖或乳糖。本发明化合物与现有临床应用的抗真菌药物相比，具有高效、低毒、抗菌谱广等优点，因此可用于制备抗真菌药物；本发明的制备方法成功地在Caspofungin的伯氨基上引入了不同的糖基，且工艺简单、产率高。

The present invention provides a glycopeptide antifungal compound and its pharmaceutically acceptable salt, its preparation method and application, and a pharmaceutical composition containing the active ingredient. The general structural formula of the glycopeptide antifungal compound is as follows: Wherein, n is equal to 1, 2 or 3; R is monosaccharide or disaccharide, and described monosaccharide is glucose, galactose, xylose, rhamnose, mannose, ribose, glucosamine or acetylglucosamine; The disaccharides are maltose or lactose. Compared with the antifungal drugs of existing clinical application, the compound of the present invention has the advantages of high efficiency, low toxicity, wide antibacterial spectrum, etc., so it can be used to prepare antifungal drugs; the preparation method of the present invention successfully introduces Caspofungin on the primary amino group Different sugar groups, simple process and high yield.

Description

A kind of glycopeptide class antifungal compound and preparation method thereof and application

Technical field

The present invention relates to the medical compounds technical field, specifically, relate to a kind of glycopeptide class antifungal compound and preparation method thereof and application.

Background technology

In recent years; Because the increase of cancer radiation, chemotherapy and organ transplantation and PCI patient number; Being widely used of Broad spectrum antibiotics, cortin and immunosuppressor clinically, and AIDS is popular, and clinical deep fungal infection case is sharply increased; Nearly 300,000 examples of systemic fungal infection patient that the whole world is diagnosed out every year; In the AIDS patient, it is systemic fungal infection (Markus, R. Mucosal and systemic fungal infections in patients with AIDS:Prophylaxis and treatment. that 60% the direct cause of death is arrangedDrugs, 2004,64,1163-1180.).The life and health that the mycosis serious threat is human.In the medicine of treatment deep fungal infection, the nitrogen azole compounds is used the most extensive, and modal clinically at present fluconazole and itraconazole are exactly azole antifungals.But the day by day serious resistance problem of fluconazole has seriously limited its application.Other antifungal drug; Like polyene antibiotics, propylamine, morpholine class and fluorine miazines etc.; There are toxicity, side effect big (like amphotericin B), narrow antimicrobial spectrum more, are prone to recurrence many deficiencies (Owens, J. Antifungal drugs:A helping hand. such as (like 5-flurocytosines)Nat. Rev.Drug. Discov.2005,1,884-885.).Development is efficient, wide spectrum, the anti-deep fungal infection medicine of low toxicity have become a urgent world subject, causes attention both domestic and external day by day.

The seventies in last century, scientist extracts one type of cyclic peptide compound---echinocandin (Echinocandins) (Denning, W. D. Echinocandin antifungal drugs. that can suppress multiple fungal growth breeding from aspergillus fumigatusLancet, 2003,362,1142-1151.).Discover that further the mechanism of action of this compounds suppresses in the fungal cell membrane for passing throughβ-1,3-D-glucan synthase (Wiederhold, N. P.; Lewis, R. E. The echinocandin antifungals:an overview of the pharmacology, spectrum and clinical efficacy.Expert. Opin. Investig. Drugs. 2003,12,1313-1333.), make the staple of fungal cell wallβ-1; The biosynthesis block of 3-D-VISOSE, and finally cause the cytoskeleton of fungi to form and dead (see figure 1), such antifungal compound and amphotericin B and azole antifungal medicine medicine do not have cross resistance; Because the mammal cell does not contain cell walls, does not produceβ-1; The 3-D-glucan synthase is so this type of drug selectivity is high, effect is powerful, toxicity is lower, adds easy to use, better tolerance; Have broad application prospects, at present the research of this compounds has been become the focus and new direction of current searching novel anti fungi-medicine.

At present; Three echinocandin-class antifungal drug listings have been arranged; Be respectively Caspofungin (Caspofungin, trade(brand)name: MK 0991, Cancidas), Mi Kafen is clean and anidulafungin; Such medicine is mainly used in invasive aspergillosis and moniliosis (Yalaz, the M. of treatment refractory clinically; Akisu, M.; Hilmioglu, S.; Calkavur, S.; Cakmak, B.; Kultursay, N. Successful caspofungin treatment of multidrug resistant Candida parapsilosis septicaemia in an extremely low birth weight neonate.Mycoses. 2006,49,242-245.).Wherein, Caspofungin is released in calendar year 2001 by Merck & Co., Inc., is the echinocandin class medicine of first listing, and its chemical structural formula is shown in the following figure:

Because Most amino-acids is alpha-non-natural amino acid in the structure of Caspofungin, so its complete synthesis difficulty is very big, does not see the complete synthesis report of this compound so far as yet.Merck & Co., Inc. reported by echinocandin B in 2007₀Be raw material, realized semi-synthetic (Leonard, the W. R. of Caspofungin through three-step reaction; Belyk, K. M.; Conlon, D. A.; Bender, D. R.; DiMichele, L. M.; Liu, J.; Hughes, D. L. Synthesis of the antifungal beta-1,3-glucan synthase inhibitor CANCIDAS (caspofungin acetate) from pneumocandin B₀.J. Org. Chem.2007,72,2335-2343.).In addition, to the segmental synthetic part report that also has of the part in the Caspofungin, for example, Leonard took the lead in realizing the chirality of the longer chain fatty acid in the Caspofungin structure synthetic (Leonard, W. R. in 2002; Belyk, K. M.; Bender, D. R.; Conlon, D. A.; Hughes, D. L.; Reider P. J. Determination of the Relative and Absolute Configuration of the Dimethylmyristoyl Side Chain of Pneumocandin B0 by Asymmetric Synthesis.Org. Lett.2002,4,4201-4204.).

Although the Caspofungin as the echinocandin class antifungal drug has good anti-mycotic activity and has been applied to clinical anti-infective therapy.But still there is following defective in it: at first, Caspofungin is to neural genotoxic potential, mainly is because it good fat-solublely causes it to see through due to the hemato encephalic barrier easily; Secondly, the Caspofungin poorly water-soluble, need process acetate usually can patent medicine; Once more, the acetate of this medicine is deliquescence very easily, and poor stability has limited the variation of its formulation, and the formulation of this medicine has only lyophilized powder a kind of at present.More than these issues limit its clinical application (Pacetti, S.A.; Gelone, S. P. Caspofungin acetate for treatment of invasive fungal infections.Ann. Pharmacother.2003,37,90-98.).Therefore, the structure of Caspofungin being optimized, reducing that it is fat-soluble, it is water-soluble to improve, to improve its stability and to reduce spinoff, promote its applying clinically, is very necessary.But also do not meet similar report at present.

Summary of the invention

The objective of the invention is to deficiency of the prior art, the glycopeptide class antifungal compound and the pharmacy acceptable salt thereof of a kind of efficient, low toxicity, wide spectrum is provided.

One purpose more of the present invention is that a kind of pharmaceutical composition is provided.

Another purpose of the present invention is that the preparation method of a kind of glycopeptide class antifungal compound and pharmacy acceptable salt thereof is provided.

The 4th purpose of the present invention is that the purposes of a kind of glycopeptide class antifungal compound and pharmacy acceptable salt thereof is provided.

For realizing above-mentioned purpose, the technical scheme that the present invention takes is:

A kind of glycopeptide class antifungal compound and pharmacy acceptable salt thereof, described Caspofungin glycosyl derivatives general structure is following:

Wherein, n equals 1,2 or 3; R is monose or disaccharides; Described monose is glucose, semi-lactosi, wood sugar, rhamnosyl, seminose, ribose, GS or acetylglucosamine; Described disaccharides is SANMALT-S or lactose.

Described n equals 3.

Described R is glucose or SANMALT-S.

Described pharmacy acceptable salt class is hydrochloride, vitriol, hydrosulfate, hydrobromate, acetate, oxalate, Citrate trianion or mesylate.

For realizing above-mentioned second purpose, the technical scheme that the present invention takes is:

A kind of pharmaceutical composition, it contains as above arbitrary described glycopeptide class antifungal compound and pharmacy acceptable salt thereof, and contains conventional pharmaceutical carrier.

For realizing above-mentioned the 3rd purpose, the technical scheme that the present invention takes is:

The preparation method of a kind of as above arbitrary described glycopeptide class antifungal compound and pharmacy acceptable salt thereof, described method comprises the following steps:

A) preparation key intermediate glycosyl Acibenzolar: with glycosyls such as glucose, semi-lactosi, wood sugar, rhamnosyl, SANMALT-S and lactose is starting raw material, obtains the glycosyl of full acetyl protection with sodium-acetate back flow reaction in acetic anhydride; Under the condition that trimethyl azide silane and BFEE exist substitution reaction takes place, the acetoxyl group of 1-position is become azido-then; Azido-generates amino back through 10% Pd-C catalytic hydrogenation and under the condition that 4-(N, N-dimethylamino) pyridine exists, is condensed into acid amides with corresponding acid anhydrides (malonic anhydride, Succinic anhydried, Pyroglutaric acid); The carboxyl that exposes out with N, generate corresponding glycosyl Acibenzolar with N-maloyl imines generation ester condensation under the condition that the N-NSC 57182 exists;

B) preparation target compound Caspofungin glycosyl derivatives: the Caspofungin glycosyl compound that condensation generates full acetyl protection takes place in key intermediate glycosyl Acibenzolar and Caspofungin acetate under the triethylamine alkaline condition, and then is that deacetylate protection promptly gets target compound Caspofungin glycosyl derivatives in sodium methylate/methanol solution of 8 at pH.

For realizing above-mentioned the 4th purpose, the technical scheme that the present invention takes is:

The application in preparation treatment fungi infestation medicine of as above arbitrary described glycopeptide class antifungal compound and pharmacy acceptable salt thereof.

The invention has the advantages that:

1, compound glycopeptide class antifungal compound of the present invention and pharmacy acceptable salt thereof are to have introduced different glycosyls on the primary amino in the Caspofungin structure, have solved shortcomings such as Caspofungin poor stability and neurotoxicity are strong; Simultaneously, can strengthen the interaction between it and the target enzyme introducing glycosyl on this Caspofungin molecule, thereby improve anti-mycotic activity.Pharmacological evaluation proves, it is active that compound of the present invention shows stronger inhibition to the part fungi, compares with the antifungal drug of existing clinical application, has advantages such as efficient, low toxicity, has a broad antifungal spectrum, therefore can be used for preparing antifungal drug.

2, preparation method of the present invention has successfully introduced different glycosyls on the primary amino of Caspofungin, prepare the Caspofungin glycosyl derivatives of efficient, low toxicity, has a broad antifungal spectrum, and technology is simple, productive rate is high.

Description of drawings

Accompanying drawing 1 is the formation of fungal cell's skeleton.

Accompanying drawing 2 is compounds1a,1eWith contrast medicine Caspofungin1The stability experiment result: (A) high wet test; (B) high temperature test.

Embodiment

Below in conjunction with accompanying drawing embodiment provided by the invention is elaborated.

The high resolution mass spectrum of embodiment 1 The compounds of this invention 1a-f and nucleus magnetic hydrogen spectrum data

The compounds of this invention1a-fHigh resolution mass spectrum and nucleus magnetic hydrogen spectrum data see table 1 and table 2.

Table 1 The compounds of this invention1a-fThe high resolution mass spectrum table

The compound sequence number	n	R	HR-Q-TOF-MS+
				1a	3		1642.8437
1b	3		1642.8458
				1c	3		1582.8227
1d	3		1610.8553
				1e	3		1966.9497
1f	3		1966.9497

Table 2 The compounds of this invention1a-fThe nucleus magnetic hydrogen spectrum data sheet

The preparation of embodiment 2 The compounds of this invention 1a-f

The compounds of this invention1a-fPreparation can slightly be divided into for two steps:

A) preparation key intermediate glycosyl Acibenzolar7a-f, reaction scheme is following:

B) preparation target compound1a-f, reaction scheme is following:

The concrete preparation method of The compounds of this invention 1a-f is following:

One, preparation midbody acetyl glucosides (3a-f)

(1) preparation 1,2,3,4,6-five-O-ethanoyl-β-D-glucopyranoside (3a)

Willβ-D-Glucopyranose2a(40 g, 222 mmol) and sodium-acetate (30g, 330 mmol) place 500 mL three-necked bottles, add diacetyl oxide (350 mL, 3.71 mol).The reaction mixture mechanical stirring, behind 170 ℃ of back flow reaction 6 h, the room temperature cooling is poured in the mixture of ice and water (1.5 L) mixture and stirred overnight.Next day, observation had brown solid to separate out, and filtered, and filter cake gets midbody with recrystallizing methanol and with decolorizing with activated carbon3a,Be the white powder solid, weigh 63 g, yield 72%.

(2) preparation 1,2,3,4,6-five-O-ethanoyl-β-D-galactopyranoside (3b)

Willβ-D-galactopyranose2b(40 g, 222 mmol) and sodium-acetate (30g, 330 mmol) place 500 mL three-necked bottles, add diacetyl oxide (350 mL, 3.71 mol).The reaction mixture mechanical stirring, behind 170 ℃ of back flow reaction 6 h, the room temperature cooling is poured in the mixture of ice and water (1.5 L) mixture and stirred overnight.Next day, observation had brown solid to separate out, and filtered, and filter cake gets midbody with recrystallizing methanol and with decolorizing with activated carbon3b,Be the white powder solid, weigh 65 g, yield 75%.

(3) preparation 1,2,3,4 – four-O-ethanoyl-β-D-xylopyranoside (3c)

Willβ-D-xylopyranose2c(34 g, 222 mmol) and sodium-acetate (30g, 330 mmol) place 500 mL three-necked bottles, add diacetyl oxide (350 mL, 3.71 mol).The reaction mixture mechanical stirring, behind 170 ℃ of back flow reaction 6 h, the room temperature cooling is poured in the mixture of ice and water (1.5 L) mixture and stirred overnight.Next day, observation had brown solid to separate out, and filtered, and filter cake gets midbody with recrystallizing methanol and with decolorizing with activated carbon3c,Be the white powder solid, weigh 51 g, yield 72 %.

(4) preparation 1,2,3,4-four-O-ethanoyl-β-D-pyrans rhamnoside (3d)

Willβ-D-pyrans rhamnosyl2d(36 g, 222 mmol) and sodium-acetate (30 g, 330 mmol) place 500 mL three-necked bottles, add diacetyl oxide (350 mL, 3.71 mol).The reaction mixture mechanical stirring, behind 170 ℃ of back flow reaction 6 h, the room temperature cooling is poured in the mixture of ice and water (1.5 L) mixture and stirred overnight.Next day, observation had brown solid to separate out, and filtered, and filter cake gets midbody with recrystallizing methanol and with decolorizing with activated carbon3d,Be the white powder solid, weigh 55 g, yield 74 %.

(5) preparation 2,3,4,6-four-O-ethanoyl-β-D-glucopyranosyl-(1-4)-1,2,3,6-four-O-ethanoyl-β-D-glucopyranoside (3e)

Willβ-D-pyrans SANMALT-S2e(76 g, 222 mmol) and sodium-acetate (30 g, 330 mmol) place 500 mL three-necked bottles, add diacetyl oxide (350 mL, 3.71 mol).The reaction mixture mechanical stirring, behind 170 ℃ of back flow reaction 6 h, the room temperature cooling is poured in the mixture of ice and water (1.5 L) mixture and stirred overnight.Next day, observation had brown solid to separate out, and filtered, and filter cake gets midbody with recrystallizing methanol and with decolorizing with activated carbon3e,Be the white powder solid, weigh 107 g, yield 71 %.

(6) preparation 2,3,4,6-four-O-ethanoyl-β-D-galactopyranose base-(1-4)-1,2,3,6-four-O-ethanoyl-β-D-glucopyranoside (3f)

Willβ-D-pyrans lactose2f(76 g, 222 mmol) and sodium-acetate (30g, 330 mmol) place 500 mL three-necked bottles, add (350 mL, 3.71 mol) diacetyl oxide.The reaction mixture mechanical stirring, behind 170 ℃ of back flow reaction 6 h, the room temperature cooling is poured in the mixture of ice and water (1.5 L) mixture and stirred overnight.Next day, observation had brown solid to separate out, and filtered, and filter cake gets midbody with recrystallizing methanol and with decolorizing with activated carbon3f,Be the white powder solid, weigh 102 g, yield 68 %.

Two, preparation midbody 1-azido-acetyl glucosides (4a-f)

(1) preparation 1-azido--2,3,4,6-four-O-ethanoyl-β-D-glucopyranoside (4a)

With acetylizad glucose3a(10 g, 25.62 mmol) and 4A molecular sieve 10 g place 250 mL eggplant-shape bottles, vacuumize, and argon shield adds anhydrous methylene chloride 100 mL under the condition of ice bath, stir 10 min after, dropwise add trimethyl azide silane (TMSN₃, 8.5 mL, 64.05 mmol), after continuing to stir 30 min, dropwise add BFEE (BF₃.Et₂O, 8.0 mL, 64.05 mmol), after continuing to stir 1 h, remove ice bath, after stirring at room is reacted 8 h, TLC (sherwood oil: inspection ETHYLE ACETATE=1: 1), react complete, so suction filtration, filter cake is used washed with dichloromethane, pours filtrating into saturated NaHCO then₃In, be stirred to and do not produce till the bubble, tell organic layer, organic layer is used saturated NaHCO again₃Respectively wash once anhydrous Na with saturated NaCl₂SO₄Drying behind the concentrating under reduced pressure, is crossed purification by silica gel column chromatography, and developping agent is sherwood oil and ETHYLE ACETATE, gets midbody 4a behind the purifying, is the white powder solid, weighs 7.6 g, yield 79%.

(2) preparation 1-azido--2,3,4,6-four-O-ethanoyl-β-D-galactopyranoside (4b)

With acetylizad semi-lactosi3b(10 g, 25.62 mmol) and 4A molecular sieve 10 g place 250 mL eggplant-shape bottles, vacuumize, and argon shield adds anhydrous methylene chloride 100 mL under the condition of ice bath, stir 10 min after, dropwise add trimethyl azide silane (TMSN₃, 8.5 mL, 64.05 mmol), after continuing to stir 30 min, dropwise add BFEE (BF₃Et₂O, 8.0 mL, 64.05 mmol), after continuing to stir 1 h, remove ice bath, after stirring at room is reacted 8 h, TLC (sherwood oil: inspection ETHYLE ACETATE=1: 1), react complete, so suction filtration, filter cake is used washed with dichloromethane, pours filtrating into saturated NaHCO then₃In, be stirred to and do not produce till the bubble, tell organic layer, organic layer is used saturated NaHCO again₃Respectively wash once anhydrous Na with saturated NaCl₂SO₄Drying behind the concentrating under reduced pressure, is crossed purification by silica gel column chromatography, and developping agent is sherwood oil and ETHYLE ACETATE, gets midbody 4b behind the purifying, is the white powder solid, weighs 7.8 g, yield 82 %.

(3) preparation 1-azido--2,3,4-three-O-ethanoyl-β-D-xylopyranoside (4c)

With acetylizad wood sugar3c(8 g, 25.62 mmol) and 4A molecular sieve 10 g place 250 mL eggplant-shape bottles, vacuumize, and argon shield adds anhydrous methylene chloride 100 mL under the condition of ice bath, stir 10 min after, dropwise add trimethyl azide silane (TMSN₃, 8.5 mL, 64.05 mmol), after continuing to stir 30 min, dropwise add BFEE (BF₃.Et₂O, 8.0 mL, 64.05 mmol), after continuing to stir 1 h, remove ice bath, after stirring at room is reacted 8 h, TLC (sherwood oil: inspection ETHYLE ACETATE=1: 1), react complete, so suction filtration, filter cake is used washed with dichloromethane, pours filtrating into saturated NaHCO then₃In, be stirred to and do not produce till the bubble, tell organic layer, organic layer is used saturated NaHCO again₃Respectively wash once anhydrous Na with saturated NaCl₂SO₄Drying behind the concentrating under reduced pressure, is crossed purification by silica gel column chromatography, and developping agent is sherwood oil and ETHYLE ACETATE, gets midbody 4c behind the purifying, is the white powder solid, weighs 5.9 g, yield 77 %.

(4) preparation 1-azido--2,3,4-three-O-ethanoyl-β-D-pyrans rhamnoside (4d)

With acetylizad rhamnosyl3d(8.5 g, 25.62 mmol) and 4A molecular sieve 10 g place 250 mL eggplant-shape bottles, vacuumize, and argon shield adds anhydrous methylene chloride 100 mL under the condition of ice bath, stir 10 min after, dropwise add trimethyl azide silane (TMSN₃, 8.5 mL, 64.05 mmol), after continuing to stir 30 min, dropwise add BFEE (BF₃Et₂O, 8.0 mL, 64.05 mmol), after continuing to stir 1 h, remove ice bath, after stirring at room is reacted 8 h, TLC (sherwood oil: inspection ETHYLE ACETATE=1: 1), react complete, so suction filtration, filter cake is used washed with dichloromethane, pours filtrating into saturated NaHCO then₃In, be stirred to and do not produce till the bubble, tell organic layer, organic layer is used saturated NaHCO again₃Respectively wash once anhydrous Na with saturated NaCl₂SO₄Drying behind the concentrating under reduced pressure, is crossed purification by silica gel column chromatography, and developping agent is sherwood oil and ETHYLE ACETATE, gets midbody 4d behind the purifying, is the white powder solid, weighs 6.3 g, yield 78 %.

(5) preparation 1-azido--2,3,4,6-four-O-ethanoyl-β-D-glucopyranosyl-(1-4)-2,3,6-three-O-ethanoyl-β-D-glucopyranoside (4e)

With acetylizad SANMALT-S3e(17.5 g, 25.62 mmol) and 4A molecular sieve 10 g place 250 mL eggplant-shape bottles, vacuumize, and argon shield adds anhydrous methylene chloride 100 mL under the condition of ice bath, stir 10 min after, dropwise add trimethyl azide silane (TMSN₃, 8.5 mL, 64.05 mmol), after continuing to stir 30 min, dropwise add BFEE (BF₃Et₂O, 8.0 mL, 64.05 mmol), after continuing to stir 1 h, remove ice bath, after stirring at room is reacted 8 h, TLC (sherwood oil: inspection ETHYLE ACETATE=1: 1), react complete, so suction filtration, filter cake is used washed with dichloromethane, pours filtrating into saturated NaHCO then₃In, be stirred to and do not produce till the bubble, tell organic layer, organic layer is used saturated NaHCO again₃Respectively wash once anhydrous Na with saturated NaCl₂SO₄Drying behind the concentrating under reduced pressure, is crossed purification by silica gel column chromatography, and developping agent is sherwood oil and ETHYLE ACETATE, gets midbody 4e behind the purifying, is the white powder solid, weighs 13.7 g, yield 81 %.

(6) preparation 1-azido--2,3,4,6-four-O-ethanoyl-β-D-galactopyranose base-(1-4)-2,3,6-three-O-ethanoyl-β-D-glucopyranoside (4f)

With acetylizad lactose3f(17.5 g, 25.62 mmol) and 4A molecular sieve 10 g place 250 mL eggplant-shape bottles, vacuumize, and argon shield adds anhydrous methylene chloride 100 mL under the condition of ice bath, stir 10 min after, dropwise add trimethyl azide silane (TMSN₃, 8.5 mL, 64.05 mmol), after continuing to stir 30 min, dropwise add BFEE (BF₃Et₂O, 8.0 mL, 64.05 mmol), after continuing to stir 1 h, remove ice bath, after stirring at room is reacted 8 h, TLC (sherwood oil: inspection ETHYLE ACETATE=1: 1), react complete, so suction filtration, filter cake is used washed with dichloromethane, pours filtrating into saturated NaHCO then₃In, be stirred to and do not produce till the bubble, tell organic layer, organic layer is used saturated NaHCO again₃Respectively wash once anhydrous Na with saturated NaCl₂SO₄Drying behind the concentrating under reduced pressure, is crossed purification by silica gel column chromatography, and developping agent is sherwood oil and ETHYLE ACETATE, gets midbody 4f behind the purifying, is the white powder solid, weighs 13.1 g, yield 77 %.

Three, preparation midbody 1-glycyl glucosides (5a-f)

(1) preparation 1-amino-2,3,4,6-four-O-ethanoyl-β-D-glucopyranoside (5a)

The nitrine acetyl glucosamine that drying is good4a(3.0 g, 8.04 mmol) put in the 100 mL eggplant-shape bottles, after adding anhydrous methylene chloride 6 mL make it to dissolve fully, add 10% Pd-C (450 mg again; W/w=15%) and methyl alcohol 30 mL, vacuumize, charge into hydrogen; Behind stirring at room 1 h, and TLC (sherwood oil: inspection ETHYLE ACETATE=1: 1), react complete; So suction filtration after filtrating concentrates, gets the bullion midbody5a, not purified, directly carry out next step reaction.

(2) preparation 1-amino-2,3,4,6-four-O-ethanoyl-β-D-galactopyranoside (5b)

The nitrine acetyl semi-lactosi that drying is good4b(3.0 g, 8.04 mmol) put in the 100 mL eggplant-shape bottles, after adding anhydrous methylene chloride 6 mL make it to dissolve fully, add 10% Pd-C (450 mg again; W/w=15%) and methyl alcohol 30 mL, vacuumize, charge into hydrogen; Behind the stirring at room 1h, and TLC (sherwood oil: inspection ETHYLE ACETATE=1: 1), react complete; So suction filtration after filtrating concentrates, gets the bullion midbody5b, not purified, directly carry out next step reaction.

(3) preparation 1-amino-2,3,4-three-O-ethanoyl-β-D-xylopyranoside (5c)

The nitrine acetyl wood sugar that drying is good4c(3.0 g, 9.96 mmol) put in the 100 mL eggplant-shape bottles, after adding anhydrous methylene chloride 6 mL make it to dissolve fully, add 10% Pd-C (450 mg again; W/w=15%) and methyl alcohol 30 mL, vacuumize, charge into hydrogen; Behind stirring at room 1 h, and TLC (sherwood oil: inspection ETHYLE ACETATE=1: 1), react complete; So suction filtration after filtrating concentrates, gets the bullion midbody5c, not purified, directly carry out next step reaction.

(4) preparation 1-amino-2,3,4 ,-three-O-ethanoyl-β-D-pyrans rhamnoside (5d)

The nitrine acetyl rhamnosyl that drying is good4d(3.0 g, 9.52 mmol) put in the 100 mL eggplant-shape bottles, after adding anhydrous methylene chloride 6 mL make it to dissolve fully, add 10% Pd-C (450 mg again; W/w=15%) and methyl alcohol 30 mL, vacuumize, charge into hydrogen; Behind stirring at room 1 h, and TLC (sherwood oil: inspection ETHYLE ACETATE=1: 1), react complete; So suction filtration after filtrating concentrates, gets the bullion midbody5d, not purified, directly carry out next step reaction.

(5) preparation 1-amino-2,3,4,6-four-O-ethanoyl-β-D-glucopyranosyl-(1-4)-2,3,6-three-O-ethanoyl-β-D-glucopyranoside (5e)

The nitrine acetyl SANMALT-S that drying is good4e(3.0 g, 4.53 mmol) put in the 100 mL eggplant-shape bottles, after adding anhydrous methylene chloride 6 mL make it to dissolve fully, add 10% Pd-C (450 mg again; W/w=15%) and methyl alcohol 30 mL, vacuumize, charge into hydrogen; Behind stirring at room 1 h, and TLC (sherwood oil: inspection ETHYLE ACETATE=1: 1), react complete; So suction filtration after filtrating concentrates, gets the bullion midbody5e, not purified, directly carry out next step reaction.

(6) preparation 1-amino-2,3,4,6-four-O-ethanoyl-β-D-galactopyranose base-(1-4)-2,3,6-three-O-ethanoyl-β-D-glucopyranoside (5f)

The nitrine acetyl lactose that drying is good4f(3.0 g, 4.53 mmol) put in the 100 mL eggplant-shape bottles, after adding anhydrous methylene chloride 6 mL make it to dissolve fully, add 10% Pd-C (450 mg again; W/w=15%) and methyl alcohol 30 mL, vacuumize, charge into hydrogen; Behind stirring at room 1 h, and TLC (sherwood oil: inspection ETHYLE ACETATE=1: 1), react complete; So suction filtration after filtrating concentrates, gets the bullion midbody5f, not purified, directly carry out next step reaction.

Four, preparation midbody glycosyl acid (6a-f)

(1) preparation midbody glycosyl acid (6a)

With the dry good bullion glycyl glucose of step 35aWith the 4-of catalytic amount (N,N-dimethylamino) (DMAP 0.050g) puts in the 100 mL eggplant-shape bottles pyridine, after adding anhydrous methylene chloride 12 mL make it dissolving; Add anhydrous pyridine 3 mL again, behind stirring 10 min, add Pyroglutaric acid under the condition of ice bath; Continue to stir 1 h recession deicing and bathe, after stirring at room is reacted 3 h, TLC (methylene dichloride: inspection methyl alcohol=10: 1); React complete, then in reaction solution, add methylene dichloride 60 mL, with the HCl solution washing of 30 mL 5%; Anhydrous magnesium sulfate drying concentrates the back and uses re-crystallizing in ethyl acetate, gets midbody6a,Be a white powder, heavy 1.68g, two step total recovery 45 %.

(2) preparation midbody glycosyl acid (6b)

With the dry good bullion glycyl glucose of step 35bWith the 4-of catalytic amount (N,N-dimethylamino) (DMAP 0.050g) puts in the 100 mL eggplant-shape bottles pyridine, after adding anhydrous methylene chloride 12 mL make it dissolving; Add anhydrous pyridine 3 mL again, behind stirring 10 min, add Pyroglutaric acid under the condition of ice bath; Continue to stir 1 h recession deicing and bathe, after stirring at room is reacted 3 h, TLC (methylene dichloride: inspection methyl alcohol=10: 1); React complete, then in reaction solution, add methylene dichloride 60 mL, with the HCl solution washing of 30 mL 5%; Anhydrous magnesium sulfate drying concentrates the back and uses re-crystallizing in ethyl acetate, gets midbody6b,Be a white powder, weigh 1.81 g, two step total recovery 49 %.

(3) preparation midbody glycosyl acid (6c)

With the dry good bullion glycyl glucose of step 35cWith the 4-of catalytic amount (N,N-dimethylamino) (DMAP 0.050g) puts in the 100 mL eggplant-shape bottles pyridine, after adding anhydrous methylene chloride 12 mL make it dissolving; Add anhydrous pyridine 3 mL again, behind stirring 10 min, add Pyroglutaric acid under the condition of ice bath; Continue to stir 1 h recession deicing and bathe, after stirring at room is reacted 3 h, TLC (methylene dichloride: inspection methyl alcohol=10: 1); React complete, then in reaction solution, add methylene dichloride 60 mL, with the HCl solution washing of 30 mL 5%; Anhydrous magnesium sulfate drying concentrates the back and uses re-crystallizing in ethyl acetate, gets midbody6c,Be a white powder, weigh 1.82 g, yield 47 %.

(4) preparation midbody glycosyl acid (6d)

With the dry good bullion glycyl glucose of step 35dWith the 4-of catalytic amount (N,N-dimethylamino) (DMAP 0.050g) puts in the 100 mL eggplant-shape bottles pyridine, after adding anhydrous methylene chloride 12 mL make it dissolving; Add anhydrous pyridine 3 mL again, behind stirring 10 min, add Pyroglutaric acid under the condition of ice bath; Continue to stir 1 h recession deicing and bathe, after stirring at room is reacted 3 h, TLC (methylene dichloride: inspection methyl alcohol=10: 1); React complete, then in reaction solution, add methylene dichloride 60 mL, with the HCl solution washing of 30 mL 5%; Anhydrous magnesium sulfate drying concentrates the back and uses re-crystallizing in ethyl acetate, gets midbody6d,Be a white powder, weigh 1.77 g, yield 46 %.

(5) preparation midbody glycosyl acid (6e)

With the dry good bullion glycyl glucose of step 35eWith the 4-of catalytic amount (N,N-dimethylamino) (DMAP 0.050g) puts in the 100 mL eggplant-shape bottles pyridine, after adding anhydrous methylene chloride 12 mL make it dissolving; Add anhydrous pyridine 3 mL again, behind stirring 10 min, add Pyroglutaric acid under the condition of ice bath; Continue to stir 1 h recession deicing and bathe, after stirring at room is reacted 3 h, TLC (methylene dichloride: inspection methyl alcohol=10: 1); React complete, then in reaction solution, add methylene dichloride 60 mL, with the HCl solution washing of 30 mL 5%; Anhydrous magnesium sulfate drying concentrates the back and uses re-crystallizing in ethyl acetate, gets midbody6e,Be a white powder, heavy 1.70g, yield 50 %.

(6) preparation midbody glycosyl acid (6f)

With the dry good bullion glycyl glucose of step 35fWith the 4-of catalytic amount (N,N-dimethylamino) (DMAP 0.050g) puts in the 100 mL eggplant-shape bottles pyridine, after adding anhydrous methylene chloride 12 mL make it dissolving; Add anhydrous pyridine 3 mL again, behind stirring 10 min, add Pyroglutaric acid under the condition of ice bath; Continue to stir 1 h recession deicing and bathe, after stirring at room is reacted 3 h, TLC (methylene dichloride: inspection methyl alcohol=10: 1); React complete, then in reaction solution, add methylene dichloride 60 mL, with the HCl solution washing of 30 mL 5%; Anhydrous magnesium sulfate drying concentrates the back and uses re-crystallizing in ethyl acetate, gets midbody6f,Be a white powder, weigh 1.66 g, yield 49 %.

Five, preparation key intermediate glycosyl Acibenzolar (7a-f)

(1) preparation key intermediate glycosyl Acibenzolar (7a)

With the acid of midbody glycosyl6a(500 mg, 1.084 mmol) and N-maloyl imines (HOSU, 150 mg, 1.303 mmol) and N; N-NSC 57182 (DCC, 206 mg, 1.301 mmol) places 25 mL eggplant-shape bottles, adds methylene dichloride 10 mL and makes it to dissolve fully; Stirring at room, the adularescent solid is separated out behind 10 min, (the methylene dichloride: inspection methyl alcohol=10: 1) of TLC behind 30 min; React complete,, get bullion after filtrating concentrates so filter7a, not purified, directly carry out next step reaction.

(2) preparation key intermediate glycosyl Acibenzolar (7b)

With the acid of midbody glycosyl6b(500 mg, 1.084 mmol) and N-maloyl imines (HOSU, 150 mg, 1.303 mmol) and N; N-NSC 57182 (DCC, 206 mg, 1.301 mmol) places 25 mL eggplant-shape bottles, adds methylene dichloride 10 mL and makes it to dissolve fully; Stirring at room, the adularescent solid is separated out behind 10 min, (the methylene dichloride: inspection methyl alcohol=10: 1) of TLC behind 30 min; React complete,, get bullion after filtrating concentrates so filter7b, not purified, directly carry out next step reaction.

(3) preparation key intermediate glycosyl Acibenzolar (7c)

With the acid of midbody glycosyl6c(422 mg, 1.084 mmol) and N-maloyl imines (HOSU, 150 mg, 1.303 mmol) and N; N-NSC 57182 (DCC, 206 mg, 1.301 mmol) places 25 mL eggplant-shape bottles, adds methylene dichloride 10 mL and makes it to dissolve fully; Stirring at room, the adularescent solid is separated out behind 10 min, (the methylene dichloride: inspection methyl alcohol=10: 1) of TLC behind 30 min; React complete,, get bullion after filtrating concentrates so filter7c, not purified, directly carry out next step reaction.

(4) preparation key intermediate glycosyl Acibenzolar (7d)

With the acid of midbody glycosyl6a(437 mg, 1.084 mmol) and N-maloyl imines (HOSU, 150 mg, 1.303 mmol) and N; N-NSC 57182 (DCC, 206 mg, 1.301 mmol) places 25 mL eggplant-shape bottles, adds methylene dichloride 10 mL and makes it to dissolve fully; Stirring at room, the adularescent solid is separated out behind 10 min, (the methylene dichloride: inspection methyl alcohol=10: 1) of TLC behind 30 min; React complete,, get bullion after filtrating concentrates so filter7d, not purified, directly carry out next step reaction.

(5) preparation key intermediate glycosyl Acibenzolar (7e)

With the acid of midbody glycosyl6e(812 mg, 1.084 mmol) and N-maloyl imines (HOSU, 150 mg, 1.303 mmol) and N; N-NSC 57182 (DCC, 206 mg, 1.301 mmol) places 25 mL eggplant-shape bottles, adds methylene dichloride 10 mL and makes it to dissolve fully; Stirring at room, the adularescent solid is separated out behind 10 min, (the methylene dichloride: inspection methyl alcohol=10: 1) of TLC behind 30 min; React complete,, get bullion after filtrating concentrates so filter7e, not purified, directly carry out next step reaction.

(6) preparation key intermediate glycosyl Acibenzolar (7f)

With the acid of midbody glycosyl6f(812 mg, 1.084 mmol) and N-maloyl imines (HOSU, 150 mg, 1.303 mmol) and N; N-NSC 57182 (DCC, 206 mg, 1.301 mmol) places 25 mL eggplant-shape bottles, adds methylene dichloride 10 mL and makes it to dissolve fully; Stirring at room, the adularescent solid is separated out behind 10 min, (the methylene dichloride: inspection methyl alcohol=10: 1) of TLC behind 30 min; React complete,, get bullion after filtrating concentrates so filter7f, not purified, directly carry out next step reaction.

Six, preparation midbody (9a-f)

(1) preparation midbody (9a)

With commercially available Caspofungin acetate8(50 mg; 0.0412 mmol) place 25 mL eggplant-shape bottles, add each 2 mL of entry and dioxane and make it to dissolve fully, drip triethylamine (0.03 mL under the stirring at room; 0.206 mmol), drip the key intermediate glycosyl Acibenzolar that is dissolved in dioxane behind 10 min again7a(by 90% purity meter, 60 mg, 0.0989 mmol), a complete back room temperature continues to stir 6 h, and the ESI-MS detection reaction is complete, so with getting bullion behind the reaction solution cryoconcentration9a, directly carry out next step reaction.

(2) preparation midbody (9b)

With commercially available Caspofungin acetate8(50 mg; 0.0412 mmol) place 25 mL eggplant-shape bottles, add each 2 mL of entry and dioxane and make it to dissolve fully, drip triethylamine (0.03 mL under the stirring at room; 0.206 mmol), drip the key intermediate glycosyl Acibenzolar that is dissolved in dioxane behind 10 min again7b(by 90% purity meter, 60 mg, 0.0989 mmol), a complete back room temperature continues to stir 6 h, and the ESI-MS detection reaction is complete, so with getting bullion behind the reaction solution cryoconcentration9b, directly carry out next step reaction.

(3) preparation midbody (9c)

With commercially available Caspofungin acetate8(50 mg; 0.0412 mmol) place 25 mL eggplant-shape bottles, add each 2 mL of entry and dioxane and make it to dissolve fully, drip triethylamine (0.03 mL under the stirring at room; 0.206 mmol), drip the key intermediate glycosyl Acibenzolar that is dissolved in dioxane behind 10 min again7c(by 90% purity meter, 48 mg, 0.0989 mmol), a complete back room temperature continues to stir 6 h, and the ESI-MS detection reaction is complete, so with getting bullion behind the reaction solution cryoconcentration9c, directly carry out next step reaction.

(4) preparation midbody (9d)

With commercially available Caspofungin acetate8(50 mg; 0.0412 mmol) place 25 mL eggplant-shape bottles, add each 2 mL of entry and dioxane and make it to dissolve fully, drip triethylamine (0.03 mL under the stirring at room; 0.206 mmol), drip the key intermediate glycosyl Acibenzolar that is dissolved in dioxane behind 10 min again7d(by 90% purity meter, 50 mg, 0.0989 mmol), a complete back room temperature continues to stir 6 h, and the ESI-MS detection reaction is complete, so with getting bullion behind the reaction solution cryoconcentration9d, directly carry out next step reaction.

(5) preparation midbody (9e)

With commercially available Caspofungin acetate8(50 mg; 0.0412 mmol) place 25 mL eggplant-shape bottles, add each 2 mL of entry and dioxane and make it to dissolve fully, drip triethylamine (0.03 mL under the stirring at room; 0.206 mmol), drip the key intermediate glycosyl Acibenzolar that is dissolved in dioxane behind 10 min again7e(by 90% purity meter, 84 mg, 0.0989 mmol), a complete back room temperature continues to stir 6 h, and the ESI-MS detection reaction is complete, so with getting bullion behind the reaction solution cryoconcentration9e, directly carry out next step reaction.

(6) preparation midbody (9f)

With commercially available Caspofungin acetate8(50 mg; 0.0412 mmol) place 25 mL eggplant-shape bottles, add each 2 mL of entry and dioxane and make it to dissolve fully, drip triethylamine (0.03 mL under the stirring at room; 0.206 mmol), drip the key intermediate glycosyl Acibenzolar that is dissolved in dioxane behind 10 min again7f(by 90% purity meter, 84 mg, 0.0989 mmol), a complete back room temperature continues to stir 6 h, and the ESI-MS detection reaction is complete, so with getting bullion behind the reaction solution cryoconcentration9f, directly carry out next step reaction.

Seven, preparation target compound (1a-f)

(1) preparation target compound (1a), i.e. compound 1a in the table 1

With step 6 gained bullion9aPlace 25 mL eggplant-shape bottles, add the CH of pH=8₃ONa/CH₃OH solution, stirred overnight at room temperature, next day, ESI-MS detected; React complete, so transfer pH=7 with acetic acid, concentrate under reduced pressure at low temperature; Gained oily matter is crossed preparation HPLC purifying after revolving acetic acid with the toluene band; Moving phase is acetonitrile and water (0.1% trifluoroacetic acid, v/v=20:80 ~ 80:20), get white powder after the lyophilize1a, weighing 50 mg, two step total recoverys are 73 %.

(2) preparation target compound (1b), i.e. compound 1b in the table 1

With step 6 gained bullion9bPlace 25 mL eggplant-shape bottles, add the CH of pH=8₃ONa/CH₃OH solution, stirred overnight at room temperature, next day, ESI-MS detected; React complete, so transfer pH=7 with acetic acid, concentrate under reduced pressure at low temperature; Gained oily matter is crossed preparation HPLC purifying after revolving acetic acid with the toluene band; Moving phase is acetonitrile and water (0.1% trifluoroacetic acid, v/v=20:80 ~ 80:20), get white powder after the lyophilize1b, weighing 51 mg, two step total recoverys are 75 %.

(3) preparation target compound (1c), i.e. compound 1c in the table 1

With step 6 gained bullion9cPlace 25 mL eggplant-shape bottles, add the CH of pH=8₃ONa/CH₃OH solution, stirred overnight at room temperature, next day, ESI-MS detected; React complete, so transfer pH=7 with acetic acid, concentrate under reduced pressure at low temperature; Gained oily matter is crossed preparation HPLC purifying after revolving acetic acid with the toluene band; Moving phase is acetonitrile and water (0.1% trifluoroacetic acid, v/v=20:80 ~ 80:20), get white powder after the lyophilize1c, weighing 46 mg, two step total recoverys are 71 %.

(4) preparation target compound (1d), i.e. compound 1d in the table 1

With step 6 gained bullion9dPlace 25 mL eggplant-shape bottles, add the CH of pH=8₃ONa/CH₃OH solution, stirred overnight at room temperature, next day, ESI-MS detected; React complete, so transfer pH=7 with acetic acid, concentrate under reduced pressure at low temperature; Gained oily matter is crossed preparation HPLC purifying after revolving acetic acid with the toluene band; Moving phase is acetonitrile and water (0.1% trifluoroacetic acid, v/v=20:80 ~ 80:20), get white powder after the lyophilize1d, weighing 49 mg, two step total recoverys are 74 %.

(5) preparation target compound (1e), i.e. compound 1e in the table 1

With step 6 gained bullion9ePlace 25 mL eggplant-shape bottles, add the CH of pH=8₃ONa/CH₃OH solution, stirred overnight at room temperature, next day, ESI-MS detected; React complete, so transfer pH=7 with acetic acid, concentrate under reduced pressure at low temperature; Gained oily matter is crossed preparation HPLC purifying after revolving acetic acid with the toluene band; Moving phase is acetonitrile and water (0.1% trifluoroacetic acid, v/v=20:80 ~ 80:20), get white powder after the lyophilize1e, weighing 57 mg, two step total recoverys are 71 %.

(6) preparation target compound (1f), i.e. compound 1f in the table 1

With step 6 gained bullion9fPlace 25 mL eggplant-shape bottles, add the CH of pH=8₃ONa/CH₃OH solution, stirred overnight at room temperature, next day, ESI-MS detected; React complete, so transfer pH=7 with acetic acid, concentrate under reduced pressure at low temperature; Gained oily matter is crossed preparation HPLC purifying after revolving acetic acid with the toluene band; Moving phase is acetonitrile and water (0.1% trifluoroacetic acid, v/v=20:80 ~ 80:20), get white powder after the lyophilize1f, weighing 58 mg, two step total recoverys are 71 %.

Need to prove that when all the other target compounds prepare, like n=1 or, adopt the sugar of corresponding R group at 2 o'clock, corresponding side chain is as raw material, method is the same.Agents useful for same is commercially available analytical pure among the embodiment, and preparation liquid phase solvent for use is a chromatographically pure, all purchases the chemical reagents corporation in Shanghai.

The pharmacological evaluation of embodiment 3 The compounds of this invention

(1) experimental technique: adopt Association for Standardization of U.S. clinical labororatory (Clinical and Laboratory Standards Institute, CLSI) CLSI-M27A3 and the M38-A2 file micro-liquid base dilution method of being recommended.

(1) experimental strain

This experiment has selected for use the common human body cause illness's standard fungal bacterial strain of following 6 kind of 7 strain as screening object:

Two strain ATCC type strains:

Candida albicans (Candida albicans) SC5314

Cryptococcus neoformans (Cryptococcus neoformans) 32609

Five strain clinical strains:

The Candida glabrata bacterium (Candida krusei) 537

Candida parapsilosis (Candida parapsilosis) 22019

Trichophyton (Trichophyton rubrum) Cmccftla

Gypsum shape sporidiole bacteria (Microsporum gypseum) Cmccfmza

Candida albicans (Candida albicans) Y0109

(2) TP

The bacteria suspension preparation: above-mentioned candidiasis was cultivated 24 hours for 35 ℃ through the SDA substratum, and filamentous fungus was through PDA substratum activation 7 days, and twice activation is with the blood cell counting plate counting, with RPMI 1640 liquid nutrient mediums adjustment bacteria concentration to 1 * 10³-2 * 10⁴Individual/mL.

Soup preparation: get testing compound of the present invention and be dissolved in methyl-sulphoxide, be made into the medicine storage liquid of 6.4 mmol/L ,-70 ℃ of preservations.Be diluted to 640 μ mol/L with RPMI 1640 substratum before the experiment.

Inoculation: No. 1 hole of 96 orifice plates adds RPMI 1,640 200 μ L and makes blank; The 3-12 hole respectively adds RPMI 1,640 100 μ L; No. 2 the hole adds RPMI 1,640 180 μ L and 640 μ mol/L soups, 20 μ L, 10 grades of doubling dilutions of the drug level in 2-11 hole, and each hole drug level is respectively 64,32,16,8,4,2,1,0.5,0.25,0.125 μ mol/L; No. 12 the hole does not add soup, makes positive control.The medicine contrast is fluconazole (FCZ.).The bacteria suspension for preparing is seeded to the 2nd ~ No. 12 hole of 96 orifice plates with multichannel pipettor with 100 μ L bacterium liquid, and final inoculum density is 0.5 * 10³-2.5 * 10³CFU/mL; Last row of 96 orifice plates is Quality Control bacterial strain Candida parapsilosis ATCC22019.

Cultivate and detect: 96 orifice plates that will inoculate leave standstill cultivation after 1-7 days, observations in 35 ℃.If positive control hole OD value (OD value) is 100%, being lower than 80% lowest drug concentration with OD value than positive control hole is minimal inhibitory concentration value (MIC₈₀).

(2) experimental result

External bacteriostatic experiment result sees table 2.

Table 2 compound of the present invention1a-fExternal antimycotic experimental result

Annotate: CAS is contrast medicine Caspofungin.

Visible by last table, The compounds of this invention has the selected fungi of major part and suppresses active, though the glycosyl derivatives molecular weight increases, it is active to the vitro inhibition of part bacterial strain that institute's synthetic target compound has kept basically, some compound as1a,1bThe isoreactivity comparison is according to medicine Caspofungin better effects if, so The compounds of this invention can be used for preparing new antifungal drug.

The stability experiment of embodiment 4 The compounds of this invention

The improvement situation of the relative Caspofungin stability of synthetic glycosyl derivatives is chosen target compound in order to investigate1aWith1e, and it has been carried out high humidity and thimble test.

(1) experimental technique

High wet test: in a moisture eliminator, place saturated sodium nitrite solution, keep its interior relative humidity, will fill compound then 75%1a,1eWith contrast medicine Caspofungin1Sample bottle place wherein, room temperature was observed 10 days.Pick and place the sample of having put 0 day, 5 days and 10 days respectively and with performance liquid HPLC it is carried out content analysis through analyzing.

High temperature test: will fill compound1a,1eWith contrast medicine Caspofungin1Sample bottle be positioned over a temperature maintenance in 60 ℃ thermostat container, observed 10 days.Pick and place the sample of having put 0 day, 5 days and 10 days respectively and with performance liquid HPLC it is carried out content analysis through analyzing.

(2) experimental result

The stability experiment result sees Fig. 2.From figure2ACan find out compound1a,1eWith contrast medicine Caspofungin1After 5 days, HPLC analyzes demonstration, compound in this super-humid conditions held1aWith1eAlmost not influence of content, and contrast medicine Caspofungin1Content but descended about 2%; After 10 days, compound1aWith1eContent descended about 0.5% and 3.5% respectively, and contrast medicine Caspofungin1Content but descended approximately 6.5%, be respectively compound1aWith1e13 times and 2 times.Above result shows that the glycosyl derivatives of Caspofungin is than guide's thing Caspofungin1Be difficult for the moisture absorption.From figure2BCan find out compound1a,1eWith contrast medicine Caspofungin1All unstable under this hot conditions, but compound1aWith1eThan guide's thing Caspofungin1Stable, for example, in this hot conditions held after 10 days, compound1aWith1eContent descended about 40%, and contrast medicine Caspofungin1Content but descended up to 70% more than.

The formulation preparation of embodiment 10 The compounds of this invention

One, prepare tablet according to methods known in the art, every contains following compositions: compound1a50mg, lactose 70mg, Magnesium Stearate 3mg, Vinylpyrrolidone polymer 7mg adds up to 130mg.

Two,Prepare tablet according to methods known in the art, every contains following compositions: compound1e50mg, lactose 70mg, Magnesium Stearate 3mg, Vinylpyrrolidone polymer 7mg adds up to 130mg.

Three,Prepare capsule according to methods known in the art, contain following compositions in each capsule: compound1a50mg, lactose 70mg, W-Gum 25mg, Magnesium Stearate 1mg, Vinylpyrrolidone polymer 4mg adds up to 150mg.

Four,Prepare capsule according to methods known in the art, contain following compositions in each capsule: compound1e50mg, lactose 70mg, W-Gum 25mg, Magnesium Stearate 1mg, Vinylpyrrolidone polymer 4mg adds up to 150mg.

The above only is a preferred implementation of the present invention; Should be pointed out that for those skilled in the art, under the prerequisite that does not break away from the inventive method; Can also make some improvement and replenish, these improvement and replenish and also should be regarded as protection scope of the present invention.

Claims (7)

Translated fromChinese

1.一种糖肽类抗真菌化合物及其药学上可接受的盐，其特征在于，所述的糖肽类抗真菌化合物结构通式如下：1. A glycopeptide antifungal compound and a pharmaceutically acceptable salt thereof, characterized in that, the general structural formula of the glycopeptide antifungal compound is as follows:

其中，n等于1，2或3；where n is equal to 1, 2 or 3;R为单糖或二糖；R is monosaccharide or disaccharide;所述的单糖为葡萄糖、半乳糖、木糖、鼠李糖、甘露糖、核糖、氨基葡萄糖或乙酰氨基葡萄糖；The monosaccharide is glucose, galactose, xylose, rhamnose, mannose, ribose, glucosamine or acetylglucosamine;所述的二糖为麦芽糖或乳糖。Described disaccharide is maltose or lactose.2.根据权利要求1所述的糖肽类抗真菌化合物及其药学上可接受的盐，其特征在于，所述的n等于3。2. The glycopeptide antifungal compound and the pharmaceutically acceptable salt thereof according to claim 1, wherein said n is equal to 3.3.根据权利要求2所述的糖肽类抗真菌化合物及其药学上可接受的盐，其特征在于，所述的R为葡萄糖或麦芽糖。3. The glycopeptide antifungal compound and the pharmaceutically acceptable salt thereof according to claim 2, wherein said R is glucose or maltose.4.根据权利要求1所述的糖肽类抗真菌化合物及其药学上可接受的盐，其特征在于，所述的药学上可接受的盐类是盐酸盐、硫酸盐、硫酸氢盐、氢溴酸盐、醋酸盐、草酸盐、柠檬酸盐或甲磺酸盐。4. Glycopeptide antifungal compounds and pharmaceutically acceptable salts thereof according to claim 1, wherein the pharmaceutically acceptable salts are hydrochloride, sulfate, bisulfate, Hydrobromide, Acetate, Oxalate, Citrate or Methanesulfonate.5.一种药物组合物，其特征在于，它含有权利要求1-4任一所述的糖肽类抗真菌化合物及其药学上可接受的盐，并含有常规药用载体。5. A pharmaceutical composition, characterized in that it contains the glycopeptide antifungal compound and its pharmaceutically acceptable salt according to any one of claims 1-4, and contains conventional pharmaceutical carriers.6.一种如权利要求1-4任一所述的糖肽类抗真菌化合物及其药学上可接受的盐的制备方法，其特征在于，该方法包括下列步骤：6. A method for preparing a glycopeptide antifungal compound and a pharmaceutically acceptable salt thereof according to any one of claims 1-4, characterized in that the method comprises the following steps:a）制备关键中间体糖基活化酯：以糖基为起始原料，与醋酸钠在醋酸酐中回流反应得到全乙酰保护的糖基；然后在三甲基叠氮硅烷和三氟化硼乙醚存在的条件下发生取代反应，将1-位的乙酰氧基变为叠氮基；叠氮基通过10% Pd-C催化氢化生成氨基后在4-(N,N-二甲氨基)吡啶存在的条件下与相应的酸酐缩合成酰胺；裸露出来的羧基在与N,N-二环己基碳二亚胺存在的条件下与N-羟基丁二酰亚胺发生酯缩合生成相应的糖基活化酯；a) Preparation of the key intermediate glycosyl activated ester: using glycosyl as the starting material, reflux reaction with sodium acetate in acetic anhydride to obtain a fully acetyl-protected glycosyl; then trimethylazidesilane and boron trifluoride ether Under the conditions of existence, a substitution reaction occurs, and the acetoxy group at the 1-position is changed to an azido group; the azido group is catalyzed by 10% Pd-C to generate an amino group and exists in 4-(N,N-dimethylamino)pyridine Condensation with the corresponding acid anhydride to form an amide under certain conditions; the exposed carboxyl group undergoes ester condensation with N-hydroxysuccinimide in the presence of N,N-dicyclohexylcarbodiimide to generate the corresponding glycosyl activation ester;b）制备目标化合物Caspofungin糖基衍生物：步骤a）制备的关键中间体糖基活化酯与Caspofungin醋酸盐在三乙胺碱性条件下发生缩合生成全乙酰保护的Caspofungin糖基化合物，然后在甲醇钠/甲醇溶液中脱去乙酰基保护即得目标化合物Caspofungin糖基衍生物。b) Preparation of the target compound Caspofungin glycosyl derivatives: step a) The key intermediate glycosyl activated ester prepared and Caspofungin acetate is condensed under triethylamine alkaline conditions to generate a fully acetyl-protected Caspofungin glycosyl compound, and then in Deprotection of the acetyl group in sodium methoxide/methanol solution to obtain the target compound Caspofungin glycosyl derivative.7.如权利要求1-4任一所述的糖肽类抗真菌化合物及其药学上可接受的盐在制备治疗真菌感染药物中的应用。7. The use of the glycopeptide antifungal compound according to any one of claims 1-4 and a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating fungal infections.

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