CN111662351B - New octreolone type sapogenin derivative and application thereof in preparation of drug-resistant bacteria resistant drugs - Google Patents

New octreolone type sapogenin derivative and application thereof in preparation of drug-resistant bacteria resistant drugs Download PDF

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CN111662351B
CN111662351B CN202010548873.1A CN202010548873A CN111662351B CN 111662351 B CN111662351 B CN 111662351B CN 202010548873 A CN202010548873 A CN 202010548873A CN 111662351 B CN111662351 B CN 111662351B
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CN111662351A (en
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毕毅
曹玉成
王恺奕
孟庆国
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Yantai University
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    • C07J—STEROIDS
    • C07J17/00—Normal steroids containing carbon, hydrogen, halogen or oxygen, having an oxygen-containing hetero ring not condensed with the cyclopenta(a)hydrophenanthrene skeleton
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
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    • Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Abstract

The invention discloses a new ocrtalone type sapogenin derivative and application thereof in preparing a drug-resistant bacterium resistant drug. The preparation method of the oxtriptolone type ginsengenin derivative comprises the following steps: a.20 (ii) (S) -protopanaxadiol is used as a raw material, and acetic anhydride protects hydroxyl at C-3 and C-12 positions; b. oxidizing m-chloroperoxybenzoic acid to ensure that a tetrahydrofuran ring is formed at the C-24 position; c. removing the protecting groups at C-3 and C-12 positions by sodium hydroxide; d. under the catalysis of DMAP and EDCI, reacting with different Boc-amino acid and Fmoc-Boc-amino acid; e. removing Boc protecting group under the catalysis of trifluoroacetic acid to obtain a crude product, and purifying by column chromatography to obtain a target compound; f. reacting amino with dansyl chloride under the catalysis of DIEA; g. removing Fmoc protection under piperidine catalysis to obtain a crude product, and purifying by column chromatography to obtain the target compound. The oxtriptolone type ginsenoside of the inventionThe derivative has good antibacterial effect, and has remarkably improved inhibition effect on methicillin-resistant Staphylococcus aureus compared with the oxtriptolone type ginsenoside mother nucleus.

Description

New octreolone type sapogenin derivative and application thereof in preparation of drug-resistant bacteria resistant drugs
Technical Field
The invention relates to the field of organic synthesis and pharmaceutical chemistry, in particular to an oxtriptolone type ginsenoside derivative with a novel structure. The invention discloses a preparation method of the oxtriptolone type ginsenoside derivatives, a pharmaceutical composition containing the oxtriptolone type ginsenoside derivatives and application of the oxtriptolone type ginsenoside derivatives in resisting bacterial infection diseases.
Background
The natural product is an important source of lead compounds in medicine research and development, has the characteristics of rich structure types, small toxic and side effects and the like, and therefore, the search of components with antibacterial effects from the natural product becomes a hot direction for research of scientific researchers. The ocotillol-type ginsenoside is dammarane type tetracyclic triterpene saponin with tetrahydrofuran ring on side chain, mainly exists in Panax plant, and has antitumor effect.
The discovery and research of antibacterial drugs play an important role in the treatment of infectious diseases caused by bacteria. However, abuse of antibacterial drugs has prompted bacterial resistance. Staphylococcus aureus is one of the most common pathogenic bacteria of hospital and community acquired infections. In recent years, the development of drug resistant staphylococcus aureus strains, such as methicillin-resistant staphylococcus aureus (MRSA), has been caused by the large use of antibiotics in clinical practice. MRSA, commonly referred to as superbacteria, cause infections with high morbidity and mortality, and are clinically difficult to treat. The development of new highly effective drug resistant bacteria drugs (methicillin-resistant staphylococcus aureus (MRSA)) is therefore of great importance for combating increasingly serious bacterial infections.
Disclosure of Invention
The invention provides a new oxtriptolone type sapogenin derivative which has better drug-resistant antibacterial activity, and also provides a preparation method and application of the derivative.
The invention aims to solve the technical problem of searching a new structural type compound with excellent antibacterial activity and further providing an application of the compound in preparing a methicillin-resistant staphylococcus aureus antibacterial drug.
In order to solve the technical problems, the invention provides the following technical scheme:
oxtriptolone type ginsenoside derivative shown in general formula (I) and pharmaceutically acceptable salt thereof,
Figure BDA0002541728210000011
wherein the content of the first and second substances,
R 1 represents-NH 2 、-R 3 NH 2 、
Figure BDA0002541728210000021
-CH 3 、H、
Figure BDA0002541728210000022
R 2 Represents
Figure BDA0002541728210000023
R 3 Represents (C1-C8) straight-chain or branched alkyl;
R 4 represents (C1-C3), (C5-C8) straight chain or branched chain alkyl.
Preferably, the compounds and their pharmaceutically acceptable salts, wherein,
R 1 represents-NH 2 、-R 3 NH 2 、
Figure BDA0002541728210000024
-CH 3 、H、
Figure BDA0002541728210000025
R 2 Represents
Figure BDA0002541728210000026
R 3 Represents (C1-C8) straight-chain or branched alkyl;
R 4 represents (C1-C3), (C5-C8) straight chain or branched chain alkyl.
Preferably, some of the compounds of the present invention are:
(20S, 24R) -epoxy-3 β -oxy- [2- (N' -Fmoc) -5-aminobutyryl ] -dammarane-12 β, 25-diol;
(20S, 24S) -epoxy-3 β -oxy- [2- (N' -Fmoc) -5-aminobutyryl ] -dammarane-12 β, 25-diol;
(20S, 24R) -epoxy-3 β -oxy- [2- (N' -Fmoc) -6-aminopentanoyl ] -dammarane-12 β, 25-diol;
(20S, 24S) -epoxy-3 β -oxy- [2- (N' -Fmoc) -6-aminopentanoyl ] -dammarane-12 β, 25-diol;
(20S, 24R) -epoxy-3 β -oxy- [ 2-amino-5- (5-dimethylamino-1-naphthalenesulfonamide) butyryl ] -dammarane-12 β, 25-diol;
(20s, 24s) -epoxy-3 β -oxy- [ 2-amino-5- (5-dimethylamino-1-naphthalenesulfonamide) butyryl ] -dammarane-12 β, 25-diol;
(20S, 24R) -epoxy-3 β -oxy- [ 2-amino-6- (5-dimethylamino-1-naphthalenesulfonamide) pentanoyl ] -dammarane-12 β, 25-diol;
(20S, 24S) -epoxy-3 β -oxy- [ 2-amino-6- (5-dimethylamino-1-naphthalenesulfonamide) pentanoyl ] -dammarane-12 β, 25-diol;
(20S, 24R) -epoxy-3 β -oxy- [2- (5-dimethylamino-1-naphthalenesulfonamide) -6-aminocaproyl ] -dammarane-12 β, 25-diol;
(20S, 24S) -epoxy-3 beta-oxy- [2- (5-dimethylamino-1-naphthalenesulfonamide) -6-aminocaproyl ] -dammarane-12 beta, 25-diol.
The preparation method of the oxtriptolone type ginsenoside derivative comprises the following steps:
a.20 (S) -protopanaxadiol is taken as a raw material, and acetic anhydride protects hydroxyl at C-3 and C-12 positions;
b. oxidizing m-chloroperoxybenzoic acid to ensure that a tetrahydrofuran ring is formed at the C-24 position;
c. removing the protecting groups at C-3 and C-12 positions by sodium hydroxide;
d. reacting with different Boc-amino acids and Fmoc-Boc-amino acids under the catalysis of DMAP and EDCI;
e. removing Boc protecting group under the catalysis of trifluoroacetic acid to obtain a crude product, and purifying by column chromatography to obtain a target compound;
f. reacting amino with dansyl chloride under the catalysis of DIEA;
g. removing Fmoc protection under piperidine catalysis to obtain a crude product, and purifying by column chromatography to obtain the target compound.
The invention comprises a compound of formula (I) or a salt thereof and a pharmaceutically acceptable carrier for the manufacture of a medicament for the treatment of diseases or conditions in mammals, preferably humans, including sepsis, pneumonia, meningitis, endocarditis, bone and joint infections, burn infections, surgical and wound infections, skin infections, and the like.
The application of the oxtriptolone type ginsenoside derivative shown in the general formula (I) and the medically acceptable salt thereof in preparing antibacterial drugs.
Preferably, the oxtriptolone type ginsenoside derivative shown in the general formula (I) and the medically acceptable salt thereof are applied to the preparation of the methicillin-resistant staphylococcus aureus antibacterial drug.
Drug-resistant staphylococcus aureus strains such as methicillin-resistant staphylococcus aureus (MRSA) have drug resistance to beta-lactam antibiotics, and are resistant to most of aminoglycosides, fluoroquinolones, macrolides and other antibacterial drugs, and even vancomycin-resistant MRSA strains have appeared in some areas. However, the antibacterial activity result of the invention shows that the inhibition effect of the oxtripteryl-type ginsengenin derivative obtained by structural modification and reformation on methicillin-resistant staphylococcus aureus is obviously enhanced. The oxtriptolone type ginsenoside derivative has a good antibacterial effect, and compared with an oxtriptolone type ginsenoside parent nucleus, the oxtriptolone type ginsenoside derivative disclosed by the invention has the advantages that the examples 1-10 shown in the invention have good activity on methicillin-resistant staphylococcus aureus, and the MIC value is mainly distributed in the range of 1-16 mu g/mL, so that the oxtriptolone type ginsenoside derivative can be used for preparing antibacterial infection medicines. Therefore, the derivative disclosed by the invention is a novel oxtriptolone type ginsenoside derivative with methicillin-resistant staphylococcus aureus antibacterial activity.
Therefore, the invention provides the oxtriptolone type ginsengenin derivative which has better drug-resistant antibacterial activity and a preparation method of the derivative. Compared with an ocreten-type ginsenoside mother nucleus, the ocreten-type ginsenoside derivative has better antibacterial activity on methicillin-resistant staphylococcus aureus, and the MIC value is within the range of 1-16 mu g/mL. The invention provides application of a medicament for preparing methicillin-resistant staphylococcus aureus (MRSA) infection diseases or symptoms.
Detailed Description
The present invention will be described in further detail below by way of examples, but the present invention is not limited to only the following examples.
Example 1
(20S, 24R) -epoxy-3 beta-oxy- [2- (N' -Fmoc) -5-aminobutyryl ] -dammarane-12 beta, 25-diol
20 (S) -Protopanaxadiol (500.0mg, 1.1mmol) was dissolved in chloroform (3.0 mL), DMAP (20.0mg, 0.2mmol) was added thereto, and the mixture was stirred, then acetic anhydride (0.4mL, 4.4mmol) was slowly added dropwise, and the mixture was stirred at room temperature for 1 hour. Diluting with ethyl acetate (20.0 mL), washing with 10% hydrochloric acid until acidic, washing with water, washing with saturated brine, drying over anhydrous sodium sulfate, filtering, concentrating, and performing column chromatography (petroleum ether: ethyl acetate =10: 1) to obtain 20 (S) -3, 12-diacetyl protopanaxadiol (500.0 mg, 85%) as a white solid.
20 (S) -3, 12-diacetyl protopanaxadiol (210.0mg, 0.4mmol) was dissolved in anhydrous dichloromethane (6.0 mL), and a solution of m-chloroperoxybenzoic acid (190.0mg, 0.2mmol, 75%) in dichloromethane (5.0 mL) was slowly added dropwise with cooling in an ice salt bath, after 0.5h, the mixture was warmed to room temperature and stirred for 2h. Isopropanol (1.0 mL) was added, stirring was continued for 1h, a saturated sodium bicarbonate solution was added, stirring was continued for 1h, followed by liquid-separation extraction, and the organic phase was washed successively with a saturated sodium thiosulfate solution, water, and a saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated, and subjected to column chromatography (petroleum ether: ethyl acetate =8: 1) to give 20 (S) -20, 24-epoxy-3, 12-diacetyl protopanaxadiol (146.0mg, 70.0%) as a white solid.
20 (S) -20, 24-epoxy-3, 12-diacetyl protopanaxadiol (130.0mg, 0.2mmol) was dissolved in methanol (8.0 mL), and potassium hydroxide (85.0mg, 1.5mmol) was added thereto, followed by stirring at 135 ℃ for reaction for 2 hours. After the reaction solution was cooled to room temperature, an appropriate amount of water was added, and a large amount of white solid was precipitated, which was then subjected to suction filtration, drying, and column chromatography (petroleum ether: ethyl acetate =2: 1-1.
OR (80.0mg, 0.2mmol) was dissolved in anhydrous dichloromethane (5.0 mL), and 4-dimethylaminopyridine (56.0mg, 0.5mmol), 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (96.0mg, 0.5mmol) and (R) -4- (Boc-amino) -2- (Fmoc-amino) butyric acid (110.0mg, 0.3mmol) were added to react at room temperature for 3 hours. The reaction solution was diluted with dichloromethane, washed with 5% hydrochloric acid, washed with deionized water, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated. The dried intermediate was dissolved in anhydrous dichloromethane, and excess trifluoroacetic acid was added to react at room temperature for 1h. The reaction solution was distilled under reduced pressure and subjected to silica gel column chromatography (chloroform: methanol =80:1 to 50. 1 H-NMR(CDCl 3 ,400MHz)δ(ppm):8.27(s,2H,-NH 2 ),7.70(d,J＝7.6Hz,2H,Ar-H×2),7.55(d,J＝6.6Hz,2H,Ar-H×2),7.34(t,J＝7.4Hz,2H,Ar-H×2),7.26(t,J＝7.5Hz,2H,Ar-H×2),6.20(d,J＝7.4Hz,1H,-NH-),4.47(d,J＝7.3Hz,1H,-OCH-),4.32(d,J＝6.5Hz,3H,-OCH 2 -,-NCH-),4.14(t,J＝6.8Hz,1H,-CH-),3.81(t,J＝7.6Hz,1H,-OCH-),3.60–3.47(m,1H,-OCH-),3.04(d,J＝43.0Hz,2H,-NCH 2 -),2.23(d,J＝55.9Hz,2H,-CH 2 -),1.26(s,3H,-CH 3 ),1.24(s,3H,-CH 3 ),1.22(s,3H,-CH 3 ),1.09(s,3H,-CH 3 ),0.94(s,3H,-CH 3 ),0.86(s,3H,-CH 3 ),0.82(s,3H,-CH 3 ),0.79(s,3H,-CH 3 ).
Example 2
(20S, 24S) -epoxy-3 beta-oxy- [2- (N' -Fmoc) -5-aminobutyryl group]Dammarane-12 β, 25-diol OS (80.0 mg,0.2 mmol) was dissolved in anhydrous dichloromethane (5.0 mL), and 4-dimethylaminopyridine (56.0 mg,0.5 mmol), 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (96.0 mg,0.5 mmol) and (R) -4- (Boc-amino) -2- (Fmoc-amino) butyric acid (110.0 g,0.3 mmol) were added and reacted at room temperature for 3h. The reaction solution was diluted with dichloromethane, washed with 5% hydrochloric acid, washed with deionized water, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated. The dried intermediate was dissolved in anhydrous dichloromethane, and excess trifluoroacetic acid was added to react at room temperature for 1h.The reaction solution was distilled under reduced pressure and then subjected to silica gel column chromatography (chloroform: methanol =80: 1-50) to obtain a white solid (94.3mg, 70.1%). 1 H-NMR(CDCl 3 ,400MHz)δ(ppm):8.24(s,2H,-NH 2 ),7.69(d,J＝7.5Hz,2H,Ar-H×2),7.54(d,J＝5.6Hz,2H,Ar-H×2),7.34(t,J＝7.4Hz,2H,Ar-H×2),7.25(t,J＝7.4Hz,3H,Ar-H×2),6.13(d,J＝7.5Hz,1H,-NH-),4.55–4.43(m,1H,-OCH-),4.40–4.26(m,3H,-OCH 2 -,-NCH-),4.13(t,J＝6.9Hz,1H,-CH-),3.92–3.79(m,1H,-OCH-),3.50(s,1H,-OCH-),3.02(d,J＝55.7Hz,2H,-NCH 2 -),2.26(d,J＝41.9Hz,2H,-CH 2 -),1.24(s,3H,-CH 3 ),1.23(s,3H,-CH 3 ),1.18(s,3H,-CH 3 ),1.10(s,3H,-CH 3 ),0.96(s,3H,-CH 3 ),0.85(s,3H,-CH 3 ),0.84(s,3H,-CH 3 ),0.79(s,3H,-CH 3 ).
Example 3
(20S, 24R) -epoxy-3 beta-oxy- [2- (N' -Fmoc) -6-aminopentanoyl ] -dammarane-12 beta, 25-diol
Reference (20S, 24R) -epoxy-3 beta-oxy- [2- (N' -Fmoc) -5-aminobutyryl]Process for the synthesis of (E) -dammarane-12 β, 25-diol, OR reacted with N-Fmoc-N' -Boc-L-ornithine to remove the Boc protecting group to give a white solid (95.1mg, 69.9%). 1 H-NMR(CDCl 3 ,400MHz)δ(ppm):8.01(s,2H,-NH 2 ),7.69(d,J＝7.5Hz,2H,Ar-H×2),7.54(t,J＝7.0Hz,2H,Ar-H×2),7.32(t,J＝7.5Hz,2H,Ar-H×2),7.24(d,J＝5.7Hz,2H,Ar-H×2),5.84(d,J＝8.0Hz,1H,-NH-),4.50–4.38(m,1H,-OCH-),4.27(d,J＝6.8Hz,3H,-OCH 2 -,-NCH-),4.12(t,J＝7.0Hz,1H,-CH-),3.86–3.77(m,1H,-OCH-),3.48(dd,J＝10.2,4.1Hz,1H,-OCH-),2.95(s,2H,-NCH 2 -),1.25(s,3H,-CH 3 ),1.24(s,6H,-CH 3 ×2),1.08(s,3H,-CH 3 ),0.93(s,3H,-CH 3 ),0.83(s,3H,-CH 3 ),0.79(s,3H,-CH 3 ),0.75(s,3H,-CH 3 ).
Example 4
(20S, 24S) -epoxy-3 beta-oxy- [2- (N' -Fmoc) -6-aminopentanoyl ] -dammarane-12 beta, 25-diol
Reference (20S, 24S) -epoxy-3 beta-oxy- [2- (N' -Fmoc) -5-aminobutyryl]Of (E) -dammarane-12 beta, 25-diolSynthesis procedure, OS reacted with N-Fmoc-N' -Boc-L-ornithine to remove the Boc protecting group to give a white solid (92.0 mg, 67.8%). 1 H-NMR(CDCl 3 ,400MHz)δ(ppm):8.07(s,2H,-NH 2 ),7.69(d,J＝7.5Hz,2H,Ar-H×2),7.55(t,J＝6.3Hz,2H,Ar-H×2),7.33(t,J＝7.5Hz,2H,Ar-H×2),7.24(t,J＝7.4Hz,2H,Ar-H×2),5.83(d,J＝8.0Hz,1H,-NH-),4.47(t,J＝8.7Hz,1H,-OCH-),4.28(d,J＝6.6Hz,3H,-OCH 2 -,-NCH-),4.13(t,J＝7.1Hz,1H,-CH-),3.90–3.81(m,1H,-OCH-),3.54–3.44(m,1H,-OCH-),2.97(s,2H,-NCH 2 -),1.24(s,3H,-CH 3 ),1.19(s,3H,-CH 3 ),1.09(s,3H,-CH 3 ),0.96(s,3H,-CH 3 ),0.85(s,3H,-CH 3 ),0.84(s,3H,-CH 3 ),0.77(s,3H,-CH 3 ),0.76(s,3H,-CH 3 ).
Example 5
(20S, 24R) -epoxy-3 beta-oxy- [ 2-amino- (5-dimethylamino-1-naphthalenesulfonamide) butyryl ] -dammarane-12 beta, 25-diol
The objective compound (20S, 24R) -epoxy-3. Beta. -oxy- [2- (N' -Fmoc) -5-aminobutyryl group obtained in example 1 was used][ 10 ] dammarane-12. Beta., 25-diol (60.0mg, 0.1mmol) was dissolved in anhydrous dichloromethane (5.0 mL), and N, N-diisopropylethylamine (36.0mg, 0.3mmol) and dansyl chloride (24.0mg, 0.1mmol) were added to react at room temperature for 4 hours. And (3) diluting the reaction solution with dichloromethane, washing with 5% hydrochloric acid, washing with deionized water, washing with saturated sodium chloride, drying with anhydrous sodium sulfate, and concentrating to obtain an intermediate. The dried intermediate is dissolved in anhydrous dichloromethane, and excess piperidine is added to react for 2h at room temperature. The reaction solution was distilled under reduced pressure and subjected to silica gel column chromatography (chloroform: methanol =100: 1-50) to obtain v-5 (44.0 mg, 72.2%) as a pale green solid. 1 H-NMR(CDCl 3 ,400MHz)δ(ppm):8.50(dt,J＝8.5,1.0Hz,1H,Ar-H),8.27(d,J＝8.7Hz,1H,Ar-H),8.22(dd,J＝7.3,1.3Hz,1H,Ar-H),7.51(ddd,J＝14.7,8.6,7.4Hz,2H,Ar-H×2),7.16(dd,J＝7.6,0.7Hz,1H,Ar-H),5.60(s,1H,-OH),4.36(dd,J＝10.6,5.9Hz,1H,-OCH-),3.82(dd,J＝8.8,6.5Hz,1H,-OCH-),3.49–3.42(m,1H,-OCH-),3.30(dd,J＝9.6,3.5Hz,1H,-NH 2 CH-),3.14(dd,J＝11.7,7.4Hz,1H,-NCH 2 -),3.01–2.91(m,1H,-NCH 2 -),2.87(s,6H,-CH 3 ×2),1.25(s,3H,-CH 3 ),1.24(s,3H,-CH 3 ),1.07(s,3H,-CH 3 ),0.94(s,3H,-CH 3 ),0.86(s,3H,-CH 3 ),0.81(s,3H,-CH 3 ),0.73(s,3H,-CH 3 ),0.72(s,3H,-CH 3 ).
Example 6
(20S, 24S) -epoxy-3 beta-oxy- [ 2-amino- (5-dimethylamino-1-naphthalenesulfonamide) butyryl][ 10 ] dammarane-12 β, 25-diol the title compound (20S, 24S) -epoxy-3 β -oxy- [2- (N' -Fmoc) -5-aminobutyryl group obtained in example 2 was synthesized according to the method of example 5]Reaction of dammarane-12 β, 25-diol with dansyl chloride followed by removal of the Fmoc protecting group gave V-6 as a pale green solid (42.1mg, 69.6%). 1 H-NMR(CDCl 3 ,400MHz)δ(ppm):8.51(d,J＝8.4Hz,1H,Ar-H),8.27(d,J＝8.6Hz,1H,Ar-H),8.22(d,J＝7.3Hz,1H,Ar-H),7.56(s,1H,Ar-H),7.52–7.47(m,1H,Ar-H),7.17(d,J＝7.5Hz,1H,Ar-H),5.75(s,1H,-OH),4.38(dd,J＝10.8,5.8Hz,1H,-OCH-),3.85(dd,J＝10.9,5.3Hz,1H,-OCH-),3.50(td,J＝9.8,4.7Hz,1H,-OCH-),3.30(dd,J＝9.7,3.5Hz,1H,-NH 2 CH-),3.20–3.12(m,1H,-NCH 2 -),2.97(dd,J＝8.9,4.2Hz,1H,-NCH 2 -),2.87(s,6H,-CH 3 ×2),1.26(s,3H,-CH 3 ),1.23(s,3H,-CH 3 ),1.08(s,3H,-CH 3 ),0.98(s,3H,-CH 3 ),0.88(s,3H,-CH 3 ),0.86(s,3H,-CH 3 ),0.75(s,3H,-CH 3 ),0.74(s,3H,-CH 3 ).
Example 7
(20S, 24R) -epoxy-3 beta-oxy- [ 2-amino- (5-dimethylamino-1-naphthalenesulfonamide) valeryl ] -dammarane-12 beta, 25-diol
Referring to the synthesis method of example 5, (20S, 24R) -epoxy-3. Beta. -oxy- [2- (N' -Fmoc) -6-aminoornityl group, the objective compound obtained in example 3, was prepared]Reaction of-dammarane-12 β, 25-diol with dansyl chloride followed by removal of the Fmoc protecting group gave V-7 as a pale green solid (40.0 mg, 65.1%). 1 H-NMR(CDCl 3 ,400MHz)δ(ppm):8.50(d,J＝8.5Hz,1H,Ar-H),8.29(d,J＝8.7Hz,1H,Ar-H),8.21(dd,J＝7.3,1.3Hz,1H,Ar-H),7.55–7.50(m,1H,Ar-H),7.51–7.46(m,1H,Ar-H),7.15(d,J＝7.5Hz,1H,Ar-H),5.57(s,1H.-OH),4.47–4.39(m,1H,-OCH-),3.82(dd,J＝8.8,6.6Hz,1H,-OCH-),3.49(td,J＝10.5,4.6Hz,1H,-OCH-),3.21(dd,J＝8.6,4.1Hz,1H,-NH 2 CH-),2.99–2.91(m,1H,-NCH 2 -),2.86(s,6H,-CH 3 ×2),2.81(ddd,J＝12.7,7.5,5.4Hz,1H,-NCH 2 -),1.25(s,3H,-CH 3 ),1.24(s,3H,-CH 3 ),1.07(s,3H,-CH 3 ),0.96(s,3H,-CH 3 ),0.87(s,3H,-CH 3 ),0.84(s,3H,-CH 3 ),0.77(s,3H,-CH 3 ),0.74(s,3H,-CH 3 ).
Example 8
(20S, 24S) -epoxy-3 beta-oxy- [ 2-amino- (5-dimethylamino-1-naphthalenesulfonamide) valeryl][ 10 ] dammarane-12 β, 25-diol the title compound (20S, 24S) -epoxy-3 β -oxy- [2- (N' -Fmoc) -6-aminoornityl group obtained in example 4 was synthesized according to the method of example 5]Reaction of dammarane-12 β, 25-diol with dansyl chloride followed by removal of the Fmoc protecting group gave V-8 as a pale green solid (39.1mg, 63.4%). 1 H-NMR(CDCl 3 ,400MHz)δ(ppm):8.51(d,J＝10.6Hz,1H,Ar-H),8.29(d,J＝8.7Hz,1H,Ar-H),8.24–8.19(m,1H,Ar-H),7.58–7.52(m,1H,Ar-H),7.53–7.47(m,1H,Ar-H),7.16(d,J＝8.3Hz,1H,Ar-H),5.75(s,1H,-OH),4.44(dd,J＝9.3,7.2Hz,1H,-OCH-),3.86(dd,J＝10.8,5.4Hz,1H,-OCH-),3.51(td,J＝10.3,4.8Hz,1H,-OCH-),3.23(dd,J＝8.7,4.1Hz,1H,-NH 2 CH-),3.01–2.91(m,1H,-NCH 2 -),2.87(s,6H,-CH 3 ×2),2.80(ddd,J＝12.7,7.6,5.3Hz,1H,-NCH 2 -),1.26(s,3H,-CH 3 ),1.23(s,3H,-CH 3 ),1.08(s,3H,-CH 3 ),0.99(s,3H,-CH 3 ),0.89(s,3H,-CH 3 ),0.88(s,3H,-CH 3 ),0.79(s,3H,-CH 3 ),0.75(s,3H,-CH 3 ).
Example 9
(20S, 24R) -epoxy-3 beta-oxy- [2- (5-dimethylamino-1-naphthalenesulfonamide) -6-aminocaproyl ] -dammarane-12 beta, 25-diol
Referring to example 1, OR (80.0mg, 0.2mmol) was dissolved in anhydrous dichloromethane (5.0 mL), and 4-dimethylaminopyridine (56.0mg, 0.5mmol), 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (96.1mg, 0.5mmol) and N-Boc-N' -Fmoc-L-lysine (116.0mg, 0.3mmol) were added to react at room temperature for 3 hours. For reaction solution IIDiluting chloromethane, washing with 5% hydrochloric acid, washing with deionized water, washing with saturated sodium chloride, drying with anhydrous sodium sulfate, and concentrating to obtain yellow intermediate X1. And dissolving the dried intermediate X1 in anhydrous dichloromethane, adding excess trifluoroacetic acid (1.0 ml,13.4 mmol), reacting at room temperature for 1h, and distilling and drying the reaction solution under reduced pressure to obtain a yellow intermediate X2. The dried intermediate X2 was dissolved in anhydrous dichloromethane (5.0 ml), N-diisopropylethylamine (36.0 mg,0.3 mmol) and dansyl chloride (24.2 mg,0.1 mmol) were added, and the Fmoc protecting group was removed after reaction at room temperature for 4 hours, followed by silica gel column chromatography to give V-9 (80.1mg, 57.4%) as a pale green solid. 1 H-NMR(CDCl 3 ,400MHz)δ(ppm):8.51(d,J＝8.5Hz,1H,Ar-H),8.29(d,J＝8.7Hz,1H,Ar-H),8.21(d,J＝7.3Hz,1H,Ar-H),7.60–7.52(m,1H,Ar-H),7.52–7.44(m,1H,Ar-H),7.16(d,J＝6.8Hz,1H,Ar-H),4.15(dd,J＝11.8,4.5Hz,1H,-OCH-),3.86–3.78(m,2H,-OCH-,COCH-),3.53–3.41(m,1H,-OCH-),2.85(s,6H,-CH 3 ×2),1.26(s,3H,-CH 3 ),1.24(s,3H,-CH 3 ),1.08(s,3H,-CH 3 ),0.93(s,3H,-CH 3 ),0.83(s,3H,-CH 3 ),0.77(s,3H,-CH 3 ),0.67(s,3H,-CH 3 ),0.66(s,3H,-CH 3 ).
Example 10
(20S, 24S) -epoxy-3 beta-oxy- [2- (5-dimethylamino-1-naphthalenesulfonamide) -6-aminocaproyl ] -dammarane-12 beta, 25-diol
OS (80.0mg, 0.2mmol) was dissolved in anhydrous dichloromethane (5.0 mL), and 4-dimethylaminopyridine (56.0mg, 0.5mmol), 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (96.0mg, 0.5mmol) and N-Boc-N' -Fmoc-L-lysine (116.2mg, 0.3mmol) were added to react at room temperature for 3 hours. The reaction solution was diluted with dichloromethane, washed with 5% HCl, washed with deionized water, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated to obtain white intermediate X3. Dissolving the dried intermediate X3 in anhydrous dichloromethane, adding excessive trifluoroacetic acid (1.0 ml,13.4 mmol), reacting at room temperature for 1h, and distilling the reaction solution under reduced pressure to obtain a white intermediate X4. The dried intermediate X4 was dissolved in anhydrous dichloromethane (5.0 ml), N-diisopropylethylamine (36.1mg, 0.3mmol) and dansyl chloride (24.0mg, 0.1mmol) were added to the reaction solution to remove the Fmoc protecting group, and the mixture was subjected to silica gel column chromatography to give V-10 (80.1mg, 57.4%) as a pale green solid
1 H-NMR(CDCl 3 ,400MHz)δ(ppm):8.48(d,J＝8.5Hz,1H,Ar-H),8.32(d,J＝8.7Hz,1H,Ar-H),8.20(dd,J＝7.3,1.1Hz,1H,Ar-H),7.58–7.52(m,1H,Ar-H),7.45(dd,J＝8.5,7.4Hz,1H,Ar-H),7.14(d,J＝7.5Hz,1H,Ar-H),5.72(s,1H,-OH),4.11(dd,J＝11.8,4.4Hz,1H,-OCH-),3.90–3.77(m,2H,-OCH-,COCH-),3.46(td,J＝10.1,4.5Hz,1H,-OCH-),2.83(s,6H,-CH 3 ×2),1.24(s,3H,-CH 3 ),1.21(s,3H,-CH 3 ),1.08(s,3H,-CH 3 ),0.94(s,3H,-CH 3 ),0.83(s,3H,-CH 3 ),0.77(s,3H,-CH 3 ),0.62(s,3H,-CH 3 ),0.61(s,3H,-CH 3 ).
Pharmacological tests prove that the oxtriptolone type ginsenoside derivative has a good antibacterial effect and can be used for preparing anti-infective medicaments.
The following are the results of pharmacological experiments with some of the compounds of the invention.
1 apparatus and equipment:
water proof formula constant temperature incubator: model GNP-9080, shanghai sperm macro laboratory Equipment Co., ltd
An enzyme-labeling instrument: model 800TS, serial No. 17101018, bioTek instruments, inc
An electronic balance: model PL203, mettlerToledo group
An electronic balance: model DT1000, normal commercial weighing apparatus works
Vertical pressure steam sterilizer: model LDZX-75KBS, shanghai Shenan medical Instrument plant
2 cell lines and reagents:
dimethyl sulfoxide (DMSO)
Sodium chloride injection
MH broth (MHB)
MH (A) culture medium
Selecting bacteria: methicillin-resistant Staphylococcus aureus MRSA18-19, MRSA18-20, ATCC29213.
MH (A) culture medium is streaked, after 24 hours of incubation at 35-37 ℃, single colony is picked, and the McLeod turbidimetry is adjusted to about 0.5 McLett unit (about 10) 8 CFU/ml), and thenThe suspension was diluted 100-fold with MHB medium to a final concentration of about 10 6 CFU/ml。
3, experimental method:
a two-fold dilution of the microbillon recommended by the national Committee for standardization of clinical laboratories (ClinicaldLaboratyStandardsInstituteCLSI) was used.
The concentration of the experimental drug is set to be within the range of 128-0.008 mu g/ml by two-fold dilution, and the concentration is properly adjusted according to the actual situation.
A quantitative test drug is dissolved with DMSO and diluted by MHB culture medium to obtain a test drug solution with the concentration of 256 mug/ml. 200 μ l of each test drug solution was added to the first well of a 96-well plate. Mu.l MHB medium was added to each of the remaining wells, and the wells were sequentially diluted to 0.08. Mu.g/ml, 100. Mu.l of the solution was aspirated from the tail well and discarded. To obtain the pore plates with the drug concentrations of 128, 64, 32, 16, 8, 4, 2, 1, 0.5, 0.25, 0.125, 0.06, 0.03, 0.015 and 0.008 mu g/ml in sequence. Adding 100 μ l diluted corresponding bacterial solution into each hole, and mixing to obtain bacterial solution with final concentration of 5 × 10 5 CFU/ml. Separately, a test drug control group, a bacteria control group (100. Mu.l of a bacterial solution + 100. Mu.l of MHB medium), a vehicle bacteria control group (100. Mu.l of a bacterial solution + 100. Mu.l of DMSOMHB medium containing 1%), and a vehicle medium blank control group (200. Mu.l of DMSOMHB medium containing 1%) were prepared. After the plate is placed in an incubator at 35-37 ℃ for 20 hours, the A600 value of each hole is measured by a microplate reader, whether each hole has bacteria growth is judged, and the MIC value is determined.
4, experimental results:
the experiment selects 18-19 and 18-20 methicillin-resistant staphylococcus aureus (MRSA) as clinical separated pathogenic bacteria collected in 2018 Chengdu area. MRSA18-19 and MRSA18-20 are identified by a VITEK-60 automatic microorganism identifier and then identified by a conventional laboratory method. ATCC29213 is an outsourced instruction strain with certificate of identification.
The results of the activity measurements are shown in Table 1.
TABLE 1 in vitro antibacterial Activity of oxtriptolone-type ginsengenin derivatives
Figure BDA0002541728210000091
The antibacterial activity result shows that the inhibition rate of the 24 (R) -configuration ocotillol type ginsenoside OR of the oxcotion trombone type ginsenoside derivative on MRSA18-19, MRSA18-20 and ATCC29213 resistant methicillin staphylococcus aureus is more than 128 mu g/mL. Examples 1, 3, 5, 7 and 9 are derivatives having OR as the parent nucleus, and their MIC values are in the range of 2 to 64. Mu.g/mL. Among them, example 3 had the strongest inhibitory activity against MRSA18-20, MIC =2 μ g/mL, and was significantly stronger than the mother core OR.24 The inhibition rate of (S) -configuration ocotillol type ginsenoside OS on MRSA18-19, MRSA18-20 and ATCC29213 resistant methicillin staphylococcus aureus is 64 mu g/mL. In the examples obtained by using OS as a mother nucleus, the inhibition rates of example 2 to MRSA18-19, MRSA18-20 and ATCC29213 are all 1 mug/mL, and the inhibition effect to drug-resistant bacteria is obviously stronger than that of the mother nucleus.
Currently, drug-resistant staphylococcus aureus strains such as methicillin-resistant staphylococcus aureus (MRSA) are resistant to β -lactam antibiotics and resistant to many antibacterial drugs such as aminoglycosides, fluoroquinolones, macrolides, and the like. Moreover, the MRSA drug resistance develops rapidly, and compared with the quality control bacterial strain commonly used in a laboratory, the clinical separation pathogenic bacteria often have stronger drug resistance. However, the antibacterial activity result of the invention shows that the inhibition effect of the oxtriptolone type ginsenoside derivative obtained after structural modification and reformation on methicillin-resistant staphylococcus aureus is obviously enhanced.
Although the preferred embodiments of the present invention have been described in detail, the present invention is not limited to the details of the embodiments, and various equivalent modifications can be made within the technical spirit of the present invention, and the scope of the present invention is also within the scope of the present invention. It should be noted that the various features described in the above embodiments may be combined in any suitable manner without departing from the scope of the invention. The invention is not described in detail in order to avoid unnecessary repetition. In addition, any combination of the various embodiments of the present invention is also possible, and the same should be considered as the disclosure of the present invention as long as it does not depart from the spirit of the present invention.

Claims (1)

1. The application of the oxtripteryl-type ginsengenin derivative and the medically acceptable salt thereof in preparing methicillin-resistant staphylococcus aureus antibacterial drugs is characterized in that the oxtripteryl-type ginsengenin derivative is as follows:
Figure FDA0003839381650000011
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