CN104844617A - Brusatol derivatives and uses of brusatol derivatives in inflammations and immune function disorder diseases - Google Patents

Brusatol derivatives and uses of brusatol derivatives in inflammations and immune function disorder diseases Download PDF

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CN104844617A
CN104844617A CN201410054069.2A CN201410054069A CN104844617A CN 104844617 A CN104844617 A CN 104844617A CN 201410054069 A CN201410054069 A CN 201410054069A CN 104844617 A CN104844617 A CN 104844617A
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compound
arc
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brusatol
alkyl
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CN104844617B (en
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庾石山
侯琦
汤伟彬
袁绍鹏
徐嵩
吕海宁
屈晶
马双刚
白金叶
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Institute of Materia Medica of CAMS
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    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D493/00—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system
    • C07D493/02—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system in which the condensed system contains two hetero rings
    • C07D493/10—Spiro-condensed systems
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/4245—Oxadiazoles

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Abstract

The present invention discloses uses of a class of brusatol derivatives and uses of the brusatol derivatives in inflammations and immune function disorder diseases, particularly to a class of nitric oxide donor brusatol derivatives or medically acceptable salts thereof, a pharmaceutical composition containing the derivative, and applications of the derivatives or the salts thereof in preparation of anti-inflammatory and immunosuppressive drugs. In addition, the applications of the derivatives are provided in preparation of drugs for inflammations and/or immune disorder related diseases. The formula I is defined in the instruction.

Description

Brusatol derivative and the purposes in inflammation and immunologic function disorder disease thereof
Technical field
The present invention relates to biomedicine field, be specifically related to a class nitric oxide donors Brusatol derivative or its medically acceptable salt, the medicinal compositions containing these derivatives and they preparing the application in anti-inflammatory and immunosuppressive drug.
Background technology
Quassin is the natural product that a class has extensive pharmacologically active, and activity comprises antitumor, anti-malarial, anti-inflammatory, desinsection etc., but it generally has very strong toxicity.In quassin, crow courage booth Zeng Zuowei antitumor drug enters clinical study, but toxicity serious in body is one of reason of its failure.How to improve or keep active while, reducing toxicity is that quassinoids compound is used for clinical key.(I.H.Hall, K.H.Lee and etc, J.Pharm.Sci, 1983 according to the literature; 72:1282), quassinoids natural product Brusatol can stablize lysosome membrane, reduces the release of lytic enzyme, thus reduces the damage to surrounding tissue.In rat body on inflammatory model, Brusatol (brusatol) shows very strong anti-inflammatory activity, and activity is much better than indomethacin.But Brusatol has very strong toxicity, limit its application.
NO is endogenous compound, and it has physiological action widely.Research shows, at cell levels, continues NO (no matter the being endogenic or ectogenic) apoptosis capable of inhibiting cell of lower concentration in body, cell is had to the effect of protection and growth promoting effects.In Immunological diseases research, most scholar tends to the generation evolution that endogenous NO can participate in autoimmune disease, has immunoregulatory activity, white corpuscle and endothelial cell adhesion can be regulated, suppress thrombocyte, leukocytic gathering, suppressor T cell is bred, and improves the effect of NK cytoactive etc.At present, show according to existing research, the nitric oxide donors non-steroidal anti-inflammatory drugs (NO-NSAID) that NO donor (namely having molecule or the group of release NO ability) generates with NSAID coupling, in maintenance or while increasing NSAID curative effect (as anti-platelet activity increases), the untoward reaction (as gi tract and cardiovascular adverse effects) of NSAID obviously can be reduced.But NO also has dual function, in body, appropriate endogenous NO has the effect of inflammation-inhibiting on the one hand; On the other hand iNOS(iNOS in inflammatory process) activate, synthesizing excessive NO again can exacerbate inflammation further.
Therefore, by NO donor and Brusatol coupling, obtain nitric oxide donor type derivant, at the artificially controlled initial stage of inflammation in vivo, produce a small amount of NO, play similar endothelium-derived NO effect, carry out inflammation-inhibiting, thus suppress iNOS overexpression.The NO simultaneously continuing lower concentration also can improve the survival rate of cell under the toxic action of Brusatol, plays and falls hypotoxic effect.
At present, have no in prior art about to NO donator type Brusatol derivative or its medically acceptable salt, be used for the treatment of the report of the disease such as inflammation and immunologic function disorder.
Summary of the invention:
The present invention is that the shortcoming existed to overcome prior art proposes, and its technical problem solved is to provide the active high and Brusatol analog derivative that toxic side effect is little of a class.
For this reason, the invention discloses a class new compound, that is, the nitric oxide donator type Brusatol derivative as shown in general formula I:
Wherein, R 1, R 2, R 4independently be selected from H, the furazan ring replaced,
Wherein, the furazan ring of replacement is selected from or
Wherein, L 1be selected from R 6or-R 6-X 1-Y 1-, wherein, Y 1be connected with O in primer, R 6be selected from C 2-6alkyl, C 2-4oC 2-4alkyl, C 2-6thiazolinyl, C 2-6alkynyl, substituted or unsubstituted phenyl, the substituting group of wherein said phenyl is selected from C 1-6alkyl, X 1be selected from O or NH, Y 1be selected from succinyl or phthaloyl;
L 2be selected from R 7,-R 7-X 2-Y 2-, Y 2, wherein, Y 2be connected with O in primer, R 7be selected from substituted or unsubstituted phenyl, the substituting group of wherein said phenyl is selected from C 1-6alkyl, X 2be selected from O or NH, Y 2be selected from succinyl or phthaloyl;
R 3be selected from OH, OCH 3, replace furazan ring,
The furazan ring wherein replaced is selected from or
Wherein R 9be selected from C 2-6alkyl, C 2-4oC 2-4alkyl, C 2-6thiazolinyl, C 2-6alkynyl, substituted or unsubstituted phenyl, the substituting group of wherein said phenyl is selected from C 1-6alkyl;
L 3be selected from-O-R 10-O-or O, wherein R 10be selected from substituted or unsubstituted phenoxy group, wherein, CH on oxygen and furazan ring 2be connected, the substituting group of wherein said phenoxy group is selected from C 1-6alkyl;
According to the present invention, the compound shown in preferred general formula (I) comprises, but is not limited to the compound shown in general formula (Ia):
Wherein, R 3be selected from OH, OCH 3, L 1be selected from-R 6-X 1-Y 1-, wherein, Y 1be connected with O in primer, R 6be selected from C 2-6alkyl, C 2-4oC 2-4alkyl, C 2-6alkynyl ,-o-Ph-(CH 2) n-, wherein n=0,1,2,3,4 ,-m-Ph-(CH 2) n-, wherein n=0,1,2,3,4 ,-p-Ph-(CH 2) n-, wherein n=0,1,2,3,4; X 1be selected from O, Y 1be selected from succinyl or phthaloyl.
According to the present invention, the compound shown in preferred general formula (I) comprises, but is not limited to the compound shown in general formula (Ib):
Wherein, R 3be selected from OH, OCH 3, L 2be selected from R 7,-R 7-X 2-Y 2-, Y 2, wherein, Y 2be connected with O in primer, R 7be selected from-o-Ph-(CH 2) n-, wherein n=0,1,2,3,4 ,-m-Ph-(CH 2) n-, wherein n=0,1,2,3,4 ,-p-Ph-(CH 2) n-, wherein n=0,1,2,3,4; X 2be selected from O, Y 2be selected from succinyl or phthaloyl.
According to the present invention, the compound shown in preferred general formula (I) comprises, but is not limited to the compound shown in general formula (Ic):
Wherein, L 1be selected from-Y 1-X 1-R 6-, wherein, Y 1be connected with O in primer, R 6be selected from C 2-6alkyl, C 2-4oC 2-4alkyl, C 2-6alkynyl ,-(CH 2) n-o-Ph-, wherein n=0,1,2,3,4 ,-(CH 2) n-m-Ph-, wherein n=0,1,2,3,4 ,-(CH 2) n-p-Ph-, wherein n=0,1,2,3,4, wherein, phenyl connect CH 2all and X 1be connected; X 1be selected from O, Y 1be selected from succinyl or phthaloyl.
According to the present invention, the compound shown in preferred general formula (I) comprises, but is not limited to the compound shown in general formula (Id):
Wherein, L 2be selected from-Y 2-X 2-R 7-, wherein, Y 2be connected with O in primer, R 7be selected from-(CH 2) n-o-Ph-, wherein n=0,1,2,3,4 ,-(CH 2) n-m-Ph-, wherein n=0,1,2,3,4 ,-(CH 2) n-p-Ph-, wherein n=0,1,2,3,4, wherein, phenyl connect CH 2all and X 2be connected; X 2be selected from O, Y 2be selected from succinyl or phthaloyl.
According to the present invention, the compound shown in preferred general formula (I) comprises, but is not limited to the compound shown in general formula (Ie):
Wherein, R 9be selected from C 2-6alkyl, C 2-4oC 2-4alkyl, C 2-6alkynyl ,-(CH 2) n-o-Ph-, wherein n=0,1,2,3,4 ,-(CH 2) n-m-Ph-, wherein n=0,1,2,3,4 ,-(CH 2) n-p-Ph-, wherein n=0,1,2,3,4, wherein, phenyl connect CH 2all be connected with primer by O.
According to the present invention, the compound shown in preferred general formula (I) comprises, but is not limited to the compound shown in general formula (If):
Wherein, L 3be selected from-O-R 10-O-or O,
Wherein, R 10be selected from-(CH 2) n-o-Ph-, wherein n=0,1,2,3,4 ,-(CH 2) n-m-Ph-, wherein n=0,1,2,3,4 ,-(CH 2) n-p-Ph-, wherein n=0,1,2,3,4, wherein, phenyl connect CH 2all be connected with primer by O.
According to the present invention, the compound shown in preferred general formula (I) comprises, but is not limited to the compound shown in general formula (Ig):
Wherein, R 1be selected from H, l 1be selected from-R 6-X 1-Y 1-, wherein, Y 1all be connected with O in primer, R 6be selected from C 2-6alkyl, C 2-4oC 2-4alkyl, C 2-6alkynyl ,-o-Ph-(CH 2) n-, wherein n=0,1,2,3,4 ,-m-Ph-(CH 2) n-, wherein n=0,1,2,3,4 ,-p-Ph-(CH 2) n-, wherein n=0,1,2,3,4, wherein, phenyl connect CH 2all and X 1be connected; X 1be selected from O, Y 1be selected from succinyl or phthaloyl.
According to the present invention, the compound shown in preferred general formula (I) comprises, but is not limited to the compound shown in general formula (Ii):
Wherein, R 1be selected from H, l 2be selected from-R 7-X 2-Y 2-, Y 2, wherein, Y 2all be connected with O in primer, R 7be selected from-o-Ph-(CH 2) n-, wherein n=0,1,2,3,4 ,-m-Ph-(CH 2) n-, wherein n=0,1,2,3,4 ,-p-Ph-(CH 2) n-, wherein n=0,1,2,3,4, wherein, phenyl connect CH 2all and X 2be connected; X 2be selected from O; Y 2be selected from succinyl or phthaloyl.
Specifically, the nitric oxide donator type Brusatol derivative shown in general formula I preferably from but be not limited only to following compounds (compound numbers corresponds to the compound numbers in embodiment):
Compound R 6 X 1 Y 1
1 (CH 2) 3 O CO(CH 2) 2CO
2 (CH 2) 4 O CO(CH 2) 2CO
3 (CH 2) 2CH(CH 3) O CO(CH 2) 2CO
4 (CH 2) 5 O CO(CH 2) 2CO
5 (CH 2) 2O(CH 2) 2 O CO(CH 2) 2CO
6 CH 2C≡CCH 2 O CO(CH 2) 2CO
7 p-PhCH 2 O CO(CH 2) 2CO
8 m-PhCH 2, O CO(CH 2) 2CO
9 (CH 2) 4 O CO-o-PhCO
Compound R 7 X 2 Y 2
10 p-Ph-CH 2 O CO(CH 2) 2CO
11 m-Ph-CH 2 O CO(CH 2) 2CO
12 o-Ph-CH 2 O CO(CH 2) 2CO
13 - O CO(CH 2) 2CO
14 - O CO-o-PhCO
15 p-Ph-CH 2 - -
Compound R 6 X 1 Y 1
16 (CH 2) 3 O CO(CH 2) 2CO
17 (CH 2) 4 O CO(CH 2) 2CO
18 (CH 2) 2CH(CH 3) O CO(CH 2) 2CO
19 (CH 2) 5 O CO(CH 2) 2CO
20 (CH 2) 2O(CH 2) 2 O CO(CH 2) 2CO
21 CH 2C≡CCH 2 O CO(CH 2) 2CO
22 CH 2-p-Ph O CO(CH 2) 2CO
23 CH 2-m-Ph O CO(CH 2) 2CO
24 (CH 2) 4 O CO-o-PhCO
Compound R 7 X 2 Y 2
25 CH 2-p-Ph O CO(CH 2) 2CO
26 CH 2-m-Ph O CO(CH 2) 2CO
27 CH 2-o-Ph O CO(CH 2) 2CO
28 - O CO(CH 2) 2CO
29 - O CO-o-PhCO
Compound R 9
30 CH 2CH 2CH 2
31 CH 2CH 2CH 2CH 2
32 CH 2-p-Ph
33 CH 2-m-Ph
Compound L 3
34 OCH 2-p-PhO
35 OCH 2-m-PhO
36 OCH 2-o-PhO
37 O
The invention still further relates to a kind of containing the compound as described in situation as each in general formula I of medicine effective dose and the pharmaceutical composition of pharmaceutically acceptable carrier.
According to the present invention, the compounds of this invention can exist with the form of isomer, and usually described " the compounds of this invention " comprises the isomer of this compound.
According to embodiment of the present invention, described the compounds of this invention also comprises hydrate or the prodrug of acceptable salt, salt in its pharmacodynamics.
The invention still further relates to and contain as the compounds of this invention of active ingredient and the pharmaceutical composition of customary pharmaceutical excipients or assistant agent.Usual pharmaceutical composition of the present invention contains the compounds of this invention of 0.1 ~ 95% weight.In unit dosage form, the general content of the compounds of this invention is 0.1 ~ 100mg, and preferred unit dosage form contains 4 ~ 50mg.
The pharmaceutical composition of the compounds of this invention can be prepared according to method well known in the art.During for this object, if needed, the compounds of this invention and one or more solids or liquid pharmaceutical excipients and/or assistant agent can be combined, make the suitable administration form or dosage form that can be used as people's medicine or veterinary drug use.
The compounds of this invention or the pharmaceutical composition containing it can administrations in a unit, and route of administration can be enteron aisle or non-bowel, as oral, muscle, subcutaneous, nasal cavity, oral mucosa, skin, peritonaeum or rectum etc.The compounds of this invention or the route of administration containing its pharmaceutical composition can be drug administration by injection.Injection comprises intravenous injection, intramuscular injection, subcutaneous injection, intradermal injection and acupoint injection therapy etc.
Form of administration can be liquid dosage form, solid dosage.If liquid dosage form can be true solution class, colloidal type, particulate formulations, emulsion dosage form, mixed suspension form.Other formulations are tablet, capsule, dripping pill, aerosol, pill, pulvis, solution, suspensoid, emulsion, granule, suppository, lyophilized injectable powder etc. such as.
The compounds of this invention can be made ordinary preparation, also can be sustained release preparation, controlled release preparation, targeting preparation and various particulate delivery system.
Such as in order to unit dosage forms for administration is made tablet, various carrier well known in the art can be widely used.Example about carrier is, such as thinner and absorption agent, as starch, dextrin, calcium sulfate, lactose, N.F,USP MANNITOL, sucrose, sodium-chlor, glucose, urea, calcium carbonate, white bole, Microcrystalline Cellulose, pure aluminium silicate etc.; Wetting agent and tackiness agent, as water, glycerine, polyoxyethylene glycol, ethanol, propyl alcohol, starch slurry, dextrin, syrup, honey, glucose solution, mucialga of arabic gummy, gelatine size, Xylo-Mucine, lac, methylcellulose gum, potassiumphosphate, polyvinylpyrrolidone etc.; Disintegrating agent, such as dry starch, alginates, agar powder, laminaran, sodium bicarbonate and Citric Acid, calcium carbonate, polyoxyethylene sorbitol fatty acid ester, sodium laurylsulfonate, methylcellulose gum, ethyl cellulose etc.; Disintegration inhibitor, such as sucrose, Tristearoylglycerol, theobroma oil, hydrogenation wet goods; Absorption enhancer, such as quaternary ammonium salt, sodium lauryl sulphate etc.; Lubricant, such as talcum powder, silicon-dioxide, W-Gum, stearate, boric acid, whiteruss, polyoxyethylene glycol etc.Tablet can also be made coating tablet further, such as sugar coated tablet, thin membrane coated tablet, ECT, or double-layer tablets and multilayer tablet.
Such as in order to administration unit is made pill, various carrier well known in the art can be widely used.Example about carrier is, such as thinner and absorption agent, as glucose, lactose, starch, theobroma oil, hydrogenated vegetable oil, polyvinylpyrrolidone, single stearic acid glycerine lipoprotein, kaolin, talcum powder etc.; Tackiness agent, as gum arabic, tragacanth gum, gelatin, ethanol, honey, liquid sugar, rice paste or batter etc.; Disintegrating agent, as agar powder, dry starch, alginates, sodium laurylsulfonate, methylcellulose gum, ethyl cellulose etc.
Such as in order to administration unit is made capsule, effective constituent the compounds of this invention is mixed with above-mentioned various carriers, and the mixture obtained thus is placed in hard gelatine capsule or soft capsule.Also effective constituent the compounds of this invention can be made microcapsule, be suspended in aqueous medium and form suspensoid, also can load in hard capsule or make injection application.
Such as, the compounds of this invention is made injection preparation, as solution, suspensoid solution, emulsion, lyophilized injectable powder, this preparation can be moisture or non-water, can containing acceptable carrier, thinner, tackiness agent, lubricant, sanitas, tensio-active agent or dispersion agent in a kind of and/or multiple pharmacodynamics.As thinner can be selected from water, ethanol, polyoxyethylene glycol, 1,3-PD, the isooctadecanol of ethoxylation, polyoxygenated isooctadecanol, polyoxyethylene sorbitol fat, fat acid esters etc.In addition, in order to prepare isotonic injection liquid, appropriate sodium-chlor, glucose or glycerine can be added in injection preparation, in addition, conventional solubility promoter, buffer reagent, pH adjusting agent etc. can also be added.These auxiliary materials are that this area is commonly used.
In addition, as needs, also tinting material, sanitas, spices, correctives, sweeting agent or other material can be added in pharmaceutical preparation.
For reaching medication object, strengthen result for the treatment of, medicine of the present invention or pharmaceutical composition can with any known medication administrations.
The dosage of the compounds of this invention pharmaceutical composition depends on many factors, such as to prevent or the character of disease therapy and severity, the sex of patient or animal, age, body weight, personality and individual reaction, route of administration, administration number of times, therapeutic purpose, therefore therapeutic dose of the present invention can have large-scale change.In general, the using dosage of Chinese materia medica composition of the present invention well known to a person skilled in the art.Can according to actual drug quantity contained in preparation last in the compounds of this invention composition, in addition suitable adjustment, to reach the requirement of its treatment significant quantity, completes prevention of the present invention or therapeutic purpose.The Suitable dosage ranges of the every day of the compounds of this invention: the consumption of compound of the present invention is 0.001 ~ 100mg/Kg body weight, is preferably 0.1 ~ 60mg/Kg body weight, is more preferably 1 ~ 30mg/Kg body weight, most preferably is 2 ~ 15mg/Kg body weight.The compounds of this invention every day that adult patient is taken is 10 ~ 500mg, is preferably 20 ~ 100mg, once can take or point to take for 2 ~ 3 times; The dosage of children taking, according to every kg body weight 5 ~ 30mg, is preferably 10 ~ 20mg/kg body weight.Above-mentioned dosage can single dose form or be divided into several, and such as two, three or four dosage forms for administration, this is limited to the clinical experience of administration doctor and the dosage regimen for the treatment of means.Compound of the present invention or composition can be taken separately, or merge with other treatment medicine or symptomatic drugs and use.
Another object of the present invention is to provide a class nitric oxide donator type Brusatol derivative or its medically acceptable salt, the hydrate of salt or prodrug preparing in anti-inflammatory and immunosuppression or immunologic function disorder and relative disease medicine thereof and applies.
The relative diseases such as described inflammation and immunologic function disorder comprise rheumatoid arthritis, osteoarthritis, rheumatic arthritis, urarthritis, lupus erythematosus syndromes, bronchitis, bursitis, tenosynovitis, psoriasis, eczema, burn, dermatitis, inflammatory bowel, Ke Laoen is sick, gastritis, irritable bowel syndrome, ulcerative colitis, multiple sclerosis, Autoimmune Encephalomyelitis, colorectal carcinoma, arteritis nodosa, thyroiditis, rheumatic fever, gingivitis, periodontitis stomatocace, ephritis, the swelling occurred after damaging, myocardial ischemia, various infectious pneumonia, physics and chemistry pneumonia and allergy pneumonia, chronic obstructive pulmonary disease, asthma, proctalgia fugax and rectum split, liver and gall capsulitis, cholangitis sclerosing cholangitis, primary biliary cirrhosis and cholecystitis etc.This kind of disease universals on a cellular level show as: scavenger cell overactivity, produce excessive NO; Lymphocyte transformation is abnormal, causes immunologic function disorder.
Invention has been compound and the experiment of NO and compounds affect mouse spleen lymphocyte propagation, transformation experiment are generated to the scavenger cell suppressing to stimulate through LPS, illustrate that the Brusatol derivative of invention has from cell levels and suppress scavenger cell to generate excessive NO and suppress lymphocyte transformation to be the activity of T cell and B cell.Simultaneously by the compound of research invention on the impact of chmice acute otitis and allergic dermatitis, show that the Brusatol derivative invented still has good anti-inflammatory and immunosuppressive activity in testing in vivo further.In addition, also by asthma and chronic obstructive pneumonia two pathological models, prove that the compounds of this invention also has good therapeutic action to immunologic function disorder relative disease further.
All show from inside and outside the pharmacological results, Brusatol derivative of the present invention has good anti-inflammatory and immunosuppressive activity, and can be used for treating inflammation and/or immunologic function disorder relative disease, be the compound that the active high toxic side effect of a class is little, there is good Development volue.
Accompanying drawing explanation
Fig. 1 asthmatic model small mouse body weight gain situation
Fig. 2 is on the impact of asthmatic mouse pathological change
Fig. 3 is to the Elisa measurement result of mouse asthma SERUM IgE
The impact that Fig. 4 transforms mouse asthma spleen lymphocyte proliferation
Arneth's count result in Fig. 5 mouse asthma bronchoalveolar lavage fluid
Fig. 6 is to the Elisa measurement result of inflammatory factor in mouse asthma bronchoalveolar lavage fluid
Arneth's count result in Fig. 7 chronic obstructive pulmonary disease mouse bronchoalveolar lavage fluid
Fig. 8 mouse bronchoalveolar lavage fluid inflammatory factor secretion situation
Embodiment:
In order to illustrate the present invention further, provide a series of embodiment below, these embodiments are illustrative completely, and they are only used for specifically describing the present invention, not should be understood to limitation of the present invention.
The synthetic route of compound 1-9 in embodiment: (intermediate a 1-a 8middle R 6, X 1respectively with R in compound 1-8 6, X 1unanimously, b 1-b 9middle R 6, X 1, Y 1respectively with R in compound 1-9 6, X 1, Y 1unanimously)
The end product (compound numbers corresponds to the compound numbers in embodiment) of synthesis:
Compound numbers R 6 X 1 Y 1
1 (CH 2) 3 O CO(CH 2) 2CO
2 (CH 2) 4 O CO(CH 2) 2CO
3 (CH 2) 2CH(CH 3) O CO(CH 2) 2CO
4 (CH 2) 5 O CO(CH 2) 2CO
5 (CH 2) 2O(CH 2) 2 O CO(CH 2) 2CO
6 CH 2C≡CCH 2 O CO(CH 2) 2CO
7 p-PhCH 2 O CO(CH 2) 2CO
8 m-PhCH 2, O CO(CH 2) 2CO
9 (CH 2) 4 O CO-o-PhCO
Embodiment 1:
Step 1:3-(3-benzenesulfonyl-1,2,5-oxadiazole-2-oxide compound-4-) oxygen propyl alcohol (a 1) preparation
2.5g (32mmol) 1,3-PD is dissolved in 15mlTHF, adds the NaOH solution 6ml of 10% to it, drip containing 2.2g(6mmol under ice-water bath condition) the 30ml THF solution of 2-oxygen-3,4-dibenzenesulfonyl-1,2,5-oxadiazole (A).Drip and finish, change and react with stirring at room temperature.After 2h, reaction solution is poured in 200ml water, extract with ethyl acetate (3 × 200ml).Organic layer merges, saturated common salt water washing, then with anhydrous sodium sulfate drying.After drying, filter, filtrate concentrates.Silica gel column chromatography is separated, with ethyl acetate: sherwood oil (60 ~ 90 DEG C)=1:3 ~ 1:1 (V:V) gradient elution, obtains white powdery solids 1.0g, is compound a 1, yield 55.5%.
Step 2:4-[3-(2-oxygen-3-benzenesulfonyl-1,2,5-oxadiazole-4-oxygen)-propoxy-]-4-ketobutyric acid (b 1) preparation
Get a 1280mg(0.93mmol), Succinic anhydried 200mg(2mmol), DMAP is about 30mg(0.25mmol) dissolve with anhydrous methylene chloride 40ml, 40 DEG C of return stirring reaction 24h.After reaction terminates, respectively with hot water, saturated common salt water washing.Organic layer is with anhydrous sodium sulfate drying.After drying, filter, filtrate concentrates, and silica gel column chromatography is separated, and with methylene dichloride: methyl alcohol=50:0 ~ 50:1 gradient elution, obtains white powdery solids 330mg, is compound b 1, yield 88.4%.
Step 3:3-O-{4-[3-(2-oxygen-3-benzenesulfonyl-1,2,5-oxadiazole-4-oxygen)-propoxy-]-4-oxobutanoyl } preparation of-Brusatol (1)
By EDCI86mg(0.44mmol), intermediate b 1140mg(0.35mmol) be placed in reaction flask, in ice-water bath, add anhydrous methylene chloride and dissolve, stir after 10 minutes, add Brusatol 90mg(0.17mmol) and DMAP23mg(0.19mmol), continue stirring reaction 3h under ice bath, then change with room temperature reaction 48h.Reaction solution respectively with rare HCl, rare NaHCO 3solution, saturated common salt water washing, organic layer is with anhydrous sodium sulfate drying.After drying, filter, filtrate concentrates, and silica gel column chromatography is separated, and with methylene dichloride: methyl alcohol=60:1 wash-out, obtains white powdery solids and be about 50mg, is compound 1, yield 32.0%.The physical and chemical parameter of compound 1 is as follows:
1H NMR(600MHz,CD 3COCD 3)δ8.07(2H,br d,J=7.8Hz,ArH),7.87(1H,t,J=7.8Hz，ArH),7.75(2H,t,J=7.8Hz,ArH),6.12(1H,br s,H-15),5.63(1H,s,H-2’),4.98(1H,s,H-7),4.73(1H,d,J=7.8Hz,H a-20),4.55(2H,t,J=6.0Hz，OCH 2),4.29(2H,t,J=6.0Hz，OCH 2),4.26(2H,overlap,H-11,H-12),4.13(2H,t,J=6.5Hz，OCH 2),3.78(1H,d,J=7.8Hz,H b-20),3.70(3H,s,CH 3O-21),3.33(1H,br s,H-14),3.24(1H,d,J=12.6Hz,H-5),2.85(2H,m,COCH 2),2.81(1H,d,J=16.0Hz,H β-1),2.79(1H,d,J=16.0Hz,H α-1),2.71(2H,t,J=6.0Hz,COCH 2),2.39(1H,br s,H-9),2.29(1H,dt,J=15.0,2.4Hz,H α-6),2.22(2H,m,CH 2),2.14(3H,H-5’),1.98(1H,ddd,J=15.0,12.6,2.4Hz,H β-6),1.91(3H,s,H-4’),1.82(3H,s,H-18),1.49(3H,s,H-19);
13C NMR(150MHz,CD 3COCD 3)δ189.9(C-2),172.5(C=O),171.7(C-21),170.8(C=O),167.6(C-16),164.9(C-1’),160.0(C-3’),159.0(C=N),146.8(C-3),142.7(C-4),139.0(ArC),136.7(ArC),130.7(ArC×2),129.4(ArC×2),115.8(C-2’),111.5(C=N),83.2(C-7),82.3(C-13),76.4(C-12),74.0(C-20),72.7(C-11),69.0(OCH 2),67.6(C-15),61.3(OCH 2),52.7(C-21),51.2(C-14),50.4(C-1),46.1(C-8),43.2(C-5),41.6(C-9),41.5(C-10),29.6(CO CH 2),29.4(CO CH 2),29.1(C-6),28.6(CH 2),27.2(C-4’),20.2(C-5’),15.8(C-19),14.5(C-18)；HRESIMS m/z903.2476[M+H] +(calcd for C 41H 47N 2O 19S,903.2488).
Embodiment 2:
3-O-{4-[4-(2-oxygen-3-benzenesulfonyl-1,2,5-oxadiazole-4-oxygen)-butoxy]-4-oxobutanoyl }-Brusatol (2)
Step 1: with reference to step 1 in embodiment 1, replaces 1,3-PD and 2-oxygen-3,4-dibenzenesulfonyl-1,2,5-oxadiazole to react with BDO, obtains 4-(3-benzenesulfonyl-1,2,5-oxadiazole-2-oxide compound-4-) oxygen butanols (a 2).
Step 2: with reference to step 2 in embodiment 1, with a 2replace a 1, be obtained by reacting intermediate 4-[4-(2-oxygen-3-benzenesulfonyl-1,2,5-oxadiazole-4-oxygen)-butoxy]-4-ketobutyric acid (b with Succinic anhydried 2).
Step 3: with reference to step 3 in embodiment 1, with b 2replace b 1, under the effect of excessive EDCI and DMAP, reacting obtained white powdery solids with Brusatol, is compound 2, yield 65.5%.
The physical and chemical parameter of compound 2 is as follows:
1H NMR(400MHz,CDCl 3)δ8.05(2H,br d,J=7.6Hz，ArH),7.76(1H,t,J=7.6Hz,ArH),7.62(2H,t,J=7.6Hz,ArH),6.24(1H,br s,H-15),5.62(1H,s,H-2’),4.79(1H,s,H-7),4.72(1H,d,J=7.6Hz,H a-20),4.44(2H,t,J=6.0Hz，OCH 2),4.24(1H,d,J=2.8Hz,H-11),4.20(2H,t,J=6.0Hz，OCH 2),4.19(1H,s,H-12),3.79(1H,d,J=7.6Hz,H b-20),3.78(3H,s,CH 3O-21),3.14(1H,br s,H-14),3.04(1H,d,J=12.4Hz,H-5),2.95(1H,d,J=16.0Hz,H β-1),2.88(2H,t,J=6.8Hz,COCH 2),2.72(2H,m,COCH 2),2.39(1H,d,J=16.0Hz,H α-1),2.36(1H,d,J=14.0Hz,H α-6),2.19(3H,s,H-5’),2.09(1H,br s,H-9),1.97(2H,m,CH 2),1.93(3H,s,H-4’),1.84(2H,m,CH 2),1.84(1H,overlap,H β-6),1.81(3H,s,H-18),1.47(3H,s,H-19);
13C NMR(100MHz,CDCl 3)δ188.9(C-2),172.0(C=O),171.9(C-21),170.2(C=O),166.9(C-16),164.4(C-1’),161.0(C-3’),158.9(C=N),145.6(C-3),142.1(C-4),138.0(ArC),135.6(ArC),129.7(ArC×2),128.6(ArC×2),114.0(C-2’),110.5(C=N),82.1(C-7),81.2(C-13),75.8(C-12),73.9(C-20),71.0(OCH 2),70.8(C-11),65.7(C-15),63.9(OCH 2),53.0(OCH 3),51.6(C-14),50.1(C-1),45.3(C-8),42.8(C-5),41.7(C-9),40.7(C-10),29.0(C-6),28.8(CO CH 2),28.5(CO CH 2),27.7(C-4’),25.1(CH 2),25.0(CH 2),20.6(C-5’),15.4(C-19),14.5(C-18)；
HRESIMS m/z939.2450[M+Na] +(calcd for C 42H 48N 2N aO 19S,939.2464).
Embodiment 3:
(±)-3-O-{4-[(2-oxygen-3-benzenesulfonyl-1,2,5-oxadiazole-4-oxygen)-3-butoxy]-4-oxobutanoyl }-Brusatol (3)
Step 1: with reference to step 1 in embodiment 1, replaces 1,3-PD and 2-oxygen-3,4-dibenzenesulfonyl-1,2,5-oxadiazole to react with 1,3 butylene glycol, obtains (3-benzenesulfonyl-1,2,5-oxadiazole-2-oxide compound-4-) oxygen-3-butanols (a 3).
Step 2: with reference to step 2 in embodiment 1, with a 3replace a 1, be obtained by reacting intermediate 4-[(2-oxygen-3-benzenesulfonyl-1,2,5-oxadiazole-4-oxygen)-3-butoxy]-4-ketobutyric acid (b with Succinic anhydried 3).
Step 3: with reference to step 3 in embodiment 1, with b 3replace b 1, under the effect of excessive EDCI and DMAP, reacting obtained white powdery solids with Brusatol, is compound 3, yield 50.0%.
The physical and chemical parameter of compound 3 is as follows:
1h NMR (600MHz, CD 3cOCD 3) (be raceme, part hydrogen display m peak, therefore peak shape is not indicated) δ 8.06 (2H, ArH), 7.87 (1H, ArH), 7.75 (2H, ArH), 6.11 (1H, H-15), 5.63 (1H, H-2 '), 5.17 (1H, OCH), 4.97 (1H, H-7), 4.73 (1H, H a-20), 4.51 (2H, OCH 2), 4.26 (2H, H-11, H-12), 3.78 (1H, H b-20), 3.69 (3H, CH 3o-21), 3.32 (1H, H-14), 3.23 (1H, H-5), 2.85 (2H, COCH 2), 2.78 (1H, H β-1), 2.72 (1H, H α-1), 2.69 (2H, COCH 2), 2.39 (1H, H-9), 2.28 (1H, H α-6), 2.15 (2H, CH 2), 2.14 (3H, H-5 '), 1.96 (1H, H β-6), 1.91 (3H, H-4 '), 1.79 (3H, H-18), 1.48 (3H, H-19), 1.32 (3H, CH 3);
13C NMR(150MHz,CD 3COCD 3)δ189.9(C-2),172.2(C=O),171.7(C-21),170.8(C=O),167.6(C-16),164.9(C-1’),160.0(C-3’),159.0(C=N),146.8(C-3),142.7(C-4),139.0(ArC),136.7ArC),130.7(ArC×2),129.4(ArC×2),115.8(C-2’),111.5(C=N),83.2(C-7),82.3(C-13),76.4(C-12),74.0(C-20),72.7(C-11),71.7(OCH),68.8(OCH 2),68.4(OCH 2),67.7(C-15),52.7(C-21),51.3(C-14),50.4(C-1),46.1(C-8),43.2(C-5),41.6(C-9),41.5(C-10),35.4(CH 3),29.9(CO CH 2),29.1(C-6),27.2(C-4’),20.2(C-5’),15.7(C-19),14.5(C-18)；
HRESIMS m/z939.2475[M+Na] +(calcd for C 42H 48N 2N aO 19S,939.2464).
Embodiment 4:
3-O-{4-[5-(2-oxygen-3-benzenesulfonyl-1,2,5-oxadiazole-4-oxygen)-pentyloxy]-4-oxobutanoyl }-Brusatol (4)
Step 1: with reference to step 1 in embodiment 1, replaced by 1,5-PD 1,3-PD and 2-oxygen-3,4-dibenzenesulfonyl-1,2,5-oxadiazole to react, obtain 5-(3-benzenesulfonyl-1,2,5-oxadiazole-2-oxide compound-4-) oxygen-amylalcohol (a 4).
Step 2: with reference to step 2 in embodiment 1, with a 4replace a 1, be obtained by reacting intermediate 4-[5-(2-oxygen-3-benzenesulfonyl-1,2,5-oxadiazole-4-oxygen)-pentyloxy]-4-ketobutyric acid (b with Succinic anhydried 4).
Step 3: with reference to step 3 in embodiment 1, with b 4replace b 1, then under the effect of excessive EDCI and DMAP, reacting obtained white powdery solids with Brusatol, is compound 4, yield 56.5%.
The physical and chemical parameter of compound 4 is as follows:
1H NMR(500MHz,CD 3COCD 3)δ8.06(2H,dd,J=8.0,1.0Hz,ArH),7.88(1H,t,J=8.0Hz,ArH),7.75(2H,dd,J=8.0,8.0Hz,ArH),6.10(1H,br s,H-15),5.63(1H,s,H-2’),4.98(1H,s,H-7),4.73(1H,d,J=7.5Hz,H a-20),4.46(2H,t,J=6.5Hz,OCH 2),4.27(1H,overlap,H-11),4.26(1H,d,J=4.5Hz,H-12),4.13(2H,t,J=6.5Hz,OCH 2),3.78(1H,d,J=7.5Hz,H b-20),3.69(3H,s,CH 3O-21),3.32(1H,br s,H-14),3.25(1H,d,J=12.5Hz,H-5),2.85(2H,m,COCH 2),2.81(1H,d,J=16.0Hz,H β-1),2.72(1H,d,J=16.0Hz,H α-1),2.69(2H,t,J=6.5Hz,COCH 2),2.40(1H,br s,H-9),2.29(1H,dt,J=14.0,2.5Hz,H α-6),2.13(3H,s,H-5’),2.01(1H,ddd,J=14.0,12.5,2.5Hz,H β-6),1.91(3H,s,H-4’),1.89(2H,m,CH 2),1.83(3H,s,H-18),1.73(2H,m,CH 2),1.55(2H,m,CH 2),1.50(3H,s,H-19);
13C NMR(125MHz,CD 3COCD 3)δ189.9(C-2),172.5(C=O),171.7(C-21),170.8(C=O),167.6(C-16),165.0(C-1’),160.1(C-3’),159.0(C=N),146.8(C-3),142.7(C-4),139.1(ArC),136.7(ArC),130.7(ArC×2),129.3(ArC×2),115.8(C-2’),111.4(C=N),83.2(C-7),82.3(C-13),76.4(C-12),74.0(C-20),72.6(C-11),72.1(OCH 2),67.5(C-15),64.8(OCH 2),52.7(C-21),51.1(C-14),50.4(C-1),46.1(C-8),43.2(C-5),41.6(C-9),41.5(C-10),29.6(CO CH 2),29.4(CO CH 2),29.1(C-6),28.8(CH 2),28.7(CH 2),27.2(C-4’),22.8(CH 2),20.2(C-5’),15.7(C-19),14.5(C-18)；
HRESIMS m/z953.2610[M+Na] +(calcd for C 43H 50N 2N aO 19S,953.2621).
Embodiment 5:
3-O-{4-{2-{2-[(2-oxygen 3-benzenesulfonyl-1,2,5-oxadiazole-4-oxygen) oxyethyl group] }-oxyethyl group }-4-oxobutanoyl }-Brusatol (5)
Step 1: with reference to step 1 in embodiment 1, replaces 1,3-PD and 2-oxygen-3 with glycol ether; 4-dibenzenesulfonyl-1,2,5-oxadiazole reacts; obtain 2-{2-[(3-benzenesulfonyl-1,2,5-oxadiazole-2-oxide compound-4-) oxygen oxyethyl group] }-ethanol (a 5).
Step 2: with reference to step 2 in embodiment 1, with a 5replace a 1, be obtained by reacting intermediate 4-{2-{2-[(2-oxygen 3-benzenesulfonyl-1,2,5-oxadiazole-4-oxygen) oxyethyl group] with Succinic anhydried } and-oxyethyl group }-4-ketobutyric acid (b 5).
Step 3: with reference to step 3 in embodiment 1, with b 5replace b 1, under the effect of excessive EDCI and DMAP, reacting obtained white powdery solids with Brusatol, is compound 5, yield 47.2%.
The physical and chemical parameter of compound 5 is as follows:
1H NMR(500MHz,CDCl 3)δ8.06(2H,dd,J=7.5，1.0Hz，ArH),7.75(1H,t,J=7.5Hz,ArH),7.62(2H,t,J=7.5Hz,ArH),6.24(1H,br s,H-15),5.62(1H,s,H-2’),4.79(1H,s,H-7),4.72(1H,d,J=8.0Hz,H a-20),4.56(2H,m，OCH 2),4.30(2H,m，OCH 2),4.25(1H,s,H-11),4.19(1H,s,H-12),3.91(2H,m，OCH 2),3.79(1H,overlap,H b-20),3.79(2H,m，OCH 2),3.78(3H,s,CH 3O-21),3.15(1H,br s,H-14),3.04(1H,d,J=12.5Hz,H-5),2.95(1H,d,J=16.0Hz,H β-1),2.88(2H,t,J=6.8Hz,COCH 2),2.74(2H,m,COCH 2),2.39(1H,d,J=16.0Hz,H α-1),2.38(1H,dt,J=14.0Hz,2.5Hz,H α-6),2.19(3H,d,J=1.0Hz,H-5’),2.09(1H,br s,H-9),1.93(3H,d,J=1.0Hz,H-4’),1.82(1H,ddd,J=14.0,12.5,2.5Hz,H β-6),1.80(3H,d,J=1.0Hz，H-18),1.47(3H,s,H-19);
13C NMR(125MHz,CDCl 3)δ188.9(C-2),172.0(C=O),171.9(C-21),170.2(C=O),166.9(C-16),164.4(C-1’),161.0(C-3’),158.9(C=N),145.7(C-3),142.1(C-4),138.0(ArC),135.6(ArC),129.7(ArC×2),128.5(ArC×2),114.0(C-2’),110.5(C=N),82.1(C-7),81.2(C-13),75.8(C-12),73.9(C-20),70.8(C-11),70.5(OCH 2),69.3(OCH 2),68.4(OCH 2),65.7(C-15),63.8(OCH 2),53.0(OCH 3),51.6(C-14),50.1(C-1),45.3(C-8),42.8(C-5),41.7(C-9),40.7(C-10),28.9(C-6),28.8(CO CH 2),28.4(CO CH 2),27.6(C-4’),20.6(C-5’),15.4(C-19),14.4(C-18)；
HRESIMS m/z955.2418[M+Na] +(calcd for C 42H 48N 2N aO 20S,955.2413).
Embodiment 6:
3-O-{4-[4-(2-oxygen-3-benzenesulfonyl-1,2,5-oxadiazole-4-oxygen)-2-alkynes-butoxy]-4-oxobutanoyl }-Brusatol (6)
Step 1: with reference to step 1 in embodiment 1, replaces 1,3-PD and 2-oxygen-3 with Isosorbide-5-Nitrae butynediol; 4-dibenzenesulfonyl-1,2,5-oxadiazole reacts; obtain 4-(3-benzenesulfonyl-1,2,5-oxadiazole-2-oxide compound-4-) oxygen-2-alkynes-butanols (a 6).
Step 2: with reference to step 2 in embodiment 1, with a 6replace a 1, be obtained by reacting intermediate 4-[4-(2-oxygen-3-benzenesulfonyl-1,2,5-oxadiazole-4-oxygen)-2-alkynes-butoxy]-4-ketobutyric acid (b with Succinic anhydried 6).
Step 3: with reference to step 3 in embodiment 1, with b 6replace b 1, under the effect of excessive EDCI and DMAP, reacting obtained white powdery solids with Brusatol, is compound 6, yield 45.0%.
The physical and chemical parameter of compound 6 is as follows:
1H NMR(500MHz,CD 3COCD 3)δ8.07(2H,dd,J=8.0，1.0Hz,ArH),7.89(1H,t,J=7.5Hz，ArH),7.76(2H,dd,J=8.0，7.5Hz,ArH),6.12(1H,br s,H-15),5.63(1H,s,H-2’),5.23(2H,s,OCH 2),4.98(1H,s,H-7),4.83(2H,s,OCH 2),4.73(1H,d,J=7.5Hz,H a-20),4.26(2H,overlap,H-11,H-12),3.78(1H,d,J=7.5Hz,H b-20),3.69(3H,s,CH 3O-21),3.32(1H,br s,H-14),3.26(1H,d,J=12.5Hz,H-5),2.86(2H,m,COCH 2),2.82(1H,d,J=16.5Hz,H β-1),2.74(2H,t,J=6.5Hz,COCH 2),2.73(1H,overlap,H α-1),2.41(1H,br s,H-9),2.30(1H,dt,J=15.0,2.5Hz,H α-6),2.14(3H,s,H-5’),1.99(1H,ddd,J=14.0,12.5,2.5Hz,H β-6),1.91(3H,s,H-4’),1.84(3H,d,J=1.0Hz,H-18),1.50(3H,s,H-19);
13C NMR(125MHz,CD 3COCD 3)δ189.9(C-2),171.9(C=O),171.7(C-21),170.6(C=O),167.6(C-16),164.9(C-1’),159.2(C-3’),159.0(C=N),146.9(C-3),142.7(C-4),138.9(ArC),136.8(ArC),130.7(ArC×2),129.4(ArC×2),115.8(C-2’),111.6(C=N),84.6(C≡C,1C),83.2(C-7),82.3(C-13),79.8(C≡C,1C),76.4(C-12),74.0(C-20),72.7(C-11),67.6(C-15),59.5(CH 2),52.7(C-21),52.6(CH 2)50.9(C-14),50.4(C-1),46.1(C-8),43.3(C-5),41.7(C-9),41.5(C-10),29.5(CO CH 2),29.4(CO CH 2),29.0(C-6),27.2(C-4’),20.2(C-5’),15.8(C-19),14.5(C-18)；
HRESIMS m/z935.2165[M+Na] +(calcd for C 42H 44N 2N aO 19S,935.2151).
Embodiment 7:
3-O-{4-[4-(2-oxygen-3-benzenesulfonyl-1,2,5-oxadiazole-4 oxygen)-benzyloxy]-4-oxobutanoyl }-Brusatol (7)
Step 1: with reference to step 1 in embodiment 1, replaces 1,3-PD and 2-oxygen-3,4-dibenzenesulfonyl-1,2,5-oxadiazole to react with p-Hydroxybenzylalcohol, obtains 4-(3-benzenesulfonyl-1,2,5 oxadiazole-2-oxide compound-4-) methoxybenzyl (a 7).
Step 2: with reference to step 2 in embodiment 1, with a 7replace a 1, be obtained by reacting intermediate 4-[4-(2-oxygen-3-benzenesulfonyl-1,2,5-oxadiazole-4 oxygen)-benzyloxy]-4-ketobutyric acid (b with Succinic anhydried 7).
Step 3: with reference to step 3 in embodiment 1, with b 7replace b 1, under the effect of excessive EDCI and DMAP, reacting obtained white powdery solids with Brusatol, is compound 7, yield 47.4%.
The physical and chemical parameter of compound 7 is as follows:
1H NMR(500MHz,CD 3COCD 3)δ8.08(2H,dd,J=7.5,1.0Hz,ArH),7.90(1H,t,J=7.5Hz，ArH),7.75(2H,dd,J=7.5,7.5Hz,ArH),7.53(2H,d,J=8.5Hz,ArH),7.42(2H,d,J=8.5Hz,ArH),6.10(1H,br s,H-15),5.63(1H,s,H-2’),5.19(2H,t,J=13.0Hz,OCH 2),4.97(1H,s,H-7),4.74(1H,d,J=7.5Hz,H a-20),4.26(2H,overlap,H-11,H-12),3.78(1H,d,J=7.5Hz,H b-20),3.69(3H,s,CH 3O-21),3.33(1H,br s,H-14),3.25(1H,d,J=13.0Hz,H-5),2.86(2H,m,COCH 2),2.81(1H,overlap,H β-1),2.76(2H,t,J=6.5Hz,COCH 2),2.73(1H,d,J=16.5Hz,H α-1),2.41(1H,br s,H-9),2.29(1H,dt,J=14.5,2.5Hz,H α-6),2.14(3H,s,H-5’),2.01(1H,ddd,J=2.5,13.0,14.0Hz,H β-6),1.91(3H,s,H-4’),1.78(3H,d,J=1.0Hz,H-18),1.50(3H,s,H-19);
13C NMR(125MHz,CD 3COCD 3)δ189.9(C-2),172.4(C=O),171.7(C-21),170.7(C=O),167.6(C-16),165.0(C-1’),159.6(C-3’),159.0(C=N),153.7(ArC),146.9(C-3),142.7(C-4),138.8(ArC),136.8(ArC),135.9(ArC),130.7(ArC×2),130.7(ArC×2),129.5(ArC×2),120.6(ArC×2),115.9(C-2’),112.1(C=N),83.2(C-7),82.3(C-13),76.5(C-12),74.0(C-20),72.7(C-11),67.6(C-15),65.9(OCH 2),52.7(C-21),51.1(C-14),50.5(C-1),46.1(C-8),43.3(C-5),41.7(C-9),41.6(C-10),30.2(CO CH 2),30.0(CO CH 2),29.1(C-6),27.2(C-4’),20.2(C-5’),15.8(C-19),14.5(C-18)；
HRESIMS m/z973.2310[M+Na] +(calcd for C 45H 46N 2N aO 19S,973.2308).
Embodiment 8:
3-O-{4-[3-(2-oxygen-3-benzenesulfonyl-1,2,5-oxadiazole-4 oxygen)-benzyloxy]-4-oxobutanoyl }-Brusatol (8)
Step 1: with reference to step 1 in embodiment 1, replaces 1,3-PD and 2-oxygen-3,4-dibenzenesulfonyl-1,2,5-oxadiazole to react with a salicylic alcohol, obtains 3-(3-benzenesulfonyl-1,2,5 oxadiazole-2-oxide compound-4-) methoxybenzyl (a 8).
Step 2: with reference to step 2 in embodiment 1, with a 8replace a 1, be obtained by reacting intermediate 4-[3-(2-oxygen-3-benzenesulfonyl-1,2,5-oxadiazole-4 oxygen)-benzyloxy]-4-ketobutyric acid (b with Succinic anhydried 8).
Step 3: with reference to step 3 in embodiment 1, with b 8replace b 1, under the effect of excessive EDCI and DMAP, reacting obtained white powdery solids with Brusatol, is compound 8, yield 46.2%.
The physical and chemical parameter of compound 8 is as follows:
1H NMR(500MHz,CD 3COCD 3)δ8.07(2H,dd,J=7.5,1.5Hz,ArH),7.90(1H,t,J=7.5Hz，ArH),7.75(2H,dd,J=7.5,7.5Hz,ArH),7.50(1H,t,J=7.5Hz,ArH),7.45(1H,s,ArH),7.40(2H,m,ArH),6.10(1H,br s,H-15),5.63(1H,s,H-2’),5.19(2H,s,OCH 2),4.97(1H,s,H-7),4.74(1H,d,J=7.5Hz,H a-20),4.27(2H,overlap,H-11,H-12),3.78(1H,d,J=7.5Hz,H b-20),3.70(3H,s,CH 3O-21),3.33(1H,br s,H-14),3.24(1H,d,J=13.0Hz,H-5),2.87(2H,m,COCH 2),2.80(1H,overlap,H β-1),2.77(2H,t,J=6.5Hz,COCH 2),2.70(1H,d,J=16.0Hz,H α-1),2.40(1H,br s,H-9),2.29(1H,dt,J=15.0,2.5Hz,H α-6),2.14(3H,s,H-5’),1.98(1H,ddd,J=15.0,13.0,2.5Hz,H β-6),1.91(3H,s,H-4’),1.78(3H,d,J=1.0Hz,H-18),1.49(3H,s,H-19);
13C NMR(125MHz,CD 3COCD 3)δ189.9(C-2),172.3(C=O),171.7(C-21),170.7(C=O),167.6(C-16),165.0(C-1’),159.6(C-3’),159.0(C=N),154.1(ArC),146.9(C-3),142.7(C-4),140.0(ArC),138.8(ArC),136.8(ArC),131.0(ArC),130.7(ArC×2),129.5(ArC×2),126.7(ArC),120.1(ArC),119.9(ArC),115.8(C-2’),112.1(C=N),83.2(C-7),82.3(C-13),76.5(C-12),74.0(C-20),72.7(C-11),67.5(C-15),65.8(OCH 2),52.7(C-21),50.9(C-14),50.4(C-1),46.1(C-8),43.2(C-5),41.7(C-9),41.5(C-10),30.2(CO CH 2),30.1(CO CH 2),29.1(C-6),27.2(C-4’),20.2(C-5’),15.8(C-19),14.5(C-18)；
HRESIMS m/z951.2490[M+H] +(calcd for C 45H 47N 2O 19S,951.2488).
Embodiment 9:
3-O-{2-[4-(2-oxygen-3-benzenesulfonyl-1,2,5-oxadiazole-4-oxygen)-butoxy carbonyl]-benzoyl }-Brusatol (9)
Step 1: with reference to step 1 in embodiment 1, replaces 1,3-PD and 2-oxygen-3,4-dibenzenesulfonyl-1,2,5-oxadiazole to react with BDO, obtains 4-(3-benzenesulfonyl-1,2,5-oxadiazole-2-oxide compound-4-) oxygen butanols (a 2).
Step 2: with reference to step 2 in embodiment 1, with a 2replace a 1, replace Succinic anhydried with Tetra hydro Phthalic anhydride, be obtained by reacting intermediate 2-[4-(2-oxygen-3-benzenesulfonyl-1,2,5-oxadiazole-4-oxygen)-butoxy carbonyl]-phenylformic acid (b 9).
Step 3: with reference to step 3 in embodiment 1, with b 9replace b 1, under the effect of excessive EDCI and DMAP, reacting obtained white powdery solids with Brusatol, is compound 9, yield 25.0%.
The physical and chemical parameter of compound 9 is as follows:
1H NMR(500MHz,CD 3COCD 3)δ8.07(2H,br d,J=7.5Hz,ArH),8.06(1H,overla p,ArH),7.84(1H,t,J=7.5Hz，ArH),7.81(1H,m，ArH),7.71(4H,m,ArH),6.12(1H,br s,H-15),5.64(1H,s,H-2’),4.99(1H,s,H-7),4.75(1H,d,J=7.5Hz,H a-20),4.51(2H,t,J=6.0Hz，OCH 2),4.39(2H,m，OCH 2),4.27(2H,overlap,H-11,H-12),3.79(1H,d,J=7.5Hz,H b-20),3.70(3H,s,CH 3O-21),3.33(1H,overlap,H-14),3.33(1H,d,J=12.5Hz,H-5),2.88(1H,d,J=16.0Hz,H β-1),2.81(1H,d,J=16.0Hz,H α-1),2.44(1H,br s,H-9),2.34(1H,dt,J=14.5,2.5Hz,H α-6),2.14(3H,s,H-5’),2.04(1H,overlap,H β-6),2.00(2H,m,CH 2),1.98(3H,s,H-4’),1.94(2H,m,CH 2),1.91(3H,s,H-18),1.57(3H,s,H-19);
13C NMR(125MHz,CD 3COCD 3)δ189.9(C-2),171.7(C-21),167.8(C=O),167.7(C-16),165.5(C=O),165.0(C-1’),160.1(C-3’),159.0(C=N),147.8(C-3),142.7(C-4),139.1(ArC),136.7(ArC),133.6(ArC),132.7(ArC),132.1(ArC),131.8(ArC),130.7(ArC×2),130.5(ArC),129.7(ArC),129.3(ArC×2),115.8(C-2’),111.5(C=N),83.2(C-7),82.3(C-13),76.4(C-12),74.0(C-20),72.7(C-11),71.9(OCH 2),67.5(C-15),65.6(OCH 2),52.7(C-21),50.9(C-14),50.5(C-1),46.1(C-8),43.4(C-5),41.7(C-9),41.5(C-10),29.1(C-6),27.2(C-4’),25.9(CH 2),25.6(CH 2),20.2(C-5’),15.9(C-19),14.8(C-18)；
HRESIMS m/z987.2477[M+Na] +(calcd for C 46H 48N 2N aO 19S,987.2464).
The synthetic route of compound 10-14 in embodiment: (intermediate c 1-c 3middle R 7, X 2respectively with R in compound 10-12 7, X 2unanimously, d 1-d 5middle R 7, X 2, Y 2respectively with R in compound 10-14 7, X 2, Y 2unanimously)
The end product (compound numbers corresponds to the compound numbers in embodiment) of synthesis:
Compound numbers R 7 X 2 Y 2
10 p-Ph-CH 2 O CO(CH 2) 2CO
11 m-Ph-CH 2 O CO(CH 2) 2CO
12 o-Ph-CH 2 O CO(CH 2) 2CO
13 - O CO(CH 2) 2CO
14 - O CO-o-PhCO
Embodiment 10:
The preparation of step 1:2-oxygen-3-methylol-4-phenyl-1,2,5-oxadiazole (B)
Added in 3mL Glacial acetic acid by 2.00g (15mmol) styryl carbinol, stirring at room temperature is dissolved to it, drips containing 3.00g (72mmol) NaNO in condition of ice bath downhill reaction liquid 2saturated aqueous solution, dropwise rear stirring at room temperature reaction 5h.After reaction terminates, add 20ml water in reaction solution, with ethyl acetate (3 × 50ml) extraction, merge organic layer, then use 5%NaOH solution (3 × 15mL) successively, saturated nacl aqueous solution (3 × 20ml) washs, anhydrous sodium sulfate drying.After drying, filter, filtrate concentrates, and silica gel column chromatography is separated, and with dichloromethane eluent, obtaining orange/yellow solid 1.2g, is compd B, yield 46.2%.
Step 2:4-(4-phenyl-1,2,5-oxadiazole-2-oxide compound-3) methoxy benzyl alcohol (c 1) preparation
B0.33g (1.72mmo1) is dissolved in methylene dichloride l5mL, adds anhydrous pyridine 0.32mL (4mmol).In ice bath after cooling, instillation thionyl chloride 0.35ml (5.95mmol), stirred overnight at room temperature.By reaction solution frozen water (2 × 25ml) washing, be then washed till neutrality with saturated sodium bicarbonate aqueous solution, then wash with saturated aqueous common salt 25ml.Organic layer concentrates, and obtains orange-yellow oily matter.Be dissolved in 20ml acetonitrile by this oily matter, add p-Hydroxybenzylalcohol 647mg (5.2mmol), Anhydrous potassium carbonate 97mg (0.703mmol) and potassiumiodide 0.114g (0.687mmol), reacts 20h under room temperature.After reaction stops, filtering, filtrate concentrates, then with ether dissolution, elimination insolubles, with 5% sodium hydroxide solution (3 × 30mL) washing, organic layer is with anhydrous magnesium sulfate drying.Filter, filtrate concentrates, and silica gel column chromatography is separated, and with ethyl acetate: sherwood oil (60 ~ 90 DEG C)=1:4 ~ 1:3 (V:V) gradient elution, obtains faint yellow solid 0.32g, is compound c 1, productive rate 62.5%.
Step 3:4-[4-(2-oxygen-4-phenyl-1,2,5-oxadiazole-3 methoxyl group)-benzyloxy]-4-ketobutyric acid (d 1) preparation
Get c 1310mg(1.04mmol), Succinic anhydried 200mg(2mmol), DMAP is about 30mg(0.25mmol) dissolve with anhydrous methylene chloride 40ml, 40 DEG C of return stirring reaction 24h.After reaction terminates, respectively with hot water (2 × 20mL), saturated aqueous common salt (1 × 20ml) washs.Organic layer is with anhydrous sodium sulfate drying.After drying, filter, organic layer concentrates, and silica gel column chromatography is separated, and with methylene dichloride: methyl alcohol=50:0 ~ 50:1 gradient elution, obtaining pale yellow powder shape solid 400mg, is compound d 1, yield 96.7%.
Step 4:3-O-{4-[4-(2-oxygen-4-phenyl-1,2,5-oxadiazole-3 methoxyl group)-benzyloxy]-4-oxobutanoyl } preparation of-Brusatol (10)
By EDCI120mg(0.62mmol), intermediate d 1220mg(0.55mmol) be placed in reaction flask, in ice-water bath, add 30ml anhydrous methylene chloride, stir after 10 minutes, add Brusatol 160mg(0.31mmol) and DMAP70mg(0.57mmol), continue stirring reaction 3h under ice bath, then change with room temperature reaction 48h.Reaction solution respectively with rare HCl, rare NaHCO 3solution, saturated common salt water washing, organic layer is with anhydrous sodium sulfate drying.After drying, filter, filtrate concentrates, and silica gel column chromatography is separated, and with methylene dichloride: methyl alcohol=60:1 wash-out, obtains white powdery solids and be about 85mg, is compound 10, yield 30.7%.
The physical and chemical parameter of compound 10 is as follows:
1H NMR(500MHz,CD 3COCD 3)δ7.88(2H,dd,J=8.5，1.5Hz,ArH),7.61(3H,m,ArH),7.36(2H,d,J=8.5Hz,ArH),7.03(2H,d,J=8.5Hz,ArH),6.10(1H,br s,H-15),5.63(1H,s,H-2’),5.27(2H,s,OCH 2),5.08(2H,t,J=13.0Hz,OCH 2),4.96(1H,s,H-7),4.73(1H,d,J=7.5Hz,H a-20),4.26(2H,overlap,H-11,H-12),3.78(1H,d,J=7.5Hz,H b-20),3.70(3H,s,CH 3O-21),3.32(1H,br s,H-14),3.24(1H,d,J=12.5Hz,H-5),2.84(2H,m,COCH 2),2.81(1H,d,J=15.5Hz,H β-1),2.72(2H,t,J=6.0Hz,COCH 2),2.70(1H,overlap,H α-1),2.40(1H,br s,H-9),2.27(1H,dt,J=14.5,2.0Hz,H α-6),2.13(3H,s,H-5’),1.97(1H,ddd,J=14.0,12.5,2.0Hz,H β-6),1.91(3H,s,H-4’),1.77(3H,d,J=1.0Hz,H-18),1.50(3H,s,H-19);
13C NMR(125MHz,CD 3COCD 3)δ189.9(C-2),172.4(C=O),171.8(C-21),170.7(C=O),167.6(C-16),164.9(C-1’),159.0(C=N),158.3(ArC),158.1(C-3’),146.8(C-3),142.7(C-4),132.2(ArC),131.1(ArC),130.8(ArC×2),130.2(ArC×2),128.6(ArC×2),127.3(ArC),115.8(C-2’),115.8(ArC×2),113.1(C=N),83.2(C-7),82.3(C-13),76.4(C-12),74.0(C-20),72.7(C-11),67.7(C-15),66.3(OCH 2),59.6(OCH 2),52.7(C-21),51.0(C-14),50.4(C-1),46.1(C-8),43.2(C-5),41.6(C-9),41.5(C-10),29.5(CO CH 2,overlapped withCD3COCD3),29.4(CO CH 2),29.1(C-6),27.2(C-4’),20.2(C-5’),15.8(C-19),14.5(C-18)；
HRESIMS m/z923.2844[M+Na] +(calcd for C 46H 48N 2N aO 17,923.2845).
Embodiment 11:
3-O-{4-[3-(2-oxygen-4-phenyl-1,2,5-oxadiazole-3 methoxyl group)-benzyloxy]-4-oxobutanoyl }-Brusatol (11)
Step 1: with reference to step 2 in embodiment 10, by B first with thionyl chloride chloro, replaces p-Hydroxybenzylalcohol to be obtained by reacting 3-(4-phenyl-1,2,5-oxadiazole-2-oxide compound-3) methoxy benzyl alcohol (c with a salicylic alcohol 2).
Step 2: with reference to step 3 in embodiment 10, with c 2replace c 1, be obtained by reacting intermediate 4-[3-(2-oxygen-4-phenyl-1,2,5-oxadiazole-3 methoxyl group)-phenmethyl]-4-ketobutyric acid (d with Succinic anhydried 2).
Step 3: with reference to step 4 in embodiment 10, with d 2replace d 1, under the effect of excessive EDCI and DMAP, reacting obtained white powdery solids with Brusatol, is compound 11, yield 70.0%.
The physical and chemical parameter of compound 11 is as follows:
1H NMR(500MHz,CD 3COCD 3)δ7.88(2H,dd,J=8.0，1.5Hz,ArH),7.61(3H,m,ArH),7.31(1H,t,J=8.0Hz,ArH),7.06(2H,m,ArH),7.00(1H,dd,J=8.0,1.5Hz,ArH),6.11(1H,br s,H-15),5.63(1H,s,H-2’),5.27(2H,s,OCH 2),5.11(2H,t,J=13.0Hz,OCH 2),4.98(1H,s,H-7),4.73(1H,d,J=8.0Hz,H a-20),4.26(2H,overlap,H-11,H-12),3.78(1H,d,J=8.0Hz,H b-20),3.70(3H,s,CH 3O-21),3.33(1H,br s,H-14),3.23(1H,d,J=13.0Hz,H-5),2.86(2H,m,COCH 2),2.77(1H,overlap,H β-1),2.76(2H,t,J=6.0Hz,COCH 2),2.69(1H,J=16.0Hz,H α-1),2.39(1H,br s,H-9),2.27(1H,dt,J=14.5,2.0Hz,H α-6),2.14(3H,s,H-5’),1.96(1H,ddd,J=14.0,13.0,2.0Hz,H β-6),1.91(3H,s,H-4’),1.76(3H,s,H-18),1.48(3H,s,H-19);
13C NMR(125MHz,CD 3COCD 3)δ189.9(C-2),172.4(C=O),171.7(C-21),170.7(C=O),167.6(C-16),165.1(C-1’),159.0(C=N),158.4(ArC),158.3(C-3’),146.8(C-3),142.7(C-4),139.4(ArC),132.2(ArC),130.7(ArC),130.2(ArC×2),128.6(ArC×2),127.3(ArC),122.4(ArC),115.8(C-2’),115.4(ArC),115.2(ArC),113.1(C=N),83.2(C-7),82.3(C-13),76.5(C-12),74.0(C-20),72.7(C-11),67.6(C-15),66.3(OCH 2),59.6(OCH 2),52.7(C-21),51.0(C-14),50.4(C-1),46.1(C-8),43.2(C-5),41.6(C-9),41.5(C-10),29.5(CO CH 2,overlap-ped withCD 3COCD 3),29.4(CO CH 2),29.1(C-6),27.2(C-4’),20.2(C-5’),15.8(C-19),14.4(C-18)；
HRESIMS m/z923.2824[M+Na] +(calcd for C 46H 48N 2N aO 17,923.2845).
Embodiment 12:
3-O-{4-[2-(2-oxygen-4-phenyl-1,2,5-oxadiazole-3 methoxyl group)-benzyloxy]-4-oxobutanoyl }-Brusatol (12)
Step 1: with reference to step 2 in embodiment 10, by B first with thionyl chloride chloro, then replaces p-Hydroxybenzylalcohol to be obtained by reacting 2-(4-phenyl-1,2,5-oxadiazole-2-oxide compound-3) methoxy benzyl alcohol (c with diathesin 3).
Step 2: with reference to step 3 in embodiment 10, with c 3replace c 1, be obtained by reacting intermediate 4-[2-(2-oxygen-4-phenyl-1,2,5-oxadiazole-3 methoxyl group)-phenmethyl]-4-ketobutyric acid (d with Succinic anhydried 3).
Step 3: with reference to step 4 in embodiment 10, with d 3replace d 1, under the effect of excessive EDCI and DMAP, reacting obtained white powdery solids with Brusatol, is compound 12, yield 51.0%.
The physical and chemical parameter of compound 12 is as follows:
1H NMR(500MHz,CD 3COCD 3)δ7.89(2H,dd,J=7.5,1.5Hz,ArH),7.61(3H,m,ArH),7.35(2H,m,ArH),7.17(1H,d,J=7.5Hz,ArH),7.03(1H,t,J=7.5Hz，ArH),6.11(1H,br s,H-15),5.63(1H,s,H-2’),5.32(2H,s，OCH 2),5.04(2H,dd,J=12.5Hz，OCH 2),4.97(1H,s,H-7),4.73(1H,d,J=7.5Hz,H a-20),4.26(2H,overlap,H-11,H-12),3.78(1H,d,J=7.5Hz,H b-20),3.70(3H,s,CH 3O-21),3.33(1H,br s,H-14),3.22(1H,d,J=13.0Hz,H-5),2.82(2H,m,COCH 2),2.78(1H,overlap,H β-1),2.69(1H,overlap,H α-1),2.65(2H,t,J=6.5Hz,COCH 2),2.39(1H,br s,H-9),2.27(1H,dt,J=14.5,2.5Hz,H α-6),2.14(3H,s,H-5’),1.96(1H,ddd,J=14.0,12.5,2.5Hz,H β-6),1.91(3H,s,H-4’),1.75(3H,d,J=1.0Hz,H-18),1.49(3H,s,H-19);
13C NMR(125MHz,CD 3COCD 3)δ189.9(C-2),172.3(C=O),171.7(C-21),170.6(C=O),167.6(C-16),165.0(C-1’),159.0(C=N),158.2(C-3’),156.1(ArC),146.8(C-3),142.7(C-4),132.3(ArC),130.9(ArC),130.6(ArC),130.3(ArC×2),128.5(ArC×2),127.2(ArC),126.1(ArC),122.8(ArC),115.8(C-2’),113.1(C=N,ArC),83.2(C-7),82.3(C-13),76.5(C-12),74.0(C-20),72.7(C-11),67.6(C-15),61.9(OCH 2),59.8(OCH 2),52.7(C-21),51.1(C-14),50.4(C-1),46.1(C-8),43.2(C-5),41.6(C-9),41.5(C-10),29.5(CO CH 2),29.4(CO CH 2),29.1(C-6),27.2(C-4’),20.2(C-5’),15.8(C-19),14.4(C-18)；
HRESIMS m/z901.3027[M+H] +(calcd for C 46H 49N 2O 17,901.3026).
Embodiment 13:
3-O-[4-(2-oxygen-4-phenyl-1,2,5-oxadiazole-3 methoxyl group)-4-oxobutanoyl]-Brusatol (13)
Step 1: with reference to step 3 in embodiment 10, B is replaced c 1, direct and Succinic anhydried is obtained by reacting intermediate 4-(4-phenyl-1,2,5-oxadiazole-2-oxide compound-3) and methoxyl group-4-ketobutyric acid (d 4).
Step 2: with reference to step 4 in embodiment 10, with d 4replace d 1, under the effect of excessive EDCI and DMAP, reacting obtained white powdery solids with Brusatol, is compound 13, yield 41.8%.
The physical and chemical parameter of compound 13 is as follows:
1H NMR(500MHz,CD 3COCD 3)δ7.81(2H,dd,J=8.0，1.5Hz,ArH),7.61(3H,m,ArH),6.10(1H,br s,H-15),5.63(1H,s,H-2’),5.25(2H,t,J=14.0Hz,OCH 2),4.98(1H,s,H-7),4.74(1H,d,J=7.5Hz,H a-20),4.26(2H,s,H-11,H-12),3.78(1H,d,J=7.5Hz,H b-20),3.70(3H,s,CH 3O-21),3.33(1H,br s,H-14),3.25(1H,d,J=12.5Hz,H-5),2.84(2H,m,COCH 2),2.81(1H,d,J=15.5Hz,H β-1),2.68(1H,overlap,H α-1),2.65(2H,t,J=6.5Hz,COCH 2),2.39(1H,br s,H-9),2.27(1H,dt,J=14.5,2.0Hz,H α-6),2.14(3H,s,H-5’),1.96(1H,ddd,J=14.0,12.5,2.0Hz,H β-6),1.91(3H,s,H-4’),1.75(3H,s,H-18),1.49(3H,s,H-19);
13C NMR(125MHz,CD 3COCD 3)δ189.9(C-2),172.0(C=O),171.7(C-21),170.6(C=O),167.6(C-16),165.0(C-1’),159.0(C=N),158.1(C-3’),146.9(C-3),142.7(C-4),132.1(ArC),130.2(ArC×2),128.6(ArC×2),127.2(ArC),115.8(C-2’),112.5(C=N),83.2(C-7),82.3(C-13),76.4(C-12),74.0(C-20),72.7(C-11),67.6(C-15),55.3(OCH 2),52.7(C-21),51.0(C-14),50.4(C-1),46.1(C-8),43.2(C-5),41.6(C-9),41.5(C-10),29.4(CO CH 2),29.3(CO CH 2),29.0(C-6),27.2(C-4’),20.2(C-5’),15.8(C-19),14.4(C-18)；HRESIMS m/z817.2428[M+Na] +(calcd for C 39H 42N 2N aO 16,817.2427).
Embodiment 14:
3-O-[2-(2-oxygen-4-phenyl-1,2,5-oxadiazole-3 methoxyl group) carbonyl benzoyl base]-Brusatol (14)
Step 1: with reference to step 3 in embodiment 10, B is replaced c 1, direct and phthalic anhydride is obtained by reacting intermediate [2-(2-oxygen-4-phenyl-1,2,5-oxadiazole-3 methoxyl group)-carbonyl] phenylformic acid (d 5).
Step 2: with reference to step 4 in embodiment 10, with d 5replace d 1, under the effect of excessive EDCI and DMAP, reacting obtained white powdery solids with Brusatol, is compound 14, yield 30.5%.
The physical and chemical parameter of compound 14 is as follows:
1H NMR(500MHz,CD 3COCD 3)δ8.06(1H,m,ArH),7.81(2H,d,J=7.0Hz,ArH),7.72(3H,m,ArH),7.61(3H,m,ArH),6.14(1H,br s,H-15),5.64(1H,s,H-2’),5.52,5.41(each1H,d,J=13.5Hz,OCH 2),5.01(1H,s,H-7),4.76(1H,d,J=7.5Hz,H a-20),4.28(2H,overlap,H-11,H-12),3.81(1H,d,J=7.5Hz,H b-20),3.71(3H,s,CH 3O-21),3.35(1H,br s,H-14),3.27(1H,d,J=13.0Hz,H-5),2.85(1H,d,J=16.0Hz,H β-1),2.73(1H,d,J=16.0Hz,H α-1),2.42(1H,br s,H-9),2.30(1H,d,J=14.5Hz,H α-6),2.15(3H,s,H-5’),1.99(1H,ddd,J=14.0,13.0,1.5Hz,H β-6),1.92(3H,s,H-4’),1.82(3H,s,H-18),1.53(3H,s,H-19);
13C NMR(125MHz,CD 3COCD 3)δ189.7(C-2),171.7(C-21),167.7(C-16),167.5(C=O),165.0(C=O,C-1’),159.0(C=N),158.0(C-3’),147.7(C-3),142.6(C-4),133.1(ArC),132.7(ArC),132.5(ArC),132.2(ArC),131.1(ArC),130.8(ArC),130.2(ArC×2),129.8(ArC),128.6(ArC×2),127.2(ArC),115.8(C-2’),112.3(C=N),83.2(C-7),82.3(C-13),76.5(C-12),74.0(C-20),72.7(C-11),67.5(C-15),56.2(OCH 2),52.7(C-21),51.2(C-14),50.4(C-1),46.1(C-8),43.4(C-5),41.7(C-9),41.6(C-10),29.4(C-6),27.2(C-4’),20.3(C-5’),15.9(C-19),14.5(C-18)；
HRESIMS m/z843.2590[M+H] +(calcd for C 43H 43N 2O 16,843.2607).
Embodiment 15:
Synthetic route:
Step 1:4-(4-phenyl-1,2,5-oxadiazole-2-oxide compound-3)-methoxybenzyl bromine (e 1) preparation
Get c 1about 140mg(0.47mmol) be placed in reaction flask, dissolve with 20ml anhydrous methylene chloride.Under ice-water bath condition, add excessive PBr 3, continue to react in ice-water bath.After 2h, change and react 2h again with stirring at room temperature.After reaction terminates, in reaction solution, drip 1ml water, stir 10min.Reaction solution is with water (2 × 20ml) washing, and organic layer is with anhydrous sodium sulfate drying.After drying, filter, filtrate concentrates, and silica gel column chromatography is separated, with ethyl acetate: sherwood oil (60 ~ 90 DEG C)=1:8 (V:V) wash-out, obtains faint yellow solid 140mg, is Verbindung 1, productive rate 82.8%.
The preparation of step 2:3-O-[4-(2-oxygen-4-phenyl-1,2,5-oxadiazole-3)-methoxy-benzyl]-Brusatol (15)
By e 1about 150mg(0.42mmol), Brusatol is about 120mg(0.23mmol), Anhydrous potassium carbonate 70mg(0.51mmol) be placed in reaction flask, add anhydrous acetonitrile 20ml, 80 DEG C of return stirring 20h.Reaction solution cools, and filters, and filtrate concentrates, and dissolves with methylene dichloride 30ml.Wash with water (3 × 30ml) again.Water layer merges, and with 30ml dichloromethane extraction 1 time, merges all organic layers, with anhydrous sodium sulfate drying.After drying, filter, organic layer concentrates, and silica gel column chromatography is separated, and with methylene dichloride: methyl alcohol=50:1 (V:V) wash-out, obtains white powdery solids and be about 100mg, is compound 15, yield 54.2%.
The physical and chemical parameter of compound 15 is as follows:
1H NMR(500MHz,CD 3COCD 3)δ7.88(2H,dd,J=8.0,1.5Hz,ArH),7.60(3H,m,ArH),7.38(2H,d,J=8.5Hz,ArH),7.01(2H,d,J=8.5Hz,ArH),6.10(1H,br s,H-15),5.63(1H,s,H-2’),5.27(2H,s,OCH 2),4.93(1H,s,H-7),4.84,4.78(each1H,d,J=11.0Hz,OCH 2),4.71(1H,d,J=7.5Hz,H a-20),4.26(2H,overlap,H-11,H-12),3.75(1H,d,J=7.5Hz,H b-20),3.70(3H,s,CH 3O-21),3.30(1H,br s,H-14),3.09(1H,d,J=12.5Hz,H-5),2.78(1H,d,J=16.0Hz,H β-1),2.63(1H,d,J=16.0Hz,H α-1),2.31(1H,br s,H-9),2.22(1H,dt,J=14.5,2.0Hz,H α-6),2.13(3H,s,H-5’),1.91(3H,s,H-4’),1.84(1H,ddd,J=14.5,12.5,2.0Hz,H β-6),1.76(3H,d,J=1.5Hz,H-18),1.32(3H,s,H-19);
13C NMR(125MHz,CD 3COCD 3)δ193.4(C-2),171.7(C-21),167.6(C-16),165.0(C-1’),159.0(C-3’),158.2(C=N),158.0(ArC),146.1(C-3),144.9(C-4),132.7(ArC),132.2(ArC),131.2(ArC×2),130.2(ArC×2),128.5(ArC×2),127.2(ArC),115.8(C-2’),115.7(ArC×2),113.1(C=N),83.4(C-7),82.3(C-13),76.4(C-12),74.1(C-20),73.3(OCH 2),72.6(C-11),67.6(C-15),59.6(OCH 2),52.7(C-21),51.6(C-1),51.2(C-14),46.1(C-8),43.2(C-5),41.8(C-10),41.2(C-9),29.5(C-6,overlapped with CD3COCD3),27.2(C-4’),20.2(C-5’),15.7(C-19),14.4(C-18)；
HRESIMS m/z801.2867[M+H] +(calcd for C 42H 45N 2O 14,801.2865).
The synthetic route of compound 16-29 in embodiment:
The end product (compound numbers corresponds to the compound numbers in embodiment) of synthesis:
Compound R 6 X 1 Y 1
16 (CH 2) 3 O CO(CH 2) 2CO
17 (CH 2) 4 O CO(CH 2) 2CO
18 (CH 2) 2CH(CH 3) O CO(CH 2) 2CO
19 (CH 2) 5 O CO(CH 2) 2CO
20 (CH 2) 2O(CH 2) 2 O CO(CH 2) 2CO
21 CH 2C≡CCH 2 O CO(CH 2) 2CO
22 CH 2-p-Ph O CO(CH 2) 2CO
23 CH 2-m-Ph O CO(CH 2) 2CO
24 (CH 2) 4 O CO-o-PhCO
Compound R 7 X 2 Y 2
25 CH 2-p-Ph O CO(CH 2) 2CO
26 CH 2-m-Ph O CO(CH 2) 2CO
27 CH 2-o-Ph O CO(CH 2) 2CO
28 - O CO(CH 2) 2CO
29 - O CO-o-PhCO
Embodiment 16:
Step 1:3-O-(dimethyl tertiary butyl is silica-based)-Brusatol (C)
By Brusatol 110mg(0.21mmol), imidazoles 60mg(0.88mmol), DMAP is a small amount of, TERT-BUTYL DIMETHYL CHLORO SILANE 50mg(0.33mmol) and, dissolve with dry DMF 1.5ml, stirring reaction at 45 DEG C.After 3h, in reaction solution, add 20ml methylene dichloride, wash with water (2 × 30ml).Water layer extracts 1 time again with 20ml methylene dichloride, merges all organic layers, with anhydrous sodium sulfate drying.After drying, filter, filtrate concentrates, and silica gel column chromatography is separated, and with methylene dichloride: methyl alcohol=50:0 ~ 50:1 (V:V) wash-out, obtains faint yellow solid 120mg, is Compound C, yield 89.6%.
Step 2:3-O-(dimethyl tertiary butyl is silica-based)-bruceolide (D)
Get C110mg(0.17mmol) with 3ml dissolve with methanol, under ice-water bath condition, add the methanol solution 20ml of 1mol/L KOMe, continue at stirring reaction in ice-water bath.After 8h, in reaction solution, add Dowex50W-X2H +ion exchange resin, is adjusted to PH=7.Filter, filtrate concentrates, and changes and dissolves with chloroform.Organic layer is respectively with saturated sodium bicarbonate solution (1 × 30ml), and water (1 × 30ml), saturated aqueous common salt (1 × 30ml) washs.Organic layer is with anhydrous sodium sulfate drying, and after drying, concentrated, obtaining white solid and be about 49mg, is Compound D, yield 51.2%.
Step 3:15-O-{4-[3-(2-oxygen-3-benzenesulfonyl-1,2,5-oxadiazole-4-oxygen)-propoxy-]-4-oxobutanoyl }-bruceolide (16)
D is about 170mg(0.31mmol), EDCI110mg(0.57mmol), intermediate b 1220mg(0.55mmol), DMAP120mg(1mmol) be placed in reaction flask, in ice-water bath, add anhydrous methylene chloride dissolve, stir after 3h, change with room temperature reaction 48h.Reaction solution concentrates, and with tetrahydrofuran (THF) dilution, adds a small amount of Glacial acetic acid, adds the tetrahydrofuran solution of 1mol/LTBAF until desiliconization ether protecting group is complete in batches.Reaction stops, and adds 30ml methylene dichloride in reaction solution, more respectively with rare HCl(1 × 30ml), saturated NaHCO 3solution (1 × 30ml), saturated aqueous common salt (1 × 30ml) washs, and organic layer is with anhydrous sodium sulfate drying.After drying, filter, filtrate concentrates, and silica gel column chromatography is separated, and with methylene dichloride: methyl alcohol=50:1 wash-out, obtains white powdery solids and be about 98mg, is compound 16.Be that raw material calculates with Compound D, two step total recoverys 38.9%.
The physical and chemical parameter of compound 16 is as follows:
1H NMR(500MHz,CD 3COCD 3)δ8.07(2H,dd,J=8.0,1.0Hz,ArH),7.87(1H,t，J=7.5Hz)，7.41(2H,t,J=7.5Hz,ArH),6.81(1H,s，3-OH),6.14(1H,br s,H-15),4.93(1H,s,H-7),4.73(1H,d,J=8.0Hz,H a-20),4.55(2H,t，J=6.0Hz,OCH 2)，4.28(2H,t，J=6.0Hz,OCH 2),4.26(2H,overlap,H-11,H-12),3,79(3H,s,CH 3O-21),3.79(1H,overlap,H b-20),3.26(1H,d,J=12.0Hz,H-14),3.06(1H,d,J=13.0Hz,H-5),2.82(1H,J=16.0Hz,H β-1),2.69(1H,J=16.0Hz,H α-1),2.62(4H,s,COCH 2CH 2CO),2.36(1H,br s,H-9),2.24(1H,overlap,H α-6),2.22(2H,m,CH 2),1.89(1H,ddd,J=14.0,13.0,2.0Hz,H β-6),1.82(3H,d,J=1.5Hz,H-18),1.39(3H,s,H-19);
13C NMR(125MHz,CD 3COCD 3)d193.1(C-2),172.4(C=O),171.9(C=O),171.1(C-21),167.4(C-16),160.1(C=N),145.0(C-3),139.0(ArC),136.7(ArC),130.6(ArC×2),129.4(ArC×2),128.2(C-4),111.5(C=N),83.7(C-7),82.3(C-13),76.5(C-12),74.1(C-20),72.5(C-11),68.9(OCH 2),68.4(C-15),61.3(OCH 2),52.9(OCH 3),51.0(C-14),49.3(C-1),46.3(C-8),42.2(C-9),41.9(C-5),41.8(C-10),29.5(C-6),29.4(CO CH 2),29.3(CO CH 2),28.6(CH 2),15.6(C-19),13.3(C-18);
HRESIMS m/z843.1888[M+Na] +(calcd for C 36H 40N 2NaO 18S,843.1889).
Embodiment 17:
15-O-{4-[4-(2-oxygen-3-benzenesulfonyl-1,2,5-oxadiazole-4-oxygen)-butoxy]-4-oxobutanoyl }-bruceolide (17)
With reference to step 3 in embodiment 16, with b 2replace b 1, under the effect of excessive EDCI and DMAP, first react with D, then with TBAF desiliconization ether protecting group, obtained white powdery solids is compound 17, two step total recoverys 58.0%.
The physical and chemical parameter of compound 17 is as follows:
1H NMR(300MHz,CDCl 3)δ8.06(2H,br d,J=7.8Hz,ArH),7.77(1H,t，J=7.5Hz,ArH),7.63(2H,t,J=7.5Hz,ArH),6.38(1H,d,J=12.9Hz,H-15),6.08(1H,s，3-OH),4.73(1H,s,H-7),4.73(1H,d,J=7.5Hz,H a-20),4.46(2H,t，J=6.0Hz,OCH 2),4.25(1H,m,H-11),4.22(1H,s,H-12),4.19(2H,overlap,OCH 2),3.88(3H,s,CH 3O-21),3.81(1H,d,J=7.5Hz,H b-20),3.06(1H,d,J=13.2Hz,H-14),2.98(1H,J=16.2Hz,H β-1),2.93(1H,d,J=12.9Hz,H-5),2.68(4H,m,COCH 2CH 2CO),2.40(1H,J=16.5Hz,H α-1),2.35(1H,d,J=14.7Hz,H α-6),2.18(1H,d,J=3.3Hz,H-9),1.95(2H,m,CH 2),1.84(2H,m,CH 2),1.84(3H,s,H-18),1.77(1H,ddd,J=14.7,12.9,2.4Hz,H β-6),1.39(3H,s,H-19);
13C NMR(75MHz,CDCl 3)δ192.1(C-2),172.3(C=O),171.8(C=O),170.6(C-21),166.7(C-16),158.9(C=N),144.0(C-3),137.9(ArC),135.6(ArC),129.6(ArC×2),128.5(ArC×2),127.8(C-4),110.4(C=N),82.8(C-7),81.4(C-13),75.5(C-12),73.9(C-20),70.9(C-11,OCH 2),66.9(C-15),64.1(OCH 2),53.1(OCH 3),51.5(C-14),48.4(C-1),45.6(C-8),41.9(C-9),41.8(C-5),41.0(C-10),.28.9(C-6),28.7(CO CH 2),28.3(CO CH 2),25.1(CH 2),24.8(CH 2),15.3(C-19),13.2(C-18);
HRESIMS m/z857.2050[M+Na] +(calcd for C 37H 42N 2NaO 18S,857.2046).
Embodiment 18:
(±)-15-O-{4-[(2-oxygen-3-benzenesulfonyl-1,2,5-oxadiazole-4-oxygen)-3-butoxy]-4-oxobutanoyl]-4-oxobutanoyl }-bruceolide (18)
With reference to step 3 in embodiment 16, with b 3replace b 1, under the effect of excessive EDCI and DMAP, first react with D, then with TBAF desiliconization ether protecting group, obtained white powdery solids is compound 18, two step total recoverys 47.8%.
The physical and chemical parameter of compound 18 is as follows:
1h NMR (500MHz, CDCl 3) (be raceme, part hydrogen display m peak, therefore peak shape is not indicated) δ 8.05 (2H, ArH), 7.75 (1H, ArH), 7.62 (2H, ArH), 6.31 (1H, H-15), 6.11 (1H, 3-OH), 5.16 (1H, OCH), 4.71 (1H, H-7), 4.73 (1H, H a-20), 4.45(2H, OCH 2), 4.24(1H, H-11), 4.20 (1H, s, H-12), 4.10,3.78(each1H, OCH 2), 3.84 (3H, s, CH 3o-21), 3,46 (1H, H b-20), 3.03 (1H, H-14), 2.96 (1H, H β-1), 2.91 (1H, H-5), 2.60 (4H, COCH 2cH 2cO), 2.42 (1H, H α-1), 2.35 (1H, H α-6), 2.17 (1H, H-9), 2.10 (2H, CH 2), 1.83 (3H, H-18), 1.75 (1H, H β-6), 1.37 (3H, H-19), 1.32 (3H, CH 3)
13C NMR(125MHz,CDCl 3)δ192.1(C-2),171.8(C=O),171.7(C=O),170.6(C-21),166.7(C-16),158.9(C=N),144.0(C-3),137.9(ArC),135.6(ArC),129.6(ArC×2),128.6(ArC×2),127.7(C-4),110.6(C=N),82.8(C-7),81.4(C-13),75.5(C-12),74.0(C-20),70.9(C-11),68.1(OCH),67.7(OCH 2),66.9(C-15),53.1(OCH 3),51.7(C-14),48.5(C-1),45.7(C-8),41.9(C-9),41.8(C-5),41.0(C-10),34.7(CH 2),29.0(C-6),29.0(CO CH 2),28.4(CO CH 2),20.1(CH 3),15.4(C-19),13.3(C-18);
HRESIMS m/z857.2058[M+Na] +(calcd for C 37H 42N 2NaO 18S,857.2046).
Embodiment 19:
15-O-{4-[5-(2-oxygen-3-benzenesulfonyl-1,2,5-oxadiazole-4-oxygen)-pentyloxy]-4-oxobutanoyl }-bruceolide (19)
With reference to step 3 in embodiment 16, with b 4replace b 1, under the effect of excessive EDCI and DMAP, first react with D, then with TBAF desiliconization ether protecting group, obtained white powdery solids is compound 19, two step total recoverys 38.6%.
The physical and chemical parameter of compound 19 is as follows:
1H NMR(500MHz,CDCl 3)δ8.04(2H,br d,J=7.5Hz,ArH),7.76(1H,t，J=7.5Hz,ArH),7.62(2H,t,J=7.5Hz,ArH),6.38(1H,d,J=12.0Hz,H-15),6.09(1H,s，3-OH),4.73(1H,s,H-7),4.72(1H,d,J=8.0Hz,H a-20),4.42(2H,t，J=6.0Hz,OCH 2),4.24(1H,br s,H-11)，4.22(1H,s,H-12),4.19（2H,m,OCH 2），3.86(3H,s,CH 3O-21),3.79(1H,d,J=8.0Hz,H b-20),3.06(1H,d,J=13.0Hz,H-14),2.96(1H,J=16.5Hz,H β-1),2.92(1H,d,J=13.5Hz,H-5),2.66(4H,m,COCH 2CH 2CO),2.40(1H,J=16.5Hz,H α-1),2.35(1H,dt,J=14.5,2.0Hz,H α-6),2.18(1H,br s,H-9),1.90(2H,m,CH 2),1.83(3H,s,H-18),1.76(1H,ddd,J=14.5,13.5,2.0Hz,H β-6),1.72(2H,m,CH 2),1.54(2H,m,CH 2),1.38(3H,s,H-19);
13C NMR(125MHz,CDCl 3)δ192.0(C-2),172.5(C=O),171.6(C=O),170.6(C-21),166.7(C-16),159.0(C=N),144.0(C-3),138.0(ArC),135.6(ArC),129.6(ArC×2),128.5(ArC×2),127.5(C-4),110.5(C=N),82.8(C-7),81.5(C-13),75.6(C-12),74.1(C-20),71.2(OCH 2),71.0(C-11,),66.8(C-15),64.6(OCH 2),53.2(OCH 3),51.7(C-14),48.5(C-1),45.8(C-8),42.2(C-9),41.9(C-5),41.1(C-10),.29.0(C-6),28.7(CO CH 2),28.4(CO CH 2),28.0(CH 2×2),22.1(CH 2),15.4(C-19),13.3(C-18);
HRESIMS m/z849.2383[M+H] +(calcd for C 38H 45N 2O 18S,849.2383).
Embodiment 20:
15-O-{4-{2-{2-[(2-oxygen 3-benzenesulfonyl-1,2,5-oxadiazole-4-oxygen) oxyethyl group] }-oxyethyl group }-4-oxobutanoyl }-bruceolide (20)
With reference to step 3 in embodiment 16, with b 5replace b 1, under the effect of excessive EDCI and DMAP, first react with D, then with TBAF desiliconization ether protecting group, obtained white powdery solids is compound 20,
Two step total recoverys 48.7%.
The physical and chemical parameter of compound 20 is as follows:
1H NMR(300MHz,CDCl 3)δ8.05(2H,br d,J=7.5Hz,ArH),7.75(1H,t，J=7.5Hz,ArH),7.61(2H,t,J=7.5Hz,ArH),6.34(1H,d,J=13.2Hz,H-15),6.10(1H,s，3-OH),4.73(1H,s,H-7),4.71(1H,d,J=8.1Hz,H a-20),4.56(2H,t,J=3.9Hz,OCH 2),4.29(2H,m,OCH 2），4.23(1H,d,J=3.3Hz H-11),4.20(1H,s,H-12),3.90(2H,t，J=4.2Hz,OCH 2),3.84(3H,s,CH 3O-21),3.79(1H,overlap,H b-20),3.79(2H,t，J=4.2Hz,OCH 2),3.06(1H,d,J=13.2Hz,H-14),2.95(1H,J=15.9Hz,H β-1),2.92(1H,d,J=12.6Hz,H-5),2.68(4H,m,COCH 2CH 2CO),2.40(1H,J=15.9Hz,H α-1),2.35(1H,d,J=14.4Hz,H α-6),2.18(1H,br s,H-9),1.82(3H,s,H-18),1.75(1H,ddd,J=14.4,12.6,2.4Hz,H β-6),1.37(3H,s,H-19);
13C NMR(75MHz,CDCl 3)δ192.0(C-2),172.3(C=O),171.6(C=O),170.5(C-21),166.7(C-16),158.9(C=N),144.0(C-3),137.9(ArC),135.6(ArC),129.6(ArC×2),128.5(ArC×2),127.6(C-4),110.5(C=N),82.8(C-7),81.5(C-13),75.5(C-12),74.0(C-20),71.0(C-11),70.5（OCH 2），69.2（OCH 2）,68.3（OCH 2），66.8(C-15),63.8(OCH 2),53.1(OCH 3),51.6(C-14),48.4(C-1),45.7(C-8),42.1(C-9),41.9(C-5),41.0(C-10),29.0(C-6),28.7(CO CH 2),28.3(CO CH 2),15.3(C-19),13.3(C-18);
HRESIMS m/z873.2000[M+Na] +(calcd for C 37H 42N 2NaO 19S,873.1995).
Embodiment 21:
15-O-{4-[4-(2-oxygen-3-benzenesulfonyl-1,2,5-oxadiazole-4-oxygen)-2-alkynes-butoxy]-4-oxobutanoyl }-bruceolide (21)
With reference to step 3 in embodiment 16, with b 6replace b 1, under the effect of excessive EDCI and DMAP, first react with D, then with TBAF desiliconization ether protecting group, obtained white powdery solids is compound 21, two step total recoverys 33.7%.
The physical and chemical parameter of compound 21 is as follows:
1H NMR(500MHz,CDCl 3)δ8.06(2H,br d,J=7.5Hz,ArH),7.76(1H,t，J=7.5Hz),7.63(2H,t,J=7.5Hz,ArH),6.36(1H,d,J=11.0Hz,H-15),6.10（1H,s，3-OH）,5.10（2H,s，OCH 2）,4.77(1H,s,H-7),4.76(2H,overlap,OCH 2),4.73(1H,d,J=8.0Hz,H a-20),4.25（1H,d,J=3.5Hz,H-11），4.22(1H,s,H-12),3.86(3H,s,CH 3O-21),3.79(1H,d,J=8.0Hz,H b-20),3.06(1H,d,J=13.0Hz,H-14),2.96(1H,J=16.0Hz,H β-1),2.93(1H,d,J=12.5Hz,H-5),2.70(4H,m,COCH 2CH 2CO),2.41(1H,J=16.0Hz,H α-1),2.36(1H,dt,J=15.0,2.5Hz,H α-6),2.19(1H,d,J=3.0Hz,H-9),1.83(3H,d,J=1.5Hz,H-18),1.76(1H,ddd,J=14.0,12.5,2.5Hz,H β-6),1.38(3H,s,H-19);
13C NMR(125MHz,CDCl 3)δ192.1(C-2),171.7(C=O),171.5(C=O),170.4(C-21),166.7(C-16),157.9(C=N),144.0(C-3),137.7(ArC),135.7(ArC),129.7(ArC×2),128.6(ArC×2),127.7(C-4),110.6(C=N),83.6(C≡C,1C),82.8(C-7),81.4(C-13),78.8(C≡C,1C),75.6(C-12),74.0(C-20),71.0(C-11),66.9(C-15),58.6(OCH 2),53.2(OCH 3),52.3(OCH 2),51.6(C-14),48.5(C-1),45.7(C-8),42.1(C-9),41.9(C-5),41.0(C-10),29.0(C-6),28.5(CO CH 2),28.3(CO CH 2),15.4(C-19),13.3(C-18);
HRESIMS m/z853.1750[M+Na] +(calcd for C 37H 38N 2NaO 18S,853.1733).
Embodiment 22:
15-O-{4-[4-(2-oxygen-3-benzenesulfonyl-1,2,5-oxadiazole-4 oxygen)-benzyloxy]-4-oxobutanoyl }--bruceolide (22)
With reference to step 3 in embodiment 16, with b 7replace b 1, under the effect of excessive EDCI and DMAP, first react with D, then with TBAF desiliconization ether protecting group, obtained white powdery solids is compound 22,
Two step total recoverys 35.3%.
The physical and chemical parameter of compound 22 is as follows:
1H NMR(500MHz,CD 3COCD 3)δ8.08(2H,br d,J=7.5Hz,ArH),7.90(1H,t，J=7.5Hz，ArH),7.76(2H,t,J=7.5Hz,ArH),7.54(2H,d,J=8.5Hz,ArH),7.43(2H,d,J=8.5Hz,ArH),6.83（1H,s，3-OH）,6.16(1H,br s,H-15),5.19(each1H,d,J=13.0Hz,OCH 2),4.96(1H,s,H-7),4.73(1H,d,J=8.0Hz,H a-20),4.26(2H,overlap,H-11,H-12),3.78(1H,overlap,H b-20),3.77(3H,s,CH 3O-21),3.27(1H,d,J=12.5Hz,H-14),3.12(1H,d,J=13.0Hz,H-5),2.83(1H,J=16.0Hz,H β-1),2.71(1H,J=16.0Hz,H α-1),2.65(4H,m,COCH 2CH 2CO),2.40(1H,br s,H-9),2.26(1H,dt,J=15.0,2.5Hz,H α-6),1.90(1H,ddd,J=14.5,13.0,2.5Hz,H β-6),1.82(3H,d,J=1.5Hz,H-18),1.39(3H,s,H-19);
13C NMR(125MHz,CD 3COCD 3)δ193.2(C-2),172.3(C=O),171.9(C=O),171.1(C-21),167.4(C-16),159.6(C=N),153.6（ArC）145.1(C-3),138.8(ArC),136.8(ArC),135.9(ArC),130.7(ArC×2),130.7(ArC×2),129.5(ArC×2),128.1(C-4),120.6(ArC×2),112.1(C=N),83.7(C-7),82.4(C-13),76.5(C-12),74.1(C-20),72.6(C-11),68.5(C-15),65.9(OCH 2),52.9(OCH 3),51.2(C-14),49.3(C-1),46.4(C-8),42.3(C-9),41.9(C-5),41.9(C-10),29.7(C-6).29.5(CO CH 2),29.5（CO CH 2,overlapped withCD 3COCD 3）,15.6(C-19),13.3(C-18);
HRESIMS m/z891.1902[M+Na] +(calcd for C 40H 40N 2NaO 18S,891.1889).
Embodiment 23:
15-O-{4-[3-(2-oxygen-3-benzenesulfonyl-1,2,5-oxadiazole-4 oxygen)-benzyloxy]-4-oxobutanoyl }-bruceolide (23)
With reference to step 3 in embodiment 16, with b 8replace b 1, under the effect of excessive EDCI and DMAP, first react with D, then with TBAF desiliconization ether protecting group, obtained white powdery solids is compound 23,
Two step total recoverys 29.7%.
The physical and chemical parameter of compound 23 is as follows:
1H NMR(500MHz,CD 3COCD 3)δ8.08(2H,dd,J=8.5,1.5Hz,ArH),7.90(1H,t，J=7.5Hz，ArH),7.75(2H,t,J=8.0Hz,ArH),7.50(1H,t,J=8.0Hz,ArH),7.46(1H,t,J=1.5Hz,ArH),7.39(2H,m,ArH),6.84（1H,s，3-OH）,6.16(1H,br s,H-15),5.17(2H,s,OCH 2),4.96(1H,t,J=2.5Hz,H-7),4.73(1H,d,J=7.5Hz,H a-20),4.25(2H,overlap,H-11,H-12),3.78(1H,overlap,H b-20),3.77(3H,s,CH3O-21),3.27(1H,d,J=12.5Hz,H-14),3.10(1H,d,J=13.0Hz,H-5),2.83(1H,J=16.5Hz,H β-1),2.71(1H,overlap,H α-1),2.66(4H,m,COCH 2CH 2CO),2.39(1H,br s,H-9),2.25(1H,dt,J=15.0,2.5Hz,H α-6),1.89(1H,ddd,J=14.0,13.0,2.5Hz,H β-6),1.82(3H,d,J=1.5Hz,H-18),1.39(3H,s,H-19);
13C NMR(125MHz,CD 3COCD 3)δ193.2(C-2),172.2(C=O),171.9(C=O),171.0(C-21),167.5(C-16),159.6(C=N),154.1(ArC),145.0(C-3),139.9(ArC),138.8(ArC),136.8(ArC),131.0(ArC),130.7(ArC×2),129.5(ArC×2),128.2(C-4),126.7(ArC),120.1(ArC),119.8(ArC),112.1(C=N),83.7(C-7),82.4(C-13),76.5(C-12),74.1(C-20),72.5(C-11),68.5(C-15),65.8(OCH 2),52.9(OCH 3),51.1(C-14),49.3(C-1),46.3(C-8),42.2(C-9),41.9(C-5),41.9(C-10),29.6(C-6,overlapped withCD 3COCD 3).29.5(CO CH 2),29.5（CO CH 2,overlapped with CD 3COCD 3）,15.6(C-19),13.3(C-18);
HRESIMS m/z891.1898[M+Na] +(calcd for C 40H 40N 2NaO 18S,891.1889).
Embodiment 24:
15-O-{2-[4-(2-oxygen-3-benzenesulfonyl-1,2,5-oxadiazole-4-oxygen)-butoxy carbonyl]-benzoyl }-bruceolide (24)
With reference to step 3 in embodiment 16, with b 9replace b 1, under the effect of excessive EDCI and DMAP, first react with D, then with TBAF desiliconization ether protecting group, obtained white powdery solids is compound 24, two step total recoverys 27.8%.
The physical and chemical parameter of compound 24 is as follows:
1H NMR(500MHz,CDCl 3)δ8.04(2H,br d,J=7.8Hz,ArH),7.74(3H,m，ArH),7.58(4H,m,ArH),6.62(1H,d,J=11.0Hz,H-15),6.10（1H,s，3-OH）,4.77(1H,s,H-7),4.73(1H,d,J=8.0Hz,Ha-20),4.48（2H,t，J=6.0Hz,OCH 2）,4.43(2H,m,OCH 2),4.30(1H,d,J=4.0Hz,H-11),4.26(1H,s,H-12),3.80(1H,d,J=7.5Hz,H b-20),3.43(3H,s,CH 3O-21),3.18(1H,d,J=13.5Hz,H-14),2.99(1H,J=16.0Hz,H β-1),2.96(1H,overlap,H-5),2.43(1H,J=16.0Hz,H α-1),2.40(1H,overlap,H α-6),2.26(1H,d,J=3.3Hz,H-9),2.00(4H,m,CH 2),1.84(3H,s,H-18),1.78(1H,ddd,J=14.5,13.0,2.5Hz,H β-6),1.39(3H,s,H-19);
13C NMR(125MHz,CDCl 3)δ192.3(C-2),171.2(C-21),167.3(C-16),166.8(C=O),165.5(C=O),159.2(C=N),144.3(C-3),138.2(ArC),135.9(ArC),131.7(ArC×2),129.9(ArC×2),129.2(ArC×2),129.1(ArC×2)128.8(ArC×2),127.8(C-4),110.7(C=N),83.2(C-7),81.9(C-13),75.6(C-12),74.4(C-20),71.3(C-11),71.2(OCH 2)，67.9(C-15),65.5(OCH 2),53.0(OCH 3),52.0(C-14),48.7(C-1),46.2(C-8),42.4(C-9),42.2(C-5),41.3(C-10),29.3(C-6),25.5(CH 2),25.1(CH 2),15.6(C-19),13.6(C-18);
HRESIMS m/z905.2057[M+Na] +(calcd for C 41H 42N 2NaO 18S,905.2046).
Embodiment 25:
15-O-{4-[4-(2-oxygen-4-phenyl-1,2,5-oxadiazole-3 methoxyl group)-benzyloxy]-4-oxobutanoyl }-bruceolide (25)
With reference to step 3 in embodiment 16, with d 1replace b 1, under the effect of excessive EDCI and DMAP, first react with D, then with TBAF desiliconization ether protecting group, obtained white powdery solids is compound 25, two step total recoverys 56.4%.
The physical and chemical parameter of compound 25 is as follows:
1H NMR(300MHz,CDCl 3)δ7.83(2H,dd,J=7.5,1.2Hz,ArH),7.53(3H,m,ArH)，7.31(2H,d,J=7.8Hz,ArH),6.97(2H,d,J=7.8Hz,ArH),6.36(1H,d,J=12.9Hz,H-15),6.12(1H,s，3-OH),5.09(2H,s,OCH 2),5.07(2H,s,OCH 2),4.74(1H,s,H-7),4.73(1H,d,J=8.1Hz,H a-20),4.25（1H,d,J=3.9Hz,H-11），4.22(1H,s,H-12),3.84(3H,s,CH 3O-21),3.79(1H,d,J=8.1Hz,H b-20),3.06(1H,d,J=13.2Hz,H-14),2.96(1H,J=16.2Hz,H β-1),2.91(1H,overlap,H-5),2.67(4H,m,COCH 2CH 2CO),2.41(1H,J=16.2Hz,H α-1),2.35(1H,d,J=14.1Hz,H α-6),2.18(1H,d,J=3.0Hz,H-9),1.83(3H,s,H-18),1.76(1H,ddd,J=14.1,13.2,1.5Hz,H β-6),1.38(3H,s,H-19);
13C NMR(75MHz,CDCl 3)δ192.1(C-2),172.2(C=O),171.6(C=O),170.6(C-21),166.8(C-16),157.0(C=N),156.9(ArC),144.0(C-3),131.4(ArC),130.3(ArC×2),129.5(ArC),129.3(ArC×2),127.7(C-4),127.6(ArC×2),126.0(ArC),114.9(ArC×2),112.0(C=N),82.8(C-7),81.4(C-13),75.5(C-12),74.0(C-20),70.9(C-11),66.9(C-15),66.3(OCH 2),58.3(OCH 2),53.1(OCH 3),51.6(C-14),48.4(C-1),45.7(C-8),42.0(C-9),41.9(C-5),41.0(C-10),29.0(C-6),28.8(CO CH 2),28.3(CO CH 2),15.3(C-19),13.3(C-18)
HRESIMS m/z841.2436[M+Na] +(calcd for C 41H 42N 2NaO 16,841.2427).
Embodiment 26:
15-O-{4-[3-(2-oxygen-4-phenyl-1,2,5-oxadiazole-3 methoxyl group)-benzyloxy]-4-oxobutanoyl }-bruceolide (26)
With reference to step 3 in embodiment 16, with d 2replace b 1, under the effect of excessive EDCI and DMAP, first react with D, then with TBAF desiliconization ether protecting group, obtained white powdery solids is compound 26, two step total recoverys 31.5%.
The physical and chemical parameter of compound 26 is as follows:
1H NMR(300MHz,CDCl 3)δ7.83(2H,dd,J=7.8,1.8Hz,ArH),7.53(3H,m,ArH),7.29(1H,t,J=8.1Hz,ArH),6.96(3H,m,ArH),6.39(1H,d,J=13.2Hz,H-15),6.10(1H,s，3-OH),5.16,5.11(each1H,d,J=12.3Hz,OCH 2),5.15,5.09(each1H,d,J=12.3Hz,OCH 2),4.72(1H,s,H-7),4.72(1H,d,J=7.8Hz,Ha-20),4.24(1H,br s,H-11),4.20(1H,s,H-12),3.84(3H,s,CH 3O-21),3.79(1H,d,J=7.8Hz,H b-20),3.05(1H,d,J=13.2Hz,H-14),2.96(1H,J=16.2Hz,H β-1),2.96(1H,overlap,H-5),2.73(4H,m,COCH 2CH 2CO),2.41(1H,J=16.2Hz,H α-1),2.36(1H,dt,J=14.1,2.1Hz,H α-6),2.18(1H,d,J=3.6Hz,H-9),1.84(3H,s,H-18),1.76(1H,ddd,J=14.1,12.9,2.1Hz,H β-6),1.38(3H,s,H-19);
13C NMR(75MHz,CDCl 3)δ192.0(C-2),172.2(C=O),171.7(C=O),170.6(C-21),166.7(C-16),157.2(ArC),157.1(C=N),144.0(C-3),137.7(ArC),131.4(ArC),129.9(ArC),129.3(ArC×2),127.7(ArC×2),127.5(C-4),126.1(ArC),121.7(ArC),115.1(ArC),113.8(ArC),112.2(C=N),82.8(C-7),81.5(C-13),75.6(C-12),74.1(C-20),71.0(C-11),66.9(C-15),66.2(OCH 2),58.4(OCH 2),53.2(OCH 3),51.8(C-14),48.4(C-1),45.7(C-8),42.0(C-9),41.8(C-5),41.0(C-10),29.0(C-6),28.8(CO CH 2),28.4(COCH 2),15.4(C-19),13.3(C-18)
HRESIMS m/z841.2435[M+Na] +(calcd for C 41H 42N 2NaO 16,841.2427).
Embodiment 27:
15-O-{4-[2-(2-oxygen-4-phenyl-1,2,5-oxadiazole-3 methoxyl group)-benzyloxy]-4-oxobutanoyl }-bruceolide (27)
With reference to step 3 in embodiment 16, with d 3replace b 1, under the effect of excessive EDCI and DMAP, first react with D, then with TBAF desiliconization ether protecting group, obtained white powdery solids is compound 27, two step total recoverys 70.2%.
The physical and chemical parameter of compound 27 is as follows:
1H NMR(300MHz,CDCl 3)δ7.83(2H,d,J=7.8Hz,ArH),7.53(3H,m,ArH),7.32(2H,m,ArH),7.02(2H,t,J=8.7Hz,ArH),6.33(1H,d,J=12.9Hz,H-15),6.12（1H,s，3-OH）,5.13(2H,s,OCH 2),5.05(2H,s,OCH 2),4.72(1H,s,H-7),4.72(1H,overlap,H a-20),4.23（1H,br s,H-11），4.19(1H,s,H-12),3.81(3H,s,CH 3O-21),3.79(1H,overlap,H b-20),3.05(1H,d,J=13.2Hz,H-14),2.95(1H,J=15.9Hz,H β-1),2.91(1H,overlap,H-5),2.60(4H,m,COCH 2CH 2CO),2.41(1H,J=15.9Hz,H α-1),2.36(1H,d,J=14.1Hz,H α-6),2.18(1H,br s,H-9),1.82(3H,s,H-18),1.75(1H,ddd,J=14.1,12.9,2.4Hz,H β-6),1.38(3H,s,H-19);
13C NMR(75MHz,CDCl 3)δ192.2(C-2),172.1(C=O),171.5(C=O),170.5(C-21),166.8(C-16),157.0(C=N),155.0(ArC),144.0(C-3),131.4(ArC),130.6(ArC),130.1(ArC),129.4(ArC×2),127.8(C-4),127.6(ArC×2),126.0(ArC),124.5(ArC),122.2(ArC),112.0(C=N),111.9(ArC),82.7(C-7),81.4(C-13),75.5(C-12),73.9(C-20),70.9(C-11),66.8(C-15),61.9(OCH 2),58.6(OCH 2),53.1(OCH 3),51.6(C-14),48.4(C-1),45.7(C-8),42.0(C-9),41.8(C-5),41.0(C-10),29.0(C-6),28.6(CO CH 2),28.3(CO CH 2),15.3(C-19),13.3(C-18)
HRESIMS m/z857.2162[M+K] +(calcd for C 41H 42KN 2O 16,857.2166).
Embodiment 28:
15-O-[4-(2-oxygen-4-phenyl-1,2,5-oxadiazole-3 oxygen)-4-oxobutanoyl]-bruceolide (28)
With reference to step 3 in embodiment 16, with d 4replace b 1, under the effect of excessive EDCI and DMAP, first react with D, then with TBAF desiliconization ether protecting group, obtained white powdery solids is compound 28, two step total recoverys 24.0%.
The physical and chemical parameter of compound 28 is as follows:
1H NMR(500MHz,CD 3COCD 3)δ7.83(2H,m,ArH),7.62(3H,m,ArH)，6.82(1H,s，3-OH),6.14(1H,br s,H-15),5.26,5.24(each1H,d,J=14.0Hz,OCH 2),4.94(1H,s,H-7),4.72(1H,d,J=7.5Hz,H a-20),4.25(2H,overlap,H-11,H-12),3.76(1H,overlap,Hb-20),3.75(3H,s,CH 3O-21),3.27(1H,d,J=12.5Hz,H-14),3.11(1H,d,J=13.0Hz,H-5),2.83(1H,J=16.0Hz,H β-1),2.71(1H,J=16.0Hz,H α-1),2.61(4H,m,COCH 2CH 2CO),2.38(1H,br s,H-9),2.25(1H,dt,J=14.7,2.5Hz,H α-6),1.90(1H,ddd,J=14.5,12.5,2.5Hz,H β-6),1.82(3H,s,H-18),1.39(3H,s,H-19);
13C NMR(125MHz,CD 3COCD 3)δ193.2(C-2),172.0(C=O),171.9(C=O),171.0(C-21),167.4(C-16),158.1(C=N),145.1(C-3),132.1(ArC),130.2(ArC×2),128.7(ArC×2),128.1(C-4),127.2(ArC),112.6(C=N),83.7(C-7),82.4(C-13),76.5(C-12),74.1(C-20),72.6(C-11),68.6(C-15),55.2(OCH 2),52.9(OCH 3),51.1(C-14),49.3(C-1),46.3(C-8),42.3(C-9),41.9(C-5),41.9(C-10),29.7（C-6）,29.2(CO CH 2),29.2(CO CH 2),15.6(C-19),13.3(C-18)
HRESIMS m/z713.2172[M+H] +(calcd for C 34H 37N 2O 15,713.2188).
Embodiment 29:
15-O-[2-(2-oxygen-4-phenyl-1,2,5-oxadiazole-3 methoxyl group) carbonyl benzoyl base]-bruceolide (29)
With reference to step 3 in embodiment 16, with d 5replace b 1, under the effect of excessive EDCI and DMAP, first react with D, then with TBAF desiliconization ether protecting group, obtained white powdery solids is compound 29, two step total recoverys 17.8%.
The physical and chemical parameter of compound 29 is as follows:
1H NMR(500MHz,CDCl 3)δ7.80(1H,dd,J=7.5,0.5Hz,ArH),7.76(2H,dd,J=7.5,0.5Hz,ArH),7.71(1H,dd,J=7.5,0.5Hz,ArH),7.59(5H,m,ArH),6.59(1H,d,J=13.2Hz,H-15),6.09(1H,s，3-OH),5.49,5.38(each1H,d,J=13.0Hz,OCH 2),4.79(1H,s,H-7),4.74(1H,d,J=8.0Hz,H a-20),4.30(1H,m,H-11)，4.24(1H,s,H-12),3.84(1H,d,J=7.8Hz,H b-20),3.42(3H,s,CH 3O-21),3.18(1H,d,J=12.5Hz,H-14),3.01(1H,J=16.5Hz,H β-1),2.96(1H,d,J=13.0Hz,H-5),2.42(1H,J=16.0Hz,H α-1),2.42(1H,overlap,H α-6),2.23(1H,br s,H-9),1.87(3H,s,H-18),1.79(1H,ddd,J=14.0,13.0,2.5Hz,H β-6),1.42(3H,s,H-19);
13C NMR(125MHz,CDCl 3)δ191.9(C-2),171.1(C-21),168.1(C-16),166.4(C=O),165.0(C=O),156.8(C=N),144.1(C-3),132.1(ArC×2),131.7(ArC×2),131.5(ArC),129.5(ArC×2),129.3(ArC×2),127.7(ArC×2),127.3(C-4),125.9(ArC),111.3(C=N),82.9(C-7),81.6(C-13),75.4(C-12),74.1(C-20),71.1(C-11),67.8(C-15),55.4(OCH 2),52.8(OCH 3),51.7(C-14),48.5(C-1),45.9(C-8),42.3(C-9),42.0(C-5),41.1(C-10),29.1(C-6),15.4(C-19),13.3(C-18)
HRESIMS m/z783.2014[M+Na] +(calcd for C 38H 36N 2NaO 15,783.2008).
Embodiment 30:
Synthetic route:
Step 1:3-(2-oxygen-3-benzenesulfonyl-1,2,5-oxadiazole-4-) oxygen iodopropane (g 1)
Get a 185mg(0.28mmol), triphenylphosphine is about 90mg(0.34mmol), imidazoles 40mg(0.61mmol) dissolve with anhydrous methylene chloride 40ml.After dissolving, add I 2about 90mg(0.35mmol), room temperature lucifuge stirring reaction.After 5h, reaction solution is respectively with rare NaHCO 3solution, 10%Na 2s 2o 3solution washing, organic layer is with anhydrous sodium sulfate drying.After drying, filter, filtrate concentrates, and silica gel column chromatography is separated, and with ethyl acetate: sherwood oil (60 ~ 90 DEG C)=1:4 (V:V) wash-out, obtains white fluffy solid 100mg, is compound g 1, yield 86.2%.
Step 2:3-O-(dimethyl tertiary butyl is silica-based)-21-O-demethyl Brusatol (E)
Get C and be about 18g(0.028mol), lithium iodide 18g(0.13mol) dissolve with 35ml anhydrous pyridine, under nitrogen protection, stirring reaction 13h at 100 DEG C.After reaction terminates, underpressure distillation removes most of pyridine, and enriched material is with 200ml diluted ethyl acetate.Organic layer washs with dilute hydrochloric acid, and pickle solution, with ethyl acetate (1 × 100ml) extraction, merges all organic phases, with anhydrous sodium sulfate drying.After drying, filter, filtrate concentrates, and obtains orange red crude product and is about 17.5g.Get crude product and be about 500mg through preparative HPLC (YMC ODS post, acetonitrile: containing 0.03%TFA water=60:40) separation, obtaining sterling and be about 325mg, is compd E, yield 65.5%.Step 3:21-O-[3-(2-oxygen-3-benzenesulfonyl-1,2,5-oxadiazole-4-oxygen)-propoxy-]-21-O-demethyl Brusatol (30)
Get E45mg(0.073mmol), g 150mg(0.12mmol), dissolve with 5mlDMF, then add K 2cO 350mg(0.36mmol), under argon shield, with 40 DEG C of stirring reaction 12h.After reaction terminates, underpressure distillation is except desolventizing, and change and dissolve with methylene dichloride, with dilute hydrochloric acid, saturated common salt solution washing, organic layer is with anhydrous sodium sulfate drying.After drying, filter, filtrate concentrates, and with tetrahydrofuran (THF) dilution, adds a small amount of Glacial acetic acid, adds the tetrahydrofuran solution of 1mol/LTBAF until desiliconization ether protecting group is complete in batches.Reaction stops, and reaction solution adds 30ml methylene dichloride, more respectively with rare HCl(1 × 30ml), saturated NaHCO 3solution (1 × 30ml), saturated aqueous common salt (1 × 30ml) washs, and organic layer is with anhydrous sodium sulfate drying.After drying, filter, filtrate concentrates, and be separated with Preparative TLC silica-gel plate, methylene dichloride: methyl alcohol=30:1 is developping agent, obtains white powdery solids and be about 10mg, is compound 30, two step total recoverys 17.5%.
The physical and chemical parameter of compound 30 is as follows:
1H NMR(400MHz,CDCl 3)δ8.05(2H,br d,J=7.6Hz,Ar-H),7.77(1H,t,J=7.6Hz,Ar-H),7.63(2H,t,J=7.6Hz,Ar-H),6.32(1H,d,J=11.6Hz H-15),6.08(1H,s,OH-3),5.64(1H,s,H-2’),4.80(1H,s,H-7),4.71(1H,d,J=7.6Hz,H a-20),4.55（2H,t,J=6.0Hz,OCH 2）,4.49,4.33(each1H,m,OCH 2),4.25(1H,d,J=2.8Hz,H-11),4.22(1H,s,H-12),3.78(1H,d,J=7.6Hz,H b-20),3.14(1H,d,J=11.6Hz,H-14),2.98(1H,d,J=16.0Hz,H β-1),2.95(1H,overlap,H-5),2.39(1H,d,J=16.0Hz,H α-1),2.39(1H,overlap,H α-6),2.30(2H,m,CH 2),2.19(3H,s,H-5’),2.12(1H,br s,H-9),1.93(3H,s,H-4’),1.84(3H,d,J=1.2Hz,H-18),1.76(1H,ddd,J=14.0,12.8,2.0Hz,H β-6),1.40(3H,s,H-19);
13C NMR(100MHz,CDCl 3)δ191.9(C-2),171.7(C-21),167.0(C-16),164.4(C-1’),161.4(C-3’),158.8(C=N),144.0(C-3),137.7(ArC),135.8(ArC),129.7(ArC×2)，128.6(ArC×2)，127.6(C-4)，113.9(C-2’),110.5(C=N),82.3(C-7),81.3(C-13),75.8(C-12),74.1(C-20),71.0(C-11),68.0(OCH 2),65.7(C-15),62.6(OCH 2),51.7(C-14),48.6(C-1),45.5(C-8),42.0(C-9),41.8(C-5),41.1(C-10),29.1(C-6),27.7(CH 2),27.7(C-4’),20.7(C-5’),15.4(C-19),13.3(C-18);
HRESIMS m/z811.1979[M+Na] +(calcd for C 36H 40N 2NaO 16S,811.1991).
Embodiment 31:
Synthetic route:
Step 1:21-O-demethyl Brusatol (F)
The E crude product 17g of gained in embodiment 30 and 100ml tetrahydrofuran (THF) are placed in reaction flask, and add the tetrahydrofuran solution of 40ml1mol/LTBAF, stirring is spent the night in batches.Reaction solution concentrates, and presses preparative column to be separated, with methyl alcohol: containing 0.03%TFA water=10:90 ~ 15:85 ~ 20:80 ~ 25:75 (V:V) gradient elution, to obtain white powdery solids and be about 7.1g in anti-phase ODS post.With the yield of Compound C in embodiment 32 for raw material calculating compound F 17-hydroxy-corticosterone, be compound F 17-hydroxy-corticosterone, two step total recoverys are 49.5%.
Step 2:21-O-[4-(2-oxygen-3-benzenesulfonyl-1,2,5-oxadiazole-4-oxygen)-butoxy]-21-O-demethyl Brusatol (31)
Get F24mg(0.047mmol), b 230mg(0.096mmol), EDCI18mg(0.094mmol), DMAP40mg(0.33mmol) and, in ice-water bath, add anhydrous methylene chloride and dissolve, after stirring 5h, change with stirred overnight at room temperature.Reaction stops, and reaction solution is with water, and rare HCl, saturated common salt water washing, organic layer is with anhydrous sodium sulfate drying.After drying, filter, organic layer concentrates, and be separated with preparative HPLC (YMC ODS post, acetonitrile: containing 0.03%TFA water=60:40), obtaining white powdery solids, about 9mg, is compound 31, yield 23.7%.
The physical and chemical parameter of compound 31 is as follows:
1H NMR(400MHz,CDCl 3)δ8.05(2H,br d,J=7.6Hz,Ar-H),7.77(1H,t,J=7.6Hz,Ar-H),7.63(2H,t,J=7.6Hz,Ar-H),6.28(1H,d,J=11.6Hz H-15),6.08(1H,s,OH-3),5.62(1H,s,H-2’),4.80(1H,s,H-7),4.69(1H,d,J=8.0Hz,H a-20),4.49(2H,t,J=4.8Hz,OCH 2),4.36,4.24(each1H,m,OCH 2),4.24(1H,br s,H-11),4.22(1H,s,H-12),3.78(1H,d,J=8.0Hz,H b-20),3.14(1H,d,J=11.6Hz,H-14),2.98(1H,d,J=16.0Hz,H β-1),2.94(1H,overlap,H-5),2.38(1H,d,J=16.0Hz,H α-1),2.39(1H,d,J=14.4,H α-6),2.19(3H,s,H-5’),2.11(1H,br s,H-9),2.00(2H,m,CH 2),1.93(2H,m,CH 2),1.92(3H,s,H-4’),1.84(3H,s,H-18),1.76(1H,ddd,J=14.4,12.8,2.0Hz,H β-6),1.39(3H,s,H-19);
13C NMR(100MHz,CDCl 3)δ192.0(C-2),171.6(C-21),167.0(C-16),164.4(C-1’),161.1(C-3’),159.0(C=N),144.0(C-3),137.8(ArC),135.7(ArC),129.7(ArC×2)，128.6(ArC×2)，127.7(C-4)，114.0(C-2’),110.6(C=N),82.3(C-7),81.2(C-13),75.8(C-12),74.0(C-20),71.0(C-11),70.8(OCH 2),65.8(C-15),65.7(OCH 2),51.6(C-14),48.6(C-1),45.5(C-8),41.9(C-9),41.8(C-5),41.1(C-10),29.1(C-6),27.7(C-4’),25.4(CH 2),24.8(CH 2),20.7(C-5’),15.4(C-19),13.3(C-18);
HRESIMS m/z803.2337[M+H] +(calcd for C 37H 43N 2O 16S,803.2328).
The synthetic route of compound 32,33 in embodiment: (intermediate a 7, a 8middle R 6, intermediate h 1, h 2middle R 6respectively with R in compound 32,33 9unanimously)
The end product (compound numbers corresponds to the compound numbers in embodiment) of synthesis:
Compound R 9
32 CH 2-p-Ph
33 CH 2-m-Ph
Embodiment 32:
Step 1:4-(3-benzenesulfonyl-1,2,5 oxadiazole-2-oxide compound-4-) oxygen benzyl bromine (h 1)
Get a 7330mg(0.95mmol) be placed in reaction flask, dissolve with 20ml anhydrous methylene chloride.Under ice-water bath condition, add excessive PBr 3, continue to react in ice-water bath.After 3h, change with stirred overnight at room temperature.After reaction terminates, in reaction solution, slowly add 20ml water, stir 10min.Get organic layer, wash with water (2 × 20ml).Water layer is with dichloromethane extraction 1 time.Merge all organic layers, with saturated NaHCO 3solution washs again to neutrality, and organic layer is with anhydrous sodium sulfate drying.After drying, filter, filtrate concentrates, and obtaining white solid 356mg, is compound h 1, productive rate 91.8%.
Step 2:21-O-[4-(2-oxygen-3-benzenesulfonyl-1,2,5-oxadiazole-4-oxygen)-benzyloxy]-21-O-demethyl Brusatol (32)
Get E70mg(0.11mmol), h 170mg(0.17mmol), K 2cO 3135mg(0.98mol), dissolve with 5ml anhydrous acetonitrile, under argon shield, 50 DEG C of stirring reaction 5h.After reaction stops, reacting liquid filtering, filtrate concentrates, and changes and dissolves, with rare HCl(1 × 20ml with 20ml methylene dichloride), saturated aqueous common salt (1 × 20ml) washs, and organic layer is with anhydrous sodium sulfate drying.After drying; filter, organic layer concentrates, and obtains 3-O-(dimethyl tertiary butyl silica-based with silica gel column chromatography separation)-21-O-[4-(2-oxygen-3-benzenesulfonyl-1; 2,5-oxadiazole-4-oxygen)-benzyloxy]-21-O-demethyl Brusatol crude product.Gained crude product dissolves with methylene dichloride 5ml, then adds 5ml tetrahydrofuran (THF) and a small amount of Glacial acetic acid, adds the tetrahydrofuran solution of 1mol/LTBAF until desiliconization ether protecting group is complete in batches.Reaction stops, and reaction solution adds 20ml methylene dichloride, more respectively with rare HCl(1 × 20ml), saturated NaHCO 3(1 × 20ml) solution, saturated aqueous common salt (1 × 20ml) washs, and organic layer is with anhydrous sodium sulfate drying.After drying, filter, organic layer concentrates.Be separated with preparative HPLC (YMC ODS post, acetonitrile: water=56:44), obtaining white powdery solids 50mg, is compound 32, two step total recoverys 53.0%.
The physical and chemical parameter of compound 32 is as follows:
1H NMR(400MHz,CDCl 3)δ8.10(2H,br d,J=8.0Hz,Ar-H),7.80(1H,t,J=8.0Hz,Ar-H),7.66(2H,t,J=8.0Hz,Ar-H),7.43(2H,d,J=8.4Hz,Ar-H),7.34(2H,d,J=8.4Hz,Ar-H),6.29(1H,d,J=11.6Hz,H-15),6.09(1H,s,OH-3),5.52(1H,s,H-2’),5.23,5.16(each1H,d,J=12.8Hz,OCH 2),4.80(1H,s,H-7),4.71(1H,d,J=7.6Hz,H a-20),4.25(1H,d,J=3.6Hz,H-11),4.23(1H,s,H-12),3.79(1H,d,J=7.6Hz,H b-20),3.17(1H,d,J=10.4Hz,H-14),2.96(1H,d,J=16.0Hz,H β-1),2.95(1H,overlap,H-5),2.38(1H,d,J=16.0Hz,H α-1),2.38(1H,overlap,H α-6),2.15(3H,s,H-5’),2.15(1H,overlap,H-9),1.89(3H,s,H-4’),1.83(3H,s,H-18),1.75(1H,ddd,J=14.4,12.8,2.4Hz,H β-6),1.38(3H,s,H-19);
13C NMR(100MHz,CDCl 3)δ192.1(C-2),171.3(C-21),167.0(C-16),164.4(C-1’),161.2(C-3’),158.2(C=N),152.6(ArC),144.0(C-3),137.8(ArC),135.9(ArC),133.3(ArC),129.8(ArC×2),129.8(ArC×2),128.6(ArC×2),127.9(C-4)，120.2(ArC×2)，113.9(C-2’),110.7(C=N),82.3(C-7),81.3(C-13),75.8(C-12),74.0(C-20),71.0(C-11),66.8(OCH 2)65.8(C-15),51.5(C-14),48.5(C-1),45.5(C-8),41.8(C-9),41.8(C-5),41.1(C-10),29.0(C-6),27.7(C-4’),20.6(C-5’),15.4(C-19),13.3(C-18);
HRESIMS m/z859.2008[M+Na] +(calcd for C 40H 40N 2NaO 16S,859.1991).
Embodiment 33:
21-O-[3-(2-oxygen-3-benzenesulfonyl-1,2,5-oxadiazole-4-oxygen)-benzyloxy]-21-O-demethyl Brusatol (33)
Step 1: with reference to step 1 in embodiment 32, with a 8replace a 7with PBr 3be obtained by reacting 3-(3-benzenesulfonyl-1,2,5 oxadiazole-2-oxide compound-4-) oxygen benzyl bromine (h 2).
Step 2: with reference to step 2 in embodiment 32, with h 2replace h 1, at K 2cO 3effect under, react with E, obtain 3-O-(dimethyl tertiary butyl by silica gel column chromatography silica-based)-21-O-[4-(2-oxygen-3-benzenesulfonyl-1,2,5-oxadiazole-4-oxygen)-benzyloxy]-21-O-demethyl Brusatol crude product.Gained crude product sloughs silicon ether protecting group with TBAF again, and be separated through preparative HPLC (YMC ODS post, acetonitrile: water=56:44), obtaining white powdery solids, is compound 33, two step total recoverys 30.9%.
The physical and chemical parameter of compound 33 is as follows:
1H NMR(400MHz,CDCl 3)δ8.10(2H,dd,J=8.0,1.2Hz,Ar-H),7.80(1H,t,J=8.0Hz,Ar-H),7.66(2H,t,J=8.0Hz,Ar-H),7.48(1H,t,J=8.0Hz,Ar-H),7.39(1H,br s,Ar-H),7.30(1H,dd,J=8.0,2.0Hz,Ar-H),7.29(1H,d,J=8.0,Ar-H),6.31(1H,d,J=11.6Hz,H-15),6.07(1H,s,OH-3),5.50(1H,s,H-2’),5.25,5.20(each1H,d,J=12.8Hz,OCH 2),4.80(1H,s,H-7),4.72(1H,d,J=8.0Hz,H a-20),4.26(1H,br s,H-11),4.23(1H,s,H-12),3.81(1H,d,J=8.0Hz,H b-20),3.17(1H,d,J=11.6Hz,H-14),2.98(1H,d,J=16.0Hz,H β-1),2.95(1H,d,J=12.0Hz,H-5),2.39(1H,d,J=16.0Hz,H α-1),2.39(1H,overlap,H α-6),2.15(3H,s,H-5’),2.12(1H,overlap,H-9),1.89(3H,s,H-4’),1.85(3H,d,J=1.6Hz,H-18),1.73(1H,ddd,J=13.6,12.0,2.4Hz,H β-6),1.36(3H,s,H-19);
13C NMR(100MHz,CDCl 3)δ192.0(C-2),171.2(C-21),167.0(C-16),164.4(C-1’),161.2(C-3’),158.1(C=N),152.9(ArC),144.0(C-3),137.6(ArC),137.1(ArC),135.9(ArC),130.4(ArC),129.8(ArC×2),128.7(ArC×2),127.8(C-4),125.8(ArC),119.8(ArC),119.0(ArC),113.9(C-2’),110.7(C=N),82.2(C-7),81.4(C-13),75.8(C-12),74.0(C-20),71.0(C-11),66.5(OCH 2),65.7(C-15),51.6(C-14),48.6(C-1),45.5(C-8),41.8(C-9),41.8(C-5),41.1(C-10),29.0(C-6),27.7(C-4’),20.6(C-5’),15.4(C-19),13.3(C-18);
HRESIMS m/z837.2204[M+H] +(calcd for C 40H 41N 2O 16S,837.2212).
The synthetic route of compound 34-36 in embodiment: (intermediate c 1-c 3middle R 7, intermediate e 1-e 3middle R 7respectively with R in compound 34-36 10unanimously)
The compound of synthesis:
Compound R 10
34 CH 2-p-Ph
35 CH 2-m-Ph
36 CH 2-o-Ph
Embodiment 34:
21-O-[4-(2-oxygen-4-phenyl-1,2,5-oxadiazole-3 methoxyl group)-benzyloxy]-21-O-demethyl Brusatol (34)
With reference to step 2 in embodiment 32, with gained e in step 1 in embodiment 15 1replace h 1, at K 2cO 3effect under, react with E, obtain 3-O-(dimethyl tertiary butyl is silica-based)-21-O-[4-(2-oxygen-4-phenyl-1,2,5-oxadiazole-3 methoxyl group)-benzyloxy]-21-O-demethyl Brusatol crude product by silica gel column chromatography.Gained crude product sloughs silicon ether protecting group with TBAF again, and be separated through preparative HPLC (YMC ODS post, acetonitrile: water=56:44), obtaining white powdery solids, is compound 34, two step total recoverys 60.9%.
The physical and chemical parameter of compound 34 is as follows:
1H NMR(400MHz,CDCl 3)δ7.86(2H,dd,J=8.0,1.2Hz,Ar-H),7.56(3H,m,Ar-H),7.32(2H,d,J=8.4Hz,Ar-H),7.02(2H,d,J=8.4Hz,Ar-H),6.32(1H,d,J=11.6Hz,H-15),6.07(1H,s,OH-3),5.54(1H,s,H-2’),5.16,5.11(each1H,d,J=12.4Hz,OCH 2),5.13(2H,s，OCH 2),4.78(1H,s,H-7),4.70(1H,d,J=8.0Hz,H a-20),4.25(1H,d,J=3.2Hz,H-11),4.20(1H,s,H-12),3.79(1H,d,J=8.0Hz,H b-20),3.10(1H,d,J=12.8Hz,H-14),2.98(1H,d,J=16.0Hz,H β-1),2.96(1H,d,J=12.8Hz,H-5),2.39(1H,d,J=16.0Hz,H α-1),2.39(1H,overlap,H α-6),2.17(3H,s,H-5’),2.13(1H,br s,H-9),1.91(3H,s,H-4’),1.85(3H,d,J=1.2Hz,H-18),1.76(1H,ddd,J=14.0,12.8,1.2Hz,H β-6),1.40(3H,s,H-19);
13C NMR(100MHz,CDCl 3)δ192.1(C-2),171.4(C-21),167.1(C-16),164.4(C-1’),161.1(C-3’),157.2(ArC),157.0(C=N),144.0(C-3),131.5(ArC),130.2(ArC×2),129.4(ArC×2),128.5(ArC),128.0(C-4),127.6(ArC×2),126.0(ArC),115.1(ArC×2),114.0(C-2’),112.0(C=N),82.3(C-7),81.3(C-13),75.8(C-12),73.9(C-20),71.0(C-11),67.4(OCH 2),65.7(C-15),58.4(OCH 2),51.5(C-14),48.5(C-1),45.5(C-8),41.8(C-9),41.8(C-5),41.1(C-10),29.0(C-6),27.7(C-4’),20.6(C-5’),15.4(C-19),13.3(C-18);
HRESIMS m/z787.2714[M+H] +(calcd for C 41H 43N 2O 14,787.2709).
Embodiment 35:
21-O-[3-(2-oxygen-4-phenyl-1,2,5-oxadiazole-3 methoxyl group)-benzyloxy]-21-O-demethyl Brusatol (35)
Step 1: with reference to step 1 in embodiment 15, with c 2replace c 1, with PBr 3be obtained by reacting 3-(4-phenyl-1,2,5-oxadiazole-2-oxide compound-3) methoxybenzyl bromine (e 2).
Step 2: with reference to step 2 in embodiment 32, with e 2replace h 1, at K 2cO 3effect under, react with E, obtain 3-O-(dimethyl tertiary butyl by silica gel column chromatography silica-based)-21-O-[3-(2-oxygen-4-phenyl-1,2,5-oxadiazole-3 methoxyl group)-benzyloxy]-21-O-demethyl Brusatol crude product.Gained crude product sloughs silicon ether protecting group with TBAF again, and be separated through preparative HPLC (YMC ODS post, acetonitrile: water=56:44), obtaining white powdery solids, is compound 35, two step total recoverys 60.3%.The physical and chemical parameter of compound 35 is as follows:
1H NMR(400MHz,CDCl 3)δ7.90(2H,dd,J=8.0,1.2Hz,Ar-H),7.57(3H,m,Ar-H),7.31(1H,t,J=8.4Hz,Ar-H),7.14(1H,br s,Ar-H),6.98(2H,m,Ar-H),6.32(1H,brs,H-15),6.08(1H,s,OH-3),5.30,5.14(each1H,d,J=12.8Hz,OCH 2),5.25(1H,s,H-2’),5.22,5.14(each1H,d,J=13.2Hz,OCH 2),5.13(2H,s,OCH 2),4.91(1H,s,H-7),4.73(1H,d,J=7.6Hz,H a-20),4.26(1H,br s,H-11),4.24(1H,s,H-12),3.94(1H,d,J=7.6Hz,H b-20),3.39(1H,d,J=13.2Hz,H-14),3.00(1H,d,J=16.4Hz,H β-1),2.96(1H,d,J=12.0Hz,H-5),2.40(1H,d,J=16.4Hz,H α-1),2.39(1H,overlap,H α-6),2.18(1H,brs,H-9),2.11(3H,s,H-5’),1.85(3H,d,J=1.2Hz,H-18),1.80(3H,s,H-4’),1.76(1H,overlap,H β-6),1.42(3H,s,H-19);
13C NMR(100MHz,CDCl 3)δ192.2(C-2),171.4(C-21),167.5(C-16),164.3(C-1’),161.0(C-3’),157.2(ArC),157.1(C=N),144.0(C-3),137.0(ArC),131.5(ArC),130.1(ArC),129.4(ArC×2),128.0(C-4),127.7(ArC×2),126.0(ArC),121.4(ArC),116.1(ArC),113.8(C-2’),113.2(ArC),112.6(C=N),82.3(C-7),81.4(C-13),75.7(C-12),74.0(C-20),71.1(C-11),67.0(OCH 2),65.8(C-15),58.4(OCH 2),51.2(C-14),48.5(C-1),45.6(C-8),41.9(C-9),41.9(C-5),41.1(C-10),29.0(C-6),27.6(C-4’),20.6(C-5’),15.4(C-19),13.3(C-18);
HRESIMS m/z787.2720[M+H] +(calcd for C 41H 43N 2O 14,787.2709).
Embodiment 36:
21-O-[2-(2-oxygen-4-phenyl-1,2,5-oxadiazole-3 methoxyl group)-benzyloxy]-21-O-demethyl Brusatol (36)
Step 1: with reference to step 1 in embodiment 15, with c 3replace c 1with PBr 3be obtained by reacting 3-(4-phenyl-1,2,5-oxadiazole-2-oxide compound-3) methoxybenzyl bromine (e 3).
Step 2: with reference to step 2 in embodiment 32, with e 3replace h 1, at K 2cO 3effect under, react with E, obtain 3-O-(dimethyl tertiary butyl by silica gel column chromatography silica-based)-21-O-[2-(2-oxygen-4-phenyl-1,2,5-oxadiazole-3 methoxyl group)-benzyloxy]-21-O-demethyl Brusatol crude product.Gained crude product sloughs silicon ether protecting group with TBAF again, and be separated through preparative HPLC (YMC ODS post, acetonitrile: water=56:44), obtaining white powdery solids, is compound 36, two step total recoverys 53.3%.
The physical and chemical parameter of compound 36 is as follows:
1H NMR(400MHz,CDCl 3)δ7.80(2H,br d,J=7.6Hz,Ar-H),7.55(3H,m,Ar-H),7.38(2H,m,Ar-H),7.06(2H,m,Ar-H),6.31(1H,br s,H-15),6.07(1H,s,OH-3),5.33(1H,s,H-2’),5.21,5.17(each1H,d,J=12.0Hz,OCH 2),5.13(2H,s,OCH 2),4.76(1H,s,H-7),4.62(1H,d,J=8.0Hz,H a-20),4.19(1H,br s,H-11),4.10(1H,s,H-12),3.71(1H,d,J=8.0Hz,H b-20),3.08(1H,d,J=11.6Hz,H-14),2.94(1H,d,J=16.0Hz,H β-1),2.92(1H,overlap,H-5),2.36(1H,overlap,H α-6),2.33(1H,d,J=16.0Hz,H α-1),2.11(3H,s,H-5’),2.09(1H,br s,H-9),1.83(3H,s,H-4’),1.79(3H,s,H-18),1.76(1H,ddd,J=13.6,12.8,1.2Hz,Hb-6),1.37(3H,s,H-19);
13C NMR(100MHz,CDCl 3)δ192.0(C-2),171.5(C-21),167.0(C-16),164.3(C-1’),160.7(C-3’),156.9(C=N),155.5(ArC),144.0(C-3),131.6(ArC),130.9(ArC),130.6(ArC),129.5(ArC×2),127.7(C-4),127.6(ArC×2),126.0(ArC),123.7(ArC),122.4(ArC),113.9(C-2’),112.1(ArC),111.9(C=N),82.0(C-7),81.3(C-13),75.8(C-12),73.9(C-20),70.9(C-11),65.5(C-15),63.4(C-1’’),58.6(OCH 2),51.4(C-14),48.6(C-1),45.4(C-8),41.9(C-9),41.8(C-5),41.1(C-10),29.0(C-6),27.5(C-4’),20.6(C-5’),15.4(C-19),13.3(C-18);
HRESIMS m/z787.2707[M+H] +(calcd for C 41H 43N 2O 14,787.2709).
Embodiment 37:
Synthetic route:
21-O-[(4-phenyl-1,2,5-oxadiazole-2-oxide compound-3) methoxyl group]-21-O-demethyl Brusatol (37)
Step 1: with reference to step 1 in embodiment 15, replaces c with B 1with PBr 3be obtained by reacting 2-oxygen-3-brooethyl-4-phenyl-1,2,5-oxadiazole (e 4).
Step 2: with reference to step 2 in embodiment 32, with e 4replace h 1, then at K 2cO 3effect under, react with E, obtain 3-O-(dimethyl tertiary butyl by silica gel column chromatography silica-based)-21-O-[(4-phenyl-1,2,5-oxadiazole-2-oxide compound-3) methoxyl group]-21-O-demethyl Brusatol crude product.Gained crude product sloughs silicon ether protecting group with TBAF again, and be separated through preparative HPLC (YMC ODS post, acetonitrile: water=52:48), obtaining white powdery solids, is compound 37, two step total recoverys 48.0%.
The physical and chemical parameter of compound 37 is as follows:
1H NMR(400MHz,CDCl 3)δ7.75(2H,dd,J=7.6,1.2Hz,Ar-H),7.57(3H,m,Ar-H),6.26(1H,d,J=11.6Hz H-15),6.07(1H,s,OH-3),5.46(1H,s,H-2’),5.31,5.23(each1H,d,J=13.6Hz,OCH 2),4.66(1H,d,J=8.0Hz,H a-20),4.62(1H,s,H-7),4.23(1H,d,J=3.2Hz,H-11),4.17(1H,s,H-12),3.59(1H,d,J=8.0Hz,H b-20),3.23(1H,br s,H-14),2.94(1H,d,J=16.0Hz,H β-1),2.92(1H,d,J=12.0Hz,H-5),2.38(1H,d,J=16.0Hz,H α-1),2.36(1H,d,J=14.8Hz,H α-6),2.15(3H,s,H-5’),2.12(1H,br s,H-9),1.88(3H,s,H-4’),1.83(3H,s,H-18),1.76(1H,ddd,J=14.0,12.0,1.2Hz,H β-6),1.36(3H,s,H-19);
13C NMR(100MHz,CDCl 3)δ192.0(C-2),170.7(C-21),166.9(C-16),164.3(C-1’),162.0(C-3’),156.5(C=N),144.0(C-3),131.5(ArC),129.5(ArC×2),127.9(C-4),127.8(ArC×2),125.9(ArC),113.4(C-2’),110.6(C=N),82.1(C-7),81.4(C-13),75.6(C-12),74.1(C-20),71.1(C-11),65.4(C-15),55.7(OCH 2),51.6(C-14),48.5(C-1),45.4(C-8),41.8(C-9),41.8(C-5),41.0(C-10),29.0(C-6),27.6(C-4’),20.7(C-5’),15.4(C-19),13.3(C-18);
HRESIMS m/z703.2108[M+Na] +(calcd for C 34H 36N 2NaO 13,703.2110).
The synthetic route of compound 38-40 in embodiment:
Change F and b 2feed ratio, 38 of different ratios can be obtained, 39,40
The end product (compound numbers corresponds to the compound numbers in embodiment) of synthesis:
Embodiment 38:
12-O-{4-[4-(2-oxygen-3-benzenesulfonyl-1,2,5-oxadiazole-4-oxygen)-butoxy]-4-oxobutanoyl }-21-O-demethyl Brusatol (38)
By F28mg (0.055mmol), b 235mg (0.085mmol), EDCI20mg(0.10mmol), DMAP50mg(0.41mmol) be placed in reaction flask, under ice-water bath condition, add anhydrous methylene chloride and dissolve, after stirring reaction 5h, change and spend the night with stirring at room temperature reaction.Reaction solution respectively with rare HCl, rare NaHCO 3solution, saturated common salt water washing.Rare NaHCO 3washing lotion adjusts pH to 4 with rare HCl, then with dichloromethane extraction 1 time.Merge all organic layers, with anhydrous sodium sulfate drying.After drying, filter, filtrate concentrates, and be separated through preparative HPLC (YMC ODS post, acetonitrile: containing 0.03%TFA water=50:50), obtaining white powdery solids, about 17mg, is compound 38, yield 34.1%.(this embodiment also can obtain minority specioz 43,44)
The physical and chemical parameter of compound 38 is as follows:
1H NMR(500MHz,CDCl 3)δ8.05(2H,br d,J=8.0Hz,Ar-H),7.77(1H,t,J=8.0Hz,Ar-H),7.63(2H,t,J=8.0Hz,Ar-H),6.08(1H,d,J=11.6Hz,H-15),6.08(1H,overlap,OH-3),5.68(1H,s,H-2’),5.26(1H,s,H-12),4.82(1H,s,H-7),4.78(1H,d,J=6.5Hz,H a-20),4.45(2H,t,J=6.0Hz,OCH 2),4.19(1H,br s,H-11),4.15(2H,m,OCH 2),3.80(1H,d,J=6.0Hz,H b-20),3.25(1H,br s,H-14),2.97(1H,d,J=13.0Hz,H-5),2.92(1H,d,J=17.0Hz,H β-1),2.67(1H,d,J=17.0Hz,H α-1),2.62(1H,overlap,H α-6),2.52(4H,m,COCH 2CH 2CO),2.16(3H,s,H-5’),2.16(1H,overlap,H-9),1.95(2H,m,CH 2),1.90(3H,s,H-4’),1.84(3H,s,H-18),1.81(2H,m,CH 2),1.78(1H,overlap,H β-6),1.37(3H,s,H-19);
13C NMR(125MHz,CDCl 3)δ192.3(C-2),172.4(C=O),171.5(C-21),170.9(C=O),167.1(C-16),164.7(C-1’),160.8(C-3’),158.9(C=N),144.0(C-3),137.9(ArC),135.7(ArC),129.7(ArC×2)，128.6(ArC×2)，128.4(C-4)，114.2(C-2’),110.5(C=N),82.6(C-7),80.1(C-13),74.7(C-12),74.0(C-20),71.0(OCH 2),68.9(C-11),65.5(C-15),64.3(OCH 2),51.3(C-14),48.4(C-1),45.2(C-8),42.0(C-9),41.8(C-5),41.0(C-10),29.3(C-6),29.0(CO CH 2),28.8(CO CH 2),27.6(C-4’),25.2(CH 2),24.9(CH 2),20.5(C-5’),15.5(C-19),13.4(C-18);
HRESIMS m/z925.2316[M+Na] +(calcd for C 41H 46N 2NaO 19S,925.2308).
Embodiment 39:
3,12-O-bis-{ 4-[4-(2-oxygen-3-benzenesulfonyl-1,2,5-oxadiazole-4-oxygen)-butoxy]-4-oxobutanoyl }-21-O-demethyl Brusatol (39)
By b 26.3g (15.2mmol), EDCI2.96g(15.4mmol), DMAP7g(57.3mmol) reaction flask is placed in, under ice-water bath condition, add anhydrous methylene chloride to dissolve, after stirring 3h, add F3.85g(7.61mmol), continue stirring reaction 2h under ice-water bath, then change with room temperature reaction.After 12h, add b 21.1g (2.7mmol), EDCI0.55g(2.87mmol), DMAP0.5g(4.1mmol), continue stirring at room temperature reaction 24h.After reaction terminates, reaction solution concentrates, and Sephadex LH-20 gel column chromatography is separated, with methylene dichloride: methyl alcohol=1:1(V:V) wash-out obtains crude product 5.2g.Crude product for ODS pillar layer separation, with methyl alcohol: water=65:35 (V:V) wash-out, obtains white powdery solids 3.3g, is compound 39 with middle compacting, yield 33.4%.The physical and chemical parameter of compound 39 is as follows:
1H NMR(500MHz,CDCl 3)δ8.04(4H,br d,J=7.5Hz,Ar-H),7.75(2H,t,J=7.5Hz,Ar-H),7.63(4H,t,J=7.5Hz,Ar-H),6.08(1H,br s,H-15),5.67(1H,s,H-2’),5.25(1H,s,H-12),4.83(1H,s,H-7),4.76(1H,d,J=7.5Hz,H a-20),4.45(4H,m,OCH 2×2),4.19(1H,overlap,H-11),4.19(2H,t,J=6.5Hz,OCH 2),4.14(2H,m,OCH 2),3.79(1H,d,J=7.5Hz,H b-20),3.26(1H,br s,H-14),3.06(1H,d,J=13.0Hz,H-5),2.89(1H,d,J=17.5Hz,H β-1),2.87(2H,t,J=6.5Hz,COCH 2),2.71(2H,m,COCH 2),2.66(1H,overlap,H α-6),2.65(1H,overlap,H α-1),2.53(4H,m,COCH 2CH 2CO),2.15(3H,s,H-5’),2.15(1H,overlap,H-9),1.94(4H,m,CH 2×2),1.90(3H,s,H-4’),1.82(4H,m,CH 2×2),1.80(3H,s,H-18),1.78(1H,overlap,H β-6),1.44(3H,s,H-19);
13C NMR(125MHz,CDCl 3)δ189.3(C-2),172.5(C=O),172.0(C=O),171.4(C-21),170.5(C=O),170.3(C=O),167.1(C-16),164.7(C-1’),160.9(C-3’),158.9(C=N×2),146.2(C-3),142.0(C-4),138.0(ArC×2),135.7(ArC×2),129.7(ArC×4),128.5(ArC×4),114.2(C-2’),110.5(C=N×2),82.3(C-7),80.1(C-13),74.6(C-12),73.7(C-20),71.0(OCH 2×2),68.8(C-11),65.5(C-15),64.3(OCH 2),64.0(OCH 2),51.2(C-14),49.8(C-1),45.0(C-8),42.8(C-5),41.9(C-9),40.7(C-10),29.3(C-6),29.0(CO CH 2),28.8(CO CH 2×2),28.5(CO CH 2),27.6(C-4’),25.1(CH 2×2),24.9(CH 2),24.9(CH 2),20.6(C-5’),15.5(C-19),14.5(C-18);
HRESIMS m/z1297.2962[M-H] -(calcd for C57H61N4O27S2,1297.2970).
Embodiment 40:
3-O-{4-[4-(2-oxygen-3-benzenesulfonyl-1,2,5-oxadiazole-4-oxygen)-butoxy]-4-oxobutanoyl }-21-O-demethyl Brusatol (40)
By the operation of embodiment 39, when with Sephadex LH-20 gel chromatography separation 39, except obtaining 38, obtaining another white powdery solids and be about 520mg, is compound 40, yield 7.6%.(this embodiment also can obtain minority specioz 38)
The physical and chemical parameter of compound 40 is as follows:
1H NMR(500MHz,CDCl 3)δ8.04(2H,br d,J=7.5Hz，ArH),7.76(1H,t,J=7.5Hz,ArH),7.62(2H,t,J=7.5Hz,ArH),6.30(1H,br s,H-15),5.66(1H,s,H-2’),4.82(1H,s,H-7),4.69(1H,br s,H a-20),4.44(2H,t,J=6.0Hz，OCH 2),4.19(2H,t,J=6.0Hz，OCH 2),4.19(1H,overlap,H-11),4.11(1H,s,H-12),3.77(1H,br s,Hb-20),3.07(1H,d,J=10.5Hz,H-14),3.07(1H,overlap,H-5),2.91(1H,overlap,H β-1),2.88(2H,t,J=6.0Hz,COCH 2),2.71(2H,t,J=6.0Hz,COCH 2),2.49(1H,d,J=14.0Hz,H α-1),2.34(1H,d,J=12.0Hz,H α-6),2.19(1H,br s,H-9),2.12(3H,s,H-5’),1.94(2H,m,CH 2),1.88(3H,s,H-4’),1.82(2H,m,CH 2),1.80(1H,overlap,H β-6),1.80(3H,s,H-18),1.41(3H,s,H-19);
13CNMR(125MHz,CDCl 3)δ190.2(C-2),172.7(C-21),172.2(C=O),170.6(C=O),167.3(C-16),165.0(C-1’),161.0(C-3’),158.9(C=N),147.3(C-3),141.9(C-4),137.9(ArC),135.7(ArC),129.7(ArC×2),128.5(ArC×2),114.2(C-2’),110.5(C=N),82.3(C-7),80.9(C-13),75.7(C-12),73.7(C-20),71.0(C-11),70.7(OCH 2),65.8(C-15),64.2(OCH 2),51.2(C-14),49.7(C-1),45.3(C-8),42.8(C-5),41.2(C-9),40.7(C-10),29.0(C-6),28.7(CO CH 2),28.5(CO CH 2),27.6(C-4’),25.2(CH 2),24.9(CH 2),20.7(C-5’),15.5(C-19),14.6(C-18)；
HRESIMS m/z925.2266[M+Na] +(calcd for C 41H 46N 2NaO 19S,925.2308).
The anti-inflammatory of the compounds of this invention and the pharmacological experimental method of immunosuppressive activity and result following (compound numbers of pharmacological evaluation part corresponds to the compound numbers in embodiment):
Experimental example 1, compounds towards macrophages NO generate inhibit activities
Scavenger cell, performs body non-specific immune function, can produce the inflammatory factors such as NO under the inductions such as bacteria lipopolysaccharide LPS, participates in and inducing inflammatory reaction, in inflammation immunologic process initial stage and pathological development process, all has higher level.By detecting the mouse macrophage NO growing amount of original cuiture, can be used as external preliminary observation and screening has the component of certain anti-inflammatory activity or the indicator and model of compound.
Experimental technique:
1), experiment is divided into blank group 1 (only adding enchylema), blank group 2 (adding enchylema and sample), model group, administration group, positive control drug group.
2), C57BL6/J mouse peritoneal injection 4% sodium thioglycollate, by mouse sacrificed by decapitation after 4 days, drain blood.Abdominal injection PBS physiological buffer, liquid in sucking-off abdominal cavity, centrifugal abandoning supernatant, cell RPMI1640 is resuspended, and cell concn is adjusted to 1 × 10 by counting cells 6cellmL -1.
3), inoculating cell is in 48 porocyte culture plates.37 DEG C, 5%CO 2adherent culture makes cell attachment.Incline substratum, and rinse twice with PBS physiological buffer, remove non-attached cell, it is stand-by that every hole adds RPMI1640 substratum 500 μ l/ hole.
4), blank group only adds substratum, and model group adds LPS (final concentration is 1 μ g/ml), and administration group, except adding with except model group equivalent LPS, adds medicine to be measured in addition, and final concentration is 10 -5m, 37 DEG C of 5%CO 2continue cultivation 24 hours.
5), get 100 μ l supernatant liquors and equivalent Griess reagent mixes on micro oscillator, microplate reader measures 570nm reading, according to NO 2 -typical curve calculates NO 2 -content, later sample absorbance value is all converted into NO according to this typical curve 2 -content.
6), in aforesaid operations process, conventional mtt assay is also adopted to test positive control drug and medicine to be measured to the growth inhibition ratio of scavenger cell
Note: detect with blank group 2, finds that compound self can discharge the NO of trace, but the NO that after all stimulating well below model group LPS, scavenger cell produces, avoid the appearance of false negative result.
Experimental result:
Table 1
Note: F is 21-O-demethyl Brusatol
As can be seen from Table 1, compare with lead compound Brusatol, through the compound of structure of modification, or toxicity reduces or increased activity, and selectivity index is all higher than lead compound Brusatol.Especially compound 15,38,39,40, not only has and significantly generates inhibit activities to scavenger cell NO, also have the selectivity index apparently higher than lead compound Brusatol.
Experimental example 2, compound are to the immunosuppressive activity of mouse spleen lymphocyte propagation, conversion
Experimental technique:
1), experiment is divided into blank group (only adding enchylema), Con A group, LPS group, administration A group (only adding medicine to be measured), administration B group (adding Con A and medicine to be measured), administration C group (adding LPS and medicine to be measured).2), by after mice spleen taking-up pulverize, sieve, rinse 3 times with RPMI1640 nutrient solution.
3), erythrocyte cracked liquid splitting erythrocyte is used, the centrifugal 5min of 1000 turns/min.Centrifugal rear PBS is resuspended, counting cells.
4), inoculating cell in 96 porocyte culture plates, 1 × 10 6cells/well.
5), blank group, Con A group, LPS group, only add substratum 100 μ l.It is the RPMI1640 nutrient solution 100 μ l of 10 μMs of test-compounds that administration A, B, C group then adds containing concentration.After 1h, Con A group and administration B group add ConA (final concentration is 5 μ g/ml), transform for inducing T cell, LPS group and administration C group add LPS (final concentration is 10 μ g/ml), transform, put 5%CO for elicit B cell 2, cultivate 72h for 37 DEG C.
6), before termination is cultivated 4h, all add MTT (final concentration is 5mg/ml) to each group, after 4h, add 100 μ lDMSO, in microplate reader 570nm place reading OD value.Administration group, according to relevant control experiment, calculates splenocyte growth inhibition ratio respectively, B cell transforms inhibiting rate, T cell transforms inhibiting rate.
Experimental result:
Table 2
Note: Dex is positive control drug dexamethasone sodium phosphate
From result in table 2, during administration concentration 10 μMs, compound 10,15 pairs of splenocyte growth inhibition ratios are starkly lower than positive control drug (DEX) and Brusatol (Brusatol), show that toxicity is lower.Meanwhile, compound 10,15 all have higher B cell transforms inhibiting rate and T cell conversion inhibiting rate, and show that compound can suppress B cell, T cell transforms, and has obvious immunosuppressive activity.
Experimental example 3, part of compounds cause the impact of otitis activity to mouse knoting oil
Experimental technique:
Adopt mouse ear Oleum Tiglii to urge swelling model, observe test medicine and inhibit activities is regulated to acute inflammation.
Select male KM mouse, be divided into Normal group, model group, administration group at random, often organize 10, gastric infusion, successive administration 5 days.Administration group gives test-compound, and model group is only filled with distilled water.Subcutaneous administrations, dosage 10mg/kg, after 1 hour, 2% Oleum Tiglii 50ul is smeared on mouse right ear two sides, puts to death animal after 4 hours.Cut left and right auricle, take off auricle with the punch tool of diameter 8mm, weigh, with the difference of L-R auricle weight instruction ear thickness.Using gavage distilled water group as blank group, computerized compound is to the inhibiting rate of mice ear.
Experimental result:
Table 3
Compound number Dosage (mg/kg) Inhibiting rate (%)
Brusatol 10mg/kg is subcutaneous × and 1 37***
2 10mg/kg is subcutaneous × and 1 54***
8 10mg/kg is subcutaneous × and 1 53.2***
Note: compared with model group, * * p ﹤ 0.01; * * p ﹤ 0.001
From result in table 3, subcutaneous administration 1 time, during concentration 10mg/kg, compound 2, the inhibiting rate of 8 pairs of mice ear is apparently higher than primer Brusatol.
Experimental example 4, part of compounds cause the inhibit activities of mouse allergic dermatitis to DNF (DNFB).
Dinitrofluorobenzene is a kind of haptens, is combined into complete antigen makes sensitization of skin after being applied in skin of abdomen with skin protein, strengthens once after 24h, and sensitization is applied in ear on the 5th day again, causes delayed allergy.Experiment purpose is to observe compound to the restraining effect of mouse allergic dermatitis, its immunosuppressive activity of preliminary assessment.
Experimental technique:
Select ICR mouse, be divided into Normal group, model control group, administration group at random, often organize 10.Test-compound administration every day.Belly loses hair or feathers, 1%DNFB is evenly coated in belly sensitization, after 24h, same procedure is strengthened once, and sensitization the 5th day, attacks 1%DNFB uniform application in mouse right ear, mouse is put to death after 24h, cut left and right ear, make auricle with diameter 8mm blunderbuss, weigh, represent swelling with left and right auricle weight difference, calculate inhibitory rate of intumesce (%); Take mouse thymus and spleen, weigh, the ratio of internal organs weight in wet base and body weight represents organ index, calculates organ index inhibiting rate (%), for the impact of assessing compound on immunologic function.Organ index inhibiting rate is higher, shows that immunosuppression capability is stronger, but too high inhibiting rate also points out compound excessively strong to the toxicity of internal organs.
Experimental result:
Table 4.
note: compared with model group, * p ﹤ 0.05; * * p ﹤ 0.001
Result display in table 4, with 1mg/kg dosage, subcutaneous administrations, successive administration 6 days, compound 10 pairs of mice ear inhibit activities are obviously better than primer Brusatol, simultaneously to the inhibiting rate of thymus index and index and spleen index significantly lower than primer Brusatol, prompting systemic immunity inhibited reaction is less, to the toxicity of internal organs lower than Brusatol.
Table 5
Note: compared with model group, * p ﹤ 0.05; * p ﹤ 0.01; * * p ﹤ 0.001
Result display in table 5, with 2mg/kg dosage, oral administration gavage administration, successive administration 6 days, compound 2 pairs of mice ear inhibit activities are better than primer Brusatol, simultaneously to the inhibiting rate of thymus index significantly lower than primer Brusatol prompting to the toxicity of thymus gland lower than Brusatol, but index and spleen index inhibiting rate has no significant difference.
Table 6
Note: compared with model group, * p ﹤ 0.05; * p ﹤ 0.01; * * p ﹤ 0.001
Result display in table 6, with 2mg/kg dosage, oral administration gavage administration, successive administration 6 days, compound 34,39 pairs of mice ear inhibit activities are better than primer Brusatol, to the inhibiting rate of thymus index and index and spleen index significantly lower than primer Brusatol, prompting systemic immunity inhibited reaction is less, to the toxicity of internal organs lower than Brusatol.
Experimental example 5. part of compounds is to the inhibit activities of OVA inducing mouse inflammation of asthma
Experimental technique:
I. group and dosage are arranged:
Blank group: equal-volume solvent;
Model group: equal-volume solvent;
Dexamethasone sodium phosphate (516.41): 1 μm of ol/Kg (0.516mg/Kg);
Brusatol(520)：4μmol/Kg(2.08mg/Kg)；
Compound 2 (916): 4 μm of ol/Kg (3.664mg/Kg).
Compound 10 (900): 4 μm of ol/Kg (3.6mg/Kg);
Ii. the foundation of model:
Animal is divided into 7 groups at random, every 20, weighs; Except blank group gives isopyknic physiological saline, other each treated animal abdominal injection OVA solution 50 μ L(20 μ g) carry out sensitization with the mixed solution of aluminum hydroxide gel 50 μ L; After sensitization the 7th, 14 day first, except blank group gives isopyknic physiological saline, each treated animal respectively OVA of abdominal injection sensitization same dose and aluminum hydroxide gel carried out immune strengthening; After the 14th day immune strengthening 1 hour, animal arranged shown gastric infusion by group and dosage, once a day, and continuous 14 days; In the 26th day, anesthetized mice, the OVA solution 50 μ L of tracheal instillation 1.60mg/ml attacks, once a day, for three days on end; After last attacks 24h, put to death animal, draw materials and carry out Indexs measure.
Iii. Testing index and method:
Bronchoalveolar lavage fluid Arneth's count:
Animal is put to death with after Sodital intraperitoneal injection anesthesia, dissect neck, expose tracheae, with No. 4 half syringe needle promoting the circulation of qi cannulas, with 0.8ml precooling normal saline flushing 3 times, often all over rinsing 3 times back and forth, reclaim irrigating solution (rate of recovery reaches 80%), in 4 DEG C of centrifugal 10min of 1500rpm/min, collect supernatant,-20 DEG C frozen, for cytokines measurement; Contain the PBS re-suspended cell precipitation of 1%BSA with 0.5ml, on five classification differential blood count instrument, carry out Arneth's count.
In bronchoalveolar lavage fluid, the ELISA of relevant inflammatory factors measures:
Get-20 DEG C of frozen irrigating solutions, in thawing on ice, the ELISA kit specification sheets strictly with reference to the corresponding factor operates.
In serum, the ELISA of IgE level measures:
Animal socket of the eye venous blood collection, the centrifugal 15min of 3000rpm/min, separation of serum, is placed in-70 DEG C of cryogenic refrigerators and preserves, and during mensuration, strict reference ELISA kit specification sheets operates.
The mensuration of the propagation conversion capability of splenic lymphocyte:
After sacrifice, in 75% ethanol, soak 5min, in the aseptic taking-up spleen of super quiet worktable, shred, with pintle grinding, the serum-free medium of precooling rinses, and crosses unicellular mesh screen, prepares spleen single cell suspension, be adjusted to suitable cell density, be inoculated in 96 orifice plates, 5x10 5/ hole, often group establishes 4 multiple holes; Except blank group gives equal-volume solvent, each group adds LPS final concentration 10 μ g/ml or ConA final concentration 5 μ g/ml respectively, in 37 DEG C, and 5%CO 248h is cultivated under condition; Terminate front 4h in cultivation, every hole adds WST-8 solution 20ul, and after continuing to cultivate 4h, vibration mixing, measures absorbancy (A) in microplate reader 450nm place, calculates lymphocytic proliferation rate (%).
Lung tissue disease's Neo-Confucianism checks:
Zootomy chest cuts open, and gets middle lobe of right lung tissue, after 10% formaldehyde solution is fixing, carries out paraffin embedding, section, HE dyeing, makes histopathology and change inspection.
Experimental result
I. on the impact of the weight of animals change
As shown in Figure 1, compare with model group, after Brusatol and positive drug DEX administration, significantly suppress the weight of animals to increase, and other each compound affects unknown significance difference to body weight gain.
Ii. on the impact of asthmatic mouse pathological change
As shown in Fig. 2 result, compared with model group, the pathologic such as cellular infiltration, alveolar structure destruction that can significantly reduce inflammation after Brusatol, compound 2 and positive control drug DEX administration changes, and prompting has the effect of significant anti-asthma inflammation damnification; Compound 10 pairs of pathologics change then unknown significance and improve.Iii. on the impact that mice serum IgE secretes
As shown in Fig. 3 result, compared with model group, can significantly suppress the serum IgE level of mouse asthma to increase after compound 2 and positive control drug DEX administration, prompting has significant anti-asthma effect.
Iv. on the impact of mice spleen lymphocytic hyperplasia situation
As shown in Fig. 4 A result, compared with model group, the cellular immunization that can significantly suppress LPS to induce after compound 10 and positive control drug DEX administration; As shown in Fig. 4 B result, compared with model group, the humoral immunization that can significantly suppress ConA to induce after Brusatol, compound 2, compound 10 and DEX administration, prompting has the effect alleviating mouse asthma antibody tormation.
V. on the impact of bronchoalveolar lavage fluid Lymphocyte subset
As shown in Fig. 5 result, compared with model group, after Brusatol, compound 2 and positive control drug DEX administration, significantly can reduce the total white blood cells in irrigating solution and neutrophil leucocyte number; And compound 2 significantly can reduce the eosinophilic granulocyte in irrigating solution; Result prompting compound 2 can significantly alleviate mouse asthmatic inflammation.
Vi. on the impact of mouse asthma bronchoalveolar lavage fluid inflammatory factor secretion situation
As shown in Fig. 6 result, compared with model group, can the secretion of remarkable inflammation-inhibiting factor IL-1 β, IL-6 and TNF-α after Brusatol, compound 2 and positive control drug DEX administration, and Brusatol significantly promotes the secretion of anti-inflammatory factors IL-10, compound 2 has the trend promoting IL-10 secretion, but there was no significant difference, result prompting Brusatol and compound 2 have the effect of anti-asthma inflammation.
Experimental example 6. part of compounds is to the inhibit activities of murine chronic obstructive pneumonia
Experimental technique:
I. group and dosage are arranged:
Blank group: equal-volume solvent;
Model group: equal-volume solvent;
Dexamethasone sodium phosphate (516.41): 1 μm of ol/Kg;
Brusatol(520)：2μmol/kg；
Compound 2 (916): 2 μm of ol/kg.
Compound 39 (1298): 2 μm of ol/kg;
Ii. the foundation of model:
Animal is divided into 7 groups at random, often organizes 20, weighs; Except blank group gives isopyknic physiological saline, other each group gives as group and dosage arrange shown drug dose, gastric infusion, every day 1 time (dexamethasone administration time section be 26 ?35 days, give equal-volume physiological saline At All Other Times); In administration in the 1st, 14 day after 1 hour, after Animal Anesthesia, tracheae instills the normal saline solution that a 30 μ/l contains 20 μ g LPS, and blank group gives equal-volume physiological saline; Respectively at the 2nd ?13 days, 15 ?after administration in 35 days 1 hour, mouse is placed in smoking apparatus and carries out fumigation, each 8 mouse, each half root cigarette, 5min/ time, every day 2 times; After last administration 24h, put to death animal, draw materials and carry out Indexs measure.
Iii. Testing index and method:
Bronchoalveolar lavage fluid Arneth's count:
Animal is put to death with after Sodital intraperitoneal injection anesthesia, dissect neck, expose tracheae, with No. 4 half syringe needle promoting the circulation of qi cannulas, with 0.8ml precooling normal saline flushing 3 times, often all over rinsing 3 times back and forth, reclaim irrigating solution (rate of recovery reaches 80%), in 4 DEG C of centrifugal 10min of 1500rpm/min, collect supernatant,-20 DEG C frozen, for cytokines measurement; Contain the PBS re-suspended cell precipitation of 1%BSA with 0.5ml, on five classification differential blood count instrument, carry out Arneth's count.
In bronchoalveolar lavage fluid, the ELISA of relevant inflammatory factors measures:
Get-20 DEG C of frozen irrigating solutions, in thawing on ice, the ELISA kit specification sheets strictly with reference to the corresponding factor operates.
Experimental result
I. on the impact of bronchoalveolar lavage fluid Lymphocyte subset
As shown in Fig. 7 result, compared with model group, significantly can reduce total white blood cells, neutrophil leucocyte number and the eosinophilic granulocyte in irrigating solution after Brusatol, compound 2,39 and positive control drug DEX administration, show as significant anti-mouse COPD inflammatory effect.
Ii. on the impact of mouse bronchoalveolar lavage fluid inflammatory factor secretion situation
As shown in Fig. 8 result, compared with model group, significantly can suppress the secretion of inflammatory factor IL-1 β, IL-6, TNF-α and IL-17 in bronchoalveolar lavage fluid after Brusatol, compound 2,39 and positive control drug DEX administration, prompting has the pulmonary inflammatory effect of remarkable anti-COPD.

Claims (16)

1. the Brusatol derivative shown in general formula I:
Wherein, R 1, R 2, R 4independently be selected from H, the furazan ring replaced,
Wherein, the furazan ring of replacement is selected from or
Wherein, L 1be selected from R 6or-R 6-X 1-Y 1-, wherein, Y 1be connected with O in primer, R 6be selected from C 2-6alkyl, C 2-4oC 2-4alkyl, C 2-6thiazolinyl, C 2-6alkynyl, substituted or unsubstituted phenyl, the substituting group of wherein said phenyl is selected from C 1-6alkyl, X 1be selected from O or NH, Y 1be selected from succinyl or phthaloyl;
L 2be selected from R 7,-R 7-X 2-Y 2-, Y 2, wherein, Y 2be connected with O in primer, R 7be selected from substituted or unsubstituted phenyl, the substituting group of wherein said phenyl is selected from C 1-6alkyl, X 2be selected from O or NH, Y 2be selected from succinyl or phthaloyl;
R 3be selected from OH, OCH 3, replace furazan ring,
The furazan ring wherein replaced is selected from or
Wherein R 9be selected from C 2-6alkyl, C 2-4oC 2-4alkyl, C 2-6thiazolinyl, C 2-6alkynyl, substituted or unsubstituted phenyl, the substituting group of wherein said phenyl is selected from C 1-6alkyl;
L 3be selected from-O-R 10-O-or O, wherein R 10be selected from substituted or unsubstituted phenyl, the substituting group of wherein said phenyl is selected from C 1-6alkyl.
2. compound according to claim 1, is characterized in that, described compound is the compound shown in general formula (Ia):
Wherein, R 3be selected from OH, OCH 3, L 1be selected from-R 6-X 1-Y 1-, wherein, Y 1be connected with O in primer, R 6be selected from C 2-6alkyl, C 2-4oC 2-4alkyl, C 2-6alkynyl ,-o-Ph-(CH 2) n-, wherein n=0,1,2,3,4 ,-m-Ph-(CH 2) n-, wherein n=0,1,2,3,4 ,-p-Ph-(CH 2) n-, wherein n=0,1,2,3,4; X 1be selected from O, Y 1be selected from succinyl or phthaloyl.
3. compound according to claim 1, is characterized in that, described compound is the compound shown in general formula (Ib):
Wherein, R 3be selected from OH, OCH 3, L 2be selected from R 7,-R 7-X 2-Y 2-, Y 2, wherein, Y 2be connected with O in primer, R 7be selected from-o-Ph-(CH 2) n-, wherein n=0,1,2,3,4 ,-m-Ph-(CH 2) n-, wherein n=0,1,2,3,4 ,-p-Ph-(CH 2) n-, wherein n=0,1,2,3,4; X 2be selected from O, Y 2be selected from succinyl or phthaloyl.
4. compound according to claim 1, is characterized in that, described compound is the compound shown in general formula (Ic):
Wherein, L 1be selected from-Y 1-X 1-R 6-, wherein, Y 1be connected with O in primer, R 6be selected from C 2-6alkyl, C 2-4oC 2-4alkyl, C 2-6alkynyl ,-(CH 2) n-o-Ph-, wherein n=0,1,2,3,4 ,-(CH 2) n-m-Ph-, wherein n=0,1,2,3,4 ,-(CH 2) n-p-Ph-, wherein n=0,1,2,3,4, wherein, phenyl connect CH 2all and X 1be connected; X 1be selected from O, Y 1be selected from succinyl or phthaloyl.
5. compound according to claim 1, is characterized in that, described compound is the compound shown in general formula (Id):
Wherein, L 2be selected from-Y 2-X 2-R 7-, wherein, Y 2be connected with O in primer, R 7be selected from-(CH 2) n-o-Ph-, wherein n=0,1,2,3,4 ,-(CH 2) n-m-Ph-, wherein n=0,1,2,3,4 ,-(CH 2) n-p-Ph-, wherein n=0,1,2,3,4, wherein, phenyl connect CH 2all and X 2be connected; X 2be selected from O, Y 2be selected from succinyl or phthaloyl.
6. compound according to claim 1, is characterized in that, described compound is the compound shown in general formula (Ie):
Wherein, R 9be selected from C 2-6alkyl, C 2-4oC 2-4alkyl, C 2-6alkynyl ,-(CH 2) n-o-Ph-, wherein n=0,1,2,3,4 ,-(CH 2) n-m-Ph-, wherein n=0,1,2,3,4 ,-(CH 2) n-p-Ph-, wherein n=0,1,2,3,4, wherein, phenyl connect CH 2all be connected with primer by O.
7. compound according to claim 1, is characterized in that, described compound is the compound shown in general formula (If):
Wherein, L 3be selected from-O-R 10-O-or O,
Wherein, R 10be selected from-(CH 2) n-o-Ph-, wherein n=0,1,2,3,4 ,-(CH 2) n-m-Ph-, wherein n=0,1,2,3,4 ,-(CH 2) n-p-Ph-, wherein n=0,1,2,3,4, wherein, phenyl connect CH 2all be connected with primer by O.
8. compound according to claim 1, is characterized in that, described compound is the compound shown in general formula (Ig):
Wherein, R 1be selected from H, l 1be selected from-R 6-X 1-Y 1-, wherein, Y 1all be connected with O in primer, R 6be selected from C 2-6alkyl, C 2-4oC 2-4alkyl, C 2-6alkynyl ,-o-Ph-(CH 2) n-, wherein n=0,1,2,3,4 ,-m-Ph-(CH 2) n-, wherein n=0,1,2,3,4 ,-p-Ph-(CH 2) n-, wherein n=0,1,2,3,4, wherein, phenyl connect CH 2all and X 1be connected; X 1be selected from O, Y 1be selected from succinyl or phthaloyl.
9. compound according to claim 1, is characterized in that, described compound is the compound shown in general formula (Ii):
Wherein, R 1be selected from H, l 2be selected from-R 7-X 2-Y 2-, Y 2, wherein, Y 2all be connected with O in primer, R 7be selected from-o-Ph-(CH 2) n-, wherein n=0,1,2,3,4 ,-m-Ph-(CH 2) n-, wherein n=0,1,2,3,4 ,-p-Ph-(CH 2) n-, wherein n=0,1,2,3,4, wherein, phenyl connect CH 2all and X 2be connected; X 2be selected from O; Y 2be selected from succinyl or phthaloyl.
10. the nitric oxide donator type Brusatol derivative according to any one of claim 1-9 or its medically acceptable salt, it is characterized in that, compound is selected from:
Compound R 6 X 1 Y 1 1 (CH 2) 3 O CO(CH 2) 2CO 2 (CH 2) 4 O CO(CH 2) 2CO 3 (CH 2) 2CH(CH 3) O CO(CH 2) 2CO 4 (CH 2) 5 O CO(CH 2) 2CO 5 (CH 2) 2O(CH 2) 2 O CO(CH 2) 2CO 6 CH 2C≡CCH 2 O CO(CH 2) 2CO 7 p-PhCH 2 O CO(CH 2) 2CO 8 m-PhCH 2, O CO(CH 2) 2CO 9 (CH 2) 4 O CO-o-PhCO
Compound R 7 X 2 Y 2 10 p-Ph-CH 2 O CO(CH 2) 2CO 11 m-Ph-CH 2 O CO(CH 2) 2CO 12 o-Ph-CH 2 O CO(CH 2) 2CO 13 - O CO(CH 2) 2CO 14 - O CO-o-PhCO 15 p-Ph-CH 2 - -
Compound R 6 X 1 Y 1 16 (CH 2) 3 O CO(CH 2) 2CO 17 (CH 2) 4 O CO(CH 2) 2CO
18 (CH 2) 2CH(CH 3) O CO(CH 2) 2CO 19 (CH 2) 5 O CO(CH 2) 2CO 20 (CH 2) 2O(CH 2) 2 O CO(CH 2) 2CO 21 CH 2C≡CCH 2 O CO(CH 2) 2CO 22 CH 2-p-Ph O CO(CH 2) 2CO 23 CH 2-m-Ph O CO(CH 2) 2CO 24 (CH 2) 4 O CO-o-PhCO
Compound R 7 X 2 Y 2 25 CH 2-p-Ph O CO(CH 2) 2CO 26 CH 2-m-Ph O CO(CH 2) 2CO 27 CH 2-o-Ph O CO(CH 2) 2CO 28 - O CO(CH 2) 2CO 29 - O CO-o-PhCO
Compound R 9 30 CH 2CH 2CH 2 31 CH 2CH 2CH 2CH 2 32 CH 2-p-Ph 33 CH 2-m-Ph
Compound L 3 34 OCH 2-p-PhO 35 OCH 2-m-PhO 36 OCH 2-o-PhO 37 O
11. 1 kinds of pharmaceutical compositions, are made up of the compound described in any one of claim 1-10 of effective dose or its medically acceptable salt and pharmaceutically acceptable carrier or auxiliary material.
12. pharmaceutical compositions according to claim 11, is characterized in that, described pharmaceutical composition is selected from tablet, capsule, pill, injection.
13. pharmaceutical compositions according to claim 11, is characterized in that, described pharmaceutical composition is selected from sustained release preparation, controlled release preparation and various particulate delivery system.
The hydrate of the compound described in 14. any one of claim 1-10 or its medically acceptable salt, salt or the application of prodrug in preparation inflammation and/or immunosuppressive drug.
Compound described in 15. any one of claim 1-10 or its medically acceptable salt, application in preparation inflammation and/or immunologic derangement relative disease medicine.
16. application according to claim 15, wherein inflammation and immunologic derangement relative disease comprise: rheumatoid arthritis, osteoarthritis, rheumatic arthritis, urarthritis, lupus erythematosus syndromes, bronchitis, bursitis, tenosynovitis, psoriasis, eczema, burn, dermatitis, inflammatory bowel, Ke Laoen is sick, gastritis, irritable bowel syndrome, ulcerative colitis, multiple sclerosis, Autoimmune Encephalomyelitis, colorectal carcinoma, arteritis nodosa, thyroiditis, rheumatic fever, gingivitis, periodontitis stomatocace, ephritis, the swelling occurred after damaging, myocardial ischemia, various infectious pneumonia, physics and chemistry pneumonia and allergy pneumonia, chronic obstructive pulmonary disease, asthma, proctalgia fugax and rectum split, liver and gall capsulitis, cholangitis sclerosing cholangitis, primary biliary cirrhosis and cholecystitis.
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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106924245A (en) * 2017-05-08 2017-07-07 广州中医药大学 Purposes of the Brusatol in the medicine for preparing the inhibitor of TLR4/NF κ B paths and treatment IBD
CN111184714A (en) * 2020-02-17 2020-05-22 重庆大学 Application of brucea javanica picrol in preparation of medicines for preventing or treating inflammatory diseases
CN112057446A (en) * 2020-10-19 2020-12-11 重庆市中医院 Application of brucea javanica picrol in preparation of Nrf2/HO-1 signal pathway inhibitor

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106924245A (en) * 2017-05-08 2017-07-07 广州中医药大学 Purposes of the Brusatol in the medicine for preparing the inhibitor of TLR4/NF κ B paths and treatment IBD
CN111184714A (en) * 2020-02-17 2020-05-22 重庆大学 Application of brucea javanica picrol in preparation of medicines for preventing or treating inflammatory diseases
CN112057446A (en) * 2020-10-19 2020-12-11 重庆市中医院 Application of brucea javanica picrol in preparation of Nrf2/HO-1 signal pathway inhibitor

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