CN105713064B - Pentacyclic triterpenoid and its application as people's intestines carboxylesterase inhibitor - Google Patents
Pentacyclic triterpenoid and its application as people's intestines carboxylesterase inhibitor Download PDFInfo
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Abstract
The invention provides a kind of pentacyclic triterpenoid and its application as people's intestines carboxylesterase inhibitor, belong to biomedicine technical field.The pentacyclic triterpenoid has 11 deoxy-glycyrrhetinic acid skeleton structures, its can with it is potent, optionally suppress the activity of people's enteron aisle carboxy-lesterase hypotype 2, and then improve the bioavilability of oral prodrugs, and reduce the intake of enteron aisle Ester.In addition, the inhibitor can also slow down the delayed diarrhoea of Irinotecan initiation.Active determination in vitro finds that such compound suppresses the IC of human carboxylatase 250Up to 0.02 micro- IC for rubbing, suppressing human carboxylatase 150IC with suppressing human carboxylatase 250Ratio is up to 1020, and the security of such compound is preferable, while also having the advantages such as preparation technology is simple, yield is high, points out such compound to show and has a good application prospect.
Description
Technical field
The invention belongs to biomedicine technical field, a kind of pentacyclic triterpenoid is related generally to and its as people's intestines carboxylic
Acid esters potent inhibitor application.
Background technology
The CEs that carboxy-lesterase (CEs) belongs to participation esters medicine metabolism in B races esterase, human body is mainly carboxy-lesterase 1
And carboxy-lesterase 2 (hCE-2) (hCE-1).HCE-1 high expression in liver, and in macrophage, pulmonary epithelial cells, heart, testis
Exist in the tissue such as ball, but express little in intestines and stomach.HCE-2 is deposited in small intestine, colon, kidney, liver, heart and brain tissue
.CEs participates in the hydrolysis metabolism of a variety of endogenous of body and exogenous compounds, and makees jointly with other metabolic enzymes or carrier
With being played an important role in internal metabolism and the reset procedure of esters medicine.CEs hydrolyzables a variety of medicines or ester containing ester bond
Class prodrug, such as angiotensin converting enzyme depressant (Temocapril, Cilazapril, quinapril), (Yi Li is replaced antineoplastic
Health and capecitabine) and arcotic (cocaine, heroin and pethidine) etc..HCE-1 and hCE-2 possesses 48% amino acid
Homologous sequence, but both substrate features have larger difference.Because Irinotecan and heroin show small acyl group end and big hydroxyl
Cardinal extremity, is hCE-2 good substrates, it has been reported that, about 99% Yi Li is replaced in human body duodenum, jejunum, ileum, nephridial tissue
The biological conversion of health is responsible for by hCE-2.
Irinotecan (CPT-11) is camptothecin derivative SN38 pro-drug.At present, due to CPT-11 wide spectrums
Active anticancer makes it be widely used among the chemotherapy of various cancers.But, because its is serious and uncertain diarrhoea
Reaction, makes its clinical practice receive a definite limitation.Diarrhoea caused by CPT-11 includes acute diarrhea and (sent out after administration in 24h
It is raw) and late-onset diarrhea (occurring after administration after 24h) two kinds of [Mol Pharmacol, 2004,65 (6):1336-13431].
3 grades caused by CPT-11 clinical applications and 4 grades of late-onset diarrhea incidences are up to 40%, seriously limit CPT-11 clinic
Use.Domestic and international numerous studies show that the occurrence cause that delayed is suffered from diarrhoea caused by CPT-11 is due to active metabolite SN-
38 excessively gather in enteron aisle.People's enteron aisle carboxy-lesterase, especially hCE-2 has played important function in this pathogenesis.Therefore,
A kind of potent, selectivity hCE-2 inhibitor is developed, conversions of the CPT-11 to SN-38 is reduced, so as to reduce diarrhoea etc.
Toxicity [the J Med Chem, 2009,52 (12) of enteron aisle:3742-37521].
Domestic and international many scholars are directed to studying generation and the alleviation mechanism of Irinotecan adverse reaction, in the hope of improving Yi Li
The security and curative effect used for health.At present, what is the diarrhoea caused by Irinotecan mainly taken is to give Loperamide
Treated.Loperamide reduces enterocinesia by suppressing the contraction of intestinal smooth, reduces intestinal wall nerve endings release acetyl
Choline, directly suppresses to wriggle to reflect and play the effect of alleviating diarrhoea.But it is important to note that diarrhoea is used as the one of body
Defensive measure is planted, is excreted by the larger SN-38 of toxicity, diarrhoea number of times is reduced, for reducing its toxicity to body
The not benefit of essence.And Irinotecan is as SN-38 prodrug, because its activity/active group is closed, its toxicity
Only the 1/1000 of SN-38.Therefore, if the hydrolysis of Irinotecan can be reduced in part (enteron aisle), for reducing to body
The extent of damage is highly beneficial.Therefore, a kind of inhibitor of the potent carboxy-lesterase 2 of safety is developed, for reducing enteron aisle
Local Irinotecan is hydrolyzed into toxic component SN-38 too much, it appears particularly necessary.
Radix glycyrrhizae is the root and rhizome of herbaceos perennial radix glycyrrhizae (Glycyrrhiza urlensis Fisch), and nature and flavor are sweet
Flat, the thoughts of returning home, lung, spleen, stomach are traditional Chinese medicines important simply.The consumption of radix glycyrrhizae is maximum in more than 2000 kinds of Chinese herbal medicines, in
Have that " grass of ten side nine, Cheng Fang " is not said for no grass in medicine composition.It is also that country defends as the radix glycyrrhizae of one of the famous traditional Chinese medicines of China
One of integration of drinking and medicinal herbs food of life portion approval.Modern medicine study shows that the main pharmacological activity material of radix glycyrrhizae is glycyrrhizic acid
(Glycyrrhizic acid) and its aglycon enoxolone (Glycyrrhetinic acid, GA), the former is a class triterpenes
Saponin(e, accounts for the 4%~5% of radix glycyrrhizae root dry weight.Metabolic process of the radix glycyrrhizae acids medicine in human body is clearer, radix glycyrrhizae
Acid intravenous injection after, into liver, in liver cell by the β in lysosome-D-Glucose aldehyde neuraminidase be metabolized to 3- it is mono--grape
After uronic acid enoxolone, enteral is discharged into bile, enoxolone is metabolized to by intestinal flora, is reabsorbed into blood.In recent years, greatly
Quantifier elimination shows, enoxolone and its derivative have antiallergy, analgesia, anti-inflammatory, anticancer, anti AIDS virus, antiulcer, anti-
Oxidation, liver protection isoreactivity.However, enoxolone and its derivative have no report always to the inhibitory action of human carboxylatase,
The report that delayed caused by not alleviating CPT-11 using enoxolone and its derivative is suffered from diarrhoea.In addition, many contains carboxylate
The carboxylesterase metabolism that the oral drugs of key can be often distributed in intestines and stomach, is easily just hydrolyzed into water solubility before absorbed into serum
Larger active medicine, so as to reduce the medication amount of its absorbed into serum, have impact on bioavilability.If taking safety altogether simultaneously
Human carboxylatase inhibitor, can reduce hydrolysis of the prodrug in absorption process, so as to improve its oral administration biaavailability.
The content of the invention
A kind of application of inhibitor the present invention relates to pentacyclic triterpenoid and its as human carboxylatase.The chemical combination
Thing has 11- deoxidations-enoxolone skeleton structure, and it can potently, optionally suppress the activity of human carboxylatase hypotype 2.
Active determination in vitro finds that such compound suppresses the IC of human carboxylatase 250Up to 0.02 it is micro- rub, suppress human carboxylatase 1
IC50IC with suppressing human carboxylatase 250Ratio is up to 1020.The inhibitor can improve the biological utilisation of oral ester prodrug
Spend and extend its metabolic half life, and reduce the intake of enteron aisle Ester.In addition the inhibitor can also alleviate clinical a variety ofization
Treat drug induced delayed diarrhoea.
The invention provides a kind of pentacyclic triterpenoid, the derivative has 11- deoxidations-enoxolone skeleton knot
Structure, its structural formula is as follows：
Wherein, R is any one in methyl, ethyl, propyl group.
A kind of preparation method of pentacyclic triterpenoid, the synthesis of the compound is followed the steps below：
1) zinc powder-hydrochloric acid reduction system is used, 11 carbonyls of enoxolone are removed with high selectivity；Acquisition 11- deoxidations-sweet
Careless hypo acid；
2) with alkyl halide (iodomethane, bromoethane, N-Propyl Bromide) for esterifying reagent, acid binding agent can be potassium carbonate, sodium carbonate, reality
Show the selective esterification of 30 carboxyls, obtain 18 β -11- deoxidations-enoxolone -30- esters；
3) using succinic anhydride as esterifying reagent, DMAP is efficient reaction promoter, is efficiently drawn at 3
Enter butanedioic acid, obtain the β of product 18-(3- carboxyl propionyl groups)-11- deoxidations-enoxolone-30- esters；
The preparation method of the β of step (3) 18-(3- carboxyl propionyl groups)-11- deoxidations-enoxolone-30- esters is specific as follows：
At room temperature, 18 β -11- deoxidations-enoxolone -30- esters, DMAP, succinic anhydride are added to successively
In dichloromethane solution, room temperature reaction, thin plate chromatography (TLC) monitoring reaction are added；After reaction completely, add water, 1M HCl solutions
PH=2-3 is adjusted, ethyl acetate is extracted three times, merge organic phase washing, saturated nacl aqueous solution and wash, anhydrous sodium sulfate drying is steamed
Except solvent, crude product column chromatography obtains the β of product 18-(3- carboxyl propionyl groups)-11- deoxidations-enoxolone-30- esters 3；Each reaction reagent
Consumption in molar ratio be 18 β -11- deoxidations-enoxolone -30- esters：Succinic anhydride：DMAP=1.0:1.5-
3.0:1.5-3.0。
A kind of pentacyclic triterpenoid is as the application of people's intestines carboxylesterase inhibitor, and the compound is used as a kind of people's intestines
The potent inhibitor of road carboxy-lesterase, can potently, optionally suppress the activity of human carboxylatase hypotype 2, it suppresses people's carboxylic
The IC of acid esters enzyme 150IC with suppressing human carboxylatase 250Ratio is up to 1020 times.
A kind of pentacyclic triterpenoid can be prepared containing pentacyclic triterpene as the application of people's intestines carboxylesterase inhibitor
The medicine of compound, the medicine can improve the bioavilability of oral ester prodrug by potent suppression people's enteron aisle carboxy-lesterase, enter
And play effect of ester prodrug synergist.
A kind of pentacyclic triterpenoid can be prepared containing pentacyclic triterpene as the application of people's intestines carboxylesterase inhibitor
The medicine of compound, the medicine can slow down the intestinal absorption of fatty acid material by potent suppression people's enteron aisle carboxy-lesterase, and then
Auxiliary treatment for the metabolic disorder disease such as obesity, diabetes.
A kind of pentacyclic triterpenoid can be prepared containing pentacyclic triterpene as the application of people's intestines carboxylesterase inhibitor
The medicine of compound, the medicine can be alleviated clinical Treated with Chemotherapeutic Drugs thing and caused by potent suppression people's enteron aisle carboxy-lesterase
Delayed diarrhoea.
Described chemotherapeutics, which includes but is not limited to irinotecan hydrochloride, cis-platinum, 5 FU 5 fluorouracil, taxol, topology, to be replaced
Health, methotrexate (MTX), adriamycin, Etoposide, endoxan, BCNU, gemcitabine, epirubicin, vinorelbine etc..
The invention provides application of the described pentacyclic triterpenoid as people's intestines carboxylesterase inhibitor, the derivative
Thing mainly suppresses the carboxy-lesterase 2 of human body intestinal canal height expression, and then can improve the oral bio of prodrug (carboxylester substrates)
Availability：The inhibitor by suppressing the carboxy-lesterase of people's gastrointestinal system, can subtract when being taken altogether with prodrug (carboxylester substrates)
The stomach and intestine hydrolysis of few prodrug (carboxylester substrates) and first pass effect, improve its bioavilability.
The invention provides another application of described pentacyclic triterpenoid as people's intestines carboxylesterase inhibitor,
The carboxylesterase inhibitor can weaken diarrhoea caused by antineoplastic Irinotecan：The inhibitor and Irinotecan antineoplastic
Share, suppress the hydrolysis of Irinotecan in the gastrointestinal tract, and then reduce SN-38 SN-38 in gastrointestinal system
Local exposed amount, slows down its caused delayed diarrhoea serious adverse reaction.
The carboxylesterase inhibitor that the present invention is provided causes diarrhoea with improving prodrug oral bio profit in decrease antineoplastic
Application in terms of expenditure, the constituents are made formulation after being mixed as monomer or compound with conventional auxiliary material and used, also can be with resisting
Tumour medicine is used after being mixed by different proportion as pharmaceutical composition.
The present invention relates to a kind of pentacyclic triterpenoid and its application as human carboxylatase inhibitor, advantage is such as
Under：
1st, it is cheap and easy to get：The novel carboxylic acid esterase inhibitor that the present invention is provided is using cheap enoxolone as raw material, through changing
Learn synthesis to obtain, synthesis technique is simple and easy to apply, and yield is higher.
2nd, high inhibitory activity：The novel carboxylic acid esterase inhibitor presses down in people's tissue microsomal to the half of carboxy-lesterase 2
Concentration IC processed50Up to nM grades.
3rd, high selectivity：Active determination in vitro finds that such compound suppresses the IC of carboxy-lesterase 150With suppressing carboxy-lesterase
2 IC50Ratio is up to 1020.
4th, high security：The novel carboxylic acid esterase inhibitor has good security, its Mouse oral LD50More than 1g/
kg。
Brief description of the drawings
The general structure of Fig. 1 Enoxolone derivatives；
Fig. 2 (3- carboxyl propionyl groups) -11- deoxidations-enoxolone -30- ethyl esters1H-NMR spectrum；
Fig. 3 (3- carboxyl propionyl groups) -11- deoxidations-enoxolone -30- ethyl esters13C-NMR spectrograms；
The synthetic route of Fig. 4 (3- carboxyl propionyl groups) -11- deoxidations-enoxolone -30- ethyl esters；
Fig. 5 (3- carboxyl propionyl groups) -11- deoxidations-enoxolone -30- ethyl esters suppress hCE2 suppression curve；
Embodiment
The following examples will be further described to the present invention, but not thereby limiting the invention.
Equipment and its model of the present invention：Fluorescent emission/excitation spectrum is by SynergyH1 global function micropores
Board detector detection is completed；1H-NMR spectrum and13C-NMR spectrograms are examined by nuclear magnetic resonance chemical analyser (Avance II 400MHz)
Survey and complete.
Embodiment 1
The synthesis of (3- carboxyl propionyl groups) -11- deoxidations-enoxolone -30- carboxylates
(1) synthesis of compound 1 (11- deoxidations-enoxolone)
Room temperature, by 18 β-enoxolone (235.3mg, 0.5mmol), zinc powder (526.5mg, 16.2mmol) be added to Isosorbide-5-Nitrae-
In dioxane (9mL) solution, cooling reaction system is slowly added dropwise concentrated hydrochloric acid (1.45 mL), adds holding system to 8-12 DEG C
8-12 DEG C of reaction, thin plate chromatography (TLC) monitoring reaction.After reaction completely, solvent is removed under reduced pressure, add water (25mL), dichloromethane
Three times (30mL × 3) are extracted, merges organic phase saturated nacl aqueous solution and washes (20mL × 1), anhydrous sodium sulfate drying is evaporated off molten
Agent, crude product column chromatography (petrol ether/ethyl acetate=20/1-5/1) obtains compound 1, white solid, yield 65-75%.
(2) synthesis of compound 2 (11- deoxidations-enoxolone -30- ethyl esters)
Room temperature, third is added to by compound 1 (167.0mg, 0.37mmol), Anhydrous potassium carbonate (51.1mg, 0.37mmol)
In the mixed solution of ketone (10mL) and tetrahydrofuran (5mL), bromoethane (41 μ L, 0.55mmol) is added dropwise after stirring 5min, adds
It is warming up to 35 DEG C of reactions, thin plate chromatography (TLC) monitoring reaction.After reaction completely, solvent is removed under reduced pressure, add water (20mL), dichloro
Methane extracts three times (25mL × 3), merges organic phase saturated nacl aqueous solution and washes (15mL × 1), anhydrous sodium sulfate drying is evaporated off
Solvent, crude product column chromatography (petrol ether/ethyl acetate=50/1-10/1) obtains compound 2, white solid, yield 85-95%.
(3) synthesis of compound 3 ((3- carboxyl propionyl groups) -11- deoxidations-enoxolone -30- ethyl esters)
Room temperature, successively by compound 2 (89.0mg, 0.18mmol), DMAP (45.0mg, 0.36mmol),
Succinic anhydride (91.9mg, 0.92mmol) is added in dichloromethane (2mL) solution, adds room temperature reaction, thin plate chromatography (TLC)
Monitoring reaction.After reaction completely, add water (10mL), and 1M HCl solutions adjust pH=2-3, and ethyl acetate extracts three times (25mL × 3),
Merge organic phase washing (15mL × 1), saturated nacl aqueous solution to wash (15mL × 1), solvent is evaporated off, slightly in anhydrous sodium sulfate drying
Product column chromatography (methylene chloride/methanol=100/1-20/1 gradient elutions) obtains compound 3, white solid, yield 70-80%.
The NMR spectrum of product is specific as follows：
1H NMR(400MHz,CDCl3)δ5.26(s,1H),4.61-4.43(m,1H),4.29-3.98(m,2H),2.69-
4.24(m,4H),2.03-1.82(m,6H),1.80-1.74(m,1H),1.69-1.47(m,7H),1.47-1.38(m,1H),
1.37-1.21(m,8H),1.14(s,3H),1.12(s,3H),1.09-0.99(m,2H),0.96(s,6H),0.83-0.87(m,
8H),0.78(s,3H)；13C NMR(101MHz,CDCl3)δ177.2,171.9,144.6,122.4,81.6,60.0,55.3,
48.2,47.56,44.1,42.8,41.5,39.8,38.3,38.2,37.8,36.8,32.6,31.9,31.3,29.4,28.8,
28.5,28.2,28.0,27.0,26.2,25.9,23.5,18.2,16.8,16.7,15.5,14.3.
Embodiment 2.
Qualitative assessment of (3- the carboxyl propionyl groups) -11- deoxidations-enoxolone -30- carboxylates to the rejection ability of carboxy-lesterase 2
Probe reaction is metabolized as with the hydrolysis to fluorescein(e) diacetate, by people's hepatomicrosome incubated in vitro system, surveyed
Determine the IC that enoxolone and its derivative suppress to carboxy-lesterase 250：
A.200 in microlitre In vitro metabolism reaction system, containing the phosphate buffer that pH is 7.4, people's hepatomicrosome albumen is dense
Spend for 2 μ g/ml, inhibitor final concentration scope is 0.001 μM -100 μM, shakes and incubates 10 minutes in advance under the conditions of 37 DEG C；
B. substrate (10 μM of final concentration), initial action are added into reaction system；After being reacted 30 minutes under the conditions of 37 DEG C,
200 μ l acetonitriles are added, acutely after concussion, terminating reaction；
C. high speed freezing centrifuge is used, under conditions of 20,000 × g, the above-mentioned system of high speed centrifugation takes after 5 minutes
Clearly, ELIASA detection and analysis is carried out；Metabolism hydrolysate is quantitatively detected.
Embodiment 3.
Qualitative assessment of (3- the carboxyl propionyl groups) -11- deoxidations-enoxolone -30- carboxylates to the rejection ability of carboxy-lesterase 1
Probe reaction is metabolized as with the hydrolysis to fluorescein(e) diacetate, by people's hepatomicrosome incubated in vitro system, surveyed
Determine the IC that enoxolone and its derivative suppress to carboxy-lesterase 150：
A.200 in microlitre In vitro metabolism reaction system, containing the phosphate buffer that pH is 7.4, people's hepatomicrosome albumen is dense
Spend for 2 μ g/ml, inhibitor final concentration scope is 0.001 μM -120 μM, shakes and incubates 10 minutes in advance under the conditions of 37 DEG C；
B. substrate (10 μM of final concentration), initial action are added into reaction system；After being reacted 30 minutes under the conditions of 37 DEG C,
200 μ l acetonitriles are added, acutely after concussion, terminating reaction；
C. high speed freezing centrifuge is used, under conditions of 20,000 × g, the above-mentioned system of high speed centrifugation takes after 5 minutes
Clearly, ELIASA detection and analysis is carried out；Metabolism hydrolysate is quantitatively detected.
The suppression of the enoxolone of table 1 and its derivative to carboxy-lesterase
Enoxolone and its derivative show good inhibitory activity to hCE2, and the half-inhibition concentration of each compound is such as
Shown in table 1.From obtained experimental data as can be seen that transformation enoxolone (3 acylated hydroxies, removing 11 carbonyls, 30 carboxyls
Esterification) its inhibitory activity to hCE2 can be significantly improved, while significantly increasing the selectivity to hCE2.(3- carboxylics third after transformation
Acyl group) -11- deoxidations-enoxolone -30- ethyl esters are 3463 times of enoxolone to hCE2 inhibitory activity, suppress carboxy-lesterase 1
IC50IC with suppressing carboxy-lesterase 250Ratio is up to 1020.5.
Embodiment 4.
The Mouse oral acute toxicity of (3- carboxyl propionyl groups) -11- deoxidations-enoxolone -30- carboxylic acid, ethyl esters is assessed
Choose Balb/c mouse (being purchased from Dalian Medical Univ's Experimental Animal Center), male and female half and half, body weight 19-22g.Will be small
Mouse is grouped at random, every group 20, male and female half and half.Test compound is suspended in 0.5%CMC-Na, and concentration is 1g/L.Experimental group
Including test compound various dose group (0.1~1g/kg) and 0.5%CMC-Na blank control groups.After Continuous Observation administration
The behavior state of mouse carried out gross anatomy to different dosing group mouse in the 14th day and observed internal organ situation up to 14 days.
There was only 4 death in maximum dose group (1g/kg) administration group mouse, other dosage groups are without dead example.Dead individuals are carried out
Dissection does not find the obvious lesion of the major organs such as liver, kidney.Test result indicates that Mouse oral (3- carboxyl propionyl groups) -11- is de-
The LD50 values of oxygen-enoxolone -29- carboxylic acid, ethyl esters are more than 1.0g/kg, point out the novel carboxylic acid esterase inhibitor to have well
Security.
Embodiment 5.
(3- carboxyl propionyl groups) -11- deoxidations-enoxolone -30- carboxylic acid, ethyl esters are to slowing down Induced by High Fat Diet rat trophism
Fat research
45 Adult SD male rats are chosen, being formulated made higher fatty acid, high heat food using D12492 is limited
System feeding, every mouse 13g in the 1st week increases weekly 2g, untill the 6th week later.It is daily to raise point 2 supplies, after eating up not
Add again, free water.Rejected in the rat that increased weight is come to rear 1/3 after 3 weeks as Diet resistant rat.Remaining rat
It is divided into chow diet group, high fat diet model group and (3- carboxyl propionyl groups) -11- deoxidation-enoxolones-according to random group forming criterion
30- carboxylic acid, ethyl esters administration group (hereinafter referred to as deoxy-glycyrrhetinic acid group), every group of 10 rats.Chow diet group only awards normal feeding
Material, high fat diet group continue to give high fat diet, and deoxy-glycyrrhetinic acid group is in by 300 mg/kg (daily, three times) dosage mouthful
Take gavage and give (3- carboxyl propionyl groups) -11- deoxidations-enoxolone -30- carboxylic acid, ethyl esters.(i.e. 4-7 is tested within 4 weeks for continuous operation
Week), and respectively to body weight and Lee ' s indexes (length of the rat from nose to anus is measured after anesthesia, and [Lee ' s refer to according to formula
The body length (cm) of number=body weight (g) 1/3 × 103/] calculate Lee ' s indexes) observed.
The rat body weight of table 2. and the observation of Lee ' s indexes
Experimental result confirms that giving rat (3- carboxyl propionyl groups) -11- deoxidations-enoxolone -30- carboxylic acid, ethyl esters can have
Rat body weight excessively increases caused by effect alleviates high fat diet, points out it right by suppressing the CES2 of high expression in enteron aisle
Lipid compounds homeostasis is adjusted, and suppresses fat generation.
Embodiment 6.
(3- carboxyl propionyl groups) -11- deoxidations-enoxolone -30- carboxylic acid, ethyl esters are to slowing down diarrhea of mouse caused by Irinotecan
Research
18 Balb/c mouse are randomly divided into 3 groups：Normal group, Irinotecan Diarrhea Model group and (3- carboxylic propionyl
Base) -11- deoxidations-enoxolone -30- carboxylic acid, ethyl esters+Irinotecan Diarrhea Model group (hereinafter referred to as deoxy-glycyrrhetinic acid group),
Every group each 6.Observation Normal group, Diarrhea Model group and the diarrhoea status of deoxy-glycyrrhetinic acid group mouse, and to small respectively
Mouse intestinal tissue carries out histotomy inspection.Diarrhea Model group presses Trifan methods (Cancer Res 2002；62:5778-
84.), Diarrhea Model, which is used, is injected intraperitoneally Irinotecan (100mg/kg/d), continuous use 3 days (d), and late-onset diarrhea is in the 3rd
It occurred, in the 4th day most serious.Deoxy-glycyrrhetinic acid group is in being injected first 3 days CPT-11,1 time a day, gavage 100mg/
Kg, remaining two groups prevent the emergent influence caused to mouse with isometric distilled water gavage.Normal group tail vein injection
Isometric physiological saline.Normal group, Diarrhea Model group and the diarrhoea status of deoxy-glycyrrhetinic acid group mouse are observed respectively, in
Put to death mouse within 7th day, carry out intestinal tissue materials, tissue section strain inspection.
Inject CPT-11 after the 9th day put to death mouse, away from fetched at ileocaecal sphineter 5cm intestines 3cm, caecum 1cm, anus supreme 7~
9cm takes colon 3cm, and 10% formaldehyde is fixed, in case om observation.Conventional H E dyeing observation each group mouse intestinal submucosa tissue knots
Structure changes；According to Chiu intestinal mucosal injuries methods of marking (Arch Surg 1970；101:478-83), to intestinal mucosal injury degree
It is classified：1 grade：Normal colonic mucosa fine hair；2 grades：Subcutaneous clearance expands, and generally on intestinal villus top, often there is upper dermathemia；
3 grades：Subcutaneous clearance is expanded, and departs from moderate epithelial layer from intestinal mucosa lamina propria；4 grades：In the bulk of intestinal mucosa side
Skin departs from, and most of intestinal villus top polishes；5 grades：Intestinal villus polishes, telangiectasis, the intrinsic confluent monolayer cells of intestinal mucosa
Constitute increase；6 grades：Intestinal mucosa lamina propria digests and decomposed, and bleeding and ulcer occurs.
Intestinal mucosal injury degree is estimated as shown in table 3, experimental result is shown, diarrhoea degree is remarkably decreased, deoxidation
Enoxolone group intestinal mucosa damaged condition is slight compared with model group.The confirmation of above experimental result, (3- carboxyl propionyl groups) -11- deoxidations-sweet
Careless hypo acid -30- carboxylic acid, ethyl esters can not only reduce the mouse late-onset diarrhea of CPT-11 inductions and the degree of intestinal mucosal injury, also
The generation of diarrhoea can preferably be suppressed.Therefore, (3- carboxyl propionyl groups) -11- deoxidations-enoxolone -30- carboxylates are lured CPT-11
The mouse late-onset diarrhea of hair has certain prevention effect.
The mouse caecum intestinal mucosal injury grading n (%) of table 3.
Claims (7)
1. a kind of pentacyclic triterpenoid, it is characterised in that：The compound is that 11- deoxidations-enoxolone -3-O- succinyls spread out
Biological carboxylate, its structural formula is as follows：
Wherein, R is any one in methyl, ethyl, propyl group.
2. a kind of pentacyclic triterpenoid preparation method as claimed in claim 1, it is characterised in that the synthesis of the compound
Follow the steps below：
1) zinc powder-hydrochloric acid reduction system is used, 11 carbonyls of enoxolone are optionally removed；Obtain 11- deoxidations-radix glycyrrhizae time
Acid；
2) using alkyl halide as esterifying reagent, acid binding agent is potassium carbonate, sodium carbonate, realizes the selective esterification of 30 carboxyls, obtains
Obtain 11- deoxidations-enoxolone -30- esters；
3) using succinic anhydride as esterifying reagent, DMAP is reaction promoter, introduces butanedioic acid at 3, is produced
The carboxylate of thing 11- deoxidations-enoxolone -3-O- succinyl derivatives,
The alkyl halide is iodomethane, bromoethane or N-Propyl Bromide.
3. according to the pentacyclic triterpenoid preparation method described in claim 2, it is characterised in that step (3) 11- takes off
The preparation method of the carboxylate of oxygen-enoxolone -3-O- succinyl derivatives is specific as follows：
At room temperature, 11- deoxidations-enoxolone -30- esters, DMAP, succinic anhydride are added to dichloromethane successively
In solution, room temperature reaction, thin plate chromatography (TLC) monitoring reaction are added；After reaction completely, add water, 1M HCl solutions adjust pH=2-
3, ethyl acetate is extracted three times, is merged organic phase washing, saturated nacl aqueous solution and is washed, solvent is evaporated off, slightly in anhydrous sodium sulfate drying
Product column chromatography obtains product (3- carboxyl propionyl groups) -11- deoxidations-enoxolone -30- esters；
The consumption of each reaction reagent is 11- deoxidations-enoxolone -30- esters in molar ratio：Succinic anhydride：DMAP
=1.0:1.5-3.0:1.5-3.0.
4. a kind of application of pentacyclic triterpenoid as claimed in claim 1 in people's intestines carboxylesterase inhibitor is prepared,
It is characterized in that：The compound optionally suppresses human carboxylatase hypotype 2 as a kind of inhibitor of people's enteron aisle carboxy-lesterase
Activity, its suppress human carboxylatase 1 IC50IC with suppressing human carboxylatase 250Ratio is up to 1020 times.
5. according to the pentacyclic triterpenoid described in claim 4 prepare people's intestines carboxylesterase inhibitor in application, its
It is characterised by preparing the medicine containing pentacyclic triterpenoid, the medicine improves oral esters by suppressing people's enteron aisle carboxy-lesterase
The bioavilability of prodrug, plays effect of ester prodrug synergist.
6. according to the pentacyclic triterpenoid described in claim 4 prepare people's intestines carboxylesterase inhibitor in application, its
It is characterised by preparing the medicine containing pentacyclic triterpenoid, the medicine slows down fatty acid by suppressing people's enteron aisle carboxy-lesterase
The intestinal absorption of material, the auxiliary treatment for fat, diabetes.
7. according to the pentacyclic triterpenoid described in claim 4 prepare people's intestines carboxylesterase inhibitor in application, its
It is characterised by preparing the medicine containing pentacyclic triterpenoid, the medicine is alleviated clinical many by suppressing people's enteron aisle carboxy-lesterase
Plant delayed diarrhoea and other side effects caused by chemotherapeutics.
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| PCT/CN2015/000327 WO2016086496A1 (en) | 2014-12-05 | 2015-05-13 | Pentacyclic triterpenoid compound and use thereof as human intestinal carboxylesterase (ces) inhibitor |
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| CN112176018B (en) * | 2020-08-28 | 2022-09-13 | 河北仁心药业有限公司 | Method for preparing glycyrrhetinic acid and derivatives thereof based on honey-fried licorice root and application thereof |
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| GB1475075A (en) * | 1974-09-11 | 1977-06-01 | Biorex Laboratories Ltd | Derivatives of glycyrrhetinic and oleanolic acids |
| CN101200488A (en) * | 2007-12-07 | 2008-06-18 | 北京润德康医药技术有限公司 | Novel biogastrone acid derivatives, preparation method and medical uses thereof |
| CN101508719A (en) * | 2009-03-31 | 2009-08-19 | 南京工业大学 | 11-deoxidized biogastrone acid-3-0-succinic acid half-ester rare earth salt and synthesis thereof |
| CN101899081A (en) * | 2009-05-31 | 2010-12-01 | 江苏正大天晴药业股份有限公司 | Synthetic method of glycyrrhetinic acid ester derivative and deoxyglycyrrhetinic acid ester compound |
-
2014
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|---|---|---|---|---|
| GB1475075A (en) * | 1974-09-11 | 1977-06-01 | Biorex Laboratories Ltd | Derivatives of glycyrrhetinic and oleanolic acids |
| CN101200488A (en) * | 2007-12-07 | 2008-06-18 | 北京润德康医药技术有限公司 | Novel biogastrone acid derivatives, preparation method and medical uses thereof |
| CN101508719A (en) * | 2009-03-31 | 2009-08-19 | 南京工业大学 | 11-deoxidized biogastrone acid-3-0-succinic acid half-ester rare earth salt and synthesis thereof |
| CN101899081A (en) * | 2009-05-31 | 2010-12-01 | 江苏正大天晴药业股份有限公司 | Synthetic method of glycyrrhetinic acid ester derivative and deoxyglycyrrhetinic acid ester compound |
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| Synthesis and Biological Activities of Substituted Glycyrrhetic Acids;John S. Baran等;《Journal of Medicinal Chemistry》;19741231;第17卷(第2期);第184-191页 * |
| 甘草酸、甘草次酸衍生物的合成及生物活性研究进展;雍建平;《时珍国医国药》;20121231;第23卷(第6期);第1495-1498页 * |
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