CN101570528A

CN101570528A - Glycyrrhizin derivatives and preparation and use thereof

Info

Publication number: CN101570528A
Application number: CNA200910148227XA
Authority: CN
Inventors: 吕秋军
Original assignee: Individual
Current assignee: Individual
Priority date: 2009-06-18
Filing date: 2009-06-18
Publication date: 2009-11-04
Anticipated expiration: 2029-06-18
Also published as: CN101570528B; WO2010145207A1

Abstract

The invention relates to Glycyrrhizin derivatives and preparation and use thereof, in particular to a compound in formula I: wherein, n is 0 or 1; R1 is C3-8 cycloalkyl; and R2 is H, as well as isomers, racemes, antimers, diastereomers, antimer concentrates, solvates, prodrugs and esters thereof. The invention further relates to a preparation method of the compound in the formula I, a pharmaceutical composition containing the compound in the formula I, and use of the compound in the formula I for preparing drugs for treating and/or preventing cerebral ischemic diseases, neurodegenerative diseases and inflammatory diseases. The compound in the formula I can be used for effectively treating and/or preventing the diseases described in the invention.

Description

Glycyrrhizin derivatives and method for making thereof and purposes

Technical field

The present invention relates to class glycyrrhizin derivatives and preparation method thereof, and the pharmaceutical composition that contains them.Glycyrrhizin derivatives of the present invention can be used for the treatment of and/or prevention of brain ischemic disease, nerve degenerative diseases and inflammatory diseases effectively.

Background technology

Contain a large amount of flavones compositions in the Radix Glycyrrhizae, comprising Liquiritigenin (Liquiritigenin), promptly 4 ', 7-Dihydroxyflavanone (Chinese name: 4 ', 7-dihydroxyl-flavanone), be called again 7-Hydroxy-2-(4-hydroxy-phenyl)-chroman-4-one (Chinese name: 7-hydroxyl-2-(4-hydroxyl-phenyl)-chroman-4-on-), its structural formula is as follows:

Bibliographical information, Liquiritigenin have anti-acute liver damage (Chem Biol Interact, 2006,161 (2): 125-138), suppress XOD (Cell mol Life Sci, 2000,57 (3): 500-505) isoreactivity.The inventor discloses Liquiritigenin and has had the new activity of obvious treatment nerve degenerative diseases in the patent of application before this, and not having simultaneously significantly stimulates breast cancer cell proliferation function (application number 200710163547.3).

Estrogen receptor (ER) mainly comprises ER α and two kinds of hypotypes of ER β of exercising various physiological functions, wherein ER α is main relevant with sex, and the relation of ER β and mood regulation and cognitive behavior more close (Brain Research Reviews, 2008,57:309-320).Therefore, find that ER β subtype agonist has great potential in aspect medicines such as exploitation hypermnesis, neuroprotective.Liquiritigenin is selective estrogen receptor (ER) β subtype agonist (Mol CellEndocrinol.2008,283 (1-2): 49-57), also have tangible anti-inflammatory action (Br JPharmacol.2008,154 (1): 165-73).Laid-open U.S. Patents has reported that also Liquiritigenin is the β subtype agonist (number of patent application US 2008/0319051A1, US 2009/0042818A1) of selectivity ER subsequently.

Because the Liquiritigenin characteristics of molecular structure has two phenolic hydroxyl groups, instability on the parent nucleus, easily by the esterification of II phase metabolic enzyme, metabolic stability is relatively poor relatively for phenolic hydroxyl group on the parent nucleus, and retention time is short in vivo, for reaching stable Plasma Concentration, need to repeat every day 4-5 administration clinically.Above-mentioned these deficiencies have limited the application of Liquiritigenin as medicine.We are optionally by modifying synthetic new derivative to the side-chain structure of Liquiritigenin, strong ER β agonist activity is found in screening, has stronger pharmacologically active, metabolic stability also is enhanced, obviously reduce the medication number of times, the conformability of patient's medication strengthens, and helps gerontal patient's medication.

In existing document, do not see the present invention's the Liquiritigenin ether or the report of ester derivative as yet, and these derivatives are at treatment or prevention of brain ischemic disease, nerve degenerative diseases, and the purposes of treatment ulcerative colitis, rheumatoid arthritis.

Cerebral ischemia diseases accounts for the overwhelming majority (60-80%) of cerebrovascular disease, comprises cerebral thrombosis, cerebral embolism, lacunar infarction and transient ischemic attack.In China, cerebral ischemia diseases occupies three big cause of the death first places more than heart trouble and cancer.Cerebral thrombosis, cerebral embolism have the characteristics of high incidence, high disability rate and high relapse rate, cause a large amount of group of handicapped, have brought heavy society and economical load.Pharmacological agent comprises vasodilation, anticoagulation, platelet aggregation-against and thromboembolism treatment.But the result of treatment of overall ischemic encephalopathy is also undesirable, still presses for the new medicine of invention.

Nerve degenerative diseases is meant the disease that the sex change of nervous tissue chronic degenerative is produced, mainly contain Parkinson's disease (Parkinsonism), alzheimer's disease etc., medicine has anticholinesterase (cholinesterase inhibitor at present, AchEI) and the nmda receptor conditioning agent, clinical effectiveness is not good.

Ulcerative colitis is a kind of of inflammatory bowel, and it is rectum and colon chronic nonspecific inflammation disease.Therapeutic purpose control inflammation exactly.The anti-inflammatory drug kind is more, comprise sulfasalazine, cortin, immunosuppressor, metronidazole etc., but above-mentioned generally treatment can only be alleviated a part of symptom, and therefore easily recurrence after the drug withdrawal needs new role mechanism and medicine evident in efficacy.

Rheumatoid arthritis (rheumatoid arthritis) be a kind of be the chronic systemic autoimmune disease of feature with the arthrosynovitis.Synovitis is outbreak repeatedly lastingly, the destruction that can cause intraarticular cartilage and bone, and joint function disturbance, in addition maimed.Medicine comprises non-steroidal anti-inflammatory drugs and immunosuppressor Rheumatrex etc., and result of treatment is all not good.

Summary of the invention

In order to overcome the deficiencies in the prior art, the objective of the invention is to be to provide a class Liquiritigenin ethers or an ester derivative; Another object of the present invention is to provide the preparation method of preparation Liquiritigenin ethers or ester derivative raceme.From formula (I) as can be known, Liquiritigenin ethers or ester derivative C2 are chiral carbon, and by chiral separation, the preparation enantiomorph detects the exciting ER β of raceme or enantiomorph isoreactivity.Therefore, a further object of the present invention is to provide the method for separating Liquiritigenin ether derivative enantiomorph; Another purpose of the present invention is to detect the exciting ER β and the neuroprotective activity of Liquiritigenin ether derivative raceme; A further object of the present invention is to detect the exciting ER β and the neuroprotective activity of Liquiritigenin ether derivative levo form; Another purpose of the present invention is to detect the exciting ER β and the neuroprotective activity of Liquiritigenin ether derivative dextrorotatory form; A further object of the present invention is to provide the pharmaceutical composition that contains one or more Liquiritigenin ether derivative racemies or enantiomorph; Another purpose of the present invention is to provide the raceme of Liquiritigenin ether derivative or enantiomorph to prevent and treat cerebrovascular disease in preparation, especially the purposes in the cerebral ischemia diseases medicine; Another object of the present invention is to provide the raceme or the purposes of enantiomorph in preparation treatment nerve degenerative diseases medicine of Liquiritigenin ether derivative; A further object of the present invention is to provide the raceme or the purposes of enantiomorph in preparation treatment ulcerative colitis medicine of Liquiritigenin ether derivative; Another object of the present invention is to provide the raceme or the purposes of enantiomorph in preparation treatment rheumatoid arthritis medicine of Liquiritigenin ether derivative.The inventor finds that novel compounds of formula I provided by the invention is that Liquiritigenin ethers or ester derivative have the pharmacological activity that makes us expecting, the present invention is based on this discovery and is accomplished.

Summary of the invention:

First aspect present invention provides formula I compound:

Wherein:

N is 0 or 1;

R1 is the C3-8 cycloalkyl;

R2 is H,

And isomer, raceme, enantiomorph, diastereomer, enantiomorph enriched substance, solvate, prodrug and ester.

Compound according to first aspect present invention:

Wherein:

N is 0;

R1 is selected from: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, suberyl;

R2 is H,

Compound according to first aspect present invention:

Wherein:

N is 1;

R1 is selected from: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, suberyl;

R2 is H,

According to the compound of first aspect present invention, it is selected from formula II and formula III compound, and specifically, it is selected from:

[being 2-(4-cyclopentyloxy-phenyl)-7-hydroxyl-chroman-4-on-, 2-(4-Cyclopentyloxy-phenyl)-7-hydroxy-chroman-4-one]; With

[being 4-(7-hydroxyl-4-oxo-chroman-2-yl)-benzyl ring pentane manthanoate, Cyclopentanecarboxylic acid4-(7-hydroxy-4-oxo-chroman-2-yl)-phenyl ester],

According to the compound of first aspect present invention, it is selected from formula IIa, formula IIb, formula III a and formula III b compound, and specifically, it is selected from:

And enantiomorph enriched substance, solvate, prodrug and ester.

According to the compound of first aspect present invention, it is:

Basically optically pure

Basically optically pure

Basically optically pure Or

Basically optically pure

And solvate.

Second aspect present invention provides the method for preparing each described compound of first aspect present invention.

According to the method for second aspect present invention, particularly can be used to prepare wherein n and be the method for 1 compound (particularly formula III compound), this method may further comprise the steps:

A) make p-Hydroxybenzaldehyde and formula

Shown in compound reaction, obtain formula

Compound;

B) make formula

Compound and formula Hal-CH ₂OCH ₃The compound reaction obtains formula Compound;

C) make the reaction of step a) and step b), obtain formula

Compound;

D) make the reaction product hydrolysis of step c), obtain formula Compound; With

E) make the reaction product of step d) carry out cyclization and obtain formula I compound, and the optional step of formula I compound being carried out chiral separation,

Wherein: Hal represents halogen for example fluorine, chlorine, bromine, iodine; The definition of n and R1 is with first aspect present invention each described (especially, n wherein is 1).

In an embodiment of second aspect present invention method, the method for preparation formula II compound is provided, it may further comprise the steps:

A) in the presence of alkali (for example salt of wormwood), make formula

With formula The compound reaction obtains formula

Compound;

B) in the presence of alkali (for example salt of wormwood), make formula

Compound and methoxychlor methane reaction obtain formula

Compound;

C) in the presence of alkali (for example sodium hydroxide), make the reaction of step a) and step b), obtain formula

Compound;

D) in The suitable solvent (for example ethanol), make the reaction product hydrolysis of step c) in the presence of (for example concentrated hydrochloric acid), obtain formula in acid

Compound; With

E) make the reaction product of step d) carry out cyclization (for example in ethanol, in the presence of sodium acetate, at high temperature for example under reflux temperature) and obtain formula II compound; And the optional step of formula II compound being carried out chiral separation.

With reference to the method for second aspect present invention, and/or, can also synthesize other formula I compound, for example formula IIa, formula IIb, formula III compound etc. in conjunction with prior art disclosed method (for example referring to US2008/0319051A1).

A third aspect of the present invention relates to a kind of pharmaceutical composition, and it comprises each described formula I compound of first aspect present invention and optional one or more pharmaceutically acceptable carriers or the vehicle that treats and/or prevents significant quantity.

Fourth aspect present invention relate to each described formula I compound of first aspect present invention preparation be used for the treatment of and/or the medicine of prevention of brain ischemic disease in purposes.

Fifth aspect present invention relates to each described formula I compound of first aspect present invention and is used for the treatment of and/or prevents purposes in the medicine of nerve degenerative diseases in preparation.In an embodiment of fifth aspect present invention, provide first aspect present invention each described formula I compound to be used for the treatment of and/or to prevent purposes in the medicine of Parkinson's disease (Parkinsonism) or alzheimer's disease in preparation.

Sixth aspect present invention relates to each described formula I compound of first aspect present invention and is used for the treatment of and/or prevents purposes in the medicine of inflammatory diseases in preparation.In an embodiment of sixth aspect present invention, provide first aspect present invention each described formula I compound preparation be used for the treatment of and/or the medicine of prevention of ulcerative colitis or rheumatoid arthritis in purposes.

Seventh aspect present invention relates to the purposes of each described formula I compound of first aspect present invention in the preparation medicine, and described medicine is used for the next item down or multinomial:

I) agonist of erss hypotype (ER β);

Ii) hypermnesia agent;

Iii) neuroprotective; With

Iv) analgesic agent.

Eighth aspect present invention relates to a kind of method that treats and/or prevents disease in the Mammals of needs is arranged, this method comprises each the described formula I compound of first aspect present invention that treats and/or prevents significant quantity to the administration that needs are arranged, and wherein said disease is selected from: cerebral ischemia diseases, nerve degenerative diseases (such as but not limited to Parkinson's disease (Parkinsonism) or alzheimer's disease), inflammatory diseases (such as but not limited to ulcerative colitis or rheumatoid arthritis).

Detailed Description Of The Invention:

All documents that the present invention quoted from, their full content is incorporated this paper by reference into, and if the expressed implication of these documents and the present invention when inconsistent, be as the criterion with statement of the present invention.In addition, various terms and phrase that the present invention uses have the general sense of well known to a person skilled in the art, nonetheless, the present invention still wishes at this more detailed description and interpretation to be made in these terms and phrase, term of mentioning and phrase are as the criterion with the implication that the present invention was explained if any inconsistent with known implication.

Term " halogen ", " Hal " or " halo " are meant fluorine, chlorine, bromine and iodine.

The term that is adopted among the present invention " alkyl ", " alkenyl " and " alkynyl " have general sense well known in the art, they are hydrocarbyl groups of straight or branched, such as but not limited to methyl, ethyl, propyl group, sec.-propyl, normal-butyl, sec-butyl, the tertiary butyl, allyl group, propenyl, proyl etc., and described " alkyl ", " alkenyl " and " alkynyl " can be referred to as " alkyl " or " chain alkylene ".

As be used for this paper, term " optically pure basically " is meant that optical isomer is purified basically shown in optical isomer shown in formula IIa of the present invention or the formula IIb or formula III a or the formula III b, so that its optical purity is greater than 90%, be that the shared weight percentage of optical isomer shown in optical isomer shown in formula IIa in the formula I compound or the formula IIb or formula III a or the formula III b is more than 90%, be preferably greater than 95%, be preferably greater than 98%, be preferably greater than 99%, be preferably greater than 99.5%.

The for example exemplary formula II compound of the compound of first aspect present invention can obtain according to following reaction process 1:

Flow process 1

Specifically, p-Hydroxybenzaldehyde is dissolved in the dimethyl formamide (DMF), adds salt of wormwood (K ₂CO ₃) after, generate 4-cyclopentyloxy phenyl aldehyde with the bromocyclopentane reaction.With 2, the 4-resacetophenone is dissolved in the acetone, drips the methoxychlor methane reaction behind the adding salt of wormwood, gets 2,4-two (methoxyl group methylene radical oxygen base) methyl phenyl ketone.With 2; 4-two (methoxyl group methylene radical oxygen base) methyl phenyl ketone is dissolved in the ethanol; add sodium hydroxide (NaOH); drip intermediate 4-cyclopentyloxy phenyl aldehyde; get intermediate 1-[2; 4-two (methoxy methylene radical oxygen base) benzoyl)]-2-(4 '-cyclopentyloxy phenyl) ethene; intermediate 1-[2; 4-two (methoxy methylene radical oxygen base) benzoyl)]-2-(4 '-cyclopentyloxy phenyl) ethene dissolving ethanol in, add concentrated hydrochloric acid, reflux 1-(2; 4-dihydroxy-benzene formyl radical)-2-(4 '-cyclopentyloxy phenyl) ethene; intermediate 1-(2,4-dihydroxy-benzene formyl radical)-2-(4 '-cyclopentyloxy phenyl) ethene is dissolved in the ethanol, adds sodium acetate; water; stirring and refluxing gets 4 '-cyclopentyloxy-7-hydroxyl-flavanone [that is, 2-(4-cyclopentyloxy-phenyl)-7-hydroxyl-chroman-4-on-], and molecular formula is C ₂₀H ₂₀O ₄, molecular weight is 324.

In addition, can utilize chiral column to separate synthetic 4 '-cyclopentyloxy-7-hydroxyl-flavanone raceme, obtain left-handed and two enantiomorphs of dextrorotation (formula IIa and IIb, structural formula sees above).

The various starting material that react used are that those skilled in the art can prepare according to existing knowledge, or can make by the document known method, or can buy by commerce.Used intermediate, starting material, reagent, reaction conditions etc. all can be made appropriate change according to those skilled in the art existing knowledge in the above reaction scheme.Perhaps, those skilled in the art's other formula I compound that also can fail to contain according to the second aspect present invention method.

Term used herein " composition " means and comprises the product of respectively specifying composition that comprises specified amount, and directly or indirectly from any product of the combination results of respectively specifying composition of specified amount.

Formula I chemical combination of the present invention or its isomer, raceme, enantiomorph, diastereomer, enantiomorph enriched substance, solvate and ester can also form solvate, for example hydrate, alcohol adduct etc.Above-claimed cpd can also be prodrug or the form that discharges described activeconstituents in vivo after the metabotic change.Selecting and preparing suitable prodrug derivant is technology as well known to those skilled in the art.In general, for purpose of the present invention, suitable with non-solvent compound form with the solvate form thereof of pharmacy acceptable solvent such as water, ethanol etc.

The active compound amount of gained can change the actual dose level of each activeconstituents in the pharmaceutical composition of the present invention, so that can effectively obtain required therapeutic response at concrete patient, composition and administering mode.The dosage level fibrous root is selected according to activity, route of administration, the severity of the patient's condition for the treatment of and the patient's to be treated patient's condition and the medical history of particular compound.But the way of this area is that the dosage of compound increases dosage gradually from being lower than for obtaining the level that required result of treatment requires, up to obtaining required effect.

When being used for above-mentioned treating and/or preventing or other treatment and/or when prevention, a kind of The compounds of this invention that treats and/or prevents significant quantity can be used with pure form, perhaps uses with acceptable ester of pharmacy or prodrug forms (under the situation that has these forms).Perhaps, described compound can be accepted the pharmaceutical composition administration of vehicle to contain this purpose compound and one or more medicines.The The compounds of this invention that word " treats and/or prevents significant quantity " refers to the compound of the reasonable effect that is applicable to any therapeutic treatment and/or prevention/risk than the q.s of treatment obstacle.But the total daily dosage portion that it should be understood that The compounds of this invention and composition must be examined the doctor by the master and maked decision in the medical judgment scope reliably.For any concrete patient, the concrete horizontal fibrous root of treatment effective dose is decided according to multiple factor, and described factor comprises the severity of the obstacle of being treated and this obstacle; The activity of the particular compound that is adopted; The concrete composition that is adopted; Patient's age, body weight, general health situation, sex and diet; The administration time of the particular compound that is adopted, route of administration and excretion rate; The treatment time length; The medicine that is used in combination or uses simultaneously with the particular compound that is adopted; And the known similar factor of medical field.For example, the way of this area is that the dosage of compound increases dosage gradually from being lower than for obtaining the level that required result of treatment requires, up to obtaining required effect.In general, formula I compound of the present invention is used for Mammals particularly people's dosage can be between 0.001～1000mg/kg body weight/day, for example between 0.01～100mg/kg body weight/day, for example between 0.01～10mg/kg body weight/day.

The pharmaceutical carrier that utilization those skilled in the art are familiar with can be prepared into the pharmaceutical composition of the The compounds of this invention that contains effective dosage.Therefore the present invention also provides the pharmaceutical composition that comprises with one or more nontoxic drug acceptable carriers The compounds of this invention formulated together.That described pharmaceutical composition can be mixed with especially specially is for oral administration with solid or liquid form, for the parenteral injection or for rectal administration.

Pharmaceutical composition of the present invention can by in oral, rectum, parenteral, the pond, intravaginal, intraperitoneal, part (as by powder, ointment or drops), a mouthful cheek give the mankind and other Mammalss, perhaps give as oral spray or nasal mist.Term used herein " parenteral " refers to comprise intravenously, intramuscular, intraperitoneal, breastbone is interior, subcutaneous and the administering mode of intra-articular injection and transfusion.

The composition that is suitable for the parenteral injection can comprise physiologically acceptable aseptic moisture or non-aqueous solution agent, dispersion agent, suspensoid or emulsion, and for the aseptic powder that reconstitutes sterile injectable solution agent or dispersion agent.Suitable moisture or nonaqueous carrier, thinner, solvent or vectorial example comprise water, ethanol, polyvalent alcohol (propylene glycol, polyoxyethylene glycol, glycerine etc.), vegetables oil (as sweet oil), injectable organic ester such as ethyl oleate and their suitable mixture.

These compositions also can contain auxiliary material, as sanitas, wetting agent, emulsifying agent and dispersion agent.By various antibacterial agents and anti-mycotic agent, for example parabens, trichloro-butyl alcohol, phenol, Sorbic Acid etc. can guarantee to prevent action of microorganisms.Also expectation comprises isotonic agent, for example carbohydrate, sodium-chlor etc.By using the material that can postpone absorption, for example aluminum monostearate and gelatin can reach the prolongation absorption of injectable drug form.

Remove the active ingredient beyond the region of objective existence in the suspensoid and also can contain suspension agent, for example ethoxylation i-octadecanol, polyoxyethylene sorbitol and polyoxyethylene sorbitan esters, Microcrystalline Cellulose, the mixture etc. of aluminium hydroxide, wilkinite, agar and tragacanth gum or these materials partially.

In some cases, be the effect of prolong drug, expect to slow down absorption subcutaneous or the intramuscularly medicine.This can realize by the crystal of use poorly water-soluble or the liquid suspension of amorphous substance.Like this, the absorption rate of medicine depends on its dissolution rate, and dissolution rate can be depending on crystallographic dimension and crystal formation.Perhaps, the delay of the medicament forms of parenteral admin absorb by with this medicine dissolution in or be suspended in the oily vehicle and realize.

The injectable depot formulations form can prepare by the microcapsule matrix that forms medicine in biodegradable polymer such as polylactide-poly-glycollide (polylactide-polyglycolide).Can drug releasing rate be controlled according to the character of medicine with ratio with the concrete polymkeric substance that is adopted of polymkeric substance.The example of other biological degradable polymer comprises poe class (poly (orthoesters)) and polyanhydrides (poly (anhydrides)).Injectable depot formulations also can by pharmaceutical pack is embedded in can be compatible with bodily tissue liposome or micro emulsion in prepare.

Injectable formulation can be for example by filtering with bacterial filter or sterilizing by the disinfectant that mixes the aseptic solid composite form, and described solids composition can face with before being dissolved or dispersed in sterilized water or other sterile injectable medium.

The compounds of this invention or its composition can be with oral methods or parenteral administration modes.Oral administration can be tablet, capsule, Drug coating, and the enteron aisle external application preparation has injection and suppository etc.These preparations prepare according to method appreciated by those skilled in the art.In order to make tablet, capsule, the used auxiliary material of Drug coating is the auxiliary material of conventional usefulness, for example starch, gelatin, gum arabic, silica, polyoxyethylene glycol, solvent that liquid dosage form is used such as water, ethanol, propylene glycol, vegetables oil (as Semen Maydis oil, peanut oil, olive wet goods).Contain and also have other auxiliary material in the preparation of The compounds of this invention, tensio-active agent for example, lubricant, disintegrating agent, sanitas, correctives and pigment etc.The dosage that contains formula I compound of the present invention in tablet, capsule, Drug coating, injection and suppository is that the compound amount that exists in unit dosage form is calculated.The general content of formula I compound of the present invention is 10-500mg in unit dosage form, and preferred unit dosage form contains 20-300mg.Specifically, the present invention's solid dosage for oral administration that can provide comprises capsule, tablet, pill, powder and granule.In this type of solid dosage, active compound can be accepted vehicle or carrier such as Trisodium Citrate or Lin Suanergai and/or following material with at least a inert medicine and mix: a) weighting agent or extender such as starch, lactose, sucrose, glucose, mannitol and silicic acid; B) tackiness agent such as carboxymethyl cellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and Sudan Gum-arabic; C) wetting Agent for Printing Inks such as glycerine; D) disintegrating agent such as agar, lime carbonate, potato or tapioca (flour), Lalgine, some silicate and yellow soda ash; E) solution retarding agent such as paraffin; F) absorb accelerator such as quaternary ammonium compound; G) wetting agent such as hexadecanol and Zerol; H) sorbent material such as kaolin and wilkinite and i) lubricant such as talcum powder, calcium stearate, Magnesium Stearate, solid polyethylene glycol, sodium lauryl sulphate and their mixture.Under the situation of capsule, tablet and pill, also can comprise buffer reagent in the described formulation.

The solids composition of similar type uses vehicle for example lactose and high molecular weight polyethylene glycol etc., also can be used as the weighting material in soft capsule and the hard capsule.

The solid dosage of tablet, dragee (dragees), capsule, pill and granule can prepare with dressing and shell material such as enteric coating material and known other clothing materials of field of medicine preparations.These solid dosages can be chosen wantonly and contain opalizer, and its composition also can make it just or preferentially choose wantonly with the delayed mode release of active ingredients at certain position of enteron aisle.The example of operable embedding composition comprises polymer substance and wax class.If be fit to, active compound also can be made into the micro-capsule form with one or more above-mentioned vehicle.

Liquid dosage form for oral administration comprises the acceptable emulsion of pharmacy, solution, suspensoid, syrup and elixir.Liquid dosage form also can contain this area inert diluent commonly used except that containing the active ingredient beyond the region of objective existence, for example water or other solvents, solubilizing agent and emulsifying agent be ethanol, Virahol, ethyl-carbonate, ethyl acetate, benzylalcohol, peruscabin, propylene glycol, 1,3 butylene glycol, dimethyl formamide, oils (particularly Oleum Gossypii semen, peanut oil, Semen Maydis oil, germ oil, sweet oil, Viscotrol C and sesame oil), glycerine, tetrahydrofurfuryl alcohol (tetrahydrofurfurylalcohol), polyoxyethylene glycol and the fatty acid ester of sorbitan and their mixture for example.Oral compositions also can comprise auxiliary material except that comprising inert diluent, for example wetting agent, emulsification and suspension agent, sweeting agent, correctives and flavouring agent.

The composition of confession rectum or vagina administration is suppository preferably.Suppository can be by for example theobroma oil, polyoxyethylene glycol or suppository wax are mixed and prepared with The compounds of this invention and suitable non-irritating excipient or carrier, they at room temperature are solid, therefore but next at body temperature is liquid, can melt in rectal cavity or vaginal canal and discharges active compound.

Compound of the present invention and composition thereof also consider to be used for topical.Dosage form for the topical administration The compounds of this invention comprises powder, sprays, ointment and inhalation.Under aseptic condition with active compound and pharmaceutically acceptable carrier and any required sanitas, buffer reagent or propellant mixing.Ophthalmic preparation, eye ointment, powder and solution also are considered within the scope of the present invention.

The compounds of this invention also can the liposome form administration.As known in the art, liposome makes with phosphatide or other lipid materials usually.Liposome is formed by the single or multiple lift aquation liquid crystal that is scattered in the water-bearing media.Can accept on any nontoxic, physiology that can form liposome and metabolizable lipid all can use.The present composition of liposome form also can contain stablizer, sanitas, vehicle etc. except that containing The compounds of this invention.Preferred lipid is natural and synthetic phosphatide and phosphatidylcholine (Yelkin TTS), and they can use separately or together.The method that forms liposome is well known in the art.Referring to for example Prescott, Ed., Methods in Cell Biology, Volume XIV, Academic Press, NewYork, N.Y. (1976), p.33.

In to glycyrrhizin derivatives screening active ingredients process, find that unexpectedly glycyrrhizin derivatives 4 '-cyclopentyloxy-7-hydroxyl-flavanone has tangible exciting erss hypotype (ER β) activity.The exciting ER 'beta ' activity of raceme and two enantiomorphs is compared; found that 4 '-cyclopentyloxy-7-hydroxyl-flavanone raceme and two enantiomorphs all have the activity of exciting ER β and neuroprotective; but the activity with dextrorotatory form (R type) is stronger; a little less than stimulating the breast carcinoma cell strain proliferation activity simultaneously; prompting 4 '-cyclopentyloxy-7-hydroxyl-flavanone dextrorotatory form does not have hyperplasia of tangible stimulation reproductive system or carcinogenic effect, and therefore further research all adopts dextrorotatory form to test.The invention still further relates to a kind of pharmaceutical composition that contains medicine effective dose as described compound of general formula I Ib and pharmaceutically acceptable carrier.

Discover that further 4 '-cyclopentyloxy-7-hydroxyl-flavanone dextrorotatory form has significant vasorelaxation action, and neurone is also had obvious protection and nutritional activities, effect is better than Liquiritigenin.4 '-cyclopentyloxy-7-hydroxyl-flavanone dextrorotatory form has promoter action and strengthens the voluntary movement ability of Parkinson disease model mouse the Model of Dementia learning and memory of little mouse.In addition, this class glycyrrhizin derivatives has obvious anti-ulcerative colitis effect, reduces the level of inflammation-related factor tumor necrosis factor alpha and interleukin 1 β, to the therapeutic action that the rat kind rheumatic arthritis has, reduces the level of tumor necrosis factor alpha.Activity in vivo obviously is better than Liquiritigenin.

Compound shown in the formula I of the present invention is the new compound that a class has the Liquiritigenin parent nucleus, and they can be used for prevention and treatment ischemic cerebrovascular disease, nerve degenerative diseases, and treat ulcerative colitis, rheumatoid arthritis.

In addition, according to result of study of the present invention as seen, compound shown in the formula I of the present invention can be used as with the next item down or multinomial drug use: the i) agonist of erss hypotype (ER β); Ii) hypermnesia agent; Iii) neuroprotective; Iv) analgesic agent.

Compound shown in the formula I of the present invention has the long transformation period, the transformation period of Liquiritigenin is 141.2 minutes (min), and the transformation period with strong active 4 '-cyclopentyloxy-7-hydroxyl-flavanone dextrorotatory form reaches 424.8min, transformation period obviously prolongs, thereby can the administration every day 1-2 time, and Liquiritigenin is short because of the transformation period, needs administration 4-5 time every day, becomes the property of medicine relatively poor.On the other hand, compare with Liquiritigenin, The compounds of this invention has increased lipotropy after introducing and encircling amyl ether, helps intestinal absorption, increases bioavailability, and is stored in fatty tissue and the cerebral tissue, slowly discharges and a long-acting later on.

Compound shown in the formula I of the present invention can be used for the treatment of and/or prevent disease of the present invention effectively.

Description of drawings

Fig. 1 is the color atlas of 4 '-cyclopentyloxy-7-hydroxyl-flavanone raceme chiral separation.

Fig. 2 is one of 4 '-cyclopentyloxy-7-hydroxyl-flavanone enantiomorph (dextrorotatory form) mass spectrometric detection figure.

Embodiment

Further specify the present invention below by specific embodiment, still, should be understood to, these embodiment are only used for the more detailed usefulness that specifically describes, and are used for limiting in any form the present invention and should not be construed as.

The present invention carries out generality and/or concrete description to the material and the test method that are used in the test.Though for realizing that employed many materials of the object of the invention and working method are well known in the art, the present invention still does to describe in detail as far as possible at this.It will be apparent to those skilled in the art that hereinafter, if do not specify that material therefor of the present invention and working method are well known in the art.Main determining instrument used herein is: molten point is Zabspec type velocitron, MadLab Acquisition and Processing of Biomedical Signal, flow cytometer (Beckman Coulte), CS-2 type mouse autonomic activities program instrument, rat foot cubic content measurement instrument (Japanese Shimadzu Corporation) with XY-1 type electric heating melting point instrument, mass spectrograph.Laboratory animal is all available from Beijing Vital River Experimental Animals Technology Co., Ltd..Liquiritigenin is available from Nat'l Pharmaceutical ﹠ Biological Products Control Institute standard substance chamber.Except that special indicating, the chemical reagent of employing is all available from Beijing chemical reagents corporation.

The preparation of embodiment 1:4-cyclopentyloxy phenyl aldehyde

P-Hydroxybenzaldehyde 12.2 grams (0.1mol) are dissolved among 100 milliliters of DMF, add K ₂CO ₃13.8 gram (0.1mol) stirs half an hour, disposable adding bromocyclopentane is heated to 100 ℃ of reactions 5 hours, cold filtration, and DMF, residuum CH are removed in decompression ₂Cl ₂Anhydrous MgSO is washed in dissolving 3 times ₄Dry removal of solvent under reduced pressure gets product 4-cyclopentyloxy phenyl aldehyde 17.2 grams, yield 90.5%.

Embodiment 2:2, the preparation of 4-two (methoxyl group methylene radical oxygen base) methyl phenyl ketone

With 2,4-resacetophenone 15.2 grams (0.1mol) are dissolved in 200 milliliters of acetone, add under salt of wormwood 27.6g (0.2mol) room temperature and stir half an hour, drip methoxychlor methane 24 grams (0.3mol), dropwise in half an hour, stir under the room temperature and spend the night, acetone is removed in decompression, and residuum is dissolved in water, dichloromethane extraction 100 milliliters * 3 times, merge organic phase, wash anhydrous MgSO 2 times ₄Dry removal of solvent under reduced pressure gets colourless liquid 25 grams, yield 98.2%.

Embodiment 3:1-[2,4-two (methoxy methylene radical oxygen base) benzoyl)]-2-(4 '-ring penta Oxygen base phenyl) preparation of ethene

With intermediate 2,4-two (methoxyl group methylene radical oxygen base) methyl phenyl ketone 12 grams (0.05mol) are dissolved in 150 milliliters of ethanol, add 8 gram NaOH (0.2mol), drip synthetic 4-cyclopentyloxy phenyl aldehyde 10.5 grams (0.06mol) among the embodiment 1, finish and refluxed removal of solvent under reduced pressure 2 hours, residue dissolves with methylene dichloride, wash 3 times drying, removal of solvent under reduced pressure gets product 16.8 grams, yield 84.1%.

Embodiment 4:1-(2,4-dihydroxy-benzene formyl radical)-2-(4 '-cyclopentyloxy phenyl) ethene Preparation

Product 16.1 grams (0.4mol) that embodiment 3 is obtained are dissolved in 100 milliliters of ethanol, add 10 milliliters of concentrated hydrochloric acids, reflux and stir 2 hours, and ethanol is removed in decompression, and residuum dissolves with methylene dichloride, washing, anhydrous MgSO ₄The organic phase methylene dichloride is removed in dry decompression, sherwood oil grinds to such an extent that white solid product 1-(2,4-dihydroxy-benzene formyl radical)-2-(4 '-cyclopentyloxy phenyl) ethene 10.8 restrains yield 84.5%.

The preparation of embodiment 5:4 '-cyclopentyloxy-7-hydroxyl-flavanone raceme

With intermediate 1-(2; 4-dihydroxy-benzene formyl radical)-2-(4 '-cyclopentyloxy phenyl) ethene 6.3 gram (0.02mol) is dissolved in 100 milliliters of ethanol, adds sodium acetate 40 grams, 50 milliliters in water; stirring and refluxing cooling in 10 hours; ethanol is removed in decompression, the water dichloromethane extraction, and drying has machine; filter; remove methylene dichloride, residuum grinds to such an extent that white solid 3.2 restrains yield 50.3% with sherwood oil.

¹H?NMR(δ)ppm(DMSO-d6)：1.68(6H，m)，1.91(2H，m)，2.65(1Hdd，J＝18Hz)，3.13(1H，m)，4.83(1H，m)，5.48(1H，m)，6.34(1H，d，J＝2.0Hz)，6.51(1H，dd，J＝8Hz)，6.92(2H，d，J＝8.0Hz)，7.42(2H，d，J＝8Hz)，7.65(1H，d，J＝6.4Hz)，10.57(1H，S)，HCL-001T，MS(FAB)：325(M+1)。

The chiral separation and the evaluation of embodiment 6:4 '-cyclopentyloxy-7-hydroxyl-flavanone

Adopt the C18 post to separate embodiment 5 synthetic 4 '-cyclopentyloxy-7-hydroxyl-flavanone raceme.Chromatographic column is chiral column CJ-H, diameter 5 μ m, and moving phase is normal hexane: ethanol=85: 15, detector are PDA.Obtain the chirality enantiomorph by separation, the retention time at former and later two peaks is respectively 7min and 12min (seeing Figure of description 1).Mass spectrometry results shows that the molecular weight of raceme and two enantiomorphs is 324 (seeing Figure of description 2).

Detect the specific rotation that separation obtains two enantiomorphs with AUTOPOL III type polarimeter.Polarization tube is 1 decimeter light path, wavelength 589nm, and temperature 25 degree, solvent is a water.Leading peak is dextrorotatory form (a R type) as a result, and specific rotation is+0.062, and postpeak is levo form (a S type), and specific rotation is-0.062, specific rotatory power (water).The optical purity of the product of two enantiomorphs that obtain is 95%.

The preparation of embodiment 7:4 '-cyclopentyl methanoyl-7-hydroxyl-flavanone

Liquiritigenin 12.8 grams (0.05mol) are dissolved in the 100mL chloroform, add pyridine 10mL, frozen water is cooled to about 5 ℃, drips cyclopentyl formyl chloride 13.2 grams (0.1mol), stirring at room 5 hours, reaction mixture washes with water 3 times, anhydrous magnesium sulfate drying filters, and concentrates, concentrated solution silica gel column chromatography (sherwood oil: ethyl acetate=3: 1) get target compound 7.5 grams, yield 42.3%.Products therefrom can further carry out chiral separation.

¹H?NMR(δ)ppm(DMSO-d6)：1.58(6H，m)，1.81(2H，m)，2.35(1Hdd，J＝18Hz)，3.13(1H，m)，4.73(1H，m)，5.40(1H，m)，6.24(1H，d，J＝2.0Hz)，6.41(1H，dd，J＝8Hz)，6.72(2H，d，J＝8.0Hz)，7.32(2H，d，J＝8Hz)，7.65(1H，d，J＝6.4Hz)，10.67(1H，S)，MS(FAB)：353(M+1)。

Embodiment 8:4 '-cyclopentyloxy-7-hydroxyl-flavanone chipal compounds excitement The ER 'beta ' activity relatively

With reference to the previous medicaments sifting model of setting up (Acta Pharmaceutica Sinica, 2006,41 (8): 721-726), detect ER β hypotype agonist activity based on reporter gene and erss hypotype (estrogenreceptor β, ER β) signal path.

The cell strain of stable transfection ER β expression vector and reporter gene carrier is cultivated with the foetal calf serum (available from Hyclone company) that contains 5% no female activity, 1640 no phenol red nutrient solutions.Cell is after trysinization, with 1 * 10 ⁵/ ml cell count is spread 96 orifice plates, 180 μ l/ holes, add each the concentration medicine 20 μ l/ hole that has prepared, act on after 72 hours, detect luminous value, with the luminous value of sample well luminous value divided by the nutrient solution control wells, obtain the abduction delivering multiple, with the activity of the exciting ER β of reflection sample, multiple is high more, and the activity of exciting ER β is strong more.

Found that sample 4 '-ring amyl ether-7-hydroxyl-flavanone raceme and two enantiomorph all can be induced the beta mediated reporter gene expression of ER, but with dextrorotatory form induce multiple higher, apparently higher than Liquiritigenin (table 1).The result shows that 4 '-ring amyl ether-7-hydroxyl-flavanone dextrorotatory form (R type) is strong ER beta-agonists.

Table 1: sample 4 '-ring amyl ether-7-hydroxyl-flavanone and enantiomorph thereof are to the influence of the beta mediated reporter gene expression of ER

The transformation period is detected in the body of embodiment 9:4 '-cyclopentyloxy-7-hydroxyl-flavanone

The Wistar rat oral gavage gives 4 '-cyclopentyloxy-7-hydroxyl-flavanone (dextrorotatory form) or Liquiritigenin, and dosage is 60mg/kg, prepares with 1% Xylo-Mucine methanol mixed solution.Behind medicine 5,30,60,120,180,300,480,660,1200min gets blood 0.5ml from eye socket, place the pipe of heparin, it is centrifugal that (3500rmp 15min), gets supernatant 0.2ml, mark working fluid (icarin in accurate the absorption, 0.2mg/ml) 2 μ L mixings, add methyl alcohol 0.8mL and on vortex mixer, mix the centrifugal 5min of 10000r/min, draw supernatant liquid, volatilize under the nitrogen gas stream.The moving phase dissolving detects.Waters 2695 high performance liquid chromatographs (HPLC), Waters 2487UV detector, Empower Personal chromatographic working station.Chromatographic column: Xterra C18 post 5 μ m (4.6 * 250mm).Moving phase is methyl alcohol: acetonitrile: water (containing 0.2%TFA)=20: 50: 30.After the methodology checking and setting up typical curve, adopt area normalization method, calculate the Plasma Concentration of each time point sample, with statistics apart from the bioavailability of method calculation sample and elimination transformation period mutually, calculation result shows, the absolute bioavailability of 4 '-cyclopentyloxy-7-hydroxyl-flavanone dextrorotatory form (R type) is 70%, transformation period (t _1/2) be 424.8min, the absolute bioavailability of Liquiritigenin is 30%, and the transformation period is 141.2min, and the result shows that the bioavailability of 4 '-cyclopentyloxy-7-hydroxyl-flavanone dextrorotatory form (R type) obviously improves, and the transformation period obviously prolongs.

Embodiment 10:4 '-cyclopentyloxy-7-hydroxyl-flavanone is to the brain basal arteries Dilating effect

Get the brain basal arteries of anesthetized dog, separate clean circumvascular reticular tissue, be cut into the vascular circle that is about 0.6cm, lateral suspension is on two trilateral stainless steel frames, place the 10ml double-layer heat insulation bath that fills Krebs liquid, bath temperature is (37 ± 0.5) ℃, and is not open close to contain 5%CO ₂Mixture of oxygen one side tripod be fixed in the ventilation hook on, opposite side is connected on the JZ101 type muscular tension transmitter, tension variation writes down and handles with the MadLab Acquisition and Processing of Biomedical Signal.Vascular circle adds the 0.5g rest tension and carries out balance, and every 20min changes liquid 1 time in the balance period, experimentizes behind the 2h.With final concentration is that the Repone K (KCl) of 20mmol/L causes that arterial ring shrinks, waiting to shrink the steady back of curve accumulation adding final concentration is 1,5,25,125,4 '-cyclopentyloxy of 625 μ g/ml-7-hydroxyl-flavanone dextrorotatory form (0.1% dimethyl sulfoxide (DMSO) DMSO dissolving), record shrinks height (mm) decline scope, with the contraction before the dosing highly is 100, calculate the percentage ratio that the dosing post shrinkage highly descends, per twice dosing be spaced apart 5min (each dosing after in a basic balance), each arterial ring is only traced a curve.5 samples of each concentration represent that with mean number ± standard deviation the result is as shown in table 2.The result shows that 4 '-ring brain basal arteries that amyl ether-7-hydroxyl-flavanone dextrorotation physical efficiency antagonism KCl causes shrink, and improve the cerebral blood supply state.

Table 2: sample 4 '-ring amyl ether-7-hydroxyl-flavanone causes the influence that dog brain basal arteries shrink to KCl

Annotate: compare with the DMSO+KCl group, ^*P＜0.05, ^{* *}P＜0.01, ^{* *}P＜0.001

Embodiment 11: sample 4 '-ring amyl ether-7-hydroxyl-flavanone is focal to rat acute The influence of property cerebral ischemia

The Wistar rat, male, body weight 210 ± 20 grams.Animal is divided at random by body weight: normal control group, model control group, sample 4 '-ring amyl ether-7-hydroxyl-flavanone dextrorotatory form 10mg/kg group, 20mg/kg group, 30mg/kg group and Liquiritigenin 30mg/kg group, 10 every group.Irritate stomach by body weight and give each soup 0.5ml/100g body weight, normal control group and model control group give equal-volume physiological saline water, and all successive administration is 3 days.1h behind the 2nd day medicine, vetanarcol (35mg/kg) intraperitoneal injection of anesthesia.Press literature method, expose rat right side arteria cerebri media, have the small pieces quantitative paper of 50% liquor ferri trichloridi to spread on the arteria cerebri media suction,, sew up wound with taking off filter paper behind the dissecting microscope observation thrombosis.Control group covers the iron trichloride filter paper the same model group of all the other operating procedures except that not applying.23h gastric infusion 1 time again after the modeling.24h after modeling, broken end is got brain.The mensuration of cerebral infarct size: rat broken end back is taken out full brain rapidly, removes olfactory bulb, cerebellum and low brain stem, and remainder is crown under 4 ℃ to be cut into 5.(every 5ml dye liquor contains 4%TTC 1.5ml, 1mol/L K rapidly the brain sheet to be placed chloro triphenyltetrazolium chloride (TTC) dye liquor ₂HPO ₄0.1ml), 37 ℃ of lucifuge temperature are incubated 30min, take out in rearmounted 10% formaldehyde solution to keep in Dark Place.The non-ischemic region in dyed back is a rose, and infarct is a white.The white tissue is weighed careful the separation, account for the per-cent of total brain weight as cerebral infarct size with blocking tissue's weight.The mensuration of brain water content: rat broken end back is taken out full brain rapidly, gets right half brain ball, claims weight in wet base, and 110 ℃ of oven dry 24h claim dry weight, calculate brain water content.

Found that sample 4 '-ring amyl ether-7-hydroxyl-flavanone dextrorotation physical efficiency is obviously dwindled brain infarction area, reduce the water content of arteria cerebri media embolism rat brain, effect is better than the Liquiritigenin (seeing Table 3) with dosage.

Table 3:4 '-ring amyl ether-7-hydroxyl-flavanone and Liquiritigenin are to the influence of rat acute focal cerebral ischemia

Annotate: compare #P＜0.05 with the normal control group; Compare with model control group, ^*P＜0.05, ^*P＜0.01

Embodiment 12:4 '-cyclopentyloxy-7-hydroxyl-flavanone anti-beta amyloid Cause the effect of rat Neuron Apoptosis

Adopt Flow cytometry 4 '-cyclopentyloxy-7-hydroxyl-flavanone raceme and isolating enantiomorph to anti-beta amyloid (A β _25～35, Sigma company) and cause the anti-apoptotic effect of rat neurocyte of former generation.SD rat suckling mouse, in the birth 12h, ♀

Dual-purpose.The rat suckling mouse is after volume fraction is 75% ethanol disinfection, and broken end places and contains the plate of dissecting liquid, isolates full brain, and dissecting microscope separates hippocampal tissue down.Be placed in 0.25% trypsinase and and shred with scissors.Draw trypsinase to containing in the serum centrifuge tube every 4min, and add fresh trypsinase until no macroscopic tissue block.200 order metallic sieves filter the back with the centrifugal 10min of 1000rpm.Its DMEM/F-12 kind that is resuspended in 10% foetal calf serum is planted liquid, be inoculated in 24 orifice plates that wrap in advance by poly-lysine, be changed to the DMEM/F-12 nutrient solution of 3% foetal calf serum, 1%N2,1%B27 and 3ng/mL cytosine arabinoside after the overnight incubation.Experiment is grouped as follows: normal control group, A β _25～35The transaction module group; A β _25～35+ different concns sample sets, sample are respectively 4 '-cyclopentyloxy-7-hydroxyl-flavanone raceme or isolating two enantiomorph final concentrations are 0.2,1,5 μ g/mL, A β _25～35Concentration be 15 μ mol/L.Detect apoptosis after cultivating 24h.The cell suspension of getting 100 μ l adds 5 μ l Annexin V-FITC and 10 μ l PI solution (20 μ g/ml) mixings in the flow cytometer detector tube, the room temperature lucifuge was hatched 15 minutes, added 400 μ l PBS, flow cytometry analysis.

The result is as shown in the table, and with normal control group ratio, the apoptosis rate of A β transaction module group obviously increases, and statistical significance (P＜0.01) is arranged; Compare with A β transaction module group, the apoptosis rate that respectively adds sample sets significantly reduces (P＜0.01), and with the increase of culture system Chinese traditional medicine concentration, apoptosis rate reduces more obvious, the apoptosis rate of raceme and two enantiomorphs all is lower than Liquiritigenin under each concentration, but comparatively speaking, the apoptosis rate lower (table 4) of 4 '-cyclopentyloxy-7-hydroxyl-flavanone dextrorotatory form.

The result shows; 4 '-cyclopentyloxy-7-hydroxyl-flavanone raceme and two enantiomorph all have anti-A β to cause the activity of former generation Neuron Apoptosis; play neuroprotective, effect is better than Liquiritigenin, but the anti-Neuron Apoptosis activity of dextrorotatory form is better than raceme or its levo form.

Table 4: the anti-A β of sample _25～35Cause the effect of former generation Neuron Apoptosis

Annotate: compare ##P＜0.01 with the normal control group; With the model group ratio, ^*P＜0.01, ^*P＜0.01, ^{* *}P＜0.001; Compare ， @P＜0.05 with Liquiritigenin

With with 4 ' of dosage-ring amyl ether-7-hydroxyl-flavanone (S type) ， $P＜0.05 relatively

Embodiment 13:4 '-cyclopentyloxy-7-hydroxyl-flavanone is to the Model of Dementia mouse The influence of learning and memory

Adopt mouse step down test and Morris water maze test, observation 4 '-cyclopentyloxy-7-hydroxyl-flavanone dextrorotatory form is to the influence of Model of Dementia learning and memory of little mouse due to the Scopolamine.Kunming mice, body weight 14～16g, female.Mouse is by body weight isostatic principle randomly assigne grouping administration (dosage sees the following form).Behind the administration 7d, (5mg/kg, Sigma) modeling and carry out following step down test: animal is placed on the platform, and the 3min that conforms passes to 36V voltage to the abdominal injection scopolamine hydrobromide then, training 5min.Test behind the 24h, record jumps off for the first time the latent period of platform and the errors number in the 5min, with this as the memory achievement.

The Morris water maze test is as follows: test and earlier mouse was put into water maze free swimming 2min to conform in preceding 1 day.15min abdominal injection Scopolamine 5mg/kg before the morning every day training gives the sample 4 '-cyclopentyloxy-7-hydroxyl-flavanone of various dose or Liquiritigenin once (dosage sees the following form) every day.First day mouse training 3 times is put into water with mouse towards wall of container from 3 different place of entry respectively at every turn and is swum, if mouse can not be found underwater platform in 90s, then directly places it on platform and stops 30s.Each training second day morning and afternoon once.Write down the 3rd day morning and afternoon and climb up after the mouse entry the used time of platform (latent period), with the latent period of morning and afternoon average, calculate mean value in latent period of each group.

The step down test result shows that with normal control group ratio, errors number obviously increases in the model group 5min, latent period (second, s) significantly shorten.With the model group ratio, 4 '-cyclopentyloxy-7-hydroxyl-flavanone group dextrorotatory form or Liquiritigenin group errors number obviously reduce, and obvious prolongation (seeing Table 5) is also arranged latent period, show that the learning and memory of model mice after the administration strengthens.Compare with the Liquiritigenin with dosage, the effect of 4 '-cyclopentyloxy-7-hydroxyl-flavanone dextrorotatory form is better.

Morris water maze result shows that compare with the normal control group, obviously prolong the latent period of model group (Scopolamine 5mg/kg) mouse.Obviously shorten the latent period of 4 '-cyclopentyloxy-7-hydroxyl-flavanone group dextrorotatory form or Liquiritigenin group, is dose-dependence (table 6), shows that the learning and memory of model mice after the administration improves.Liquiritigenin shortens preclinical effect and is weaker than with 4 ' of dosage-cyclopentyloxy-7-hydroxyl-flavanone dextrorotatory form group.

Table 5:4 '-cyclopentyloxy-7-hydroxyl-flavanone or Liquiritigenin are to the influence of model mice diving tower achievement

Annotate: compare with the normal control group: ###P＜0.001; Compare with model group: ^*＜0.05, ^*P＜0.01, ^{* *}P＜0.001; Compare ， @P＜0.05 with Liquiritigenin

Table 6:4 '-cyclopentyloxy-7-hydroxyl-flavanone or Liquiritigenin are to the preclinical influence of appearing on the stage of model mice water maze

Annotate: compare with the normal control group: ##P＜0.01; Compare with model group: ^*P＜0.05, ^*P＜0.01; Compare ， @P＜0.05 with Liquiritigenin

Embodiment 14:4 '-cyclopentyloxy-7-hydroxyl-flavanone causes the handkerchief gold to MPTP The influence of gloomy disease model spontaneous activity in mice

The C57BL/6J mouse, male and female half and half, body weight 20 ± 1 grams.1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine, MPTP, Sigma company).With the mouse random packet, 10 every group, (dosage is 10,2030mg/kg) or Liquiritigenin (30mg/kg) to be respectively normal control group, model group and 4 '-cyclopentyloxy-7-hydroxyl-flavanone dextrorotatory form group.Continuous gastric infusion 14d, in the 11st day the beginning gastric infusion before 1h abdominal injection MPTP 30mg/kg (control group gives isopyknic physiological saline), continuous 4d, 1d after last 1 administration carries out the study of behaviour index test, and mouse is put into the autonomic activities case, measure 4 mouse simultaneously at every turn, in each active box 1, write down the mouse activity automatically by registering instrument, measure the movable number of times in every mouse 5min.

Except that the normal control group, all the other are respectively organized mouse and in various degree dyskinesia all occurs, show as slow movement, tremble, perpendicular hair, perpendicular tail and movable reduce etc.Compare with model group, 4 '-cyclopentyloxy-7-hydroxyl-autonomic activities the number of times of flavanone dextrorotatory form group mouse in the test duration significantly increases, be dose-dependence, prompting is subjected to test product can improve the central nervous system excitement degree and the voluntary movement ability of model mice, PD there is therapeutic action, the Liquiritigenin group also has positive effect, and effect is weaker than with 4 ' of dosage-cyclopentyloxy-7-hydroxyl-flavanone dextrorotatory form group (the results are shown in Table 7).

Table 7:4 '-cyclopentyloxy-7-hydroxyl-flavanone causes the influence of Parkinson disease model spontaneous activity in mice to MPTP

Annotate: compare with normal group: ###P＜0.001; Compare with model group: ^*P＜0.05, ^*P＜0.01; Compare ， @P＜0.05 with Liquiritigenin

Embodiment 15: glycyrrhizin derivatives is to the therapeutic action of rat ulcer colitis

The SD rat is divided into normal control group, model group and 4 '-cyclopentyloxy-7-hydroxyl-flavanone dextrorotatory form low dose group, middle dosage group, high dose group, 4 '-ring penta methanoyl-7-hydroxyl-flavanone high dose group and Liquiritigenin high dose group at random, and concrete dosage sees the following form.The normal control group is not done any processing, model group is with trinitro-benzene-sulfonic acid (TNBS, Sigma company) bowel lavage, glycyrrhizin derivatives 4 '-cyclopentyloxy-7-hydroxyl-flavanone group, 4 '-ring penta methanoyl-7-hydroxyl-flavanone group and Liquiritigenin group are respectively with gastric infusion after the trinitro-benzene-sulfonic acid modeling.SD rat fasting 24h freely drinks water during modeling, etherization, and the silicone tube that is about 12cm with a diameter 2.0cm is by the about deeply 8cm of the light and slow insertion of anus, and 50% ethanolic soln that will contain the TNBS of 150mg/kg slowly pushes colon, makes the animal nature that keeps lying low clear-headed.Control animals gavages physiological saline 2ml/ time after the modeling, 4 '-cyclopentyloxy-7-hydroxyl-flavanone dosage is respectively 10,20,30mg/kg, the dosage of 4 '-ring, penta methanoyl-7-hydroxyl-flavanone and Liquiritigenin is 30mg/kg, gastric infusion, every day 1 time.Dead 2 rats in 3 week every component other places, back are got the colon sample and do the pathology inspection, to determine the modeling success.All experimental rats are disposed in the anesthesia back, and the abdominal cavity animal is got 2 milliliters of blood, separation of serum, and-20 ℃ of preservations, (test kit is available from R﹠amp for level to use ELISA method mensuration tumor necrosis factor alpha (TNF-α) and interleukin-11 β (IL-1 β) by the test kit requirement; D company).Get anus to caecum intestinal tube (about 8cm), cut off, rinse a back part well with cold saline and fix, carry out the scoring of naked eyes general form and histology form with 4.0% Paraformaldehyde 96 along the mesentery longitudinal axis.With the sample specimens paraffin embedding slices, HE dyes, assess inflammation and ulcer under colon's mirror after the classification.General form damage scoring index comprises adhesion, contrafluxion, ulcer and inflammation.Adhesion and congested by the weight that has that it's too late 0,1,2 minute respectively, every increases of scope 1cm scores and all adds 1 when inflammation, ulcer number occurring and increasing by 1, ulcer area＞2cm.Histological indices comprises ulcer, inflammation, granuloma, fibrosis and the pathology degree of depth, and by the weight that has that it's too late 0,1,2 minute respectively, the pathology degree of depth reaches submucosa, flesh layer, placenta percreta to be counted respectively 1,2,3 fen, and every addition gets total points.

The general form observations shows that normal control group general form is normal, no oedema, hemorrhage, ulcer.The visible intestinal mucosa congestion and edema of TNBS model group, necrosis, ulceration.4 '-cyclopentyloxy-7-hydroxyl-above-mentioned variation of flavanone dextrorotatory form treated animal all obviously alleviates than model group, and increases with dosage, and above-mentioned pathology alleviates more obvious.4 '-ring, penta methanoyl-7-hydroxyl-flavanone group and Liquiritigenin group are also effective, wherein a little less than the Liquiritigenin group effect relatively (table 8).

The histological examination result shows, visible normal control group inorganization damage performance under the mirror of HE dyeing back.The visible mucous membrane of TNBS model group, submucosa even flesh layer massive inflammatory cells infiltrated mainly show as neutrophil infiltration, monocyte and eosinophilic granulocyte as seen, as seen hemorrhage, the oedema of submucosa, ulcer and mucomembranous gland occurs and arrange distortion.4 '-cyclopentyloxy-visible mucous hyperemia oedema of 7-hydroxyl-flavanone senior middle school dosage treated animal alleviates, ulcer healing, and glandular hyperplasia is obvious, and the colon structure is normal substantially.4 '-ring, penta methanoyl-7-hydroxyl-flavanone group and Liquiritigenin group are also effective, and wherein the effect of Liquiritigenin is weaker than with 4 ' of dosage-cyclopentyloxy-7-hydroxyl-flavanone dextrorotatory form (table 8).

Detect inflammatory mediator result demonstration in the serum, TNF-α and IL-1 β level are apparently higher than normal control group (P＜0.001) in the model group animal serum.4 '-cyclopentyloxy-7-hydroxyl-TNF-α of flavanone dextrorotatory form administration group and the content of IL-1 β are lower than model group, increase with dosage, descend by obvious more (table 9).

Table 8: glycyrrhizin derivatives is to the influence of ulcerative colitis rat model colonic pathological change

Annotate: compare with normal group: ###P＜0.001; Compare with model group: ^*P＜0.05, ^*P＜0.01, ^{* *}P＜0.001; Compare ， @P＜0.05 with Liquiritigenin

Table 9: glycyrrhizin derivatives is to the influence of ulcerative colitis rat model inflammatory factor

Embodiment 16:4 '-cyclopentyloxy-7-hydroxyl-flavanone causes mouse acetic acid and turns round The influence of precursor reactant

Get 40 of Kunming kind small white mouses and divide 4 groups at random, male and female half and half, gavage administration, lh after administration, abdominal injection l% acetic acid physiological saline (Glacial acetic acid: 0.0lml/g body weight physiological saline 1: 100), observe respectively organize mouse in the 30min turn round the body number of times, experiment is divided into physiological saline control group, 4 '-cyclopentyloxy-7-hydroxyl-flavanone dextrorotatory form group (dosage be 10,20,30mg/kg) and Liquiritigenin group (30mg/kg).

Found that, after mouse gives sample 4 '-cyclopentyloxy-7-hydroxyl-flavanone dextrorotatory form, the 30min writhing response that causes for the acetic acid chemical factor obviously reduces, relatively difference is obvious with the physiological saline group, dosage is big more, effect is obvious more, shows that 4 '-cyclopentyloxy-7-hydroxyl-flavanone dextrorotatory form has analgesic activity, and effect is better than the Liquiritigenin group (table 10) with dosage.

Table 10:4 '-cyclopentyloxy-7-hydroxyl-flavanone causes the influence of writhing response to mouse acetic acid

Annotate: compare with the physiological saline group: ^*P＜0.01, ^{* *}P＜0.001; Compare ， @P＜0.05 with Liquiritigenin

Embodiment 17:4 '-cyclopentyloxy-7-hydroxyl-flavanone closes rat kind rheumatism The therapeutic action that joint is scorching

The SD male rat, body weight 200 ± 6 grams.Get 1 (10m1) (Sigma company) of Freund's complete adjuvant, under the aseptic condition, every right back sufficient sole of the foot intracutaneous of modeling rat is annotated people 0.1m1 adjuvant and is caused inflammation, duplicates rheumatoid arthritis (RA) rat model.Only injecting normal saline 0.lml/ of normal group (5).Sufficient volume about measuring before the modeling is got the average foundation of dividing into groups before as medicine.Except that normal control group and model control group, modeling begins back administration treated animal and irritates stomach every day and give 4 '-cyclopentyloxy-7-hydroxyl-flavanone dextrorotatory form (dosage be respectively 10,20,30mg/kg) and Liquiritigenin (30mg/kg) 1 time, continuous 30 days.Respectively at after the modeling 2 days, 7 days, 14 days, 21 days and 30 days, survey right sufficient volume once.Paw swelling method of calculation: the scorching front foot volume of swelling degree (%)=(cause scorching metapedes volume-cause scorching front foot volume)/cause.When experiment finished, anesthetized animal was got 2 milliliters of abdominal vein blood, and separation of serum is with ELISA test kit (R﹠amp; D company) the by specification method is surveyed blood serum tumor necrosin ﹠ (TNF-α) level.

The result shows that rat is behind the injection Freund's complete adjuvant, and model control group injection foot (right foot) tangible swelling occurred in the 2nd day, and sufficient volume is obviously greater than the normal control group.The rat paw edema degree of administration group is starkly lower than model control group, and increases with dosage, and the rat paw edema degree descends by more obvious (table 11), and TNF-α also is starkly lower than model control group (table 12) in the group of the administration simultaneously serum.The result shows, 4 '-cyclopentyloxy-7-hydroxyl-flavanone dextrorotatory form has the arthroncus of significant inhibition RA rat, prompting has therapeutic action to rheumatoid arthritis, and the effect of Liquiritigenin is weaker than with 4 ' of dosage-cyclopentyloxy-7-hydroxyl-flavanone dextrorotatory form.

Table 11:4 '-cyclopentyloxy-7-hydroxyl-flavanone dextrorotatory form is to the therapeutic action of rat RA

Annotate: compare with model control group: ^*P＜0.01, ^{* *}P＜0.001; Compare ， @P＜0.05 with Liquiritigenin

Table 12:4 '-cyclopentyloxy-7-hydroxyl-flavanone dextrorotatory form is to the influence of RA rat blood serum TNF-alpha levels

Annotate: compare with normal group: ##P＜0.01; Compare with model group: ^*P＜0.05, ^*P＜0.01; Compare ， @P＜0.05 with Liquiritigenin

Embodiment 18:4 '-cyclopentyloxy-7-hydroxyl-flavanone is to the MCF-7 cell The influence of propagation

Human breast carcinoma MCF-7 cell is gone down to posterity 2 times with the no phenol red RPMI-1640 that contains 10% no female serum, digest according to a conventional method centrifugal, with 1.2 * 10 ⁴The concentration in/180 μ l/ holes is inoculated in 96 orifice plates, treat that the adherent complete back of 18-24 hour cell adds 4 '-cyclopentyloxy-7-hydroxyl-flavanone raceme, two enantiomorphs, Liquiritigenin or estradiol 20 μ l/ holes of proper concn, continue to cultivate 72 hours, before off-test, add MTT (5mg/ml) 20 μ l/ holes, cultivate supernatant discarded after 4 hours, add DMSO 180 μ l/ holes, 540nm place, concussion dissolving back measures the OD value, calculates cell proliferation rate.The results are shown in Table 13.

Found that, Liquiritigenin has the effect of stimulating cellular proliferation when concentration 50 μ g/ml, but along with concentration reduces, promote the active of cell proliferation obviously to descend, the raceme of 4 '-cyclopentyloxy-7-hydroxyl-flavanone and the activity that levo form stimulates cellular proliferation are similar to the effect of Liquiritigenin, and the promotion cell-proliferation activity of 4 '-cyclopentyloxy-7-hydroxyl-flavanone dextrorotatory form relatively weak (table 13), show that 4 '-cyclopentyloxy-7-hydroxyl-flavanone dextrorotatory form does not have tangible female activity, point out its potential side effect such as the non-stimulated mammary gland of life-time service or endometrial epithelial cell propagation or hyperplasia clinically.

Table 13:4 '-cyclopentyloxy-7-hydroxyl-flavanone is to the influence of MCF-7 cell proliferation

Annotate: compare with Liquiritigenin, 4 '-cyclopentyloxy-7-hydroxyl-flavanone raceme and its levo form group with dosage, ^*P＜0.01, ^*P＜0.05.

Claims

1, formula I compound:

Wherein:

N is 0 or 1;

R ₁Be C _3-8Cycloalkyl;

R ₂Be H,

2, the compound of claim 1:

Wherein:

N is 0;

R ₁Be selected from: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, suberyl;

R ₂Be H,

3, the compound of claim 1:

Wherein:

N is 1;

R ₂Be H,

4, the compound of claim 1, it is selected from:

5, the compound of claim 1, it is selected from:

And enantiomorph enriched substance, solvate, prodrug and ester.

6, the compound of claim 1, it is:

Basically optically pure

Basically optically pure

Basically optically pure

Or

Basically optically pure

And solvate.

7, the method for preparing the described compound of claim 1, this method may further comprise the steps:

A) make p-Hydroxybenzaldehyde and formula

Shown in compound reaction, obtain formula

Compound;

B) make formula

Compound and formula Hal-CH ₂OCH ₃The compound reaction obtains formula

Compound;

C) make the reaction of step a) and step b), obtain formula

Compound;

D) make the reaction product hydrolysis of step c), obtain formula

Compound; With

Wherein: Hal represents halogen for example fluorine, chlorine, bromine, iodine; N is 1, R ₁Definition described with claim 1.

8, the method for preparing the described formula II compound of claim 4, this method may further comprise the steps:

A) in the presence of alkali, make formula With formula

The compound reaction obtains formula Compound;

B) in the presence of alkali, make formula

Compound and methoxychlor methane reaction obtain formula

Compound;

C) in the presence of alkali, make the reaction of step a) and step b), obtain formula

Compound;

D) in The suitable solvent, in the presence of acid, make the reaction product hydrolysis of step c), obtain formula

Compound; With

E) make the reaction product of step d) carry out cyclization and obtain formula II compound; And the optional step of formula II compound being carried out chiral separation.

9, a kind of pharmaceutical composition, it comprises each described compound of claim 1-6 and optional one or more pharmaceutically acceptable carriers or the vehicle that treats and/or prevents significant quantity.

10, each described compound of claim 1-6 preparation be used for the treatment of and/or the medicine of prevention of brain ischemic disease in purposes.

11, each described compound of claim 1-6 is used for the treatment of and/or prevents purposes in the medicine of nerve degenerative diseases in preparation, for example is used for the treatment of and/or prevents purposes in the medicine of Parkinson's disease (Parkinsonism) or alzheimer's disease in preparation.

12, each described compound of claim 1-6 is used for the treatment of and/or prevents purposes in the medicine of inflammatory diseases in preparation, for example preparation be used for the treatment of and/or the medicine of prevention of ulcerative colitis or rheumatoid arthritis in purposes.

13, the purposes of each described compound of claim 1-6 in the preparation medicine, described medicine is used for the next item down or multinomial:

I) agonist of erss hypotype (ER β);

Ii) hypermnesia agent;

Iii) neuroprotective; With

Iv) analgesic agent.