CN105294842A - Cyclic hexadepsipeptide compound and application thereof in preparing benign prostatic hyperplasia resistant drug - Google Patents

Cyclic hexadepsipeptide compound and application thereof in preparing benign prostatic hyperplasia resistant drug Download PDF

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CN105294842A
CN105294842A CN201510725256.3A CN201510725256A CN105294842A CN 105294842 A CN105294842 A CN 105294842A CN 201510725256 A CN201510725256 A CN 201510725256A CN 105294842 A CN105294842 A CN 105294842A
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compound
benign prostatic
prostatic hyperplasia
drug
application
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CN105294842B (en
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鞠建华
宋永相
许芳
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South China Sea Institute of Oceanology of CAS
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South China Sea Institute of Oceanology of CAS
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Abstract

The invention discloses a kind of cyclic hexadepsipeptide compound and an application thereof in preparing a benign prostatic hyperplasia resistant drug. The structural formula of the cyclic hexadepsipeptide compound and a salt thereof for the drug are shown as c(G-W-L-dL-R-N) (formula (I)), wherein in the formula (I), the compound 1: R is equal to Ser/S; compound 2: R is equal to Asp/D; the compound 3: R is equal to Glu/E; the compound 4: R is equal to His/H; or the compound 5: R is equal to Met/M. The cyclic hexadepsipeptide compound has good nonselective anti-junction effect for three hypotypes of alpha1-AR, namely alpha1A-, alpha1B- and alpha1D-AR, for partial hypotypes, the effect is better than a positive reference drug prazosin, and an important significance for developing the benign prostatic hyperplasia resistant drug can be realized..

Description

One class ring hexapeptide compounds and the application in the anti-benign prostatic hyperplasia medicine of preparation thereof
Technical field:
The invention belongs to natural product field, be specifically related to class ring six peptides and preparing α 1-AR antagonist and the application in the anti-benign prostatic hyperplasia medicine of preparation.
Background technology:
Along with the development of society, aging population becomes inevitable social concern, and under population policy situation with Chinese characteristics, old healthy state is related to working order and the work quality of its children, concerns the sound development of society.But, according to statistics, about have 60-70% to suffer from benign prostatic hyperplasia (benignprostatichyperplasia/BPH) in the male sex more than 60 years old, clinical manifestation is the lower urinary tract symptoms such as frequent micturition, urgent urination, enuresis nocturna and misnicturition, has had a strong impact on the quality of life of patient.At present, the common drug of clinical treatment BPH has α 1-AR blocker, 5α-reductase inhibitor and natural product preparation etc.
α 1-AR is alpha adrenergic receptor (α-AR) two type α 1-AR and α 2one in-AR, the dynamic property of its distribution on prostata tissue and functional effect and hyperplasia of prostate is blocked closely related, there will be a known its α at present 1a, α 1b and α 1d tri-hypotypes.α 1a-AR is mainly distributed in prostatic matrix components, accounts for α 170% of-AR; α 1b-AR integrated distribution, in prostatic epithelium, only has a small amount of distribution in matrix; Most α 1d-AR is then distributed in urethral orifice, neck of urinary bladder and sacral region.During benign prostatic hyperplasia, the α in hyperplastic tissue 1-AR concentration can projecting healthy tissues.When sympathetic excitability increases, norepinephrine release increases, the latter and α 1-AR combines, and smooth muscle tension is strengthened, and neck of urinary bladder and intraurethral pressure increase, and voiding resistance increases, thus causes lower urinary tract obstruction symptom.α 1-AR antagonist can alleviate sphincter muscle tensity and hyperplasia of prostate degree thus relief of symptoms, be generally acknowledge at present the choice drug for the treatment of BPH.Conventional clinically have Prazosin, terazosin, exploration Lip river new.
Summary of the invention:
First object of the present invention is to provide a class can as α 1the ring hexapeptide compounds of-AR antagonist and pharmaceutical salts thereof.
Ring hexapeptide compounds of the present invention and pharmaceutical salts thereof, its structure is as shown in formula I:
In formula I, compound 1:R=Ser/S; Or compound 2:R=Asp/D; Or compound 3:R=Glu/E; Or compound 4:R=His/H; Or compound 5:R=Met/M; Or compound R=other conventional amino acid that can protect.
The present inventor has synthesized 5 ring hexapeptide compounds by solid phase synthesis technique, and with crossing MS/MS second order ms, the technology such as 1DNMR, determine that 5 monomers are ring hexapeptide compounds.Concrete structure is as shown in formula I..
By to compound 1-5 and 2 homologous series native annulus hexapeptide compounds 6-7 to α 1the resistive connection activity rating of-AR, finds that it is to α 1three its α of hypotype of-AR 1a, α 1b and α 1d has and selects inhibit activities preferably, has the potentiality developing anti-benign prostatic hyperplasia lead compound.
Therefore second object of the present invention is to provide the application of this ring hexapeptide compounds in the anti-benign prostatic hyperplasia medicine of preparation.
Described anti-benign prostatic hyperplasia medicine is preferably α 1the agonist drug of-AR.
3rd object of the present invention is to provide a kind of anti-benign prostatic hyperplasia medicine, it is characterized in that, includes the compound as shown in formula I as active ingredient of effective amount, or its pharmaceutical salts, and pharmaceutically acceptable carrier.
Described anti-benign prostatic hyperplasia medicine is preferably α 1the agonist drug of-AR.
Ring hexapeptide compounds of the present invention, to test receptor alpha 1three subtype alphas of-AR 1a, α 1b and α 1d has good restraining effect, and its part effect is better than positive control drug thing Prazosin, can as α 1-AR selective antagonist, the lower urinary tract symptoms such as the frequent micturition caused by benign prostatic hyperplasia, urgent urination, enuresis nocturna and misnicturition may be used for the treatment of, therefore the present invention is that the new anti-benign prostatic hyperplasia medicine of exploitation provides compound candidate, has great importance to the new drug development for the treatment of benign prostatic hyperplasia.
Accompanying drawing illustrates:
Fig. 1 is the relative resistive connection potentiality figures of institute's test sample product to three subtype alpha 1A-, α 1B-and α 1D-AR, wherein 1,2,3,4,5,6,7 respectively representation compound 1,2,3,4,5,6,7,1A, 1B, 1D represent α respectively 1a, α 1b and α 1d.
Embodiment:
Following examples further illustrate of the present invention, instead of limitation of the present invention.
Embodiment 1:
The solid phase synthesis of the compound 1-5 as shown in formula I and Structural Identification
In formula I, compound 1:R=Ser/S; Or compound 2:R=Asp/D; Or compound 3:R=Glu/E; Or compound 4:R=His/H; Or compound 5:R=Met/M.
The solid phase synthesis of the compound 1-5 one, as shown in formula I
1. solid phase synthesis
Adopt Solid phase peptide synthssis technology, the amino acid protected with N-α-Fmoc is for raw material, and Fmoc-AA-2CL resin is carrier, the coupling of HBTU method.Fmoc-AA-2CL resin (1mmol) piperidines-DMF (V:V=1:5) is removed Fmoc protecting group, 15 minutes, after DCM and DMF washing, add Fmoc protected amino acid (3mmol) respectively, HBTU (3mmol), DIEA (3mmol) carries out coupling, 30 minutes, after washing, the step of circularly removing Fmoc protecting group-washing-coupling-wash again, until last amino acid couplings terminates, complete the synthesis of whole piece peptide chain [namely respectively according to G-W-L-dL-S-N (glycine-tryptophane-leucine-D-Leu-Ser-Asn), G-W-L-dL-D-N (glycine-tryptophane-leucine-D-Leu-aspartic acid-l-asparagine), G-W-L-dL-E-N (glycine-tryptophane-leucine-D-Leu-L-glutamic acid-l-asparagine), G-W-L-dL-H-N (glycine-tryptophane-leucine-D-Leu-Histidine-l-asparagine), the order of G-W-L-dL-M-N (glycine-tryptophane-leucine-D-Leu-methionine(Met)-l-asparagine) synthesizes five straight line polypeptide].Last to cut peptide reagent TFA: thioanisole: phenol: dithioglycol: polypeptide gets off from cracking vector resin by distilled water (volume ratio 82.5:5:5:2.5:5); and slough all protective materials simultaneously; after 2 hours; the ether 100ml adding 4 DEG C of precoolings makes polypeptide precipitate; centrifugal collecting precipitate; and with washed with diethylether 3 times, vacuum is drained, and obtains straight line polypeptide crude product.
2. purifying
By the crude product polypeptide analysis qualification obtained, crude product preparative reversed-phase liquid chromatography (RP-HPLC) method is purified, with HPLC and MS Analysis and Identification.Detecting HPLC chromatographic column is 250*4.6mm, Kromasil-C18-5um; Mobile phase A: 0.1%TFA/ acetonitrile, Mobile phase B: 0.1%TFA/H2O; Linear eluent gradient: 47%A-72%B; Flow velocity is 1ml/min, and determined wavelength is 220nm; Single injected sampling amount is 10 μ l.After MS qualification is correct, product obtains fine work polypeptide (straight chain six peptide after purifying) through freeze drier.
3. cyclisation
Proceeded in 500ml round-bottomed flask by straight chain six peptide after purifying above, add 200mlDMF as reaction solvent, polypeptide is reacted in rarer solution, and concentration is 10 -3~ 10 -4about M, adds HBTU condensation reagent, is adjusted to pH8 ~ 9 with DIEA, stirring at room temperature, utilizes HPLC to carry out monitoring response situation, next reaction product is cooled, and vacuum concentration is dry.Thick cyclic peptide preparative reversed-phase liquid chromatography (RP-HPLC) method purifying, with HPLC and MS Analysis and Identification, finally obtains target ring hexapeptide compounds (compound 1-5).
Two, the physicochemical data of compound 1-5
Structural analysis test is carried out to compound 1-5, obtains following physico-chemical property data:
Compound 1:c (G-W-L-dL-S-N) white amorphous powder;
1HNMR(400MHz,DMSO)δ10.85(d,J＝1.6Hz,1H),8.52(d,J＝6.4Hz,1H),8.29(d,J＝6.4Hz,1H),8.25(d,J＝8.4Hz,1H),8.03(t,J＝5.6Hz,1H),7.77(d,J＝8.0Hz,1H),7.73(s,1H),7.52(d,J＝8.0Hz,1H),7.37(d,J＝8.4Hz,1H),7.34(d,J＝8.0Hz,1H),7.17(s,1H),7.15(d,J＝2.0Hz,1H),7.07(t,J＝7.2Hz,1H),7.00(d,J＝7.2Hz,1H),4.58–4.55(m,1H),4.44–4.40(m,1H),4.38–4.35(m,1H),4.33–4.28(m,1H),4.10-4.06(m,1H),3.93(dd,J＝16.0,6.4Hz,1H),3.74(dd,J＝11.0,5.2Hz,1H),3.65(dd,J＝11.0,3.6Hz,1H),3.31(dd,J＝16.0,4.8Hz,1H),3.14(dd,J＝14.8,5.0Hz,1H),2.98(dd,J＝14.8,10.0Hz,1H),2.85(dd,J＝16.0,5.0Hz,1H),2.60(dd,J＝16.0,5.6Hz,1H),1.68–1.55(m,2H),1.55–1.39(m,4H),0.91(d,J＝6.0Hz,3H),0.90(d,J＝6.0Hz,3H),0.86(d,J＝6.0Hz,3H),0.84(d,J＝6.0Hz,3H). 13CNMR(100MHz,DMSO)δ173.9,172.6,172.2,171.6,171.5,170.3,169.5,136.6,127.5,124.0,121.4,118.8,118.6,111.8,110.4,61.3,57.1,55.7,52.0,51.5,49.7,43.7,41.7,39.4,37.1,27.8,25.0,24.5,23.4,23.0,22.9,22.0.(+)-HRESIMSm/z[M+H] +calcdforC 32H 47N 8O 8,671.3511；found,671.3504；[M+Na] +cacldforC 32H 46N 8NaO 8,693.3331；found,693.3323.
Compound 2:c (G-W-L-dL-D-N) white amorphous powder;
1HNMR(400MHz,DMSO)δ10.83(d,J＝2.0Hz,1H),8.61(d,J＝7.5Hz,1H),8.39(d,J＝6.0Hz,1H),8.32(d,J＝8.4Hz,1H),8.01(t,J＝5.6Hz,1H),7.74(d,J＝8.0Hz,1H),7.68(s,1H),7.60(d,J＝8.6Hz,1H),7.51(d,J＝8.0Hz,1H),7.32(d,J＝8.0Hz,1H),7.14(s,1H),7.13(d,J＝2.4Hz,1H),7.06(t,J＝7.6Hz,1H),6.98(t,J＝7.6Hz,1H),4.61–4.48(m,2H),4.46–4.35(m,2H),4.13(dd,J＝14.8,6.8Hz,1H),3.88(dd,J＝16.0,6.5Hz,1H),3.31(dd,J＝16.0,4.8Hz,1H),3.13(dd,J＝14.8,4.8Hz,1H),2.98(dd,J＝14.8,9.6Hz,1H),2.84(dd,J＝16.0,4.8Hz,1H),2.76(dd,J＝16.8,4.0Hz,1H),2.65(dd,J＝16.0,5.7Hz,1H),2.56(dd,J＝16.8,8.8Hz,1H),1.65–1.51(m,2H),1.50–1.35(m,4H),0.89(d,J＝6.4Hz,3H),0.88(d,J＝6.8Hz,3H),0.84(d,J＝6.4Hz,3H),0.82(d,J＝6.8Hz,3H). 13CNMR(100MHz,DMSO)δ173.2,172.7,172.2,172.1,171.3,171.3,171.0,169.4,136.6,127.5,124.0,121.4,118.8,118.6,111.8,110.5,55.7,52.5,51.3,50.6,49.7,43.7,42.1,39.4,37.1,36.1,28.09,24.9,24.4,23.1,23.1,22.9,22.3.(+)-HRESIMSm/z[M+H] +calcdforC 33H 47N 8O 9,699.3461；found,699.3452；[M+Na] +cacldforC 33H 46N 8NaO 9,721.3280；found,721.3267.
Compound 3:c (G-W-L-dL-E-N) white amorphous powder;
1HNMR(400MHz,DMSO)δ10.84(d,J＝1.6Hz,1H),8.64(d,J＝6.8Hz,1H),8.34(d,J＝5.2Hz,1H),8.26(d,J＝8.5Hz,1H),7.93(t,J＝4.8Hz,1H),7.73(s,1H),7.71(s,1H),7.62(d,J＝8.4Hz,1H),7.53(d,J＝8.0Hz,1H),7.34(d,J＝8.0Hz,1H),7.14(s,1H),7.14(s,1H),7.07(t,J＝7.2Hz,1H),6.99(t,J＝7.2Hz,1H),4.58–4.53(m,1H),4.43–4.37(m,2H),4.24–4.17(m,1H),4.15–4.09(m,1H),3.91(dd,J＝16.0,4.0Hz,1H),3.31(dd,J＝16.0,4.0Hz,1H),3.14(dd,J＝14.8,4.8Hz,1H),2.99(dd,J＝14.8,10.0Hz,1H),2.87(dd,J＝16.0,5.2Hz,1H),2.66(dd,J＝16.0,5.2Hz,1H),2.30(t,J＝7.6Hz,2H),2.12–2.04(m,1H),1.79–1.72(m,1H),1.60–1.53(m,2H),1.50–1.42(m,4H),0.92(d,J＝6.0Hz,4H),0.91(d,J＝5.6Hz,3H),0.86(d,J＝6.0Hz,6H). 13CNMR(100MHz,DMSO)δ173.8,173.8,173.0,172.2,171.6,171.0,170.9,168.9,136.1,127.0,123.5,120.9,118.3,118.1,111.311,110.0,55.2,52.8,51.9,50.8,49.2,43.2,41.47,40.0,36.6,30.0,27.5,26.0,24.5,24.1,22.6,22.5,22.4,22.0.(+)-HRESIMSm/z[M+H] +calcdforC 34H 49N 8O 9,713.3617；found,713.3604；[M+Na] +cacldforC 34H 48N 8NaO 9,735.3436；found,735.3421.
Compound 4:c (G-W-L-dL-H-N) white amorphous powder;
1HNMR(400MHz,DMSO)δ14.39(s,1H),10.84(d,J＝1.6Hz,1H),9.00(d,J＝1.2Hz,1H),8.75(d,J＝8.4Hz,1H),8.44(d,J＝5.6Hz,1H),8.37(d,J＝8.4Hz,1H),8.09(s,1H),7.83(d,J＝8.4Hz,1H),7.73(d,J＝8.0Hz,1H),7.70(s,1H),7.51(d,J＝8.0Hz,1H),7.32(d,J＝10.0Hz,1H),7.30(s,1H),7.14(s,1H),7.12(d,J＝2.0Hz,1H),7.05(t,J＝7.5Hz,1H),6.98(t,J＝8.0Hz,1H),4.64–4.55(m,2H),4.42–4.34(m,2H),4.00(dd,J＝13.5,7.5Hz,1H),3.85(dd,J＝16.0,6.4Hz,1H),3.31(dd,J＝12.0,4.0Hz,1H),3.27(dd,J＝11.0,4.0Hz,1H),3.14(dd,J＝14.8,4.8Hz,1H),3.00(dd,J＝14.8,10.0Hz,1H),2.93(dd,J＝17.6,6.4Hz,1H),2.86(dd,J＝14.8,4.0Hz,1H),2.68(dd,J＝16.0,5.6Hz,1H),1.56–1.39(m,3H),1.34(t,J＝7.2Hz,2H),1.14–1.03(m,1H),0.88(d,J＝6.0Hz,3H),0.84(d,J＝6.0Hz,3H),0.78(d,J＝6.5Hz,3H),0.75(d,J＝6.5Hz,3H). 13CNMR(100MHz,DMSO)δ172.2,172.0,171.8,170.7,170.5,169.8,168.9,136.08,133.8,129.6,127.1,123.4,120.8,118.3,118.1,117.2,111.3,110.1,55.2,52.2,52.1,50.7,49.2,43.3,41.9,39.9,36.9,27.7,26.0,24.4,23.9,22.6,22.5,22.2,22.1.(+)-HRESIMSm/z[M+H] +calcdforC 35H 49N 10O 7,721.3780；found,721.3781；[M+Na] +cacldforC 35H 48N 10NaO 7,743.3600；found,743.3599.
Compound 5:c (G-W-L-dL-M-N) white amorphous powder;
1HNMR(400MHz,DMSO)δ10.87(s,1H),8.70(d,J＝7.2Hz,1H),8.32(d,J＝8.8Hz,1H),8.29(d,J＝6.5Hz,1H),7.94(t,J＝5.2Hz,1H),7.77(d,J＝8.0Hz,1H),7.71(d,J＝4.0Hz,1H),7.70(d,J＝3.6Hz,1H),7.51(d,J＝8.0Hz,1H),7.32(d,J＝8.0Hz,1H),7.13(d,J＝6.4Hz,1H),7.13(s1H),7.06(t,J＝7.2Hz,1H),6.98(t,J＝7.2Hz,1H),4.59–4.53(m,1H),4.42–4.33(m,2H),4.24–4.16(m,2H),3.88(dd,J＝16.0,6.4Hz,1H),3.35(dd,J＝14.5,4.8Hz,2H),3.13(dd,J＝14.5,4.8Hz,1H),2.99(dd,J＝14.5,10.0Hz,1H),2.85(dd,J＝16.0,5.6Hz,1H),2.65(d,J＝16.0,5.0Hz,1H),2.51(overlapped,1H),2.43(dd,J＝13.2,8.0Hz,1H),2.08-2.03(m,1H),1.89-1.81(m,1H),1.59–1.52(m,2H),1.49–1.39(m,4H),0.90(d,J＝6.0Hz,3H),0.89(d,J＝6.0Hz,3H),0.84(d,J＝6.0Hz,6H). 13CNMR(100MHz,DMSO)δ172.9,172.1,171.6,171.1,170.9,170.9,169.0,136.1,127.0,123.5,120.8,118.3,118.1,111.3,110.0,55.3,52.5,52.0,50.8,49.3,43.2,41.4,38.9,36.7,30.1,29.6,27.5,24.4,24.1,22.5,22.5,22.4,22.1,14.4.(+)-HRESIMSm/z[M+H] +calcdforC 34H 51N 8O 7S,715.3596；found,715.3588；[M+Na] +cacldforC 34H 50N 8NaO 7S,737.3415；found,737.3405.
Analyze known according to above physicochemical data, the structure of compound 1-5 is as formula I.
In formula I, compound 1:R=Ser/S; Or compound 2:R=Asp/D; Or compound 3:R=Glu/E; Or compound 4:R=His/H; Or compound 5:R=Met/M.I.e. compound 1:c (G-W-L-dL-S-N); Compound 2:c (G-W-L-dL-D-N); Compound 3:c (G-W-L-dL-E-N); Compound 4:c (G-W-L-dL-H-N); Compound 5:c (G-W-L-dL-M-N).
Embodiment 2:
By the ring of embodiment 1 hexapeptide compounds-compound 1-5 and homologous series native annulus hexapeptide compounds 6 (c (G-W-L-dL-L-allo-Ile-N)) and compound 7 (c (G-W-L-dL-V-N)) to α 1three subtype alphas of-AR 1A-, α 1B-and α 1D-AR carries out the test of selectivity antagonistic activity.
1. compound is to α 1-AR subtype-selective suppresses situation to be investigated
Strain culturing and plasmid extraction: will (purchased from PromegaProductIDE8471, it contains reporter gene luc2p-CRE to pGL4.29 [luc2P/CRE/Hygro] containing corresponding plasmid; PGL4.74 [hRluc/TK] is purchased from PromegaProductIDE6921, and it contains reporter gene hRluc-TK; EX-A0967-M29 is purchased from GeneCopoeiaProductIDA0967, and it contains α 1the encoding gene of A-AR; EX-Y3321-M29 is purchased from GeneCopoeiaProductIDY3321, and it contains α 1the encoding gene of B-AR; EX-Y2008-M29 is purchased from GeneCopoeiaProductIDY2008, and it contains α 1the encoding gene of D-AR) bacterial strain, join incubated overnight in LB liquid nutrient medium.Plasmid concentration is measured with Micro Core acid albumin quantitative instrument after extracting plasmid.
2. cell cultures, bed board
By HEK293 cell recovery, proceed in 50mL Tissue Culture Flask, add containing 10% foetal calf serum, 1% dual anti-DMEM high glucose medium, cultivate also every 24h for 37 DEG C and change a nutrient solution, when cell confluency degree reaches 90%, with PBS buffer solution for cleaning cell bottle, trysinization, and with the dilution of 10 times of volume medium, centrifugally abandon supernatant.Cell blows open, by 10 with appropriate DMEM substratum 5individual/ml, spreads 96 orifice plates, in 37 DEG C of constant temperature culture 24h.
3. transient transfection
1) mixed liquor A and the B of fresh cell transfecting is configured
A is by plasmid α 1a (EX-A0967-M29) or α 1b (EX-Y3321-M29) or α 1d (EX-Y2008-M29), reporter gene luc2p-CRE (pGL4.29 [luc2P/CRE/Hygro]) and hRluc-TK (pGL4.74 [hRluc/TK]) and the dual anti-high glucose medium DMEM of serum-free formulated, B by liposome and DMEM formulated.Wherein α 1-AR hypotype eukaryon expression plasmid: reporter gene: internal reference=1:1:1.96 orifice plates, the amount that every hole adds DNA is 0.2ug/25ul, requires that the amount adding liposome is 0.5ul/25ul.
2) transfection
After bed board 24h, each hole totally degree of converging reaches more than 90%, reaches and wink turns requirement.Prepare A and B liquid respectively by above-mentioned calculated amount, leave standstill 5-10 minute, then that A and B is miscible, after liquid-transfering gun mixing, leave standstill 20-30min.From thermostat container, take out Tissue Culture Plate again, after cleaning 2 times with PBS, discard waste liquid, in every hole, add DMEM substratum 50uL respectively, after treating A and B effect, therefrom draw in 50uL to every hole, shake up, put into thermostat container and cultivate, carry out transfection.After cultivating, 4h takes out Tissue Culture Plate, is replaced into by enchylema containing serum and dual anti-DMEM high glucose medium, then cultivates 18-22h.
4. the configuration of testing sample and Activity determination
1 ~ 2mg testing sample constant volume is become by testing compound and phyenlephrinium (agonist) storing solution of 2mmol/L with DMSO.Take out Transfected cells culture plate, clean 2 times with PBS.In every hole, add 99 μ L containing 10% foetal calf serum and 1% dual anti-DMEM high glucose medium, get testing compound respectively and phyenlephrinium 0.5 μ L joins every hole containing in the flat board of 99 μ L substratum (making medicine final concentration be 10 μMs of ol/L).First add antagonist and blank reagent, after effect 30min, then add agonist.Mix enchylema after interpolation, cultivate 8 hours for 37 DEG C.In this experiment, clinical anti-benign prostatic hyperplasia Nonselective antagonists Prazosin (Prazosin) is added to and turns (drug level unification is 10 μMs) in clone wink containing three hypotype plasmids, add phyenlephrinium to stimulate, with the signal suppressing degree by phyenlephrinium, investigate the activity intensity of antagonist, add medicine irritation after 8 hours, detect relative antagonistic vigor (relativeinhibitionpotency/RIP), i.e. ratio (the RLU of surveyed Prazosin Photinus pyralis LUC and renilla luciferase activity, relativelightunits) with the RLU difference of testing sample and the ratio of Prazosin RLU, more small sample activity is better for ratio.
Relative antagonistic vigor (RIP)=(Prazosin RLU value-testing sample RLU value)/Prazosin RLU value
Active results of stimulation is as shown in table 1 and Fig. 1.
A table 1 test sample product are to three subtype alphas 1A-, α 1B-and α 1Dthe RIP value (measuring average 3 times) of-AR
From table 1 and Fig. 1, compound 1-7 is to three of α 1-ARs hypotype: α 1A-, α 1B-, and α 1D-AR has good non-selective antagonistic activity, and the activity of institute's test sample product and positive control are all at an order of magnitude, and compound 1 is to the resistive connection of three hypotypes, compound 3,6 pairs of subtype alphas 1B-, and α 1Dthe resistive connection of-AR, compound 7 couples of α 1A-and α 1Dthe resistive connection of-AR, and compound 2,5 pairs of subtype alphas 1Dthe resistive connection of-AR is all better than positive control Prazosin, based on this, for the research and development carrying out going deep into α 1-ARs subtype selective antagonists provide support.
In sum, the invention provides α 1the non-selective inhibitor of-ARs three hypotypes, on this basis, has great importance for developing the anti-benign prostatic hyperplasia medicine for the treatment of further.

Claims (7)

1. a class ring hexapeptide compounds, or its pharmaceutical salts, its structure is as shown in formula I:
c(G-W-L-dL-R-N)
Formula I
Compound 1:R=Ser; Or compound 2:R=Asp; Or compound 3:R=Glu; Or compound 4:R=His; Or compound 5:R=Met.
2. the application of ring hexapeptide compounds according to claim 1 in the anti-benign prostatic hyperplasia medicine of preparation.
3. application according to claim 2, is characterized in that, described anti-benign prostatic hyperplasia medicine is α 1the antagonist of-AR.
4. application according to claim 3, is characterized in that, described α 1the antagonist of-AR is α 1a, α 1b and/or α 1the antagonist of D.
5. an anti-benign prostatic hyperplasia medicine, is characterized in that, includes the ring hexapeptide compounds according to claim 1 as active ingredient of effective amount, or its pharmaceutical salts, and pharmaceutically acceptable carrier.
6. anti-benign prostatic hyperplasia medicine according to claim 5, is characterized in that, described anti-benign prostatic hyperplasia medicine is α 1the antagonist of-AR.
7. anti-benign prostatic hyperplasia medicine according to claim 6, is characterized in that, described α 1the antagonist of-AR is α 1a, α 1b and/or α 1the antagonist of D.
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