CN103848778B - A kind of aminopeptidase N inhibitor and preparation method thereof and application - Google Patents
A kind of aminopeptidase N inhibitor and preparation method thereof and application Download PDFInfo
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Abstract
The invention discloses a kind of aminopeptidase N inhibitor, chemical name is: 2-((S)-3-(3-((S)-2-amino-3-hydrocinnamyl) urea)-2,6-dioxopiperidin-1-base)-N-glycoloyl amine hydrochlorate, there is the compound of structure formula I:
also disclose the preparation method of this compound and the application in antitumor drug preparation.The aminopeptidase N inhibitor that the present invention obtains, spatially matches with the avtive spot of Aminopeptidase N, therefore all demonstrates stronger anti-tumor activity in antitumor activity evaluation in vivo and in vitro.
Description
Technical field
The present invention relates to a kind of preparation method of aminopeptidase N inhibitor and the medicinal use at anti-tumor aspect, belong to technical field of chemistry.
Background technology
Aminopeptidase N (APN, CD13) people's alanyl-amino peptidase (EC3.4.11.2) is typically referred to, it is the dependent hydrolysising protease of a kind of zine ion, belong to metalloprotease M1 family, being present on cytolemma with the form of homodimer, is same material with cell-surface antigens CD13; Aminopeptidase N has expression on the various kinds of cell surface of Various Tissues, especially abundant in the intestinal epithelial cells of small intestine and the epithelial cell of kidney proximal tubule, in liver, placenta and uterus, also have expression.Aminopeptidase N can from the neutral amino acids of peptide, acid amides or fragrant acid amides hydrolysis release N end or basic aminoacids (Ala>Phe>Tyr>Le u>Arg>Thr>Trp), the natural substrate of effect comprises: blood vessel function peptide, neuropeptide hormone, cytokine and immunomodulator etc.Research proves, Aminopeptidase N plays an important role in tumorigenesis, Invasion and Metastasis, tumor-blood-vessel growth, Immunity regulation and virus infection.
1) APN expresses at tumor cell surface height, can degradation of cell epimatrix, and participates in the generation of tumor neovasculature as a novel signal transduction molecule; thus promote tumor cell invasion and transfer (Bauvois; B.etal.MedResRev., 2006,26 (1): 88; Zhang, X.etal.Curr.Med.Chem., 2008,15 (27): 2850.).2) APN participates in the inflammatory reaction that T lymphocyte relies on, and can be expressed in antigen presenting cell surface, degraded immunologic active material (as interleukin-8); Also reduce the recognition capability of T cell to its antigen, slackened identification and the kill capability of scavenger cell and NK cells against tumor cells simultaneously, immunity of organisms is declined.3) APN is by the chemokine Fmlp that HIV auxiliary receptor CCR 5 can be made to desensitize that degrades, thus reduces the innate immune function of cell, and makes CCR5 enhanced sensitivity; promote HIV cell entry host cell (ShenW.etal.Blood; 2000,96 (8), 2887; ShippMA, etal.Blood, 1993,82 (4), 1052.).4) APN as human corona virus HCoV-229E and Transmissible gastroenteritis virus (TGEV) surface acceptor, play an important role in upper respiratory tract infection (as: SARS) and acute enteritis.（Delmas,B.etal.Nature,1992,357,417;Yeager,CL.etal.Nature,1992,420.）。5) APN can degrade Angiotensin, participates in the adjustment (Mitsui, T.etal.Biol.Pharm.Bull., 2004,27 (6): 768.) of body blood pressure.
Bestatin, commodity are called ubenimex, it is a class dipeptide compound with beta-amino acids structure, be at present unique one listing as the APN inhibitor of immunostimulant for leukemia treating, owing to being be separated to obtain from the nutrient solution of the netted streptomycete of olive (Streptomycesolivorecticuli), limited source.Therefore, utilize chemosynthesis means to obtain the small molecules APN inhibitor with anti-tumor activity to have great importance.
Alpha-amido acyl-ring imide peptoid metalloprotease inhibitor and application thereof is reported in patent (CN200910020656.9); wherein compound 13f has stronger inside and outside anti-tumor activity; but the peptide bond containing easy body endoproteinase degraded in compound 13f structure, internal metabolism less stable.The chemical structural formula of compound 13f is as follows:
Summary of the invention
For the deficiencies in the prior art, the object of this invention is to provide a kind of aminopeptidase N inhibitor, this compound suppresses to suppress in lotus H22 hepatoma mice Blood route metastasis experimental test in the activity experiment of Aminopeptidase N, the test of inhibition tumor cell proliferation activity and body in vitro, and its activity is all better than compound 13f.
Another object of the present invention is to provide the preparation method of this aminopeptidase N inhibitor and is preparing the application in antitumor drug.
For achieving the above object, the present invention adopts following technical scheme:
A kind of aminopeptidase N inhibitor, chemical name is: 2-((S)-3-(3-((S)-2-amino-3-hydrocinnamyl) urea)-2,6-dioxopiperidin-1-base)-N-glycoloyl amine hydrochlorate, there is the compound of structure formula I:
The preparation method of described compound, reactions steps and reaction formula as follows:
1) protect L-glutamine (1) for starting raw material with Boc; intermediate (4) is protected to obtain through cyclization, nucleophilic substitution, de-Boc; intermediate (4) prepares key intermediate isocyanic ester (5) under triphosgene effect, and synthetic route is:
Reagent in said synthesis route and reaction conditions are: (a) dicyclohexylcarbodiimide, N-hydroxysuccinimide, tetrahydrofuran (THF), 65 DEG C; (b) salt of wormwood, tetrabutylammonium iodide, potassiumiodide, acetone, benzyl acetate bromide, 56 DEG C; (c) trifluoroacetic acid, methylene dichloride; (d) triphosgene, sodium bicarbonate, methylene dichloride.
2) (S)-2-tertiary butyloxycarbonyl amide group-3-phenyl-propanol (6); intermediate (9) is obtained after Mesylation, azide, hydro-reduction; intermediate (9) and isocyanic ester (5) condensation obtain urea groups intermediate (10); intermediate (10) is again through hydro-reduction debenzylation; with oxammonium hydrochloride condensation; finally slough Boc protecting group, obtain target compound (ZX-1), synthetic route is:
Reagent in said synthesis route and reaction conditions are: (a) methylsulfonyl chloride, triethylamine, tetrahydrofuran (THF), 0 DEG C; (b) sodium azide, DMF, 65 DEG C; (c) magnesium, methyl alcohol; (d) methylene dichloride, 0 DEG C; (e) 10% palladium charcoal, methyl alcohol, hydrogen; (f) isobutyl chlorocarbonate, triethylamine, oxammonium hydrochloride, 0 DEG C; (g) 3mol/L hydrogenchloride/ethyl acetate solution.
Those skilled in the art can change to improve yield to above-mentioned steps; they can determine the route of synthesis according to the ABC of this area; as selective reaction thing, solvent and temperature, can by using various GPF (General Protection False base to avoid the generation of side reaction thus to improve yield.The guard method of these routines can see such as T.Greene, ProtectingGroupsinOrganicSynthesis.
The aminopeptidase N inhibitor that the present invention obtains, spatially matches with the avtive spot of Aminopeptidase N, and therefore all demonstrate stronger anti-tumor activity in antitumor activity evaluation in vivo and in vitro, this compound is expected to develop into a kind of new type antineoplastic medicine.
Accompanying drawing explanation
Target compound anti-tumor in vivo cell Lung metastases experiment lung surface nodules number prepared by Fig. 1 embodiment of the present invention 1;
Target compound anti-tumor in vivo cell Lung metastases inhibiting rate prepared by Fig. 2 embodiment of the present invention 1.
Embodiment
The present invention is further illustrated in conjunction with the embodiments, should be noted that following explanation is only to explain the present invention, not limiting its content.
The synthesis of embodiment 1. the compounds of this invention (I)
1) preparation of (S)-tertiary butyl-2,6-dioxopiperidin-3-base carbonyl acid amides (2)
By Boc-L-glutamine (9.86g, 40.0mmol) with N-hydroxy-succinamide (4.6g, 40.0mmol) be dissolved in 100ml tetrahydrofuran (THF) (THF), under cryosel bath condition, slowly drip the THF solution that 50ml contains dicyclohexylcarbodiimide (8.24g, 40mmol).Within about 1 hour, drip off, remove ice bath, stirring at room temperature is after 3 hours, back flow reaction 10 hours.Reaction solution is cooled to room temperature, removes solvent under reduced pressure.Residue adds 40ml ethyl acetate reconcentration once.After final residue thing adds 50ml ethyl acetate, refrigerator freezing is placed and is spent the night.With 2.0g diatomite filtration, filtrate is respectively with water (20ml × 1) and saturated brine (20ml × 1) washing, and anhydrous magnesium sulfate drying, filters, be concentrated into dry.White solid 6.8g is obtained again, yield: 74.5%, mp:213-214 DEG C (literature value 212-214 DEG C) with ethyl acetate-ethyl ether recrystallization.
2) preparation of (S)-benzyl-2-(3-(tertiary butyloxycarbonyl amide group)-2,6-dioxopiperidin-1-base) acetic ester (3)
By compound 2(4.0g, 17.5mmol), K
2cO
3(2.9g, 21mmol), tetrabutylammonium iodide (0.07g, 0.18mmol) DL is in 50ml anhydrous propanone.Benzyl acetate bromide (6.0g, 26.3mmol) is slowly added under room temperature condition.Reaction solution refluxes 5 hours, is cooled to room temperature.Filter, concentrate to obtain yellow oil.Add 50ml ethyl acetate, organic phase is respectively with 0.5%Na
2cO
3(20ml × 2), 1N citric acid (20ml × 2), saturated brine are washed till neutrality, and organic phase, with anhydrous magnesium sulfate drying, is filtered, concentrated, obtains compound 3, white solid 4.41g, yield: 67.0% after ethyl acetate-ethyl ether recrystallization.mp：78.0-81.0℃；ESI-MSm/z[M+1]
+：377.6。
3) preparation of (S)-benzyl-2-(3-isocyanic ester-2,6-dioxopiperidin-1-base) acetic ester (5)
By compound 3(2.26g, 6.0mmol) be dissolved in 24ml methylene dichloride, until completely dissolved, slowly drip 6.0mlTFA, about 20min adds, and stirring at room temperature 3 hours, concentrates reaction solution, then adds 20ml methylene dichloride evaporate to dryness.Residue is dissolved in the mixed solution of 30mL/30mL methylene dichloride and saturated sodium bicarbonate, under condition of ice bath, once adds triphosgene (0.6g, 2.0mmol) vigorous stirring afterwards, stop after 1 hour stirring, separatory; Use dichloromethane extraction aqueous phase afterwards three times, merge organic phase, anhydrous magnesium sulfate drying, filter, concentrated solvent, to about 20mL, without the need to further separation and purification, can be directly used in next step.
4) preparation of the synthesis (7) of (S)-2-tertiary butyloxycarbonyl amide group-3-phenyl-methylsulfonic acid propyl ester
By (S)-2-tertiary butyloxycarbonyl amide group-3-phenyl-propanol (6) (11.3g, 45.0mmol) be dissolved in 100mL anhydrous tetrahydro furan, ice bath is cooled to 0 DEG C, add anhydrous triethylamine (4.55g, 45.0mmol), dropwise add 20mL under condition of ice bath and be dissolved with methylsulfonyl chloride (5.15g, anhydrous tetrahydrofuran solution 45.0mmol), within about 0.5 hour, dropwise, remove ice bath, room temperature reaction is after 4 hours, reaction solution impouring is filled in the beaker of cold water, a large amount of white solid is had to separate out, solid filtering is dry, obtain target compound compound 12.0g, yield: 81.0%, mp:105-108 DEG C.
5) preparation of the synthesis (8) of (S)-2-tertiary butyl (2-azido-methyl-1-styroyl) carbamate
By compound 7(10.0g, 30.0mmol) be dissolved in 80mLDMF, be warming up to 60 DEG C, add sodiumazide (3.95g in batches, 60.0mmol), isothermal reaction is after 10 hours, by reaction solution impouring frozen water, extraction into ethyl acetate (50mL × 3), merge organic phase, after anhydrous magnesium sulfate drying, filtering and concentrating obtains crude product, column chromatographic isolation and purification (eluent: petrol ether/ethyl acetate=10/1) obtains target compound 5.0g, yield: 60.3%.
6) preparation of the synthesis (9) of (S)-2-tertiary butyloxycarbonyl amide group-3-phenyl-propylamine
By compound 8(4.0g, 15.0mmol) be dissolved in 80mL anhydrous methanol, under condition of ice bath, add the magnesium rod (1.08g of chopping in batches, 45.0mmol), react after 1 hour, solvent under reduced pressure is steamed and removes, residue adds cold water dilution, with ether extraction (50mL × 3), merges organic phase, saturated aqueous common salt (40mL × 3) washs, anhydrous magnesium sulfate drying, filters, by filtrate evaporate to dryness, obtain white solid 1.75g, yield: 46.6%, mp:114-116 DEG C.ESI-MSm/z：251.1[M+H]
+；
1H-NMR(600MHz,DMSO-d
6)δ1.26-1.33(m,9H),2.62(d,J=6.7Hz,1H),2.63-2.76(m,1H),2.95-3.10(m,1H),3.51(s,1H),3.63(s,1H),6.63-6.70(m,1H),7.17-7.20(m,2H),7.26(s,2H)。
7) preparation of benzyl-2-((S)-3-(3-((S)-2-tertiary butyloxycarbonyl amide group-3-hydrocinnamyl) urea groups)-2,6-dioxopiperidin-1-bases)-acetic ester (10)
By compound 9(1.50g, 6.0mmol) be dissolved in 20mL methylene dichloride, drip 20mL compound 5(6.0mmol in condition of ice bath downhill reaction liquid) dichloromethane solution, after dropwising, continue room temperature reaction 1 hour.Evaporated under reduced pressure solvent, residue is dissolved in 40mL ethyl acetate, successively with 1N citric acid (20mL × 1), saturated brine (20mL × 1) washing, anhydrous magnesium sulfate drying, filter, filtrate decompression evaporate to dryness is obtained crude product, and crude product obtains white solid 1.75g after column chromatographic isolation and purification (eluent: petrol ether/ethyl acetate=4/1), yield 52.8%.mp：168-170℃。ESI-MSm/z：575.3[M+Na]
+1。H-NMR(600MHz，DMSO-d
6)δ1.23-1.28(m,9H),1.86-1.92(m,1H),2.05-2.07(m,1H),2.61-2.65(m,1H),2.71-2.74(m,2H),2.88-2.92(m,1H),2.93-2.98(m,1H),3.15-3.18(m,1H),3.61-3.62(m,1H),4.41-4.49(m,2H),4.52-4.56(m,1H),5.13-5.17(m,2H),6.19-6.24(m,1H),6.55(d,J=6.7Hz,1H),6.70(d,J=6.7Hz,1H),7.17-7.21(m,3H),7.26-7.28(m,2H),7.33-7.40(m,5H)。
8) preparation of 2-((S)-3-(3-((S)-2-tertiary butyloxycarbonyl amide group-3-hydrocinnamyl) urea groups)-2,6-dioxopiperidin-1-bases)-acetic acid (11)
By compound 10(1.50g, 2.72mmol) be dissolved in the anhydrous methanol of 30mL, add catalytic amount 10%Pd/C, with H
2as hydrogen donor, room temperature reaction 16 hours, after reacting completely, filters, obtains compound 11, white solid 1.21g, yield: 96.2%, mp:104-106 DEG C after concentrating under reduced pressure.ESI-MSm/z：485.2[M+Na]
+，
1H-NMR(600MHz，DMSO-d
6)δ1.25-1.33(m,9H),1.83-1.91(m,1H),2.04-2.06(m,1H),2.61-2.64(m,1H),2.69-2.72(m,2H),2.85-2.91(m,1H),2.94-2.98(m,1H),3.14-3.18(m,1H),3.59-3.62(m,1H),4.23-4.31(m,2H),4.48-4.52(m,1H),6.19-6.26(m,1H),6.55(d,J=6.7Hz,1H),6.72(d,J=6.7Hz,1H),7.17-7.20(m,3H),7.26-7.28(m,2H)。
9) preparation of the tertiary butyl-(S)-1-(3-((S)-1-(2-azanol-2-oxygen ethyl)-2,6-dioxopiperidin-3-bases) urea groups)-3-hydrocinnamyl-2-carboxylicesters (12)
By compound 11(0.93g, 2.0mmol) be dissolved in 30mL anhydrous tetrahydro furan, control solution temperature not higher than-20 DEG C, slowly add 0.26mL(2.2mmol) N-methylmorpholine, slowly add 0.3mL(2.2mmol after 5min) isobutyl chlorocarbonate.Reaction solution continues stirring after 30 minutes at-20 DEG C, is slowly dropped in mixed anhydride reaction liquid by oxammonium hydrochloride (0.17g, the 2.45mmol) solution (adjusting pH to neutral with N-methylmorpholine in advance) being dissolved in 3mL anhydrous methanol, dropwises.Continue-20 DEG C of insulation reaction 1 hour, then room temperature reaction 4 hours.After reaction terminates, with 2.0g diatomite filtration, concentrated, add 50mL ethyl acetate, successively with saturated sodium bicarbonate (10mL × 3), 1mol/L citric acid (10mL × 3), saturated aqueous common salt (10mL × 2) washs, anhydrous magnesium sulfate drying.Filter, concentrating under reduced pressure, column chromatography for separation (eluent: petrol ether/ethyl acetate=1/1) obtains compound 12, white solid 0.65g, yield: 68.1%; Mp=74-76 DEG C.ESI-MSm/z：500.2[M+Na]
+，
1H-NMR(600MHz,DMSO-d
6)δ1.26-1.33(m,9H),1.88-1.93(m,1H),2.03(s,1H),2.63-2.66(m,1H),2.70-2.74(m,2H),2.81-2.86(m,1H),2.97(s,1H),3.15-3.17(m,1H),3.61(s,1H),4.12-4.20(m,2H),4.47-4.54(m,1H),6.22(s,1H),6.47(s,1H),6.68(s,1H),7.17-7.20(m,H),7.26-7.28(m,2H),8.80(s,1H),10.53(s,1H)。
10) preparation of 2-((S)-3-(3-((S)-2-amino-3-hydrocinnamyl) urea groups)-2,6-dioxopiperidin-1-bases)-N-hydroxy imide hydrochloride (ZX-1)
By compound 12(0.5g, 1.05mmol) be dissolved in 10mL hydrogenchloride/ethyl acetate (3mol/L) solution, room temperature reaction 3 hours, product is separated out with the form of hydrochloride, filters, and anhydrous diethyl ether washs, be drying to obtain target compound, called after ZX-1, white solid powder 0.36g, yield: 83.7%; Mp=88-90 DEG C.ESI-MSm/z：378.1[M+Na]
+，
1H-NMR(600MHz,DMSO-d
6)δ1.91-1.95(m,1H),2.02-2.05(m,1H),2.67-2.73(m,1H),2.82-2.85(m,2H),2.89-2.95(m,1H),3.14-3.18(m,1H),3.22-3.25(m,1H),3.42-3.45(m,1H),4.13-4.19(m,2H),4.46-4.56(m,1H),6.69(s,1H),6.71-6.74(m,1H),7.26-7.28(m,1H),7.30-7.31(m,2H),7.33-7.36(m,2H),8.14(s,3H),8.78(s,1H),10.60(s,1H)。
Embodiment 2. target compound suppresses the activity experiment (Invitro) of Aminopeptidase N
The test description of Aminopeptidase N (APN) inhibit activities in .Biochemistry such as Lejczak, B, 1989,28, in 3549.Substrate L-leucyl-p-NA is degraded by APN, produces the p-NA having absorption at 405nm, and the size of the concentration of p-NA and enzymic activity is proportionate.By detect 405nm place the content of optical density determination p-NA, thus determine the activity of aminopeptidase, indirectly reflect the size of inhibitor to inhibition of enzyme activity degree.
Concrete grammar and operation steps are see CN100560568C " cyclin imide peptidyl metalloprotease inhibitor and application thereof ".Experimental result is in table 1.
The external Inhibiting enzyme activity test-results of table 1 target compound
In a table, numerical value is the mean value of three tests, the numeric representation standard deviation after " ± ".
Above-mentioned test result shows, target compound all demonstrates to the APN of kidney derived APN and the ES-2 cell expressing of pig the activity being better than positive control bestatin and lead compound 13f in suppressing APN enzymic activity to be tested in vitro.
Embodiment 3. target compound inhibition tumor cell proliferation activity test (Invitro)
The In Vitro Anti proliferation assay to tumour cell of compound adopts Thiazolyl blue detection method (mtt assay), the cell suspension of human oophoroma cell line (ES-2) or Breast cancer lines (MDA-MB-231) is inoculated in 96 orifice plates respectively, the substratum containing different concns compound is added in every hole, after hatching, with MTT dyeing, after continuing to hatch, in the absorbancy (OD value) in the every hole of mensuration, 570nm place in microplate reader, calculate inhibitory rate of cell growth, thus the activity of deterministic compound.
1. [material] ES-2 cell strain, MDA-MB-231 cell strain, Methyl thiazoly tetrazolium assay MTT, 10% foetal calf serum, 96 orifice plates.
2. [method]
Cell cultures clear cell carcinoma of ovary ES-2 cell strain and mammary cancer MDA-MB-231 cell strain all adopt cellar culture.Logarithmic phase cell is all used during experiment.
Growth of Cells detects (mtt assay) and ES-2 cell and MDA-MB-231 cell suspension is all adjusted to 1 × 10
5/ ml, is inoculated in 96 orifice plates (50 μ l/ hole), 5 × 10 respectively
3individual cells/well.After bed board 4h, the substratum of 50 μ l containing different concns compound is added in every hole, final compound concentration in hole is respectively: 4000,800,160,32,6.4 μm of ol/L, each concentration establishes three multiple holes, do not add the hole reading of cell as blank, add the hole that cell do not add compound and make compound blank well, Bestatin makes compound positive control.In 37 DEG C, hatch 48h in 5% carbonic acid gas, every hole adds the MTT staining fluid of 10 μ l0.5%, and after continuing to hatch 4h, 2500rpm, centrifugal 30min, then abandon substratum in plate hole, add dimethyl sulfoxide (DMSO), 200 μ l/ holes.Microplate reader measures the absorbancy OD value in every hole in 570nm place, inhibitory rate of cell growth is calculated as follows:
Inhibiting rate (%)=(control wells mean OD value-experimental port mean OD value)/control wells mean OD value × 100%
According to concentration and the corresponding inhibiting rate of compound, utilize origin7.5 software matching beneficial effect curve, calculate the IC of compound
50. experimental result is in table 2.
Table 2. target compound extracorporeal anti-tumor cell proliferation experiment result
In a table, numerical value is the mean value of three tests, the numeric representation standard deviation after " ± ".
Upper table test data shows, demonstrates the activity being better than positive control bestatin and lead compound 13f, have good DEVELOPMENT PROSPECT in the test of target compound anti-tumour cell proliferative in vitro.
Embodiment 4. target compound suppresses lotus H22 hepatoma mice Blood route metastasis test (Invivo)
Concrete grammar and operation steps are see CN1528745A " pyrrolidinyl metalloprotease inhibitor and preparation method thereof ".Experimental result is shown in Fig. 1 and Fig. 2.
Experimental result shows, target compound demonstrates the activity being better than positive control bestatin and lead compound 13f in the test of suppression lotus H22 hepatoma mice Blood route metastasis, has good DEVELOPMENT PROSPECT.
Above-described embodiment is the present invention's preferably embodiment; but embodiments of the present invention are not restricted to the described embodiments; change, the modification done under other any does not deviate from spirit of the present invention and principle, substitute, combine, simplify; all should be the substitute mode of equivalence, be included within protection scope of the present invention.
Claims (2)
1. a preparation method for aminopeptidase N inhibitor, is characterized in that, comprises the following steps:
1) protect L-glutamine (1) for starting raw material with Boc; intermediate (4) is protected to obtain through cyclization, nucleophilic substitution, de-Boc; intermediate (4) prepares inter-mediate isocyanate (5) under triphosgene effect, and synthetic route is:
Reagent in said synthesis route and reaction conditions are: (a) dicyclohexylcarbodiimide, N-hydroxysuccinimide, tetrahydrofuran (THF), 65 DEG C; (b) salt of wormwood, tetrabutylammonium iodide, potassiumiodide, acetone, benzyl acetate bromide, 56 DEG C; (c) trifluoroacetic acid, methylene dichloride; (d) triphosgene, sodium bicarbonate, methylene dichloride;
2) (S)-2-tertiary butyloxycarbonyl amide group-3-phenyl-propanol (6); intermediate (9) is obtained after Mesylation, azide, hydro-reduction; intermediate (9) and isocyanic ester (5) condensation obtain urea groups intermediate (10); intermediate (10) is again through hydro-reduction debenzylation; with oxammonium hydrochloride condensation; finally slough Boc protecting group, obtain target compound, synthetic route is:
Reagent in said synthesis route and reaction conditions are: (a) methylsulfonyl chloride, triethylamine, tetrahydrofuran (THF), 0 DEG C; (b) sodium azide, DMF, 65 DEG C; (c) magnesium, methyl alcohol; (d) methylene dichloride, 0 DEG C; (e) 10% palladium charcoal, methyl alcohol, hydrogen; (f) isobutyl chlorocarbonate, triethylamine, oxammonium hydrochloride, 0 DEG C; (g) 3mol/L hydrogenchloride/ethyl acetate solution.
2. the application of aminopeptidase N inhibitor in the medicine preparing anti-breast cancer that prepare of preparation method according to claim 1.
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