CN108218800A - 1,2,3- triazole aminopeptidase N inhibitor and its preparation method and application - Google Patents
1,2,3- triazole aminopeptidase N inhibitor and its preparation method and application Download PDFInfo
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Abstract
The invention discloses a kind of 1,2,3 triazole aminopeptidase N inhibitors and its preparation method and application.Structure of the compound of the present invention shown in logical formula (I) or (II).Preparation method the present invention also provides 1,2,3 triazole compound and the application in the drug of prevention or treatment with the extremely relevant disease of aminopeptidase activity is prepared.
Description
Technical field
The present invention relates to a kind of 1,2,3- triazole aminopeptidase N inhibitors and its preparation method and application, belong to organic
Compound synthesis and medical applications technical field.
Background technology
Aminopeptidase N (APN, CD13) is zinc ion dependence metalloproteinases, participates in the degradation of substrate N-terminal amino acid, with
The form of homodimer is present in cell membrane, and research has shown that, Aminopeptidase N tumorigenesis, invasion transfer, apoptosis and
It plays an important role in Tumor Angiongesis.(1) Aminopeptidase N is in tumor cell surface high level expression.The enzyme is degradable thin
Extracellular matrix, so as to promote tumor cell invasion and transfer.Metastases are that tumour is difficult to cure major reason.Cell
Epimatrix plays very important effect in the stability of cell connection and intercellular signal conduction is maintained.(2) Aminopeptidase N energy
Enough stimulation vascular endothelial cell release tumor capilaries form correlation factor, promote tumour cell angiogenesis.New vessels energy
It is enough supported for tumor tissue with nutrients, and transports metabolic waste.APN is in neovascular endothelium cell and the high table of sub- endothelial cell
It reaches, its hetero-organization around endothelial cell invasion can be facilitated.Angiogenesis is the first step of growth and metastasis of tumours.(3) Aminopeptidase N
(CD13) be tumor stem cell surface marker, the existence with liver-cancer stem cell has significant relationship (J Clin
Invest2010,120(9),3326-3339).Tumor stem cell is the important original for leading to chemotherapy of tumors drug resistance, recurrence and transfer
Cause.(4) APN participates in the inflammatory reaction that T lymphocytes rely on.Antigen presenting cell surface can be expressed in, immunocompetence of degrading
Substance (such as interleukin-8)；Recognition capability of the T cell surface to its antigen is also reduced, while weakens macrophage and NK is thin
Born of the same parents decline immunity of organisms the identification of tumour cell and killing ability.
Ubenimex is the drug of only one listing up to now, has the class dipeptide structure of beta-amino acids, as exempting from
Epidemic disease reinforcing agent is used for the treatment of leukaemia.It is from the netted streptomycete of olive (Streptomyces olivorecticuli)
It is isolated in culture solution, it is fully synthetic to cost dearly, thus limited source.Now it has been reported that APN inhibitor, it is most of
It is more sensitive to the degradation of internal enzyme for peptide, due to the presence of internal a variety of aminopeptidases, carboxypeptidase or dipeptide peptidase, choosing
Selecting property is poor, it is difficult to develop into the aminopeptidase N inhibitor with clinical value.In addition, peptide inhibitor is multiple due to containing
Chiral centre, synthesis difficulty is big, and greatly mostly from natural products, the limited of source is unfavorable for deep scientific research and future
Large-scale production.Therefore, the aminopeptidase N inhibitor that exploitation is effectively easily-synthesized seems and is highly desirable.
Invention content
In view of the deficiencies of the prior art, the present invention provides a kind of 1,2,3- triazole aminopeptidase N inhibitors and its systems
Preparation Method, the present invention also provides the pharmaceutical applications of the compound.
The technical scheme is that：
First, 1,2,3- triazole aminopeptidase N inhibitors
With the compound of structure shown in below formula (I) or (II) and its optical isomer, diastereoisomer and
Racemic mixture, pharmaceutically acceptable salt, solvate or prodrug：
Wherein,
X is aliphatic group, aryl radical；
R1It is aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocycle, substituted heterocyclic radical；
* it is S or R configurations or its raceme.
According to currently preferred,
Wherein：X is
R1It is phenyl, substituted-phenyl, pyridyl group, naphthalene；
* it is S configurations.
According to the present invention it is further preferred that above compound is one of following：
2nd, the preparation method of 1,2,3- triazole aminopeptidase N inhibitors
The preparation method of 1,2, the 3- triazole aminopeptidase N inhibitors of the present invention is one of following：
(1) preparation method with the compound of logical formula (I) or (II) structure is as follows：
1) leucine methyl ester hydrochloride 1 reacts generation isocyanates with triphosgene, then with propargylamine under alkaline condition
Intermediate 2 is generated, generates intermediate 3 with azanol nak response afterwards；Different aldehyde 4 are by sodium borohydride reduction, Mesylation, Sodium azide
Nucleophilic displacement of fluorine generates intermediate 7；The arylamine 9 of difference substitution generates intermediate 10 by diazotising, Sodium azide nucleophilic displacement of fluorine
(10a-10v)；2- nitrobenzaldehydes 12 are by sodium borohydride reduction, vulcanization sodium reduction generation intermediate 14；Then pass through diazonium
Change, Sodium azide nucleophilic displacement of fluorine, generate intermediate 15；Intermediate 15 and different acylated with acid chloride generation intermediate 10 (10aa-10ii)；
Intermediate 7 or 10 occurs click with intermediate 3 respectively and reacts, and generates target compound 8 or 11；Reaction equation 1 is as follows：
Reagent and condition in above-mentioned reaction equation 1：(a) triphosgene, sodium bicarbonate, dichloromethane, water, 0 DEG C, 1.5 hours；
(b) propargylamine, triethylamine, dichloromethane, 25 DEG C, 12 hours；(c) azanol potassium, methanol, 25 DEG C, 0.5 hour；(d) hydroboration
Sodium, methanol, 0 DEG C, 1 hour；(e) methylsufonyl chloride, dichloromethane, triethylamine, 0 DEG C, 12 hours；(f) Sodium azide, N ' N- diformazans
Base formamide, 25 DEG C, 12 hours；(g) intermediate 3, cupric sulfate pentahydrate, sodium ascorbate, volume ratio 4：1 dimethyl sulfoxide (DMSO)/
Water, 25 DEG C, 2 hours；(h)i：Sodium nitrite, 3.7% hydrochloric acid of mass fraction, 0 DEG C, 0.5 hour, ii：Sodium azide, 0 DEG C, 0.5 is small
When；(i) vulcanized sodium, water, 100 DEG C, 3 hours；(j) various acyl chlorides, triethylamine, anhydrous tetrahydro furan, 50 DEG C, 3 hours.
(2) preparation method with the compound of logical formula (I) structure is as follows：
Aldehydes or ketones 16 are by sodium borohydride reduction, Mesylation, Sodium azide nucleophilic displacement of fluorine generation intermediate 19；Intermediate 19
Click occurs with intermediate 3 and reacts generation target compound 20；Reaction equation 2 is as follows：
Reagent and condition in above-mentioned reaction equation 2：(a) sodium borohydride, methanol, 0 DEG C, 1 hour；(b) methylsufonyl chloride, two
Chloromethanes, triethylamine, 0 DEG C, 12 hours；(c) Sodium azide, N ' dinethylformamides, 25 DEG C, 12 hours；(d) intermediate 3, five
Brochanite, sodium ascorbate, volume ratio 4：1 dimethyl sulfoxide (DMSO)/water, 25 DEG C, 2 hours.
According to currently preferred, the preparation method with the compound of general formula I is as follows：
The preparation of compound 8a-8w, 8aa-8ii and 20a-20e, by taking compound 8a as an example.
(S) -2 (3- ((1- benzyls -1H-1,2,3- triazole-4-yls) methyl) urea groups)-N- hydroxy-4-methyl pentanamides
The preparation method of (8a), step are as follows：
1) preparation of methyl (Propargyl -1- bases carbamoyl)-L-Leu (2)
1 1.81g of L-Leu methyl ester hydrochloride is dissolved in dichloromethane 40mL and the mixing of saturated sodium bicarbonate 40mL is molten
In agent, triphosgene 0.98g being added in by several times, react 1.5 hours at 0 DEG C, point take organic phase, anhydrous magnesium sulfate is dried 15 minutes, is filtered
It crosses, at 0 DEG C, in the anhydrous methylene chloride solution 100mL of filtrate added drop-wise to propargylamine 0.55g and triethylamine 1.21g, drop finishes, and 25
DEG C reaction 12 hours；Concentration, residue are dissolved in ethyl acetate 100mL, and 3.7% hydrochloric acid 100mL of mass fraction is washed three times, unsaturated carbonate
Hydrogen sodium 100mL is washed three times, and saturated sodium-chloride 100mL is washed three times, and anhydrous magnesium sulfate is dried overnight, and is filtered, and concentration obtains crude product；
Crude product recrystallization from ethyl acetate/petroleum ether obtains white solid 1.87g, yield：83%；
2) preparation of (S)-N- hydroxy-4-methyls -2- (3- (Propargyl -1- bases) urea groups) propionamide (3)
The preparation of azanol potassium methanol solution：Potassium hydroxide 28.00g and hydroxylamine hydrochloride 23.35g be dissolved in respectively 70mL and
In the methanol of steaming again of 120mL, solution A and solution B are obtained；Solution A is added drop-wise in solution B, is stirred 40 minutes at 0 DEG C；Filtration, obtains
To the methanol solution of the azanol potassium of brand-new；2 2.26g of compound is dissolved in above-mentioned solution 20mL, and 25 DEG C are reacted 0.5 hour, dense
Contracting, residue are dissolved in water, 3.7% hydrochloric acid tune pH to 6 of mass fraction；Ethyl acetate 100mL is extracted three times, merges organic phase, organic
It is mutually washed three times with saturated sodium bicarbonate 100mL, saturated sodium-chloride 100mL is washed three times, and anhydrous magnesium sulfate drying filters, concentration；It is residual
Slag adds in dichloromethane 50mL, and -18 DEG C stand overnight, and solid is precipitated, and filters, and vacuum drying obtains white solid 1.20g, receives
Rate：53%；
3) preparation of benzyl alcohol (5a)
Benzaldehyde 4a 1.06g are dissolved in methanol 10mL, add in sodium borohydride 0.57g at 0 DEG C by several times, are reacted 1 hour at 0 DEG C,
Concentration, residue are dissolved in ethyl acetate 100mL；Organic phase is washed three times with water 100mL, and saturated sodium-chloride 100mL is washed three times, anhydrous sulphur
Sour magnesium drying, filters, and concentration obtains colorless oil 2.01g, and crude product is direct plungeed into and reacted in next step without being further purified；
The preparation method of compound 5b-5w, 5aa-5ii and 17a-17e are similar with compound 5a；
4) benzyl methanesulfonates (6a)
Compound 5a 1.08g and triethylamine 1.52g are dissolved in anhydrous methylene chloride 100mL, and methylsulfonyl is added dropwise at 0 DEG C
The anhydrous methylene chloride 20mL solution of chlorine 1.36g is reacted 12 hours at 0 DEG C, is concentrated, and residue is dissolved in ethyl acetate 100mL, quality
3.7% hydrochloric acid 100mL of score is washed three times, and saturated sodium bicarbonate 100mL is washed three times, and saturated sodium-chloride 100mL is washed three times, anhydrous sulphur
Sour magnesium is dried overnight, and concentration obtains yellow oil 1.77g；Crude product is direct plungeed into and is reacted in next step without being further purified；
The preparation method of compound 6b-6w, 6aa-6ii and 18a-18e are similar with compound 6a；
5) azido methyl benzene (7a)
Compound 6a 1.86g are dissolved in N ' dinethylformamide 10mL, add in Sodium azide 0.72g, 25 DEG C of reactions 12 by several times
Hour, reaction solution is poured into ice water, and ethyl acetate 100mL is extracted three times, and organic phase is washed three times with water 100mL, saturated sodium-chloride
100mL is washed three times, and anhydrous magnesium sulfate is dried overnight, and concentration obtains colorless oil 1.18g, crude product is without being further purified, directly
Input is reacted in next step；
The preparation method of compound 7b-7w, 7aa-7ii and 19a-19e are similar with compound 7a；
6) ((S) -2 (3- ((1- benzyls -1H-1,2,3- triazole-4-yls) methyl) urea groups)-N- hydroxy-4-methyl pentanamides
(8a)
Compound 3 0.45g and compound 7a 0.29g are dissolved in dimethyl sulfoxide (DMSO) 16mL, add in sodium ascorbate
The water 2mL solution of 59.4mg and the water 2mL solution of cupric sulfate pentahydrate 25mg, 25 DEG C are reacted 2 hours, and reaction solution is poured into water, second
Acetoacetic ester 100mL is extracted three times, merges organic phase, and saturated sodium-chloride 100mL is washed three times, and anhydrous magnesium sulfate is dried overnight, concentration
Obtain colorless oil；Residue is dissolved in dichloromethane 20mL, and 25 DEG C are stirred 2 hours, and the precipitation filtration of generation obtains white solid
0.35g, yield：48%；
The preparation method of compound 8b-8w, 8aa-8ii and 20a-20e are similar with compound 8a.
According to currently preferred, the preparation method with the compound of general formula II is as follows：
(1) preparation method of compound 11a-11v, by taking compound 11a as an example.
(S)-N- hydroxy-4-methyls -2- (3- ((1- phenyl -1H-1,2,3- triazole-4-yls) methyl) urea groups) pentanamide
The preparation method of (11a), step are as follows：
1) triazobenzene (10a)
Aniline 9a 0.93g are dissolved in 3.7% hydrochloric acid of mass fraction, and sodium nitrite 0.74g is added at 0 DEG C, and 0 DEG C of reaction 0.5 is small
When, Sodium azide 0.72g is then added in, 0 DEG C the reaction was continued 0.5 hour, and ethyl acetate 50mL is extracted three times, organic phase anhydrous slufuric acid
Magnesium is dried, and is filtered, and concentration obtains yellow oil, and crude product is direct plungeed into and reacted in next step without being further purified；
The preparation method of compound 10b-10v is similar with compound 10a；
2) (S)-N- hydroxy-4-methyls -2- (3- ((1- phenyl -1H-1,2,3- triazole-4-yls) methyl) urea groups) pentanamide
(11a)
Compound 3 0.45g and compound 10a 0.26g are dissolved in dimethyl sulfoxide (DMSO) 16mL, add in sodium ascorbate 59.4mg
Water 2mL solution and cupric sulfate pentahydrate 25mg water 2mL solution, 25 DEG C react 2 hours, reaction solution is poured into water, ethyl acetate
100mL is extracted three times, merges organic phase, and saturated sodium-chloride 100mL is washed three times, and anhydrous magnesium sulfate is dried overnight, and is concentrated to give nothing
Color oil, residue are dissolved in dichloromethane 20mL, and 25 DEG C are stirred 2 hours, and the precipitation filtration of generation obtains white solid 0.33g, receives
Rate：47%.
The preparation method of compound 11b-11v is similar with compound 11a.
(2) preparation method of compound 11aa-11ii, by taking compound 11aa as an example.
(S) -2- (4- ((3- (1- azanols base) -4- methyl-1s-oxo-pentane -2- bases) urea groups) methyl) -1H-1,2,3- tri-
Azoles -1- bases) benzylacetic acid ester (11aa) preparation method, step is as follows：
1) (2- nitrobenzophenones) methanol (13)
12 1.51g of 2- nitrobenzaldehydes is dissolved in methanol 10mL, adds in sodium borohydride 0.57g at 0 DEG C by several times, is reacted at 0 DEG C
1 hour, concentration, residue was dissolved in ethyl acetate 100mL, and organic phase is washed three times with water 50mL, and saturated sodium-chloride 50mL is washed three times, nothing
Water magnesium sulfate is dried overnight, and filtration is concentrated to give yellow solid 1.45g, crude product direct plunges into next without being further purified
Step reaction；
2) (2- aminophenyls) methanol (14)
13 1.53g of compound is added to the water 100mL solution of vulcanized sodium 21.60g, and 100 DEG C are reacted 3 hours, water phase second
Acetoacetic ester 50mL is extracted three times, and organic phase is washed three times with water 50mL, and saturated sodium-chloride 50mL is washed three times, and anhydrous magnesium sulfate is dried
Night filters, and concentration obtains white solid 1.05g, and crude product is direct plungeed into and reacted in next step without being further purified；
3) (2- azidos phenyl) methanol (15)
14 1.23g of compound is dissolved in 3.7% hydrochloric acid of mass fraction, and sodium nitrite 0.74g, 0 DEG C of reaction 0.5 are added at 0 DEG C
Hour, Sodium azide 0.72g is then added in, 0 DEG C the reaction was continued 0.5 hour, and ethyl acetate 50mL is extracted three times, the anhydrous sulphur of organic phase
Sour magnesium drying, filters, and concentration obtains yellow oil, and crude product is direct plungeed into and reacted in next step without being further purified；
4) 2- azidos benzylacetic acid ester (10aa)
Compound 15 1.49g and triethylamine 1.51g are dissolved in anhydrous tetrahydro furan 50mL, and 0 DEG C is added dropwise chloroacetic chloride 0.94g's
Anhydrous tetrahydro furan 10mL solution, 50 DEG C are reacted 3 hours, and concentration, residue is dissolved in ethyl acetate 50mL, organic phase mass fraction
3.7% hydrochloric acid 50mL is washed three times, and saturated sodium bicarbonate is washed three times, and saturated sodium-chloride is washed three times, and anhydrous magnesium sulfate is dried overnight, filter
It crosses, concentrates, obtain yellow oil 1.07g, crude product is direct plungeed into and reacted in next step without being further purified；
The preparation method of compound 10bb-10ii is similar with compound 10aa；
5) (S) -2- (4- ((3- (1- azanols base) -4- methyl-1s-oxo-pentane -2- bases) urea groups) methyl) -1H-1,2,3-
Triazol-1-yl) benzylacetic acid ester (11aa)
Compound 3 0.45g and compound 10aa 0.42g are dissolved in dimethyl sulfoxide (DMSO) 16mL, add in sodium ascorbate
The water 2mL solution of 59.4mg and the water 2mL solution of cupric sulfate pentahydrate 25mg, 25 DEG C are reacted 2 hours, and reaction solution is poured into water, second
Acetoacetic ester 100mL is extracted three times, merges organic phase, and saturated sodium-chloride 100mL is washed three times, and anhydrous magnesium sulfate is dried overnight, concentration
Colorless oil is obtained, residue is dissolved in dichloromethane 20mL, and 25 DEG C are stirred 2 hours, and the precipitation filtration of generation, vacuum drying obtains white
Color solid 0.44g, yield：53%.
The preparation method of compound 11bb-11ii is similar with compound 11aa.
The target compound structural formula of reaction equation 1 is as follows：
The target compound structural formula of reaction equation 2 is as follows：
The concrete operation step of the compound will be described in detail in embodiment.
Those skilled in the art can change above-mentioned steps to improve yield, they can be according to the basic of this field
Knowledge determines synthetic route, such as selects reactant, solvent and temperature, can be by using various GPF (General Protection False bases to avoid secondary anti-
The generation answered is so as to improve efficiency.These conventional guard methods can be found in such as T.Greene, Protecting Groups
in Organic Synthesis。
3rd, the purposes of 1,2,3- triazole aminopeptidase N inhibitors
The present invention also provides 1,2,3- triazole aminopeptidase N inhibitors to prepare prevention or treatment and aminopeptidase activity
Application in the drug of abnormal relevant disease；Described includes with the extremely relevant disease of aminopeptidase activity：Various blood tumors
Or solid tumor, inflammation, multiple sclerosis, various tissue ulcers or tissue ulcer's venereal disease disease, periodontosis, kabner's disease
Disease, malaria.
In addition, the invention also includes a kind of pharmaceutical composition being suitable for administered orally or parenterally, general formula I or II institutes are included
The compound stated and one or more pharmaceutically acceptable carriers or excipient.
Description of the drawings
Fig. 1 is the representative picture that Human umbilical vein endothelial cells tubular structure is inhibited to be formed.
Fig. 2 is the inhibiting rate column diagram that tubular structure is inhibited to be formed；Statistical analysis,*P<0.05,8v (100 μM) groups pair
Ubenimex (100 μM) group.
Fig. 3 is the representative picture for inhibiting rat aorta ring Angiogenesis.
Fig. 4 is the inhibiting rate column diagram for inhibiting rat aorta ring Angiogenesis；Statistical analysis,*P<0.05,8v
(100 μM) group is to ubenimex (100 μM) group.
Fig. 5 is the representative picture of the lobe of the lung surface nodules of mouse H22 liver cancer Lung metastases.
Fig. 6 is the lobe of the lung surface nodules number column diagram of mouse H22 liver cancer Lung metastases；Statistical analysis,*P<0.05,8v
(60mg/kg) medication group is to blank group, and ubenimex (60mg/kg) medication group is to blank group.
Specific embodiment
With reference to embodiment, the present invention is described further, but not limited to this.
Embodiment 1. (1) compound 8a-8w, the preparation of 8aa-8ii and 20a-20e, by taking compound 8a as an example.
(S) -2 (3- ((1- benzyls -1H-1,2,3- triazole-4-yls) methyl) urea groups)-N- hydroxy-4-methyl pentanamides
The preparation method of (8a), step are as follows：
1) preparation of methyl (Propargyl -1- bases carbamoyl)-L-Leu (2)
L-Leu methyl ester hydrochloride 1 (1.81g, 10mmol) is dissolved in dichloromethane (40mL) and saturated sodium bicarbonate
The in the mixed solvent of (40mL).Triphosgene (0.98g, 3.3mmol) is added in by several times.It is reacted 1.5 hours at 0 DEG C.Divide and take organic phase,
Anhydrous magnesium sulfate is dried 15 minutes.Filtration, at 0 DEG C, filtrate added drop-wise enter propargylamine (0.55g, 10mmol) and triethylamine (1.21g,
In anhydrous methylene chloride (100mL) 12mmol).Drop finishes, and 25 DEG C are reacted 12 hours.Concentration, residue are dissolved in ethyl acetate
(100mL).3.7% hydrochloric acid of mass fraction (3 × 100mL) is washed, and saturated sodium bicarbonate (3 × 100mL) is washed, saturated sodium-chloride (3 ×
It 100mL) washes, anhydrous magnesium sulfate is dried overnight.Filtration, concentration, obtains crude product.Crude product recrystallization from ethyl acetate/petroleum ether, obtains
To white solid 1.87g.Yield：83%, mp：102.9-104.2℃；1H NMR (300MHz, DMSO-d6)：δ 6.37 (d, J=
8.4Hz, 1H), 6.25 (t, J=5.7Hz, 1H), 4.17 (q, J=8.4Hz, 1H), 3.78 (dd, J=5.7Hz, J=2.4Hz,
2H), 3.61 (s, 3H), 3.05 (t, J=2.4Hz, 1H), 1.68-1.54 (m, 1H), 1.47-1.42 (m, 2H), 0.89-0.84
(m, 6H)；ESI-MS m/z 227.4[M+H]+。
2) preparation of (S)-N- hydroxy-4-methyls -2- (3- (Propargyl -1- bases) urea groups) propionamide (3)
The preparation of azanol potassium methanol solution：Potassium hydroxide (28.00g, 509mmol) and hydroxylamine hydrochloride (23.35g,
It 343mmol) is dissolved in respectively in the methanol of steaming again of 70mL and 120mL, obtains solution A and solution B.Solution A is added drop-wise in solution B, and 0
It is stirred 40 minutes at DEG C.Filtration, obtains the methanol solution of the azanol potassium of brand-new.Compound 2 (2.26g, 10mmol) is dissolved in
State solution (20mL).25 DEG C are reacted 0.5 hour.Concentration.Residue is dissolved in water, 3.7% hydrochloric acid tune pH to 6 of mass fraction.Acetic acid second
Ester (3 × 100mL) extracts, and merges organic phase.Organic phase is washed with saturated sodium bicarbonate (3 × 100mL), saturated sodium-chloride (3 ×
100mL) wash.Anhydrous magnesium sulfate is dried.Filtration, concentration.Residue adds in dichloromethane (50mL), and -18 DEG C stand overnight.It is precipitated solid
Body filters, and vacuum drying obtains white solid 1.20g.Yield：53%, mp：124.5-126.2℃；1H NMR(600MHz,
DMSO-d6)：δ 10.68 (s, 1H), 8.80 (s, 1H), 6.24 (t, J=5.7Hz, 1H), 6.16 (d, J=8.4Hz, 1H), 4.05
(q, J=8.4Hz, 1H), 3.78 (dd, J=5.7Hz, J=2.4Hz, 2H), 3.05 (t, J=2.4Hz, 1H), 1.54-1.50
(m, 1H), 1.34-1.32 (m, 2H), 0.89-0.84 (m, 6H)；ESI-MS m/z 226.3[M-H]-。
3) preparation of benzyl alcohol (5a)
Benzaldehyde 4a (1.06g, 10mmol) is dissolved in methanol (10mL), added in by several times at 0 DEG C sodium borohydride (0.57g,
15mmol).It is reacted 1 hour at 0 DEG C.Concentration, residue are dissolved in ethyl acetate (100mL).Organic phase is washed with water (3 × 100mL), is satisfied
It is washed with sodium chloride (3 × 100mL), anhydrous magnesium sulfate drying.Filtration, concentration, obtains colorless oil (2.01g), crude product without into
One step purifies, and direct plunges into and reacts in next step.
The preparation method of compound 5b-5w, 5aa-5ii and 17a-17e are similar with compound 5a.
4) benzyl methyl sulphonic acid ester (6a)
Compound 5a (1.08g, 10mmol) and triethylamine (1.52g, 15mmol) are dissolved in anhydrous methylene chloride
(100mL) is added dropwise to anhydrous methylene chloride (20mL) solution of mesyl chloride (1.36g, 12mmol) at 0 DEG C.12 are reacted at 0 DEG C
Hour.Concentration, residue are dissolved in ethyl acetate (100mL), and 3.7% hydrochloric acid of mass fraction (3 × 100mL) is washed, saturated sodium bicarbonate
(3 × 100mL) is washed, and saturated sodium-chloride (3 × 100mL) is washed, and anhydrous magnesium sulfate is dried overnight.Concentration, obtains yellow oil
(1.77g).Crude product is direct plungeed into and is reacted in next step without being further purified.
The preparation method of compound 6b-6w, 6aa-6ii and 18a-18e are similar with compound 6a.
5) azido methyl benzene (7a)
Compound 6a (1.86g, 10mmol) is dissolved in N ' dinethylformamides (10mL), adds in Sodium azide by several times
(0.72g, 11mmol).25 DEG C are reacted 12 hours.Reaction solution is poured into ice water, ethyl acetate (3 × 100mL) extraction.Organic phase
It is washed with water (3 × 100mL), saturated sodium-chloride (3 × 100mL) is washed, and anhydrous magnesium sulfate is dried overnight.Concentration, obtains colorless oil
(1.18g).Crude product is direct plungeed into and is reacted in next step without being further purified.
The preparation method of compound 7b-7w, 7aa-7ii and 19a-19e are similar with compound 7a.
6) (S) -2 (3- ((1- phenyl -1H-1,2,3- triazole-4-yls) methyl) urea groups)-N- hydroxy-4-methyl pentanamides
(8a)
Compound 3 (0.45g, 2mmol) and compound 7a (0.29g, 2.2mmol) are dissolved in dimethyl sulfoxide (DMSO) (16mL),
Add in water (2mL) solution of sodium ascorbate (59.4mg, 0.3mmol) and the water of cupric sulfate pentahydrate (25mg, 0.1mmol)
(2mL) solution.25 DEG C are reacted 2 hours.Reaction solution is poured into water, ethyl acetate (3 × 100mL) extraction.Merge organic phase, saturation
Sodium chloride (3 × 100mL) is washed, and anhydrous magnesium sulfate is dried overnight.It is concentrated to give colorless oil.Residue is dissolved in dichloromethane (20mL),
25 DEG C are stirred 2 hours.The precipitation filtration of generation, obtains white solid 0.35g.Yield：48%, mp：184.8-186.2℃.1H
NMR(600MHz,DMSO-d6)：δ 10.67 (s, 1H), 8.79 (s, 1H), 7.90 (s, 1H), 7.37 (t, J=6.6Hz, 2H),
7.32 (t, J=6.6Hz, 1H), 7.30 (d, J=6.6Hz, 2H), 6.36 (t, J=6Hz, 1H), 6.11 (d, J=8.4Hz,
1H), 5.56 (s, 2H), 4.21-4.20 (m, 2H), 4.06 (q, J=8.4Hz, 1H), 1.51-1.49 (m, 1H), 1.33-1.30
(m, 2H), 0.86-0.82 (m, 6H)；13C-NMR(100MHz,DMSO-d6)：δ 169.9,157.6,146.6,136.5,129.1,
128.5,128.3,122.9,53.1,49.5,42.9,35.4,24.6,23.2,22.6；HRMS(AP-ESI)m/z calcd
for C17H25N6O3[M+H]+361.1988 found 361.1992.
The preparation method of compound 8b-8w, 8aa-8ii and 20a-20e are similar with compound 8a.
The preparation method of 2. compound 11a-11v of embodiment, by taking compound 11a as an example.
(S)-N- hydroxy-4-methyls -2- (3- ((1- phenyl -1H-1,2,3- triazole-4-yls) methyl) urea groups) pentanamide
The preparation method of (11a), step are as follows：
1) triazobenzene (10a)
Aniline 9a (0.93g, 10mmol) is dissolved in 3.7% hydrochloric acid of mass fraction, added at 0 DEG C sodium nitrite (0.74g,
10.7mmol).0 DEG C is reacted 0.5 hour, then adds in Sodium azide (0.72g, 11mmol).0 DEG C the reaction was continued 0.5 hour.Acetic acid
Ethyl ester (3 × 50mL) extracts, the drying of organic phase anhydrous magnesium sulfate.Filtration, concentration, obtains yellow oil, crude product is without further
Purifying is direct plungeed into and is reacted in next step.
The preparation method of compound 10b-10v is similar with compound 10a.
2) (S)-N- hydroxy-4-methyls -2- (3- ((1- phenyl -1H-1,2,3- triazole-4-yls) methyl) urea groups) pentanamide
(11a)
Compound 3 (0.45g, 2mmol) and compound 10a (0.26g, 2.2mmol) are dissolved in dimethyl sulfoxide (DMSO) (16mL), add
Enter water (2mL) solution of sodium ascorbate (59.4mg, 0.3mmol) and the water (2mL) of cupric sulfate pentahydrate (25mg, 0.1mmol)
Solution.25 DEG C are reacted 2 hours.Reaction solution is poured into water, ethyl acetate (3 × 100mL) extraction.Merge organic phase, saturation chlorination
Sodium (3 × 100mL) is washed three times, and anhydrous magnesium sulfate is dried overnight.It is concentrated to give colorless oil.Residue is dissolved in dichloromethane (20mL),
25 DEG C are stirred 2 hours.The precipitation filtration of generation, obtains white solid 0.33g, yield：47%.1H NMR(300MHz,DMSO-
d6)：δ 10.69 (s, 1H), 8.81 (s, 1H), 8.58 (s, 1H), 7.90-7.87 (m, 2H), 7.62-7.57 (m, 2H), 7.51-
7.46 (m, 1H), 6.48 (t, J=5.7Hz, 1H), 6.18 (d, J=8.7Hz, 1H), 4.34-4.31 (m, 2H), 4.08 (q, J=
8.7Hz, 1H), 1.66-1.43 (m, 1H), 1.37-1.32 (m, 2H), 0.88-0.84 (m, 6H)；13C-NMR (75MHz, DMSO-
d6)：δ 169.4,157.2,146.9,136.6,129.8,128.5,120.6,119.9,49.1,42.3,34.8,24.1,
22.7 22.1；HRMS(AP-ESI)m/z calcd for C16H23N6O3[M+H]+347.1832 found 347.1826.
The preparation method of compound 11b-11v is similar with compound 11a.
The preparation method of 3. compound 11aa-11ii of embodiment, by taking compound 11aa as an example.
(S) -2- (4- ((3- (1- (hydroxylamine) -4- methyl-1s-oxo-pentane -2- bases) urea groups) methyl) -1H-1,2,3-
Triazol-1-yl) benzylacetic acid ester (11aa) preparation method, step is as follows：
1) (2- nitrobenzophenones) methanol (13)
2- nitrobenzaldehydes 12 (1.51g, 10mmol) are dissolved in methanol (10mL), and sodium borohydride is added in by several times at 0 DEG C
(0.57g, 15mmol).It is reacted 1 hour at 0 DEG C.Concentration, residue are dissolved in ethyl acetate (100mL).Organic phase with water (3 ×
It 50mL) washes, saturated sodium-chloride (3 × 50mL) is washed, and anhydrous magnesium sulfate is dried overnight.Filtration, is concentrated to give yellow solid
(1.45g).Crude product is direct plungeed into and is reacted in next step without being further purified.
2) (2- aminophenyls) methanol (14)
Compound 13 (1.53g, 10mmol) is added to water (100mL) solution of vulcanized sodium (21.60g, 90mmol).100
DEG C reaction 3 hours.Water phase is extracted with ethyl acetate (3 × 50mL).Organic phase is washed with water (3 × 50mL), saturated sodium-chloride (3 ×
It 50mL) washes, anhydrous magnesium sulfate is dried overnight.Filtration, is concentrated to give white solid (1.05g).Crude product without being further purified,
It direct plunges into and reacts in next step.
3) (2- azidos phenyl) methanol (15)
Compound 14 (1.23g, 10mmol) is dissolved in 3.7% hydrochloric acid of mass fraction, added at 0 DEG C sodium nitrite (0.74g,
10.7mmol).0 DEG C is reacted 0.5 hour, then adds in Sodium azide (0.72g, 11mmol).0 DEG C the reaction was continued 0.5 hour.Acetic acid
Ethyl ester (3 × 50mL) extracts, the drying of organic phase anhydrous magnesium sulfate.Filtration, concentration, obtains yellow oil, crude product is without further
Purifying is direct plungeed into and is reacted in next step.
4) 2- azidos benzylacetic acid ester (10aa)
Compound 15 (1.49g, 10mmol) and triethylamine (1.51g, 15mmol) are dissolved in anhydrous tetrahydro furan (50mL), and 0
DEG C be added dropwise chloroacetic chloride (0.94g, 12mmol) anhydrous tetrahydro furan (10mL) solution.50 DEG C are reacted 3 hours.Concentration, residue are molten
In ethyl acetate (50mL).Organic phase is washed with 3.7% hydrochloric acid of mass fraction (3 × 50mL), saturated sodium bicarbonate (3 × 50mL)
It washes, saturated sodium-chloride (3 × 50mL) is washed, and anhydrous magnesium sulfate is dried overnight.Filtration, concentration, obtains yellow oil (1.07g).
Crude product is direct plungeed into and is reacted in next step without being further purified.
The preparation method of compound 10bb-10ii is similar with compound 10aa.
5) (S) -2- (4- ((3- (1- azanols base) -4- methyl-1s-oxo-pentane -2- bases) urea groups) methyl) -1H-1,2,3-
Triazol-1-yl) benzylacetic acid ester (11aa)
Compound 3 (0.45g, 2mmol) and compound 10aa (0.42g, 2.2mmol) are dissolved in dimethyl sulfoxide (DMSO) (16mL),
Add in water (2mL) solution of sodium ascorbate (59.4mg, 0.3mmol) and the water of cupric sulfate pentahydrate (25mg, 0.1mmol)
(2mL) solution.25 DEG C are reacted 2 hours.Reaction solution is poured into water, ethyl acetate (3 × 100mL) extraction.Merge organic phase, saturation
Sodium chloride (3 × 100mL) is washed, and anhydrous magnesium sulfate is dried overnight.It is concentrated to give colorless oil.Residue is dissolved in dichloromethane (20mL),
25 DEG C are stirred 2 hours.The precipitation filtration of generation, vacuum drying obtain white solid 0.44g, yield：53%.1H NMR
(300MHz, DMSO-d6)：δ 10.68 (s, 1H), 8.80 (s, 1H), 8.26 (s, 1H), 7.67-7.49 (m, 4H), 6.48 (t, J
=5.7Hz, 1H), 6.19 (d, J=9Hz, 1H), 4.99 (s, 2H), 4.40-4.28 (m, 2H), 4.08 (q, J=9Hz, 1H),
1.96 (s, 3H), 1.60-1.32 (m, 1H), 1.24-1.17 (m, 2H), 0.88-0.84 (m, 6H)；13C-NMR (75MHz, DMSO-
d6)：δ 169.8,169.3,157.1,146.0,135.5,131.1,129.9,129.8,129.3,125.8,124.1,61.6,
49.1,42.3,34.7,24.1,22.7,22.1,20.4；HRMS(AP-ESI)m/z calcd for C19H27N6O5[M+H]+
419.2043 found 419.2039.
The preparation method of compound 11bb-11ii is similar with compound 11aa.
Embodiment 4. inhibits the activity experiment of Aminopeptidase N in vitro
1) experiment material：
Pig source Aminopeptidase N is purchased from Biocol companies, and substrate L- leucyls-p-NA is purchased from Sigma companies.
The preparation of buffer solution, 12.89g Na2HPO4 .12H2O and 2.18g NaH2PO4 .2H2O is dissolved in 1000mL volumetric flasks
In, add and newly boil the distilled water constant volume let cool, obtain the 50mM phosphate buffer solutions that pH is 7.2, be placed at room temperature for spare.
Substrate is dissolved in the solution that 16mmol/mL is made into dimethyl sulfoxide (DMSO), spare.
Aminopeptidase N is dissolved in buffer solution, is made into the solution of 0.15IU/mL, and 4 DEG C of placements of refrigerator are spare.
2) experimental method：
The various addition reagent proportionings of the suppression enzyme experiment in vitro of table 1.
In 96 orifice plates, enzyme, substrate, inhibitor and phosphate buffer are added according to the amount in upper table.In 37 DEG C of incubations
0.5 hour.Absorbance is measured at 405nm, inhibiting rate is calculated according to equation below：
According to the concentration and inhibiting rate of compound, using 8.0 software matched curves of origin, IC is obtained50Value.
3) experimental result：
The Aminopeptidase N of part of compounds and positive drug ubenimex inhibits in logical formula (I) of the present invention and (II) structure
Activity is as shown in table 2 below：
The Aminopeptidase N inhibitory activity of 2. part of compounds of table
[a]Experimental result is expressed as mean+SD, from experiment independent three times.
External suppression enzyme experimental result is shown：Majority of compounds is demonstrated by aminopeptidase in micromole or sub-micromolar level
The inhibitory activity of N.The activity of all compounds is superior to positive drug ubenimex (Bestatin).Wherein, compound 8b, 8e,
8h, 8k, 8l, 8s, 8t, 8w, 8aa, 8bb, 8cc, 11a, 11b, 11e, 11h, 11k-11q, 11s, 11u, 11v, 11aa-11ee,
The activity of 11hh and 20d is better than ubenimex an order of magnitude；The activity of compound 8v, 11ff and 11ii are better than ubenimex two
A order of magnitude.
5. extracorporeal suppression tumor cell proliferation experiment of embodiment
1) experiment material：
Cell strain：
K562 cell strain in exponential phase, human liver cancer PLC/PRF/5 cell strains, human prostata cancer
PC-3 cell strains, people's clear cell carcinoma of ovary ES-2 cell strains, human breast carcinoma A549 cell strains, human breast carcinoma MDA-MB-231 are thin
Born of the same parents' strain, human colon carcinoma HCT116 cell strains.
Reagent：MTT is dissolved in the solution that 1 × PBS is made into 5mg/mL, dimethyl sulfoxide (DMSO).
2) experimental method：
The cell (100 μ L) in growth period of taking the logarithm is laid on 96 orifice plate bottoms (8,000 cells/well of suspension cell, attached cell
4,000 cells/wells), (suspension cell is directly added into inhibitor) adds in the inhibitor of various concentration after the adherent stretching, extension of cell
(100 μ L), 37 DEG C, 5%CO2It is incubated 48 hours.Then MTT (20 μ L) solution is added in, continues to be incubated 4 hours.Centrifugation.Add in two
Methyl sulfoxide (200 μ L), mixing 15 minutes.Absorbance is measured at 570nm.Cell proliferation inhibition rate is calculated according to equation below：
According to the concentration of inhibitor and corresponding inhibiting rate, using 8.0 software matched curves of origin, IC is obtained50Value.
3) experimental result：
Part of compounds and the inhibition kinds of tumors of positive drug ubenimex in logical formula (I) of the present invention and (II) structure
The activity of cell strain proliferation is as shown in table 3 below：
The anti-tumour cell proliferative activity of 3. part of compounds of table
[a]Experimental result is expressed as mean+SD, from experiment independent three times.
Cell-proliferation activity is shown：Compound 8v and 8w is substantially better than the inhibiting effect of seven kinds of cell strains positive drug crow benzene
Department of U.S., has the function of anti-tumour cell proliferative.
6. external anti-angiogenesis of embodiment is tested
(1) Human umbilical vein endothelial cells tubular structure is inhibited to form experiment in vitro
1) experimental model：External Human umbilical vein endothelial cells tubular structure is formed；
2) experiment material：The Human umbilical vein endothelial cells of primary separation, matrigel (Matrigel, purchased from BD
Bioscience companies), M199 cell culture mediums；
3) experimental method：
50 μ L matrigels are added in 96 orifice plates, and 37 DEG C are placed 0.5 hour, and after gelling is solid, it is quiet to add in 50 μ L navels containing people
The M199 culture mediums of arteries and veins endothelial cell (20,000 cells/wells) and various concentration inhibitor.37 DEG C, 5%CO2It is incubated 4 hours.
It micro- Microscopic observation and takes pictures, randomly selects five visuals field and tubular structure branch point is counted, average value is calculated, with black benzene
Department of U.S. group is compared.This experiment is in triplicate.
4) experimental result：
Experimental result is shown in attached drawings 1 and attached drawing 2.The inhibition that compound 8v forms Human umbilical vein endothelial cells tubular structure is in
Existing dose dependent.Compound 8v inhibits the effect that tubular structure is formed with positive drug ubenimex at 100 μM at 10 μM
Quite；Compound 8v almost inhibits the formation of tubular structure at 100 μM.Compound 8v shows external anti-angiogenic life
Into effect, there is the value further developed.
(2) inhibit the experiment of rat aorta ring Angiogenesis in vitro
1) experimental model：External rat aorta ring Angiogenesis；
2) experiment material：The rat of 8-10 week old, matrigel (Matrigel, purchased from BD Bioscience companies), M199
Cell culture medium；
3) experimental method：
Divide and take rat aorta, be cut into the aortic annulus of 1mm or so, it is spare.Matrigel (100 μ L) is added to 96 orifice plates,
37 DEG C are placed 0.5 hour, after gelling is solid, is added in point aortic annulus taken, are then covered matrigel (100 μ L).37 DEG C of placements
0.5 hour, after gelling is solid, add in the inhibitor of various concentration, 37 DEG C, 5%CO2It is incubated 9 days.Micro- Microscopic observation is taken pictures,
Picture is handled with 6.0 softwares of Image-pro Plus, calculates inhibiting rate.Cell culture medium is replaced once every three days.This
Experiment is in triplicate.
4) experimental result：
Experimental result is shown in attached drawings 3 and attached drawing 4.Compound 8v has the inhibiting effect of rat aorta ring Angiogenesis
Dose dependent.Compound 8v inhibits the effect of Angiogenesis with positive drug ubenimex in 100 μM of phases at 10 μM
When；Compound 8v almost inhibits the generation of capilary at 100 μM.Compound 8v shows external anti-angiogenesis
Effect has the value further developed.
Inhibit neoplasm lung metastasis experiment in 7. body of embodiment
1) experimental model：Mouse H22 liver cancer cells Lung metastases；
2) experiment material：The kunming mice of 6 week old, mouse H22 liver cancer cells；
3) experimental method：
Mouse tail vein injection is suspended in the H22 cells (7.5 × 10 of 1 × PBS (0.1mL)6/0.1mL).Injection is completed
Afterwards, mouse is randomly divided into blank group and medication group.Inhibitor or ubenimex, dosage 60mg/kg/d is injected intraperitoneally in medication group.
1 × PBS is injected intraperitoneally in blank group.After medication 12 days, neck is drawn to put to death mouse, divide and take the lobe of the lung, Bouin ' s fixers are fixed, and are counted
The Lung neoplasm on lobe of the lung surface calculates inhibiting rate.
4) experimental result：
Experimental result is shown in attached drawings 5 and attached drawing 6.Compound 8v and ubenimex are in intraperitoneal injection, dosage 60mg/kg/d
When, the inhibiting effect to H22 Lung metastases is respectively 71% and 64%.Compound 8v medication groups have no apparent liver and spleen toxicity.Change
It closes object 8v and shows the effect for inhibiting metastases in vivo, there is further Development volue.
Claims (10)
1.1,2,3- triazole aminopeptidase N inhibitors and its optical isomer, diastereoisomer and raceme mixing
Object, pharmaceutically acceptable salt, solvate or prodrug, with the structure shown in below formula (I) or (II)：
Wherein,
X is aliphatic group, aryl radical；
R1It is aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocycle, substituted heterocyclic radical；
* it is S or R configurations or its raceme.
2. as described in claim 11,2,3- triazole aminopeptidase N inhibitors, it is characterised in that：
Wherein：X is
R1It is phenyl, substituted-phenyl, pyridyl group, naphthalene；
* it is S configurations.
3. as claimed in claim 1 or 21,2,3- triazole aminopeptidase N inhibitors, it is characterised in that for having structure
One of compound：
4. the preparation method of compound as claimed in claim 1 or 2, it is characterised in that be one of following：
(1) preparation method with the compound of logical formula (I) or (II) structure is as follows：
1) leucine methyl ester hydrochloride 1 reacts generation isocyanates with triphosgene, is then generated under alkaline condition with propargylamine
Intermediate 2 generates intermediate 3 with azanol nak response afterwards；Different aldehyde 4 are by sodium borohydride reduction, Mesylation, Sodium azide nucleophilic
Substitution generation intermediate 7；The arylamine 9 of difference substitution is by diazotising, Sodium azide nucleophilic displacement of fluorine, 10 (10a- of generation intermediate
10v)；2- nitrobenzaldehydes 12 are by sodium borohydride reduction, vulcanization sodium reduction generation intermediate 14；Then pass through diazotising, fold
Nitrogen sodium nucleophilic displacement of fluorine generates intermediate 15；Intermediate 15 and different acylated with acid chloride generation intermediate 10 (10aa-10ii)；It is intermediate
Body 7 or 10 occurs click with intermediate 3 respectively and reacts, and generates target compound 8 or 11；Reaction equation 1 is as follows：
Reagent and condition in above-mentioned reaction equation 1：(a) triphosgene, sodium bicarbonate, dichloromethane, water, 0 DEG C, 1.5 hours；(b)
Propargylamine, triethylamine, dichloromethane, 25 DEG C, 12 hours；(c) azanol potassium, methanol, 25 DEG C, 0.5 hour；(d) sodium borohydride, first
Alcohol, 0 DEG C, 1 hour；(e) methylsufonyl chloride, dichloromethane, triethylamine, 0 DEG C, 12 hours；(f) Sodium azide, N ' N- dimethyl methyls
Amide, 25 DEG C, 12 hours；(g) intermediate 3, cupric sulfate pentahydrate, sodium ascorbate, volume ratio 4：1 dimethyl sulfoxide (DMSO)/water, 25
DEG C, 2 hours；(h)i：Sodium nitrite, 3.7% hydrochloric acid of mass fraction, 0 DEG C, 0.5 hour, ii：Sodium azide, 0 DEG C, 0.5 hour；
(i) vulcanized sodium, water, 100 DEG C, 3 hours；(j) various acyl chlorides, triethylamine, anhydrous tetrahydro furan, 50 DEG C, 3 hours；
(2) preparation method with the compound of logical formula (I) structure is as follows：
Aldehydes or ketones 16 are by sodium borohydride reduction, Mesylation, Sodium azide nucleophilic displacement of fluorine generation intermediate 19；Intermediate 19 is in
Click reaction generation target compounds 20 occur for mesosome 3；Reaction equation 2 is as follows：
Reagent and condition in above-mentioned reaction equation 2：(a) sodium borohydride, methanol, 0 DEG C, 1 hour；(b) methylsufonyl chloride, dichloromethane
Alkane, triethylamine, 0 DEG C, 12 hours；(c) Sodium azide, N ' dinethylformamides, 25 DEG C, 12 hours；(d) intermediate 3, five water sulphur
Sour copper, sodium ascorbate, volume ratio 4：1 dimethyl sulfoxide (DMSO)/water, 25 DEG C, 2 hours.
5. the preparation method of compound as claimed in claim 3, (S) -2 (3- ((1- benzyls -1H-1,2,3- triazole-4-yls)
Methyl) urea groups)-N- hydroxy-4-methyls pentanamide (8a) preparation process it is as follows：
1) preparation of methyl (Propargyl -1- bases carbamoyl)-L-Leu (2)
1 1.81g of L-Leu methyl ester hydrochloride is dissolved in the mixed solvent of dichloromethane 40mL and saturated sodium bicarbonate 40mL
In, triphosgene 0.98g is added in by several times, react 1.5 hours at 0 DEG C, point takes organic phase, anhydrous magnesium sulfate is dried 15 minutes, is filtered,
At 0 DEG C, in the anhydrous methylene chloride solution 100mL of filtrate added drop-wise to propargylamine 0.55g and triethylamine 1.21g, drop finishes, and 25 DEG C anti-
It answers 12 hours；Concentration, residue are dissolved in ethyl acetate 100mL, and 3.7% hydrochloric acid 100mL of mass fraction is washed three times, saturated sodium bicarbonate
100mL is washed three times, and saturated sodium-chloride 100mL is washed three times, and anhydrous magnesium sulfate is dried overnight, and is filtered, and concentration obtains crude product；Crude product
With recrystallization from ethyl acetate/petroleum ether, white solid 1.87g, yield are obtained：83%；
2) preparation of (S)-N- hydroxy-4-methyls -2- (3- (Propargyl -1- bases) urea groups) propionamide (3)
The preparation of azanol potassium methanol solution：Potassium hydroxide 28.00g and hydroxylamine hydrochloride 23.35g are dissolved in 70mL's and 120mL respectively
It steams in methanol again, obtains solution A and solution B；Solution A is added drop-wise in solution B, is stirred 40 minutes at 0 DEG C；Filtration, obtains brand-new
Azanol potassium methanol solution；2 2.26g of compound is dissolved in above-mentioned solution 20mL, and 25 DEG C are reacted 0.5 hour, concentration, residue
It is dissolved in water, 3.7% hydrochloric acid tune pH to 6 of mass fraction；Ethyl acetate 100mL is extracted three times, merges organic phase, organic phase saturation
Sodium bicarbonate 100mL is washed three times, and saturated sodium-chloride 100mL is washed three times, and anhydrous magnesium sulfate drying filters, concentration；Residue adds in two
Chloromethanes 50mL, -18 DEG C stand overnight, and solid is precipitated, and filter, and vacuum drying obtains white solid 1.20g, yield：53%；
3) preparation of benzyl alcohol (5a)
Benzaldehyde 4a 1.06g are dissolved in methanol 10mL, add in sodium borohydride 0.57g at 0 DEG C by several times, are reacted 1 hour at 0 DEG C, dense
Contracting, residue are dissolved in ethyl acetate 100mL；Organic phase is washed three times with water 100mL, and saturated sodium-chloride 100mL is washed three times, anhydrous slufuric acid
Magnesium is dried, and is filtered, and concentration obtains colorless oil 2.01g, and crude product is direct plungeed into and reacted in next step without being further purified；
The preparation method of compound 5b-5w, 5aa-5ii and 17a-17e are similar with compound 5a；
4) benzyl methanesulfonates (6a)
Compound 5a 1.08g and triethylamine 1.52g are dissolved in anhydrous methylene chloride 100mL, and mesyl chloride is added dropwise at 0 DEG C
The anhydrous methylene chloride 20mL solution of 1.36g is reacted 12 hours at 0 DEG C, is concentrated, and residue is dissolved in ethyl acetate 100mL, quality point
Several 3.7% hydrochloric acid 100mL are washed three times, and saturated sodium bicarbonate 100mL is washed three times, and saturated sodium-chloride 100mL is washed three times, anhydrous slufuric acid
Magnesium is dried overnight, and concentration obtains yellow oil 1.77g；Crude product is direct plungeed into and is reacted in next step without being further purified；
The preparation method of compound 6b-6w, 6aa-6ii and 18a-18e are similar with compound 6a；
5) azido methyl benzene (7a)
Compound 6a 1.86g are dissolved in N ' dinethylformamide 10mL, add in Sodium azide 0.72g by several times, and 25 DEG C of reactions 12 are small
When, reaction solution is poured into ice water, and ethyl acetate 100mL is extracted three times, and organic phase is washed three times with water 100mL, saturated sodium-chloride
100mL is washed three times, and anhydrous magnesium sulfate is dried overnight, and concentration obtains colorless oil 1.18g, crude product is without being further purified, directly
Input is reacted in next step；
The preparation method of compound 7b-7w, 7aa-7ii and 19a-19e are similar with compound 7a；
6) ((S) -2 (3- ((1- benzyls -1H-1,2,3- triazole-4-yls) methyl) urea groups)-N- hydroxy-4-methyls pentanamide (8a)
Compound 3 0.45g and compound 7a 0.29g are dissolved in dimethyl sulfoxide (DMSO) 16mL, add in sodium ascorbate 59.4mg's
The water 2mL solution of water 2mL solution and cupric sulfate pentahydrate 25mg, 25 DEG C are reacted 2 hours, and reaction solution is poured into water, ethyl acetate
100mL is extracted three times, merges organic phase, and saturated sodium-chloride 100mL is washed three times, and anhydrous magnesium sulfate is dried overnight, and is concentrated to give nothing
Color oil；Residue is dissolved in dichloromethane 20mL, and 25 DEG C are stirred 2 hours, and the precipitation filtration of generation obtains white solid 0.35g, receives
Rate：48%；
The preparation method of compound 8b-8w, 8aa-8ii and 20a-20e are similar with compound 8a.
6. the preparation method of compound as claimed in claim 3, (S)-N- hydroxy-4-methyls -2- (3- ((1- phenyl -1H-1,
2,3- triazole-4-yls) methyl) urea groups) and pentanamide (11a) preparation process it is as follows：
1) triazobenzene (10a)
Aniline 9a 0.93g are dissolved in 3.7% hydrochloric acid of mass fraction, and sodium nitrite 0.74g is added at 0 DEG C, and 0 DEG C is reacted 0.5 hour,
Then Sodium azide 0.72g is added in, 0 DEG C the reaction was continued 0.5 hour, and ethyl acetate 50mL is extracted three times, organic phase anhydrous magnesium sulfate
It is dry, it filters, concentration obtains yellow oil, and crude product is direct plungeed into and reacted in next step without being further purified；
The preparation method of compound 10b-10v is similar with compound 10a；
2) (S)-N- hydroxy-4-methyls -2- (3- ((1- phenyl -1H-1,2,3- triazole-4-yls) methyl) urea groups) pentanamide
(11a)
Compound 3 0.45g and compound 10a 0.26g are dissolved in dimethyl sulfoxide (DMSO) 16mL, add in the water of sodium ascorbate 59.4mg
The water 2mL solution of 2mL solution and cupric sulfate pentahydrate 25mg, 25 DEG C are reacted 2 hours, and reaction solution is poured into water, ethyl acetate 100mL
Extraction three times, merges organic phase, and saturated sodium-chloride 100mL is washed three times, and anhydrous magnesium sulfate is dried overnight, and is concentrated to give colorless oil,
Residue is dissolved in dichloromethane 20mL, and 25 DEG C are stirred 2 hours, and the precipitation filtration of generation obtains white solid 0.33g, yield：
47%；
The preparation method of compound 11b-11v is similar with compound 11a.
7. the preparation method of compound as claimed in claim 3, (S) -2- (4- ((3- (1- azanols base) -4- methyl-1s-oxo
Pentane -2- bases) urea groups) methyl) -1H-1,2,3- triazol-1-yls) benzylacetic acid ester (11aa) preparation process it is as follows：
1) (2- nitrobenzophenones) methanol (13)
12 1.51g of 2- nitrobenzaldehydes is dissolved in methanol 10mL, adds in sodium borohydride 0.57g at 0 DEG C by several times, and reaction 1 is small at 0 DEG C
When, concentration, residue is dissolved in ethyl acetate 100mL, and organic phase is washed three times with water 50mL, and saturated sodium-chloride 50mL is washed three times, anhydrous
Magnesium sulfate is dried overnight, and filtration is concentrated to give yellow solid 1.45g, and crude product direct plunges into next step without being further purified
Reaction；
2) (2- aminophenyls) methanol (14)
13 1.53g of compound is added to the water 100mL solution of vulcanized sodium 21.60g, and 100 DEG C are reacted 3 hours, water phase acetic acid second
Ester 50mL is extracted three times, and organic phase is washed three times with water 50mL, and saturated sodium-chloride 50mL is washed three times, and anhydrous magnesium sulfate is dried overnight,
Filtration, concentration, obtains white solid 1.05g, and crude product is direct plungeed into and reacted in next step without being further purified；
3) (2- azidos phenyl) methanol (15)
14 1.23g of compound is dissolved in 3.7% hydrochloric acid of mass fraction, and sodium nitrite 0.74g is added at 0 DEG C, and 0 DEG C of reaction 0.5 is small
When, Sodium azide 0.72g is then added in, 0 DEG C the reaction was continued 0.5 hour, and ethyl acetate 50mL is extracted three times, organic phase anhydrous slufuric acid
Magnesium is dried, and is filtered, and concentration obtains yellow oil, and crude product is direct plungeed into and reacted in next step without being further purified；
4) 2- azidos benzylacetic acid ester (10aa)
Compound 15 1.49g and triethylamine 1.51g are dissolved in anhydrous tetrahydro furan 50mL, and 0 DEG C is added dropwise the anhydrous of chloroacetic chloride 0.94g
Tetrahydrofuran 10mL solution, 50 DEG C are reacted 3 hours, and concentration, residue is dissolved in ethyl acetate 50mL, organic phase mass fraction
3.7% hydrochloric acid 50mL is washed three times, and saturated sodium bicarbonate is washed three times, and saturated sodium-chloride is washed three times, and anhydrous magnesium sulfate is dried overnight, filter
It crosses, concentrates, obtain yellow oil 1.07g, crude product is direct plungeed into and reacted in next step without being further purified；
The preparation method of compound 10bb-10ii is similar with compound 10aa；
5) (S) -2- (4- ((3- (1- azanols base) -4- methyl-1s-oxo-pentane -2- bases) urea groups) methyl) -1H-1,2,3- tri-
Azoles -1- bases) benzylacetic acid ester (11aa)
Compound 3 0.45g and compound 10aa 0.42g are dissolved in dimethyl sulfoxide (DMSO) 16mL, add in sodium ascorbate 59.4mg's
The water 2mL solution of water 2mL solution and cupric sulfate pentahydrate 25mg, 25 DEG C are reacted 2 hours, and reaction solution is poured into water, ethyl acetate
100mL is extracted three times, merges organic phase, and saturated sodium-chloride 100mL is washed three times, and anhydrous magnesium sulfate is dried overnight, and is concentrated to give nothing
Color oil, residue are dissolved in dichloromethane 20mL, and 25 DEG C are stirred 2 hours, and the precipitation filtration of generation, vacuum drying obtains white solid
0.44g, yield：53%；
The preparation method of compound 11bb-11ii is similar with compound 11aa.
8. any compounds of claim 1-3 are preparing prevention or treatment and the extremely relevant disease of aminopeptidase activity
Application in drug.
9. it is with the extremely relevant disease of aminopeptidase activity as claimed in claim 8：Various blood tumors or solid tumor, inflammation,
Multiple sclerosis, various tissue ulcers or tissue ulcer's venereal disease disease, periodontosis, epidermolysis bullosa, malaria.
10. a kind of be suitable for pharmaceutical composition administered orally or parenterally, comprising the compound described in claim 1,2 or 3 and
One or more pharmaceutically acceptable carriers or excipient.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109608408A (en) * | 2018-12-25 | 2019-04-12 | 河南师范大学 | The preparation method of Paeonol Schiff link 1,2,3- triazole compound with anti-tumor activity |
| CN110038012A (en) * | 2019-05-30 | 2019-07-23 | 济南大学 | Alkaloid compound with 1,2,3- triazole structure segment is preparing the application in angiogenesis promoting medicine |
| CN114181116A (en) * | 2021-11-22 | 2022-03-15 | 山东中医药大学 | D-amino acid derivative aminopeptidase N inhibitor and preparation method and application thereof |
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| CN101817769A (en) * | 2010-04-15 | 2010-09-01 | 山东大学 | Carbamido peptide aminopeptidase N inhibitor and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109608408A (en) * | 2018-12-25 | 2019-04-12 | 河南师范大学 | The preparation method of Paeonol Schiff link 1,2,3- triazole compound with anti-tumor activity |
| CN110038012A (en) * | 2019-05-30 | 2019-07-23 | 济南大学 | Alkaloid compound with 1,2,3- triazole structure segment is preparing the application in angiogenesis promoting medicine |
| CN110038012B (en) * | 2019-05-30 | 2021-06-11 | 济南大学 | Application of alkaloid compound with 1,2, 3-triazole structural segment in preparation of angiogenesis promoting medicine |
| CN114181116A (en) * | 2021-11-22 | 2022-03-15 | 山东中医药大学 | D-amino acid derivative aminopeptidase N inhibitor and preparation method and application thereof |
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