CN101531613A - Aminopeptidase N inhibitor bestatin dino ester, synthesis and application thereof - Google Patents
Aminopeptidase N inhibitor bestatin dino ester, synthesis and application thereof Download PDFInfo
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- CN101531613A CN101531613A CN200910020655A CN200910020655A CN101531613A CN 101531613 A CN101531613 A CN 101531613A CN 200910020655 A CN200910020655 A CN 200910020655A CN 200910020655 A CN200910020655 A CN 200910020655A CN 101531613 A CN101531613 A CN 101531613A
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Abstract
The invention provides an intensive antitumor medicament, namely bestatin deanol ester, a preparation method and application thereof. The bestatin deanol ester not only can effectively inhibit ectopic expression of aminopeptidase N activity, but also greatly improve the water solubility of the parent drug bestatin of the bestatin deanol ester, has slow release effect, is more excellent than the parent drug bestatin either from healing effect or preparation, and has wide application. Particularly, the invention mainly relates to the following three aspects: (1) design and synthesis of the bestatin deanol ester; (2) N'N deanol which is a micromolecule fragment, can be connected with other medicaments containing carboxyl, improves the pharmacokinetic property of the medicaments, and improves water solubility; and (3) a method for synthesizing an ester substance to which the invention relates with wide application and mild condition.
Description
Technical field
The present invention relates to the prodrug of aminopeptidase N inhibitor Bestatin (ubenimex), the design of ubenimex enlightening promise ester, synthetic and possible medicinal use.Belong to technical field of pharmaceuticals.
Background technology
1. Aminopeptidase N (APN)
Aminopeptidase N (APN/CD13) is a kind of two shaped metal exopeptidases that depend on zine ion, is under the jurisdiction of M1 aminopeptidase extended familys.Its physiologic function is the degraded that participates in substrate N terminal amino acid.Under the normal condition, APN is extensive low expression level in tissues such as mammiferous kidney, small intestine and central nervous system, participates in the physiological regulation of body.Studies have shown that Aminopeptidase N is a kind of multifunctional enzyme, substrate is extensive, and it takes place in tumour, immunologic function is regulated, and virus infection and ease pain all play an important role.
(1) APN is at the tumor cell surface high level expression.This enzyme can make the main component degraded of extracellular matrix (ECM), has destroyed the natural cover for defense of body, promotes invasion by tumor cells, growth and transfer, and participates in the tumour neovascularity and generate, thereby provides required nutrient for growth of tumor.(referring to Saiki, I.; Etal.Int.J.Cancer., 1993,54,137; Sato, Y.Biol.Pharm.Bull., 2004,772,776).
(2) APN has also participated in the inflammatory reaction that the T lymphocyte relies on simultaneously in granulocyte and lymphocytic cell surface great expression; Can also be expressed in the antigen presenting cell surface, degraded immunologic active material (as interleukin-8); The T cell that major histocompatibility complex II type (MHC-II) the Adhesion Antigen determinant of processing of participation antigen and cell surface relies on is to antigenic identification, reduced the recognition capability of T cell, weakened scavenger cell and NK cell identification and kill capability simultaneously, immunity of organisms is descended tumour cell.
(3) APN is as the acceptor on human corona virus HCoV-229E and Transmissible gastroenteritis virus (TGEV) surface, in upper respiratory tract infection (as: SARS) and acute enteritis, play an important role, and it plays a role relevant (referring to Delmas with the activity of enzyme, B., et al.Nature, 1992,357,417; Yeager, C.L.; Etal.Nature, 1992,357,420).
(4) APN participates in the process that the HIV virion enters host cell.Studies show that the APN activity is higher than the healthy volunteer far away in patient's body of infected by HIV.When HIV-1 invasion host cell, the APN of high expression level can make the chemokine fMLP of HIV-1 auxiliary receptor CCR 5 desensitization by degraded, thereby reduces the natural immunity function of cell, and makes the CCR5 enhanced sensitivity, promotes that virus enters host cell.(referring to ShenW, Li B, et al.Blood, 2000,96 (8), 2887; Shipp MA, et al.Blood, 1993,82 (4), 1052)
(5) APN participates in the degraded of endogenous analgesic matter endorphin and enkephalin, thereby causes the excessive release of P material, causes pain.
(6) APN degraded Angiotensin, and the adjusting of participation body blood pressure (referring to Mitsui, T.; Et al.Biol.Pharm.Bull., 2004,27,768.)
2. ubenimex
It is glad that ubenimex has another name called hundred scholars, English Bestatin, Ubenimex by name, and chemistry is by name: nitrogen-[(2S, 3R)-3-amido-2-hydroxy-4-phenyl butyryl]-the L-leucine.(referring to H.Suda et al., ibid.100.Syntheses:eidem, ibid.600; R.Nishizawa et al., ibid.1983,36,695) the CAS number of registration is 58970-76-6.
Ubenimex is a kind of aminopeptidase N inhibitor, and this product can be used for raise immunity, is used for the assisting therapy of anticancer chemotherapy, radiotherapy, senile immunodeficiency etc., also can be directly used in tumour such as leukemic treatment.Its chemical structure is as follows:
Ubenimex
Comprise a primary amine groups and a carboxyl in the ubenimex structure, both can form inner salt, therefore are insoluble in various organic solvents, and water-soluble also relatively poor, bioavailability is low, metabolic stability is poor thereby cause, and has limited its use.
Summary of the invention
The present invention is directed to the deficiency of existing anticancer ancillary drug ubenimex aspect pharmacokinetic property, a kind of good water solubility, good drug efficacy, ubenimex enlightening promise ester that medicine stability is good are provided.
The present invention also provides the synthetic method and the application of above-mentioned ubenimex enlightening promise ester.
In order to improve the ubenimex pharmacokinetic property, improve water-solublely, the present invention transforms ubenimex, and a kind of ubenimex enlightening promise ester is provided, and its structural formula is as follows:
Contain a primary amine groups and a tertiary amine groups in this prodrug structure, both each can be in conjunction with the hydrogenchloride of a part, thereby form bimolecular hydrochloride, water-solublely improve greatly, thus applicable to multiple drug formulation.Make molecular structure more stable simultaneously.
Used term and definition implication is as follows:
Naphthene group is replacement or unsubstituted, saturated or undersaturated cyclic group, and it contains carbon atom and one or more heteroatoms.This ring can be monocycle or condensed ring, the ring system of bridged ring or volution.
THF: tetrahydrofuran (THF);
(Boc)
2O: tert-Butyl dicarbonate, structure is:
HOBt:N-hydroxy benzo tetrazole, structure is:
DCC: dicyclohexylcarbodiimide, structure is:
DMAP: to dimethylamino pyridine, structure is:
The synthetic method of ubenimex enlightening promise ester of the present invention, step is as follows:
(1) ubenimex with optical purity is a starting raw material, under the alkaline condition of pH=9-10, makes the reaction of ubenimex and tert-Butyl dicarbonate, and the primary amine groups of ubenimex is carried out the Boc protection, then,
(2) make solvent with anhydrous tetrahydro furan, under the katalysis of 0 ℃ of temperature and DCC, the ubenimex and the N-hydroxy benzo tetrazole of the product B oc of step (1) protection is condensed into active ester, then,
(3) go up step reaction product active ester and at room temperature reach under the katalysis of DMAP again, be reacted into ester with N ' N dimethylethanolamine, generate the prodrug of Boc protection, the mol ratio of described active ester and N ' N dimethylethanolamine is 1:3.
The prodrug of the Boc protection that (4) step (3) is obtained is taken off the Boc blocking group in the saturated ethyl acetate solution of hydrogenchloride, get target compound.
Wherein, it is as follows that step (3) becomes the ester reaction formula:
R1, R2 are various types of substituting groups, as fat group or aromatic group, can be that chain also can be a cyclic substituents.
Preferred consumption is as follows:
In the step (1), ubenimex is 1:1.1 with the molar weight ratio of tert-Butyl dicarbonate,
In the step (2), N-hydroxy benzo tetrazole is 1:1.08 with the molar weight ratio of ubenimex, and dicyclohexylcarbodiimide is 1:1.1 with the molar weight ratio of ubenimex.
In the step (3), to the dimethylamino pyridine catalytic amount 0.2 times of ubenimex molar weight.
Below being preferred, is a concrete route of synthetic its prodrug of starting raw material with the ubenimex of optical purity:
Contain amido, two active groups of hydroxyl in the ubenimex structure, carry out on this basis amido Boc protection, carboxyl activation, esterification, take off amido Boc and protect this four basic steps, N ' N dimethylethanolamine and carboxyl are connected to ester.Both protect carboxyl, strengthened the metabolic stability of medicine.Meanwhile introduce a tertiary amine groups again, thus can with two molecule hydrogenchloride salifies, increased the water-soluble of medicine, improved its pharmacokinetic property.One-tenth ester method of the present invention has the advantage that does not influence the intramolecularly reactive group.
The present invention designs and has synthesized a ubenimex enlightening promise ester with brand new.Experiment in vitro shows that it presses down enzymic activity significantly, and can become one is the novel prodrugs of skeleton with the ubenimex.This method also is applicable to any design that contains the prodrug of free carboxy medicine.Simultaneously, the synthetic used one-tenth ester method of this prodrug also has novelty, is applicable to that most carboxyls react with the ester that becomes of alcohol, and the reaction conditions gentleness, is particularly useful for the one-tenth ester reaction to thermally labile acid.
From reaction result, this ubenimex enlightening promise ester finally is that the form with two molecule hydrochlorides exists, and has extremely strong water-solublely, helps the preparation of solution in the process of active testing and the design of pharmaceutical dosage form.
Next step will carry out external antitumor cell experiment and vitro stability experiment the ubenimex enlightening promise ester that obtains.
In sum, these invention major technique characteristics comprise following three parts:
(1) this natural radioactivity small molecules and ubenimex that contains the tertiary amine structure forms ester class prodrug with N ' N dimethylethanolamine, thereby improves its stability and pharmacokinetics character.
(2) simultaneously, N ' N dimethylethanolamine also can be used for connecting the compound that other comprise free carboxy, improves that it is water-soluble.
(3) used a kind of one-tenth ester method of novelty in this building-up process, the advantage of these one-tenth ester means is can not have influence on other reactive group in the drug molecule, and the reaction conditions gentleness is widely applicable.
The present invention becomes ester with ubenimex with N ' N dimethylethanolamine, when increasing medicine stability, has improved greatly that it is water-soluble, has well improved its waistband kinetic property, has improved bioavailability, and meta-bolites also has no side effect.Draw inferences about other cases from one instance, any have carboxyl, a water-soluble relatively poor medicine, all can form a kind of structure of ester with N ' N dimethylethanolamine, again with various organic or inorganics acid salifies, increases that it is water-soluble.
N ' N dimethylethanolamine, English N by name, N-dimethyl ethanolamine or Deanol are a kind of natural active matters that exists at nature, it can increase the intravital choline amount of people, thus with memory, study, thinking has confidential relation, (referring to Alfredo Gragnan, etc.
Plast.Surg.2007,31,711-718; AlfredoGragnani, etc.Aesth.Plast.Surg.2008,32,406.) it takes as a kind of medicine in American-European countries, is used for the improvement of crease-resistant and brain function, and toxic side effect is little.Simultaneously, how tumour patient can follow various miseries during treating, thereby makes tumour patient have the fidgets, and as the precursor of choline, N ' N dimethylethanolamine can effectively improve this dysphoric undesirable condition.Therefore the present invention selects N ' N dimethylethanolamine to become ester with ubenimex, this method can not make various sensitive groups in the drug molecule as: amido, hydroxyl, sulfydryl etc. are affected, and the reaction conditions gentleness, have general suitability.
Press down the enzymic activity experiment in the external and body of ubenimex enlightening promise ester of the present invention:
1. external inhibition activity experiment to Aminopeptidase N:
Experimental principle: APN suppresses active test description in Lejczak, and .Biochemistry such as B are in 1989,28,3549.Substrate is L-leucyl-p-N-methyl-p-nitroaniline, and it can be degraded by APN, be created on the p-N-methyl-p-nitroaniline that 405nm has absorption, and the size of the concentration of p-N-methyl-p-nitroaniline and enzymic activity is proportionate.Determine the content of p-N-methyl-p-nitroaniline by the optical density that detects 405nm place, can determine the activity of Aminopeptidase N, thereby reflect the size of inhibitor indirectly enzymic activity inhibition degree
Materials and methods:
Aminopeptidase N and substrate L-leucyl-p-N-methyl-p-nitroaniline are all available from Sigma company
The preparation of solution:
Damping fluid, preparation pH7.2, the 50mM phosphate buffered saline buffer, room temperature is placed standby.
Aminopeptidase N is dissolved in the solution that is made into 0.2mg/mL in the damping fluid;
Substrate is dissolved in and is made into 0.5mg/mL solution among the DMSO, and each solution refrigerator is placed standby.
Amino-peptidase activity is analyzed:
Add above-mentioned Aminopeptidase N solution 10 μ L in 96 orifice plates, substrate solution is supplied 200 μ L with damping fluid., under 35 ℃, in shaking table, rocked 30 minutes, utilize microplate reader to measure absorption value in 405nm wavelength place.
Pressing down enzyme detects:
Add above-mentioned Aminopeptidase N solution 10 μ l in 96 orifice plates respectively, substrate 5 μ l, the compound 40 μ l of different gradient concentrations, compound always has 6 concentration gradients, is respectively 130 μ g/ml, 32 μ g/ml, 8 μ g/ml, 2 μ g/ml, 0.5 μ g/ml and 0.125 μ g/ml.Supply 200 μ L with 175ml phosphate buffered saline buffer (pH7.2) at last.100% group does not contain inhibitor.Blank group replaces organized enzyme with the enzyme of inactivation, all supplies 200 μ L with damping fluid.Under 35 ℃, in shaking table, rocked 30 minutes, utilize microplate reader again, measure each hole absorption value in 405nm wavelength place.Calculate inhibiting rate according to following formula at last:
Inhibiting rate=(100% optical density-compound optical density)/(100% optical density-blank optical density)
According to compound concentrations and corresponding inhibition ratio, utilize Origin7.5 software match beneficial effect curve, calculate the IC of prodrug
50Value.
Table 1: the external enzyme experimental result that presses down
| IC 50(μ mol/ml) value | |
| Ubenimex enlightening promise ester | 0.06 |
| Ubenimex | 0.003 |
Annotate: numerical value is the mean value of three experiments in the table.Enzymic activity experiment shows, the prodrug inhibitory enzyme activity is less than parent drug, ubenimex.
2. vitro inhibition tumor cell viability.
Experimental principle and step:
Thiazolyl blue, be called for short MTT, but permeate through cell membranes enters in the cell, amber desaturase in the viable cell plastosome can make exogenous MTT be reduced to be insoluble in the hepatic Formazan crystallization of water and be deposited in the cell, crystallisate can be dissolved by dimethyl sulfoxide (DMSO) (DMSO), measure its absorbance value with enzyme-linked immunosorbent assay instrument at 570nm wavelength place, can reflect cell quantity indirectly.The amount that the MTT crystallisate forms is directly proportional with viable count, and is also proportional with cell viability.
The test cell is human lung carcinoma cell (A549), clear cell carcinoma of ovary cell (ES-2), and human leukemia cell (HL-60) adds 10% foetal calf serum with the RPMI1640 substratum and cultivates.
Step:
(1) shop 96 orifice plates, every hole 50ul, 5000 cells (A549, ES-2 are 5000, and HL-60 is 50000);
(2) behind the bed board 4h, add the substratum that 50ul contains the different concns compound in the hole, make that the compound final concentration is respectively in the hole: prodrug is 200,100,50,25,12.5ug/ml; Bestatin is 800,600,400,200,100ug/ml, and each concentration is established three multiple holes, does not make Blank when not adding the hole reading of cell, and the compound blank is made in the solvent hole;
(3) behind the adding compound 48h, Xiang Kongzhong adds 0.5% MTT, every hole 10ul;
(4) behind the adding MTT 4h, 2500rpm, 30min is centrifugal, throws plate and abandons substratum in the hole, adds DMSO, the 100ul/ hole;
(5) microplate reader is surveyed 570nm place absorbance, 630nm wavelength for referencial use
Inhibiting rate=(the blank group-test group of the compound)/blank group of compound * 100%
Experimental result:
Table 2: prodrug is to A549 cell inhibiting result
| Inhibiting rate 100% | 12.5ug/ml | 25ug/ml | 50ug/ml | 100ug/ml | 200ug/ml |
| LYP | 4.079277 | 7.056127 | 14.88414 | 32.19397 | 50.16533 |
Table 3: prodrug is to ES-2 cell inhibiting result
| Inhibiting rate 100% | 12.5ug/ml | 25ug/ml | 50ug/ml | 100ug/ml | 200ug/ml |
| LYP | 6.397573 | 26.9612 | 0.990101 | 50.03816 | 61.91936 |
Table 4: prodrug is to HL60 cell inhibiting result
| Inhibiting rate 100% | 50ug/ml | 100ug/ml | 150ug/ml | 200ug/ml |
| LYP | 47.65403 | 40.76252 | 32.55137 | 15.54254 |
Table 5: prodrug, ubenimex are to the median effective dose of three kinds of tumour cells
As can be seen from the above results, ubenimex enlightening promise ester has better effect aspect these three kinds of tumour cells of inhibition.Median effective dose will be much smaller than parent drug---ubenimex.This result's reliability will more be higher than the enzymic activity experiment.
3. experiment in the mouse body
This experiment has selected for use mouse to experimentize, and mouse is inoculated H22 (human liver cancer cell) solid tumor, carries out activity experiment respectively with ubenimex enlightening promise ester and ubenimex, and last is the activity that standard is judged compound with the tumour inhibiting rate.
Ubenimex enlightening promise ester has selected three dosages to be respectively 100mg/kg, 50mg/kg and 25mg/kg, ubenimex concentration is 100mg/kg, shown in experimental result Fig. 1, from then in the histogram as can be seen, prodrug is at 00mg/kg, and the tumour inhibiting rate under three dosage of 50mg/kg and 25mg/kg is respectively 35%, 16% and 13%.
Ubenimex enlightening promise ester and ubenimex specific activity are seen Fig. 2, and under 100mg/kg dosage, the inhibiting rate of ubenimex enlightening promise ester and ubenimex is respectively 35% and 3%, and the activity of ubenimex enlightening promise ester will be apparently higher than ubenimex.
4. mouse interior medicine dynamics test
Get Kunming kind healthy mice, be divided into 2 groups at random, the 1st group is the ubenimex group, and the 2nd group is the prodrug group, 3 of the parallel settings of each time point.Tail vein injection ubenimex and prodrug, ubenimex dosage 100mg/kg, prodrug dosage are then by waiting administration of mole ubenimex dosage.Mouse fasting 12h before the administration freely drinks water, and gets blood respectively at 5min, 15min, 30min, 45min, 1,2,4,6,8,12,24h after (0h), the administration before the administration in the eyeground vein clump, adds in the centrifuge tube of heparin sodium rinse.Accurate absorption blood plasma 200 μ l place the 1.5ml centrifuge tube, the accurate acetonitrile solution 50 μ l that contain 25% perchloric acid that add, and the vortex vibration, centrifugal (15000rpm) 20min gets supernatant sample introduction 25 μ l HPLC mensuration Plasma Concentration respectively.
Average medicine-time curve of ubenimex and prodrug is seen Fig. 3,4,5,6.
Calculate its main statistical moment parameter, the results are shown in Table 6, table 7.By area under curve AUC0-t (μ g/ml
*H) value can calculate, the mouse tail vein prodrug is 112.1% in the relative bioavailability of ubenimex, the mouse oral prodrugs is 96.68% in the relative bioavailability of ubenimex, credit is analysed in tail vein injection and the oral administration group by statistics, the AUC0-t of prodrug and ubenimex there are no significant difference (P〉0.05), prodrug and ubenimex bioequivalence are described; Residence time (MRT 0-t) and transformation period (T1/2) in the body of comparative drug, prodrug and ubenimex T1/2 and MRT 0-t have statistical significance (P＜0.05), be that prodrug group T1/2 and MRT 0-t all have increase than the ubenimex group, after prodrug is made in this explanation, owing to improved water-soluble, make medicine have suitable profit partition ratio, it is long to keep the effective plasma concentration time, can prolong the transformation period of ubenimex.
The main statistical moment parameter of table 6. mouse tail vein injection administration ubenimex and prodrug
Main statistical moment parameter behind table 7. mouse oral administration ubenimex and the prodrug
5. inside and outside stability experiment:
Ubenimex enlightening promise ester is stable in the HCl of water, 0.1mol/L solution; In the PBS of pH7.4 solution, degrade, mainly contain two degraded products ,-individually can be defined as ubenimex according to retention time, in addition-individual be N ' N dimethylethanolamine.
Table 8 vitro stability experiment (pH=7.4PBS)
| Time | Ubenimex enlightening promise ester (degradation amount) | Ubenimex (peak area/concentration) | N ' N dimethylethanolamine (peak area) |
| 0min | 0 | 277821(0.91mg/ml) | 110856 |
| 15min | 18.97％ | 351148(1.15mg/ml) | 265968 |
| 30min | 33.49％ | 414490(1.36mg/ml) | 342049 |
| 1h | 65.21％ | 511193(1.67mg/ml) | 458371 |
| 2h | 79.5％ | 571432(1.87mg/ml) | 448440 |
| 4h | 91.3％ | 758511(2.48mg/ml) | 592127 |
| 8h | 96.92％ | 913148(2.99mg/ml) | 447457 |
| 12h | 98.62％ | 1011371(3.31mg/ml) | 288689 |
| 24h | 98.86％ | 1195366(3.91mg/ml) | 126849 |
From showing various data as can be seen,, play antitumor action because pH value of human body about 7.2-7.3, also can slowly be degraded in vivo so can predict the ubenimex prodrug, thereby discharge the parent drug ubenimex.Even be not degraded, because above-mentioned activity experiment proves that ubenimex prodrug activity is better than the parent drug ubenimex, so ubenimex prodrug itself has good antineoplastic activity too.
The application of aforesaid ubenimex enlightening promise ester in preparation medicine for treating tumor thing.
The preparation purposes
Preferably, ubenimex enlightening promise ester is used to prepare injection.Also can be prepared into and be suitable for oral tablet or dripping pill.
Ubenimex suppresses-all be used for oral administration since straight, and for the cancer patients, particularly Advanced Carcinoma Patient is unfavorable for oral administration, and the bioavailability of oral administration is low, document shows that ubenimex is to discharge in body with the form of former medicine more than 85%.And ubenimex enlightening promise ester belongs to two molecule hydrochlorides, water-soluble height, thereby can make injection, as chloride injection agent, glucose injection agent or the like, can improve the bioavailability of parent drug so greatly, and because prodrug has slow-release function, thereby the parent drug residence time in vivo can be prolonged, improve drug effect, reduce administration number of times.
Description of drawings
Fig. 1 is the active figure of ubenimex enlightening promise ester of the present invention, and X-coordinate is a ubenimex enlightening promise ester concentration (mg/ml), and ordinate zou is tumour inhibiting rate (%).
Fig. 2 is the active comparison diagram of ubenimex enlightening promise ester of the present invention and ubenimex, and X-coordinate is inhibiting rate (%), and prodrug and the ubenimex inhibiting rate under 50mg/ml concentration is respectively 35% and 3%.
Fig. 3 is the mouse tail vein injection ubenimex curve of average medicine-time
Fig. 4 is the mouse prodrug tail vein injection ubenimex curve of average medicine-time
Fig. 5 is the average medicine-time curve behind the mouse oral administration ubenimex
Fig. 6 is average medicine-time curve of mouse oral administration prodrug group ubenimex
Embodiment
The present invention will be further described below in conjunction with embodiment, but be not limited thereto.
Embodiment 1:
Antitumor drug with following array structure, ubenimex enlightening promise ester:
The route of synthetic ubenimex enlightening promise ester:
Ubenimex enlightening promise ester synthetic method, step is as follows:
(1) ubenimex with optical purity is a starting raw material, under the alkaline condition of pH=9-10, makes ubenimex and tert-Butyl dicarbonate reaction (the tert-Butyl dicarbonate molar weight is 1.1 times of ubenimex), and the primary amine groups of ubenimex is carried out the Boc protection,
(2) make solvent with anhydrous tetrahydro furan; under the katalysis of 0 ℃ of temperature and dicyclohexylcarbodiimide (DCC) (the DCC molar weight is 1.1 times of ubenimex); the ubenimex and the HOBt (molar weight is 1.08 times of ubenimex) of the product B oc of step (1) protection are condensed into active ester
(3) going up step reflection its lytic activity ester at room temperature reaches under the katalysis to dimethylamino pyridine (DMAP) (the DMAP molar weight is 0.2 times of ubenimex) again; be reacted into ester with N ' N dimethylethanolamine; generate the prodrug of Boc protection, the mol ratio of described active ester and N ' N dimethylethanolamine is 1:3.
The prodrug of the Boc protection that (4) step (3) is obtained is taken off the Boc blocking group in the saturated ethyl acetate solution of hydrogenchloride, get target compound.
Embodiment 2:
Take by weighing sodium-chlor by the recipe quantity precision, add the water for injection dissolving of recipe quantity 70%, by volume add 1% gac, stir, filter, HCl regulates pH5, adds the ubenimex enlightening promise ester of recipe quantity embodiment 1, stirring and dissolving, water for injection is to 1000ml, filter, embedding is sterilized 30 minutes promptly in the 2ml ampoule.
Embodiment 3:
Take by weighing glucose by the recipe quantity precision, add the water for injection dissolving of recipe quantity 70%, by volume add 1% gac, stir, filter, HCl regulates pH5, adds the ubenimex enlightening promise ester of recipe quantity embodiment 1, stirring and dissolving, water for injection is to 1000ml, filter, embedding is sterilized 30 minutes promptly in the 2ml ampoule.
Embodiment 4:
Take by weighing the ubenimex enlightening promise ester of embodiment 1 by the recipe quantity precision, in medicine: PEG6000: water: glycerine was in 1: 1.8: 0.4: 0.4 ratio mixing, being incubated in the system of the dripping device of (90 ± 2) ℃, is that the dropper of 2.5/3.5 (mm) splashes in the cooling fluid that temperature is a room temperature so that dripping of (10 ± 2) D/min is fast with the drip inner/outer diameter.
Embodiment 4:
Take by weighing the ubenimex enlightening promise ester of embodiment 1 by the recipe quantity precision, add the Microcrystalline Cellulose thorough mixing in 20% ratio, add 5% cross-linked polyvinylpyrrolidone and 0.3% micropowder silica gel then, carry out direct powder compression behind the mixing, the hardness of control tablet is 3～4kgcm-2.Made dispersible tablet can be in 3min disintegration and all sieving fully by No. 2, meet that " Chinese pharmacopoeia is to the requirement of burst.
Embodiment 5:
Take by weighing the ubenimex enlightening promise ester of embodiment 1 by the recipe quantity precision, with HPMC, the IV acrylic resin prepares the sheet heart as main binder, and the hard-hearted degree of sheet reaches more than the 0.5MPa, about disintegration 10min, friability＜1% is unilateral level and smooth.Film-coated optimum process parameter: the Opadry consumption of dressing is 5%, discharge rate 400g/min, rotating speed 15r/min.
Claims (8)
1. the antitumor drug that has following array structure, ubenimex enlightening promise ester:
2. the preparation method of ubenimex enlightening promise ester as claimed in claim 1 is characterized in that this method is: introduce this natural radioactivity small molecules of N ' N dimethylethanolamine on the carboxyl, its structural formula is as follows:
The route of synthetic ubenimex enlightening promise ester:
The synthetic method of ubenimex enlightening promise ester of the present invention, step is as follows:
(1) ubenimex with optical purity is a starting raw material, under the alkaline condition of pH=9-10, makes the reaction of ubenimex and tert-Butyl dicarbonate, and the primary amine groups of ubenimex is carried out the Boc protection,
(2) make solvent with anhydrous tetrahydro furan, under the katalysis of 0 ℃ of temperature and DCC, the ubenimex and the N-hydroxy benzo tetrazole of the product B oc of step (1) protection be condensed into active ester,
(3) go up step reaction product active ester and at room temperature reach under the katalysis of DMAP again, be reacted into ester with N ' N dimethylethanolamine, generate the prodrug of Boc protection, the mol ratio of described active ester and N ' N dimethylethanolamine is 1: 3;
The prodrug of the Boc protection that (4) step (3) is obtained is taken off the Boc blocking group in the saturated ethyl acetate solution of hydrogenchloride, get target compound.
3. the preparation method of ubenimex enlightening promise ester as claimed in claim 2 is characterized in that in the step (1), and ubenimex is 1:1.1 with the molar weight ratio of tert-Butyl dicarbonate.
4. the preparation method of ubenimex enlightening promise ester as claimed in claim 2 is characterized in that in the step (2), and N-hydroxy benzo tetrazole is 1:1.08 with the molar weight ratio of ubenimex, and dicyclohexylcarbodiimide is 1:1.1 with the molar weight ratio of ubenimex.
5. the preparation method of ubenimex enlightening promise ester as claimed in claim 2 is characterized in that in the step (3), is 0.2 times of ubenimex molar weight to the dimethylamino pyridine catalytic amount.
6. the preparation method of ubenimex enlightening promise ester as claimed in claim 2 is characterized in that this method is applicable to that other contains the preparation of the ester class prodrug of carboxyl medicine.
7. one kind has broad applicability, does not influence the one-tenth ester method of intramolecularly active group and reaction conditions gentleness, and this becomes ester method reaction expression as follows:
R1, R2 are various types of substituting groups, and fat group or aromatic group are chain or cyclic substituents.
8. the application of ubenimex enlightening promise ester as claimed in claim 1 in preparation medicine for treating tumor thing.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101665449B (en) * | 2009-09-24 | 2012-12-12 | 山东大学 | Water-soluble prodrug of tamibarotene, and preparation method and applications thereof |
| WO2014194848A1 (en) * | 2013-06-07 | 2014-12-11 | 潍坊博创国际生物医药研究院 | Multi-targeted ubenimex prodrug derivative and preparation method and use thereof |
| CN108977498A (en) * | 2018-06-19 | 2018-12-11 | 潍坊医学院 | A kind of the Inhibiting enzyme activity measuring method and application of aminopeptidase N inhibitor |
| CN109646429A (en) * | 2018-12-21 | 2019-04-19 | 江西润泽药业有限公司 | A kind of solid dosage forms of prodrug derivant and preparation method thereof containing ubenimex |
| CN109846873A (en) * | 2018-12-18 | 2019-06-07 | 江西润泽药业有限公司 | A kind of antineoplastic pharmaceutical compositions |
| CN113876784A (en) * | 2021-09-27 | 2022-01-04 | 潍坊博创国际生物医药研究院 | Novel application of boroleucine compound |
| CN114349816A (en) * | 2021-11-30 | 2022-04-15 | 青岛博创生物科学研究院 | Small molecule coupling molecule based on aminopeptidase N/CD13 and preparation method and application thereof |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4189604A (en) * | 1975-07-22 | 1980-02-19 | Zaidan Hojin Biseibutsu Kagaku Kenkyu Kai | Bestatin |
| WO2004014879A1 (en) * | 2002-08-08 | 2004-02-19 | Fujisawa Pharmaceutical Co., Ltd. | New process |
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- 2009-04-16 CN CN2009100206554A patent/CN101531613B/en not_active Expired - Fee Related
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101665449B (en) * | 2009-09-24 | 2012-12-12 | 山东大学 | Water-soluble prodrug of tamibarotene, and preparation method and applications thereof |
| WO2014194848A1 (en) * | 2013-06-07 | 2014-12-11 | 潍坊博创国际生物医药研究院 | Multi-targeted ubenimex prodrug derivative and preparation method and use thereof |
| JP2016527192A (en) * | 2013-06-07 | 2016-09-08 | ジーナン プラチナム ファーマテック カンパニー リミテッド | Multi-target ubenimex prodrug derivatives and their preparation and use |
| US9861703B2 (en) | 2013-06-07 | 2018-01-09 | Jinan Platinum Pharmatech Co. Ltd. | Multi-targeted Ubenimex prodrug derivative and preparation method and use thereof |
| CN108977498A (en) * | 2018-06-19 | 2018-12-11 | 潍坊医学院 | A kind of the Inhibiting enzyme activity measuring method and application of aminopeptidase N inhibitor |
| CN108977498B (en) * | 2018-06-19 | 2022-09-20 | 潍坊医学院 | Method for determining enzyme inhibition activity of aminopeptidase N inhibitor and application thereof |
| CN109846873A (en) * | 2018-12-18 | 2019-06-07 | 江西润泽药业有限公司 | A kind of antineoplastic pharmaceutical compositions |
| CN109646429A (en) * | 2018-12-21 | 2019-04-19 | 江西润泽药业有限公司 | A kind of solid dosage forms of prodrug derivant and preparation method thereof containing ubenimex |
| CN113876784A (en) * | 2021-09-27 | 2022-01-04 | 潍坊博创国际生物医药研究院 | Novel application of boroleucine compound |
| CN113876784B (en) * | 2021-09-27 | 2023-07-21 | 潍坊博创国际生物医药研究院 | Novel application of boro-leucine compound |
| CN114349816A (en) * | 2021-11-30 | 2022-04-15 | 青岛博创生物科学研究院 | Small molecule coupling molecule based on aminopeptidase N/CD13 and preparation method and application thereof |
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| CN101531613B (en) | 2012-01-18 |
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