CN106916177B - A kind of deuterated dipeptide boronic acid or its ester type compound and its synthetic method and purposes - Google Patents

Abstract

The invention discloses a kind of deuterated dipeptide boronic acid or boric acid ester compound or its crystal form or pharmaceutically acceptable hydrates or solvate, shown in structure such as following formula (a),Wherein: R₁、R₂、R₃、R₄、R₅、R₆Independently selected from hydrogen, deuterium or halogen, or one or more deuterated or complete deuterated C₁‑C₄Alkyl；And R₁、R₂、R₃、R₄、R₅、R₆In at least one be deuterated or deuterium.The compounds of this invention can effective protease inhibition body, effectively treat or prevent cancer, cardiovascular disease, inflammation, immunity disease, nephrosis, angiogenesis or prostatic disorders.

Description

A kind of deuterated dipeptide boronic acid or its ester type compound and its synthetic method and purposes

Technical field

The invention belongs to pharmaceutical synthesis fields, and in particular to a kind of deuterated dipeptide boronic acid or its ester type compound and its conjunction At method and purposes.

Background technique

Ubiquitin-Proteasome Pathway (Ubiquitin-Proteasome Pathway, abbreviation UPP) is intracellular protein The main path of system degradation, and many physiologically important cellular processes are participated in, including signal transduction, immune response are not rolled over Folded albumen reaction and cell cycle progress.The hair of this approach and cardiovascular and cerebrovascular disease, cancer and nervous system degenerative disease Disease etc. suffers from important relationship.For this novel target spot, first proteasome inhibitor bortezomib (PS- in 2003 341) FDA approval listing is obtained, for treating recurrent myeloma.The medicine in 2004 goes through to list in European Union again, for multiple Property myeloma.In September, 2005, the medicine are introduced by Xi'an Yang Sen, are listed for the first time in China Guangzhou.In on July 11st, 2007 and quilt U.S. FDA is ratified for treating relapsed or stubborn lymphoma mantle cell (Mantle Cell Lymphoma, abbreviation MCL). The sales volume of Bortezomib in 2014 reaches 30.69 hundred million dollars, becomes one of 20 before the best-selling tumour medicine in the whole world.Bortezomib is in China Market price is every 3.5 milligrams about 13000 yuan, treats about 40,000 yuan of a cycle expense, so expensive expense is for many trouble Person is very heavy financial burden.In December, 2015, U.S. FDA had approved first oral protein enzyme body inhibitor again Ixazomib listing, for treating Huppert's disease.But current clinical data is shown, there is also relatively more for this kind of drug Side effect, such as tired out of strength, nauseous, diarrhea and neuropathy.Moreover, therefore, how to develop it is a kind of it is cheap, The low high therapeutic protein enzyme body inhibitor oral drugs of toxic side effect are that we need key problems-solving at present.

Summary of the invention

Goal of the invention: in view of the above technical problems, the object of the invention offer is a kind of novel to live with proteasome inhibition The compound and its preparation method and use of property and more preferable pharmacodynamics performance.

Technical solution: first aspect present invention disclose a kind of deuterated dipeptide boronic acid or boric acid ester compound or its Crystal form or pharmaceutically acceptable hydrate or solvate, shown in structure such as following formula (a),

Wherein:

R₁、R₂、R₃、R₄、R₅、R₆Independently selected from hydrogen, deuterium or halogen, or one or more deuterated or complete deuterated C₁-C₄ Alkyl；And R₁、R₂、R₃、R₄、R₅、R₆In at least one be deuterated or deuterium；

P group is selected from following segment:

Wherein, B is boron atom, Z₁、Z₂Independently selected from hydroxyl, C_1-10Alkoxy or aryloxy group；

Or P group is to be formed by the heterocyclic group containing N, S and/or O by following segment and other compounds,

Wherein B is boron atom, and the atom connecting with formula (a) is B.

It is preferred that the R₁And R₄It is halogen, R₂、R₃、R₅、R₆In there are two hydrogen and two deuteriums.

It is preferred that the R₁And R₄It is chlorine, R₂And R₃It is hydrogen, R₅And R₆It is deuterium.

It is preferred that P group is to form borate or anhydride group by following segment and alpha-hydroxy carboxylic acid compounds or beta-hydroxycarboxylic acids,

Wherein B is boron atom, and the atom connecting with formula (a) is B.

It is preferred that P group is that by following segment and have the not independent hydroxyl of two or more on same carbon atom The compound of structure forms borate group,

Wherein B is boron atom, and the atom connecting with formula (a) is B.

In another preferred example, the deuterated deuterium isotopic content in deuterium the position of substitution is at least greater than natural deuterium isotope and contains It measures (0.015%), is preferably greater than 30%, even more preferably greater than 50%, even more preferably greater than 75%, even more preferably greater than 95%, more preferably Ground is greater than 99%.

In another preferred example, formula (I) compound at least contains 1 D-atom, more preferably 2 D-atoms.

In another preferred example, R₁、R₄Selected from halogen；More preferably chlorine；

In another preferred example, R₂、R₃For hydrogen；

In another preferred example, R₅、R₆Separately it is selected from hydrogen, deuterium, deuterated methyl, deuterated ethyl, halogen.

Borinate ester moiety is five-membered ring in some embodiments.In further embodiments, borinate ester moiety is hexatomic ring. In some other embodiment, borinate ester moiety is the mixture of five-membered ring and hexatomic ring.

As used herein, term " independently selected from " is that multiple groups are respectively selected from certain substituent groups, and each group it Between do not connect each other, such as " R₁、R₂Independently selected from hydrogen, deuterium or halogen ", refer to R₁Selected from hydrogen, deuterium or halogen, R₂It is selected from Hydrogen, deuterium or halogen, and R₁And R₂It is not connected each other between group.

As used herein, term " alpha-hydroxy carboxylic acid compounds " refers to containing the alpha position being directly connected to relative to carboxylic acid group The compound of hydroxyl on carbon atom.As used herein, term " alpha-hydroxy carboxylic acid compounds " be not limited to only to have hydroxyl and The compound of one carboxylic acid group.

As used herein, term " beta-hydroxycarboxylic acids " refers to containing the carbon for being directly connected to the position β relative to carboxylic acid group The compound of hydroxyl on atom.As used herein, term " beta-hydroxycarboxylic acids " is not limited to only have a hydroxyl and one The compound of a carboxylic acid group.

In some embodiments, beta-hydroxycarboxylic acids are selected from malic acid, citric acid, 3-hydroxybutyrate, beta-hydroxy isovaleric acid And salicylic acid.In some of the other embodiments, beta-hydroxycarboxylic acids are selected from malic acid, and citric acid, 3-hydroxybutyrate, beta-hydroxy is different Valeric acid, tartaric acid and salicylic acid.In certain embodiments, beta-hydroxycarboxylic acids are citric acids.Beta-hydroxycarboxylic acids it is some other Non-limiting example includes glucoheptonic acid, maltobionic acid, lactobionic acid and galactosaccharic acid.Some other non-limits of beta-hydroxycarboxylic acids Property example processed includes enemy's oxalic acid, 1- hydroxy-2-naphthoic acid and 3- hydroxy-2-naphthoic acid.

In some embodiments, 'alpha '-hydroxy acids or beta-hydroxy acid are selected from glycolic, malic acid, hexahydro mandelic acid, 2- hydroxyl Base isobutyric acid, citric acid, mandelic acid, lactic acid, 3- hydroxy-butyric acid, beta-hydroxy isovaleric acid, 2- hydroxyl -3,3- acid dimethyl, 2- Hydroxy-3-methyl butyric acid ,-hydroxy isocaproic acid, tartaric acid, salicylic acid and benzoic acid.

In some embodiments, there is the change of the not independent hydroxyl structure of two or more on same carbon atom It closes object and is selected from diethanol amine, tartaric acid, alpha-D-glucose, D-ribose etc..

When above-mentioned alpha-hydroxy carboxylic acid compounds or beta-hydroxycarboxylic acids are citric acids, the structure of formula (a) can be following two structure:

In other embodiments, when alpha-hydroxy carboxylic acid compounds or beta-hydroxycarboxylic acids are citric acids, between carboxylic acid and boron atom Other key can be formed.It is not limited by any chemical bonding theory, in such embodiments, formula (a) can also be by Formula (3) or (4) indicate:

It is not limited by any chemical bond and theory there is also other, can be used for describing such as (3) or (4) hydroxycarboxylic acid With the further key of boric acid and other forms, no longer describe one by one herein.

R in the compounds of this invention₅、R₆The carbon atom that group is connected can be raceme, it is possible to have optics is living Property.

In another preference, said compound is preferred compound selected from the group below；

((R) -1- (two deuterium acetylamino of 2- (2,5- benzamido) -2,2-) -3- methyl butyl) boric acid；

((R) -1- (two deuterium acetylamino of 2- (2,5- benzamido) -2,2-) -3- methyl butyl) boric acid-(+)-australene Alkane diol ester

((R) -1- (two deuterium acetylamino of 2- (2,5- benzamido) -2,2-) -3- methyl butyl) borocitric acid ester

((R) -1- (the deuterated dimethyl-acetylamino of 2- (2,5- benzamido) -2,2-) -3- methyl butyl) boric acid Chinese holly Rafter acid esters

((R) -1- (two deuterium acetylamino of 2- (2,5- benzamido) -2,2-) -3- methyl butyl) borate tartar acid esters

Tartaric acid ((R) -1- (two deuterium acetylamino of 2- (2,5- benzamido) -2,2-) -3- methyl butyl) borate

((R) -1- (two deuterium acetylamino of 2- (2,5- benzamido) -2,2-) -3- methyl butyl) boric acid alpha-hydroxypentyl Two acid esters

((R) -1- ((S) 2- (2,5- benzamido) -2- deuterium-acetylamino) -3- methyl butyl) boric acid malate

((R) -1- (two deuterium acetylamino of 2- (2,5- benzamido) -2,2-) -3- methyl butyl) boric acid diethanol amine Ester

((R) -1- ((R) 2- (2,5- benzamido) two deuteriums of -2,2--acetylamino) -3- methyl butyl) boric acid grape Sugar ester

Two Portugal of ((R) -1- ((R) 2- (2,5- benzamido) two deuteriums of -2,2--acetylamino) -3- methyl butyl) boric acid Grape sugar ester

((R) -1- ((R) 2- (2,5- benzamido) two deuteriums of -2--acetylamino) -3- methyl butyl) boric acid ribose ester

Second aspect of the present invention provide the deuterated dipeptide boronic acid or boric acid ester compound or its crystal form or The preparation method of pharmaceutically acceptable hydrate or solvate, including reaction process as shown below:

Wherein, P group is to be formed by the heterocyclic group containing N, S and/or O by following segment and other compounds,

Wherein B is boron atom, and the atom connecting with formula (a) is B.

A kind of particularly preferred compound of the invention is ((R) -1- (two deuterium acetyl of 2- (2,5- benzamido) -2,2- Amino) -3- methyl butyl) boric acid diethanol amine ester (compound N NU-458).

The particularly preferred compound of another kind of the invention is ((R) -1- (two deuterium second of 2- (2,5- benzamido) -2,2- Acylamino-) -3- methyl butyl) borocitric acid ester

The present invention provides their preparation methods.

The described method includes:

In the ethyl acetate of boiling, compound (VIII) i.e. NNU-455 is reacted with diethanol amine (DEA), by slowly dropping Warm crystallization obtains final product NNU-458；

In another preferred example, NNU-455 can be allowed to react with citric acid by similar approach, generates NNU-459.

In another preferred example, compound (VIII) is prepared by the following method:

In atent solvent, heavy water, DMSO-d₆And under the action of alkali, compound (VII) reaction forms the compound (Ⅷ)。

In another preferred example, compound (VII) is prepared by the following method:

Compound (VI) is reacted with isobutaneboronic acid under the action of an acid generates compound (VII)；

In another preferred example, compound (VI) is prepared by the following method:

In atent solvent, compound (IV) and compound (V) are under the action of condensing agent and alkali, dehydrating condensation shape At the compound (VI).

In another preferred example, compound (IV) obtains by the following method:

In the solution of first alcohol and water, compound (III) demethylation obtains compound (IV) in the presence of alkali

In another preferred example, compound (III) obtains by the following method:

In atent solvent, 2,5- dichlorobenzoic acids (I) and compound (II) are changed under the action of condensing agent and alkali It closes object (III)

In another preferred example, the alkali is selected from potassium carbonate, cesium carbonate, sodium hydride, hydrofining, potassium hydroxide, hydrogen-oxygen Change sodium or combination.

Common peptide condensing agent is N in above-mentioned reaction, and N- dicyclohexyl-carbodiimide (is abbreviated as DCC), 1- (3- diformazan Aminopropyl) -3- ethyl-carbodiimide hydrochloride (being abbreviated as EDC.HCl), 1- hydroxy benzo triazole (being abbreviated as HOBt) or Isobutyl chlorocarbonate.

In another preferred example, each reaction temperature carries out at -20~100 DEG C.

In the third aspect of the present invention, a kind of method for preparing pharmaceutical composition is provided, comprising steps of will pharmaceutically may be used Compound described in the carrier and first aspect present invention of receiving or its pharmaceutically acceptable crystal form, hydrate or solvent It closes object to be mixed, to form pharmaceutical composition.

In the fourth aspect of the present invention, a kind of pharmaceutical composition is provided, it contains pharmaceutically acceptable carrier and this Compound described in invention first aspect or its crystal form, pharmaceutically acceptable hydrate or solvate.

In another preferred example, the pharmaceutical composition is injection, wafer, tablet, pill, powder or granule.

In another preferred example, the pharmaceutical composition also contains other therapeutic agent, the other treatment Drug is the drug of cancer, cardiovascular disease, inflammation, immunity disease, nephrosis, angiogenesis, prostatic disorders.

More preferably, the therapeutic agent includes (but being not limited to): 5 FU 5 fluorouracil, AV412, Avastin TM (avastin, bevacizumab), bexarotene (bexarotene), bortezomib (bortezomib), calcitriol (calcitriol), Canertinib (canertinib), capecitabine (capecitabine), Carboplatin (carboplatin), plug To examine former times (celecoxib), Cetuximab (cetuximab), CHR-2797, cis-platinum (cisplatin), Dasatinib (dasatinib), digoxin (digoxin), enzastaurin, Erlotinib (Erlotinib) rely on pool examination (etoposide), everolimus (everolimus), fulvestrant (fulvestrant), Gefitinib (gefitinib), 2, 2- difluoro deoxidation born of the same parents crash smack one's lips core former times (gemcitabine), Genistein (genistein), Imatinib (imatinib), according to It is vertical to replace health (irinotecan), Lapatinib (lapatinib), lenalidomide (lenalidomide), come bent mile (letrozole), folinic acid (leucovorin), matuzumab ((matuzumab), oxaliplatin (oxaliplatin), Taxol (paclitaxel), Victibix (panitumumab), PEGylated granulocyte colony stimulating factor (pegfilgrastin), PEGylated alpha-interferon (peglated alfa-interferon), training U.S. are bent (pemetrexed), Polyphenon E, Satraplatin (satraplatin), sirolimus (sirolimus), Sutent (sutent, Sunitinib), sulindac (sulindac), taxotere (taxotere), for not mile amine (temodar, Temozomolomide), Torisel (temsirolimus), TGOltipifarnib, trastuzumab (trastuzumab), third Valeric acid (valproic acid), vinflunine (vinflunine), Volociximab, vorinostat and XL647.

In the fifth aspect of the invention, compound described in first aspect present invention or its crystal form, pharmaceutically are provided The purposes of acceptable salt, hydrate or solvate, they be used to prepare the pharmaceutical composition of protease inhibition body.

In another preferred example, the compound can be used for treating and preventing disease relevant to proteasome target spot Disease.

In another preferred example, the pharmaceutical composition is for treating and preventing following disease: cancer, cardiovascular disease Disease, inflammation, immunity disease, nephrosis, angiogenesis or prostatic disorders.

In another preferred example, the cancer include (but being not limited to): Huppert's disease, non-small cell lung cancer, Uterine cancer, rectum, the cancer of the brain, head cancer, neck cancer, cutaneum carcinoma, prostate cancer, breast cancer, entity tumor, kidney, leukemia, liver cancer, stomach Cancer or cancer of pancreas.

In the sixth aspect of the present invention, a kind for the treatment of method is provided, it to object in need for the treatment of comprising steps of apply Compound described in first aspect present invention or its crystal form, pharmaceutically acceptable hydrate or solvate, or application is originally Pharmaceutical composition described in the invention third aspect, thus protease inhibition body.Preferably, the disease includes: cancer, the heart Vascular diseases, inflammation, immunity disease, nephrosis, angiogenesis or prostatic disorders.

It should be understood that above-mentioned technical characteristic and each technical characteristic hereinafter specifically described can in the scope of the invention To be combined with each other, to form a new or preferred technical solution.Due to space limitations, I will not repeat them here.

Specific embodiment

It has been investigated that deuterated dipeptide boronic acid and borate of the invention compared with not deuterated compound, have more excellent Different pharmacokinetics and/or pharmacodynamics performance, therefore it is more suitable for the compound of protease inhibition body, and then be more suitable for Prepare the drug for the treatment of cancer and related disease.The present invention is completed on this basis.

With ((R) -1- (two deuterium acetylamino of 2- (2,5- benzamido) -2,2-) -3- methyl butyl) borocitric acid For ester (compound N NU-459).

Pharmacokinetics in Rat experimental result shows, the half-life period T of NNU-459 ratio MLN9708_2/1Extend, under curve Area AUC is dramatically increased, and the apparent clearance rate of NNU-459 is also substantially reduced than MLN9708.Simultaneously in the hepatomicrosome stability of people In experiment, the half-life period (T of the activity in vivo ingredient (NNU-455) of compound N NU-459_1/2=108.3min) also than Half-life period (the T of the active constituent (MLN2238) of MLN9708_1/2=57.3min) significantly increase.

Furthermore in vitro experiment, to tumor cell line U266, RPMI-8266 and breast cancer cell line MAD-MB-231 The IC of inhibition₅₀(nM) on, NNU-459 also shows better activity than control compound MLN-9708.

Definition

As used herein, " deuterated " refers to that one or more hydrogen in compound or group are replaced by deuterium.It is deuterated to can be one Replace, two replace, polysubstituted or full substitution.Term " one or more deuterated " with " one or many deuterated " is interchangeable makes With.

In another preferred example, deuterated to replace unknown deuterium isotopic content to be at least greater than natural deuterium isotope and contain in deuterium It measures (0.015%), is preferably greater than 30%, be more preferably greater than 50%, even more preferably greater than 75%, even more preferably greater than 95%, more preferably Ground is greater than 99%.

In another preferred example, formula compound at least contains 1 D-atom, more preferably 2 D-atoms.

Active constituent

Term " the compounds of this invention " as used herein refers to compound shown in formula (a).The term further includes and formula (a) chemical combination Various crystalline forms, pharmaceutically acceptable hydrate or the solvate of object.

Preparation method

The preparation method of formula (a) structural compounds is described more particularly below, but these specific methods are not to this Invention constitutes any restrictions.The compounds of this invention can also optionally will be describing or known in the art various in the present specification Synthetic method combines and is easily made, such combination can by those skilled in the art in the invention readily into Row.

The preparation method for the not deuterated dipeptide boronic acid and its esters that the present invention uses is known.Corresponding deuterated dipeptides Boric acid and its esters can be raw material with corresponding deuterated boronic acid compounds.It is synthesized with same route.

By taking NNU-459 as an example, a kind of preferred preparation flow is as follows.

Synthetic route:

2,5- dichlorobenzoic acids (chemical compounds I) and glycine methyl ester hydrochloride (compound ii), in peptide condensing agent and alkali The lower reaction of effect obtains (2,5- dichloro-benzoyl base) glycine methyl ester (compound III)；Then compound III is in the presence of alkali Saponification occurs and obtains (2,5- dichloro-benzoyl base) glycine (compounds Ⅳ)；Compounds Ⅳ with (aR, 3aS, 4S, 6S, 7aR)-hexahydro -3a, 8,8- trimethyl-alpha- (2- methyl-propyl) -4,6- first bridge -1,3,2- benzo dioxy borine -2- methylamine For 2,2,2- trifluoroacetates (compound V) under the action of peptide condensing agent and alkali, reaction generates ((R) -1- (2- (2,5- benzene first Amide groups) two deuterium acetylamino of -2,2-) -3- methyl butyl) boric acid-(+)-australene alkane diol ester (compound VI)；In methanol In, compound 5 reacts generation ((R) -1- (2- (2,5- benzamido)-acetyl with isobutaneboronic acid under the action of 1N hydrochloric acid Amino) -3- methyl butyl) boric acid (compound VII)；Then in tetrahydrofuran, in the work of potassium carbonate and deuterated DMSO and heavy water Under, ((R) -1- (bis- deuterium acetylamino of 2- (2,5- benzamido) -2,2-) -3- methyl butyl) boric acid (compound is generated Ⅷ)；Compound VII reacts to obtain compound N NU-459 with citric acid in the ethyl acetate of boiling.

Specific synthetic method is discussed in detail in embodiment 1.

Pharmaceutical composition and method of administration.

Since the compounds of this invention has an excellent inhibitory activity to proteasome, the compounds of this invention and its each Crystal form, pharmaceutically acceptable hydrate or solvate are planted, and is the medicine of main active containing the compounds of this invention Compositions can be used for treating, prevent and alleviate the disease by mediating to proteasome.According to the prior art, chemical combination of the present invention Object can be used for treating following disease: cancer, cardiovascular disease, obesity, diabetes etc..

Pharmaceutical composition of the invention include safe and effective amount within the scope of the compounds of this invention or its be pharmacologically subjected to Salt and pharmacologically acceptable excipient or carrier.Wherein " safe and effective amount " refers to: the amount of compound is enough obviously Improve the state of an illness, and is unlikely to generate serious side effect.In general, pharmaceutical composition contains 1-2000mg the compounds of this invention/agent, More preferably, containing 10-200mg the compounds of this invention/agent.Preferably, described is " one " for a capsule or tablet.

" pharmaceutically acceptable carrier " refers to: one or more biocompatible solids or liquid filler or jello Matter, they are suitable for people's use and it is necessary to have enough purity and sufficiently low toxicity." compatibility " referred to herein as combines In object each component energy and the compound of the present invention and they between mutually admix, and significantly reduce the drug effect of compound.

Term " carrier ", " excipient " or " carrier " are used interchangeably herein, and including any and all solvents, Diluent and other liquid-carriers, dispersion or suspension aids, surfactant, pH adjusting agent, isotonic agent, thickening or emulsification examination Agent, preservative, solid binder, lubricant etc. are suitable for required particular dosage form.Unless any conventional carrier medium and this hair Bright compound is incompatible, for example, by generate any undesirable biological effect or with any other group of split-phase of pharmaceutical composition Interaction makes compound reduction or failure, or generates harmful substance, and otherwise its purposes is considered in the scope of the present invention.

Pharmaceutically acceptable carrier includes but is not limited to ion-exchanger, aluminium oxide, aluminum stearate, lecithin, blood Albumin such as human serum albumins, buffer substance such as phosphate, carbonate, magnesium hydroxide and aluminium glycine, sorbic acid or sorb Sour potassium, the partial glyceride mixture of saturated vegetable fatty acid, water, apirogen water, salt or electrolyte such as protamine sulfate, phosphoric acid Disodium, disodium hydrogen phosphate, sodium chloride and zinc salt, colloidal silicon dioxide, magnesium trisilicate, polyvinylpyrrolidone, polyacrylate, Polyethylene-polyoxypropylene block polymer, lanolin, sugar such as lactose, glucose, sucrose and mannitol, starch such as cornstarch And potato starch, cellulose and its derivates such as sodium carboxymethylcellulose, ethyl cellulose and cellulose acetate, powder Huang alpine yarrow Glue；Malt, gelatin, talcum powder, excipient such as cocoa butter and suppository wax, oil such as peanut oil, cottonseed oil, sunflower oil, sesame oil, Olive oil, corn oil and soybean oil, glycol such as propylene glycol and polyethylene glycol, ester such as ethyl oleate and ethyl laurate, agar, sea Alginic acid, isotonic saline solution, compound sodium chloride injection, alcohol such as ethyl alcohol, isopropanol alcohol, hexadecanol and glycerol, cyclodextrin such as hydroxypropyl Beta-cyclodextrin and sulfobutyl ether beta-cyclodextrin, lubricant such as lauryl sodium sulfate and magnesium stearate, petroleum hydrocarbon such as mineral oil and Vaseline.Colorant, release agent, coating agent, sweetener, flavoring agent and aromatic, preservative and antioxidant can also bases The judgement of formulator is present in composition.

Pharmaceutical composition of the invention can be prepared by methods known in the art, such as conventional granulation, mixing, molten Solution, encapsulating, freeze-drying or emulsification method etc..Composition can produce in a variety of manners, including particle, precipitating or particle, powder, packet Include freeze-drying, rotary drying or spray-dried powders, amorphous powder, tablet, capsule, syrup, suppository, injection, emulsion, Elixir, suspension or solution.

The method of application of the compounds of this invention or pharmaceutical composition is not particularly limited, will according to preferred embodiment Composition of the invention is formulated for mammal, preferably to the medicament administration of the mankind.These pharmaceutical compositions of the invention Object can be by oral, alimentary canal external administration, part spraying by sucking, rectum, nose, cheek, vagina or via implanted reservoir Application.The term as used herein " alimentary canal external administration " is including subcutaneous, intravenously, intramuscular, intra-articular, intrasynovial, in breastbone, sheath It is interior, in liver and intracranial injection or infusion techniques.Preferably, composition is oral, intravenously or subcutaneously applies.Preparation of the invention can It is short-acting to be designed as, quick release or long-acting.In addition, compound can locally rather than systemic fashion is applied, such as in tumour Apply (such as passing through injection) in position.

Liquid formulation for oral administration includes but is not limited to pharmaceutically acceptable emulsion, microemulsion, solution, is mixed Suspension, syrup and elixir.Other than reactive compound, liquid dosage form can contain inert diluents commonly used in the art Agent, such as water or other solvents, solubilizer and emulsifier, such as ethyl alcohol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, Ergol, propylene glycol, 1,3-BDO, cyclodextrin, dimethylformamide, oily (especially cottonseed oil, peanut oil, corn Oil, embryo oil, olive oil, castor oil and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol and aliphatic ester, anhydro sorbitol And its mixture.Besides inert diluents, Orally administered composition may also include adjuvant, such as wetting agent, emulsifier and suspending agent, sweet tea Taste agent, spin agent and aromatic.

Solid dosage forms for oral administration includes capsule, tablet, pill, powder and granule.In these solid formulations In type, reactive compound is mixed at least one conventional inert excipients (or carrier), such as sodium citrate or Dicalcium Phosphate, or with Following compositions mixing: (a) filler or expanding material, for example, starch, lactose, sucrose, glucose, mannitol and silicic acid；(b) it bonds Agent, for example, hydroxymethyl cellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and Arabic gum；(c) moisturizer, example Such as, glycerol；(d) disintegrating agent, for example, agar, calcium carbonate, potato starch or tapioca, alginic acid, certain composition silicates, And sodium carbonate；(e) retarding solvent, such as paraffin；(f) absorbsion accelerator, for example, quaternary ammonium compound；(g) wetting agent, for example, spermaceti Pure and mild glycerin monostearate；(h) adsorbent, for example, kaolin；(i) lubricant, for example, talcum, calcium stearate, stearic acid Or mixtures thereof magnesium, solid polyethylene glycol, lauryl sodium sulfate,.In capsule, tablet and pill, dosage form also may include delaying Electuary.

Coating and shell material preparation can be used in solid dosage forms such as tablet, sugar-pill, capsule, pill and granule, such as casing and Other materials well known in the art.They may include opacifying agent, also, reactive compound or compound in this composition Release can discharge in certain a part in the digestive tract in a delayed fashion.The example of adoptable embedding component is polymeric material And wax material.When necessary, reactive compound can also be with one of above-mentioned excipient or a variety of formation microencapsulation forms.

Other than these inert diluents, composition also may include auxiliary agent, such as wetting agent, emulsifier and suspending agent, sweet taste Agent, corrigent and fragrance.

Injectable formulation, such as sterile injection is aqueous or oily suspensions, can use suitable point according to known technology Powder or wetting agent and suspending agent are prepared.Sterile injectable preparation can also be in parenteral acceptable diluent or solvent In sterile injectable solution, suspension or lotion, such as the solution in 1,3-BDO.What be can be used is acceptable Carrier and solvent be water, compound sodium chloride injection and isotonic sodium chlorrde solution.In addition, sterile expressed oi is usually used Make solvent or suspension media, any mild expressed oi, monoglyceride or diglyceride including synthesis can be used.This Outside, fatty acid such as oleic acid is used to prepare injection.Injectable formulation can for example, by by bacteria retaining filter filtering or It is sterilized by mixing the bactericidal agent of aseptic solid composite form, the aseptic solid composite can use preceding dissolution or divide It dissipates in sterile water or other sterile injectable mediums.The composition for being formulated for parenteral can be by injecting or by fixed When bolus infusion, or can be administered by continuous infusion.

Dosage form for part or transdermal administration the compounds of this invention includes ointment, powder, creme, lotion, gel, powder Agent, solution, spray, inhalant or patch.By active component aseptically with pharmaceutically acceptable carrier and any The preservative needed or the buffer that may be needed mixing.Eye-drops preparations, auristilla and eye drops are recognized as of the invention In range.In addition, the present invention considers to use transdermal patch, have excellent to adding for the controlled delivery of body offer compound Point.Such dosage form can be prepared by the way that compound to be dissolved or dispersed in medium appropriate.Absorption enhancer also can be used The flux of skin is passed through in increase compound.It can be by providing rate controlling membranes or by the way that compound is dispersed in polymer matrix Carry out speed control in matter or gel.

In some embodiments, the present invention provides the pharmaceutical compositions of compound and other excipient as described herein Object.In some of the other embodiments, the present invention provides the medicines of the compound of NNU-455 and other excipient as described herein Compositions.In other embodiments, the present invention provides citric acid esters (NNU-459) and diethanol comprising NNU455 The pharmaceutical composition of amine ester (NNU-458) and other excipient as described herein.

In some embodiments, pharmaceutical preparation of the invention is by using the figuration with low water or low moisture content Agent, and prepared using dry or non-aqueous formulation method, the stable solid oral dosage form of reactive compound is provided.

The compounds of this invention can be administered alone, or be administered in combination with other pharmaceutically acceptable compounds.

It is the mammal that the compounds of this invention of safe and effective amount is applicable to treatment when using pharmaceutical composition (such as people), wherein dosage is the effective dosage pharmaceutically thought when application, for the people of 60kg weight, day is to medicament Amount is usually 1~2000mg, preferably 20~500mg.Certainly, specific dosage is also contemplated that administration route, patient health situation etc. Factor, within the scope of these are all skilled practitioners technical ability.

The compound of the present invention has series of advantages compared with the compound well known in the prior art for not carrying deuterium:

(1) the compounds of this invention has excellent inhibition to proteasome.

(2) metabolism in organism is changed by this technology of deuterate, extends the metabolic half life of drug, this leads to head Cross the reduction of effect (First-pass effect).In such a case, it is possible to change dosage and form durative action preparation, Applicability can be improved in the form of durative action preparation.

(3) pharmacokinetics effect is also changed by deuterate, because deuterated compound forms another hydrate film completely, So that the distribution in organism differs markedly from not deuterated compound.

(4) compound can be improved in animal body due to its isotope effect with the hydrogen atom in deuterium substituted compound Drug concentration, to improve curative effect of medication.

Present invention will be further explained below with reference to specific examples.It should be understood that these embodiments are merely to illustrate the present invention Rather than it limits the scope of the invention.In the following examples, the experimental methods for specific conditions are not specified, usually according to conventional strip Part, or according to the normal condition proposed by manufacturer.Unless otherwise stated, otherwise parts and percentages are parts by weight and weight percent Than.

Embodiment 1:(S) two deuterium acetamide of-N- (2,5- dichloro-benzoyl base) -2,2--D-Leu borocitric acid ester (compound N NU-459).Synthetic route:

(S) preparation of-N- (2,5- dichloro-benzoyl base) glycine (IV):

2,5- dichlorobenzoic acid (7.6g, 40mmol) and HOBt (8.1g, 40mmol) are dissolved in CH₂Cl₂In (200mL), React 10min at -10 DEG C, EDC.HCl (11.5g, 60mmol) be added and reacts 30min, be added compound ii (5g, 40mmol), DIPEA (18.1g, 140mmol) is added after reacting 10min, is warmed to room temperature reaction overnight after reacting 20min.TLC inspection Survey reaction, respectively with 10% hydrochloric acid solution (200mL), 5% NaHCO₃Solution

The washing of (200mL) and saturated salt solution (2 × 200mL), CH₂Cl₂Layer uses anhydrous Na₂SO₄It dries, filters, decompression is steamed Except solvent, oily compound 9.32g is obtained, yield 88.9%.

It takes compound III (129mg, 0.31mmol) obtained by step to be dissolved with 2.5mLMeOH, LiOHH is added₂O (39mg, 0.92mmol) and H₂O (0.8mL), TLC are detected, end of reaction after 2h.It is spin-dried for organic phase, with ether (2 × 1mL) aqueous phase extracted, It is 2~3 that hydrochloric acid to pH value, which is added dropwise, in water phase, generates a large amount of white solids, ethyl acetate extraction, and evaporating solvent under reduced pressure obtains white (S)-N- (2,5- dichloro-benzoyl base) glycine 106mg, yield 86.0%, m.p.169.3-170.8 DEG C.¹H NMR (400MHz,DMSO)δ3.91(-CH₂, d, J=6.0Hz, 2H), 7.48 (- CONH, d, J=8.7Hz, 1H), 7.55 (- Ph, d, J =1.3Hz, 2H), 8.89 (- Ph, t, J=5.9Hz, 1H), 12.71 (- COOH, s, 1H) .MS (ESI): m/z 246.1 [M- H]^-.

(S) system of-N- (2,5- dichloro-benzoyl base) acetamide-D-Leu boric acid-(+)-australene alkane diol ester (VI) It is standby

Compounds Ⅳ (340mg, 0.84mmol) and HOBt (218g, 1.67mmol) are dissolved in CH₂Cl₂In (18mL) ,- 10min is reacted at 10 DEG C, and EDC.HCl (321mg, 1.67mmol) is added and reacts 30min, is added (aR, 3aS, 4S, 6S, 7aR)- Hexahydro -3a, 8,8- trimethyl-alpha- (2- methyl-propyl) -4,6- first bridge -1,3,2- benzo dioxy borine -2- methylamine 2,2, DIPEA (433mg, 3.35mmol) is added after reacting 10min, after reacting 20min in 2- trifluoroacetate (317mg, 0.84mmol) It is warmed to room temperature reaction overnight.TLC detection reaction, respectively with 10% hydrochloric acid solution (20mL), 5% NaHCO₃Solution (20mL) It is washed with saturated salt solution (2 × 20mL), CH₂Cl₂Layer uses anhydrous Na₂SO₄It dries, filters, evaporating solvent under reduced pressure is chromatographed through column Separate to obtain oily target compound 480mg, yield 87.6%.

¹H NMR(400MHz,CDCl₃)δ0.83(-CH₃,s,3H),0.91(-CH₃,s,6H),1.19(-CH₂, d, J= 10.8Hz,1H),1.24(-CH₂, d, J=7.1Hz, 1H), 1.27 (- CH₃,s,3H),1.38(-CH₃,s,3H),1.59-1.69 (-CH,m,1H),1.70(-CH,s,1H),1.77-1.85(-CH₂,m,1H),1.86-1.92(-CH₂,m,1H),1.96-2.01 (-CH,m,1H),2.11-2.21(-CH₂,m,1H),2.25-2.36(-CH₂,m,1H),3.31(-CH,dd,J₁=6.2Hz, J₂ =14.5Hz, 1H), 4.15 (- CH₂, d, J=5.3Hz, 2H), 4.28 (- CH, dt, J₁=6.3Hz, J₂=12.5Hz, 1H), 6.39 (- CONH, d, J=5.1Hz, 1H), 7.24 (- CONH, d, J=4.6Hz, 1H), 7.34 (- Ph, d, J=1.4Hz, 2H), 7.58-7.65(-Ph,m,1H).MS(ESI):m/z 495.3[M+H]⁺.

The preparation of ((R) -1- (2- (2,5- benzamido)-acetylamino) -3- methyl butyl) boric acid (VII):

Compound VI (317mg, 0.49mmol) is dissolved in the MeOH of 3mL, sequentially add isobutaneboronic acid (247mg, 2.43mmol), n-hexane (3mL) and 1N HCl (1.2mL, 1.2mmol), reaction is stirred overnight.TLC detection reaction, n-hexane It is mutually extracted 2 times with MeOH (2 × 3mL), n-hexane (3mL) washs methanol phase 1 time, removes methanol, CH under reduced pressure₂Cl₂(2 × 2mL) extraction Water intaking phase 2 times, washs organic phase to water phase with saturated salt solution (3 × 5mL) and is in neutrality.Evaporating solvent under reduced pressure, through column chromatography point From obtaining pure products 193mg, yield 76.5%.¹H NMR(400MHz,DMSO)δ0.82(-CH₃,s,3H),0.84(-CH₃,s, 3H),1.19-1.28(-CH₂,m,2H),1.61(-CH,td,J₁=6.6Hz, J₂=13.2Hz, 1H), 2.66 (- CH, s, 1H), 4.04(-CH₂, d, J=5.6Hz, 2H), 7.55 (- Ph, d, J=1.3Hz, 2H), 7.66 (- Ph, s, 1H), 8.82 (- CONH, d, J=46.3Hz, 1H), 8.99 (- CONH, t, J=5.7Hz, 1H)¹³C NMR(CDCl₃,100MHz)δ22.91,25.94, 39.90,44.28,60.37,129.20,129.45,131.25,131.34,132.97,135.53,166.38,171.16.MS (ESI):m/z 359.2[M-H]-.HRMS(ESI):calcd for C₁₄H₁₉BCl₂N₂NaO₄[M+Na]+383.0710,found 383.0727.

The system of ((R) -1- (two deuterium acetylamino of 2- (2,5- benzamido) -2,2-) -3- methyl butyl) boric acid (VIII) It is standby:

Compound VII (500mg, 1.38mmol) is dissolved in 16ml anhydrous tetrahydro furan, addition potassium carbonate (191.4mg, 1.38mmol), DMSO-d6 (3.75ml), heavy water (2.5ml), 65 DEG C of oil baths are stirred to react 5 hours.LC-MS detection reaction terminates Afterwards, it removes tetrahydrofuran under reduced pressure, a small amount of water is added, is extracted with ethyl acetate, then organic phase is washed with water twice, saturated salt solution It washes once, evaporating solvent under reduced pressure obtains pure product 278mg, yield 55.6% through column chromatography for separation.1HNMR(400MHz, MeOD)δ0.93(-CH₃,s,3H).0.95(-CH₃,s,3H),1.29(-CH,s,1H),1.41-1.36(-CH₂, m, 2H), 2.78 (-CH,dd,J₁=8.9, J₂=6.3Hz, 1H), 7.49 (- Ph, d, J=1.4Hz, 2H), 7.61 (- Ph, t, J=1.3Hz, 1H), MS (ESI): observed:m/z 375 [M-H]^-。

((R) -1- (two deuterium acetylamino of 2- (2,5- benzamido) -2,2-) -3- methyl butyl) borocitric acid ester (NNU-459) preparation:

Citric acid (192.12mg, 0.39mmol) is dissolved in 2mL ethyl acetate, is warming up to 74 DEG C, it is complete to citric acid After fully dissolved, the compound VII (363.03mg, 0.36mmol) for being dissolved in 1mL ethyl acetate is added, slow cooling is to 60 DEG C, reaction 3h, then slow cooling to 25 DEG C overnight.TLC detection reaction, filtering, filter cake vacuum drying obtain pure products 90.0mg, yield 48.6%.¹H NMR(400MHz,DMSO)δ0.86(-CH₃, d, J=6.3Hz, 6H), 1.39-1.21 (- CH₂, m, 2H), 1.70 (- CH, d, J=26.1Hz, 1H), 2.81-2.52 (- CH₂,m,4H),2.88(-CH,s,1H),7.78-7.44(-Ph,m,3H), 9.12(-NH,s,1H),10.73(-NH,s,1H),12.15(-COOH,s,2H).MS(ESI): observed:m/z 519[ M]⁺。

Embodiment 2:((R) -1- (two deuterium acetylamino of 2- (2,5- benzamido) -2,2-) -3- methyl butyl) boric acid Diethanol amine ester (compound N NU-458)

According to method described in embodiment 1, the difference lies in that citric acid is replaced with diethanol amine, so that targeted be made Close object.¹H NMR(400MHz,DMSO)0.85-0.74(-CH₃,m,6H),1.33-1.16(-CH₂,m,2H),1.64-1.51(- CH,m,1H),2.98-2.63(-CH₂,m,4H),3.13(-CH,td,J₁=14.0Hz, J₂=7.1Hz, 1H), 3.67 (- CH₂, dd,J₁=11.4Hz, J₂=5.8Hz, 4H), 5.24 (- NH, t, J=22.6Hz, 1H), 6.58 (- NH, d, J=28.1Hz, 1H), 7.01 (- NH, d, J=8.4Hz, 1H), 7.65-7.43 (- Ph, m, 3H), 8.79 (d, J=36.3Hz, 1H) .MS (ESI): observed:m/z 432.3[M+H]⁺

Embodiment 3: rat pharmacokinetic evaluation

SD male rat 12,220 ± 20g of weight is randomly divided into four groups.

Wherein press 0.500mgkg for two groups^-1Tail vein injection gives 0.100mgmL to dosage respectively^-1NNU-459 and MLN-9708, and 10min, 20min before administration and after administration, 30min, 1h, 2h, 4h, 8h, 12h, for 24 hours and 36h, it is quiet by neck Arteries and veins blood sampling about 0.200mL, is placed in equipped with EDTA-K₂Test tube in, separated plasma after high speed centrifugation (7800 × g) 5min, in -15 DEG C~-35 DEG C of preservations.For comparing the pharmacokinetic difference of NNU-459 Yu MLN-9708 intravenous administration.

Other two groups are pressed 1.50mgkg^-1Dosage 0.150mgmL is given in stomach-filling respectively^-1NNU-459 and MLN- 9708, and 5min, 10min before administration and after administration, 20min, 30min, 1h, 2h, 4h, 8h, 12h, for 24 hours and 36h, it is quiet by neck Arteries and veins blood sampling about 0.200mL, is placed in equipped with EDTA-K₂Test tube in, separated plasma after high speed centrifugation (7800 × g) 15min, in- 15 DEG C~-35 DEG C preservations.For comparing the pharmacokinetic difference of NNU-459 Yu MLN-9708 oral administration.

The results show that whether intravenous administration or be administered orally NNU-459 half-life period will better than MLN-9708, Especially oral administration approach, the half-life period of NNU-459 are that (13.2 ± 7.97 hours) are apparently higher than partly declining for MLN-9708 Phase (5.80 ± 1.33 hours).

Find out from result above, the compounds of this invention has better pharmacokinetics in animal body, thus has more Good pharmacodynamics and therapeutic effect.

In addition, by deuterate, metabolic process of the compounds of this invention in organism is changed.This leads to first pass effect The reduction of (First-pass effect).In such a case, it is possible to change dosage and form durative action preparation, can also grow The form for imitating preparation improves applicability.

In addition, pharmacokinetics effect is also changed by deuterate, because deuterated compound forms another hydrate completely Film, so that the distribution in organism differs markedly from not deuterated compound.

The hepatomicrosome stability experiment of embodiment 4:NNU-455 and MLN-2238

Measurement in the hepatomicrosome of different plant species, the activity in vivo substance NNU-455 of NNU-459 and MLN-9708 with The stability of MLN-2238.

The preparation of hepatomicrosome and NADPH coenzyme:

I. people's hepatomicrosome working solution (2 ×): people's hepatomicrosome storing liquid (20mg/mL) of 0.3mL is taken to be added to 5.7mL phosphate buffer obtains the working solution of 1.0mg/mL.

Ii. rat liver microsomes working solution (2 ×): the rat liver microsomes storing liquid (20mg/mL) of 0.3mL is taken to be added To 5.7mL phosphate buffer, the working solution of 1.0mg/mL is obtained.

Iii. Mouse Liver Microsomes working solution (2 ×): take 0.3mL Mouse Liver Microsomes storing liquid (20mg/mL) plus Enter the working solution that 1.0mg/mL is obtained to 5.7mL phosphate buffer.

Iv. dog hepatomicrosome working solution (2 ×): the dog hepatomicrosome storing liquid (20mg/mL) of 0.3mL is taken to be added to 5.7mL phosphate buffer obtains the working solution of 1.0mg/mL.

V. monkey hepatomicrosome working solution (2 ×): the monkey hepatomicrosome storing liquid (20mg/mL) of 0.3mL is taken to be added to 5.7mL phosphate buffer obtains the working solution of 1.0mg/mL.

The working solution (2.5 ×) of vi.NADPH coenzyme: take the NADP (100mM) of 600 μ L, the G6P (200mM) of 600 μ L, The magnesium chloride solution (300mM) of the G6PDH (250U/mL) and 480 μ L of 240 μ L are added to the phosphate buffer of 18.08mL, obtain The working solution of NADPH coenzyme.

The preparation of working solution

I. the working solution (10 ×) of positive control: Verapamil is made into 20mM stock solution as positive control, with DMSO, 10 μM of working solutions are diluted to phosphate buffer again.

Ii. the working solution (10 ×) of test article: being first made into 10mM stock solution with DMSO, then with 50% methanol-water 100 μM are diluted to, then is diluted to the working solution of 10 μM of concentration with phosphate buffer.

The preparation of terminate liquid: methanol is used to configure 100ng/mL orinase as the terminate liquid of containing the internal standard.

Matrix needed for reacting is added according to the following table 1 sequence, test article each time point does 3 in parallel, and positive control is every A time point does 3 in parallel.

After the reaction was completed, by sample at 4 DEG C, 4000rpm is centrifuged 20min.The supernatant of 100 μ L is taken to use plus 200 μ L methanol In mass spectrum quantitative analysis.Concentration Aa/Ai (compound and internal standard mass signal peak area of the compound in different incubation time points Ratio) indicate.The concentration of same compound each time point is obtained into surplus ratio divided by 0 time point concentration, calculation formula is such as Under:

Half-life period:

The remaining percentage of different time points is mapped with Exponential Regression Model, calculates half with following first order kinetics formula It declines the phase:

Clearance rate (CL):

Wherein, Vd is apparent partition coefficient

Experimental result is shown in Table 1, and in people's hepatomicrosome, the half-life period (108.3min) of NNU-455 will be considerably longer than MLN- 2238 half-life period (57.3min), while the clearance rate (12.8 μ L/min/mg) of Y455 is also considerably less than MLN-2238 (24.2 μ L/min/mg)。

The stability experiment result of the NNU-455 in different plant species hepatomicrosome of table 1

Above the experimental results showed that, the present invention in compound N NU-459 it is more difficult in vivo be metabolized removing, thus Possess better dynamic metabolism property and drug effect.

Embodiment 5: the Activity determination of compound inhibition tumor cell in vitro

The detection liquid that this patent utilizes is that single Solution Cell Proliferation detects box, comes from Promega company；Cell used is U266, RPMI8226.Experimental system is 110 μ L, wherein containing 90 μ L of cell suspension, detects 10 μ L of liquid, 10 μ of drug (inhibitor) L, final concentration of 4.54 × 10^-8M~1.77 × 10^-9M, the last one concentration are 0M, and actual disposition concentration is 5 × 10^-7M~ 1.95×10^-8M, the last one concentration are 0M.Specific experiment process is as follows:

1, medicine ordinance:

Precise drug is added DMSO and is dissolved to 10^-2M.With pipettor draw 1 μ L add to 199 μ L DMSO obtain 5 × 10^-5M, then from 5 × 10^-5Drawn in M acute drug 3.3 μ L add the RPMI1640 culture medium of 326.7 μ L serum-frees obtain 5 × 10^-7M, 1.5 times of gradient dilutions, obtains 3.3 × 10^-7M、2.2×10^-7M、1.48×10^-7M、9.87×10^-8M、6.58×10^{- 8}M、4.38×10^-8M、2.92×10^-8M、1.95×10^-8The drug of M concentration, the last one concentration 0M are not dosing.

2, cell suspension configures:

After cell counts respectively, dilution configuration U266 is 1 × 10⁴A/hole, RPMI8226 are 1 × 10⁴A/hole.

3, prepared by reaction system:

90 μ L of cell suspension is added in every hole in 96 hole fluorescence ELISA Plates, is incubated for for 24 hours；Then 10 μ L are added in every hole and wait for test sample Product are positive control drug using compound MLN-9708, are incubated for for 24 hours；After completion of the reaction, every hole is added 10 μ L and detects liquid, is incubated for 2-3h, 490nm fluorescence microplate reader (BMG LABTECH POLARstar OPTIMA Microplate Reader) detects extinction Degree.

4, data processing

The absorbance of products therefrom under various concentration drug effect after deducting background is calculated, it is soft with GraphPad Prism Part calculates drug to the IC of cytotoxicity₅₀Concentration.

5, result

As shown in table 2, the different multiple myeloma cell lines (U266, RPMI8226) of two kinds cultivated in vitro, and In human breast cancer cell line MAD-MB-231 cell.The IC of NNU-459, NNU-458₅₀It is similar to MLN-9708, show phase Activity that is same or preferably inhibiting tumour.

The activity of table 2 compound N NU-458, NNU-459 and MLN-9708 to different tumor cell lines

Embodiment 6: pharmaceutical composition

Drug in the present invention can be taken but be not limited to following pharmaceutical composition:

Pharmaceutical composition 1

Ingredient	Per unit dose dosage (mg)	Dosage (%w/w or w/w)
			The compound	4	1.3
Silicified microcrystalline cellulose	292.4	97.5
			Talcum powder	0.6	0.2
Magnesium stearate	3	1
			Content weight	300	100

Pharmaceutical composition 2

Ingredient	Per unit dose dosage (mg)	Dosage (%w/w or w/w)
			The compound	4	1.3
Microcrystalline cellulose	231.8	77.3
			Pregelatinized starch	60	20
Silica	1.2	0.4
			Magnesium stearate	3	1
Content weight	300	100

Pharmaceutical composition 3

Ingredient	Per unit dose dosage (mg)	Dosage (%w/w or w/w)
			The medicament	4	1.3
Microcrystalline cellulose	246.8	82.3
			Mannitol	45	15
Talcum powder	1.2	0.4
			Magnesium stearate	3	1
Content weight	300	100

Pharmaceutical composition 4

Ingredient	Per unit dose dosage (mg)	Dosage (%w/w or w/w)
			The compound	3	1.5
Microcrystalline cellulose	150	75
			Cornstarch	45	22.5
Magnesium stearate	2	1
			Content weight	200	100

Pharmaceutical composition 5

Pharmaceutical composition 6

Ingredient	Per unit dose dosage (mg)	Dosage (%w/w or w/w)
			The compound	3	1.5
Mannitol	100	50
			Microcrystalline cellulose	95	47.5
Magnesium stearate	2	1
			Content weight	200	100

Pharmaceutical composition 7

Ingredient	Per unit dose dosage (mg)	Dosage (%w/w or w/w)
			The compound	2.3	1.15
Microcrystalline cellulose	195	97.5
			Talcum powder	0.7	0.35
Magnesium stearate	2	1
			Content weight	200	100

Pharmaceutical composition 8

Ingredient	Per unit dose dosage (mg)	Dosage (%w/w or w/w)
			The compound	2.3	1.15
Silicified microcrystalline cellulose	155	77.5
			Mannitol	40	20
Talcum powder	0.7	0.35
			Magnesium stearate	2	1
Content weight	200	100

According to a conventional method, after aforementioned pharmaceutical compositions being mixed in proportion, it is packed into opaque white gelatin glue In capsule.

In the present embodiment, " compound " includes compound N NU-455 and its citric acid ester (NNU-459) and diethyl Alkanolamine ester (NNU-458).

All references mentioned in the present invention is incorporated herein by reference, independent just as each document It is incorporated as with reference to such.In addition, it should also be understood that, after reading the above teachings of the present invention, those skilled in the art can To make various changes or modifications to the present invention, such equivalent forms equally fall within model defined by the application the appended claims It encloses.

Claims (8)

1. a kind of deuterated dipeptide boronic acid or boric acid ester compound, shown in structure such as following formula (a),

Wherein:

R₁And R₄It is halogen, R₂And R₃It is hydrogen, R₅And R₆It is deuterium；

P group is boronate, or forms borate or anhydride group by following segment and alpha-hydroxy carboxylic acid compounds or beta-hydroxycarboxylic acids Group, or by following segment and the compound with the not independent hydroxyl structure of two or more on same carbon atom Borate group is formed, the 'alpha '-hydroxy acids or beta-hydroxy acid are selected from glycolic, malic acid, hexahydro mandelic acid, 2- hydroxy-isobutyric Acid, citric acid, mandelic acid, lactic acid, 3- hydroxy-butyric acid, beta-hydroxy isovaleric acid, 2- hydroxyl -3,3- acid dimethyl, 2- hydroxyl - 3 Methylbutanoic acid ,-hydroxy isocaproic acid, tartaric acid, salicylic acid or benzoic acid；It is described have two not on same carbon atom or The compound of more than two independent hydroxyl structures is selected from diethanol amine, tartaric acid, alpha-D-glucose or D-ribose；

Wherein B is boron atom, and the atom connecting with formula (a) is B.

2. a kind of deuterated dipeptide boronic acid or boric acid ester compound, which is characterized in that the compound name is ((R) -1- (bis- deuterium acetylamino of 2- (2,5- benzamido) -2,2-) -3- methyl butyl) boric acid (compound N NU-455), chemical structure Formula is as follows:

3. a kind of deuterated dipeptide boronic acid or boric acid ester compound, which is characterized in that the compound name is ((R) -1- (two deuterium acetylamino of 2- (2,5- benzamido) -2,2-) -3- methyl butyl) boric acid diethanol amine ester (compound N NU- 458), chemical structural formula is as follows:

4. a kind of deuterated dipeptide boronic acid or boric acid ester compound, which is characterized in that the compound name is ((R) -1- (bis- deuterium acetylamino of 2- (2,5- benzamido) -2,2-) -3- methyl butyl) borocitric acid ester (compound N NU-459), Chemical structural formula is as follows:

5. the preparation method of claim 1-4 described in any item deuterated dipeptide boronic acids or boric acid ester compound, feature It is, including reaction process as shown below:

Wherein, R₁、R₂、R₃、R₄、R₅And R₆With P group with claim 1.

6. claim 1-4 described in any item deuterated dipeptide boronic acids or boric acid ester compound are preparing protease inhibition body Application in drug.

7. claim 1-4 described in any item deuterated dipeptide boronic acids or boric acid ester compound treat and prevent egg in preparation Application in white enzyme body related disease drug.

8. claim 1-4 described in any item deuterated dipeptide boronic acids or boric acid ester compound treat or prevent cancer in preparation Application in disease, cardiovascular disease, inflammation, immunity disease, nephrosis, angiogenesis or prostatic disorders drug.