CN106916177A - A kind of deuterated dipeptide boronic acid or its ester type compound and its synthetic method and purposes - Google Patents

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

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Publication number
CN106916177A
CN106916177A CN201710178276.2A CN201710178276A CN106916177A CN 106916177 A CN106916177 A CN 106916177A CN 201710178276 A CN201710178276 A CN 201710178276A CN 106916177 A CN106916177 A CN 106916177A
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deuterated
compound
acid
boronic acid
boric acid
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CN106916177B (en
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朱永强
雷萌
白恩赫
冯华云
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Nanjing mausoleum Medical Technology Co., Ltd.
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Nanjing Normal University
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Priority to PCT/CN2018/087761 priority patent/WO2018171816A1/en
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F5/00—Compounds containing elements of Groups 3 or 13 of the Periodic System
    • C07F5/02—Boron compounds
    • C07F5/025—Boronic and borinic acid compounds
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B59/00—Introduction of isotopes of elements into organic compounds ; Labelled organic compounds per se
    • C07B59/001—Acyclic or carbocyclic compounds
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B59/00—Introduction of isotopes of elements into organic compounds ; Labelled organic compounds per se
    • C07B59/002—Heterocyclic compounds
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B59/00—Introduction of isotopes of elements into organic compounds ; Labelled organic compounds per se
    • C07B59/005—Sugars; Derivatives thereof; Nucleosides; Nucleotides; Nucleic acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H23/00—Compounds containing boron, silicon, or a metal, e.g. chelates, vitamin B12
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B2200/00—Indexing scheme relating to specific properties of organic compounds
    • C07B2200/05—Isotopically modified compounds, e.g. labelled

Abstract

The invention discloses a kind of deuterated dipeptide boronic acid or boric acid ester compound or its crystal formation or pharmaceutically acceptable hydrate or solvate, shown in its 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 treatment or pre- anti-cancer, angiocardiopathy, inflammation, immunity disease, ephrosis, blood vessel generation 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 field, and in particular to a kind of deuterated dipeptide boronic acid or its ester type compound and its conjunction Into method and purposes.
Background technology
Ubiquitin-Proteasome Pathway (Ubiquitin-Proteasome Pathway, abbreviation UPP) is intracellular protein The main path of system degraded, and many physiologically important cellular processes, including signal transduction, immune response are participated in, do not roll over Folded albumino 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 relation.For this new target spot, first proteasome inhibitor bortezomib (PS- in 2003 341) FDA approval listings are obtained, for treating recurrent myeloma.The medicine in 2004 goes through to be listed in European Union again, for multiple Property myeloma.In September, 2005, the medicine is introduced by Xi'an Yang Sen, first in China's Guangzhou listing.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 reaches 30.69 hundred million dollars within 2014, as before global best-selling tumour medicine one of 20.Bortezomib is in China Market price is every 3.5 milligrams about 13000 yuan, about 40,000 yuan for the treatment of a cycle expense, and so expensive expense is for many trouble Person is very heavy financial burden.The U.S. FDA of in December, 2015 have approved first oral protein enzyme body inhibitor again Ixazomib is listed, for treating Huppert's disease.But current clinical data shows, it is many that this kind of medicine there is also comparing Side effect, such as fatigue it is weak, nauseous, diarrhoea and DPN.And, therefore, how to develop it is a kind of it is cheap, The low therapeutic protein enzyme body inhibitor oral drugs high of toxic and side effect are that we need key problems-solving at present.
The content of the invention
Goal of the invention：For above-mentioned technical problem, one class of the object of the invention offer is new to live with proteasome suppression The compound and its preparation method and use of property and more preferable pharmacodynamics performance.
Technical scheme：First aspect present invention disclose a kind of deuterated dipeptide boronic acid or boric acid ester compound or its Crystal formation or pharmaceutically acceptable hydrate or solvate, shown in its 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 groups are selected from following fragment：
Wherein, B is boron atom, Z1、Z2Independently selected from hydroxyl, C1-10Alkoxy or aryloxy group；
Or P groups are the heterocyclic groups containing N, S and/or O formed by following fragment and other compounds,
Wherein B is boron atom, and the atom being connected with formula (a) is B.
It is preferred that, the R1And R4It is halogen, R2、R3、R5、R6In have 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 groups are to form borate or anhydride group by following fragment and alpha-hydroxy carboxylic acid compounds or beta-hydroxycarboxylic acids,
Wherein B is boron atom, and the atom being connected with formula (a) is B.
It is preferred that, P groups are by following fragment and with two or more independent hydroxyls not on same carbon atom The compound of structure forms borate group,
Wherein B is boron atom, and the atom being connected with formula (a) is B.
In another preference, the deuterated deuterium isotopic content in deuterium the position of substitution is at least containing more than natural deuterium isotope Amount (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 more than 99%.
In another preference, formula (I) compound at least contains 1 D-atom, more preferably 2 D-atoms.
In another preference, R1、R4Selected from halogen；More preferably chlorine；
In another preference, R2、R3It is hydrogen；
In another preference, R5、R6Separately it is selected from hydrogen, deuterium, deuterated methyl, deuterated ethyl, halogen.
Borinate ester moiety is five-membered ring in certain 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 " be multiple groups be respectively selected from some substitution bases, 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 R2Do not connected each other between group.
As used herein, term " alpha-hydroxy carboxylic acid compounds " refers to contain the alpha position being directly connected to relative to hydroxy-acid group The compound of the hydroxyl on carbon atom.As used herein, term " alpha-hydroxy carboxylic acid compounds " be not limited to only to have hydroxyl and One compound of hydroxy-acid group.
As used herein, term " beta-hydroxycarboxylic acids " refers to contain the carbon being directly connected to relative to β of hydroxy-acid group The compound of the hydroxyl on atom.As used herein, term " beta-hydroxycarboxylic acids " is not limited to only have a hydroxyl and one The compound of individual hydroxy-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.Some of beta-hydroxycarboxylic acids other Non-limiting examples include glucoheptonic acid, maltobionic acid, lactobionic acid and galactosaccharic acid.Some of beta-hydroxycarboxylic acids other non-limits Property example processed includes enemy's oxalic acid, 1- hydroxy-2-naphthoic acids and 3- hydroxy-2-naphthoic acids.
In some embodiments, 'alpha '-hydroxy acids or beta-hydroxy acid are selected from glycolic, malic acid, hexahydro mandelic acid, 2- hydroxyls Base isobutyric acid, citric acid, mandelic acid, lactic acid, 3- hydroxy-butyric acids, beta-hydroxy isovaleric acid, 2- hydroxyl -3,3- acid dimethyls, 2- Hydroxy-3-methyl butyric acid ,-hydroxy isocaproic acid, tartaric acid, salicylic acid and benzoic acid.
In some embodiments, with the change of two or more independent hydroxyl structures not on same carbon atom Compound 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 structures：
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.Do not limited by any chemical bonding theory, in such embodiments, formula (a) can also be by Formula (3) or (4) represent：
Do not limited by any chemical bond and theory in the presence of other also, 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 lived Property.
In another preference, said compound is the preferred compound being selected from the group；
((R) -1- (the deuterium acetylaminos of 2- (2,5- benzamidos) -2,2- two) -3- methyl butyls) boric acid；
((R) -1- (the deuterium acetylaminos of 2- (2,5- benzamidos) -2,2- two) -3- methyl butyls) boric acid-(+)-australene Alkane diol ester
((R) -1- (the deuterium acetylaminos of 2- (2,5- benzamidos) -2,2- two) -3- methyl butyls) borocitric acid ester
((R) -1- (the deuterated dimethyl-acetylaminos of 2- (2,5- benzamidos) -2,2-) -3- methyl butyls) boric acid Chinese holly Rafter acid esters
((R) -1- (the deuterium acetylaminos of 2- (2,5- benzamidos) -2,2- two) -3- methyl butyls) borate tartar acid esters
Tartaric acid ((R) -1- (the deuterium acetylaminos of 2- (2,5- benzamidos) -2,2- two) -3- methyl butyls) borate
((R) -1- (the deuterium acetylaminos of 2- (2,5- benzamidos) -2,2- two) -3- methyl butyls) boric acid alpha-hydroxypentyl Two acid esters
((R) -1- ((S) 2- (2,5- benzamidos) -2- deuteriums-acetylamino) -3- methyl butyls) boric acid malate
((R) -1- (the deuterium acetylaminos of 2- (2,5- benzamidos) -2,2- two) -3- methyl butyls) boric acid diethanol amine Ester
((R) -1- ((R) 2- (2,5- benzamidos) two deuteriums of -2,2--acetylamino) -3- methyl butyls) boric acid grape Sugar ester
((R) -1- ((R) 2- (2,5- benzamidos) two deuteriums of -2,2--acetylamino) -3- methyl butyls) Portugal of boric acid two Grape sugar ester
((R) -1- ((R) 2- (2,5- benzamidos) two deuteriums of -2--acetylamino) -3- methyl butyls) boric acid ribose ester
Second aspect present invention provide described deuterated dipeptide boronic acid or boric acid ester compound or its crystal formation or The preparation method of pharmaceutically acceptable hydrate or solvate, including course of reaction as shown below：
Wherein, P groups are the heterocyclic groups containing N, S and/or O formed by following fragment and other compounds,
Wherein B is boron atom, and the atom being connected with formula (a) is B.
A kind of particularly preferred compound of the invention is ((R) -1- (deuterium acetyl of 2- (2,5- benzamidos) -2,2- two Amino) -3- methyl butyls) boric acid diethanol amine ester (compound N NU-458).
Another particularly preferred compound of the invention is ((R) -1- (deuterium second of 2- (2,5- benzamidos) -2,2- two Acylamino-) -3- methyl butyls) borocitric acid ester
The invention provides their preparation method.
Methods described includes：
In the ethyl acetate of boiling, compound (VIII) is NNU-455, with diethanol amine (DEA) reaction, is dropped by slow Warm crystallization obtains end-product NNU-458；
In another preference, NNU-455 and citron acid reaction can be allowed by similar approach, generate NNU-459.
In another preference, compound (VIII) be prepared by the following method：
In atent solvent, heavy water, DMSO-d6And in the presence of alkali, compound (VII) reaction forms the compound (Ⅷ)。
In another preference, compound (VII) be prepared by the following method：
Compound (VI) under the action of an acid with isobutaneboronic acid reacting generating compound (VII)；
In another preference, compound (VI) be prepared by the following method：
In atent solvent, compound (IV) and compound (V) in the presence of condensing agent and alkali, dehydrating condensation shape Into the compound (VI).
In another preference, compound (IV) obtain 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 preference, compound (III) obtain by the following method：
In atent solvent, 2,5- dichlorobenzoic acids (I) and compound (II) are changed in the presence of condensing agent and alkali Compound (III)
In another preference, described alkali is selected from potassium carbonate, cesium carbonate, sodium hydride, hydrofining, potassium hydroxide, hydrogen-oxygen Change sodium or combination.
Peptide condensing agent common in above-mentioned reaction is N, and N- dicyclohexyl-carbodiimides (are abbreviated as DCC), 1- (3- diformazans Aminopropyl) -3- ethyl-carbodiimide hydrochlorides (being abbreviated as EDC.HCl), 1- hydroxy benzo triazoles (being abbreviated as HOBt) or Isobutyl chlorocarbonate.
In another preference, each reaction temperature is carried out at -20~100 DEG C.
In the third aspect of the present invention, there is provided a kind of method for preparing pharmaceutical composition, including step：Pharmaceutically will may be used The carrier of receiving and the compound described in first aspect present invention, or its pharmaceutically acceptable crystal formation, hydrate or solvent Compound is mixed, so as to form pharmaceutical composition.
In the fourth aspect of the present invention, there is provided a kind of pharmaceutical composition, it contains pharmaceutically acceptable carrier and sheet Compound described in invention first aspect, or its crystal formation, pharmaceutically acceptable hydrate or solvate.
In another preference, described pharmaceutical composition is injection, wafer, tablet, pill, powder or granule.
In another preference, described pharmaceutical composition also contains other medicine, described other treatment Medicine is cancer, angiocardiopathy, inflammation, immunity disease, ephrosis, blood vessel generation, the medicine of prostatic disorders.
More preferably, described medicine 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), support pool examination (etoposide), everolimus (everolimus), fulvestrant (fulvestrant), Gefitinib (gefitinib), 2, 2- difluoro deoxidations 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), the granulocyte colony stimulating factor of PEGylation (pegfilgrastin), the alpha-interferon (peglated alfa-interferon) of PEGylation, the U.S. song (pemetrexed) of training, 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 present invention, there is provided the compound described in first aspect present invention, or its crystal formation, pharmaceutically The purposes of acceptable salt, hydrate or solvate, they be used to prepare the pharmaceutical composition of protease inhibition body.
In another preference, described compound can be used for treating and preventing the disease related to proteasome target spot Disease.
In another preference, described pharmaceutical composition is used to treat and prevent following disease:Cancer, cardiovascular disease Disease, inflammation, immunity disease, ephrosis, blood vessel generation or prostatic disorders.
In another preference, described cancer includes (but being not limited to):Huppert's disease, non-small cell lung cancer, The cancer of the uterus, 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, there is provided a kind for the treatment of method, it includes step：Applied to the object for the treatment of is needed Compound described in first aspect present invention, or its crystal formation, pharmaceutically acceptable hydrate or solvate, or apply this Pharmaceutical composition described in the invention third aspect, so that protease inhibition body.It is preferred that described disease includes：Cancer, the heart Vascular diseases, inflammation, immunity disease, ephrosis, blood vessel generation or prostatic disorders.
It should be understood that in the scope of the invention, above-mentioned technical characteristic and each technical characteristic for hereinafter specifically describing can To be mutually combined, so as to constitute new or preferred technical scheme.As space is limited, no longer tire out one by one herein and state.
Specific embodiment
It has been investigated that, deuterated dipeptide boronic acid of the invention and borate and not deuterated compound phase ratio, with more excellent Different pharmacokinetics and/or pharmacodynamics performance, therefore the compound of protease inhibition body is more suitable for, and then be more suitable for Prepare the medicine for the treatment of cancer and relevant disease.The present invention is completed on this basis.
With ((R) -1- (the deuterium acetylaminos of 2- (2,5- benzamidos) -2,2- two) -3- methyl butyls) borocitric acid As a example by ester (compound N NU-459).
Pharmacokinetics in Rat experimental result shows, half-life period Ts of the NNU-459 than MLN97082/1Extension, under curve Area AUC is dramatically increased, and the apparent clearance rates of NNU-459 are also substantially reduced than MLN9708.Simultaneously in the hepatomicrosome stability of people In experiment, the half-life period (T of the activity in vivo composition (NNU-455) of compound N NU-4591/2=108.3min) also than Half-life period (the T of the active component (MLN2238) of MLN97081/2=57.3min) significantly increase.
In addition in testing in vitro, to tumor cell line U266, RPMI-8266 and breast cancer cell line MAD-MB-231 The IC of suppression50(nM) on, NNU-459 also shows more preferable 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.Deuterated can be one Substitution, two substitutions, polysubstituted or full substitution.Term " one or more are deuterated " is interchangeable with " one or many is deuterated " to be made With.
It is deuterated to replace unknown deuterium isotopic content to be at least containing more than natural deuterium isotope in deuterium in another preference Amount (0.015%), is preferably greater than 30%, more preferably more than 50%, even more preferably greater than 75%, even more preferably greater than 95%, more preferably Ground is more than 99%.
In another preference, formula compound at least contains 1 D-atom, more preferably 2 D-atoms.
Active component
Term " the compounds of this invention " as used herein refers to compound shown in formula (a).The term also includes and formula (a) chemical combination The various crystalline forms of thing, pharmaceutically acceptable hydrate or solvate.
Preparation method
The preparation method of formula (a) structural compounds, but these specific methods is described more particularly below not to this Invention constitutes any limitation.The compounds of this invention can also optionally will be describing or known in the art various in this manual Synthetic method is combined and is easily obtained, and such combination can readily be entered by those skilled in the art in the invention OK.
The not deuterated dipeptide boronic acid and its preparation method of esters that the present invention is used are known.The deuterated dipeptides of correspondence Boric acid and its esters can be raw material with corresponding deuterated boronic acid compounds.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-benzoyls base) glycine methyl ester (compound III)；Then compound III is in the presence of alkali Generation saponification obtains (2,5- dichloro-benzoyls base) glycine (compounds Ⅳ)；Compounds Ⅳ with (aR, 3aS, 4S, 6S, 7aR)-hexahydro -3a, 8,8- trimethyl-alpha- (2- methyl-propyls) -4,6- first bridge -1,3,2- benzo dioxy borine -2- methylamines In the presence of peptide condensing agent and alkali, reaction generates ((R) -1- (2- (2,5- benzene first to 2,2,2- trifluoroacetates (compound V) Amide groups) the deuterium acetylaminos of -2,2- two) -3- methyl butyls) boric acid-(+)-australene alkane diol ester (compound VI)；In methyl alcohol In, compound 5 and isobutaneboronic acid reaction generation ((R) -1- (2- (2,5- benzamido)-acetyl in the presence of 1N hydrochloric acid Amino) -3- methyl butyls) boric acid (compound VII)；Then in tetrahydrofuran, in potassium carbonate and deuterated DMSO and the work of heavy water Under, ((R) -1- (the deuterium acetylaminos of 2- (2,5- benzamido) -2,2- bis-) -3- methyl butyls) boric acid (compound is generated Ⅷ)；Compound VII obtains compound N NU-459 in the ethyl acetate of boiling with citron acid reaction.
Specific synthetic method is discussed in detail in embodiment 1.
Pharmaceutical composition and application process.
Because the compounds of this invention has an excellent inhibitory activity to proteasome, therefore the compounds of this invention and its each Crystal formation, pharmaceutically acceptable hydrate or solvate are planted, and it is the medicine of main active to contain the compounds of this invention Compositions can be used to treating, prevent and alleviating the disease by mediating proteasome.According to prior art, chemical combination of the present invention Thing can be used to treat following disease：Cancer, angiocardiopathy, obesity, diabetes etc..
Pharmaceutical composition of the invention comprising the compounds of this invention in the range of safe and effective amount or its be pharmacologically subjected to Salt and pharmacologically acceptable excipient or carrier.Wherein " safe and effective amount " is referred to：The amount of compound is enough to substantially Improve the state of an illness, and be unlikely to produce serious side effect.Generally, pharmaceutical composition contains 1-2000mg the compounds of this invention/agent, More preferably, 10-200mg the compounds of this invention/agent is contained.It is preferred that described is " one " for a capsule or tablet.
" pharmaceutically acceptable carrier " is referred to:One or more biocompatible solid or liquid filler or jello Matter, they are suitable for people and use and it is necessary to have enough purity and sufficiently low toxicity." compatibility " is referred to herein as combined In thing each component energy and compound of the invention and they between mutually admix, and significantly reduce the drug effect of compound.
Term " carrier ", " excipient " or " carrier " is 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, such as by producing any undesirable biological effect or any other component phase with pharmaceutical composition Interaction makes compound decreasing effect or failure, or generation harmful substance, otherwise its purposes be considered as in the scope of the present invention.
Pharmaceutically acceptable carrier includes but is not limited to ion-exchanger, aluminum 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, cataloid, magnesium trisilicate, polyvinylpyrrolidone, polyacrylate, Polyethylene-polyoxypropylene block polymer, lanolin, sugar such as lactose, glucose, sucrose and mannitol, starch such as cornstarch And farina, 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 propane diols 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 ethanol, isopropanol alcohol, hexadecanol and glycerine, cyclodextrin such as hydroxypropyl Beta-schardinger dextrin and sulfobutyl ether beta-schardinger dextrin, lubricant such as lauryl sodium sulfate and magnesium stearate, petroleum hydrocarbon such as mineral oil and Vaseline.Colouring agent, releasing agent, coating agent, sweetener, flavor enhancement 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 well known in the art, such as conventional granulation, mixing, molten Solution, encapsulating, lyophilized or emulsification method etc..Composition can be produced in a variety of manners, including particle, be precipitated or particle, powder, bag Include freeze-drying, Rotary drying or spray-dried powders, amorphous powder, tablet, capsule, syrup, suppository, injection, emulsion, Elixir, supensoid agent or solution.
The method of application of the compounds of this invention or pharmaceutical composition is not particularly limited, according to preferred embodiment, will Composition of the invention is formulated for mammal, preferably to the medicament administration of the mankind.These drug regimens of the invention Thing can be by oral, alimentary canal external administration, by suction spraying, part, rectum, nose, cheek, vagina or via implanted reservoir Using.Term " alimentary canal external administration " used herein includes subcutaneous, intravenous, intramuscular, in joint, intrasynovial, and in breastbone, sheath It is interior, with intracranial injection or infusion techniques in liver.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 be applied locally rather than systemic fashion, such as in tumour Apply (such as by injection) at position.
Liquid formulation for oral administration includes but is not limited to pharmaceutically acceptable emulsion, and microemulsion, solution is mixed Suspension, syrup and elixir.In addition to reactive compound, liquid dosage form can contain inert diluents commonly used in the art Agent, such as water or other solvents, solubilizer and emulsifying agent, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, Ergol, propane diols, 1,3-BDO, cyclodextrin, dimethylformamide, oil (particularly cottonseed oil, peanut oil, corn Oil, embryo oil, olive oil, castor oil and sesame oil), glycerine, tetrahydrofurfuryl alcohol, polyethylene glycol and fatty acid ester, anhydro sorbitol And its mixture.Besides inert diluents, Orally administered composition may also include adjuvant, such as wetting agent, emulsifying agent and suspending agent, sweet Taste agent, spin agent and aromatic.
Solid dosage forms for being administered orally includes capsule, tablet, pill, powder and granule.In these solid formulations In type, reactive compound mixes with least one conventional inert excipients (or carrier), such as sodium citrate or Dicalcium Phosphate, or with Following compositions mix:(a) filler or bulking agent, for example, starch, lactose, sucrose, glucose, mannitol and silicic acid；B () bonds Agent, for example, hydroxymethyl cellulose, alginates, gelatin, PVP, sucrose and Arabic gum；(c) NMF, example Such as, glycerine；(d) disintegrant, for example, agar, calcium carbonate, farina or tapioca, alginic acid, some 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 Alcohol and glycerin monostearate；(h) adsorbent, for example, kaolin；(i) lubricant, for example, talcum, calcium stearate, stearic acid Magnesium, solid polyethylene glycol, lauryl sodium sulfate, or its mixture.In capsule, tablet and pill, formulation also can be comprising slow Electuary.
Solid dosage forms such as tablet, sugar-pill, capsule, pill and granule can using be coated and shell material prepare, such as casing and Other materials well known in the art.They can include reactive compound or compound in opacifying agent, also, this composition Release can discharge in certain part in a delayed fashion in alimentary canal.The example of adoptable embedding component is polymeric material And Wax.If necessary, reactive compound also can form microencapsulation form with one or more in above-mentioned excipient.
In addition to these inert diluents, composition can also include auxiliary agent, such as wetting agent, emulsifying agent and suspending agent, sweet taste Agent, flavouring and spices.
Injectable formulation, such as sterile injectable be 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 emulsion, such as the solution in 1,3-BDO.What can be used is acceptable Carrier and solvent be water, compound sodium chloride injection, and isotonic sodium chlorrde solution.Additionally, aseptic expressed oi is generally used Make solvent or suspension media, it is possible to use any gentle expressed oi, including the monoglyceride or diglyceride for synthesizing.This Outward, aliphatic acid such as oleic acid is used to prepare injection.Injectable formulation can for example, by by bacteria retaining filter filter or Sterilized by mixing the bactericidal agent of aseptic solid composite form, the aseptic solid composite can use preceding dissolving or divide Dissipate in sterilized 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.
Formulation for part or applied dermally 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 of needs or the buffer solution that may be needed mix.Eye-drops preparations, auristilla and eye drops are recognized as of the invention In the range of.Additionally, the present invention considers to use transdermal patch, it has the additional excellent of the controlled delivery to body offer compound Point.Such formulation can be prepared by the way that compound is dissolved or dispersed in appropriate medium.Absorption enhancer also can use The flux of skin is passed through in increase compound.Can be by providing rate controlling membranes or being dispersed in polymer matrix by by compound Carry out speed control in matter or gel.
In some embodiments, the invention provides compound and the drug regimen of other excipient as herein described Thing.In some of the other embodiments, the invention provides the compound and the medicine of other excipient as herein described of NNU-455 Compositions.In other embodiments, the invention provides citric acid ester (NNU-459) and diethanol comprising NNU455 The pharmaceutical composition of amine ester (NNU-458) and other excipient as herein described.
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, there is provided the solid oral dosage form of the stabilization of reactive compound.
The compounds of this invention can be administered alone, or with other pharmaceutically acceptable compound administering drug combinations.
It is the mammal that the compounds of this invention of safe and effective amount is applicable to treatment during using pharmaceutical composition (such as people), wherein using when dosage be effective dosage for pharmaceutically thinking, for the people of 60kg body weight, day is to medicament Amount is usually 1~2000mg, preferably 20~500mg.Certainly, specific dosage is also contemplated that method of administration, patient health situation etc. Factor, within the scope of these are all skilled practitioners technical ability.
Compound of the invention and the compound phase ratio for not carrying deuterium well known in the prior art, with series of advantages：
(1) the compounds of this invention has excellent inhibition to proteasome.
(2) by the metabolism in deuterate this technology change organism, extend the metabolic half life of medicine, this causes head Cross the reduction of effect (First-pass effect).In such a case, it is possible to change dosage and form durative action preparation, its 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) with the hydrogen atom in deuterium substituted compound, due to its isotope effect, it is possible to increase compound is in animal body Drug concentration, to improve curative effect of medication.
With reference to specific embodiment, the present invention is expanded on further.It should be understood that these embodiments are merely to illustrate the present invention Rather than limitation the scope of the present invention.The experimental technique of unreceipted actual conditions in the following example, generally according to conventional strip Part, or according to the condition proposed by manufacturer.Unless otherwise indicated, otherwise parts and percentages are weight portion and weight percent Than.
Embodiment 1：(S) the deuterium acetamides of-N- (2,5- dichloro-benzoyls base) -2,2- two-D-Leu borocitric acid ester (compound N NU-459).Synthetic route：
(S) preparation of-N- (2,5- dichloro-benzoyls base) glycine (IV)：
2,5- dichlorobenzoic acids (7.6g, 40mmol) and HOBt (8.1g, 40mmol) are dissolved in CH2Cl2In (200mL), React 10min at -10 DEG C, add EDC.HCl (11.5g, 60mmol) reaction 30min, add compound ii (5g, 40mmol), DIPEA (18.1g, 140mmol) is added after reaction 10min, reaction is warmed to room temperature overnight after reaction 20min.TLC is examined Survey reaction, respectively with 10% hydrochloric acid solution (200mL), 5% NaHCO3Solution
(200mL) and saturated aqueous common salt (2 × 200mL) are washed, CH2Cl2Layer anhydrous Na2SO4Dry, filtering, decompression is steamed Except solvent, oily compound 9.32g is obtained, yield 88.9%.
Take step gained compound III (129mg, 0.31mmol) 2.5mLMeOH to dissolve, add LiOHH2O (39mg, 0.92mmol) and H2O (0.8mL), TLC detect that reaction is finished after 2h.Organic phase is spin-dried for, with ether (2 × 1mL) aqueous phase extracted, It is 2~3 that water is mutually added dropwise hydrochloric acid to pH value, produces a large amount of white solids, and ethyl acetate extraction removes 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-benzoyls 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) ,- React 10min at 10 DEG C, add EDC.HCl (321mg, 1.67mmol) reaction 30min, add (aR, 3aS, 4S, 6S, 7aR)- Hexahydro -3a, 8,8- trimethyl-alpha- (2- methyl-propyls) -4,6- first bridge -1,3,2- benzo dioxy borine -2- methylamines 2,2, 2- trifluoroacetates (317mg, 0.84mmol), add DIPEA (433mg, 3.35mmol), after reaction 20min after reaction 10min It is warmed to room temperature reaction overnight.TLC detection reaction, respectively with 10% hydrochloric acid solution (20mL), 5% NaHCO3Solution (20mL) With saturated aqueous common salt (2 × 20mL) washing, CH2Cl2Layer anhydrous Na2SO4Dry, filtering removes solvent under reduced pressure, through column chromatography 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- benzamidos)-acetylamino) -3- methyl butyls) 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 reactions, n-hexane Mutually extracted 2 times with MeOH (2 × 3mL), n-hexane (3mL) washing methyl alcohol phase 1 time removes methyl alcohol, CH under reduced pressure2Cl2(2 × 2mL) extracts Water intaking phase 2 times, organic phase is washed with saturated aqueous common salt (3 × 5mL) to water mutually in neutrality.Solvent is removed 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- (the deuterium acetylaminos of 2- (2,5- benzamidos) -2,2- two) -3- methyl butyls) boric acid (VIII) It is standby：
Compound VII (500mg, 1.38mmol) is dissolved in 16ml anhydrous tetrahydro furans, addition potassium carbonate (191.4mg, 1.38mmol), DMSO-d6 (3.75ml), heavy water (2.5ml), 65 DEG C of oil bath stirring reactions 5 hours.LC-MS detection reactions terminate Afterwards, remove tetrahydrofuran under reduced pressure, add a small amount of water, be extracted with ethyl acetate, then organic phase is washed with water twice, saturated aqueous common salt Wash once, remove solvent under reduced pressure, pure product 278mg, yield 55.6% are obtained 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- (the deuterium acetylaminos of 2- (2,5- benzamidos) -2,2- two) -3- methyl butyls) borocitric acid ester (NNU-459) preparation：
Citric acid (192.12mg, 0.39mmol) is dissolved in 2mL ethyl acetate, 74 DEG C are warming up to, treats that citric acid is complete After CL, addition is dissolved in the compound VII (363.03mg, 0.36mmol) of 1mL ethyl acetate, and slow cooling reacts to 60 DEG C 3h, then slow cooling to 25 DEG C overnight.TLC detection reactions, filtering, filter cake vacuum drying obtains 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- (the deuterium acetylaminos of 2- (2,5- benzamidos) -2,2- two) -3- methyl butyls) boric acid Diethanol amine ester (compound N NU-458)
According to method described in embodiment 1, difference is to replace citric acid with diethanol amine, so as to targeted is obtained Compound.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), (d, J=36.3Hz, the 1H) .MS (ESI) of 7.01 (- NH, d, J=8.4Hz, 1H), 7.65-7.43 (- Ph, m, 3H), 8.79: observed:m/z 432.3[M+H]+
Embodiment 3：Rat pharmacokinetic is evaluated
SD male rats 12,220 ± 20g of body weight is randomly divided into four groups.
Wherein two groups are pressed 0.500mgkg-1Tail vein injection gives 0.100mgmL to dosage respectively-1NNU-459 and MLN-9708, and in administration before and be administered after 10min, 20min, 30min, 1h, 2h, 4h, 8h, 12h, 24h 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.Pharmacokinetic difference for comparing NNU-459 and MLN-9708 intravenous administrations.
Other two groups are pressed 1.50mgkg-1Dosage gavage gives 0.150mgmL respectively-1NNU-459 and MLN- 9708, and in administration before and be administered after 5min, 10min, 20min, 30min, 1h, 2h, 4h, 8h, 12h, 24h 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.Pharmacokinetic difference for comparing NNU-459 and MLN-9708 oral administrations.
Result shows, whether the half-life period of intravenous administration or oral administration NNU-459 will be better than MLN-9708, Especially oral administration approach, the half-life period of NNU-459 is (13.2 ± 7.97 hours) partly declining apparently higher than MLN-9708 Phase (5.80 ± 1.33 hours).
Find out from result above, the compounds of this invention has more preferable pharmacokinetics in animal body, thus with more Good pharmacodynamics and therapeutic effect.
In addition, by deuterate, metabolic process of the compounds of this invention in organism is changed.This causes first pass effect The reduction of (First-pass effect).In such a case, it is possible to change dosage and form durative action preparation, it can also grow The form for imitating preparation improves applicability.
In addition, pharmacokinetics is also changed by deuterate act on, because deuterated compound forms another hydrate completely Film, so that the distribution in organism differs markedly from not deuterated compound.
Embodiment 4：The hepatomicrosome stability experiment of NNU-455 and MLN-2238
Determine in the hepatomicrosome of different plant species, the activity in vivo material 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) for taking 0.3mL is added to 5.7mL phosphate buffers, obtain the working solution of 1.0mg/mL.
Ii. rat liver microsomes working solution (2 ×)：The rat liver microsomes storing liquid (20mg/mL) for taking 0.3mL is added To 5.7mL phosphate buffers, the working solution of 1.0mg/mL is obtained.
Iii. Mouse Liver Microsomes working solution (2 ×)：The Mouse Liver Microsomes storing liquid (20mg/mL) for taking 0.3mL adds Enter to 5.7mL phosphate buffers, obtain the working solution of 1.0mg/mL.
Iv. dog hepatomicrosome working solution (2 ×)：Dog hepatomicrosome storing liquid (20mg/mL) for taking 0.3mL is added to 5.7mL phosphate buffers, obtain the working solution of 1.0mg/mL.
V. monkey hepatomicrosome working solution (2 ×)：Monkey hepatomicrosome storing liquid (20mg/mL) for taking 0.3mL is added to 5.7mL phosphate buffers, obtain 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 is added to the phosphate buffer of 18.08mL, obtains The working solution of NADPH coenzyme.
The preparation of working solution
I. the working solution (10 ×) of positive control：Verapamil is made into 20mM storing solutions as positive control with DMSO, Again 10 μM of working solutions are diluted to phosphate buffer.
Ii. the working solution (10 ×) of test article：First be made into 10mM storing solutions with DMSO, then with 50% methanol-water 100 μM are diluted to, then 10 μM of working solutions of concentration are diluted to phosphate buffer.
The preparation of terminate liquid：Terminate liquid of the 100ng/mL orinases as containing the internal standard is configured with methyl alcohol.
According to table 1 below order add reaction needed for matrix, each time point of test article do 3 it is parallel, positive control is every Individual time point do 3 it is parallel.
After the completion of reaction, by sample in 4 DEG C, 4000rpm centrifugations 20min.The supernatant for taking 100 μ L is used plus 200 μ L methyl alcohol 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) represent.The concentration at same each time point of compound is obtained into surplus ratio divided by 0 time point concentration, computing formula is such as Under：
Half-life period：
The remaining percentage of different time points is mapped with Exponential Regression Model, half is calculated with following first order kinetics formula Decline 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 test result indicate that, the present invention in compound N NU-459 it is more difficult in vivo be metabolized removing so that Possess more preferable dynamic metabolism property and drug effect.
Embodiment 5：Compound suppresses the Activity determination of tumor cell in vitro
The detection liquid that this patent is utilized is single Solution Cell Proliferation detection box, from Promega companies；Cell used is U266, RPMI8226.Experimental system is 110 μ L, wherein containing the μ L of cell suspension 90, detecting the μ L of liquid 10, the μ of medicine (inhibitor) 10 L, its final concentration of 4.54 × 10-8M~1.77 × 10-9M, last concentration is 0M, and actual disposition concentration is 5 × 10-7M~ 1.95×10-8M, last concentration is 0M.Specific experiment process is as follows：
1st, medicine ordinance：
Precise medicine, adds DMSO to be dissolved to 10-2M.With pipettor draw 1 μ L add to 199 μ L DMSO obtain 5 × 10-5M, then from 5 × 10-5The RPMI1640 culture mediums that 3.3 μ L plus 326.7 μ L serum-frees are drawn in M acute drugs 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- 8M、4.38×10-8M、2.92×10-8M、1.95×10-8The medicine of M concentration, last concentration 0M is not dosing.
2nd, cell suspension configuration：
After cell is counted respectively, dilution configuration U266 is 1 × 104Individual/hole, RPMI8226 is 1 × 104Individual/hole.
3rd, prepared by reaction system：
The μ L of cell suspension 90 are added in 96 hole fluorescence ELISA Plates per hole, 24h is incubated；Then 10 μ L are added to treat test sample in every hole Product, are positive control drug using compound MLN-9708, are incubated 24h；After completion of the reaction, 10 μ L detection liquid is added per hole, is incubated 2-3h, 490nm fluorescence microplate readers (BMG LABTECH POLARstar OPTIMA Microplate Reader) detection extinction Degree.
4th, data processing
The absorbance of the lower products therefrom of various concentrations medicine effect after background is deducted in calculating, soft with GraphPad Prism Part, calculates IC of the medicine to cytotoxicity50Concentration.
5th, result
As shown in table 2, the two kinds of different multiple myeloma cell lines (U266, RPMI8226) cultivated in vitro, and In MCF-7's MAD-MB-231 cells.The IC of NNU-459, NNU-45850It is similar to MLN-9708, show phase Activity that is same or preferably suppressing tumour.
Activity of table 2 compound N NU-458, NNU-459 and the MLN-9708 to different tumor cell lines
Embodiment 6：Pharmaceutical composition
Medicine in the present invention can be taken but be not limited to following pharmaceutical composition：
Pharmaceutical composition 1
Composition Per unit dose consumption (mg) Consumption (%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
Composition Per unit dose consumption (mg) Consumption (%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
Composition Per unit dose consumption (mg) Consumption (%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
Composition Per unit dose consumption (mg) Consumption (%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
Composition Per unit dose consumption (mg) Consumption (%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
Composition Per unit dose consumption (mg) Consumption (%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
Composition Per unit dose consumption (mg) Consumption (%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 are mixed in proportion, opaque white gelatin glue is loaded 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).
The all documents referred in the present invention are all incorporated as reference in this application, independent just as each document It is incorporated as with reference to such.In addition, it is to be understood that after above-mentioned instruction content of the invention has been read, those skilled in the art can Made various changes or modifications with to the present invention, these equivalent form of values equally fall within the model that the application appended claims are limited Enclose.

Claims (9)

1. a kind of deuterated dipeptide boronic acid or boric acid ester compound or its crystal formation or pharmaceutically acceptable hydrate or molten Agent compound, shown in its 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-C4Alkane Base；And R1、R2、R3、R4、R5、R6In at least one be deuterated or deuterium；
P groups are selected from following fragment：
Wherein, B is boron atom, Z1、Z2Independently selected from hydroxyl, C1-10Alkoxy, aryloxy group；
Or P groups are the heterocyclic groups containing N, S and/or O formed by following fragment and other compounds,
Wherein B is boron atom, and the atom being connected with formula (a) is B.
2. deuterated dipeptide boronic acid according to claim 1 or boric acid ester compound or its crystal formation or can pharmaceutically connect The hydrate or solvate received, it is characterised in that the R1And R4It is halogen, R2、R3、R5、R6In have two hydrogen and two Deuterium.
3. deuterated dipeptide boronic acid according to claim 1 or boric acid ester compound or its crystal formation or can pharmaceutically connect The hydrate or solvate received, it is characterised in that the R1And R4It is chlorine, R2And R3It is hydrogen, R5And R6It is deuterium.
4. deuterated dipeptide boronic acid according to claim 1 or boric acid ester compound or its crystal formation or can pharmaceutically connect The hydrate or solvate received, it is characterised in that P groups are formed with alpha-hydroxy carboxylic acid compounds or beta-hydroxycarboxylic acids by following fragment Borate or anhydride group,
Wherein B is boron atom, and the atom being connected with formula (a) is B.
5. deuterated dipeptide boronic acid according to claim 1 or boric acid ester compound or its crystal formation or can pharmaceutically connect The hydrate or solvate received, it is characterised in that P groups are by following fragment and have two not on same carbon atom Or the compound of two or more independence hydroxyl structure forms borate group,
Wherein B is boron atom, and the atom being connected with formula (a) is B.
6. deuterated dipeptide boronic acid or boric acid ester compound or its crystal formation described in any one of claim 1-5 or pharmaceutically The preparation method of acceptable hydrate or solvate, it is characterised in that including course of reaction as shown below：
Wherein, P groups are the heterocyclic groups containing N, S and/or O formed by following fragment and other compounds,
Wherein B is boron atom, and the atom being connected with formula (a) is B.
7. deuterated dipeptide boronic acid or boric acid ester compound or its crystal formation described in any one of claim 1-5 or pharmaceutically The application of acceptable hydrate or solvate in protease inhibition body medicine is prepared.
8. deuterated dipeptide boronic acid or boric acid ester compound or its crystal formation described in any one of claim 1-5 or pharmaceutically Acceptable hydrate or solvate are preparing the application in treating and preventing proteasome relevant disease medicine.
9. deuterated dipeptide boronic acid or boric acid ester compound or its crystal formation described in any one of claim 1-5 or pharmaceutically Acceptable hydrate or solvate prepare treatment or pre- anti-cancer, angiocardiopathy, inflammation, immunity disease, ephrosis, Application in blood vessel generation or prostatic disorders medicine.
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