TW201620515A - Single enantiomer as histone deacetylase inhibitor and preparation thereof - Google Patents

Abstract

This invention relates to novel Histone deacetylases inhibitors having the following structures: Also disclosed is a method for treating mucositis or cancer with these inhibitors.

Description

Single mirror phase isomer as tissue protein deacetylase inhibitor and preparation method thereof

This invention relates to novel tissue protein deacetylase inhibitors, and methods of using the same to treat mucosal ulcers and cancer.

Because tissue protein deacetylase inhibitors have the potential to treat cancer and inflammation, they are currently receiving much attention.

Tissue protein deacetylase inhibitors inhibit cancer cell growth while promoting cancer cell differentiation and apoptosis (Lu et al., J. Med. Chem. 2005, 48, 5530-5535; Kulp et al., Clin .Cancer Res. 2006, 12, 5199-5206; and Ryan et al., J. Clin. Onclo. 2005, 23, 3912-3922.). At the same time, it has been reported that this anti-acetylase inhibitor inhibits the expression of pro-inflammatory cytokines, thereby slowing down the inflammatory response, such as mucosal ulcers caused by cancer chemotherapy or radiation therapy (see IMAdcock, British Journal of Pharmacology, 2007, 150). (7): 839-831; and YL Chung et al., Carcinogenesis 2009 30(8): 1387-1397.).

The inventive team has successfully developed tert-butyl 3-(5-(benzyloxy)pyridin-2-yl)-1-(4-(4-(hydroxylamino)-4-oxobutyl)anilino)-1 - Racemic mixture of oxypropyl-2-ylcarbamate ((±)Aza-PBHA) in oral mucosal ulcer hamster test induced by cancer chemotherapeutic drug 5-FU, wound healing treatment effect of 90%; in cancer In the mouse test of gastrointestinal mucosal ulcer induced by cisplatin, the wound healing effect is equivalent to palifermin [HP Wang, et al., US Patent No. 8,242,282 B2]. The content of this U.S. patent is incorporated herein by reference.

A primary object of the invention resides in the third butyl 3-(5-(benzyloxy)pyridin-2-yl)-1-(4-(4-(hydroxyamino)-4-oxobutyl)anilino) A racemic mixture of (-1-)Aza-PBHA of 1-oxopropyl-2-ylcarbamate isolated (R)-(-)-t-butyl 3-(5-() Benzyloxy)pyridin-2-yl)-1-(4-(4-(hydroxyamino)-4-oxobutyl)anilino)-1-oxopropyl-2-ylcarbamate (R-( -)-Aza-PBHA) and (S)-(+)-t-butyl 3-(5-(benzyloxy)pyridin-2-yl)-1-(4-(4-(hydroxylamino)-4) Novel single mirror isomer of -oxybutyl)anilino)-1-oxopropyl-2-ylcarbamate (S-(+)-Aza-PBHA):

Another object of the present invention is to provide a (±) Aza-PBHA A method in which the racemic mixture is separated into R-(-)-Aza-PBHA and S-(+)-Aza-PBHA.

The present invention is based on the discovery that R-(-)-Aza-PBHA and S-(+)-Aza-PBHA are effective tissue protein deacetylase inhibitors, and have an activity of inhibiting mucosal ulceration. Accordingly, the present invention relates to R-(-)-Aza-PBHA and S-(+)-Aza-PBHA compounds themselves, and R-(-)-Aza-PBHA and S-(+)-Aza-PBHA in Tissue protein deacetylase-related disease applications.

The compounds described in the present invention comprise the compound itself, its salts, solubilities, and prodrugs. For example, the salt may be a compound formed of an anion and a positively charged group such as an amine group. Suitable anions include chloride, bromide, iodide, sulfate, hydrogen sulfate, amine sulfonate, nitrate, phosphate, citrate, mesylate, trifluoroacetate, Glutamate, glucuronide, glutarate, malate, maleate, succinate, fumarate, tartrate, tosylate, salicylate , lactate ion, naphthalene sulfonate ion and acetate ion. Likewise, salts may be formed from cations and negatively charged groups (eg, carboxylate ions) on sulfhydryl or dihydroanthracene compounds. Suitable cations include sodium ions, potassium ions, magnesium ions, calcium ions, ammonium (such as tetramethylammonium) ions. The above compounds simultaneously contain salts formed by quaternary nitrogen. Examples of prodrugs include esters and other pharmaceutically acceptable derivatives which, when applied to the subject, provide the above-described active compounds.

In still another aspect, the present invention relates to an effective agent for any of the above R-(-)-Aza-PBHA and S-(+)-Aza-PBHA A method of inhibiting tissue protein deacetylase (HDAC) activity by contacting cells.

In still another aspect, the present invention relates to an effective amount of any of the above R-(-)-Aza-PBHA and S-(+)-Aza-PBHA required for the treatment of a body for the treatment of mucosal ulcers And methods of treating cancer.

The present invention also encompasses a pharmaceutical composition comprising one or more of the above R-(-)-Aza-PBHA and S-(+)-Aza-PBHA for use in the treatment of mucosal ulcers or cancer; The therapeutic use of cancer, and the use of a medicament for treating mucosal ulcers or cancer.

Details of one or more specific embodiments of the invention are detailed below. Other features, objects, and advantages of the invention will be apparent from the description and appended claims.

Figure 1 is a chromatogram of the intermediate optical isomer R-(-)-OBn-NBoc-diazorhamidoic acid and S-(+)-OBn-NBoc-diazorhamidoic acid.

Figure 2 is a chromatogram of the optical isomers R-(-)-Aza-PBHA and S-(+)-Aza-PBHA.

Figure 3 is a chromatogram of the separation of (±)-Aza-PBHA by semi-preparative high performance liquid chromatography.

Figure 4 is a H ¹ -NMR spectrum of R-(-)-Aza-PBHA.

Figure 5 is a H ¹ -NMR spectrum of S-(+)-Aza-PBHA.

Figure 6 shows the effect of (±)-Aza-PBHA (A), R-(-)-Aza-PBHA (B) and S-(+)-Aza-PBHA (C) on the survival rate of IEC-6 small intestinal cells.

Figure 7 shows the Western blotting test for R-(-)-Aza-PBHA (3-10 μm), S-(+)-Aza-PBHA (3-10 μm) and (±)-Aza-PBHA (10 μm) for the small intestine The degree of expression of intracellular acetyl-Histone 3 protein.

The scope of the present invention comprises: (1) a pharmaceutical composition comprising at least one compound of the present invention and a pharmaceutically acceptable carrier, and (2) a method for treating mucosal ulcer or cancer by administering the same The therapeutically effective amount of the compound is administered to a subject.

"Treatment" as used herein refers to the administration of an active compound to an individual suffering from mucosal ulcers or cancer, or to an individual having symptoms or predispositions, the purpose of which is to cure, heal, slow, or lick the discomfort, symptoms or tendency of mucosal ulcers or cancer. Slow, change, correct, improve, enhance, influence, or reduce risk. By "effective dose" is meant the amount of active compound required to achieve the desired therapeutic effect on an individual. The effective dose may vary, as will be appreciated by those skilled in the art, depending on the route of administration, the application of the excipient, and the possible combination with other agents.

Mucositis is an inflammation and ulceration of the mucosa of the digestive tract. It is usually a side effect of cancer chemotherapy and radiation therapy, which may occur in the mouth or gastrointestinal tract.

Cancer contains solid tumors and blood tumors of different organs. Solid tumors include pancreatic cancer, gallbladder cancer, intestinal cancer, breast cancer and metastatic breast cancer, prostate cancer (including male hormone-dependent and independent prostate cancer), liver cancer, and lung cancer (eg, non-small cell lung cancer) (NSCLC), bronchioles and alveolar carcinoma (BAC) and lung adenocarcinoma), ovarian cancer (eg, progressive epithelial cancer and primary peritoneal cancer), Neck cancer, stomach cancer, esophageal cancer, head and neck cancer (such as head and neck squamous cell carcinoma), melanoma, neuroendocrine cancer (including metastatic neuroendocrine cancer), brain cancer (including neuroglioma, degenerative oligodendrocyte) Tumor, adult polymorphic glioma and adult degenerative astrocytoma), myeloma, and soft tissue sarcoma. Hematological tumors include acute myeloid leukemia (AML); chronic myelogenous leukemia (CML), including accelerated phase CML and CML acute conversion (CML-BP); acute lymphocytic leukemia (ALL); chronic lymphocytic leukemia (CLL) Hodgkin's disease (HD); non-Hodgkin's lymphoma (NHL), including follicular lymphoma and mantle cell lymphoma), B-cell lymphoma, T-cell lymphoma, multiple Myeloma (MM), Waldenstrom's macroglobulinemia, myelodysplastic syndrome (MDS), including persistent anemia (RA), ring-shaped lateral bud cell-resistant anemia (RARS), excessive bud cell-resistant anemia (RAEB) , and excessive bud cell refractory anemia and acute transformation (RAEB-T); and myeloproliferative syndromes.

The method of the present invention is carried out by administering the above-mentioned pharmaceutical composition by oral administration, or by parenteral administration, inhalation spray, area application, rectal suppository, nasal inhalation, oral injection, vaginal administration, or via implantable type. Storage tank. "Parenteral administration" refers to the route of administration through subcutaneous, intradermal, intravenous, muscular, joint, arterial, synovial fluid, sternum, intraspinal, intralesional, and intracranial injections or The method of administration of runoff.

Sterile injectable compositions, e.g., sterile aqueous or aqueous suspensions for intravenous injection, may be prepared in the form of suitable dispersing or wetting agents (e.g., Tween 80) and suspending agents which are known in the art. The preparation of the sterile injectable preparation may be a non-toxic, parenteral administration of a usable diluent or solvent (for example) A sterile injectable solution or suspension, such as 1,3-butanediol. Useful solvents or solvents include mannitol, water, Ringer's solution, and isotonic saline solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspension medium (for example, synthetic mono or diglycerides). Fatty acids, such as oleic acid and its glyceride derivatives, are useful in the preparation of sterile injectables, such as natural pharmaceutically acceptable oils such as olive oil, castor oil, and the like, especially their polyoxyethylated derivatives. The oil solution and suspension may be obtained by diluting or dispersing a long-chain alcohol or by diluting or dispersing carboxymethylcellulose or other dispersing agents. Other commonly used surfactants such as Tween or Spans or other similar emulsifiers or accelerators commonly used to make bioavailability of pharmaceutically acceptable solid, liquid or other dosage forms can also be used on the formulation.

The orally administered composition may be in any orally acceptable dosage form, including capsules, troches, emulsions, aqueous suspensions, dispersions, solutions, and the like. A commonly used carrier for tablets for oral use comprises lactose and corn starch. Lubricants, such as magnesium stearate, are typical additives. In oral capsules, the usual diluents include lactose and dried cornstarch. When a suspension or emulsion is administered orally, the active ingredient can be suspended or dissolved in the oily phase, together with emulsifying or suspending agents. Specific sweeteners, flavoring agents, or coloring agents can be added under certain requirements. Nasal or buccal inhalation spray ingredients can be prepared by methods known in the art of pharmaceutical formulations. The compounds of the present invention can also be used in the form of enteric plugs.

The pharmaceutically acceptable carrier must be "acceptable", including the necessity of coexistence with the active ingredient in a particular dosage form (preferably capable of stabilizing the active ingredient), while not causing harm to the individual upon administration. One or more stabilizers (eg, cyclodextrins) capable of forming a more soluble complex with the compounds of the present invention, Used as a pharmaceutical carrier to deliver an effective compound. Other carriers include cerium oxide sol, magnesium stearate, sodium lauryl sulfate, and D&C Yellow No. 10.

Suitable in vitro assays can be used to initially assess the effectiveness of the compounds of the invention in inhibiting tissue protein deacetylase. Effective compounds can further be tested for their effectiveness in treating mucosal ulcers or cancer in vitro or in vivo. For example, a compound can be administered in an animal model with mucosal ulcers or cancer (e.g., in a mouse experiment), and his therapeutic effect can be evaluated. Based on the experimental results, a suitable dosage range and route of administration can be determined.

The invention discloses: (1) separating the racemic mixture (±)-OBn-NBoc by using R-(-)-phenylethylamine (R-(-)-phenylethylamine) and S-(+)-phenethylamine, respectively. - Azatyrosine (structure is as follows), R-(-)-OBn-NBoc-diazorutamide with optical purity (enantiomeric excesses, ee%) > 99.90 (Table 1) and optical purity ee %>98.30 of S-(+)-OBn-NBoc-diazorudronic acid (Table 2); the novelty of this segmentation method includes increasing the optical segmentation efficiency by slow cooling crystallization, controlling the stirring rate and time for the partition Interacting with the partitioning reagent for a long time, effectively utilizing the difference in solubility of the formed salt to improve the separation efficiency of the optical substance; (2) separating the (±)-Aza-PBHA (structure as follows) to produce a novel compound by optical column separation R-(-)-Aza-PBHA and S-(+)-Aza-PBHA, and they all have the activity of inhibiting histone deacetylase, wherein (±)-Aza-PBHA has a geometric center, The optical rotation ~0 proves that its R/S mirror isomer is present in a ratio of 1:1.

The above list is considered to be sufficient to illustrate the invention without further elaboration. The following examples are for illustrative purposes only and are not limiting as to what is disclosed elsewhere. The published literature cited herein is incorporated herein by reference in its entirety.

Example 1: Intermediate racemic compound (±)-OBn-NBoc-diazorudronic acid (scientific name (±) 3-(5-(benzyloxy)pyridin-2-yl)-2-(t-butoxycarbonylamino) Establishment of analytical conditions for (±)benzyl-(5-(benzyloxy)pyridine-2-yl)-2-(tert-butoxycarbonylamino)propanoic acid

Intermediate racemic compound (±)-OBn-NBoc-diazorhamidoic acid in analytical high performance liquid chromatography system (Shimadzu LC-20AT, SIL-20A, SPD-M20A) with DAICEL CHIRALPAK® ID HPLC Analytical column (5 μm, 250 mm x 4.6 mm) was used to analyze the optical purity of the optically segmented product, and the reaction phase was analyzed by moving the phase n-hexane:ethanol:trifluoroacetic acid=80:20:0.1, and flowing at a flow rate of 1.0 mL/min for 20 minutes. The peak retention time of 10.21 minutes and 11.65 minutes was separately dissolved in CH ₂ Cl ₂ , and an equivalent TFA excision-Boc protecting group was added, followed by hydrogenation of the -OBn protecting group to obtain two diazon ruthenium optical isomers. The solution was dissolved in 1N hydrochloric acid, and the optical rotation was measured to obtain a specific optical rotation value of [α _D ] = -60.4 and [α _D ] = +58.6. Alignment literature [S-(+)-diazonium ursolic acid ratio [α _D ]=+59.3], estimated residence time 10.21 minutes for R-(-)-OBn-NBoc-diazoxamine Acid, residence time of 11.65 minutes is (S-(+)-OBn-NBoc-diazorudosamine (Fig. 1). Then used to determine the optical isomer purity when using this condition as an optical isomer. conditions of.

Preparation of (R)-(-)-3-(5-(benzyloxy)pyridin-2-yl)-2-(t-butoxycarbonylamino)propionic acid (R-(-)-OBn-NBoc-heavy Nitrosamine

Step: The racemic mixture (4±g of (±)-OBn-NBoc-diazonium bromate was dissolved in 500 mL of acetonitrile (ACN) and heated to complete dissolution. The insoluble microsolids were filtered by gravity and the filtrate continued. The heat was kept completely dissolved, and an equivalent amount of R-(+)-phenethylamine was added. After 5 minutes, the heating was stopped and the temperature was slowly returned to room temperature, and stirring was continued until the solid was completely precipitated. After 12 hours, the solid product was filtered to partially solidify Dissolve in dichloromethane (CH ₂ Cl ₂ ), extract with an equivalent amount of 1M H ₃ PO ₄ aqueous solution, the aqueous layer has a pH of about 5-6, dry the organic layer and remove the solvent with a vacuum cleaner to obtain a white solid. The mixture was used for optical quantitative analysis. The filtered solid was recrystallized five times in the same procedure (5 times of crystallization R-(-)-, S-(+)-separation ratio change was as shown in Table 1, yielding 17.15 g of white solid product, Analysis showed R-(-)-OBn-NBoc-diazorudronic acid with an enantiomeric excesses (ee%) > 99.90, yield 43%.

Preparation of (S)-(+)-3-(5-(benzyloxy)pyridin-2-yl)-2-(t-butoxycarbonylamino)propionic acid (S-(+)-OBn-NBoc-heavy Nitrosamine

30 g of the racemic mixture (±)-OBn-NBoc-diazologamine was dissolved in 500 mL of acetonitrile and heated to complete dissolution. The insoluble solids were filtered by gravitation and the filtrate was continuously heated to maintain complete dissolution. Equivalent S-(+)-phenethylamine was added, stirred for 10 minutes, cooled to room temperature, and then moved to -20 ° C for 24 minutes. After the solids were completely precipitated, the solid product was collected by filtration. A part of the solid was dissolved in dichloromethane (CH ₂ Cl ₂ ), extracted with an equivalent of 1 M H ₃ PO ₄ aqueous solution, the aqueous layer pH was about 5-6, the organic layer was dried and solvent was removed with a vacuum cleaner. A white solid mixture was obtained for optical quantitative analysis. The filtered solid was recrystallized 10 times in the same procedure (crystallization process R-, S-separation ratio changed as in Table 2) to obtain 4.62 g of a white solid product, which showed an optical purity (enantiomeric excesses, ee%) > 98.36. S-(+)-OBn-NBoc-diazorudronic acid, yield 15%.

Example 2: Establishment of novel compounds R-(-)-Aza-PBHA and S-(+)-Aza-PBHA [scientific name (R)-(-)- and S-(+)-t-butyl 3-(5) -(benzyloxy)pyridin-2-yl)-1-(4-(4-(hydroxyamino)-4-oxobutyl)anilino)-1-oxopropyl-2-ylcarbamate (R -(-)-and S-(+)-tn-butyl(3-(5-(benzyloxy)pyridin-2-yl)-1- ((4-(5-(hydroxyamino)-4,5-dioxopentyl)phenyl)amino)-1-oxopropan-2-yl)carbamate)]

The invention first establishes the analysis conditions of R-(-)-Aza-PBHA and S-(+)-Aza-PBHA by an analytical high performance liquid chromatography. The analytical column was an immobilized CHIRALPAK® ID HPLC analytical column (5 μm, 250 mm x 4.6 mm); the mobile phase was methanol: n-hexane: ethanol: diethanolamine (DEA) = 20:20:60 : 0.1; flow rate was 1 ml/min; injection amount was 20 μL; detection wavelength: 250 nm. Quantitative analysis found that the ratio of R-(-)-Aza-PBHA (appearing at 10.2 minutes) and S-(+)-Aza-PBHA (appearing at 11.65 minutes) in the racemic mixture (±)-Aza-PBHA It was 1 and 0.93, and its purity and structure were identified by an optical lucometer and a nuclear magnetic resonance instrument. Under this condition, the semi-prepared column and the best mobile phase were searched for optical separation of (±)-Aza-PBHA (Fig. 2).

Preparation of R-(-)- or S-(+)- by intermediate R-(-)-OBn-NBoc-diazorhamidoic acid or S-(+)-OBn-NBoc-diazorutanoic acid, respectively The process of pure optical isomers of Aza-PBHA still has some racemization, so an optical separation of (±)-Aza-PBHA is required. The present invention uses a preparative high performance liquid chromatography to separate R-(-)-Aza-PBHA and S-(+)-Aza-PBHA [Franco p., et al., Methods Mol. Biol. 2013, 970, 113- 126; Bruno G., et al. J. Chromatogr. A, 2014, 1339, 210-213]. Sample (±)-Aza-PBHA (dissolved in DMSO: MeOH = 1:4 concentration 10 mg / 1 mL, injection volume 500 μL with D2000 liquid chromatography (pump L-2130, UV-VIS detector L-2420, Hitachi) High-Technologies Corporation); the chromatographic column is Chiralpak® AD HPLC semipreparative columns 5 μm; detection wavelength is 250 nm; mobile phase is ACN: H ₂ O: DEA=70:30:0.1; flow rate Separation was carried out at 2 mL/min; flow washing for 50 minutes. R-(-)-Aza-PBHA appeared 16.4 minutes and S-(+)-Aza-PBHA appeared in 35.6 minutes (Fig. 3). After concentration and drying under reduced pressure, the purity of the product was identified by analytical grade high performance liquid chromatography, and the optical purity of R-(-)-Aza-PBHA was confirmed to be -12 (n=3) by an optical spectrophotometer at 25 °C. S-(+)-Aza-PBHA is +12 (n=3); the nuclear magnetic resonance spectrum also supports this structure (Fig. 4 & Fig. 5), showing that the analysis conditions are simple and feasible, and are applied to separation (±)- Aza-PBHA A process for the preparation of pure optical isomers.

(i) (±)-4-(4-(3-(5-(Benzyloxy)pyridin-2-yl)-2-(t-butoxycarbonylamino)propanamine)phenyl)butyrate Preparation of ester

Methyl 4-(4-phenylamine)butanoate (1.24 g, 6.42 mmol), (±)-3-(5-(benzyloxy)pyridin-2-yl)-2-(t-butoxycarbonyl) Amino)propionic acid ((±)-OBn-NBoc-diazorutamide) (2.17 g, 5.84 mmol) and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride The compound (1.12 g, 5.84 mmol) was dissolved in EtOAc (EtOAc m. The mixture was washed with EtOAc (EtOAc m. Melting point: 166-167 ° C; (400 MHz in CDCl ₃ , Bruker AVANCE-400): δ 1.31 ( s , 9H, Boc), 1.93 ( m , 2H, CH ₂ ), 2.30 (t, 2H, CH ₂ -CH ₂ -COOMe), 2.59 (t, 2H, CH ₂ ), 3.22 ( m 2H, CH ₂ ), 3.66 ( s , 3H, OMe), 4.43 ( t 1H CH) 5.13 ( s 2H OBn ), 6.97 ( d , 2H , J = 7.9 Hz, Ar-H), 7.09 ( d , 1H, J = 8.0 Hz, Pyr-H), 7.20 to 7.48 ( m , 9H Ar-H), 8.26 ( br , 1H, NH), 9.91 ( Br , 1H, NH).

(ii) Synthesis of (±)-4-(4-(3-(5-(benzyloxy)pyridin-2-yl)-2(t-butoxycarbonylamino)propanamine)phenyl)butyric acid

(±)-4-(4-(3-(5-(Benzyloxy)pyridin-2-yl)-2-(t-butoxycarbonylamino)propanamine)Phenyl)butyrate methyl ester (0.59 g, 1.07 mmol) and lithium hydroxide (0.26 g, 10.7 mmol) were dissolved in 10 mL of tetrahydrofuran / water (1:1) and heated at 60 ° C for 2 hours. The dropping was acidified to pH = 2 and extracted with dichloromethane. Dichloromethane was separated, then dried over magnesium sulfate to remove water, then concentrated in vacuo to give the title compound (0.51g, 90%) :( 400MHz in CDCl 3, Bruker AVANCE-400): δ 1.30 (s, 9H, Boc) , 1.74 ( m , 2H, CH ₂ ), 2.17 ( t , 2H, CH ₂ -CH ₂ -COOMe, J = 7.34 Hz), 2.52 ( t , 2H, CH ₂ , J = 7.55 Hz), 2.92-3.07 ( m , 2H, CH ₂ ), 4.44 ( m , 1H CH), 5.13 ( s , 2H OBn ), 7.09 ( d , 1H, J = 8.47 Hz, Ar-H), 7.21 (d, 1H, Ar-H, J = 8.52 Hz), 7.31 to 7.48 ( m , 9H Ar-H), 8.26 ( br , 1H, NH), 9.91 ( br , 1H, OH).

(iii) (±)-Tertibutyl 3-(5-(benzyloxy)pyridin-2-yl)-1-(4-(4-(hydroxyamino)-4-oxobutyl)anilino)- Synthesis of 1-oxopropyl-2-ylcarbamate Preparation of ((±)-Aza-PBHA)

(±)-4-(4-(3-(5-(Benzyloxy)pyridin-2-yl)-2(t-butoxycarbonylamino)propanamine)phenyl)butanoic acid (0.43 g, After dissolving in anhydrous tetrahydrofuran, ethyl chloroformate (0.1 mL, 0.88 mmol) and triethylamine (0.15 mL, 0.96 mmol) were added at 0 °C. Stir at 0 ° C for 30 minutes. After the insoluble matter was removed by filtration, the filtrate was added to a hydroxylamine methanol solution. The hydroxylamine methanol solution was prepared in situ, and a solution of hydroxylamine hydrochloride (0.11 g, 1.60 mmol) in methanol (1.1 mL) was added to a stirred solution of KOH (0.09 g, 1.60 mmol) in methanol (0.5 mL) at 0 °. . After stirring for 15 minutes to precipitate removed and the filtrate was stirred for 30 min (TLC monitoring) at room temperature, the solvent was removed in vacuo, the residue was extracted with EA / H ₂ O, and then in methanol \ recrystallized ether to give the subject Compound 3 (0.3 g, 74%). Melting point: >300 ° C; NMR (400 MHz in CDCl ₃ , Bruker AVANCE-400): δ 1.23 (s, 9H, Boc), 1.75 ( m , 2H, CH ₂ ), 1.93 ( t , 2H, CH ₂ -CH ₂ -CONHOH, J = 7.0 Hz), 2.51 ( t , 2H, CH ₂ , J = 6.5 Hz), 2.95 (m, 2H, CH), 4.45 ( m , 1H, CH), 5.14 ( s , 2H, OBn) , 6.97 ( d , 1H, J = 8.0 Hz, Ar-H), 7.09 ( d , 2H, J = 8.5 Hz), 7.21 ( d , 1H, J = 8.5 Hz, Ar-H), 7.32 - 7.47 ( m , 8H, Ar-H), 8.26 ( br , 1H, NH), 8.62 ( br , 1H, NH), 9.91 ( br , 1H, NH), 10.32 ( br , 1H, OH).

Example 3: Biological activity Effect of R-(-)-Aza-PBHA and S-(+)-Aza-PBHA on the survival rate of small intestine cells

MTT analysis showed that (±)-Aza-PBHA, R-(-)-Aza-PBHA and S-(+)-Aza-PBHA had no effect on the survival rate of IEC-6 small intestinal cells at a concentration of 10 μM or less, only S-( +)-Aza-PBHA partially affects cell viability at high concentrations of 20 μM (Figure 6)

R-(-)-Aza-PBHA, S-(+)-Aza-PBHA inhibit the activity of pan-HDAC

Western blotting method can be used to predict the degree of inhibition of pan-HDAC activity by observing the degree of expression of acetyl-histone 3 in small intestinal cells. The results showed that R-(-)-Aza-PBHA (3-10μM) and S-(+)-Aza-PBHA (3-10μM) significantly increased the effect of acetylated tissue protein on (±)-Aza-PBHA. (10 μM) was equivalent, and it was confirmed that both R-(-)-Aza-PBHA and S-(+)-Aza-PBHA inhibited the activity of pan-HDAC (Fig. 7).

Claims (4)

a compound having the following structural formula, A use of a compound as described in claim 1 for the preparation of a medicament for the treatment of mucosal ulcers. A use of a compound as described in claim 1 for the preparation of a medicament for the treatment of cancer. A third-butyl 3-(5-(benzyloxy)pyridin-2-yl)-1-(4-(4-(hydroxylamino)-4-oxobutyl)anilino)-1-oxopropyl group A racemic mixture of (-2-)-aminocarbamate ((±)Aza-PBHA) is isolated from a compound having a compound as described in claim 1, comprising a racemization of (±) Aza-PBHA The solution of the mixture was introduced into a high performance liquid chromatography (HPLC) column for flow washing.