KR20130122361A - 4-(benzimidazol-2-ylamino)pyrrolidine derivatives inhibiting beta-secretase's activity and pharmaceutical composition containing the same as an active ingredient - Google Patents

Abstract

The present invention relates to 4-(benzimidazole-2-ylamino)pyrrolidine derivatives of chemical formula 1 or pharmaceutically acceptable salts thereof; a pharmaceutical composition containing the same as an active ingredient for preventing and treating neurodegenerative diseases; and an inhibitor composition of the activation of beta-secretase (BACE), containing the same as an active ingredient. The 4-(benzimidazole-2-ylamino)pyrrolidine derivatives according to the present invention suppresses the activation of BACE and generation of beta-amyloid proteins which cause damage to nerve cells, thereby effectively preventing or treating neurodegenerative diseases such as Alzheimer’s disease and Down syndrome.

Description

4- (benzimidazol-2-ylamino) pyrrolidine derivatives that inhibit beta-secretase activity and pharmaceutical compositions containing them as active ingredients {4- (Benzimidazol-2-ylamino) pyrrolidine derivatives inhibiting beta- secretase's activity and pharmaceutical composition containing the same as an active ingredient}

The present invention provides 4- (benzimidazol-2-ylamino) pyrrolidine derivatives or pharmaceutically acceptable salts thereof that inhibit the activity of beta-secretase (β-secretase, hereinafter referred to as BACE), It relates to a pharmaceutical composition for the prevention and treatment of neurodegenerative diseases containing as an active ingredient and to an activity inhibitor composition of beta-secretase (BACE) containing the same as an active ingredient.

Alzheimer's disease (AD) is a degenerative brain disease that is closely related to aging characterized by loss of memory, cognition, reasoning, judgment, and coping due to neuronal damage. Pathological characteristics of Alzheimer's disease include intracellular accumulation of neurofibrous bundles in the areas related to cognitive activity of the brain and extracellular deposition of senile plaques consisting of Aβ protein as a major component. Aβ proteins consisting of 39-43 amino acids are known to exhibit neuronal toxicity. A number of reports indicate that Aβ is a pathological feature of Alzheimer's disease and is recognized as a major cause of disease development. Aβ peptides are produced by the hydrolysis of amyloid precursor protein (hereinafter referred to as APP), and a number of Aβ consisting of 39-43 amino acids are known. Aβ is produced by a pathway where APP is first cleaved at the N-terminus of the amyloid precursor protein (APP) by BACE and then cleaved at the C-terminus by gamma-secretase.

Thus, a beta-secretase enzyme, also named BACE, Asp or Memapsin ((a) Tang, J. et al., Proc . Natl . Acad . Sci . USA 2000 , 97 , 1456. ( b) Hussain, I. et al., Mol. Cell Neurosci. 1999 , 14 , 419. (c) Yan, R. et al., Nature 1999 , 402 , 533. (d) .Sinha, S, et al. , Nature 1999 , 402 , 537. (e) Inhibitors that inhibit the activity of Vassar, R. et al., Science 1999 , 286 , 735.) are drugs that can prevent Alzheimer's disease and treat underlying causes or symptoms. As it is available, many pharmaceutical companies are currently working to develop new inhibitors.

However, there have been no reports of developing beta-secretase inhibitors having a pyrrolidine backbone and excellent ability to inhibit beta-secretase activity.

 Tang, J. et al., Proc. Natl. Acad. Sci. U. S. A. 2000, 97, 1456.  Hussain, I. et al., Mol. Cell Neurosci. 1999, 14, 419.  Yan, R. et al., Nature 1999, 402, 533.  Sinha, S, et al., Nature 1999, 402, 537.  Vassar, R. et al., Science 1999, 286, 735.

It is an object of the present invention to provide novel 4- (benzimidazol-2-ylamino) pyrrolidine derivatives or pharmaceutically acceptable salts thereof which inhibit the activity of beta-secretase (BACE).

Another object of the present invention to provide a pharmaceutical composition for the prevention and treatment of neurodegenerative diseases containing the 4- (benzimidazol-2-ylamino) pyrrolidine derivative or a pharmaceutically acceptable salt thereof as an active ingredient. It is.

Another object of the present invention is to provide an active inhibitor composition of beta-secretase (BACE) containing the 4- (benzimidazol-2-ylamino) pyrrolidine derivative or a pharmaceutically acceptable salt thereof as an active ingredient. To provide.

The present invention is a 4- (benzimidazol-2-ylamino) pyrrolidine derivative represented by the following formula (1) or a pharmaceutically acceptable salt thereof, a pharmaceutical for the prevention and treatment of neurodegenerative diseases containing the same as an active ingredient It relates to a composition and an activity inhibitor composition of beta-secretase (BACE) containing the same as an active ingredient.

[Formula 1]

Hereinafter, the present invention will be described in more detail.

The present invention provides a novel 4- (benzimidazol-2-ylamino) pyrrolidine derivative represented by the following formula (1) or a pharmaceutically acceptable salt thereof:

[Formula 1]

In Formula 1,

A is CH or N;

Y is a bond, -CH ₂ -or -C (= 0)-;

R ₁ is halogen, (C1-C10) alkyl, (C1-C10) alkoxy, (C6-C12) aryl, (C3-C12) heteroaryl, (C6-C12) aryloxy, NR ₁₁ R ₁₂ , 5-membered to 7-membered cyclicamino or 5- to 7-membered heterocyclic amino comprising at least one heteroatom selected from nitrogen, oxygen and sulfur;

R ₁₁ and R ₁₂ are each independently (C 1 -C 10) alkyl or (C 6 -C 12) aryl;

R ₂ is (C 1 -C 10) alkyl, (C 1 -C 10) alkoxy or halogen;

R ₃ is hydrogen, (C 1 -C 10) alkyl,-(CH ₂ ) _m -COR ₄ ,-(CH ₂ ) _m -COOR ₄ ,-(CH ₂ ) _m -CONR ₄ R ₅ or -SO ₂ R ₄ ;

R ₄ and R ₅ are each independently hydrogen, (C 1 -C 10) alkyl or (C 6 -C 12) aryl;

m is an integer from 0 to 5;

Alkyl of said R _1, alkoxy, aryl or aryloxy, the alkyl or alkoxy group of R _2, the R ₃ alkyl and R _4, R _5, alkyl or aryl of R ₁₁ and R ₁₂ are halogen, alkyl, aryl, and It may be further substituted with one or more substituents selected from the group consisting of hydroxy.

In Formula 1, preferably R ₁ is (C1-C10) alkyl, (C6-C12) aryl, halogen-substituted (C6-C12) aryl, (C6-C12) aryloxy, di (C1-C10) alkyl Amino, 5- to 7-membered cyclicamino or 5- to 7-membered heterocyclic amino; R ₂ is (C 1 -C 10) alkyl, (C 1 -C 10) alkoxy or halogen; R ₃ is hydrogen, hydroxy (C 1 -C 10) alkyl, —COR ₄ , —CH ₂ COOR ₄ , —COOR ₄ , —CH ₂ CONR ₄ R ₅ or —SO ₂ R ₄ ; R ₄ and R ₅ are each independently hydrogen, (C 1 -C 10) alkyl or (C 6 -C 12) aryl.

,

,

,

,

,

또는

이다.In Formula 1, more preferably R ₁ is methyl, n-propyl, i-propyl, phenyl, phenyl substituted with fluorine, phenoxy, dimethylamino, piperidino or morpholino; R ₂ is methyl, methoxy, chloro or fluorine; R ₃ is hydrogen, hydroxyethyl,

,

,

,

,

,

or

to be.

More specifically exemplifying the 4- (benzimidazol-2-ylamino) pyrrolidine derivative represented by Formula 1 according to the present invention is as follows.

A simple preparation method of 4- (benzimidazol-2-ylamino) pyrrolidine derivative represented by Chemical Formula 1 and a pharmaceutically acceptable salt thereof according to the present invention is shown in the following Reaction Schemes 1, 2 and 3, Scheme 1 below is when Y in formula 1 is -C (= O)-, Scheme 2 is when Y in formula 1 is a bond, Scheme 3 corresponds to when Y in formula 1 is -CH _2- . . In Schemes 1 to 3 below, BOC is t-butoxycarbonyl, and A, R ₁ , R ₂ , and R ₃ are as defined in Chemical Formula 1.

[Reaction Scheme 1]

As shown in Scheme 1, when Y is -C (= O)-, the method for preparing 4- (benzimidazol-2-ylamino) pyrrolidine derivative is comprised of the following 5 to 7 steps. :

1) preparing the azido alcohol compound (B) by ring-opening an epoxide ring stereoselectively to the compound (A) by a known method;

2) reacting the hydroxyl group of the azido alcohol compound (B) with the carboxylic acid compound (C) using a known method to produce an azido carboxylate compound (D) ;

3) preparing an amino compound (E) by reducing the azido group of the azido carboxylate compound (D) using a known method;

4) preparing an isothiocyanate compound (F) by converting the amino group of the amino compound (E) into an isothiocyanate group by a known method;

5) converting the isothiocyanate group of the isothiocyanate compound (F) with a 1,2-phenylenediamine compound (G) to convert to a benzimidazole group to produce compound (1-1) ;

6) removing the Boc (t-butoxycarbonyl) group of the compound (1-1) with trifluoroacetic acid to obtain a compound (1-2) ; And

7) Compound (1-3) (R ₃ ≠ hydrogen) is reacted with known compound (1-2) by haloalkyl, haloaryl, haloalcohol, acid anhydride, carboxylic acid, carboxylic acid chloride, sulfonic acid chloride and the like. Getting steps.

[Reaction Scheme 2]

[In Reaction Scheme 2, X ₁ and X ₂ are each independently halogen; Z is nitrogen, oxygen or sulfur.]

As shown in Scheme 2, When Y is a bond, the method for preparing 4- (benzimidazol-2-ylamino) pyrrolidine derivative comprises the following 5 to 7 steps:

1) preparing an azido compound (I) by reacting the hydroxyl group of the azido alcohol compound (B) with a dihalo compound (H) using a known method;

2) preparing a compound (K) by reacting the halogen group of the azido compound (I) with a cyclic amine compound (J) using a known method;

3) preparing an amino compound (L) by reducing the azido group of the compound (K) by a known method;

4) preparing an isothiocyanate compound (M) by converting the amino group of the amino compound (L) into an isothiocyanate group by a known method;

5) converting the isothiocyanate group of the isothiocyanate compound ( M) with the 1,2-phenylenediamine compound (G) to convert to a benzimidazole group to produce compound (1-4) ;

6) removing Boc (t-butoxycarbonyl) group of compound (1-4) with trifluoroacetic acid to obtain compound (1-5) ; And

7) Compound (1-6) (R ₃ ≠ hydrogen) is reacted with known compound (1-5) by haloalkyl, haloaryl, haloalcohol, acid anhydride, carboxylic acid, carboxylic acid chloride, sulfonic acid chloride and the like. Getting steps.

Scheme 3

[In Reaction Scheme 3, X ₃ is halogen.]

As shown in Scheme 3, Y is -CH ₂ - is made to the case of 4-production method of (benz-ylamino imidazol-2-yl) pyrrolidine derivatives includes the steps 4 to 6 the process of:

1) preparing a compound (O) by reacting the hydroxy group of the azido alcohol compound (B) with a halomethyl compound (N) using a known method;

2) preparing an amino compound (P) by reducing the azido group of the compound (O) using a known method;

3) converting the amino group of the amino compound (P) to an isothiocyanate group by a known method to produce an isothiocyanate compound (Q) ;

4) converting the isothiocyanate group of the isothiocyanate compound (Q) with the 1,2-phenylenediamine compound (G) to convert to a benzimidazole group to produce compound (1-7) ;

5) removing Boc (t-butoxycarbonyl) group of compound (1-7) with trifluoroacetic acid to obtain compound (1-8) ; And

6) Compound (1-9) (R ₃ ≠ hydrogen) is reacted with known compound (1-8) by haloalkyl, haloaryl, haloalcohol, acid anhydride, carboxylic acid, carboxylic acid chloride, sulfonic acid chloride and the like. Getting steps.

The derivative of formula 1 of the present invention may be used in the form of a pharmaceutically acceptable salt, and salts include acid addition salts formed by pharmaceutically acceptable free acid. Acid addition salts include inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid or phosphorous acid and aliphatic mono and dicarboxylates, phenyl-substituted alkanoates, hydroxy alkanoates and alkanedioates From non-toxic organic acids such as aromatic acids, aliphatic and aromatic sulfonic acids. Such pharmaceutically innocuous salts include, but are not limited to, sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate chloride, bromide, Butyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, succinate, maleic anhydride, maleic anhydride, , Sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, Methoxybenzoate, phthalate, terephthalate, benzene sulfonate, toluene sulfonate, chlorobenzene sulfide Propyl sulphonate, naphthalene-1-yne, xylenesulfonate, phenylsulfate, phenylbutyrate, citrate, lactate,? -Hydroxybutyrate, glycolate, maleate, Sulfonate, naphthalene-2-sulfonate or mandelate.

On the other hand, 4- (benzimidazol-2-ylamino) pyrrolidine derivatives represented by the formula (1) according to the present invention has the effect of inhibiting the BACE activity has the effect of inhibiting the production of beta-amyloid protein . Accordingly, the present invention is effective in the treatment and prevention of Alzheimer's disease or similar Down syndrome disease caused by beta-amyloid protein. Accordingly, the present invention is a drug for preventing or treating neurodegenerative diseases in which 4- (benzimidazol-2-ylamino) pyrrolidine derivative represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof is contained as an active ingredient. To provide a pharmaceutical composition.

The present invention also relates to a beta-secretase (BACE) containing 4- (benzimidazol-2-ylamino) pyrrolidine derivative represented by Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient. An activity inhibitor composition is provided.

The dosage of 4- (benzimidazol-2-ylamino) pyrrolidine derivatives or pharmaceutically acceptable salts thereof according to the present invention to the human body is based on the age, weight, sex, dosage form, health condition and degree of disease of the patient. In general, based on an adult patient with a weight of 70 kg is 0.01 ~ 1000 ㎎ / day, depending on the doctor or pharmacist may be administered once a day or divided into several times at regular intervals have.

The pharmaceutical composition according to the present invention may be added to the 4- (benzimidazol-2-ylamino) pyrrolidine derivative or a pharmaceutically acceptable salt thereof by adding a conventional non-toxic pharmaceutically acceptable carrier, adjuvant and excipient. Oral administration such as tablets, capsules, troches, solutions, suspensions or the like, or intravenous, subcutaneous or intraperitoneal injections, creams, ointments or patches for topical administration. Or may be formulated into a parenteral preparation such as a visual route.

Oral administration of the pharmaceutical composition according to the present invention can be prepared in all the various forms, for example tablets, powders, dry syrups, chewable tablets, granules, chewing tablets, capsules, soft capsules, pills It can exist in many forms, such as drinks, sublingual tablets, etc. In the case of powder, the amount of the active ingredient is preferably 0.01 to 99.9% by weight, and the content is applied in a rational manner according to the formulation of the composition. The pharmaceutical composition according to the present invention may be difficult to maintain physical properties if the amount of the piperidine compound or pharmaceutically acceptable salt thereof according to the present invention exceeds the maximum total weight according to each formulation, and is less than the minimum weight. If the pharmacological effect of the active ingredient may not be sufficiently shown.

Tablets according to the present invention may be administered to a patient in any form or manner in which an effective amount is bioavailable, i. The form or manner can be easily selected and the composition according to the invention can comprise one or more pharmaceutically acceptable excipients, wherein the proportions and properties of such excipients are dependent on the solubility and chemical properties of the selected tablets, the chosen administration Determined by the route and standard pharmaceutical practice.

More specifically, the compositions according to the invention may comprise a therapeutically effective amount of the above-mentioned active ingredient as an essential ingredient together with one or more pharmaceutically acceptable excipients. Excipient materials can be solid or semisolid materials that can function as a vehicle or medium of the active ingredient, and suitable excipients are well known in the art. Excipient materials may be selected in connection with the intended dosage form, specifically for tablets, powders, chewable tablets, granules, chewing tablets, capsules, soft capsules, pills, sublingual tablets or syrups, The therapeutically active drug component may be combined with any oral nontoxic pharmaceutically acceptable inert excipient such as lactose or starch. Optionally, the pharmaceutical tablets of the present invention may be formulated with pharmaceutically acceptable additives such as binders such as amorphous cellulose, gum tragacanth or gelatin, disintegrants such as alginic acid, lubricants such as magnesium stearate, glidants such as colloidal silicon dioxide, Sweeteners such as oz or saccharin, coloring agents such as peppermint or methyl salicylate, or flavoring agents.

Because of their ease of administration, tablets may be the most advantageous oral unit dosage form, and if desired, tablets may be coated with sugar, shellac or other enteric coatings by standard aqueous or non-aqueous techniques, respectively. Tablets or capsules preferably contain about 0.1 mg to 100 mg of active ingredient.

As described above, the 4- (benzimidazol-2-ylamino) pyrrolidine derivative represented by Formula 1 according to the present invention is effective in the prevention and treatment of various diseases related thereto because of its excellent BACE inhibitory activity, It can be usefully used for the prevention or treatment of neurodegenerative diseases such as Alzheimer's and Down syndrome known to be related to BACE inhibitory activity.

Hereinafter, the present invention will be described in detail through Examples and Experimental Examples. However, the following Examples and Experimental Examples are merely illustrative of the present invention, and the content of the present invention is not limited by the following Examples.

[ Example  1] (3S, 4S) -4- (5- methyl -1H- benzo [d] imidazol-2- ylamino ) pyrrolidine -3- yl biphenyl -4- carboxylate  Preparation of (Compound 102)

Step 1 : Preparation of Compound B

Compound A (3.0 g, 16.2 mmol) was dissolved in ether (10 mL) by a known method (WO 2009/153554) and (R, R) -N, N' -bis (3,5-bis-tert-butylsally Silidene) -1,2-cyclohexanediaminochrome (III) chloride (0.2 g, 0.3 mmol) is added. After stirring the reaction mixture for 30 minutes, azidotrimethylsilane (3.2 mL, 24.2 mmol) is added and stirred at room temperature for 24 hours. The reaction was concentrated under reduced pressure, dissolved in methanol (50 mL), p-toluenesulfonic acid (1 g) was added thereto, and stirred for 30 minutes. Saturated aqueous sodium hydrogen carbonate solution (50 mL) was added to the reaction, followed by extraction with ethyl acetate (2 X 50 mL), followed by removal of water with sodium sulfate and concentration. The residue was separated by silica gel column chromatography (hexane: ethyl acetate = 2: 1) to give azido alcohol compound B as a yellow solid (3.0 g, 81%).

¹ H-NMR (300 MHz, CDCl 3) δ 4.25 (br s, 1H), 3.93 (br s, 1H), 3.55-3.75 (m, 2H), 3.30-3.50 (m, 2H), 2.05 (br s, 1H), 1.46 (s, 9H).

Step 2: Preparation of Compound D-1

Azido alcohol Compound B (221 mg, 0.97 mmol) was dissolved in dimethylformamide (6 mL), and then biphenyl-4-carboxylic acid (Compound C-1 , 230 mg, 1.16 mmol), 1- (3-dimethylamino Propyl) -3-ethylcarbodiimide (EDC; 223 mg, 1.16 mmol) and 4- N, N -dimethylaminopyridine (DMAP; 118 mg, 0.97 mmol) were added and stirred at room temperature for 24 hours. Saturated aqueous sodium hydrogen carbonate solution (20 mL) is added followed by extraction with ethyl acetate (2 x 20 mL). The organic layer was washed with 0.5N aqueous hydrochloric acid solution (20 mL) and distilled water (20 mL), and then dried over sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane: ethyl acetate = 5: 1) to give compound D-1 (360 mg, 91%).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 1.49 (s, 9H), 3.53-3.87 (m, 4H), 4.23 (m, 1H), 5.36 (m, 1H), 7.27-7.51 (m, 4H) 7.61-7.70 (m, 4H) 8.08 (d, J = 8.4 Hz, 2H).

Step 3: Preparation of Compound E-1

Compound D-1 (350 mg, 0.86 mmol) was dissolved in ethanol (10 mL), 10% Pd-C (25 mg) was added, followed by stirring for 12 hours under a hydrogen balloon. After filtration through a pad of celite and concentration, the residue was purified by silica gel column chromatography (hexane: ethyl acetate = 2: 1) to give compound E-1 (279 mg, 85%).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 1.49 (s, 9H), 1.50 (br s, 2H), 3.20-3.30 (m, 1H), 3.41-3.50 (m, 1H), 3.52-3.69 (m , 2H), 3.87 (dd, J = 12.6, 5.3 Hz, 1H), 5.14 (m, 1H), 7.40-7.51 (m, 4H) 7.61-7.70 (m, 4H) 8.09 (d, J = 8.4 Hz, 2H).

Step 4: Preparation of Compound F-1

Compound E-1 (270 mg, 0.71 mmol) was dissolved in dichloromethane (10 mL), and then di (2-pyridyl) thiocarbonate (DPT; 163 mg, 0.85 mmol) was added thereto. Stir at room temperature for 5 hours. The reaction mixture was concentrated and purified by silica gel column chromatography (hexane: ethyl acetate = 8: 1) to give compound F-1 (255 mg, 85%).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 1.49 (s, 9H), 3.21-3.32 (m, 1H), 3.42-3.53 (m, 1H), 3.54-3.69 (m, 2H), 3.90 (m, 1H), 5.14 (m, 1H), 7.41-7.52 (m, 4H) 7.60-7.72 (m, 4H) 8.09 (d, J = 8.4 Hz, 2H).

Step 5: Preparation of Compound 101

In a solution of Compound F-1 (250 mg, 0.59 mmol) in tetrahydrofuran (4 mL), 4-methyl-phenylene-1,2-diamine (Compound G-1 , 87 mg, 0.71 mmol) and mercury oxide (HgO; 367 mg, 0.86 mmol) was added thereto and stirred at 60 ° C for 12 hours. The reaction mixture was filtered through a pad of celite, distilled water (10 mL) was added, extracted with ethyl acetate (10 mL), water was removed with sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane: ethyl acetate = 1: 1) to give compound 101 (205 mg, 70%).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 8.15 (d, J = 6.9 Hz, 2H), 7.67 (d, J = 8.4 Hz, 2H), 7.53 (m, 3H), 7.27 (m, 1H), 6.90 (d, J = 7.8 Hz, 1H), 5.32 (s, 1H), 4.26 (s, 1H), 3.92-3.61 (m, 4H), 2.43 (s, 3H), 1.49 (m, 9H).

Step 6: Preparation of Compound 102

Compound 101 (90 mg, 0.18 mmol) was dissolved in dichloromethane (3 mL), trifluoroacetic acid (6 mL) was added, and the mixture was stirred at room temperature for 2 hours. The reaction mixture is concentrated under reduced pressure, neutralized with 1N aqueous sodium hydroxide solution, extracted with ethyl acetate (15 mL), washed with distilled water (20 mL), dried over sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane: methanol = 12: 1) to give compound 102 (70 mg, 95%).

¹ H NMR (300 MHz, CDCl ₃ ) δ 8.15 (m, 2H), 7.72-7.65 (m, 4H), 7.63-7.42 (m, 3H), 7.31-7.22 (m, 1H), 6.90 (m, 1H ), 5.25 (m, 1H), 4.07 (m, 1H), 3.55-3.33 (m, 2H), 3.30-3.26 (m, 2H), 2.43 (s, 3H).

[ Example  2] (3S, 4S) -1- acetyl -4- (5- methyl -1H- benzo [d] imidazol-2-ylamino) pyrrolidin-3-yl biphenyl -4- carboxylate  Preparation of (Compound 103)

Acetic anhydride (4 mg, 39 mmol) in a solution of 102 (13 mg, 31.5 mmol) and triethylamine (6 mL, 43 mmol) obtained in step 6 of Example 1 in anhydrous dichloromethane (2 mL). Was added and stirred at room temperature for 2 hours. The reaction was concentrated under reduced pressure and purified by silica gel column chromatography (dichloromethane: methanol = 30: 1) to give compound 103 (13 mg, 95%).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 8.19-8.09 (m, 3H), 7.69-7.62 (m, 4H), 7.50-7.41 (m, 3H), 7.32-7.23 (m, 2H), 6.91- 6.90 (m, 1H), 5.67-5.62 (m, 1H), 4.82-4.79 (m, 1H), 4.23-4.01 (m, 2H), 3.87-3.82 (m, 1H), 3.73-3.69 (m, 1H ), 2.77 (s, 3H), 2.41 (s, 3H).

[ Example  3] (3S, 4S) -1- benzoyl -4- (5- methyl -1H- benzo [d] imidazol-2-ylamino) pyrrolidin-3-yl biphenyl -4- carboxylate  Preparation of (Compound 104)

Compound 102 (13 mg, 31.5 mmol) obtained in Step 6 of Example 1 and benzoyl chloride were reacted in the same manner as in Example 2, obtaining Compound 104 (14 mg, 89%).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 13.0 (brs, 1H), 8.14 (d, J = 8.1 Hz, 1H), 8.09-8.06 (m 2H), 8.03-7.42 (m, 11H), 7.22- 6.95 (m, 2H), 6.58-6.53 (m, 1H), 5.70-5.68 (m, 1H), 4.88-4.66 (m, 1H), 4.22-4.01 (m, 2H), 3.82-3.56 (m, 2H ), 3.27 (S, 3 H).

[ Example  4] (3S, 4S) -1- (2- 메틸oxy -2- oksoethyl ) -4- (5- methyl -1H-benzo [d] imidazol-2-ylamino) pyrrolidin-3-yl biphenyl -4- carboxylate  Preparation of (Compound 105)

Compound 102 (13 mg, 31.5 mmol) obtained in step 6 of Example 1 and methyl bromoacetate were reacted in the same manner as in Example 2 to obtain compound 105 (11 mg, 75%).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 8.17 (d, J = 6.8 Hz, 2H), 7.72 (d, J = 6.8 Hz, 2H), 7.66-7.26 (m, 1H), 7.52-7.51 (m , 3H), 7.43-7.42 (m, 1H), 7.28-7.26 (m, 1H), 6.92-6.91 (m, 1H), 5.30-5.28 (m, 1H), 4.22-4.20 (m, 1H), 3.81 -3.79 (m, 3H), 3.79-3.74 (m, 2H), 3.60 (s, 2H), 3.35-3.32 (m, 1H), 3.15-3.13 (m, 1H), 2.97-2.95 (m, 1H) , 2.44 (s, 3 H).

[ Example  5] (3S, 4S)- tert - butyl  3- ( biphenylcarbonyloxy ) -4- (5- 메틸oxy Preparation of -1H-benzo [d] imidazol-2-ylamino) pyrrolidine-1-carboxylate (Compound 106)

Compound F-1 obtained from Step 4 of Example 1 and 4-methoxy-1,2-phenylenediamine (Compound G-2 ) were reacted in the same manner as Step 5 of Example 1 to obtain Compound 106 . .

¹ H-NMR (300 MHz, CDCl ₃ ) δ 8.17 (d, J = 8.1 Hz, 2H), 7.73 (d, J = 8.4 Hz, 2H), 7.52-7.43 (m, 3H), 7.27-7.26 (m , 1H), 6.97 (d, J = 2.4 Hz, 1H), 5.34 (s, 1H), 4.22 (s, 1H), 3.93-3.58 (m, 7H), 3.84 (s, 3H), 1.52 (m, 9H).

[ Example  6] (3S, 4S)- tert - butyl  3- ( biphenylcarbonyloxy ) -4- (5- fluoro Preparation of -1H-benzo [d] imidazol-2-ylamino) pyrrolidine-1-carboxylate (Compound 107)

Compound 107 was obtained by reacting Compound F-1 obtained with Step 4 of Example 1 with 4-fluorobenzene-1,2-diamine (Compound G-3 ) in the same manner as Step 5 of Example 1.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 10.62 (bs, 1H), 8.18-8.18 (m, 2H), 7.73 (d, J = 8.1 Hz, 2H), 7.65 (d, J = 8.1 Hz, 2H ), 7.52-7.43 (m, 3H), 7.29-7.28 (m, 1H), 7.10 (d, J = 8.7 Hz, 1H), 6.80 (bs, 1H), 6.80-6.78 (m, 1H), 5.32 ( s, 1H), 4.24 (s, 1H), 3.95-3.81 (m, 3H), 3.70-3.68 (m, 1H), 1.46 (m, 9H).

[ Example  7] (3S, 4S)- tert - butyl  3- ( biphenylcarbonyloxy ) -4- (5- chloro Preparation of -1H-benzo [d] imidazol-2-ylamino) pyrrolidine-1-carboxylate (Compound 108)

Compound F-1 and 4-chloro-1,2-phenylenediamine (Compound G-4 ) obtained from Step 4 of Example 1 were reacted in the same manner as Step 5 of Example 1, to obtain Compound 108 .

¹ H-NMR (300 MHz, CDCl ₃ ) δ 10.56 (s, 1H), 8.18 (d, J = 8.0 Hz, 2H), 7.74 (d, J = 8.3 Hz, 2H), 7.66 (d, J = 7.2 Hz, 2H), 7.54-7.38 (m, 3H), 7.31 (d, J = 8.3 Hz, 1H), 7.14-7.01 (m, 1H), 5.33 (s, 1H), 4.21 (s, 1H), 3.98 -3.56 (m, 4H), 1.52 (m, 9H).

[ Example  8] (3S, 4S)- tert - butyl  3- (4'- fluorobiphenylcarbonyloxy ) -4- (5-methyl-1H-benzo [d] imidazol-2-ylamino) pyrrolidine-1-carboxylate (Compound 109)

The azido alcohol compound B obtained in Step 1 of Example 1 was reacted with 4'-fluorobiphenyl-3-benzoic acid (Compound C-2 ) in the same manner as in Steps 2 to 5 of Example 1 to obtain a compound. 109 was obtained.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 8.26 (s, 1H), 8.06 (d, J = 7.5 Hz, 1H), 7.82 (d, J = 7.8, 1H), 7.64-7.52 (m, 3H) , 7.29-7.23 (m, 1H), 7.23-7.13 (m, 3H), 6.70-6.80 (m, 1H), 5.33 (s, 1H), 4.25 (s, 1H), 3.94-3.63 (m, 4H) , 2.43 (s, 3 H), 1.49 (d, J = 14.1 Hz, 9 H).

[ Example  9] (3S, 4S)- tert - butyl  3- (5- chloro -1H- benzo Preparation of [d] imidazol-2-ylamino) -4- (3-phenoxybenzoyloxy) pyrrolidine-1-carboxylate (Compound 110)

The azido alcohol compound B obtained in step 1 of Example 1 was reacted with 3-phenoxybenzoic acid (compound C-3 ) in the same manner as in steps 2 to 4 of Example 1, and 4-chloro-1, Compound 110 was obtained by reacting the same method as Step 5 of Example 1 using 2-phenylenediamine (Compound G-4 ).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 10.37 (s, 1H), 7.83 (d, J = 7.5 Hz, 1H), 7.73 (s, 1H), 7.51-7.34 (m, 3H), 7.52-7.34 (m, 4H), 7.33-7.15 (m, 3H), 7.11-6.97 (m, 3H), 5.32-5.25 (m, 1H), 4.21-4.11 (m, 1H), 3.95-3.55 (m, 4H) 1.50 (m, 9 H).

[ Example  10] (3S, 4S)- tert - butyl  3- (5- chloro -1H- benzo Preparation of [d] imidazol-2-ylamino) -4- (3-phenoxybenzoyloxy) pyrrolidine-1-carboxylate (Compound 111)

The azido alcohol compound B obtained in step 1 of Example 1 was reacted with 4-phenoxybenzoic acid (compound C-4 ) in the same manner as in steps 2 to 4 of Example 1, and 4-chloro-1, Compound 111 was obtained by reacting 2-phenylenediamine (Compound G-4 ) in the same manner as in Step 5 of Example 1.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 10.60 (s, 1H), 8.06 (d, J = 8.5 Hz, 1H), 7.43 (t, J = 7.87 Hz, 1H), 7.31-7.20 (m, 2H ) 7.13-7.00 (m, 5H), 5.27 (s, 1H), 4.19 (s, 1H), 3.94-3.58 (m, 4H), 1.49 (m, 9H).

[ Example  11] (3S, 4S) tert - butyl  3- (5- chloro -1H- benzo Preparation of [d] imidazol-2-ylamino) -4- (3-phenoxybenzoyloxy) pyrrolidine-1-carboxylate (Compound 112)

The azido alcohol compound B obtained in step 1 of Example 1 was reacted with 4-isopropylbenzoic acid (compound C-5 ) in the same manner as in steps 2 to 4 of Example 1, and 4-fluoro-1 Compound 112 was obtained by reacting in the same manner as in Step 5 of Example 1, using, 2-phenylenediamine (Compound G-3 ).

¹ H NMR (300 MHz, CDCl ₃ ) δ 10.62-10.51 (m, 1H) 8.03 (d, J = 7.1 Hz, 2H), 7.36 (d, J = 8.2 Hz, 2H), 7.28-7.23 (m, 1H) , 7.08 (d, J = 8.8 Hz, 1H), 6.83-6.76 (m, 1H), 5.28-5.27 (s, 1H), 4.21-4.17 (m, 1H), 3.92-3.86 (m, 3H), 3.69 -3.63 (m, 1H), 3.05-2.96 (m, 1H), 1.50 (m, 9H), 1.29 (d, J = 6.9 Hz, 6H).

[ Example  12] (3S, 4S) tert - butyl  3- (5- chloro -1H- benzo Preparation of [d] imidazol-2-ylamino) -4- (3-phenoxybenzoyloxy) pyrrolidine-1-carboxylate (Compound 113)

The azido alcohol compound B obtained in step 1 of Example 1 was reacted with 4- (dimethylamino) benzoic acid (compound C-6 ) in the same manner as in steps 2 to 4 of Example 1, and 4-fluoro Compound 113 was obtained by reacting the same method as Step 5 of Example 1 using -1,2-phenylenediamine (Compound G-3 ).

¹ H NMR (300 MHz, CDCl ₃ ) δ 10.86 (brs, 1H), 7.95 (d, J = 8.6 Hz, 2H), 7.30-7.27 (m, 1H), 7.11-7.08 (m, 1H), 6.82-6.76 (brs, 1H), 6.67 (d, J = 9.0 Hz, 2H), 5.22 (s, 1H), 4.17-4.11 (m, 1H), 3.89-3.57 (m, 4H), 3.09 (s, 6H), 1.50 (m, 9 H).

[ Example  13] (3S, 4S) tert - butyl  3- (5- chloro -1H- benzo Preparation of [d] imidazol-2-ylamino) -4- (3-phenoxybenzoyloxy) pyrrolidine-1-carboxylate (Compound 114)

To a solution of Compound 112 (140 mg, 0.28 mmol) obtained in Example 11 in dichloromethane (2 mL) was added trifluoroacetic acid (10 mL) and stirred at room temperature for 2 hours. The reaction mixture was concentrated, neutralized with saturated aqueous sodium hydrogen carbonate solution, extracted with ethyl acetate (20 mL), washed with distilled water (20 mL), dried over sodium sulfate, concentrated under reduced pressure to obtain a solid compound, and the solid compound was separated. Used for the next reaction without purification.

The solid compound (25 mg, 65 mmol) was dissolved in dichloromethane (5 mL), followed by diisopropylethylamine (12 mL) and propanesulfonyl chloride (0.1 M dichloromethane solution, 0.75 mL, at 0 ° C). 75 mmol) is added and stirred for 2 hours. The reaction solution was concentrated under reduced pressure and purified by silica gel column chromatography (dichloromethane: methanol = 9: 1) to give compound 114 (32 mg, 63%).

¹ H NMR (300 MHz, CDCl ₃ ) δ 8.01 (d, J = 8.2 Hz, 2H), 7.35 (d, J = 8.2 Hz, 2H), 7.27-7.22 (m, 1H), 7.09-7.06 (m, 1H ), 6.83-6.72 (m, 1H), 5.32-5.31 (m, 1H), 4.33 (d, J = 5.7 Hz, 1H), 3.99-3.85 (m, 3H), 3.65 (m, J = 11.4 Hz, 1H), 3.06-2.95 (m, 3H), 1.91-1.83 (m, 2H), 1.28 (d, J = 6.9 Hz, 6H), 1.03 (t, J = 7.4 Hz, 3H)

[ Example  14] (3S, 4S) tert - butyl  3- (5- chloro -1H- benzo Preparation of [d] imidazol-2-ylamino) -4- (3-phenoxybenzoyloxy) pyrrolidine-1-carboxylate (Compound 115)

Compound 115 was obtained by reacting the same method as in Example 13, except that benzenesulfonyl chloride was used instead of propanesulfonyl chloride from compound 112 (48 mg, 0.10 mmol) obtained in Example 11 (43 mg, 84%).

¹ H NMR (300 MHz, CDCl ₃ ) δ 7.90 (d, J = 7.6 Hz, 2H), 7.67-7.54 (m, 5H), 7.26-7.23 (m, 3H), 7.09-7.05 (m, 1H), 6.82 -6.76 (m, 1H), 5.23 (d, J = 3.9 Hz, 1H), 4.20 (d, J = 4.8 Hz, 1H), 3.91-3.85 (m, 1H), 3.70-3.57 (m, 3H), 3.03-2.94 (m, 1H), 1.29-1.25 (m, 6H)

[ Example  15] (3S, 4S) tert - butyl  3- (5- chloro -1H- benzo Preparation of [d] imidazol-2-ylamino) -4- (3-phenoxybenzoyloxy) pyrrolidine-1-carboxylate (Compound 116)

Compound 116 was reacted with the same method as in Example 13 from Compound 113 (24 mg, 0.05 mmol) obtained in Example 12. Obtained (8 mg, 31%).

¹ H NMR (300 MHz, DMSO-d ₆ ) δ 7.81 (d, J = 8.7 Hz, 2H), 7.34-7.29 (m, 1H), 7.14-7.10 (m, 1H), 7.00-6.96 (m, 1H) , 6.74-6.68 (m, 3H), 5.39 (brs, 1H), 4.42 (brs, 1H), 3.88-3.80 (m, 2H), 3.48-3.42 (m, 2H), 3.23-3.03 (m, 2H) , 3.0 (s, 6H), 1.73-1.65 (m, 2H), 0.92 (t, J = 7.4 Hz, 3H)

[ Example  16] (3S, 4S) tert - butyl  3- (5- chloro -1H- benzo Preparation of [d] imidazol-2-ylamino) -4- (3-phenoxybenzoyloxy) pyrrolidine-1-carboxylate (Compound 117)

Example 12 from Compound 113 (25 mg, 51.8 mmol) obtained by the reaction in the same manner as in Example 13 to give the compound 117 (6mg, 22%).

¹ H NMR (300 MHz, CDCl ₃ ) δ 7.91 (d, J = 7.8 Hz, 2H), 7.70-7.65 (m, 1H), 7.61-7.55 (m, 4H), 7.25-7.22 (m 1H), 7.07 ( dd, J = 2.3 Hz, 9.28 Hz, 1H), 6.82-6.76 (m, 1H), 6.57 (d, J = 9.0 Hz, 2H), 5.16 (d, J = 4.3 Hz, 1H), 4.13 (d, J = 5.3 Hz, 1H), 3.87-3.82 (m, 1H), 3.73-3.66 (m, 2H), 3.57-3.53 (m, 1H), 3.09 (s, 6H).

[ Example  17] (3S, 4S) -1- ( tert - butoxycarbonyl ) -4- (5- fluoro -1H-benzo [d] imidazol-2-ylamino) pyrrolidin-3-yl 6- methylnicotinate  Preparation of (Compound 118)

Step 1 : Preparation of Compound D-2

Azido alcohol compound B (221 mg, 0.97 mmol) obtained in step 1 of Example 1 was dissolved in dimethylformamide (6 mL), and 6-methylnicotinic acid (compound C-7 ) (159 mg, 1.16 mmol), 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide (EDC; 223 mg, 1.16 mmol), 4- N, N -dimethylaminopyridine (DMAP; 118 mg, 0.97 mmol) were added and room temperature Stir at 24 hours. To the reaction was added saturated aqueous sodium hydrogen carbonate solution (20 mL) and extracted with ethyl acetate (2 x 20 mL). The organic layer was washed with 0.5N aqueous hydrochloric acid solution (20 mL) and distilled water (20 mL), and then dried over sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane: ethyl acetate = 5: 1) to give compound D-2 (330 mg, 98%).

H NMR (300 MHz, CDCl ₃ ) δ 1.48 (9, 9H), 2.64 (s, 3H), 3.55-3.86 (m, 4H), 4.22 (m, 1H), 5.34 (m, 1H), 7.24-7.27 (m, 1 H), 8.14 (dd, J = 8.1, 2.1 Hz, 1 H), 9.07 (s, 1 H).

Step 2 : Preparation of Compound E-2

Compound D-2 (320 mg, 0.92 mmol) was dissolved in ethanol (10 mL), Pd-C (10%, 30 mg) was added, followed by stirring for 12 hours under a hydrogen balloon. After filtration through a pad of celite, the residue was purified by silica gel column chromatography (hexane: ethyl acetate = 2: 1) to give compound E-2 (193 mg, 65%).

H NMR (300 MHz, CDCl ₃ ) δ 1.17 (br s, 2H), 1.48 (9, 9H), 2.64 (s, 3H), 3.29-3.34 (m, 1H), 3.48-3.50 (m, 1H), 3.65-3.73 (m, 2H), 3.91 (dd, J = 12.6, 5.1 Hz, 1H), 7.23-7.27 (m, 1H), 8.14 (dd, J = 8.1, 2.1 Hz, 1H), 9.07 (s, 1H).

Step 3 : Preparation of Compound F-2

Compound E-2 (190 mg, 0.59 mmol) was dissolved in dichloromethane (10 mL), and then di (2-pyridyl) thiocarbonate (DPT; 136 mg, 0.71 mmol) was added thereto. Stir at room temperature for 5 hours. The reaction mixture was concentrated and purified by silica gel column chromatography (hexane: ethyl acetate = 6: 1) to give compound F-2 (167 mg, 78%).

H NMR (300 MHz, CDCl ₃ ) δ 1.48 (9, 9H), 2.64 (s, 3H), 3.29-3.34 (m, 1H), 3.48-3.50 (m, 1H), 3.65-3.73 (m, 2H) , 4.30 (s, 1 H) 5.48 (m, 1 H), 7.25-7.27 (m, 1 H), 8.12 (d J = 8.1 Hz, 1 H), 9.05 (s, 1 H).

Step 4 : Preparation of Compound 118

4-fluoro-1,2-phenylenediamine (Compound G-3 ) (50 mg, 0.39 mmol) in a solution of Compound F-2 (120 mg, 0.33 mmol) in tetrahydrofuran (2 mL), Add mercury oxide (HgO; 153 mg, 0.36 mmol) and stir at 60 ° C for 12 hours. The reaction mixture is filtered through a pad of celite, distilled water (10 mL) is added and extracted with EtOAc (10 mL). Water is removed with sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane: ethyl acetate = 1: 1) to give compound 118 (107 mg, 71%).

¹ H NMR (300 MHz, CDCl ₃ ) δ 10.3 (brs, 1H), 9.17 (s, 1H), 8.23 (d, J = 7.9 Hz, 2H), 7.31 (d, J = 8.2 Hz, 1H), 7.26 ( t, 1H), 7.08 (d, J = 9.5 Hz, 1H) 6.80 (brs, 1H), 5.33 (s, 1H), 4.24 (s, 1H), 3.92-3.63 (m, 4H), 2.68 (s, 3H), 1.49 (s, 9H)

[ Example  18] (3S, 4S) tert - butyl  3- (5- fluoro -1H- benzo Preparation of [d] imidazol-2-ylamino) -4- (4- (piperidin-1-yl) phenoxy) pyrrolidine-1-carboxylate (Compound 119)

Step 1: Preparation of Compound I-1

Azido alcohol compound B (1.0 g, 4.38 mmo) obtained from step 1 of Example 1 was added to a solution of toluene (6 mL) cesium carbonate (1.7 g, 5.25 mmol), 1,10-phenanthroline (240 mg, 1.31 mmol), 1,4-diiodobenzene (Compound H-1 ) (1.7 g, 5.25 mmol) were added, and residual gas was removed with argon. Then, copper iodide (CuI; 167 mg, 0.88 mmol) was added and During heating to 110 ° C. The reaction is filtered through a pad of celite and washed with ethyl acetate (20 mL). The organic layer is washed with distilled water (2 x 20 mL), dried over sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane: ethyl acetate = 7: 1) to give compound I-1 (1.2 g, 64%).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 1.47 (s, 9H), 3.48-3.78 (m, 2H), 4.13-4.15 (m, 1H), 4.62 (m, 1H), 6.66 (d, J = 7.4 Hz, 2H), 7.58 (d, J = 7.4 Hz, 2H).

Step 2: Preparation of Compound K-1

Compound I-1 (500 mg, 1.16 mmol) was dissolved in DMSO (1.5 mL), then piperidine (Compound J-1 , 175 mL, 1.74 mmol), L-proline (272 mg, 0.23 mmol), potassium carbonate (320 mg, 2.23 mmol) was added and residual gas was removed. Then, copper iodide (CuI; 22. 2 mg, 0.12 mmol) was added thereto and stirred at 80 ° C. for 24 hours. The reaction is filtered through a pad of celite and washed with ethyl acetate (20 mL). The organic layer is washed with distilled water (3 x 20 mL), dried over sodium sulfate and concentrated under reduced pressure. The residue was separated by silica gel column chromatography (hexane: ethyl acetate = 5: 1) to give compound K-1 (220 mg, 49%).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 1.47 (s, 9H), 1.46-1.57 (m 2H), 1.59-2.04 (m, 4H), 3.03-3.06 (m, 4H), 3.51-3.74 (m , 4H), 4.11-4.15 (m, 1H), 4.59 (m, 1H), 6.78-6.81 (m, 2H), 6.82-6.91 (m, 2H).

Step 3: Preparation of Compound L-1

To a solution of compound K-1 (220 mg) in methanol (20 mL) was added palladium (Pd-C; 10%, 50 mg) and stirred under a hydrogen balloon for 24 hours. The reaction was filtered through a pad of celite and concentrated and the residue was separated by silica gel column chromatography (ethyl acetate: methanol = 9: 1) to give compound L-1 (170 mg, 82%).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 1.25 (br s, 2H), 1.467 (s, 9H), 1.38-1.58 (m, 2H), 1.67-1.75 (m, 4H), 3.01-3.03 (m , 4H), 3.18-3.23 (m, 1H), 3.47 (m, 1H), 3.06-3.78 (m, 3H), 4.37 (m, 1H), 6.77-6.80 (m, 2H), 6.88-6.91 (m , 2H).

Step 4: Preparation of Compound M-1

Di (2-pyridyl) thiocarbonate, DPT; 133 mg, 0.56 mmol) in a solution of compound L-1 (170 mg, 0.47 mmol) in dichloromethane (10 mL). Add and stir for 1 hour. The reaction was concentrated and the residue was purified by silica gel column chromatography (EtOAc: Hexane == 1: 5) to give compound M-1 (20 mg, 52%).

Step 5: Preparation of Compound 119

Oxidation with 4-fluoro-1,2-phenylenediamine (Compound G-3 , 12 mg, 0.091 mmol) in a solution of isothiocyanate compound M-1 (31 mg, 0.076 mmol) in THF (10 mL) Add mercury (31 mg, 0.152 mmol) and stir at 60 ° C for 12 hours. The reaction mixture was filtered through a pad of celite, distilled water (10 mL) was added, extracted with EtOAc (10 mL), water was removed with sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane: ethyl acetate = 1: 1) to give compound 119 (20 mg, 52%).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 7.13-6.85 (m, 5H), 6.76 (t, J = 8.9 Hz, 1H), 5.70 (brs, 1H), 4.98-4.78 (m, 1H), 4.58 -4.41 (m, 1H), 3.85 (dd, J = 11.7, 5.2 Hz, 1H), 3.69-3.41 (m, 4H), 3.04 (d, J = 4.3 Hz, 4H), 1.77-1.66 (m, 4H ), 1.60-1.30 (m, 11 H).

[ Example  19] (3S, 4S) tert - butyl  3- (5- fluoro -1H- benzo Preparation of [d] imidazol-2-ylamino) -4- (4-morpholinophenoxy) pyrrolidine-1-carboxylate (Compound 120)

The same procedure as in Example 18, except that morpholine (Compound J-2 ) was used instead of piperidine (Compound J-1 ) from Compound 112 (48 mg, 0.10 mmol) obtained in Step 1 of Example 18. Compound 120 was obtained by the method.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 7.21-6.69 (m, 7H), 5.74 (brs, 1H), 5.03-4.77 (m, 1H), 4.60-4.41 (m, 1H), 3.92-3.70 ( m, 5H), 3.70-3.42 (m, 3H), 3.13-3.01 (m, 4H), 1.51 (s, 9H).

[ Example  20] (3S, 4S) tert - butyl  3- ( biphenyl -4- ylmethoxy ) -4- (5- fluoro Preparation of -1H-benzo [d] imidazol-2-ylamino) pyrrolidine-1-carboxylate (Compound 121)

Step 1: Preparation of Compound O-1

Sodium hydride (92 mg, 2.30 mmo) in a solution of azido alcohol compound B (500 mg, 2.19 mmol) obtained in step 1 of Example 1 in N, N-dimethylformamide (10 mL) was added at 0 ° C. Add and stir for 10 minutes. 4-Bromomethylbiphenyl (Compound N-1 , 541 mg, 2.19 mmol) is dissolved in anhydrous N, N-dimethylformamide (2 mL), added dropwise to the reaction, and stirred at room temperature for 5 hours. Ice water (20 mL) was added to the reaction, followed by extraction with ethyl acetate (2 x 20 mL), followed by removal of water with magnesium sulfate. After concentration under reduced pressure, the residue was separated by silica gel column chromatography (hexane: ethyl acetate: dichloromethane = 10: 1: 2) to give compound O-1 (357 mg, 41%).

¹ H NMR (300 MHz, CDCl ₃ ) δ 1.46 (s, 9H), 3.36-3.68 (m, 4H), 3.99-4.05 (m, 2H), 4.60 (m, 2H), 7.32-7.46 (m, 5H) , 7.59 (m, 4 H).

Step 2: Preparation of Compound P-1

Compound O-1 (357 mg, 0.90 mmol) was dissolved in anhydrous tetrahydrofuran (15 mL), triphenylphosphine (285 mg, 1.08 mmol) was added and stirred at room temperature for 4 hours. Distilled water (5 mL) was added to the reaction mixture, which was stirred for 24 hours at room temperature, extracted with ethyl acetate (2 x 15 mL), and water was removed with magnesium sulfate. After concentration under reduced pressure, the residue was separated by silica gel column chromatography (ethyl acetate) to give compound P- 1 (311 mg, 93%).

¹ H NMR (300 MHz, CDCl ₃ ) δ 1.46 (s, 9H), 3.14 (m, 1H), 3.36-3.50 (m, 2H), 3.60-3.75 (m, 3H), 4.52-4.65 (m, 2H) , 7.32-7.48 (m, 5 H), 7.58 (m, 4 H).

Step 3: Preparation of Compound Q-1

Compound Q-1 was obtained from compound P-1 (100 mg, 0.27 mmol) in the same manner as in Step 4 of Example 1 (111 mg, 100%).

¹ H NMR (300 MHz, CDCl ₃ ) δ 1.47 (s, 9H), 3.47-3.68 (m, 4H), 4.14 (br s, 2H), 4.63 (m, 2H), 7.33-7.47 (m, 5H), 7.57-7.60 (m, 4 H).

Step 4: Preparation of Compound 121

Compound Q-1 (111 mg, 0.27 mmol) was dissolved in anhydrous N, N-dimethylformamide (5 mL) and then 4-fluoro-1,2-phenylenediamine (Compound G-3 , 39 mg, 0.29 mmol) and stirred at 70 ° C. for 2 hours, then 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide (57 mg, 0.29 mmol) was added and stirred at 70 ° C. for 15 hours. Dilute the reaction with ethyl acetate (50 mL), wash with saturated sodium bicarbonate solution (50 mL), 1N aqueous hydrochloric acid solution (50 mL), and brine (20 mL), and then remove the moisture with magnesium sulfate. After concentration under reduced pressure, the residue was purified by silica gel column chromatography (hexane: ethyl acetate: dichloromethane = 1: 6: 2) to give compound 121 (97 mg, 71%).

¹ H NMR (300 MHz, CDCl ₃ ) δ 1.48 (s, 9H), 3.42 (m, 2H), 3.65-3.91 (m, 2H), 4.13-4.35 (m, 2H), 4.71 (m, 2H), 6.74 (m, 1H), 7.01 (m, 2H), 7.33-7.47 (m, 5H), 7.58 (m, 4H).

[ Example  21] tert - butyl  3- (5- fluoro -1H- benzo [d] imidazol-2- ylamino ) -4- (4-isopropylbenzyloxy) pyrrolidine-1-carboxylate (Compound 122)

From azido alcohol compound B obtained from step 1 of Example 1 Compound 122 was prepared in the same manner as in Example 20 using 4-isopropylbenzyl bromide (Compound N-2 ).

¹ H NMR (300 MHz, CDCl ₃ ) δ 1.26 (d, J = 7.1 Hz 6H), 1.48 (s, 9H), 2.86-3.00 (m, 1H), 3.30-3.46 (m, 1H), 3.63-3.87 (m, 2H), 4.07-4.31 (m, 3H), 4.51-4.71 (m, 2H), 6.60-6.86 (brs, 2H), 7.20-7.38 (m, 5H).

[ Example  22] 5- fluoro -N- (4- (4- isopropylbenzyloxy ) pyrrolidine -3- yl ) -1H-benzo [d] imidazol-2-amine (Compound 123)

Compound 122 (270 mg, 0.58 mmol) obtained in Example 21 was dissolved in trifluoroacetic acid: dichloromethane: water (70:30: 2.5 mixed solution, 10 mL) at 0 ° C., and then stirred at room temperature for 2 hours. The reaction was neutralized with saturated aqueous sodium hydrogen carbonate solution, extracted with ethyl acetate (50 mL), and water was removed with magnesium sulfate. After concentration under reduced pressure, the residue was purified by silica gel column chromatography (dichloromethane: methanol = 10: 1) to give compound 123 (200 mg, 94%).

¹ H NMR (300 MHz, CDCl ₃ ) δ 1.12 (d, J = 7.1 Hz, 6H), 2.84-2.96 (m, 1H), 3.10 (dd, J = 11.3, 4.4 Hz, 1H), 3.21-3.29 ( m, 1H), 3.49 (s, 1H), 3.50-3.62 (m, 2H), 4.19-4.29 (m, 2H), 4.60 (s, 2H), 6.78-6.89 (m, 1H), 6.93 (dd, J = 8.4, 2.1 Hz, 1H), 7.05-7.13 (m, 1H), 7.18-7.31 (m, 4H).

[ Example  23] 5- fluoro -N-((3S, 4S) -4- (4- isopropylbenzyloxy ) -1- (propylsulfonyl) pyrrolidin-3-yl) -1H-benzo [d] imidazol-2-amine (Compound 124)

Compound 123 (20.0 mg, 0.05 mmol) and diisopropylethylamine (9.50 mL, 50 mmol) obtained in Example 22 were dissolved in dichloromethane (10 mL) in propanesulfonyl chloride (0.1 M dichloro) at 0 ° C. Methane solution; 6.00 mL, 60 mmol), and then stirred at room temperature for 2 hours. The reaction was concentrated under reduced pressure and the residue was purified by silica gel column chromatography (hexane: ethyl acetate = 5: 1) to give compound 124 (20 mg, 84%).

¹ H NMR (300 MHz, CDCl ₃ ) δ 1.01 (t, J = 7.3 Hz, 3H), 1.23 (d, J = 7.0 Hz, 6H), 1.76-1.90 (m, 2H), 2.84-3.02 (m, 3H), 3.36 (dd, J = 2.1, 11.5 Hz, 1H), 3.55 (dd, J = 2.16, 10.1 Hz, 1H), 3.66-3.82 (m, 3H), 4.19-4.25 (m, 1H), 4.44 -4.52 (m, 1H), 4.53-4.69 (m, 2H), 6.69-6.79 (m, 1H), 6.94 (dd, J = 2.1, 8.6 Hz, 1H), 7.01-7.10 (m, 1H), 7.16 -7.29 (m, 4 H).

[ Example  24] 5- fluoro -N-((3S, 4S) -4- (4- isopropylbenzyloxy ) -1- (phenylsulfonyl) pyrrolidin-3-yl) -1H-benzo [d] imidazol-2-amine (Compound 125)

Compound 125 was obtained in the same manner as in Example 23, except that phenylpropanesulfonyl chloride was used instead of propanesulfonyl chloride from compound 123 (20.0 mg, 0.05 mmol) obtained in Example 22 (20 mg, 79%). ).

¹ H NMR (300 MHz, CDCl ₃ ) δ 1.23 (d, J = 6.8 Hz, 6H), 2.83-2.95 (m, 1H), 3.11-3.21 (m, 1H), 3.42-3.54 (m, 2H), 3.65-3.75 (m, 1H), 4.05-4.12 (m, 1H), 4.27-4.36 (m, 1H), 4.37-4.48 (m, 2H), 6.73 (t, J = 9.0 Hz, 1H), 6.86- 6.97 (m, 1H), 6.97-7.07 (m, 3H), 7.14 (d, J = 7.8 Hz, 2H), 7.26 (s, 2H), 7.51 (d, J = 7.3 Hz, 2H), 7.60 (t , J = 7.2 hz. 1H), 7.80 (d, J = 7.4 Hz, 2H).

[ Example  25] 2-((3S, 4S) -3- (5- fluoro -1H- benzo [d] imidazol-2- ylamino ) -4- (4- isopropyl benzyloxy ) pyrrolidine Preparation of -1-yl) ethanol (Compound 126)

Compound 123 (20.0 mg, 0.05 mmol) obtained in Example 22 was dissolved in N, N-dimethylformamide (2 mL), followed by sodium carbonate (9 mg, 0.10 mmol) and 2-iodineethanol (4 mL, 0.05 mmol). ) And stir at 50 ° C. for 16 hours. Distilled water (5 mL) was added to the reaction, followed by extraction with ethyl acetate (10 mL), followed by removal of water with sodium sulfate. After concentration under reduced pressure, the reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (dichloromethane: methanol = 10: 1) to give compound 126 (14 mg, 63%).

¹ H NMR (300 MHz, CDCl ₃ ) δ 1.23 (d, J = 7.0 Hz, 6H), 2.50-2.60 (m, 1H), 2.66-2.99 (m, 5H), 3.76 (t, J = 4.8 Hz, 2H), 3.97-4.06 (m, 1H), 4.09-4.19 (m, 1H), 4.44-4.59 (m, 2H), 6.66-6.77 (m, 1H), 6.92 (dd, J = 2.2, 9.3 Hz, 1H), 6.97-7.05 (m, 1H), 7.17-7.29 (m, 4H).

[ Example  26] 2-((3S, 4S) -3- (5- fluoro -1H- benzo [d] imidazol-2- ylamino ) -4- (4- isopropyl benzyloxy ) pyrrolidine Preparation of -1-yl) acetamide (Compound 127)

Compound 123 (20.0 mg, 0.05 mmol) obtained in Example 22 was dissolved in N, N-dimethylformamide (2 mL), followed by potassium carbonate (9 mg, 60 mmol) and 2-chloroacetamide (5 mg, 50 mmol) is added and stirred at room temperature for 16 hours. Distilled water (5 mL) was added to the reaction, followed by extraction with ethyl acetate (10 mL), followed by removal of water with sodium sulfate. After concentration under reduced pressure, the reaction product was concentrated under reduced pressure, and then the residue was purified by silica gel column chromatography (dichloromethane: methanol = 10: 1) to give compound 127 (12 mg, 52%).

¹ H NMR (300 MHz, CDCl ₃ ) δ 1.25 (d, J = 6.7 hz, 6H), 2.77-2.98 (m, 3H), 3.08-3.33 (m, 4H), 4.00-4.05 (m, 2H), 4.45 (d, J = 10.8 hz, 1H), 4.60 (d, J = 10.8 hz, 1H), 5.94-6.07 (brs, 1H), 6.60-6.68 (brs, 1H), 6.68-6.78 (m, 1H) , 6.85-7.03 (m, 2 H), 7.21-7.31 (m, 3 H).

On the other hand, the piperidine compound represented by the formula (I) according to the present invention can be formulated in various forms according to the purpose.

[ Test Example  One] BACE  Inhibitory efficacy test

In order to investigate the effect of inhibiting beta-secretase activity by 4- (benzimidazol-2-ylamino) pyrrolidine derivative of the present invention, the following experiment was performed.

To the BACE1 substrate solution (Rh-EVNLDAEFK-Quencher, 750 nM in assay buffer), 10 μl of the desired concentration of test drug is added and mixed lightly, followed by baculovirus-expressed BACE1 enzyme (Bacculovirus-expressed) 10 μl of a 1.0 unit / mL in assay buffer solution was added to induce a reaction, and then incubated at room temperature for 60 minutes. Immediately after completion of the culture, 10 μl of a stop solution was added to the reaction solution to stop the reaction, and the fluorescence was measured at 545 nm (excitation) and 585 nm (emission) using a FlexStation to measure enzyme activity. The total volume of the above experiment was 40 μl and was performed on a 384-black microwell plate. Detailed compositions of enzymes, substrates, stop solutions, assay buffers, etc. used in the test are shown in Table 1 below.

Description Composition BACE1 Enzyme
(BACE1 enzyme) 1 U / mL in 50 mM Tris (ph 7.5), 10% glycerol BACE1 Substrate
(BACE1 substrate) Rh-EVNLDAEFK-Quencher (750 nM) BACE1 Termination Solution
(BACE1 stop solution) 2.5 M sodium acetate BACE1 Assay Buffer
(BACE1 assay buffer) 50 mM sodium acetate (pH 4.5)

The test results are shown in Table 2 below, and 4- (benzimidazol-2-ylamino) pyrrolidine derivatives according to the present invention are judged to have an excellent effect of inhibiting BACE activity.

Compound number BACE Inhibition Assay (IC ₅₀ = μM) Compound number BACE Inhibition Assay (IC ₅₀ = μM) Compound 101 0.21 Compound 114 0.67 Compound 106 0.45 Compound 115 0.72 Compound 107 0.05 Compound 116 3.15 Compound 108 0.44 Compound 119 0.70 Compound 109 0.19 Compound 120 3.15 Compound 110 0.36 Compound 121 0.12 Compound 112 0.07 Compound 122 0.73 Compound 113 1.35 Compound 123 > 5.0 Compound 118 4.44 Compound 124 0.64 Compound 102 0.66 Compound 125 0.48 Compound 103 0.72 Compound 126 > 5.0 Compound 104 0.39 Compound 127 > 5.0 Compound 105 0.57

[ Test Example  2] PC12 PC12tet - off pTRE2puro - APPsw stable   Used BACE  Inhibitory efficacy test

In order to determine the effect of inhibiting beta-secretase activity by 4- (benzimidazol-2-ylamino) pyrrolidine derivatives of the present invention, the following experiments based on cells were performed by Invitrogen, Inc. Using an ELISA assay kit (KHB3482) Respectively.

The PC12tet-off pTRE2puro-APPsw stable cell line was prepared in RPMI 1640 (10% FBS, 5% HS, 1% Penicillin / Streptomycin) at 100 µg / ml G418 (Clontech), 1 µg / ml Doxicyclin (Clontech), 3µg / ml Puromycin (Clontech) was added and cultured at 5% CO ₂ , 37 ° C. The cultured cells were divided into 4 × 10 ⁴ cells / well in a 0.05% PEI-coated 6-well plate and after 24 hours, exchanged with 2 ml of condition media (5% HS, 1% FBS, 0.5% P / S) for 24 hours. It was. After treating the compound of Table 2 at 1 μM concentration and reacting for 24 hours, 50 μl of the supernatant was transferred to a microwell plate, 50 μl of beta amyloid antibody (anti-hu-Ab40) was added thereto, and then washed after 3 hours of incubation. The antibody was added again and incubated for 30 minutes. Chromogen and stop solution were added to the plate reacted with the antibody to stop the reaction, and then the absorbance was measured at 450 nm using FlexStation to measure the amount of beta amyloid.

As a result, it was shown that the compounds of the compounds 106, 112 and 102 of the present invention have an effect of inhibiting BACE activity of 50% or more at a concentration of 1 mM or less.

Claims (7)

4- (benzimidazol-2-ylamino) pyrrolidine derivative represented by Formula 1 or a pharmaceutically acceptable salt thereof:
[Chemical Formula 1]

In Formula 1,
A is CH or N;
Y is a bond, -CH ₂ -or -C (= 0)-;
R ₁ is halogen, (C1-C10) alkyl, (C1-C10) alkoxy, (C6-C12) aryl, (C3-C12) heteroaryl, (C6-C12) aryloxy, NR ₁₁ R ₁₂ , 5-membered to 7-membered cyclicamino or 5- to 7-membered heterocyclic amino comprising at least one heteroatom selected from nitrogen, oxygen and sulfur;
R ₁₁ and R ₁₂ are each independently (C 1 -C 10) alkyl or (C 6 -C 12) aryl;
R ₂ is (C 1 -C 10) alkyl, (C 1 -C 10) alkoxy or halogen;
R ₃ is hydrogen, (C 1 -C 10) alkyl,-(CH ₂ ) _m -COR ₄ ,-(CH ₂ ) _m -COOR ₄ ,-(CH ₂ ) _m -CONR ₄ R ₅ or -SO ₂ R ₄ ;
R ₄ and R ₅ are each independently hydrogen, (C 1 -C 10) alkyl or (C 6 -C 12) aryl;
m is an integer from 0 to 5;
Alkyl of said R _1, alkoxy, aryl or aryloxy, the alkyl or alkoxy group of R _2, the R ₃ alkyl and R _4, R _5, alkyl or aryl of R ₁₁ and R ₁₂ are halogen, alkyl, aryl, and It may be further substituted with one or more substituents selected from the group consisting of hydroxy.
The method of claim 1,
R ₁ is (C ₁ -C 10) alkyl, (C 6 -C 12) aryl, halogen substituted (C 6 -C 12) aryl, (C 6 -C 12) aryloxy, di (C 1 -C 10) alkylamino, 5-membered to 7 Cyclic amino or 5- to 7-membered heterocyclic amino; R ₂ is (C 1 -C 10) alkyl, (C 1 -C 10) alkoxy or halogen; R ₃ is hydrogen, hydroxy (C 1 -C 10) alkyl, —COR ₄ , —CH ₂ COOR ₄ , —COOR ₄ , —CH ₂ CONR ₄ R ₅ or —SO ₂ R ₄ ; R ₄ and R ₅ are each independently hydrogen, (C 1 -C 10) alkyl or (C 6 -C 12) aryl; 4- (benzimidazol-2-ylamino) pyrrolidine derivative or a pharmaceutically acceptable salt thereof.
3. The method of claim 2,
R ₁ is methyl, n-propyl, i-propyl, phenyl, phenyl substituted phenyl, phenoxy, dimethylamino, piperidino or morpholino; R ₂ is methyl, methoxy, chloro or fluorine; R ₃ is hydrogen, hydroxyethyl,

,

,

,

,

,

or

Phosphorus 4- (benzimidazol-2-ylamino) pyrrolidine derivative or a pharmaceutically acceptable salt thereof.
The method of claim 1,
The 4- (benzimidazol-2-ylamino) pyrrolidine derivative is any one selected from compounds of the following structure 4- (benzimidazol-2-ylamino) pyrrolidine derivative or Pharmaceutically acceptable salts.

Beta-secretase containing 4- (benzimidazol-2-ylamino) pyrrolidine derivative according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof as an active ingredient. Activity inhibitor composition of (BACE).
Prevention of neurodegenerative diseases containing 4- (benzimidazol-2-ylamino) pyrrolidine derivatives according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof as an active ingredient And therapeutic pharmaceutical compositions.
The method according to claim 6,
The neurodegenerative disease is Alzheimer's or Down syndrome.