PL241191B1 - Derivatives of 6-(2-chloro-[1,1'-biphenyl]-3-ylo)benzo-1,4-dioxne for use in immune therapy of tumours - Google Patents

Abstract

The subject of the application is a compound of general formula (I): where R1 is -H, -OCH2CN, an alkoxy group consisting of 1 to 6 carbon atoms, preferably containing alkane and methylcycloalkane fragments, an N, N-dialkyl group, a diaryl group or an alkylaryl carbamate, R2 is a substituent selected from the structures represented by general formula (I) b, or a pharmaceutically acceptable salt thereof. The application also includes a pharmaceutical composition containing a pharmaceutically effective dose of the subject compound.

Description

Description of the invention

The subject of the invention are new 6- (2-chloro- [1,1'-biphenyl] -3-yl) benzo-1,4-dioxane derivatives which are small molecule inhibitors targeting the PD-1 / PD-L1 immune response checkpoint that can be found application in cancer immunotherapy.

Immunotherapy based on inhibitors of checkpoints, such as PD-1, CTLA-4, etc. results in sustained therapeutic responses and longer survival in patients with various types of cancer ¹ ' ³ . Advances in this area led to the journal Science announcing the 'Breakthrough of the Year' immunotherapy in 2013, while in 2018 James P. Allison from the US and Tasuku Honjo from Japan were awarded the Nobel Prize in Medicine and Physiology for developing methods to unblock the immune system.

One of the ways in which tumor cells evade immune surveillance by allowing tumors to grow is through complex immunosuppressive mechanisms, which usually play a role in controlling normal antibacterial and antiviral immune responses and prevent excessive immune response and associated immunopathologies. For example, strongly or chronically activated T cells upregulate PD-1 expression such that cells will express PD-L1 upon exposure to T cell-derived IFN? CD8 + T cells via PD-L1 / PD-1.

PD-1 or programmed death receptor 1 (CD279) is a glycoprotein protein with a mass of about 55 kDa consisting of 288 amino acids from the B7-CD28 protein family. It consists of the extracellular IgV-like domain, transmembrane and cytoplasmic domains. PD-1 acts as a negative regulator of the immune response. It is encoded by the pdcdl gene located on the second chromosome (2q37) and is expressed on the surface of activated T and B lymphocytes, responsible for the regulation of immune tolerance.

The PD-1 ligand (PD-L1, CD270 or B7-H1) is a 33 kDa glycoprotein protein with the gene located on chromosome 9p24. This type I transmembrane protein consists of 290 amino acids and has IgV and IgC domains in the extracellular part. Its overexpression has been observed in many cells of the immune system such as macrophages, B lymphocytes and many types of non-hematopoietic cells ⁴ . The interaction between PD-1 and its PD-L1 ligand results in the suppression of, inter alia, T cell proliferation, cytokine secretion, and cytotoxic function of lymphocytes. This pathway is also often used by cancer cells as well as viruses and bacteria to weaken the body's natural immune response and facilitate spread throughout the affected organism ⁵ . Blockade of this pathway, e.g. by specific antibodies blocking the action of PD1- / PD-L1, restores cytotoxic functions of T lymphocytes, normalizing the immune response of the organism and is one of the most important elements of cancer immunotherapy.

At present, all registered or clinical-phase compounds that directly block the interaction of the PD-1 protein with its ligand are based on monoclonal antibodies or their combinations with each other or with chemotherapy. The only small molecule compound currently in Phase I clinical trials in the United States that was expected to directly interact with PD1 / PD-L1 proteins (CA-170) has recently been characterized as interacting with the PD-1 / PD-L1 ⁶ pathway. However, small molecule inhibitors of the PD-1 / PD-L1 interaction are an interesting alternative to antibody-based therapies due to their much lower production costs, the ability to be administered orally instead of intravenously, better pharmacokinetics and potentially no adverse effects related to the patient's immunity so characteristic of therapies based on antibodies ⁷ .

The aim of the invention is to provide new compounds that could be used as an inhibitor of the PD-1 / PD-L1 interaction.

The invention relates to compounds and their uses as defined in the appended claims.

In particular, the invention relates to a PD-1 / PD-L1 interaction inhibitor of general formula (I) (Scheme 1),

Scheme 1. Structure of inhibitors based on 6- (2-chloro- [1,1'-biphenyl] -3-yl) benzo-1,4-dioxane core (I)

PL241 191 Β1 where Ri is -H, -OCH2CN, an alkoxy group consisting of 1 to 6 carbon atoms (containing alkane and methylcycloalkane fragments), Ν, Ν-dialkyl, diaryl or alkylarylcarbamate, R2 are substituents that can be represented by with the help of the following structures:

or a pharmaceutically acceptable salt thereof that finds use as a medicament in the treatment of neoplastic diseases.

These inhibitors dimerize the PD-L1 protein, preventing its interaction with the PD-1 protein, as confirmed by NMR. The activity of the compounds in dissociating the PD-1 / PD-L1 complex has been confirmed by Homogenous Time Resolved Fluorescence (HTRF) tests.

For a better understanding of the essence of the invention, the present description has been enriched with detailed examples of its implementation, which are presented below.

^1H and ¹³ CNMR spectra were recorded on a Bruker Avance 600 MHz spectrometer expressing chemical shifts (δ) in ppm and coupling constants (J) in Hz. TMS was used as a template and internal standard. In addition, chemical shifts were analyzed with respect to the solvent peaks (DMSO-de, CDCl 3). The compounds were purified by column chromatography using a Grace Reveleris X2 flash chromatograph. The progress of the reaction was monitored by visualization of Merck 60 254 to 365 nm silica gel TLC plates.

Example 1. (3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro- [1, T-biphenyl] -4-yl) methanol

Intermediate product 1.1. (2 ^, -amino-3 '- (benzo-1,4-dioxan-6-yl) - [1, r-biphenyl] -4-yl) methanol

And OH

M (dppi) Cl, * DCM K ₂ CO ₃ dioxan / water (2/1, v, v)

(1.1)

A three-necked round bottom flask was charged with 2,6-dibromoaniline (5.02 g, 20.00 mmol, 1.2 eq.), 1,4-benzodioxin-6-boronic acid (3.00 g, 16.67 mmol, 1 , 0 eq.), K2CO3 (9.22 g, 66.68 mmol, 4.0 eq.), And a dioxane / water mixture (30 ml, 2/1, v, v). The solution thus obtained was deoxygenated for 30 minutes by purging with argon, then Pd (dppf) Cl2-DCM (0.68 g, 0.83 mmol, 0.05 eq.) Was added.

PL241 191 Β1

The contents of the flask were heated at 80 ° C for 3 hours under an inert atmosphere, then 4- (hydroxymethyl) phenylboronic acid (3.04 g, 20.00 mmol, 1.2 eq.) Was added and heated to the same temperature. for the next 3 hours. At the end of this time, the contents of the flask were transferred to a separating funnel and extracted three times with ethyl acetate (15 ml each). The organic phases were combined, dried over anhydrous MgSO4 and purified by flash chromatography (S1O2, hexane / ethyl acetate, 4/1). The product 1.1 was obtained in a yield of 32% (1.78 g).

¹ H NMR (600 MHz, CDCl 3) δ [ppm]: 7.53 (m, 2H), 7.47 (d, J = 8.2 Hz, 2H), 7.12 (m, 2H), 7, 05 (d, J = 2.00 Hz, 1H), 7.00 (dd, J = 8.3, 2.0 Hz, 1H), 6.97 (d, J = 8.2 Hz, 1H), 6.88 (t, J = 7.5Hz, 1H), 4.75 (S, 2H), 4.31 (S, 4H), 3.90 (Wide, 2H), 2.23 (Wide, 1H ) ¹³ C NMR (151 MHz, CDCl 3)? [Ppm]: 143.7; 142.9; 141.0; 139.9; 139.2; 133.0; 129.8; 129.6; 129.5; 127.5; 127.5; 127.5; 122.4; 118.1; 117.6; 65.1; 64.5; 64.4

End product 1. (3 ^, - (benzo-1,4-dioxan-6-yl) -2 ^, -chloro [1, r-biphenyl] -4-yl) methanol

NaNO ₂ CoCl

HCl dioxane aqueous (1.1)

(1)

In a three-neck round bottom flask equipped with a thermometer was placed (2'-amino- [1,1 ': 3', 1'-terphenyl] -4-yl) methanol (0.22 g, 0.63 mmol, 1.0 eq. ) and dissolved in 2 ml of dioxane, then 1 ml of 15% HCl was added. The resulting mixture was cooled in an ice bath, then a solution of NaNO2 (0.10 g, 1.51 mmol, 2.4 eq.) In 2 mL of water was added dropwise to the flask, keeping the temperature of the reaction mixture below 5 ° C and stirred for 5 minutes. After this time, a solution of CuCl (0.12 g, 1.26 mmol, 2 eq.) In 0.4 ml of concentrated HCl was added dropwise to the mixture keeping the temperature below 5 ° C. After adding the reagent, the mixture was warmed to room temperature and stirred for 40 minutes. At the end of this time, the contents of the flask were transferred to a separating funnel, 5 ml of water were added and the mixture was extracted three times with ethyl acetate (10 ml each). The organic phases were combined, dried over anhydrous MgSO 4 and purified by flash chromatography (S1O2, hexane / ethyl acetate, 4/1). The product 1 was obtained in a yield of 53% (0.12 g).

¹ H NMR (600 MHz, CDCl 3) δ [ppm]: 7.48 (m, 4H), 7.36-7.30 (m, 3H), 7.03 (d, J = 1.9 Hz, 1H ), 6.98-6.95 (m, 2H), 4.79 (s, 2H), 4.33 (s, 4H), 1.75 (s, 1H) ¹³ CNMR (151 MHz, CDCh) δ [ppm] 143.2; 143.0; 141.3; 141.1; 140.1; 139.6; 133.4; 131.0; 130.5; 130.2; 129.8; 126.7; 126.4; 122.8; 118.6; 116.9; 65.2; 64.5; 64.4

Example 2. (3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1,1-biphenyl] -4-yl) methanol

Intermediate product 2.1. (2-methoxy-4- (4,4,5,5-tetramethyl-1,3,2-dioxyborolan-2-yl) phenyl) methanol.

anhydrous dioxane

Pd (dppf) Cl ₂ * DCM AcOK

In a two-necked round bottom flask was placed bis (pinacolato) diborane (1.43 g, 5.53 mmol, 1.2 eq), anhydrous potassium acetate (1.38 g, 13.82 mmol, 3.0 eq), alcohol 4 -bromo-2-methoxybenzyl (1.02 g, 4.61 mmol, 1.0 eq) and anhydrous dioxane (20 mL). The solution thus obtained was deoxygenated for 30 minutes by purging with argon, then Pd (dppf) ChDCM (0.19 g, 0.02 mmol, 0.05 eq,) was added. The contents of the flask were heated at 80 ° C for 3 hours. After this time, 10 ml of water was added to the flask, the mixture was transferred to a separating funnel and extracted three times with ethyl acetate (15 ml each). The organic phases were combined, dried over anhydrous MgSO4 and purified by flash chromatography (SiO2, hexane / ethyl acetate, 4/1). The product 2.1 was obtained in a yield of 75% (0.93 g).

¹ H NMR (600 MHz, CDCl 3) δ [ppm]: 7.42 (d, J = 7.3 Hz, 1H), 7.29 (m, 2H), 4.70 (s, 2H), 3, 91 (s, 3H), 2.40 (Wide, 1H), 1.35 (s, 12H) ¹³ CNMR (151 MHz, CDCl 2) δ [ppm]: 156.9; 132.2; 128.0; 127.6; 115.6; 83.9; 62.2; 55.4; 24.9

PL241 191 Β1

Intermediate product 2.2. 6- (3-bromo-2-chlorophenyl) benzo-1,4-dioxane

Fd (dppf) Ck * DCM KjCOj dioxane, water

(ί.2)

In a three-necked round bottom flask was placed 1,3-dibromo-2-chlorbenzene (5.16 g, 20.09 mmol, 1.1 eq.), 1,4-benzodioxin-6-boronic acid (3.12 g, 167, 40 mmol, 1.0 eq.), K2CO3 (9.62 g, 69.60 mmol, 4.0 eq.) And a dioxane / water mixture (120 ml, 2/1, v, v). The solution thus obtained was deoxygenated for 30 minutes by purging with argon, then Pd (dppf) Cl2-DCM (0.71 g, 0.87 mmol, 0.05 eq.) Was added. The contents of the flask were heated at 80 ° C for 5 hours under an inert gas atmosphere. At the end of this time, the contents of the flask were transferred to a separating funnel and extracted three times with ethyl acetate (40 ml each). The organic phases were combined, dried over anhydrous MgSO4 and purified by flash chromatography (SiO2, hexane / ethyl acetate, 4/1). The product 2.2 was obtained in a yield of 47% (2.65 g).

¹ H NMR (600 MHz, CDCl 3) δ [ppm]: 7.59 (dd, J = 8.0; 1.6 Hz, 1H), 7.24 (dd, J = 7.6; 1.6 Hz , 1H), 7.13 (t, 4 = 7.8 Hz, 1H), 6.93-6.91 (m, 2H), 6.87 (dd, J = 8.3, 2.1 Hz, 1H), 4.30 (s, 4H) ¹³ C NMR (151 MHz, CDCl 3) δ [ppm]: 143.5; 143.1; 142.3; 132.9; 132.9; 132.6; 130.2; 127.4; 123.8; 122.5; 118.3; 117.0; 64.5; 64.4

End product 2. (2'-chloro-3 '- (benzo-1,4-dioxan-6-yl) -3-methoxy- [1, r-biphenyl] -4-yl) methanol

(2.1)

(2.2)

Pd (dppf) _C12 * DCM

K ₂ CO ₃ dioc & an.-water

6- (3-Bromo-2-chlorophenyl) benzo-1,4-dioxane (0.46 g, 1.41 mmol, 1.0 eq.), (3 '- (2.3- dihydrobenzo [b] [1,4] dioxin-6-yl) -3-methoxy-2'-methyl- [1,1'-biphenyl] -4-yl) methanol (0.37 g, 1.41 mmol, 1.0 eq.), K2CO3 (0.78 g, 5.63 mmol, 4.0 eq.), And a dioxane / water mixture (30 ml, 2/1, v, v). The solution thus obtained was deoxygenated for 30 minutes by purging with argon, then Pd (dppf) Cl2-DCM (0.06 g, 0.07 mmol, 0.05 eq.) Was added. The contents of the flask were heated at 80 ° C for 5 hours under an inert gas atmosphere. At the end of this time, the contents of the flask were transferred to a separating funnel and extracted three times with ethyl acetate (40 ml each). The organic phases were combined, dried over anhydrous MgSO4 and purified by flash chromatography (SiO2, hexane / ethyl acetate, 1/1). The product 2 was obtained in a yield of 70% (0.38 g).

¹ H NMR (600 MHz, CDCl 3) δ [ppm]: 7.35 (d, J = 7.4 Hz, 1H), 7.33-7.28 (m, 3H), 7.02 (dd, J = 7.5; 1.5 Hz, 1H), 7.00 (m, 2H), 6.99-6.92 (m, 2H), 4.74 (d, J = 6.4 Hz, 2H) , 4.31 (s, 4H), 3.89 (s, 3H), 2.30 (t, 4 = 6.5 Hz, 1H) ¹³ C NMR (151 MHz, CDCl 3) δ [ppm]: 156. 9; 143.2; 143.0; 141.4; 141.1; 141.1; 133.3; 131.0; 130.6; 130.1; 128.4; 128.3; 126.4; 122.8; 121.8; 118.6; 116.9; 111.9; 64.5; 64.4; 62.0; 55.4;

Example 3. (3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-ethoxy- [1,1'-biphenyl] -4-yl) methanol

Intermediate product 3.1. 4-bromo-2-hydroxybenzyl alcohol

L1A1H4

EtjO anhydrous

(3.1)

LiAIH4 (2.62 g, 69.12 mmol, 3 eq.) Was added to a two-neck round bottom flask equipped with a reflux condenser and dropping funnel under an inert gas (Ar) atmosphere and anhydrous Et2O (40 mL) was added. A solution of 4-bromosalicylic acid (5.00 g, 23.04 mmol, 1 eq.) In anhydrous Et 2 O (100 ml) was added dropwise to the vigorously stirred suspension then left on the stirrer overnight. The next day the mixture was cooled in an ice bath and ethyl acetate was added in small portions until gas evolution ceased, then saturated NH 4 Cl solution was added in small portions until gas evolution ceased. After neutralization of the reducing agent, the solution was acidified with 2M HCl to pH ~ 4 and extracted five times with Et2O (70 ml each). The organic layers were combined, dried over anhydrous MgSO4

PL241 191 Β1 and the solvent was evaporated. The product was purified by flash column chromatography (hexane / ethyl acetate, 1/1, v / v) to give compound 3.1 in 89% yield (4.16 g).

^1H NMR (600 MHz, DMSO-d6) δ [ppm]: 9.85 (s, 1H), 7.24 (d 'J = 8.1 Hz, 1H), 6.96 (dd, J = 8 0;

1.9 Hz, 1H), 6.94 (d, J = 1.9 Hz, 1H), 4.44 (s, 2H).

¹³ C NMR (151 MHz, DMSO-d ₆ ) δ [ppm]: 155.7; 129.4; 128.8; 121.9; 119.8; 117.5; 58.2

Intermediate product 3.2. 4-bromo-2-ethoxybenzyl alcohol

(3.1)

CS2CO ₃

DMF anhydrous

(3J)

4-bromo-2-hydroxybenzyl alcohol (0.50 g, 2.46 mmol, 1.0 eq.), Bromoethane (0.82 ml, 2.71 mmol, 1.1 eq.), Cs2CO3 were placed in a round bottom flask. (1.20 g, 3.69 mmol, 1.5 eq.) And anhydrous DMF (8 mL). The resulting mixture was left on a magnetic stirrer overnight, then the reaction progress was checked by TLC (S1O2, hexane / ethyl acetate, 2/1, v / v). After complete conversion of the alcohol, the mixture was transferred to a separating funnel, added with 10 ml of water and extracted three times with ethyl acetate (15 ml each). The organic layers were combined, dried over anhydrous MgSO4, and the solvent was evaporated. The product was purified by flash column chromatography (hexane / ethyl acetate, 2/1, v / v) to give compound 3.2 in 91% yield (0.52 g).

¹ H NMR (600 MHz, CDCl 3) δ [ppm]: 7.07 (d, J = 7.9 Hz, 1H), 6.99 (dd, J = 8.0; 1.7 Hz, 1H), 6.92 (d, J = 1.6 Hz, 1H), 4.56 (s, 2H), 4.00 (q, J = 7.0 Hz, 2H), 2.13 (Wide, 1H), 1.37 (t, J = 7.0 Hz, 3H) ¹³ CNMR (151 MHz, CDCl 3) δ [ppm]: 157.3; 129.7; 128.3; 123.5; 121.9; 114.7; 64.0; 61.6; 14.8 Intermediate product 3.3. (2-ethoxy-4- (4,4,5,5-tetramethyl-1,3,2-dioxyborolan-2-yl) phenyl) methanol

Pil (dppDa ₂ * DCM AcOK.

anhydrous dioxane

0Ϊ)

(3.3)

In a two-necked round bottom flask was placed bis (pinacolato) diborane (0.55 g, 2.17 mmol, 1.2 eq), anhydrous potassium acetate (0.53 g, 5.43 mmol, 3.0 eq), alcohol 4 -bromo-2-ethoxybenzyl (0.42 g, 1.81 mmol, 1.0 eq,) and anhydrous dioxane (10 mL). The solution thus obtained was deoxygenated for 30 minutes by purging with argon, then Pd (dppf) Cl2-DCM (0.07 g, 0.09 mmol, 0.05 eq,) was added. The contents of the flask were heated at 80 ° C for 3 hours. After this time, 10 ml of water was added to the flask, the mixture was transferred to a separating funnel and extracted three times with ethyl acetate (10 ml each). The organic phases were combined, dried over anhydrous MgSO4 and purified by flash chromatography (S1O2, hexane / ethyl acetate, 4/1). The product 3.3 was obtained in a yield of 81% (0.41 g).

¹ H NMR (600 MHz, CDCl 3) δ [ppm]: 7.39 (m, 1H), 7.28 (m, 2H), 4.70 (s, 2H), 4, W (q, J = 7 , 0 Hz, 2H), 2.44 (Wide, 1H), 1.44 (t, J = 7.0 Hz, 3H), 1.34 (s, 12H) ¹³ CNMR (151 MHz, CDCh) δ [ ppm]: 156.4; 132.3; 128.0; 127.5; 116.6; 83.9; 63.6; 62.6; 24.9; 15.0

End product 3. (3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-ethoxy- [1, r-biphenyl] -4-yl) methanol

(3)

In a three-necked round bottom flask was placed 6- (3-bromo-2-chlorophenyl) benzo-1,4-dioxane (0.50 g, 1.53 mmol, 1.2 eq.), (2-ethoxy-4- (4 , 4,5,5-tetramethyl-1,3,2-dioxyborolan-2-yl) phenyl) methanol (0.35 g, 1.27 mmol, 1.0 eq.), K2CO3 (0.70 g, 5 , 10 mmol, 4.0 eq.), And a dioxane / water mixture (15 mL, 2/1, v, v). The solution thus obtained was deoxygenated for 30 minutes by purging with argon, then Pd (dppf) Cl2-DCM (0.05 g, 0.06 mmol, 0.05 eq.) Was added. The contents of the flask were heated at 80 ° C for 5 hours under an inert gas atmosphere. At the end of this time, the contents of the flask were transferred to a separating funnel and extracted three times with ethyl acetate (20 ml each). The organic phases were combined, dried over anhydrous MgSO4 and purified by flash chromatography (SiO2, hexane / ethyl acetate, 1/1). The product 3 was obtained in a yield of 38% (0.19 g).

PL241 191 Β1 ¹ NMR (600 MHz, CDCl ₃ ) δ [ppm]: 7.38 (m, 1 Η), 7.35-7.31 (m, 3Η), 7.05-7.03 (m , 2Η), 7.02 (m, 1H), 6.99-6.95 (m, 2H), 4.79 (s, 2H), 4.33 (s, 4H), 4.16 (q, J = 7.0 Hz, 2H), 2.58 (Sbroad, 1H), 1.48 (t, J = 7.0 Hz, 3H) ^13C NMR (151 MHz, CDCl3) δ [ppm]: 156. 3; 143.2; 143.0; 141.4; 141.1; 140.9; 133.3; 131.0; 130.5; 130.1; 128.4; 128.4; 126.4; 122.8; 121.7; 118.6; 116.9; 112.8; 64.5; 64.4; 63.7; 62.1; 14.9

Example 4. 2 - ((3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro -) - 4- (hydroxymethyl) - [1, r-biphenyl] -3-yl) oxy ) acetonitrile

Intermediate product 4.1. 2- (5-bromo-2- (hydroxymethyl) phenoxyacetamide

C ^ COj ⁺ Br ^^ CN

DMF anhydrous (3.1)

(4.1)

4-bromo-2-hydroxybenzyl alcohol (0.50 g, 2.46 mmol, 1.0 eq.), Bromoacetonitrile (0.33 ml, 4.92 mmol, 2.0 eq.), Cs2CO3 were placed in a round bottom flask. (0.68 g, 4.92 mmol, 2.0 eq.) And anhydrous DMF (10 mL). The resulting mixture was left on a magnetic stirrer overnight and then the reaction progress was checked by TLC (S1O2, hexane / ethyl acetate, 4/1, v / v). After complete conversion of the alcohol, the mixture was transferred to a separating funnel, added with 10 ml of water and extracted three times with ethyl acetate (15 ml each). The organic layers were combined, dried over anhydrous MgSO4, and the solvent was evaporated. The product was purified by flash column chromatography (hexane / ethyl acetate, 4/1, v / v) to give compound 4.1 in 43% yield (0.25 g).

¹ H NMR (600 MHz, CDCl 3) δ [ppm]: 7.31 (d, J = 8.1 Hz, 1H), 7.25 (dd, J = 8.1, 1.6 Hz, 1H), 7.08 (d, J = 1.7 Hz, 1H), 4.82 (s, 2H), 4.67 (s, 2H) ¹³ C NMR (151 MHz, CDCl 3) δ [ppm]: 154.6; 130.5; 129.5; 126.5; 122.0; 115.5; 114.6; 60.3; 54.1

Intermediate product 4.2. (2- (2-hydroxymethyl) -5- (4,4,5,5-tetramethyl-1,3,2-dioxyborolan-2-yl) phenoxy) acetonitrile

PtKdppOChTłCM AcOK.

anhydrous dioxane (4.1)

(4.2)

In a two-necked round bottom flask was placed bis (pinacolato) diborane (0.57 g, 2.24 mmol, 1.1 eq), anhydrous potassium acetate (0.60 g, 6.12 mmol, 3.0 eq.), 2- (5-bromo-2- (hydroxymethyl) phenoxyacetamide (0.49 g, 2.04 mmol, 1.0 eq,) and anhydrous dioxane (10 ml) The solution thus obtained was deoxygenated for 30 minutes by flushing with argon, then added Pd (dppf) ChDCM (0.05 g, 0.06 mmol, 0.03 eq.). The contents of the flask were heated at 80 ° C for 3 hours. After this time, 10 mL of water was added to the flask, the mixture was transferred to a separatory funnel. and extracted three times with ethyl acetate (10 ml each) The organic phases were combined, dried over anhydrous MgSO4 and purified by flash chromatography (S1O2, hexane / ethyl acetate, 4/1) to give the product 4.2 in 97% yield (0.57) g).

¹ H NMR (600 MHz, CDCl 3) δ [ppm]: 7.56 (d, J = 7.3 Hz, 1H), 7.43 (d, J = 7.3 Hz, 1H), 7.31 ( s, 1H), 4.88 (s, 2H), 4.75 (s, 2H), 1.35 (s, 12H) ¹³ C NMR (151 MHz, CDCl 3) δ [ppm]: 153.8; 133.4; 130.1; 128.7; 117.0; 115.2; 84.3; 61.0; 53.9; 25.0

End product 4. 2 - ((3 '- (benzo-1,4-dioxan-6-yl) -2 ^, -chloro -) - 4- (hydroxymethyl) - [1, rbiphenyl] -3-yl) oxy) acetonitrile

(2.2)

dioxane, water

Pd (iippf) CL ₂ * DCM K ₂ CO ₅

(4)

6- (3-Bromo-2-chlorophenyl) benzo-1,4-dioxane (0.40 g, 1.24 mmol, 1.0 eq.), (2- (2-hydroxymethyl) - 5- (4,4,5,5-tetramethyl-1,3,2-dioxyborolan-2-yl) phenoxy) acetonitrile (0.50 g, 1.72 mmol, 1.4 eq.), K2CO3 (0, 51 g, 3.72 mmol, 3.0 eq.), And a dioxane / water mixture (15 ml, 2/1, v, v). The solution thus obtained was deoxygenated for 30 minutes via

PL241 191 Β1 argon wash followed by the addition of Pd (dppf) ChDCM (0.10 g, 0.12 mmol, 0.1 eq.). The contents of the flask were heated at 80 ° C for 5 hours under an inert gas atmosphere. At the end of this time, the contents of the flask were transferred to a separating funnel and extracted three times with ethyl acetate (20 ml each). The organic phases were combined, dried over anhydrous MgSO4 and purified by flash chromatography (S1O2, hexane / ethyl acetate, 1/1). The product 3 was obtained in a yield of 56% (0.28 g).

¹ H NMR (600 MHz, CDCl 3) δ [ppm]: 7.49 (d, J = 7.7 Hz, 1H), 7.36-7.32 (m, 2H), 7.29 (dd, J = 6.5, 2.8 Hz, 1H), 7.19 (dd, J = 7.7, 1.4 Hz, 1H), 7.06 (d, J = 1.3 Hz, 1H), 7.00 -6.99 (m, 1H), 6.96-6.91 (m, 2H), 4.86 (s, 2H), 4.78 (s, 2H), 4.31 (s, 4H), 1.90 (s, 1H) ¹³ C NMR (151 MHz, CDCl 3) δ [ppm]: 153.8; 143.4; 143.2; 141.4; 141.3; 133.2; 131.0; 130.2

Example 5. (3 ^, - (benzo-1,4-dioxan-6-yl) -2'-chloro-3- (cyclobutyl) methoxy- [1,1-biphenyl] -4-yl) methanol.

Intermediate product 5.1. 4-bromo-2- (cyclobutylmethyloxy) benzyl alcohol

(3-1)

CsjCOj

DMF anhydrous

(5-1)

4-bromo-2-hydroxybenzyl alcohol (1.00 g, 4.90 mmol, 1.0 eq.), (Bromomethyl) cyclobutane (1.10 ml, 9.80 mmol, 2.0 eq.) Was placed in a round bottom flask. ), K2CO3 (1.35 g, 9.80 mmol, 2.0 eq.) And anhydrous DMF (20 ml). The resulting mixture was left on a magnetic stirrer overnight and then the reaction progress was checked by TLC (S1O2, hexane / ethyl acetate, 4/1, v / v). After complete conversion of the alcohol, the mixture was transferred to a separating funnel, added with 10 ml of water and extracted three times with ethyl acetate (15 ml each). The organic layers were combined, dried over anhydrous MgSO4, and the solvent was evaporated. The product was purified by flash column chromatography (hexane / ethyl acetate, 4/1, v / v) to give compound 5.1 in 59% yield (0.79 g).

¹ H NMR (600 MHz, CDCl 3) δ [ppm]: 7J4 (d, 'J = 8.0 Hz, 1H), 7.07' (dd, J = 7.9; 1.8 Hz, 1H), 7.00 (d, J = 1.8 Hz, 1H), 4.63 (s, 2H), 3.96 (d, J = 6.6 Hz, 2H), 2.83-2.75 (m, 1H ), 2.18-2.12 (m, 2H), 2.041.92 (m, 2H), 1.90-1.83 (m, 2H) ^13C NMR (151 MHz, CDCl3) δ [ppm]: 157.7; 129.8; 128.5; 123.6; 122.1; 114.9; 72.4; 61.8; 34.5; 24.9; 18.7

Intermediate product 5.2. (2-ethoxy-4- (4,4,5,5-tetramethyl-1,3,2-dioxyborolan-2-yl) phenyl) methanol

(5-1)

WfpOCI / dcm AcOK anhydrous dioxane

(5.2)

In a two-necked round bottom flask was placed bis (pinacolato) diborane (0.81 g, 3.19 mmol, 1.1 eq), anhydrous potassium acetate (0.85 g, 8.70 mmol, 3.0 eq), 4-bromo alcohol -2- (cyclobutylmethoxy) benzyl (0.78 g, 2.90 mmol, 1.0 eq,) and anhydrous dioxane (20 mL). The solution thus obtained was deoxygenated for 30 minutes by purging with argon, then Pd (dppf) ChDCM (0.07 g, 0.09 mmol, 0.03 eq,) was added. The contents of the flask were heated at 80 ° C for 3 hours. After this time, 10 ml of water was added to the flask, the mixture was transferred to a separating funnel and extracted three times with ethyl acetate (15 ml each). The organic phases were combined, dried over anhydrous MgSO4 and purified by flash chromatography (S1O2, hexane / ethyl acetate, 4/1). The product 5.2 was obtained in a yield of 72% (0.66 g).

¹ H NMR (600 MHz, CDCl 3) δ [ppm]: 7.40 (d, J = 7.4 Hz, 1H), 7.28-7.26 (m, 2H), 4.70 (s, 2H ), 4.06 (d, J = 6.6Hz, 2H), 2.84-2.76 (m, 1H), 2.19-2.11 (m, 2H), 2.03-1.84 (m, 4H). 35 (s, 12H) ¹³ C NMR (151 MHz, CDCl 3) δ [ppm]: 156.8; 132.5; 128.1; 127.6; 116.7; 84.0; 72.2; 62.8; 34.8; 29.8; 25.0; 25.0; 18.8.

End product 5. (3 ^, - (benzo-1,4-dioxan-6-yl) -2'-chloro-3- (cyclobutyl) methoxy- [1,1-biphenyl] -4-yl) methanol

PL241 191 Β1

P ^ dppęCL / DCM KjCOj dioxane in water

6- (3-Bromo-2-chlorophenyl) benzo-1,4-dioxane (0.49 g, 1.50 mmol, 1.0 eq.), (2- (cyclobutylmethoxy) -4- (4,4,5,5-tetramethyl-1,3,2-dioxyborolan-2-yl) phenyl) methanol (0.67 g, 2.10 mmol, 1.4 eq.), K2CO3 (0.62 g , 4.48 mmol, 3.0 eq.), And a dioxane / water mixture (20 mL, 2/1, v, v). The solution thus obtained was deoxygenated for 30 minutes by purging with argon, then Pd (dppf) Cl2-DCM (0.12 g, 0.15 mmol, 0.1 eq.) Was added. The contents of the flask were heated at 80 ° C for 5 hours under an inert gas atmosphere. At the end of this time, the contents of the flask were transferred to a separating funnel and extracted three times with ethyl acetate (20 ml each). The organic phases were combined, dried over anhydrous MgSO4 and purified by flash chromatography (S1O2, hexane / ethyl acetate, 1/1). The product 5 was obtained in a yield of 65% (0.42 g).

¹ H NMR (600 MHz, CDCl 3) δ [ppm]: 7.34-7.27 (m, 4H), 7.02-698 (m, 3H), 6.96-6.92 (m, 2H) , 4.75 (s, 2H), 4.31 (s, 4H), 4.02 (d, J = 6.6Hz), 2H), 2.87-2.78 (m, 1H), 2 , 19-2.12 (m, 2H), 2.02-1.85 (m, 4H) ¹³ C NMR (151 MHz, CDCl 3) δ [ppm]: 156.7; 143.3; 143.1; 141.5; 141.2; 141.1; 133.5; 131.1; 130.7; 130.2; 128.6; 128.4; 126.5; 122.9; 121.8; 118.7; 117.0; 113.0; 72.2; 64.6; 64.6; 62.5; 34.7; 25.0; 18.8

Example 6, 3 ^, - (Benzo-1,4-dioxan-6-yl) -2'-chloro-4- (hydroxymethyl) - [1,1-biphenyl] -3-yl-dimethylcarbamate

Intermediate 6.1.5-Bromo-2- (hydroxymethyl) phenyl dimethyl carbamate (3.1)

ACN anhydrous (3.1)

KfCOg pyridine

(6.1)

4-bromo-2-hydroxybenzyl alcohol (0.50 g, 2.46 mmol, 1.0 eq.), Pyridine (2.50 ml) and anhydrous ACN (8 ml) were placed in a round bottom flask under an inert gas atmosphere (Ar). ). The contents of the flask were stirred until the alcohol dissolved, then K2CO3 (0.68 g, 4.93 mmol, 2.0 eq.) Was added and stirred for 20 minutes. After this time, a solution of N, N-dimethylcarbamoyl chloride (0.46 mL, 4.92 mmol, 2 eq.) In 5 mL anhydrous ACN was added dropwise to the flask. The resulting solution was left on a magnetic stirrer for 2 hours and then the progress of the reaction was checked by TLC (S1O2, hexane / ethyl acetate, 1/1, v / v). After complete conversion of the alcohol, the mixture was transferred to a separating funnel, added with 10 ml of water and extracted three times with ethyl acetate (15 ml each). The organic layers were combined, dried over anhydrous MgSO 4, and the solvent was evaporated. The product was purified by flash column chromatography (hexane / ethyl acetate, 1/1, v / v) to give compound 6.1 in 97% yield (0.65 g).

¹ H NMR (600 MHz, CDCl 3) δ [ppm]: 7.03 (m, 1H), 6.91 (m, 2H), 3.52 (s, 2H), 2.21 (s, 6H) ¹³ CNMR (151 MHz, CDCl3) δ [ppm]: 158.4; 130.7; 122.5; 121.3; 120.4; 117.9; 59.5; 44.2 Intermediate product 6.2. 2- (hydroxymethyl) -5- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) phenyl dimethyl carbamate.

Pd (dppf) Cl ₃ * DCM AcOK dioxane anhydrous (6.1)

In a two-necked round bottom flask was placed bis (pinacolato) diborane (0.57 g, 2.26 mmol, 1.2 eq), anhydrous potassium acetate (0.55 g, 5.65 mmol, 3.0 eq), 5- bromo-2- (hydroxymethyl) phenyl dimethyl carbamate (0.52 g, 1.88 mmol, 1.0 eq,) and anhydrous dioxane (15 ml). The solution thus obtained was deoxygenated for 30 minutes by purging with argon, then Pd (dppf) Cl2-DCM (0.08 g, 0.09 mmol, 0.05 eq,) was added. The contents of the flask were heated at 80 ° C for 3 hours. After this time, 10 ml of water was added to the flask, the mixture was transferred to a separating funnel and extracted three times with ethyl acetate (10 ml each). The organic phases were combined, dried over anhydrous MgSO4 and purified

PL241 191 Β1 using flash chromatography (S1O2, hexane / ethyl acetate, 1/2). The product 6.2 was obtained in a yield of 77% (0.46 g).

¹ H NMR (600 MHz, CDCl 3) δ [ppm]: 7.67 (dd, J = 7.5; 1.0 Hz, 1H), 7.47 (m, 2H), 4.56 (s, 2H ), 3.12 (s, 3H), 3.01 (s, 3H), 1.33 (s, 12H), 1.22 (s, 1H) ^13C NMR (151 MHz, CDCl3) δ [ppm] : 155.9; 149.1; 136.6; 132.8; 129.9; 128.5; 84.1; 60.6; 37.0; 36.7; 25.0

End product 6. 3 ^, - (Benzo-1,4-dioxan-6-yl) -2 ^, -chloro-4- (hydroxymethyl) - [1,1-biphenyl] -3-yl dimethyl carbamate.

(2-2)

diocian / water

Wppf) Ci ₂ * r> CM KiCOj

(6)

6- (3-Bromo-2-chlorophenyl) benzo-1,4-dioxane (0.57 g, 1.74 mmol, 1.2 eq.), 2- (hydroxymethyl) -5- ( 4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) phenyl dimethyl carbamate (0.46 g, 1.45 mmol, 1.0 eq.), K2CO3 (0.80 g, 5 , 78 mmol, 4.0 eq.), And a dioxane / water mixture (15 mL, 2/1, v, v). The solution thus obtained was deoxygenated for 30 minutes by purging with argon, then Pd (dppf) Cl2-DCM (0.06 g, 0.07 mmol, 0.05 eq.) Was added. The contents of the flask were heated at 80 ° C for 5 hours under an inert gas atmosphere. At the end of this time, the contents of the flask were transferred to a separating funnel and extracted three times with ethyl acetate (20 ml each). The organic phases were combined, dried over anhydrous MgSO4 and purified by flash chromatography (SiO2, hexane / ethyl acetate, 1/1). The product 6 was obtained in a yield of 30% (0.19 g).

¹ H NMR (600 MHz, CDCl 3) δ [ppm]: 7.30-7.27 (m, 3H), 7.01-6.99 (m, 2H), 6.95-6.91 (m, 3H), 6.87 (dd, J = 7.6; 1.6 Hz, 1H), 4.30 (s, 4H), 3.70 (s, 2H), 2.37 (s, 6H).

¹³ C NMR (151 MHz, CDCl 3) δ [ppm]: 157.8; 143.2; 143.1; 141.6; 141.1; 140.9; 133.7; 131.1; 130.4; 130.3; 127.9; 126.4; 123.0; 121.2; 120.4; 118.7; 117.4; 116.9; 64.6; 64.5; 62.9; 44.8

Example 7. 1 - ((3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1,1-biphenyl] -4-yl) methyl) piperazine

In a round bottom flask was placed (3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1,1'-biphenyl] -4-yl) methanol (0.15 g , 0.39 mmol, 1.0 eq.) Under an inert gas atmosphere (Ar) and dissolved in 5 ml of anhydrous dichloromethane. The resulting mixture was cooled in an ice bath before SOCl2 (0.14 mL, 1.95 mmol, 5.0 eq.) Was added dropwise without bringing the solvent to boiling, then anhydrous DMF (0.04 mL) was added in one portion. The flask contents were stirred for 2 hours and the reaction progress was checked by TLC (S1O2, hexane / ethyl acetate, 4/1). After the starting material had completely disappeared, saturated NaHCO 3 solution was added dropwise to the flask until gas evolution ceased. The mixture was transferred to a separatory funnel, 5 ml of saturated NaHCO 3 was added and the aqueous layer was extracted three times with DCM (10 ml each). The organic layers were combined, dried over anhydrous MgSO4, and the solvent was evaporated.

The oil obtained in the previous step was placed in a round bottom flask under an inert gas (Ar) atmosphere, 1-Boc-piperazine (0.36 g, 1.95 mmol, 5.0 eq.) And anhydrous THF (5 ml) were added and left to stand in the atmosphere. stirrer for 48 h. After this time, the contents of the flask were transferred to a separatory funnel with saturated Na2CO3 solution (10 mL) and the aqueous layer was washed three times with ethyl acetate (10 mL each). The organic phases were combined, dried over anhydrous MgSO4 and the solvent was evaporated. The product was purified using flash column chromatography (S1O2, hexane / ethyl acetate, 2/1). The purified product was deprotected with the amino group. For this, it was dissolved in anhydrous dioxane, then a 4M solution of HCl in dioxane (0.98 ml, 3.90 mmol, 10.0 eq.) Was added and left on a magnetic stirrer overnight. The next day, the contents of the flask were transferred to a separatory funnel with saturated Na2CO3 solution (7 mL). The aqueous layer was extracted three times with ethyl acetate (10 ml each), the organic phases were combined, dried over anhydrous MgSO4 and evaporated

PL241 191 Β1 solvent, the product with the deprotected amino group was purified by column chromatography (methanol / 7M NH3 in methanol, 20/1, v / v) to give compound 7 as a colorless solid in a yield of 33% (0.06 g).

¹ H NMR (600 MHz, DMSO-d6) δ [ppm]: 7.43 (m, 1H), 7.38 (m, 2H), 7.35 (dd, J = 7.5; 1.5 Hz , 1H), 7.02 (s, 1H), 6.99 (d, J = 7.6 Hz, 1H), 6.95-6.93 (m, 2H), 6.90 (dd, J = 8.3; 1.8 Hz, 1H), 4.29 (s, 4H), 3.80 (s, 3H), 3.46 (s, 2H), 2.70 (t, J = 54.43 Hz, 1H), 2.35 (m, 5H).

¹³ C NMR (151 MHz, DMSO-d6) δ [ppm]: 157.3; 143.6; 143.3; 141.5; 140.9; 139.6; 133.0; 130.9;

130.8; 130.4; 129.8; 127.4; 125.9; 122.9; 121.6; 118.5; 117.2; 112.5; 64.6; 64.6; 56.5; 55.9; 54.9; 46.2

Example 8. 1-carboxamide 4 - ((3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1,1-biphenyl] -4-yl) methyl) piperazines

DCM anhydrous

In a round bottom flask was placed (3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1,1'-biphenyl] -4-yl) methanol (0.15 g , 0.39 mmol, 1.0 eq.) Under an inert gas atmosphere (Ar) and dissolved in 5 ml of anhydrous dichloromethane. The resulting mixture was cooled in an ice bath before SOCl2 (0.14 mL, 1.95 mmol, 5.0 eq.) Was added dropwise without bringing the solvent to boiling, then anhydrous DMF (0.04 mL) was added in one portion. The flask contents were stirred for 2 hours and the reaction progress was checked by TLC (S1O2, hexane / ethyl acetate, 4/1). After the starting material had completely disappeared, saturated NaHCO 3 solution was added dropwise to the flask until gas evolution ceased. The mixture was transferred to a separatory funnel, 5 ml of saturated NaHCO 3 was added and the aqueous layer was extracted three times with DCM (10 ml each). The organic layers were combined, dried over anhydrous MgSO4, and the solvent was evaporated.

The oil obtained in the previous step was placed in a round bottom flask under an inert gas (Ar) atmosphere, 1-carboxamidopiperazine hydrochloride (0.13 g, 0.78 mmol, 2.00 eq.) And anhydrous THF (5 ml) were added. To the mixture thus prepared was added EtsN (0.11.0.78 mmol, 2.00 eq.) And heated at 50 ° C overnight. At the end of this time, the contents of the flask were transferred to a separatory funnel with saturated Na2CO3 solution (5 mL) and the aqueous layer was washed three times with ethyl acetate (10 mL each). The organic phases were combined, dried over anhydrous MgSO4 and the solvent was evaporated. The product was purified using flash column chromatography (SiO2, hexane / ethyl acetate, 1/1) to provide compound 6 as a colorless solid in a yield of 36% (0.07 g).

¹ H NMR (600 MHz, CDCl 3) δ [ppm]: 7.31 (d, J = 7.7 Hz, 1H), 7.24-7.17 (m, 3H), 6.943 (dd, J = 7 , 7, 1.5 Hz, 1H), 6.91 (d, J = 1.9 Hz, 1H), 6.89 (d, J = 1.5 Hz, 1H), 6.86-76.83 (m, 2H), 4.71 (s, 2H), 4.20 (s, 4H), 3.74 (s, 3H), 3.54 (s, 2H), 3.34 (t, J = 5.0 Hz, 4H), 2.45 (t, J = 5.1 Hz, 4H).

¹³ C NMR (151 MHz, CDCl 3) δ [ppm]: 158.3; 157.2; 143.2; 143.0; 141.5; 141.1; 140.2; 133.4; 130.9; 130.5; 130.2; 130.0; 126.4; 125.0; 122.8; 121.6; 118.6; 116.9; 112.1; 64.5; 64.4; 56.0; 55.6; 52.8; 44.1

Example 9. 1- (4 - ((3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1,1-biphenyl] -4-yl) methyl) piperazin-1-yl) ethane-1-one

DMF anhydrous

In a round bottom flask was placed (3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1,1'-biphenyl] -4-yl) methanol (0.15 g , 0.39 mmol, 1.0 eq.) Under an inert gas atmosphere (Ar) and dissolved in 5 ml of anhydrous dichloromethane. The resulting mixture was cooled in an ice bath before SOCl2 (0.14 mL, 1.95 mmol, 5.0 eq.) Was added dropwise without bringing the solvent to boiling, then anhydrous DMF (0.04 mL) was added in one portion. The flask contents were stirred for 2 hours and the reaction progress was checked by TLC (S1O2, hexane / ethyl acetate, 4/1). After the starting material had completely disappeared, saturated NaHCO 3 solution was added dropwise to the flask until gas evolution ceased. The mixture was transferred to a separatory funnel, 5 ml of saturated NaHCO 3 was added and the aqueous layer was extracted three times with DCM (10 ml each). The organic layers were combined, dried over anhydrous MgSO4, and the solvent was evaporated.

PL241 191 Β1

The oil obtained in the previous step was placed in a round bottom flask under an inert gas (Ar) atmosphere, 1-acetylpiperazine (0.10 g, 0.78 mmol, 2.00 eq.) And anhydrous DMF (5 ml) were added. To the mixture thus prepared was added DIPEA (0.14, 0.78 mmol, 2.00 eq.) And heated at 80 ° C overnight. At the end of this time, the contents of the flask were transferred to a separatory funnel with saturated Na2CO3 solution (5 mL), and the aqueous layer was washed three times with ethyl acetate (10 mL each). The organic phases were combined, dried over anhydrous MgSO4 and the solvent was evaporated. The product was purified using flash column chromatography (S1O2, hexane / ethyl acetate, 1/1) to provide compound 7 as a colorless solid in a yield of 76% (0.15 g).

¹ H NMR (600 MHz, CDCl 3) δ [ppm]: 7.30 '(d, J = 7.7 Hz, 1H), 7.23-7.17 (m, 3H), 6.92 (dd, J = 7.6; 1.5 Hz, 1H), 6.91 (d, J = 1.9 Hz, 1H), 6.88 (d, J = 1.5 Hz, 1H), 6.85- 6.82 (m, 2H), 4.18 (s, 4H), 3.74 (s, 3H), 3.56 (t, J = 4.9 Hz, 2H), 3.54 (s, 2H ), 3.39 (t, J = 4.8 Hz, 2H), 2.44 (m, 4H), 1.99 (s, 3H) ^13C NMR (151 MHz, CDCl3) δ [ppm]: 169 0; 157.3; 143.2; 143.0; 141.5; 141.1; 140.3; 133.3; 130.9; 130.5; 130.2; 130.1; 126.4; 124.7; 122.8; 121.6; 118.6; 116.9; 112.1; 64.4; 64.4; 55.8; 55.5; 53.1; 52.7; 46.4; 41.5; 21.4

Example 10. 2-Amino-1- (4 - ((3 '- (benzo-1,4-dioxan-6-yl) -2 ^, -chloro-3-methoxy- [1,1-biphenyl] -4-) yl) methyl) piperazin-1-yl) ethan-1-one

Intermediate product 10.1. Tert-Butyl (2-oxo-2- (piperazin-1-yl) ethyl) carbamate

N-Boc-glycine (1.00 g, 5.71 mmol, 1.0 eq.), HATU (2.17 g, 5.71 mmol, 1.0 eq.) And anhydrous DMF (20 mL) were placed in a round bottom flask. . The contents of the flask were stirred for 20 minutes on a magnetic stirrer, then piperazine (1.48 g, 17.13 mmol, 3 eq.) Was added and left on the stirrer for 2 days. After this time, the solvent was azeotroped (toluene). The product was purified using column chromatography (SiO2, methanol) to afford intermediate 10.1 in 55% yield (0.76 g).

¹ HNMR (600 MHz, CDCl 3) δ [ppm]: 5.61 (s, 1H), 3.96 (s, 2H), 3.64 (s, 2H), 3.42 (s, 2H), 2 , 91 (s, 4H), 1.44 (s, 9H) 1 ¹³ C NMR (151 MHz, CDCl 3) δ [ppm]: 167.0; 155.9; 79.6; 45.6; 45.3; 45.0; 42.4; 42.2; 28.4

End product 10. 2-amino-1- (4 - ((3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy [1,1'-biphenyl] -4) -yl) methyl) piperazin-1-yl) ethan-1-one

(1)

DCM anhydrous

(10.1)

1. DMFHzwOne

2. Dioxati anhydrous

1. DIPEA

2. 4M EC1 in dioxane

In a round bottom flask was placed (3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1,1'-biphenyl] -4-yl) methanol (0.15 g , 0.39 mmol, 1.0 eq.) Under an inert gas atmosphere (Ar) and dissolved in 5 ml of anhydrous dichloromethane. The resulting mixture was cooled in an ice bath before SOCl2 (0.14 mL, 1.95 mmol, 5.0 eq.) Was added dropwise without bringing the solvent to boiling, then anhydrous DMF (0.04 mL) was added in one portion. The flask contents were stirred for 2 hours and the reaction progress was checked by TLC (S1O2, hexane / ethyl acetate, 4/1). After the starting material had completely disappeared, saturated NaHCO 3 solution was added dropwise to the flask until gas evolution ceased. The mixture was transferred to a separatory funnel, 5 ml of saturated NaHCO 3 was added and the aqueous layer was extracted three times with DCM (10 ml each). The organic layers were combined, dried over anhydrous MgSO4, and the solvent was evaporated.

The oil obtained in the previous step was placed in a round bottom flask under an inert gas (Ar) atmosphere, tert-butyl (2-oxo-2- (piperazin-1-yl) ethyl) carbamate (0.19 g, 0.78 mmol, 2-oxo-2- (piperazin-1-yl) ethyl) was added. 0 eq.) And anhydrous DMF (5 ml). To the mixture so prepared was added DIPEA (0.14 mL, 0.78 mmol, 2.0 eq.) And heated at 80 ° C overnight. At the end of this time, the contents of the flask were transferred to a separatory funnel with saturated Na2CO3 solution (5 mL) and the aqueous layer was washed three times with ethyl acetate (10 mL each). The organic phases were combined, dried over anhydrous MgSO4 and the solvent was evaporated. The obtained paste was dissolved in anhydrous dioxane (5 mL), 4M HCl was added

PL241 191 Β1 in dioxane (1.00 mL, 4.00 mmol, 10.0 eq.) And left on the stirrer overnight. The next day, the contents of the flask were transferred to a separatory funnel with saturated Na2CO3 solution (5 mL). The aqueous layer was extracted three times with ethyl acetate (10 ml each), the organic phases were combined, dried over anhydrous MgSO4 and the solvent was evaporated. The amino deprotected product was purified by column chromatography (S1O2, methanol / 7M NH3 in methanol, 20/1, v / v) to give compound 10 as a colorless solid in 20% yield (0.04 g).

¹ HNMR (600 MHz, CDCl3) δ [ppm]: 7.30 (d, 'J = 7.7 Hz, 1H), 7.25-7.19 (m, 3H), 6.94-6.92 (m, 2H), 6.89 (m, 1H), 6.87-6.83 (m, 2H), 4.22 (s, 4H), 3.75 (s, 3H), 3.59 ( m, 2H), 3.54 (s, 2H), 3.48 (wide, 2H), 3.34 (m, 2H), 3.05 (wide, 2H), 2.45 (m, 4H) ¹³ C NMR (151 MHz, CDCl 3) δ [ppm]: 170.2; 157.2; 143.2; 143.0; 141.5; 141.1; 140.3; 133.3; 130.9; 130.5; 130.2; 130.0; 126.3; 124.8; 122.8; 121.6; 118.6; 116.9; 112.1; 64.5; 64.4; 55.9; 55.6; 52.9; 52.8; 44.3; 42.7; 42.2

Example 11. 1 - ((3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1,1'-biphenyl I] -4-yl) methyl ) -4- (methylsulfonyl) piperazine

The procedure described in Example 9 was used to prepare compound 11.

(2)

(U) (3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1, T-biphenyl] -4-yl) methanol (0.15 g, 0 , 39 mmol, 1.0 eq.), SOCI2 (0.14 mL, 1.95 mmol, 5.0 eq.), Anhydrous DMF (0.04 mL),

1- (methylsulfonyl) piperazine (0.13 g, 0.78 mmol, 2.0 eq.), DIPEA (0.14 ml, 0.78 mmol, 2.0 eq.) Product 11 was obtained in a yield of 89% (0, 18 g).

¹ H NMR (600 MHz, CDCl 3) δ [ppm]: 7.39 (d, J = 7.7 Hz, 1H), 7.36-7.31 (m, 3H), 7.03 (dd, J = 7.6; 1.5 Hz, 1H), 7.02 (d, J = 1.8 Hz, 1H), 7.00 (d, J = 1.4 Hz, 1H), 6.97-6 , 94 (m, 2H), 4.32 (s, 4H), 3.86 (s, 3H), 3.68 (s, 2H), 3.30 (m, 4H), 2.80 (s, 3H), 2.69 (t, J = 4.2 Hz, 4H) ¹³ C NMR (151 MHz, CDCl 3) δ [ppm]: 157.3; 143.2; 143.0; 141.4; 141.1; 140.4; 133.3; 130.9; 130.5; 130.1; 130.0; 126.4; 124.7; 122.8; 121.6; 118.6; 116.9; 112.2; 64.5; 64.4; 55.7; 55.6; 52.3; 46.0; 34.1

Example 12. / V- (2 - (((3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1, r-biphenyl] -4-yl) methyl) amino) ethyl) acetamide

The procedure described in Example 9 was followed to prepare compound 12.

(3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1, T-biphenyl] -4-yl) methanol (0.15 g, 0.39 mmol , 1.0 eq.), SOCI2 (0.14 mL, 1.95 mmol, 5.0 eq.), Anhydrous DMF (0.04 mL), N - (2-aminoethyl) acetamide (0.19 mL , 1.95 mmol, 5.0 eq.), DIPEA (0.14 mL, 0.78 mmol, 2.0 eq.). The product 12 was obtained in a yield of 28% (0.05 g). Column chromatography: S1O2, chloroform / 7M NH3 in methanol, 20/1, v / v.

¹ HNMR (600 MHz, DMSO-d6) δ [ppm]: 7.33 (m, 4H), 7.00-6.98 (m, 3H), 6.95-6.91 (m, 2H), 6.33 (s, 1H), 4.29 (s, 4H), 3.86 (s, 3H), 3.81 (s, 2H), 3.36 (m, 2H), 2.78 (t , J = 5.8 Hz, 2H), 2.00 (Wide, 1H), 1.98 (s, 3H).

¹³ C NMR (151 MHz, DMSO-d6) δ [ppm]: 170.3; 157.1; 143.2; 143.0; 141.4; 141.1; 140.5; 133.3; 130.9; 130.5; 130.1; 129.5; 127.2; 126.4; 122.8; 121.6; 118.6; 116.9; 112.0; 64.5; 64.4; 55.4; 48.7; 47.9; 39.1; 23.3

Example 13. 2-Amino- / V- (2 - ((3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1,1-biphenyl] -4) -yl) methyl) amino) ethyl) acetamide

Intermediate product 13.1. Tert-butyl (2 - ((2-aminoethyl) amino) -2-oxoethyl) carbamate

ii ₂ n nh ₂

McOH / CHCl ₃ 10/1

(13.1)

PL241 191 Β1

In a two-neck round bottom flask under an inert gas atmosphere (Ar) was charged tert-butyl 2 - ((2-aminoethyl) amino) -2-oxoethyl) carbamate (1.00 g, 5.28 mmol, 1 eq.) And dissolved in methanol / water (33 ml, 10/1, v / v). The contents of the flask were cooled in an ice bath, then a methanolic solution of ethylenediamine (1.77 mL, 26.43 mmol, 5.0 eq. In 15 mL MeOH) was added dropwise. The contents of the flask were left on the stirrer for 3 days, then the solvent was evaporated under reduced pressure. The product was purified by column chromatography (S1O2, methanol / 7M NH3 in methanol, 20/1) to give the product 13.1 in 48% yield (0.55 g).

¹ HNMR (600 MHz, DMSO-ds) δ [ppm]: 6.79 (Wide, 1H), 5.42 (Wide, 1H), 3.78 (d, J = 5.6 Hz, 2H), 3 , 77 (m, 2H), 2.82 (t, J = 5.7 Hz, 2H), 1.44 (s, 9H) ¹³ C NMR (151 MHz, DMSO-d ₆ ) δ [ppm]: 167 0; 156.2; 80.3; 44.4; 41.8; 41.2; 28.3

End product 13. 2-amino-N- (2 - ((3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1,1 ^, -biphenyl] - 4-yl) methyl) amino) ethyl) acetamide

The procedure described in Example 10 was used to prepare compound 13.

□ CM anhydrous

l.DCPEA

1.4MMC1 in the diakuttle

1. DMF anhydrous and dsokun anhydrous

(13) (3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1,1'-biphenyl] -4-yl) methanol (0.15g, 0 , 39 mmol, 1.0 eq.), SOCI2 (0.14 mL, 1.95 mmol, 5.0 eq.), Anhydrous DMF (0.04 mL), (2 - ((2-aminoethyl) amino) -2 tert-butyl oxoethyl) carbamate (0.17 g, 0.78 mmol, 2.0 eq.), DIPEA (0.14 mL, 0.78 mmol, 2.0 eq.), 4M HCl in dioxane (1 0.00 mL, 4.0 mmol, 10.0 eq.). Compound 13 was obtained in a yield of 43.3% (0.08 g).

¹ H NMR (600 MHz, CDCl 3) δ [ppm]: 7.47 (s, 1H), 7.22 (m, 4H), 6.91-6.85 (m, 5H), 4.22 (s , 4H), 3.77 (s, 5H), 3.38-3.38 (m, 4H), 2.74 (s, 2H) ¹³ C NMR (151 MHz, CDCl 3) δ [ppm]: 172. 8; 157.1; 143.2; 143.0; 141.4; 141.1; 140.4; 133.3; 130.9; 130.5; 130.1; 129.5; 126.4; 122.8; 121.6; 118.6; 116.8; 112.0; 64.5; 64.4; 55.4; 48.5; 48.0: 44.9: 38.6

Example 14. W- (2 - (((3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1,1-biphenyl] -4-yl) methyl ) amino) ethyl) methanesulfonamide

The procedure described in Example 9 was followed to prepare compound 14.

(3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1,1'-biphenyl] -4-yl) methanol (0.15 g, 0.39 mmol, 1.0 eq.), SOCI2 (0.14 ml, 1.95 mmol, 5.0 eq.), anhydrous DMF (0.04 ml), N - (2-aminoethyl) methanesulfamide (0.27 mL, 1.95 mmol, 5.0 eq.), DIPEA (0.14 mL, 0.78 mmol, 2.0 eq.). The product 14 was obtained in 89% yield (0.18 g). Column chromatography: SiO2, chloroform / 7M NH3 in methanol, 20/1, v / v.

¹ H NMR (600 MHz, CDCl 3) δ [ppm]: 7.26-7.21 (m, 3H), 7.19 (dd, J = 6.6; 2.3 Hz, 1H), 6.94 -6.91 (m, 3H), 6.87-6.84 (m, 2H), 4.85 (Wide, 2H), 4.22 (s, 4H), 3.83 (s, 2H), 3.81 (s, 3H), 3.22 (t, J = 5.7 Hz, 2H), 2.87 (s, 3H), 2.84 (t, J = 5.6 Hz, 2H) ¹³ C NMR (151 MHz, CDCl 3) δ [ppm]: 157.2; 143.2; 143.0; 141.4; 141.1; 141.1; 133.2; 130.9; 130.7; 130.1; 126.4; 124.5; 122.8; 121.8; 118.6; 116.9; 112.2; 64.5; 64.4; 55.6; 48.1; 47.6; 41.7; 40.0

Example 15. 1- (2 - (((3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1,1-biphenyl] -4-yl) methyl) ) amino) ethyl) urea

The procedure described in Example 8 was followed to prepare compound 15.

(1S>

(3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1,1'-biphenyl] -4-yl) methanol (0.15 g, 0.39 mmol, 1.0 eq.), SOCI2 (0.14 ml, 1.95 mmol, 5.0 eq.), anhydrous DMF (0.04 ml), 1- (2-aminoethyl) urea hydrochloride (0.11 ml, 1.95 mmol, 5.0 eq.), EtsN (0.11 ml, 0.78 mmol, 2.0 eq.). The product 15 was obtained in a yield of 55% (0.10 g). Column chromatography: SiO2, chloroform / 7M NH3 in methanol, 20/1, v / v.

PL241 191 Β1 ¹ NMR (600 MHz, CDCl ₃ ) δ [ppm]: 7.33-7.27 (m, 3Η), 7.24 (d, J = 7.5 Hz, 1H), 6.99 -6.96 (m, 3H), 6.94-6.91 (m, 2H), 5.70 (Wide, 1H), 4.93 (Wide, 2H), 4.29 (s, 4H), 3.84 (s, 3H), 3.80 (s, 2H), 3.27 (m, 2H), 2.77 (t, J = 5.5 Hz, 2H), 2.19 (Wide, 1H ) ¹³ C NMR (151 MHz, CDCl ₃ ) δ [ppm]: 159.7; 157.1; 143.2; 143.0; 141.4; 141.1; 140.6; 133.3; 130.9; 130.5; 130.1; 129.5; 126.9; 126.4; 122.8; 121.6; 118.6; 116.9; 112.0; 64.5; 64.4; 55.4; 48.9; 48.8; 40.3

Example 16 2 - (((3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [11,1'-biphenyl] -4-yl) methyl) amino ) propane-1,3-diol

The procedure described in Example 9 was followed to prepare compound 16.

(3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1,1'-biphenyl] -4-yl) methanol (0.15g, 0.39mmol , 1.0 eq.), SOCI2 (0.14 mL, 1.95 mmol, 5.0 eq.), Anhydrous DMF (0.04 mL), serinol (0.07 mL, 1.95 mmol, 5, 0 eq.), DIPEA (0.09 mL, 0.78 mmol, 2.0 eq.). The product 16 was obtained in a yield of 68% (0.12 g). Column chromatography: S1O2, chloroform / 7M NH3 in methanol, 20/1, v / v.

¹ H NMR (600 MHz, CDCl 3) δ [ppm]: 7.26 (d, J = 7.6 Hz, 1H), 7.23-7.21 (m, 2H), 7.18 (m, 1H ), 6.93-6.90 (m, 3H), 6.86-6.83 (m, 2H), 4.84 (Wide, 2H), 4.22 (s, 4H), 3.98 ( s, 2H), 3.80 (s, 3H), 3.72 (m, 2H), 3.61 (m, 2H), 2.86 (m, 1H), 1.84 (s, 1H) ¹³ C NMR (151 MHz, CDCl 3) δ [ppm]: 157.2; 143.2; 143.0; 141.8; 141.1; 141.1; 133.2; 130.9; 130.7; 130.3; 130.1; 126.4; 123.3; 122.8; 122.0; 118.6; 116.9; 112.2; 77.1; 64.5; 64.4; 60.7; 59.6; 55.6; 46.1; 24.0

Example 17. 2 - (((3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1,1'-biphenyI] -4-yl) methyl) amino ) -2- (hydroxymethyl) propane-1,3-diol

The procedure described in Example 9 was used to prepare compound 17.

(3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1,1'-biphenyl] -4-yl) methanol (0.15 g, 0.39 mmol, 1.0 eq.), SOCI2 (0.14 mL, 1.95 mmol, 5.0 eq.), anhydrous DMF (0.04 mL), tris (hydroxymethyl) aminomethane (0.09 mL, 1.0 eq.) 95 mmol, 2.0 eq.), DIPEA (0.09 mL, 0.78 mmol, 2.0 eq.). The product 17 was obtained in a yield of 78% (0.15 g). Column chromatography: S1O2, chloroform / 7M NH3 in methanol, 20/1, v / v.

¹ H NMR (600 MHz, CDCl 3) δ [ppm]: 7.23 (d, J = 7.8 Hz, 1H), 7.21-7.17 (m, 3H), 6.90 (m, 3H ), 6.85-6.82 (m, 2H), 4.20 (s, 4H), 3.78 (s, 3H), 3.68 (s, 2H), 3.52 (s, 6H) , 3.13 (Wide, 3H) ¹³ C NMR (151 MHz, CDCl 3) δ [ppm]: 156.9; 143.2; 143.0; 141.3; 141.1; 140.9; 133.3; 130.9; 130.6; 130.1; 129.8; 127.1; 126.4; 122.8; 122.0; 118.6; 116.9; 112.2; 64.5; 64.4; 62.9; 60.4; 55.6; 40.9

Example 18 2 - (((2'-chloro-3 '- (benzo-1,4-dioxan-6-yl) -3-methoxy- [1,1'-biphenyl] -4-yl) methyl) amino ) -2- (hydroxymethyl) propane-1,3-diol

Intermediate 18.1 1 - ((3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1,1'-biphenyl] -4-yl) methyl) piperidine- Ethyl 4-carboxylate

The procedure described in Example 9 was followed to prepare compound 18.1.

(2'-chloro-3 '- (benzo-1,4-dioxan-6-yl) -3-methoxy- [1,1'-biphenyl] -4-yl) methanol (0.28 g, 0.74 mmol, 1.0 eq.), SOCI2 (0.27 mL, 3.71 mmol, 5.0 eq.), anhydrous DMF (0.06 mL), ethyl piperidine-4-carboxylate (0.34 mL, 2 , 22 mmol, 3.0 eq.), DIPEA (0.25 mL, 1.48 mmol, 2.0 eq.). Intermediate 18.1 was obtained in a yield of 7% (0.26 g). Column chromatography: S1O2, hexane / acetate, 2/1, v / v.

PL241 191 Β1 ¹ Η NMR (600 MHz, CDCl ₃ ) δ [ppm]: 7.43 (d, J = 7.1 Hz, 1H), 7.34-7.28 (m, 3H), 7.01 -6.99 (m, 2H), 6.97-6.91 (m, 3H), 4.31 (s, 4H), 4.13 (q, J = 7.1 Hz, 2H), 3. 84 (s, 2H), 3.59 (s, 2H), 3.00-2.93 (m, 2H), 2.34-2.26 (m, 1H), 2.18-2.10 ( m, 2H), 1.94-1.77 (m, 4H), 1.25 (t, J = 7.1 Hz, 3H) ¹³ C NMR (151 MHz, CDCl 2) δ [ppm]: 175.5 ; 157.3; 143.3; 143.1; 141.7; 141.2; 133.5; 131.1; 130.5; 130.3; 129.9; 126.4; 122.9; 121.7; 118.7; 117.0; 112.2; 64.6; 64.5; 60.4; 56.3; 55.7; 53.3; 41.4; 28.5; 14.4

End product 18.1 - ((3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1,1-biphenyl] -4-yl) methyl) piperidine-4- carbohydrazide

1 - ((3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1,1'-biphenyl] -4-yl) methyl) piperidine was placed in a round bottom flask Ethyl-4-carboxylate (0.24 g, 0.5 mmol, 1.0 eq.), Hydrazine monohydrate (0.94 ml, 20.0 mmol, 40.0 eq.) And methanol (10 ml). The contents of the flask were refluxed for 3 days with a magnetic stirrer. After this time, the solvent was evaporated. The product was purified using column chromatography (S1O2, methanol) to afford intermediate 18 in 28% yield (0.06 g).

¹ H NMR (600 MHz, DMSO-ds) δ [ppm]: 8.94 (s, 2H), 7.43 (t, J = 7.5 Hz, 1H), 7.39-7.36 (m , 2H), 7.34 (dd, J = 7.4; 1.9 Hz, 1H), 7.02 (d, J = 1.5 Hz, 1H), 6.99 (dd, J = 7, 7; 1.6 Hz, 1H), 6.94-6.92 (m, 2H), 6.89 (dd, J = 8.3; 2.0 Hz, 1H), 4.28 (s, 4H ), 3.80 (s, 3H), 3.47 (s, 2H), 2.90-2.85 (m, 2H), 2.08-2.02 (m, 1H), 1.98- 1.93 (m, 2H), 1.67-1.58 (m, 4H) ¹³ C NMR (151 MHz, DMSO-ds) δ [ppm]: 174.0; 156.8; 143.1; 142.9; 141.1; 140.5; 139.1; 132.5; 130.4; 130.3; 129.9; 129.1; 127.0; 125.6; 122.4; 121.2; 118.0; 116.7; 112.0; 64.1; 64.1; 55.6; 55.5; 53.0; 28.6; 20.5

Example 19. W- (2 - (((3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-ethoxy- [1,1-biphenyl] -4-yl) methyl) ) amino) ethyl) acetamide

The procedure described in Example 9 was used to prepare compound 19.

(3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-ethoxy- [1,1'-biphenyl] -4-yl) methanol (0.16 g, 0.27 mmol, 1.0 eq.), SOCI2 (0.98 ml, 1.35 mmol, 5.0 eq.), anhydrous DMF (0.03 ml), N - (2-aminoethyl) acetamide (0.05 mL, 0.54 mmol, 2.0 eq.), DIPEA (0.09 mL, 0.35 mmol, 2.0 eq.). The product 19 was obtained in a yield of 49% (0.09 g). Column chromatography: S1O2, chloroform / 7M NH3 in methanol, 20/1, v / v.

¹ H NMR (600 MHz, CDCl 3) δ [ppm]: 7.25-7.10 (m, 4H), 6.92 (d, J = 1.7 Hz, 1H), 6.91-6.89 (m, 2H), 6.87-6.84 (m, 2H), 6.14 (Wide, 1H), 4.22 (s, 4H), 4.02 (q, J = 7.0Hz, 2H), 3.75 (s, 2H), 3.29 (m, 2H), 2.70 (t, J = 5.9 Hz, 2H), 1.90 (s, 3H), 1.84 ( wide, 1H), 1.35 (t, J = 7.0 Hz, 3H).

¹³ CNMR (151 MHz, CDCl 3) δ [ppm]: 170.2; 156.4; 143.2; 143.0; 141.4; 141.1; 140.4; 133.3; 131.0; 130.5; 130.1; 129.5; 127.3; 126.4; 122.8; 121.5; 118.6; 116.9; 112.9; 64.5; 64.4; 63.6; 548.8; 47.8; 39.1; 23.4; 15.0

Example 20. W- (2 - (((3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-cyanomethoxy- [1,1'-biphenyl] -4-yl) methyl) amino) ethyl) acetamide

The procedure described in Example 9 was used to prepare compound 20.

2 - ((3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro -) - 4- (hydroxymethyl) - [1,1'-biphenyl] -3-yl) oxy) acetonitrile (0.22 g, 0.54 mmol, 1.0 eq.), SOCI2 (0.12 mL, 1.62 mmol, 3.0 eq.), Anhydrous DMF (0.04 mL), N- (2 -aminoethyl) acetamide (0.21 mL, 2.24 mmol, 4.0 eq.), DIPEA (0.21 mL, 2.24 mmol, 4.0 eq.). Product 20

PL241 191 Β1 was obtained with the efficiency of 9.8% (0.03 g). Column chromatography: S1O2, chloroform / 7M NH3 in methanol, 20/1, v / v.

¹ H NMR (600 MHz, CDCl 3) δ [ppm]: 7.37 (d, 7.7 Hz, 1H), 7.35-7.31 (m, 2H), 7.28 (dd, J = 6 , 7; 2.7 Hz, 1H), 7.15 (dd, J = 7.6; 1.5 Hz, 1H), 7.05 (d, J = 1.4 Hz, 1H), 6.99 -6.98 (m, 1H), 6.95-6.91 (m, 2H), 6.09 (s, 1H), 4.86 (s, 2H), 4.30 (s, 4H), 3.85 (s, 2H), 3.38 (q, J = 5.5 Hz, 2H), 2.81 (t, J = 5.6 Hz, 2H), 1.99 (s, 3H) ¹³ C NMR (151 MHz, CDCl 3) δ [ppm]: 170.3; 154.1; 143.3; 143.0; 141.2; 140.8; 140.5; 133.1; 130.9; 130.1; 128.5; 126.5; 124.3; 122.8; 118.5; 116.9; 115.1; 113.6; 64.5; 64.4; 53.9; 48.1; 48.0; 39.1; 23.4

Example 21. / V- (2 - (((3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3- (cyclobutylmethoxy) - [1,1 ^, -biphenyl] -4) -yl) methyl) amino) ethyl) acetamide.

The procedure described in Example 7 was used to prepare compound 21.

(3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3- (cyclobutylmethoxy) - [1, T-biphenyl] -4-yl) methanol (0.36 g, 0.15 g, 83 mmol, 1.0 eq.), SOCl2 (0.30 ml, 4.16 mmol, 5.0 eq.), Anhydrous DMF (0.05 ml), N - (2-aminoethyl) acetamide (0.1 23 mL, 2.49 mmol, 3.0 eq.), DIPEA (0.29 mL, 1.66 mmol, 2.0 eq.). The product 21 was obtained in a yield of 30% (0.13 g). Column chromatography: S1O2, chloroform / 7M NH3 in methanol, 20/1, v / v.

^1H NMR (600 MHz, DMSO-d6) δ [ppm]: 7.82 (t, J = 5.5 Hz, 1H), 7.43 (t, J = 7.6 Hz, 1H), 7, 38-7.36 (m, 2H), 7.35 (dd, J = 7.5; 1.8 Hz, 1H), 7.00 (d, J = 1.5 Hz, 1H), 6.97 (dd, J = 7.6, 1.6 Hz, 1H), 6.95-6.93 (m, 2H), 6.90 (dd, J = 8.3, 2.1 Hz, 1H), 4.29 (s, 4H), 3.99 (d, J = 6.3 Hz, 2H), 3.72 (s, 2H), 3.15 (q, J = 6.3 Hz, 2H), 2.78-2.71 (m, 1H), 2.58 (t, J = 6.5 Hz, 2H), 2.11-2.03 (m, 2H), 1.94-1.85 ( m, 4H), 1.79 (s, 3H) ¹³ C NMR (151 MHz, DMSO-d6) δ [ppm]: 169.1; 156.1; 143.1; 142.8; 141.1; 140.4; 139.0; 132.5; 130.4; 130.3; 129.9; 128.5; 128.5; 126.9; 122.4; 121.0; 118.0; 116.7; 112.7; 79.2; 71.3; 64.1; 64.1; 48.5; 47.3; 34.1; 24.2; 22.6; 18.1

Biological activity

NMR spectra measurements

NMR spectra for the following compounds were obtained on a Bruker Avance 600 MHz spectrometer at 300 K. Upon addition to the protein, all compounds tested dimerized human PD-L1 (0.18 mM) causing the peaks in the aliphatic part of the one-dimensional spectrum to disappear compared to the protein spectrum itself. An example of the human PD-L1 reference and d and me spectra after addition of compounds is shown in Fig. 1, which shows a comparison of ¹ H NMR spectra of the reference human PD-L1 protein (blue) after a 1: 5 addition of compound: 12 ( red), 17 (yellow), 6 (green), 20 (purple).

Determination of affinity of compounds by HTRF method

The biological activity of the compounds expressed in the form of IC50 - (half of the maximum inhibitory concentration) was determined by the Homogenous Time Resolved Fluorescence (HTRF) method. Determination was carried out with the use of a putty from the Cisbio company in the 20 μί format, using the protocol from the manufacturer. Compounds in the concentration range allowing determination of biological activity from the inflection curve were added to the hPD-1 (50 nM) and hPD-L1 (5 nM) protein complex with antibodies and allowed to incubate for 2 hours at room temperature before measuring Forster resonant energy transfer (TR-FRET) on a Tecan Spark 20M instrument. The results were normalized to positive (complex without inhibitor) and negative (no hPD-1) controls, averaged from three independent measurements and fitted with a Hill curve. Results shown in Table 1 at IC50 ranges> 100 nM (+), between 100 nM and 10 nM (++) and less than 100 nM (+++)

PL241 191 Β1

Name Biological activity Name Biological activity 1 + 12 +++ 2 + 13 +++ 6 + 14 +++ 7 ++ 15 +++ 9 ++ 16 ++ 11 ++ 17 +++

Results and discussion

Modulating the immune response by inhibiting the checkpoint receptors with antibodies is an effective method of fighting cancer known as immuno-oncology. The use of small molecule inhibitors instead of the known proteins is a beneficial change in cancer therapies due to the frequent and numerous adverse immunological effects caused by the use of antibodies.

In the light of the results of the studies described above, the compounds according to the invention constitute a new, attractive group of active inhibitors that bind directly to PD-L1, which was confirmed by the presented NMR studies, in which the compound dimerizes the protein, and the results of the HTRF tests performed. The observed dependence of structure on activity allows for additional optimization of the emerged structures.

ABBREVIATIONS:

EtOAc, ethyl acetate;

AcOH, acetic acid;

CHCl3, chloroform;

DMSO, dimethylsulfoxide

DCM, dichloromethane;

DMF, dimethylformamide;

HTRF, Homogenous Time Resolved Fluorescence

DIPEA, diisopropylethylamine;

MeOH, methanol;

ACN, acetone itrile;

MgSO4, magnesium sulfate;

NMR, Nuclear Magnetic Resonance Spectroscopy;

PD-1, Programmed Death 1;

PD-L1, Programmed Death Ligand;

THF, tetrahydrofuran;

EtO 2, diethyl ether

TLC, thin layer chromatography;

TMS, tetramethylsilane;

PL241 191 Β1

LITERATURE

1. Hellmann, M, D, et al, Nivolumab plus Ipilimumab in Advanced Non-Small-Cell Lung Cancer, N, Engl, J, Med, 381,2020-2031 (2019),

2. Hellmann, M, D, et al, Nivolumab plus ipilimumab in lung cancer with a high tumor mutational burden, N, Engl, J, Med, (2018) doi: 10,1056 / NEJMoa1801946,

3. Wolchok, J, D, et al, Nivolumab plus Ipilimumab in advanced melanoma, N, Engl, J, Med, (2013) doi: 10.1056 / NEJMoa1302369,

4. Pardoll, D, M, The blockade of immune checkpoints in cancer immunotherapy, Nat, Rev, Cancer 12, 252-264 (2012),

5. Tumeh, P, C, et al, PD-1 blockade induces responses by inhibiting adaptive immune resistance, Natura 515, 568-71 (2014),

6. Musielak, B, et al, CA-170 - A Potent Small-Molecule PD-L1 Inhibitor or Not? Molecules 24, 2804 (2019),

7. Adams, J, L, Smothers, J, Srinivasan, R, & Hoos, A, Big opportunities for smali molecules in immuno-oncology, Nat, Rev, Drug Discov, 14, 603-622 (2015),

8. Guzik, K, et al, Development of the Inhibitors That Target the PD-1 / PD-L1 Interaction-A Brief Look at Progress on Smali Molecules, Peptides and Macrocycles, Molecules 24, 2071 (2019.

Claims (5)

Patent claims 1. A compound of general formula (I): (1) where Ri is -H, -OCH2CN, an alkoxy group consisting of 1 to 6 carbon atoms, preferably containing alkane and methylcycloalkane moieties, an N, N-dialkyl group, diaryl or alkylarylcarbamate group, R2 is a substituent selected from the following structures: PL 241 191 B1 or a pharmaceutically acceptable salt thereof. 2. A compound according to claim A process as claimed in claim 1, which is selected from the group consisting of: (3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro- [1,1'-biphenyl] -4-yl) methanol, (3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1,1'-biphenyl] -4-yl) methanol, (2'-chloro-3' - (benzo-1,4-dioxan-6-yl) -3-methoxy- [1,1'-biphenyl] -4-yl) methanol, (3 '- (benzo-1,4-dioxan-6-yl) ) -2'-chloro-3-ethoxy- [1,1'-biphenyl] -4-yl) methanol, 2 - ((3 '- (benzo-1,4-dioxan-6-yl) -2'- chloro -) - 4- (hydroxymethyl) - [1,1'-biphenyl] -3-yl) oxy) - acetonitrile, (3 '- (benzo-1,4-dioxan-6-yl) -2'- chloro-3- (cyclobutyl) methoxy- [1,1'-biphenyl] -4-yl) methanol, 3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-4- (hydroxymethyl ) - [1,1'-biphenyl] -3-yl) dimethylcarbamate, 1 - ((3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1 , 1'-biphenyl] -4-yl) methyl) piperazine, 1-carboxamide 4 - ((3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1 , 1'-biphenyl] -4-yl) methyl) piperazine, 1- (4 - ((3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1,1'-biphenyl] -4-yl) methyl) piperazin- 1-yl) ethan-1-one, 2-amino-1- (4 - ((3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1, 1'-biphenyl] -4-yl) methyl) piperazin-1-yl) ethan-1-one, 1 - ((3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro 3-methoxy- [1,1'-biphenyl] -4-yl) methyl) -4- (methylsulfonyl) piperazine, N- (2 - (((3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1,1'-biphenyl] -4-yl) methyl) amino ) ethyl) acetamide, 2-amino-N- (2 - ((3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1,1'-biphenyl] -4-yl) methyl) amino) ethyl) acetamide, N- (2 - (((3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1,1'-biphenyl] -4-yl) methyl) amino ) ethyl) methanesulfonamide, 1- (2 - (((3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1,1'-biphenyl] -4-) yl) methyl) amino) ethyl) urea, 2 - (((3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1,1'-biphenyl] - 4-yl) methyl) amino) propane-1,3-diol, 2 - (((3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-methoxy- [1,1' -biphenyl] -4-yl) methyl) amino) -2- (hydroxymethyl) propane-1,3-diol, 2 - (((2'-chloro-3 '- (benzo-1,4-dioxane-6- yl) -3-methoxy- [1,1'-biphenyl] -4-yl) methyl) amino) -2- (hydroxymethyl) propane-1,3-diol, N- (2 - (((3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-ethoxy- [1,1'-biphenyl] -4-yl) methyl) amino ) ethyl) acetamide, N- (2 - (((3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3-cyanomethoxy- [1,1'-biphenyl] -4-yl) methyl) amino ) ethyl) acetamide and N- (2 - (((3 '- (benzo-1,4-dioxan-6-yl) -2'-chloro-3- (cyclobutylmethoxy) - [1,1'-biphenyl] -4-yl) methyl) ) amino) ethyl) acetamide. 3. A compound as defined in claim 1-2 for use as an inhibitor of the PD-I / PD-L1 protein pathway, especially in mammalian cells, preferably tumor cells. 4. A compound as defined in claim 1-2 for use in pharmacy, especially in the treatment or prophylaxis of neoplastic diseases. 5. A pharmaceutical composition containing a therapeutically effective dose of a compound according to claim 1 1-2.