Danoprevir

Danoprevir is a NS3/4A protease inhibitor for hepatitis C virus (HCV) with IC50 of 0.2-3.5 nM. The inhibition effect on HCV genotypes 1A/1B/4/5/6 is approximately 10-fold higher than 2B/3A.

Danoprevir has been used in trials studying the treatment of Hepatitis C, Chronic.|Danoprevir is an orally bioavailable, peptidomimetic inhibitor of hepatitis C virus (HCV) NS3/4A protease, with antiviral activity against HCV and potential antiviral activity against SARS-CoV-2. Upon oral administration, danoprevir binds to and blocks the activity of HCV NS3/4A protease. This prevents the cleavage and processing of HCV viral proteins leading to the inhibition of HCV replication. Danoprevir may also bind to and block of the activity of SARS-CoV-2 protease. This prevents the cleavage and processing of SARS-CoV-2 viral proteins leading to the inhibition of SARS-CoV-2 replication. NS3/4A, a chymotrypsin-like serine protease, is responsible for cleavage at four sites of the HCV polyprotein to form the viral proteins required for HCV replication. It plays a key role in the HCV viral replication process. HCV infection is associated with the development of hepatocellular carcinoma (HCC). A chymotrypsin-like protease is responsible for cleavage of the SARS-CoV-2 viral polyprotein to form the RNA replicase-transcriptase complex, which plays a key role in the SARS-CoV-2 viral transcription and replication process.

To a suspension of 30.0 g (0.043 mol) of carboxylic acid (product of example 20 with an assay of 90.2%(m/m)) and 14.0 g of sodium carbonate in 225 g of tetrahydrofuran was added at 45C within 30 minutes 7.60 g (0.074 mol) of acetic acid anhydride and the resulting mixture was stirred at 45C for 8 hours. To the resulting suspension was then added 30.2 g (0.17mol) of potassium carbonate and 8.0 g (0.065 mol) of cyclopropyl sulfonamide. The mixture was heated to 62C and stirred at this temperature for 17 hours. The mixture was concentrated to a residual volume of 200 ml and then treated with 200 g of water. The biphasic mixture was stirred for 15 minutes and the layers were then allowed to separate. The lower aqueous phase was removed. The organic phase was diluted with 90 g of ethyl acetate and washed with 3% sulfuric acid (1x140 g) and water (3x130 g). The organic layer was concentrated to dryness and then diluted with 400 ml of ethyl acetate. Residual amounts of water were removed by a continuous azeotropic distillation with ethyl acetate. The mixture was then treated at 100C with 20 ml of methanol, followed by 10.0 g of sodium methylate (30% in methanol). From the resulting Preparation of Sodium ((2R, 6S, 13aS, 14aR, 16aS, Z)-6-(tert-butoxycarbonylamino)-2-(4-fluoroisoiindoline-2-carbonyloxy)-5, 16-dioxo-1, 2, 3, 5, 6, 7, 8, 9, 10, 11, 13a, 15, 16a-hexadecahydrocyclopropa[e]pyrrolo[1, 2-a][1, 4]diazacyclopentadecine-14a-carbonyl) (cyclopropylsulfonyl)amide (HCV protease inhibitor; compound VIII)To a suspension of 30.0 g (0.043 mol) of XXb from with an assay of 90.2%(m/m)) and 14.0 g of sodium carbonate in 225 g of tetrahydrofuran was added at 45 C. within 30 minutes 7.60 g (0.074 mol) of acetic acid anhydride and the resulting mixture was stirred at 45 C. for 8 hours. To the resulting suspension was then added 30.2 g (0.17 mol) of potassium carbonate and 8.0 g (0.065 mol) of cyclopropyl sulfonamide. The mixture was heated to 62 C. and stirred at this temperature for 17 hours. The mixture was concentrated to a residual volume of 200 mL and then treated with 200 g of water. The biphasic mixture was stirred for 15 minutes and the layers were then allowed to separate. The lower aqueous phase was removed. The organic phase was diluted with 90 g of ethyl acetate and washed with 3% sulfuric acid (1×140 g) and water (3×130 g). The organic layer was concentrated to dryness and then redissolved in 400 mL of ethyl acetate. Residual amounts of water were removed by a continuous azeotropic distillation with ethyl acetate. The mixture was then treated at 10 C. with 20 mL of methanol, followed by 10.0 g of sodium methylate (30% in methanol). From the resulting mixture approx. 300 mL of ethyl acetate/methanol were then distilled off. The mixture was then treated at 34 C. within one hour with 300 mL of ethyl acetate and 5 g of water. The resulting mixture was allowed to cool to ambient temperature within 4 hours. The crystals were filtered off, washed with 80 mL of ethyl acetate and dried at 80 C./<30 mbar for 20 hours to afford 30.4 g (87% corrected yield) of the title compound as white crystals with an assay of 92.7%(m/m).MS: 732.28 (M++H), 676.23, 632.25.1H-NMR (400 MHz, DMSO-d6): 7.89-7.80 (m, 1H), 7.39-7.31 (m, 1H), 7.21-7.06 (m, 2H), 6.97-6.90 (m, 1H), 5.49-4.41 (m, 1H), 5.31-5.21 (m, 2H), 4.66 (s, br, 4H), 4.45-4.35 (m, 1H), 4.19-4.08(m, 2H), 3.91-3.81 (m, IH), 2.68-2.58(m, 1H), 2.30-2.14 (m, 3H), 2.0-1.2 (m, 12H), 1.17and 1.14 (2s, 9H), 0.78-0.69 (m, 2H), 0.62-0.53 (m, 2H).To a suspension of 30.0 g (0.043 mol) of carboxylic acid (product of example 20 with an assay of 90.2%(m/m)) and 14.0 g of sodium carbonate in 225 g of tetrahydrofuran was added at 45C within 30 minutes 7.60 g (0.074 mol) of acetic acid anhydride and the resulting mixture was stirred at 45C for 8 hours. To the resulting suspension was then added 30.2 g (0.17mol) of potassium carbonate and 8.0 g (0.065 mol) of cyclopropyl sulfonamide. The mixture was heated to 62C and stirred at this temperature for 17 hours. The mixture was concentrated to a residual volume of 200 ml and then treated with 200 g of water. The biphasic mixture was stirred for 15 minutes and the layers were then allowed to separate. The lower aqueous phase was removed. The organic phase was diluted with 90 g of ethyl acetate and washed with 3% sulfuric acid (1x140 g) and water (3x130 g). The organic layer was concentrated to dryness and then diluted with 400 ml of ethyl acetate. Residual amounts of water were removed by a continuous azeotropic distillation with ethyl acetate. The mixture was then treated at 100C with 20 ml of methanol, followed by 10.0 g of sodium methylate (30% in methanol). From the resulting To a suspension of 30.0 g (0.043 mol) of carboxylic acid (product of with an assay of 90.2%(m/m)) and 14.0 g of sodium carbonate in 225 g of tetrahydrofuran was added at 45 C. within 30 minutes 7.60 g (0.074 mol) of acetic acid anhydride and the resulting mixture was stirred at 45 C. for 8 hours. To the resulting suspension was then added 30.2 g (0.17 mol) of potassium carbonate and 8.0 g (0.065 mol) of cyclopropylsulfonamide. The mixture was heated to 62 C. and stirred at this temperature for 17 hours. The mixture was concentrated to a residual volume of 200 mL and then treated with 200 g of water. The biphasic mixture was stirred for 15 minutes and the layers were then allowed to separate. The lower aqueous phase was removed. The organic phase was diluted with 90 g of ethyl acetate and washed with 3% sulfuric acid (1×140 g) and water (3×130 g). The organic layer was concentrated to dryness and then diluted with 400 mL of ethyl acetate. Residual amounts of water were removed by a continuous azeotropic distillation with ethyl acetate. The mixture was then treated at 10 C. with 20 mL of methanol, followed by 10.0 g of sodium methylate (30% in methanol). From the resulting mixture approx. 300 mL of ethyl acetate/methanol were then distilled off. The mixture was then treated at 34 C. within one hour with 300 mL of ethyl acetate and 5 g of water. The resulting mixture was allowed to cool to ambient temperature within 4 hours. The crystals were filtered off, washed with 80 mL of ethyl acetate and dried at 80 C./<30 mbar for 20 hours to afford 30.4 g (87% corrected yield) of the title compound as white crystals with an assay of 92.7%(m/m):MS: 732.28 (M++H), 676.23, 632.25; 1H-NMR (400 MHz, DMSO-d6): 7.89-7.80 (m, 1H), 7.39-7.31 (m, 1H), 7.21-7.06 (m, 2H), 6.97-6.90 (m, 1H), 5.49-4.41 (m, 1H), 5.31-5.21 (m, 2H), 4.66 (s, br, 4H), 4.45-4.35 (m, 1H), 4.19-4.08(m, 2H), 3.91-3.81 (m, 1H), 2.68-2.58(m, 1H), 2.30-2.14 (m, 3H), 2.0-1.2 (m, 12H), 1.17 and 1.14 (2s, 9H), 0.78-0.69 (m, 2H), 0.62-0.53 (m, 2H).Preparation of Sodium ((2R, 6S, 13aS, 14aR, 16aS, Z)-6-(tert-butoxycarbonylamino)-2-(4-fluoroisoiindoline-2-carbonyloxy)-5, 16-dioxo-1, 2, 3, 5, 6, 7, 8, 9, 10, 11, 13a, 15, 16a-hexadecahydrocyclopropa[e]pyrrolo[1, 2-a][1, 4]diazacyclopentadecine-14a-carbonyl) (cyclopropylsulfonyl)amide (HCV protease inhibitor; compound VIII)To a suspension of 30.0 g (0.043 mol) of XXb from with an assay of 90.2%(m/m)) and 14.0 g of sodium carbonate in 225 g of tetrahydrofuran was added at 45 C. within 30 minutes 7.60 g (0.074 mol) of acetic acid anhydride and the resulting mixture was stirred at 45 C. for 8 hours. To the resulting suspension was then added 30.2 g (0.17 mol) of potassium carbonate and 8.0 g (0.065 mol) of cyclopropyl sulfonamide. The mixture was heated to 62 C. and stirred at this temperature for 17 hours. The mixture was concentrated to a residual volume of 200 mL and then treated with 200 g of water. The biphasic mixture was stirred for 15 minutes and the layers were then allowed to separate. The lower aqueous phase was removed. The organic phase was diluted with 90 g of ethyl acetate and washed with 3% sulfuric acid (1×140 g) and water (3×130 g). The organic layer was concentrated to dryness and then redissolved in 400 mL of ethyl acetate. Residual amounts of water were removed by a continuous azeotropic distillation with ethyl acetate. The mixture was then treated at 10 C. with 20 mL of methanol, followed by 10.0 g of sodium methylate (30% in methanol). From the resulting mixture approx. 300 mL of ethyl acetate/methanol were then distilled off. The mixture was then treated at 34 C. within one hour with 300 mL of ethyl acetate and 5 g of water. The resulting mixture was allowed to cool to ambient temperature within 4 hours. The crystals were filtered off, washed with 80 mL of ethyl acetate and dried at 80 C./<30 mbar for 20 hours to afford 30.4 g (87% corrected yield) of the title compound as white crystals with an assay of 92.7%(m/m).MS: 732.28 (M++H), 676.23, 632.25.1H-NMR (400 MHz, DMSO-d6): 7.89-7.80 (m, 1H), 7.39-7.31 (m, 1H), 7.21-7.06 (m, 2H), 6.97-6.90 (m, 1H), 5.49-4.41 (m, 1H), 5.31-5.21 (m, 2H), 4.66 (s, br, 4H), 4.45-4.35 (m, 1H), 4.19-4.08(m, 2H), 3.91-3.81 (m, IH), 2.68-2.58(m, 1H), 2.30-2.14 (m, 3H), 2.0-1.2 (m, 12H), 1.17and 1.14 (2s, 9H), 0.78-0.69 (m, 2H), 0.62-0.53 (m, 2H).To a suspension of 30.0 g (0.043 mol) of carboxylic acid (product of with an assay of 90.2%(m/m)) and 14.0 g of sodium carbonate in 225 g of tetrahydrofuran was added at 45 C. within 30 minutes 7.60 g (0.074 mol) of acetic acid anhydride and the resulting mixture was stirred at 45 C. for 8 hours. To the resulting suspension was then added 30.2 g (0.17 mol) of potassium carbonate and 8.0 g (0.065 mol) of cyclopropylsulfonamide. The mixture was heated to 62 C. and stirred at this temperature for 17 hours. The mixture was concentrated to a residual volume of 200 mL and then treated with 200 g of water. The biphasic mixture was stirred for 15 minutes and the layers were then allowed to separate. The lower aqueous phase was removed. The organic phase was diluted with 90 g of ethyl acetate and washed with 3% sulfuric acid (1×140 g) and water (3×130 g). The organic layer was concentrated to dryness and then diluted with 400 mL of ethyl acetate. Residual amounts of water were removed by a continuous azeotropic distillation with ethyl acetate. The mixture was then treated at 10 C. with 20 mL of methanol, followed by 10.0 g of sodium methylate (30% in methanol). From the resulting mixture approx. 300 mL of ethyl acetate/methanol were then distilled off. The mixture was then treated at 34 C. within one hour with 300 mL of ethyl acetate and 5 g of water. The resulting mixture was allowed to cool to ambient temperature within 4 hours. The crystals were filtered off, washed with 80 mL of ethyl acetate and dried at 80 C./<30 mbar for 20 hours to afford 30.4 g (87% corrected yield) of the title compound as white crystals with an assay of 92.7%(m/m):MS: 732.28 (M++H), 676.23, 632.25; 1H-NMR (400 MHz, DMSO-d6): 7.89-7.80 (m, 1H), 7.39-7.31 (m, 1H), 7.21-7.06 (m, 2H), 6.97-6.90 (m, 1H), 5.49-4.41 (m, 1H), 5.31-5.21 (m, 2H), 4.66 (s, br, 4H), 4.45-4.35 (m, 1H), 4.19-4.08(m, 2H), 3.91-3.81 (m, 1H), 2.68-2.58(m, 1H), 2.30-2.14 (m, 3H), 2.0-1.2 (m, 12H), 1.17 and 1.14 (2s, 9H), 0.78-0.69 (m, 2H), 0.62-0.53 (m, 2H).The starting material (AR00334191, Example 3-55, 10 mg, 13. 7 umol) was dissolved in 1 mL of 50 % TFA (DCM) and stirred at RT for 1 h. The reaction mixture was then concentrated to dryness, taken up in acetonitrile and concentrated again. Repeat the above process once more to remove any excess TFA. The resulting solid residue was then dissolved in DCE (137 I1L), cooled to 0 C in an ice bath, followed by addition of the amino acid, cyclohexyl-[(pyrazine-2-carbonyl)-amino]-acetic acid (1.05 equiv), HATU (10 mg) and DIEA (4 drps). The mixture was let slowly warm up to RT and stir for overnight. For work-up, the reaction mixture was directly loaded onto a C-18 column and purified with reverse-phase column chromatography, giving the target compound as a white solid. MS (APCI-): m/z 876.1 (M-1).The starting material (AR00334191, Example 3-55, 10 mg, 13.7 pmol) was dissolved in 1 mL of 50 % TFA (DCM) and stirred at RT for 1 h. The reaction mixture was then concentrated to dryness, taken up in acetonitrile and concentrated again. Repeat the above process once more to remove any excess TFA. The resulting solid residue was then dissolved in DCE (137 L), followed by addition of the acid chloride, (4-Methoxy-phenyl)-acetyl chloride (2 drps) and DIEA (4 drps). The mixture was stirred ar RT for overnight. After completion, the reaction was directly loaded onto a C-18 column and purified with reverse- phase column chromatography. The compound was further purified on normal phase silica gel chromatography (eluent = 40 % EtOAc/hexanes with 1 % formic acid) to give the target compound as a white solid. 1H NMR (500 MHz, CD30D) 8 7.33 (p, 1H), 7.15 (d, 1H), 7.05- 6.92 (m, 3H), 6.65 (dd, 2H), 5.68 (q, 1H), 5.40 (br s, 1H), 5.09 (t, 1H), 4.78-4. 46 (m, 7H), 4.43-4. 24 (m, 2H), 3. 89-3. 80 (m, 1H), 3.68 (d, 3H), 3.21 (d, 1H), 2.69-2. 57 (m, 1H), 2.52 - 2. 30 (m, 5H), 2.06-0. 80 (m, 15H); MS (APCI-): m/z 778.3 (M-1).

Molecular Weight:731.8
XLogP3:3.3
Hydrogen Bond Donor Count:3
Hydrogen Bond Acceptor Count:10
Rotatable Bond Count:8
Exact Mass:731.30002740
Monoisotopic Mass:731.30002740
Topological Polar Surface Area:189
Heavy Atom Count:51
Complexity:1530
Defined Atom Stereocenter Count:5
Defined Bond Stereocenter Count:1
Covalently-Bonded Unit Count:1
Compound Is Canonicalized:Yes