Tizoxanide carbamate and pharmaceutical use thereof

Abstract

The invention relates to a tizoxanide carbamate compound of Formula I and a pharmaceutically acceptable salt, a hydrate or a solvate thereof, and their pharmaceutical use. ##STR00001##

Claims

1. A compound selected from the following structures: ##STR00011## or a pharmaceutically acceptable salt, a hydrate or a solvate thereof.

2. A pharmaceutical composition, comprising the compound according to claim 1, a pharmaceutically acceptable salt, a hydrate or a solvate thereof.

3. The pharmaceutical composition according to claim 2, further comprising a pharmaceutically acceptable carrier or excipient.

4. The pharmaceutical composition according to claim 3, said pharmaceutical composition is in the form of a solid preparation, an injection, an external preparation, a spray, a liquid preparation or a compound preparation.

5. A method for treating a disease in a mammal in need thereof, comprising administering to the mammal in need thereof a therapeutically effective amount of the compound according to claim 1, or a pharmaceutically acceptable salt, hydrate or solvate thereof, wherein the disease is selected from an infection by a parasite, hepatitis B, hepatitis C, influenza, a viral infectious disease caused by Rotavirus or Norovirus, and an infection caused by a bacterium.

6. A method for inhibiting biofilm formation caused by a bacterium in a mammal in need thereof, comprising administering to the mammal in need thereof a therapeutically effective amount of the compound according to claim 1 or a pharmaceutically acceptable salt, hydrate or solvate thereof.

7. The method according to claim 5, wherein parasite is protozoan, or helminth.

8. The method according to claim 5, wherein said viral infection disease caused by Rotavirus or Norovirus is viral enterogastritis caused by Rotavirus or Norovirus.

9. The method according to claim 5, wherein said infection caused by a bacterium is an infection caused by Clostridium difficile or Tubercle bacillus or Helicobacter pylori.

10. The method according to claim 8, wherein said Tubercle bacillus is drug-resistant Tubercle bacillus.

11. The method according to claim 5, wherein said parasite is selected from the group consisting of: Giardia, Amoeba, Cryptosporidium, Cyclospora, Trichomonad, Encephalitozoon intestinalis, Isospora belli, Blastocystis hominis, Balantidium coli, Ascarislumbricoides, Enterocytozoon bieneusi, Tapeworm, Diplacanthus nanus, Giardia lamblia, Leishmania, and Fasciola hepatica.

12. The method according to claim 11, wherein said parasite is Tapeworm and said Tapeworm is Taenia saginata, or Hymenolepis nana.

Description

DESCRIPTION OF THE DRAWINGS

(1) FIG. 1: The concentration-time curves of tizoxanide in plasma after oral administration of nitazoxanide and Compound 1, 2, 3, 4, 5, 6 in mice, respectively.

SPECIFIC MODES FOR CARRYING OUT THE INVENTION

(2) The embodiments of the invention are described by combining the following examples. However, a person skilled in the art understands that the following examples are only intended to describe the invention, and shall not be regarded as defining the scope of the invention. When the particular techniques or conditions are not indicated in Examples, the invention is carried out according to the techniques or conditions described in the prior art documents or according to the product instruction. The reagents or apparatuses, the manufacturers of which are not indicated, are the conventional products that are commercially available.

EXAMPLE 1

Preparation of 2-(5-nitrothiazol-2-yl-carbamoyl)phenylmorpholine-4-carboxylate (Compound 1)

(3) ##STR00005##

(4) In an ice bath, to a reaction bottle, a solution of solid triphosgene (594 mg, 1.85 mmol) in dichloroethane (20 mL) was added, and a solution of morpholine (0.436 g, 5.0 mmol) dissolved in pyridine (8 mL) was added slowly dropwise. The reaction was carried out in the ice bath for 4-5 h. N.sub.2 gas was introduced to remove the excessive triphosgene, the ice bath was removed, a solution of tizoxanide (1.325 g, 5.0 mmol) in THF (10 mL) was added at room temperature, and the reaction was carried out under stirring overnight. The product was extracted with ethyl acetate, the ethyl acetate phase was washed sequentially with water and saturated salt solution, the organic phase was dried with anhydrous sodium sulphate, the solvent was removed by distillation under reduced pressure, and the resultant residue was subjected to silica gel column chromatography, to obtain the product 1.23 g, with a yield of 65.08%, .sup.1H-NMR (CDCl.sub.3, 400 Hz) ppm: 10.66 (s, 1H), 8.19 (s, 1H), 7.93 (dd, J=1.56, 7.93 Hz, 1H), 7.66-7.62 (m, 1H), 7.42-7.39 (m, 1H), 7.25-7.26 (m, 1H), 3.76-3.81 (m, 6H), 3.60-3.61 (m, 2H). ESI-MS m/z: 379.3[MH].sup.+, 401.3[MNa].sup.+.

EXAMPLE 2

Preparation of 2-(5-nitrothiazol-2-yl-carbamoyl)phenyl-4-methylpiperazine-1-carboxylate (Compound 2)

(5) ##STR00006##

(6) In an ice bath, to a reaction bottle, a solution of solid triphosgene (594 mg, 1.85 mmol) in dichloroethane (20 mL) was added, and a solution of 1-methyl piperidine (0.501 g, 5.0 mmol) dissolved in pyridine (8 mL) was added slowly dropwise. The reaction was carried out in the ice bath for 4-5 h. N.sub.2 gas was introduced to remove the excessive triphosgene, the ice bath was removed, and a solution of tizoxanide (1.325 g, 5.0 mmol) in THF (10 mL) was added at room temperature. The reaction was carried out under stirring overnight. The product was extracted with ethyl acetate, the ethyl acetate phase was washed sequentially with water and saturated salt solution, the organic phase was dried with anhydrous sodium sulphate, the solvent was removed by distillation under reduced pressure, and the resultant residue was subjected to silica gel column chromatography, to obtain the product 0.96 g, with a yield of 48.98%. .sup.1H-NMR (CDCl.sub.3, 400 Hz) ppm: 8.25 (s, 1H) 7.92 (dd, J=1.68, 7.84 Hz, 1H), 7.65-7.60 (m, 1H), 7.41-7.37 (m, 1H), 7.24 (d, J=8.12 Hz, 2H), 3.83 (brs, 2H), 3.65 (brs, 2H), 2.62 (brs, 2H), 2.53 (brs, 2H), 2.41 (s, 3H). ESI-MS m/z: 392 [MH].sup.+, 414 [MNa].sup.+.

EXAMPLE 3

Preparation of 2-((5-nitrothiazol-2-yl)carbamoyl)phenyl-piperidine-4-carboxylate (Compound 3)

(7) ##STR00007##

(8) In an ice bath, to a reaction bottle, a solution of solid triphosgene (594 mg, 1.85 mmol) in dichloroethane (20 mL) was added, and a solution of piperidine (0.43 g, 5.0 mmol) dissolved in pyridine (8 mL) was added slowly dropwise. The reaction was carried out in the ice bath for 4-5 h. N.sub.2 gas was introduced to remove the excessive triphosgene, the ice bath was removed, and a solution of tizoxanide (1.325 g, 5.0 mmol) in THE (10 mL) was added at room temperature. The reaction was carried out under stirring overnight. The product was extracted with ethyl acetate, the ethyl acetate phase was washed sequentially with water and saturated salt solution, the organic phase was dried with anhydrous sodium sulphate, the solvent was removed by distillation under reduced pressure, and the resultant residue was subjected to silica gel column chromatography, to obtain the product 1.26 g, with a yield of 67.02%, .sup.1H-NMR (CDCl.sub.3, 400 Hz) ppm: 8.28 (s, 1H), 7.97 (dd, J=1.68, 7.84 Hz, 1H), 7.64-7.60 (m, 1H), 7.41-7.37 (m, 1H), 7.26-7.22 (m, 1H), 3.72 (br s, 2H), 3.55 (br s, 2H), 1.65 (br s, 6H). ESI-MS m/z: 377 [MH].sup.+, 399 [MNa].sup.+.

EXAMPLE 4

Preparation of 2-((5-nitrothiazol-2-yl)carbamoyl)phenyl-pyrrolidine-1-carboxylate (Compound 4)

(9) ##STR00008##

(10) In an ice bath, to a reaction bottle, a solution of solid triphosgene (594 mg, 1.8 mmol) in dichloroethane (20 mL) was added, and a solution of pyrrole (0.345 g, 5.0 mmol) dissolved in pyridine (8 mL) was added slowly dropwise. The reaction was carried out in the ice bath for 4-5 h. N.sub.2 gas was introduced to remove the excessive triphosgene, the ice bath was removed, and a solution of tizoxanide (1.325 g, 5.0 mmol) in THE (10 mL) was added at room temperature. The reaction was carried out under stirring overnight. The product was extracted with ethyl acetate, the ethyl acetate phase was washed sequentially with water and saturated salt solution, the organic phase was dried with anhydrous sodium sulphate, the solvent was removed by distillation under reduced pressure, and the resultant residue was subjected to silica gel column chromatography, to obtain the product 1.24 g, with a yield of 68.51%, .sup.1H-NMR(CDCl.sub.3, 400 Hz) ppm: 11.08 (s, 1H), 8.22 (s, 1H), 8.00 (dd, J=1.82, 7.70 Hz, 1H), 7.65-7.60 (m, 1H), 7.40-7.31 (m, 2H), 3.73 (t, J=6.72 Hz, 2H), 3.52 (t, J=6.72 Hz, 2H), 2.06-1.96 (m, 4H). ESI-MS m/z: 363[MH].sup.+, 385[MNa].sup.+.

EXAMPLE 5

Preparation of 2-(5-nitrothiazol-2-yl-carbamoyl)phenyl-4-(2-ethoxy-2-oxoethyl) piperidine-1-carboxylate (Compound 5)

(11) ##STR00009##

(12) In an ice bath, to a reaction bottle, a solution of triphosgene (594 mg, 2 mmol) in dichloroethane (10 mL), ethyl piperidine-4-carboxylate (786 mg, 500 mmol), and a solution of triethylamine (0.7 mL) in dichloroethane (5 mL) were added sequentially. After stirring at room temperature for 2 h, the reaction mixture was distilled under reduced pressure, and the resultant residue was dissolved in anhydrous THE (10 mL), and in an ice bath again, a solution of tizoxanide (1.325 g) in THF (10 mL), triethylamine (549 mg, 5.0 mmol), and 4-dimethylaminopyridine (DMAP) 100 mg were added. The mixture was stirred at room temperature overnight, and distilled water was then added to the reaction mixture. The resultant mixture was extracted with ethyl acetate, the ethyl acetate phase was washed sequentially with water and saturated salt solution, the organic phase was dried with anhydrous sodium sulphate, the solvent was removed by distillation under reduced pressure, and the resultant residue was subjected to silica gel column chromatography, to obtain the product 0.73 g, with a yield of 55.09%, .sup.1H-NMR (DMSO-d6,400 Hz) ppm: 13.57 (s, 1H), 8.68 (s, 1H), 7.77 (d, J=7.6 Hz, 1H), 7.67-7.63 (m, 1H), 7.43-7.39 (m, 1H), 7.31-7.33 (m, 1H), 4.06-4.11 (m, 3H), 3.81-3.84 (m, 1H), 3.10-3.17 (m, 1H), 2.87-2.93 (m, 1H), 1.66-1.88 (m, 3H), 1.36-1.39 (m, 1H), 1.19 (t, J=7.2 Hz, 3H). ESI-MS: m/z: 449.1[MH].sup.+, 471.1[MNa].sup.+.

EXAMPLE 6

Preparation of 2-(5-nitrothiazol-2-yl-carbamoyl)phenyl-4-(2-ethoxy-2-oxoethyl) piperidine-1-carboxylate (Compound 6)

(13) ##STR00010##

(14) To a suspension of tizoxanide (10 g, 39 mmol) in DMF (80 mL), triethylamine (7.97 g, 78 mmol) was added, and the mixture was stirred until tizoxanide was completely dissolved. To the resultant mixture, a solution of N,N-Dimethyl carbamoyl chloride (8.39 g, 78 mmol) in 20 mL DMF, and DMAP (200 mg) were added dropwise, and the mixture was stirred at room temperature overnight. The reaction solution was poured into 300 mL ice-water containing 7.5 mL concentrated hydrochloric acid. The insoluble substance was removed by filtration. The filtrate was extracted with dichloromethane 3 times, the dichloromethane phase was washed sequentially with water and saturated salt solution, and was dried with anhydrous sodium sulphate, the solvent was removed by distillation under reduced pressure, and the resultant residue was subjected to silica gel column chromatography, to obtain the product 6 g, with a yield of 45%. .sup.1H-NMR (DMSO-d6, 400 Hz) ppm: 13.53 (s, 1H), 8.70 (s, 1H), 7.76 (d, J=7.6 Hz, 1H), 7.62-7.67 (m, 1H), 7.37-7.41 (m, 1H), 7.31-7.33 (m, 1H), 3.03 (s, 3H), 2.83 (m, ESI-MS: m/z: 337.1[MH].sup.+.

EXAMPLE 7

Pharmacokinetic Evaluation and Result of Oral Administration of the Compounds in Mice

(15) ICR mice (SPS grade, male, 252 mg), were grouped randomly depending on body weight, 3 mice for each group. The subject compound nitazoxanide and Compound 1, 2, 3, 4, 5, 6 (Sample preparation: 0.03257 mmol of the subject compound was dissolved in 100 uL DMSO, with the addition of 10 mL 0.5% CMC-Na solution, to prepare a sample at 3.257 mol/mL, and the sample was intragastrically administered at a dose of 10 mL/kg (equivalent to a dose of 32.57 mol/kg tizoxanide) to each mice) were intragastically administered at a dose equivalent to 32.57 mol/kg tizoxanide, wherein nitazoxanide was used as positive control, and 0.1 ml blood was taken from veins of Fundus Oculi at 0.08, 0.25, 0.5, 1, 2, 4, 6, 8 and 12 h after administration, placed in a Heparin Sodium-containing centrifuge tube, and centrifuged at 3000 g for 20 min. The plasma was taken and stored in a 20 C. refrigerator for further detection.

(16) During the detection, 50 L plasma was taken, with the addition of an internal standard solution (5000 ng/ml, glipizide aqueous solution, 10 L), and the addition of acetonitrile (200 L), then mixed well. The mixture was vortexed for 3 min, and centrifuged for 10 min (at a relative centrifugal force (RCF) of 8500 g). The supernatant (50 L) was mixed with water (50 L), and 10 L was drawn for LC/MS/MS analysis, so as to detect the blood concentration of tizoxanide. The results are shown in Table 1, Table 2 and FIG. 1.

(17) TABLE-US-00001 TABLE 1 Blood concentrations (Mean SD, n = 3) of tizoxanide after oral administration of the compound nitazoxanide and Compound 1, 2, 3, 4, 5, 6 in mice, respectively concentration (ng/mL) time Compound Compound Compound Compound Compound Compound (h) NTZ 1 2 3 4 5 6 0.08 127.23 120.62 495.61 139.51 156.67 329.85 25.33 22.46 53.49 161.89 20.57 34.92 37.88 8.09 0.25 329.39 397.98 444.34 254.20 411.33 148.92 128.02 111.17 72.65 80.08 67.14 133.87 24.08 23.18 0.5 94.65 535.69 222.08 787.61 561.54 92.03 275.35 24.48 267.17 93.37 90.92 129.43 11.37 81.27 1 19.69 468.37 233.28 812.20 705.78 48.61 430.11 5.62 235.43 44.18 199.523 113.32 8.32 155.20 2 13.91 154.82 200.57 541.76 477.14 13.39 478.64 3.60 68.23 78.52 49.39 127.03 2.27 103.31 4 8.22 57.26 79.88 226.51 204.24 4.18 266.56 5.08 34.49 126.76 92.02 36.11 1.51 31.28 6 3.46 52.25 5.83 135.86 45.94 6.14 131.48 1.66 53.09 5.45 18.47 30.08 4.10 24.35 8 0.61 21.74 3.87 44.67 33.29 2.90 104.15 0.12 5.95 4.45 4.88 47.42 1.04 95.20 12 4.90 4.74 16.07* 39.80 17.95 0.51 32.50 6.45 0.33 49.88 28.44 0.04 17.65 Note: ND: lower than limit of quantitation: *a group of measured values, no SD value

(18) TABLE-US-00002 TABLE 2 Pharmacokinetic parameters (Mean SD, n = 3) calculated as tizoxanide, after oral administration of nitazoxanide and Compound 1, 2, 3, 4, 5, 6 in mice Parameter values Parameters Unit NTZ 1 2 3 4 5 6 T.sub.max h 0.25 0.58 0.14 0.83 1.00 0.08 1.70 0.00 0.38 0.10 0.29 0.00 0.00 0.55 C.sub.max ng/mL 329.4 681.2 506.5 875.2 705.8 329.8 430.11 111.2 194.5 144.6 90.8 113.3 37.9 155.20 Ke 1/h 0.41 0.36 0.58 0.35 0.48 0.37 0.32 0.23 0.08 0.16 0.10 0.21 0.11 0.05 t.sub.1/2 h 2.02 2.01 1.27 2.13 1.76 1.95 2.34 0.92 0.45 0.38 0.73 1.06 0.59 0.37 AUC.sub.0-t h*ng/mL 185.8 1176.8 902.3 2726.3 2197.7 193.9 2160.7 18.3 354.8 381.3 386.5 263.8 14.7 766.6 AUC.sub.0- h*ng/mL 198.5 1190.5 920.4 2883.7 2272.2 195.3 2345.5 28.5 353.0 388.8 611.2 359.6 15.1 643.4 MRT.sub.0-t h 1.46 2.22 1.78 2.85 2.47 1.52 3.22 0.81 0.30 0.51 0.37 0.76 0.17 0.55 MRT.sub.0- h 2.15 2.38 1.98 3.44 2.84 1.62 3.85 1.66 0.24 0.59 1.17 1.37 0.15 0.49 F.sub.(rt) % 633.29 485.59 1467.20 1182.73 104.35 1162.92 190.95 205.18 208.01 141.94 7.93 418.90

(19) The pharmacokinetic screening results after oral administration in mice show:

(20) (1) Compound 1, Compound 3, Compound 4 and Compound 6 were much better than the control agent nitazoxanide (NTZ) with respect to oral absorption; said four Compounds (calculated as tizoxanide) had a C.sub.max 2-fold, 2.7-fold, 2.14-fold and 1.31-fold higher than that of nitazoxanide, respectively; had a T.sub.max of 0.58 h, 0.83 h, 1.00 h and 1.7 h, respectively (which were obviously delayed compared to nitazoxanide with a time to peak (T.sub.max) of 0.25 h); had a Mean Retention Time (MRT) of 2.38 h, 3.44 h, 2.84 h and 3.22 h calculated as tizoxanide, respectively (which were obviously delayed compared to nitazoxanide with a MRT of 2.15 h); had an Area Under concentration-time Curve (AUC) of 1190.5 h*ng/mL, 2883.7 h*ng/mL, 2272.2 h*ng/mL, and 2160.7*ng/mL calculated astizoxanide, respectively (which were greatly increased compared to 198.5 h*ng/mL of nitazoxanide), and were 6.3-fold, 14.7-fold, 11.8-fold and 11.62-fold of that of nitazoxanide (i.e., their relative bioavailability calculated as tizoxanide was 6.3-fold, 14.7-fold, 11.8-fold and 11.62-fold of that of nitazoxanide, respectively).

(21) (2) Compound 2 was faster than the control agent nitazoxanide (NTZ) with respect to oral absorption, and had a T.sub.max of 0.14 h, which was smaller than 0.25 h of nitazoxanide; Compound 2 had a C.sub.max 1.5-fold of that of nitazoxanide calculated as tizoxanide; Compound 5 was comparable to nitazoxanide with respect to C.sub.max value calculated as tizoxanide; after oral administration of Compound 2, Compound 2 had an AUC of 920.4 h*ng/mL calculated as tizoxanide, which was greatly increased compared to 198.5 h*ng/mL of nitazoxanide, and was 4.9-fold of that of nitazoxanide (i.e., the relative bioavailability calculated as tizoxanide was 4.9 fold of that of nitazoxanide).

(22) (3) Compound 5 was faster than the control agent nitazoxanide (NTZ) with respect to oral absorption, but was comparable to nitazoxanide with respect to parameters such as T.sub.max, C.sub.max, MRT and AUC calculated astizoxanide.

(23) Conclusion: Compound 1, 2, 3, 4, 6, were significantly superior to the control agent nitazoxanide (NTZ) with respect to parameters such as C.sub.max, Mean Retention Time (MRT), Area Under concentration-time Curve (AUC) and relative bioavailability (F.sub.rt) calculated as tizoxanide. Compound 5 was faster than the control agent nitazoxanide (NTZ) with respect to oral absorption, but was comparable to nitazoxanide with respect to parameters such as T.sub.max, C.sub.max, MRT and AUC calculated as tizoxanide.

(24) Although the embodiments of the invention have been described in detail, a person skilled in the art would understand that a variety of modifications and replacements may be performed to the details according to all the teachings disclosed therein. These changes all fall into the protection scope of the invention. The scope of the invention is defined by the claims and any equivalent thereof.