LIVER TARGETING DRUG, PHARMACEUTICAL COMPOSITION AND USE THEREOF

Abstract

Disclosed are a compound represented by formula I or a pharmaceutically acceptable salt thereof, and a pharmaceutical composition and use thereof. The compound has significant liver targeting characteristics, and can reduce the drug concentration in the circulating system while improving the efficacy, thereby reducing the toxic and side effects.

##STR00001##

Claims

1. A compound represented by formula I: ##STR00020## or a pharmaceutically acceptable salt thereof, wherein: R.sup.1 and R.sup.2 are each independently selected from: hydrogen, —COR′, —CONR′R″, or —COOR′, where R.sup.1 and R.sup.2 are not hydrogen at the same time; optionally, R.sup.1 and R.sup.2 are each independently selected from: hydrogen or —COR′, where R.sup.1 and R.sup.2 are not hydrogen at the same time; R′, at each occurrence, and R″ are each independently selected from: hydrogen, alkyl or substituted alkyl, alkenyl or substituted alkenyl, alkynyl or substituted alkynyl, cycloalkyl or substituted cycloalkyl, aryl or substituted aryl, heteroaryl or substituted heteroaryl, heterocyclyl or substituted heterocyclyl; and substituents of each of the substituted alkyl, substituted alkenyl, substituted alkynyl, substituted cycloalkyl, substituted aryl, substituted heteroaryl, or substituted heterocyclyl are each independently selected from: halogen, cyano, amino, nitro, hydroxyl, alkyl, alkoxyl, modified alkyl, and amino acidyl.

2. The compound of formula I, or the pharmaceutically acceptable salt thereof according to claim 1, wherein optionally, the “halogen” is selected from: F, Cl, Br, or I; optionally, “alkyl” in the “alkyl” or “alkoxyl” is C.sub.1-C.sub.20 linear or branched alkyl, optionally C.sub.1-C.sub.10 linear or branched alkyl, optionally C.sub.1-C.sub.5 linear or branched alkyl, or optionally selected from: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, and n-pentyl; optionally, the “modified alkyl” is a group resulting from substitution of one or more groups selected from —O—, —CO—, —NH.sub.2, —OH, halogen, —CN, and —NO.sub.2 for any carbon atom in the alkyl; optionally, the “cycloalkyl” is C.sub.3-C.sub.10 monocyclic or bicyclic cycloalkyl; optionally, C.sub.3-C.sub.7 monocyclic or bicyclic cycloalkyl; optionally, C.sub.5-C.sub.6 monocyclic or bicyclic cycloalkyl; optionally, the aryl is 6- to 14-membered aryl; optionally, 6- to 10-membered aryl; optionally, phenyl or naphthyl; optionally, the “heteroaryl” is a 5- to 14-membered heteroaromatic ring containing 1, 2, or 3 heteroatoms selected from N, O, and S; optionally a 5- to 10-membered heteroaromatic ring; and optionally, the “heterocyclyl” is a 3- to 10-membered nonaromatic heterocyclic ring containing 1, 2, or 3 heteroatoms selected from N, O, and S.

3. The compound of formula I, or the pharmaceutically acceptable salt thereof according to claim 1, wherein optionally, R′, at each occurrence, and R″ are each independently selected from: hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, benzyl, sec-butyl, n-pentyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, phenyl and substituted phenyl, wherein a substituent of the phenyl is selected from F, Br, Cl, I, cyano, amino, nitro, hydroxyl, C.sub.1-C.sub.5 linear or branched alkyl, C.sub.1-C.sub.5 linear or branched alkoxyl, and natural amino acidyl; and optionally, R.sup.1 and R.sup.2 are each independently selected from: formyl, acetyl, propionyl, isopropionyl, butyryl, isobutyryl, benzoyl, and substituted benzoyl, wherein a substituent of the phenyl is selected from F, Br, Cl, I, cyano, amino, nitro, hydroxyl, C.sub.1-C.sub.5 linear or branched alkyl, C.sub.1-C.sub.5 linear or branched alkoxyl, and natural amino acidyl.

4. The compound of formula I, or the pharmaceutically acceptable salt thereof according to claim 1, wherein the pharmaceutically acceptable salt includes a salt formed of a compound of formula I and an acid; optionally, the acid includes an inorganic acid and an organic acid; optionally, the inorganic acid includes hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, phosphoric acid, and carbonic acid; and optionally, the organic acid includes formic acid, ascorbic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, citric acid, citric acid, tartaric acid, gluconic acid, hydrogen tartaric acid, glucuronic acid, carbonic acid, picric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzoic acid, benzenesulfonic acid, p-bromobenzenesulfonic acid, glutamic acid, salicylic acid, and pamoic acid; and the pharmaceutically acceptable salt is hydrochloride.

5. The compound of formula I, or the pharmaceutically acceptable salt thereof according to claim 1, which selected from the following compounds: ##STR00021## ##STR00022## ##STR00023## ##STR00024##

6. The compound of formula I, or the pharmaceutically acceptable salt thereof according to claim 5, which is a compound I-5-S in an amorphous form, and optionally, a compound I-5-S having a CuKα-XPRD diffraction pattern as shown in FIG. 1.

7. A pharmaceutical composition, comprising one or more of a compound of formula I, or a pharmaceutically acceptable salt thereof according to claim 1, and optionally a pharmaceutically acceptable carrier.

8. The pharmaceutical composition according to claim 7, wherein a dosage form of the pharmaceutical composition includes an oral preparation and an injection preparation; optionally, the oral preparation includes a solid preparation and a liquid preparation; optionally, the solid preparation includes tablets, powders, granules, and capsules; and optionally, the liquid preparation includes water or oil suspensions, and syrups.

9. Use of a compound of formula I or a pharmaceutically acceptable salt thereof according to claim 1, or a pharmaceutical composition in the preparation of a medicament for treating liver diseases; optionally, the liver diseases include liver cancer and hepatitis B.

10. Use of a compound of formula I or a pharmaceutically acceptable salt thereof according to claim 1, or a pharmaceutical composition in the preparation of a medicament for treating hysteroma, lymphoid leukemia, solid tumors, myelodysplasia syndrome, or head and neck tumors.

11. Use of a compound of formula I or a pharmaceutically acceptable salt thereof according to claim 1, or a pharmaceutical composition in the preparation of a medicament for inhibiting an action of an apoptosis protein.

12. Use of a compound of formula I or a pharmaceutically acceptable salt thereof according to claim 1, or a pharmaceutical composition in the preparation of a medicament for use in an immune system to fight against cancers.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0040] FIG. 1 is a CuKα-XPRD diffraction pattern of the compound I-5-S in Example 1.

DETAILED DESCRIPTION

[0041] The specific embodiments of the present disclosure will be elaborated below. It should be appreciated that the specific embodiments described herein are merely intended to exemplarily illustrate, and not limit the present disclosure.

Example 1. Preparation of Compounds

[0042] (1) Preparation of Compound I-5-1:

##STR00008##

[0043] Birinapant (400 mg, 0.50 mmol), di-tert-butyl dicarbonate (273 mg, 1.25 mmol), and triethylamine (152 mg, 1.50 mmol) were dissolved in THF (10 mL), reacted overnight at room temperature, and extracted with water (10 mL) and ethyl acetate (20 mL). The organic phase was washed with a saturated salt solution, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica-gel column chromatography (DCM:MeOH=20:1) to give a white solid I-5-1 (412 mg, 83%).

[0044] .sup.1H NMR (400 MHz, DMSO-.sub.d6) δ 11.92 (s, 2H), 8.17 (s, 2H), 7.83 (dd, J=8.7, 5.5 Hz, 2H), 7.46 (dd, J=9.8, 1.9 Hz, 2H), 6.93 (td, J=9.7, 2.3 Hz, 2H), 5.65 (d, J=3.0 Hz, 2H), 4.94-4.35 (m, 6H), 4.23 (s, 2H), 3.87 (dd, J=11.1, 4.4 Hz, 2H), 3.68 (d, J=10.8 Hz, 2H), 3.56-3.39 (m, 4H), 2.79 (s, 6H), 1.91-1.67 (m, 8H), 1.40 (s, 18H), 1.35 (dd, J=10.1, 4.7 Hz, 6H), 1.01 (s, 6H).

[0045] (2) Preparation of Compound I-5-2

##STR00009##

[0046] Compound I-5-1 (312 mg, 0.31 mmol), acetic anhydride (76 mg, 0.74 mmol), triethylamine (125 mg, 1.24 mmol), and DMAP (8 mg, 0.06 mmol) were dissolved in THF (10 mL), reacted overnight at room temperature, and extracted with water (10 mL) and ethyl acetate (20 mL). The organic phase was washed with a saturated salt solution, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica-gel column chromatography (PE:EA=1:1) to give a white solid I-5-2 (280 mg, 83%).

[0047] .sup.1H NMR (400 MHz, CDCl.sub.3) δ 11.56 (s, 2H), 7.54 (dt, J=10.1, 5.1 Hz, 4H), 6.88 (td, J=9.4, 2.3 Hz, 2H), 5.49 (t, J=5.1 Hz, 2H), 4.86-4.60 (m, 4H), 4.50 (t, J=9.3 Hz, 2H), 4.19 (d, J=7.6 Hz, 2H), 3.84 (d, J=12.4 Hz, 2H), 3.56 (dd, J=14.4, 2.2 Hz, 2H), 3.34 (dd, J=14.3, 12.0 Hz, 2H), 2.88 (s, 6H), 2.33 (s, 6H), 2.13-1.88 (m, 6H), 1.78 (tt, J=14.6, 7.3 Hz, 2H), 1.51 (s, 18H), 1.38 (d, J=7.2 Hz, 6H), 1.03 (t, J=7.3 Hz, 6H).

[0048] (3) Preparation of Compound I-5-S

##STR00010##

[0049] Compound I-5-2 (280 mg, 0.26 mmol) was dissolved in DCM (10 mL). HCl (g) was charged into the reaction mixture at room temperature. After the reaction was detected by LC-MS to be completed, the reaction mixture was filtered. The filter cake was washed with diethyl ether (5 mL×2), and drained to give a yellowish green solid I-5-S (198 mg, 80%).

[0050] .sup.1H NMR (400 MHz, DMSO-.sub.d6) δ 11.48 (s, 2H), 9.40-9.29 (m, 2H), 8.92 (dd, J=15.2, 7.2 Hz, 4H), 7.95 (dd, J=8.6, 5.6 Hz, 2H), 7.19 (dd, J=9.9, 2.2 Hz, 2H), 6.96 (td, J=9.8, 2.3 Hz, 2H), 5.12 (s, 2H), 4.53-4.30 (m, 4H), 3.96-3.80 (m, 4H), 3.66 (d, J=12.0 Hz, 2H), 3.49-3.29 (m, 2H), 2.89 (t, J=12.4 Hz, 2H), 2.49 (s, 6H), 2.48 (s, 6H), 1.90 (t, J=6.5 Hz, 2H), 1.79 (td, J=13.6, 6.8 Hz, 2H), 1.67 (dt, J=21.6, 7.5 Hz, 2H), 1.45-1.39 (m, 8H), 0.96 (t, J=7.4 Hz, 6H); m/z (ESI) [M−2HCl+Na].sup.+=913.5, [M−2HCl+H]=.sup.+891.5.

[0051] The CuKa-XPRD pattern of Compound I-5-S was tested as shown in FIG. 1, indicating that the Compound I-5-S was an amorphous crystal.

[0052] (4) Preparation of Compound I-5-R:

##STR00011##

[0053] Compound I-5-R was prepared by the same method for preparing the Compound I-5-S. The Compound I-5-2 was deprotected with methanesulfonic acid to obtain its mesylate I-5-R.

[0054] Deprotection with an acid could produce a corresponding salt. According to Example 1(4), the corresponding acid, acyl chloride or anhydride was used, and then deprotection was carried out with a corresponding acid to produce a corresponding salt of the target product I.

[0055] (5) Preparation of Compound I-5:

##STR00012##

[0056] Compound I-5-S (100 mg) was mixed with an aqueous sodium bicarbonate solution (2N, 20 mL). The mixture was extracted with dichloromethane (20 mL). The organic solution was dried over Na.sub.2SO.sub.4, and the solvent was evaporated to give a target product I-5.

[0057] (6) Preparation of Compound A:

##STR00013##

[0058] Birinapant (400 mg, 0.50 mmol) and pyridine (198 mg, 2.50 mmol) were dissolved in dichloromethane (5 mL). Chlorine benzyl formate (186 mg, 1.10 mmol) was dissolved in dichloromethane (5 mL), and added dropwise to the reaction system at 0° C. under nitrogen protection. After dropwise addition, the temperature was raised to room temperature, and the mixture was reacted overnight and extracted with water. The organic phase was washed with a saturated salt solution, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica-gel column chromatography (DCM:MeOH=10:1) to give 338 mg of yellowish solid A, with a yield of 75%.

[0059] .sup.1H NMR (400 MHz, DMSO) δ 11.96 (s, 2H), 8.35 (s, 2H), 7.83 (dd, J=8.6, 5.5 Hz, 2H), 7.48 (dd, J=9.8, 2.0 Hz, 2H), 7.44-7.26 (m, 10H), 6.93 (td, J=9.7, 2.2 Hz, 2H), 5.65 (d, J=2.8 Hz, 2H), 5.11 (s, 4H), 4.78 (s, 2H), 4.45 (d, J=24.2 Hz, 4H), 4.23 (s, 2H), 3.88 (d, J=7.0 Hz, 2H), 3.68 (d, J=11.0 Hz, 2H), 3.48 (dd, J=29.4, 14.0 Hz, 4H), 2.88 (s, 6H), 2.06-1.64 (m, 8H), 1.41 (s, 6H), 0.98 (s, 6H).

[0060] (7) Preparation of Compound B:

##STR00014##

[0061] Compound B was prepared by the method for preparing Compound A.

[0062] (8) Preparation of Compound I-6-S:

##STR00015##

[0063] Compound I-5-1 (400 mg, 0.50 mmol), dimethylcarbamyl chloride (1.25 mmol), and triethylamine (152 mg, 1.50 mmol) were dissolved in THF (10 mL), reacted overnight at room temperature, and extracted with water (10 mL) and ethyl acetate (20 mL). The organic phase was washed with a saturated salt solution, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica-gel column chromatography (DCM:MeOH=20:1) to give Compound C.

##STR00016##

[0064] Compound C (0.26 mmol) was dissolved in DCM (10 mL). HCl (g) was charged into the reaction mixture at room temperature. After the reaction was detected by LC-MS to be completed, the reaction mixture was filtered. The filter cake was washed with diethyl ether (5 mL×2), and drained to give a Compound I-6-S.

[0065] (9) Preparation of Compound I-7-S:

##STR00017##

[0066] Compound I-5-1 (400 mg, 0.50 mmol), n-octyl chloroformate (1.25 mmol), and triethylamine (152 mg, 1.50 mmol) were dissolved in THF (10 mL), reacted overnight at room temperature, and extracted with water (10 mL) and ethyl acetate (20 mL). The organic phase was washed with a saturated salt solution, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica-gel column chromatography (DCM:MeOH=20:1) to give Compound D.

##STR00018##

[0067] Compound D (0.26 mmol) was dissolved in DCM (10 mL). HCl (g) was charged into the reaction mixture at room temperature. After the reaction was detected by LC-MS to be completed, the reaction mixture was filtered. The filter cake was washed with diethyl ether (5 mL×2), and drained to give a Compound I-7-S.

Example 2. Pharmacokinetic Study

[0068] 1. Formulation of preparation and dosing: An appropriate amount of the test sample was precisely weighed, and mixed with an appropriate volume of a solvent (5% DMSO/5% Solutol/90% water) to obtain a clear solution or a homogeneous suspension. The preparation was administered to animals within 4 hours after formulation. The dosage formulation would be administered by oral gavage in accordance with facility standard operating procedures. The dosage volume would be determined by the body weights of the animals collected on the morning of administering.

[0069] 2. Blood collection: Blood (approx. 0.2 ml per point in time) would be drawn each time from the jugular vein of every animal. All blood samples would be transferred to pre-cooled commercial EDTA-K2 test tubes, and kept on wet ice until they were centrifuged.

[0070] 3. Plasma processing: Blood samples would be centrifuged (3200 rpm, 10 min) at approximately 4° C. Plasma was collected separately and transferred to pre-labeled PP tubes in wet ice at each point in time, and then immediately precipitated by ACN (6 IS) (the ratio of plasma: ACN was 1:4). Centrifugation (10 min, 12,000 rpm) was carried out again to obtain a supernatant. The supernatant was quickly frozen on dry ice and kept at −70±10° C. before LC/MS/MS analysis.

[0071] 4. Liver processing: Liver tissues were collected at each point in time, washed twice with pre-cooled deionized water, and dried with filter paper. Liver tissues were immediately homogenized with 10-times volume of a methanol-water solution (1:2, v/v). After homogenization, a portion of liver tissue homogenate (e.g., 200 uL of homogenate) was immediately measured. After precipitated on wet ice, the samples were centrifuged, and a supernatant was collected and stored in a refrigerator at −70±10° C. before LC-MS/MS analysis. From the remaining liver homogenate, 800 uL of the homogenate was measured as a spare.

[0072] Compounds Birinapant (5 mg/kg) and I-5-S (5.97 mg/kg) were intravenously injected (i.v.) into rats, respectively. Rat plasma and liver samples were collected at points in time of 0.25, 0.5, 1, 2, 4, 8, 12, 24 h, 48 h, and 72 h, respectively, and tested by LC-MS/MS according to the above-mentioned method to determine the concentrations of the in vivo active metabolites of Birinapant and I-5-S in plasma and liver (see Tables 1 and 2).

[0073] Compound I-5-S was metabolized into the active pharmaceutical ingredient Birinapant in animals:

##STR00019##

[0074] After the solution of compound Birinapant was intravenously injected into rats, the concentrations of the active compound Birinapant in the rats were determined in the plasma and liver, respectively (see Table 1).

[0075] After the solution of compound I-5-S was intravenously injected into rats, the concentrations of the active compound Birinapant in the rats were determined in the plasma and liver, respectively (see Table 2).

TABLE-US-00001 TABLE 1 Distribution of Drug Birinapant in Plasma and Liver after i.v. Birinapant in Rats PK Parameters Plasma Liver Rsq_adj 0.854 0.975 No points used for T.sub.1/2 5.00 6.00 C.sub.0(ng/mL) 3473 — C.sub.max (ng/mL or ng/g) 3473 4279 T.sub.max (h) — 0.250 T.sub.1/2 (h) 14.6 22.9 Vd.sub.ss (L/kg) 36.3 — Cl (mL/min/kg) 76.3 — T.sub.last (h) 48.0 72.0 AUC.sub.0-24 (ng .Math. h/mL) or (ng .Math. h/g) 967 40566 AUC.sub.0-last (ng .Math. h/mL) or (ng .Math. h/g) 1042 66712 AUC.sub.0-inf (ng .Math. h/mL) or (ng .Math. h/g) 1092 76641 MRT.sub.0-last (h) 4.99 23.2 MRT.sub.0-inf (h) 7.94 33.8 AUC.sub.Extra (%) 4.60 13.0 AUMC.sub.Extra (%) 40.1 40.2 AUC Ratio 64.0

TABLE-US-00002 TABLE 2 Distribution of Drug Birinapant in Plasma and Liver after i.v. I-5-S in Rats Birinapant PK Parameters Plasma Liver Rsq_adj 0.996 0.771 No points used for T.sub.1/2 4.00 4.00 C.sub.0(ng/mL) — — C.sub.max (ng/mL or ng/g) 205 3111 T.sub.max (h) 0.083 8.00 T.sub.1/2 (h) 35.3 32.6 Vd.sub.ss (L/kg) — — Cl (mL/min/kg) — — T.sub.last (h) 72.0 72.0 AUC.sub.0-last (ng .Math. h/mL) or (ng .Math. h/g) 504 97796 AUC.sub.0-inf (ng .Math. h/mL) or (ng .Math. h/g) 666 134133 MRT.sub.0-last (h) 27.6 27.7 MRT.sub.0-inf (h) 50.8 50.4 AUC.sub.Extra (%) 24.3 27.1 AUMC.sub.Extra (%) 58.9 61.5 AUC Ratio (L/P) 194

[0076] From the data listed in Tables 1 and 2, it could be seen that after a compound Birinapant solution was intravenously injected into rats, the initial concentration of the drug Birinapant in the plasma reached 3473 (ng.h/g), which would easily cause side effects on the circulating system. After a solution of the Birinapant prodrug, i.e., compound I-5-S was intravenously injected into rats, (1) the maximum concentration of the drug Birinapant in the plasma was reduced to 205 (ng/mL or ng/g), reducing by 94% in comparison to direct use of Birinapant; (2) a total drug exposure in the plasma was reduced by 49% in comparison to direct use of Birinapant; and (3) the total exposure of the active drug Birinapant released in liver reached 97796 (ng.h/g), which was 194 times that in the plasma and 2.4 times that of direct use of Birinapant.

[0077] The experimental results show that the compound represented by formula I of the present disclosure can effectively concentrate the drug in the liver, exhibiting a significant liver targeting characteristic, and can reduce the drug dosage while improving the therapeutic effects on liver diseases, and reduce the drug concentration in the circulating system, thereby reducing the toxic and side effects on the organs related to the circulating system. Reduction in side effects of the drug on the circulating system can effectively improve the drug tolerance in clinical, improve the therapeutic effects on other cancers, such as hysteroma, lymphoid leukemia, solid tumors, myelodysplasia syndrome, or head and neck tumors, and expand its clinical applications.

[0078] Under the same conditions as described above, after rats were orally or intravenously injected with a solution of Compound A or B, the concentrations of Compound A or B and drug Birinapant in rats were measured in plasma and liver, respectively. However, Compounds A and B, no matter whether administered orally or intravenously to rats, could not be degraded by in vivo esterase at all to release the active drug Birinapant, and Compounds A and B were 100% excreted and recovered. Therefore, such prodrugs would have no value of application for Birinapant.