Y-type discrete polyethylene glycol derivative and preparation method thereof

Abstract

The present invention discloses a Y-type discrete polyethylene glycol derivative, which has the advantages of determined molecular weights and the number of chain segments, and can avoid the defect of heterogeneity of a PEG derivative. In addition, the Y-type discrete polyethylene glycol derivative of the present invention may increase the water solubility of the discrete polyethylene glycol, and solve the problem of insufficient water solubility of the discrete polyethylene glycol-modified insoluble drug caused by an increase of the loading capacity.

Claims

1. A Y-type discrete polyethylene glycol derivative having the structure of formula (I): ##STR00023## wherein: in and n are integer of 0-30; A and B are the same or different YX structure; X is a linking group selected from the group consisting of: (CH.sub.2).sub.i, (CH.sub.2).sub.iNH, (CH.sub.2).sub.iOCOO, (CH.sub.2).sub.iOCONH, (CH.sub.2).sub.iNHCONH, OC(CH.sub.2).sub.iCOO, (CH.sub.2).sub.iCOO and (CH.sub.2).sub.iCONH, i is an integer from 0 to 10; Y is a reactive end group selected from the group consisting of C1-C6 alkoxy, hydroxy, amino, aminomethyl, maleimide, carboxy, mercapto group, succinimide carbonate, succinimide acetate, succinimide propionate, succinimide succinate, succinimide, dithiopyridyl, propionic acid, aldehyde group, thioester group, acryloxy, azido, glutaric acid, hydrazide, alkynyl, p-nitrophenyl carbonate, isocyanato, silane, and carboxymethyl; E.sub.1 is a discrete polyethylene glycol group with a structure of (CH.sub.2CH.sub.2O).sub.j, and j is an integer of 1 to 100; and, E.sub.2 is a discrete polyethylene glycol group with a structure of (CH.sub.2CH.sub.2O).sub.k, and k is an integer of 1 to 100.

2. The Y-type discrete polyethylene glycol derivative of claim 1, wherein in is an integer of 2 to 10, and/or, n is an integer of 2 to 10.

3. The Y-type discrete polyethylene glycol derivative of claim 2, wherein in is 2, 3, 4 or 5, and/or, n is 2, 3, 4 or 5.

4. The Y-type discrete polyethylene glycol derivative of claim 1, wherein the linking group X is (CH.sub.2).sub.i, (CH.sub.2).sub.iNH or (CH.sub.2).sub.iCONH.

5. The Y-type discrete polyethylene glycol derivative of claim 1, wherein for the linking group X, i is 0, 1, 2, 3 or 4.

6. The Y-type discrete polyethylene glycol derivative of claim 1, wherein the reactive group Y is selected from the group consisting of methoxy, hydroxy, amino, mercapto, carboxy, ester, aldehyde group, acrylic or maleimide.

7. The Y-type discrete polyethylene glycol derivative of claim 1, wherein for the discrete polyethylene glycol group E.sub.1, j is an integer of 1 to 20; and/or, for the discrete polyethylene glycol group E.sub.2, k is an integer of 1 to 20.

8. The Y-type discrete polyethylene glycol derivative of claim 7, wherein j is an integer of 1 to 12, and/or, k is an integer of 1 to 12.

9. The Y-type discrete polyethylene glycol derivative of claim 8, wherein j is 1, 2, 3, 4, 5, 6, 7 or 8; and/or, k is 1, 2, 3, 4, 5, 6, 7 or 8.

10. The Y-type discrete polyethylene glycol derivative of claim 1, wherein the Y-type discrete polyethylene glycol derivative has the following structure: ##STR00024##

11. The Y-type discrete polyethylene glycol derivative of claim 1, wherein the Y-type discrete polyethylene glycol derivative has the following structure with a structure of: ##STR00025##

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1 is a synthetic route diagram of Y-type discrete polyethylene glycol derivative (mEG.sub.3).sub.2NC.sub.3H.sub.6OH.

(2) FIG. 2 is a synthetic route diagram of Y-type discrete polyethylene glycol derivative (mEG.sub.5).sub.2NC.sub.3H.sub.6OH.

(3) FIG. 3 is a synthetic route diagram of Y-type discrete polyethylene glycol derivative (HO-EG.sub.4).sub.2NC.sub.3H.sub.6OH.

(4) FIG. 4 is a synthetic route diagram of Y-type discrete polyethylene glycol derivative (NH.sub.2-EG.sub.4).sub.2NC.sub.3H.sub.6OH.

(5) FIG. 5 is a synthetic route diagram of Y-type discrete polyethylene glycol derivative (HOOC-EG.sub.4).sub.2NC.sub.2H.sub.4NH.sub.2.

(6) FIG. 6 is a synthetic route diagram of Y-type discrete polyethylene glycol derivative (mEG.sub.3).sub.2N-EG.sub.4-OH.

(7) FIG. 7 shows the results of a dispersibility comparison test of cholesterol and cholesterol derivatives in aqueous solution (from the left to the right are the aqueous solutions of cholesterol derivative of (mEG.sub.3).sub.2NC.sub.3H.sub.6OH, cholesterol derivative of mEG.sub.7-OH and cholesterol with the same concentration, respectively.)

DETAILED DESCRIPTION OF THE INVENTION

Example 1: Synthesis of (mEG3)2NC3H6OH

(8) The synthetic route is shown in FIG. 1.

(9) 1. Synthesis of mEG.sub.3-OMs

(10) TEA (32 mL, 230 mmol) and 150 mL DCM were added to mEG.sub.3-OH (32 mL, 200 mmol), and the resulting mixture was placed in a reaction flask in an ice-water bath. MsCl (17.5 mL, 220 mmol) was dissolved with DCM (50 mL), and when dissolved completely, the resulting mixture was dropwise added into the reaction flask in an ice-water bath. The reaction was carried out at room temperature for 3 hours. Whether the reaction was complete was detected by thin layer chromatography (TLC). The reaction mixture was washed three times with water (150 mL). The organic phase was dried over anhydrous sodium sulfate and the sodium sulfate was removed by filtration. About 52 g product was obtained after concentration.

(11) 2. Synthesis of YType Small Molecule PEG

(12) mEG.sub.3-OMs (21.6 g, 89.3 mmol) prepared in the above step 1 and THF (150 mL) were added into aminopropanol (3.1 g, 41.3 mmol). After heating and refluxing overnight, the supernatant solution was poured and evaporated to dryness to obtain a crude product, which was purified by a column (250 g silica gel, the mobile phase was MeOH/DCM system, MeOH/DCM=3-7%) to give the product in a yield of 2.5 g (16%).

(13) NMR(CDCl.sub.3) : 3.5-3.8 (m, 22H, OCH.sub.2), 3.37 (s, 6H, CH.sub.3O), 2.7-2.8 (m, 6H, N(CH.sub.2).sub.3, 1.6-1.7 (m, 2H, NCH.sub.2CH.sub.2CH.sub.2OH); ESI-MS: 368.3 (M+H).sup.+, 390.2 (M+Na).sup.+.

Example 2: Synthesis of (mEG5)2NC3H6OH

(14) The synthetic route is shown in FIG. 2.

(15) 1. Synthesis of mEG.sub.5-OMs

(16) TEA (6.86 mL, 48 mmol) and DCM (60 mL) were added to mEG.sub.5-OH (10.08 g, 40 mmol), and the resulting mixture was placed in a reaction flask in an ice-water bath. MsCl (3.56 mL, 46 mmol) was dissolved with DCM (40 mL), and the resulting mixture was dropwise added into the reaction flask in an ice-water bath. The reaction was carried out at room temperature for 3 hours. Whether the reaction was complete was detected by TLC. The reaction mixture was washed three times with water (150 mL). The organic phase was dried over anhydrous sodium sulfate and the sodium sulfate was removed by filtration. About 10 g product was obtained after concentration.

(17) 2. Synthesis of YType Small Molecule PEG

(18) Aminopropanol (0.45 g, 6.06 mmol), TEA (1 mL) and THF (100 mL) were added into the mEG.sub.5-OMs (4 g, 12.1 mmol) prepared in the above step 1, after heating and refluxing overnight, the supernatant solution was poured and evaporated to dryness to obtain a crude product, which was purified by a column (the mobile phase was MeOH/DCM system, MeOH/DCM=3-7%) to give the product in a yield of 0.3 g (9.1%).

(19) NMR(CDCl.sub.3) : 3.5-3.8 (m, 38H, OCH.sub.2), 3.37 (s, 6H, CH.sub.3O), 2.7-2.8 (m, 6H, N(CH.sub.2).sub.3, 1.6-1.7 (m, 2H, NCH.sub.2CH.sub.2CH.sub.2OH); ESI-MS: 544.4 (M+H).sup.+, 566.3 (M+Na).sup.+.

Example 3: Synthesis of (HO-EG4)2-C3H6OH

(20) The synthetic route is shown in FIG. 3.

(21) Aminopropanol (3.06 g, 40.9 mmol) and THF (150 mL) were added to HO-EG.sub.4-Br (21 g, 81.7 mmol), after heating and refluxing overnight, the resulting mixture was cooled and evaporated to remove the solvent to obtain a crude product, which was purified by a column (the mobile phase was MeOH/DCM system, MeOH/DCM=0-10%) to give the product in a yield of 2.3 g (13.1%).

(22) NMR(CDCl.sub.3, hydrochloride) : 3.4-3.9 (m, 28H, OCH.sub.2), 3.3 (m, 2H, CH.sub.2CH.sub.2CH.sub.2OH), 3.1-3.2 (m, 6H, NCH.sub.2), 2.0 (m, 2H, CH.sub.2CH.sub.2CH.sub.2OH); ESI-MS: 428.4 (M+H).sup.+.

Example 4: Synthesis of (NH2-EG4)2NC3H6OH

(23) The synthetic route is shown in FIG. 4.

(24) 1. Synthesis of N.sub.3-EG.sub.4-OH

(25) EG.sub.4 (36 mL, 210 mmol), DCM (100 mL) and TEA (25 mL) were added to a reaction flask, which was placed in an ice-water bath. The resulting mixture was dropwise added with a DCM solution (100 mL) containing MsCl (5.81 mL, 75 mmol). The reaction was carried out at room temperature for 4 hours. The reaction mixture was washed once with water (100 mL) and evaporated to dryness to give a crude product.

(26) 95% ethanol (150 mL) and sodium azide (6.5 g, 100 mmol) were added to the crude product obtained in the previous step. The resulting mixture was refluxed at room temperature for 16 hours. The solid was removed by filtration, and the solution was evaporated to dryness. The substance obtained after evaporation was added with DCM (150 mL) and washed three times with water (100 mL). The DCM phase was evaporated to dryness to give 14 g crude product, which was purified by a column (250 g silica gel, the mobile phase was PE/EA system, PE/EA=50-0%) to give the product in a yield of 12.3 g (26.7%).

(27) 2. Synthesis of N.sub.3-EG.sub.4-OMs

(28) TEA (8.6 mL, 60.3 mmol, 1.2 eq) and DCM (150 mL) were added into N.sub.3-EG.sub.4-OH (11 g, 50.2 mmol) prepared in the above step 1. The resulting mixture was placed in an ice-water bath. MsCl (4.5 mL, 57.8 mmol, 1.15 eq) was dissolved with DCM (50 mL), and the resulting mixture was added dropwise to the reaction flask in ice-water bath. The reaction was carried out overnight at room temperature. Whether the reaction was complete was detected by TLC. The reaction mixture was washed three times with water (100 mL). The organic phase was dried over anhydrous sodium sulfate and the sodium sulfate was removed by filtration. The product was obtained after concentration.

(29) 3. Synthesis of (N.sub.3-EG.sub.4).sub.2NC.sub.3H.sub.6OH

(30) Aminopropanol (1.86 mL, 24.4 mmol) and DMF (130 mL) were added into N.sub.3-EG.sub.4-OMs (14.5 g, 48.8 mmol) prepared in the above step 2, after heating and refluxing overnight, the resulting mixture was evaporated to dryness to obtain a crude product, which was purified by a column (the mobile phase was MeOH/DCM system, MeOH/DCM=3-7%) to give the product in a yield of 1.2 g (10.3%).

(31) 4. Synthesis of (NH.sub.2-EG.sub.4).sub.2NC.sub.3H.sub.6OH

(32) DMF (20 mL) and triphenylphosphine (615 mg, 2.35 mmol, 1.4 eq*2) were added to (N.sub.3-EG.sub.4).sub.2NOH (400 mg, 0.84 mmol) prepared in the above step 3, and after reacting overnight at room temperature, the resulting mixture was added with water (0.1 mL), then the reaction was carried out overnight. The resulting mixture was evaporated to remove DMF, added with water (50 mL), successively washed twice with toluene (40 mL) and DCM (30 mL), and evaporated to remove moisture to give the product in a yield of 300 mg (84%).

(33) NMR (D.sub.2O) : 3.5-3.8 (m, 38H, OCH.sub.2), 2.4-2.8 (m, 10H, N(CH.sub.2).sub.3 NH.sub.2CH.sub.2, 1.6-1.7 (m, 2H, NCH.sub.2CH.sub.2CH.sub.2OH); ESI-MS: 448.4 (M+Na).sup.+.

Example 5: Synthesis of (HOOC-EG4)2NC2H4NH2

(34) The synthetic route is shown in FIG. 5.

(35) 1. Synthesis of MsO-EG.sub.4-OtBu

(36) DCM (350 mL) and TEA (22.7 mL, 0.159 mol, 1.3 eq) were added to HO-EG.sub.4-tBu (39.4 g, 0.122 mol). The resulting mixture was added with a solution of MsCl (11.4 mL, 0.146 mol, 1.2 eq) in DCM (150 mL) under ice-water bath. The reaction was carried out overnight at room temperature. The resulting mixture was successively washed with water (200 mL) three times, dried over anhydrous sodium sulfate, filtered and evaporated to remove the solvent to give the product.

(37) 2. Synthesis of (tBuO-EG.sub.4).sub.2NC.sub.2H.sub.4NH-Boc

(38) NH.sub.2C.sub.2H.sub.4NH-Boc (4 g, 25 mmol) and THF (200 mL) were added to tBu-EG.sub.4-OMs (20 g, 50 mmol) prepared in the above step 1, the resulting mixture was stirred overnight at room temperature and concentrated to obtain a crude product, which was purified by a column (MeOH/DCM=0-10%) to give the product in a yield of 2.3 g (12.0%).

(39) 3. Synthesis of (HOOC-EG.sub.4).sub.2NC.sub.2H.sub.4NH.sub.2

(40) DCM (20 mL) and trifluoroacetic acid (TFA) (8 mL) were added into (tBuO-EG.sub.4).sub.2NC.sub.2H.sub.6NH-Boc (2.3 g, 3.0 mmol) prepared in the above step 2. The reaction was carried out overnight at room temperature. The resulting mixture was evaporated to remove the solvent to obtain the product in a yield of 2.2 g (94.0%). NMR(CDCl.sub.3) : 2.6-2.7 (t, 4H, CH.sub.2COO); 3.5-3.8 (m, 40H, other hydrogen); ESI-MS: 557.4 (M+H).sup.+, 595.3 (M+Na).sup.+.

Example 6: Synthesis of (mEG3)2N-EG4-OH

(41) 1. Synthesis of NH.sub.2-EG.sub.4-OH

(42) THF (50 mL) was added to N.sub.3-EG.sub.4-OH (9.2 g, 42.0 mmol). The resulting mixture was placed in an ice-water bath, and added dropwise with THF (75 mL) containing triphenylphosphine (13.21 g, 50.4 mmol, 1.2 eq). The reaction was carried out at room temperature for 24 hours. The resulting mixture was added with water (1965 uL, 109.2 mmol, 2.6 eq), and reacted at room temperature overnight. The resulting mixture was evaporated to remove the solvent, added with water (150 mL), washed with toluene (150 mL) and DCM (100 mL), and evaporated to remove the water phase to give 8 g product.

(43) 2. Synthesis of (mEG.sub.3).sub.2N-EG.sub.4-OH

(44) NH.sub.2-EG.sub.4-OH (4.3 g, 22.3 mmol) prepared in step 1 and THF (150 mL) were added to EG.sub.3-OMs (10.8 g, 44.6 mmol). The resulting mixture was refluxed and stirred overnight, and evaporated to remove the solvent to obtain a crude product, which was purified by a column (the mobile phase was MeOH/DCM system, MeOH/DCM=3-7%) to give the product in a yield of 2.1 g (19.4%). NMR(CDCl.sub.3) : 3.5-3.8 (m, 34H, OCH.sub.2), 3.37 (s, 6H, CH.sub.3O), 2.7-2.8 (m, 6H, N(CH.sub.2).sub.3; ESI-MS: 486.4 (M+H).sup.+, 508.4 (M+Na).sup.+.

Example 7: Synthesis of Cholesterol Derivative of (mEG3)2NC3H6OH

(45) DCM (20 mL) and TEA (0.905 mL, 6.53 mmol) were added to the (mEG.sub.3).sub.2NC.sub.3H.sub.6OH (2 g, 5.45 mmol) prepared in Example 1. The cholesterol chloroformate (2.57 g, 5.72 mmol) was dissolved in DCM (30 mL), and the resulting mixture was added dropwise to the reaction flask. The resulting mixture was stirred overnight at room temperature. Whether the reaction was complete was detected by TLC. The resulting mixture was washed once with water, and spin-dried to obtain a crude product, which was purified by a column (the mobile phase was MeOH/DCM system, MeOH/DCM=0-7%) to give 1.4 g product (32.4%).

(46) NMR(CDCl.sub.3) : 5.55-5.45 (m, 1H), 4.65-4.50 (m, 1H), 4.1-4.0 (m, 2H), 3.5-3.8 (m, 20H), 3.37 (s, 6H), 2.7-2.8 (m, 6H), 2.50-0.80 (m, 42H), 0.65-0.60 (m, 3H).

Example 8: Synthesis of Cholesterol Derivative of mEG7-OH

(47) A cholesterol derivative of mEG.sub.7-OH was prepared under the same conditions as in Example 7 by commercially available mEG.sub.7-OH.

(48) NMR(CDCl.sub.3) : 5.55-5.45 (m, 1H), 4.65-4.50 (m, 1H), 4.1-4.0 (m, 2H), 3.5-3.8 (m, 26H), 3.37 (s, 3H), 2.50-0.80 (m, 40H), 0.65-0.60 (m, 3H).

Example 9: Comparison Test of Dispersion of Cholesterol and Cholesterol Derivatives in Water

(49) 20 mg cholesterol derivative of (mEG.sub.3).sub.2NC.sub.3H.sub.6OH prepared in Example 7, 20 mg cholesterol derivative of mEG.sub.7-OH prepared in Example 8 and 20 mg cholesterol were placed in a 50 mL volumetric flask, which was added with water to the scale, and placed in 20 C. water bath, shaken vigorously every 5 min for 30 min. After 30 min, the dispersion of the two cholesterol derivatives and cholesterol in water were shown in FIG. 7, from the left to the right are successively cholesterol derivative of (mEG.sub.3).sub.2NC.sub.3H.sub.6OH, cholesterol derivative of mEG.sub.7-OH and cholesterol. The results with the naked eye observation showed that the dispersion liquid of cholesterol derivative of (mEG.sub.3).sub.2NC.sub.3H.sub.6OH was translucent light blue, and the handwriting behind the volumetric flask can be clearly seen; and the cholesterol derivative of mEG.sub.7-OH was a white emulsion, and cholesterol is completely insoluble.

Example 10: Comparison Test of Solubility of Cholesterol Derivatives in Water

(50) The solubility of cholesterol derivatives in water was determined according to the method in Chinese pharmacopoeia:

(51) 21.3 mg cholesterol derivative of (mEG.sub.3).sub.2NC.sub.3H.sub.6OH prepared in Example 7 was dispersed in 50 mL water. 5 mL was taken out therefrom, and added with 1.5 mL water, the resulting mixture was placed in a 20 C. water bath, and shaken vigorously every 5 min for 30 min. After 30 min, 1.5 mL water was further added, and the resulting mixture was placed in a 20 C. water bath with the same procedure for dissolution. Until 9 mL water was added, the solution was clear.

(52) 24.2 mg cholesterol derivative of mEG.sub.7-OH prepared in Example 8 was dispersed in 50 mL water. 5 mL was taken out therefrom, and added with 5 mL water, the resulting mixture was placed in a 20 C. water bath, and shaken vigorously every 5 min for 30 min. After 30 min, 5 mL water was further added, and the resulting mixture was placed in a 20 C. water bath with the same procedure for dissolution until 25 mL water was added. 5 mL solution was taken from the dilute solution, the same procedure was carried out until 10 mL water was added, and the solution was clear.

(53) From the results analysis, it was found that the solubility of cholesterol derivative of (mEG.sub.3).sub.2NC.sub.3H.sub.6OH was 15.2 mg/100 g, which was 3.3 times the solubility of cholesterol derivative of mEG.sub.7-OH, 4.61 mg/100 g, and was 76 times the solubility of cholesterol (<0.2 mg/100 g).