Amphiphilic block copolymer, preparation method thereof and nanomicelle drug-loading system

Abstract

The present invention relates to an amphiphilic block copolymer represented by formula I, a preparation method thereof, and a nanomicelle drug delivery system formed from the copolymer and a poorly soluble drug. The amphiphilic block copolymer includes a hydrophilic chain segment, a hydrophobic chain segment, and a linker for linking the hydrophilic chain segment to the hydrophobic chain segment. The linker contains an unsaturated structure, which can enhance the interaction between the poorly soluble drug and the copolymer to improve the drug loading ability and stability of the nanomicelle. The invention also relates to a nanomicelle drug-loading system, a preparation method thereof, and the use of the nanomicelle drug-loading system for preparing medicines for treating tumors, inflammation, diabetes, central nervous system diseases, cardiovascular diseases, and psychological disorders. ##STR00001##

Claims

1. An amphiphilic block copolymer comprising a hydrophilic chain segment, a hydrophobic chain segment and a linker used to connect the hydrophilic chain segment and the hydrophobic chain segment, comprising the following structure: ##STR00102## wherein T.sup.3 is a single bond or ##STR00103## wherein Ar is an aromatic ring, wherein the aromatic ring is C.sub.6-C.sub.20 aryl, C.sub.6-C.sub.20 aryl substituted with R.sup.a, C.sub.2-C.sub.20 heteroaryl, or C.sub.2-C.sub.20 heteroaryl substituted with R.sup.b; wherein each of R.sup.a and R.sup.b is independently selected from C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, C.sub.4-C.sub.6 cycloalkyl, a halide, a hydroxyl or a nitro group; wherein a number of R.sup.a is one or more; wherein when the number of R.sup.a is more than one, R.sup.a is the same or different; wherein a number of R.sup.b is one or more; wherein when the number of R.sup.b is more than one, R.sup.b is the same or different; wherein a heteroatom in the C.sub.2-C.sub.20 heteroaryl or the R.sup.bsubstituted C.sub.2-C.sub.20 heteroaryl is O, S or N; wherein a number of heteroatom(s) is one or more; and wherein when the number of heteroatoms is more than one, the heteroatoms are the same or different; wherein the hydrophilic chain segment is a polyethylene glycol chain segment or a mono-protected polyethylene glycol chain segment with a number-average molecular weight ranging from 400 to 20000; and wherein the hydrophobic chain segment is one chain segment selected from the group consisting of a polylactide chain segment, a mono-protected polylactide chain segment, a polyglycolide chain segment, a mono-protected polyglycolide chain segment, a poly (lactide-co-glycolide) chain segment, a mono-protected poly (lactide-co-glycolide) chain segment, a polycaprolactone chain segment, a mono-protected polycaprolactone chain segment, a polycarbonate chain segment, a mono-protected polycarbonate chain segment, a polydioxanone chain segment and a mono-protected polydioxanone chain segment, with a number-average molecular weight ranging from 400 to 20000.

2. The amphiphilic block copolymer according to claim 1, wherein, in the aromatic ring, the C.sub.6-C.sub.20 aryl or the R.sup.a substituted C.sub.6-C.sub.20 aryl is a C.sub.6-C.sub.10 aryl; and/or, in the aromatic ring, the C.sub.2-C.sub.20 heteroaryl or the R.sup.b substituted C.sub.2-C.sub.20 heteroaryl is C.sub.2-C.sub.10 heteroaryl; and/or, in R.sup.a or R.sup.b, the C.sub.1-C.sub.6 alkyl is C.sub.1-C.sub.3 alkyl; and/or, in R.sup.a or R.sup.b, the C.sub.1-C.sub.6 alkoxy is C.sub.1-C.sub.3 alkoxy; and/or, the C.sub.1-C.sub.30 small molecular fragment is a C.sub.2-C.sub.10 small molecular fragment; and/or, the C.sub.1-C.sub.30 small molecular fragment is substituted by 1-3 aromatic rings; and/or, the C.sub.1-C.sub.30 small molecular fragment optionally comprises a heteroatom substitution; wherein the heteroatom is one or more heteroatom(s) selected from the group consisting of oxygen, nitrogen, sulfur and phosphorus; and wherein the number of the heteroatom(s) is one or more; and/or, the hydrophobic chain segment is selected from a polylactide chain segment or a mono-protected polylactide chain segment with a number-average molecular weight ranging from 400 to 2000.

3. The amphiphilic block copolymer according to claim 1, wherein the aromatic ring is the C.sub.6-C.sub.10 aryl, the R.sup.a substituted C.sub.6-C.sub.10 aryl, the C.sub.2-C.sub.10 heteroaryl or a R.sup.b substituted C.sub.2-C.sub.10 heteroaryl; and wherein each of R.sup.a and R.sup.b is independently selected from C.sub.1-C.sub.3 alkyl, C.sub.1-C.sub.3 alkoxy, the halide, the hydroxyl or the nitro group.

4. An amphiphilic block copolymer comprising a hydrophilic chain segment, a hydrophobic chain segment and a linker used to connect the hydrophilic chain segment and the hydrophobic chain segment, wherein the structure of the linker is a C.sub.1-C.sub.30 small molecular fragment derived from amino acid(s) containing an aromatic ring other than phenylalanine, from an amino alcohol containing the aromatic ring, or from a peptide containing the aromatic ring; wherein the aromatic ring is located in a side chain of the amino acid(s), amino alcohol or peptide, or is located in the protecting group of the hydroxy, thio, amino or carboxy functional group in the amino acid(s), amino alcohol or peptide, wherein the aromatic ring is C.sub.6-C.sub.20 aryl, C.sub.6-C.sub.20 aryl substituted with R.sup.a, C.sub.2-C.sub.20 heteroaryl, or C.sub.2-C.sub.20 heteroaryl substituted with R.sup.b; wherein each of R.sup.a and R.sup.b is independently selected from C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, C.sub.4-C.sub.6 cycloalkyl, halide, hydroxyl or nitro group; wherein a number of R.sup.a is one or more, and wherein when the number of R.sup.a is more than one, R.sup.a is the same or different; wherein a number of R.sup.b is one or more, and wherein when the number of R.sup.b is more than one, R.sup.b is the same or different; wherein a heteroatom in the C.sub.2-C.sub.20 heteroaryl or R.sup.b substituted C.sub.2-C.sub.20 heteroaryl is O, S or N; wherein a number of the heteroatom is one or more; and wherein when the number of the heteroatom is more than one, the heteroatoms is the same or different; wherein the hydrophilic chain segment is a polyethylene glycol chain segment or a mono-protected polyethylene glycol chain segment with a number-average molecular weight ranging from 400 to 20000; and wherein the hydrophobic chain segment is a one chain segment selected from the group consisting of a polylactide chain segment, a mono-protected polylactide chain segment, a polyglycolide chain segment, a mono-protected polyglycolide chain segment, a poly (lactide-co-glycolide) chain segment, a mono-protected poly (lactide-co-glycolide) chain segment, a polycaprolactone chain segment, a mono-protected polycaprolactone chain segment, a polycarbonate chain segment, a mono-protected polycarbonate chain segment, a polydioxanone chain segment and a mono-protected polydioxanone chain segment, with a number-average molecular weight ranging from 400 to 20000.

5. The amphiphilic block copolymer according to claim 4, wherein the amino acid(s) containing the aromatic ring is one or more amino acids selected from the group consisting of histidine, tyrosine, tryptophan and 3-(2-naphthyl)-alanine; and/or, wherein the amino alcohol containing the aromatic ring is one or more amino alcohol(s) selected from the group consisting of phenylalaninol, histidinol, tyrosinol, tryptosol and 3-(2-naphthyl)-alaninol; and/or, wherein one or more building blocks in the peptide containing the aromatic ring is derived from one or more amino acid(s) selected from the group consisting of phenylalanine, histidine, tyrosine, tryptophan and 3-(2-naphthyl)-alanine.

6. The amphiphilic block copolymer according to claim 1, wherein the amphiphilic block copolymer is: ##STR00104## wherein each of R.sup.1 and R.sup.2 is independently selected from a hydroxyl protecting group or hydrogen; wherein n=8-455; m=3-160; and wherein Ar and T.sup.3 are as defined in claim 1.

7. An amphiphilic block copolymer selected from any one of the group consisting of: ##STR00105## ##STR00106## ##STR00107## wherein n=8-455; and m=3-160.

8. An amphiphilic block copolymer selected from any one of the group consisting of: ##STR00108## wherein n=8-455; and m=3-160.

9. A method of preparing the amphiphilic block copolymer according to claim 1, comprising the following steps: 1) modifying polyethylene glycol or mono-protected polyethylene glycol with the linker; 2) in organic solvent, in the presence of catalyst, using the product from step 1) initiating polymerization of DL-lactide, L-lactide, or D-lactide, glycolide, a mixture of DL-lactide and glycolide with different ratios, a mixture of L-lactide and glycolide with different ratios, a mixture of D-lactide and glycolide with different ratios, caprolactone, a mixture of bisphenol A and diphenyl carbonate, or p-dioxanone; 3) optionally, protecting the terminal hydroxyl group of the polymer from step 2).

10. The method of claim 9, comprising the following steps: 1) in the presence of catalyst, initiating polymerization of lactide with a polymer of formula II to prepare a copolymer of formula IA, wherein the lactide is DL-lactide, L-lactide or D-lactide; ##STR00109## 2) conducting a hydroxyl protecting reaction of the copolymer of formula IA to prepare the amphiphilic block copolymer of formula I; ##STR00110## wherein the structure of the linker is the C.sub.1-C.sub.30 small molecular fragment substituted by one or more aromatic rings; wherein the one or more aromatic rings is independently selected from C.sub.6-C.sub.20 aryl, C.sub.6-C.sub.20 aryl substituted with R.sup.a, C.sub.2-C.sub.20 heteroaryl, or C.sub.2-C.sub.20 heteroaryl substituted with R.sup.b; wherein R.sup.a and R.sup.b are each independently selected from C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, C.sub.4-C.sub.6 cycloalkyl, halide, hydroxyl or nitro group; the number of R.sup.a is one or more; wherein when the number of R.sup.a is more than one, R.sup.a is the same or different; the number of R.sup.b is one or more; wherein when the number of R.sup.b is more than one, R.sup.b is the same or different; wherein the heteroatom in the C.sub.2-C.sub.20 heteroaryl or R.sup.b substituted C.sub.2-C.sub.20 heteroaryl is O, S or N; wherein the number of heteroatom(s) is one or more; wherein when the number of heteroatom(s) is more than one, the heteroatoms are the same or different; wherein R.sup.1 and R.sup.2 are each independently selected from hydroxyl protecting group or hydrogen; wherein n=8-455; m=3-160; and wherein when R.sup.2 is hydrogen, step 2) is not performed.

11. The method according to claim 10, wherein the method further comprises modifying the polymer of formula III with the small molecular fragment to prepare the polymer of formula II; ##STR00111## wherein the linker, R.sup.1 and n are as defined in claim 10.

12. The method according to claim 9, wherein the catalyst in step 2) is one or more catalysts selected from the group consisting of 1,8-diazabicyclo[5.4.0]undec-7-ene, stannous octoate, magnesium 2-ethylhexanoate, 1,5,7-triazabicyclo[4.4.0]dec-5-ene and 7-methyl-1,5,7-triazabicyclo[4.4.0] dec-5-ene.

13. A nanomicelle drug-loading system, comprising the amphiphilic block copolymer according to claim 1 and drug.

14. The nanomicelle drug-loading system according to claim 13, wherein a weight ratio of the drug and the amphiphilic block copolymer is (0.5-100):100.

15. The nanomicelle drug-loading system according to claim 13, wherein the nanomicelle drug-loading system further comprises a pharmaceutically acceptable pharmaceutical ingredient.

16. A method of preparing the nanomicelle drug-loading system according to claim 13, wherein the preparation comprises dialysis, solvent evaporation or thin-film rehydration.

17. The amphiphilic block copolymer according to claim 2, wherein in the aromatic ring, the C.sub.6-C.sub.20 aryl or R.sup.a substituted C.sub.6-C.sub.20 aryl is phenyl or naphthyl; and/or, in the aromatic ring, the C.sub.2-C.sub.20 heteroaryl or R.sup.b substituted C.sub.2-C.sub.20 heteroaryl is C.sub.3-C.sub.8 heteroaryl; and/or, the C.sub.1-C.sub.30 small molecular fragment optionally comprises a heteroatom substitution; wherein the number of heteroatom(s) is 1-4.

18. The amphiphilic block copolymer according to claim 4, wherein, in the aromatic ring, the C.sub.6-C.sub.20 aryl or the R.sup.a substituted C.sub.6-C.sub.20 aryl is C.sub.6-C.sub.10 aryl; and/or, in the aromatic ring, the C.sub.2-C.sub.20 heteroaryl or R.sup.b substituted C.sub.2-C.sub.20 heteroaryl is C.sub.2-C.sub.10 heteroaryl; and/or, in R.sup.a or R.sup.b, the C.sub.1-C.sub.6 alkyl is C.sub.1-C.sub.3 alkyl; and/or, in R.sup.a or R.sup.b, the C.sub.1-C.sub.6 alkoxy is C.sub.1-C.sub.3 alkoxy; and/or, the C.sub.1-C.sub.30 small molecular fragment is a C.sub.2-C.sub.10 small molecular fragment; and/or, the C.sub.1-C.sub.30 small molecular fragment is substituted by 1-3 aromatic rings; and/or, the C.sub.1-C.sub.30 small molecular fragment optionally comprises a heteroatom substitution; wherein the heteroatom is one or more heteroatom(s) selected from the group consisting of oxygen nitrogen, sulfur and phosphorus, wherein the number of heteroatom(s) is one or more; and/or, the hydrophobic chain segment is one chain segment selected from a polylactide chain segment or a mono-protected polylactide chain segment with the number-average molecular weight ranging from 400 to 20000; and/or, the configuration of the amino acid in the amino acid containing aromatic ring is R, S or racemic; and/or, the configuration of the amino alcohol in the amino alcohol containing aromatic ring is R, S or racemic.

19. The preparation method of the amphiphilic block copolymer according to claim 12, wherein the catalyst in step 2) is 1,8-diazabicyclo[5.4.0]undec-7-ene and/or stannous octoate.

20. The nanomicelle drug-loading system according to claim 14, wherein the weight ratio of the drug and the amphiphilic block copolymer is (1-70):100; and/or, the drug is selected from the group consisting of paclitaxel, docetaxel, cabazitaxel, 7-epipaclitaxel, t-acetylpaclitaxel, 10-deacetylpaclitaxel, 10-deacetyl-7-epipaclitaxel, 7-xylosylpaclitaxel, 10-desacetyl-7-glutarylpaclitaxel, 7-N,N-dimethylglycylpaclitaxel, 7-L-alanylpaclitaxel, larotaxel, doxorubicin, epirubicin, SN-38, irinotecan, topotecan, cyclophosphamide, ifosfamide, estramustine, mitoxantrone, amsacrine, cisplatin, carboplatin, oxaliplatin, etoposide, teniposide, vinblastine, vincristine, vinorelbine, vindesine, maytansine, harringtonine, homoharringtonine, mitomycin, bleomycin, daunorubicin, idarubicin, doxorubicin, epirubicin, gemcitabine, capecitabine, fludarabine, cladribine, bortezomib, carfilzomib, ixazomib, carmustine, fluorouracil, cytarabine, cyclosporin A, sirolimus, temsirolimus, everolimus, eribulin, trabectedin, fulvestrant, letrozole, temozolomide, raloxifene, tamoxifen, lenalidomide, ixabepilone, methotrexate, pemetrexed, enzalutamide, abiraterone, bendamustine, curcumin, resveratrol, indomethacin, huperzine A, acyclovir, allopurinol, amiodarone, azathioprine, benazepril, calcitriol, candesartan, eprosartan, carbidopa/levodopa, clarithromycin, clozapine, desmopressin acetate, diclofenac, enalapril, famotidine, felodipine, fenofibrate, fentanyl, fexofenadine, fosinopril, furosemide, glibenclamide, scopolamine, imipramine, itraconazole, levothyroxine, atorvastatin, lovastatin, meclizine, megestrol, thiopurine, metolazone, mometasone, nabumetone, omeprazole, paroxetine, propafenone, quinapril, simvastatin, sirolimus, tacrolimus, tizanidine, risperidone, olanzapine, ziprasidone, rivastigmine, naloxone, naltrexone, sirolimus, tacrolimus, carmustine, progesterone, estrogen, estradiol, levonorgestrel, norethisterone, ixabepilone, epothilone, rapamycin, plicamycin, vancomycin, amphotericin B, etoposide, doxycycline, itraconazole, fluconazole, voriconazole, posaconazole, ketoconazole, testosterone, progesterone, triamcinolone, dexamethasone, tenoxicam, piroxicam, ibuprofen, caspofungin, micafungin, olaparib, butylphthalide, combretastatin, GW6471, COX-II inhibitor, aromatase inhibitor, peptide drugs and a combination thereof.

21. The nanomicelle drug-loading system according to claim 15, wherein the pharmaceutically acceptable pharmaceutical ingredient comprises a freeze-dried excipient.

22. The nanomicelle drug-loading system according to claim 21, wherein the freeze-dried excipient is one or more excipients selected from the group consisting of lactose, mannose, sucrose, trehalose, fructose, glucose, sodium alginate and gelatin.

23. The method of claim 2 wherein the preparation method comprises thin-film rehydration.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1 is the gel permeation chromatograph of the copolymer of formula Ia, PDI=1.07;

(2) FIG. 2 is the gel permeation chromatograph of the copolymer of formula Ib, PDI=1.05;

(3) FIG. 3 is the gel permeation chromatograph of the copolymer of formula Ic, PDI=1.07;

(4) FIG. 4 is the gel permeation chromatograph of the copolymer of formula Id, PDI=1.07;

(5) FIG. 5 is the gel permeation chromatograph of the copolymer of formula Ie, PDI=1.06;

(6) FIG. 6 is the gel permeation chromatograph of the copolymer of formula Il, PDI=1.07;

(7) FIG. 7 is the H-NMR spectrum of the copolymer of formula Ia;

(8) FIG. 8 is the H-NMR spectrum of the copolymer of formula Ib;

(9) FIG. 9 is the H-NMR spectrum of the copolymer of formula Ic;

(10) FIG. 10 is the H-NMR spectrum of the copolymer of formula Id;

(11) FIG. 11 is the H-NMR spectrum of the copolymer of formula Ie;

(12) FIG. 12 is the H-NMR spectrum of the copolymer of formula Il;

(13) FIG. 13 is the transmission electron micrograph of the micelle by the copolymer of formula Ia and paclitaxel;

(14) FIG. 14 is the particle size distribution diagram of the micelle formed by the copolymer of formula Ia and paclitaxel;

(15) FIG. 15 is a graph showing the stability comparison of the micelle formed by unmodified PEG-PLA (from Samyang, Korea) and paclitaxel (weight ratio of paclitaxel and the copolymer as 20:100), and the micelle formed by the copolymer of formula Ia in the present disclosure and paclitaxel (weight ratio of paclitaxel and the copolymer as 20:100);

(16) FIG. 16 is a graph showing the stability comparison of the micelles formed by paclitaxel and the copolymer of formula Ia in different ratio;

(17) FIG. 17 is the inhibition effect graph of micellar paclitaxel from the present disclosure, micellar paclitaxel from Genexol-PM, and paclitaxel injection on the Colo-205 tumor volume;

(18) FIG. 18 is changing curve graph of micellar paclitaxel from the present disclosure, micellar paclitaxel from Genexol-PM, and paclitaxel injection on the weight of nude mice;

(19) FIG. 19 is inhibition effect graph of micellar paclitaxel from the present disclosure, micellar paclitaxel from Genexol-PM, and paclitaxel injection on the MCF-7 tumor volume;

(20) FIG. 20 is changing curve graph of micellar paclitaxel from the present disclosure, micellar paclitaxel from Genexol-PM, and paclitaxel injection on the weight of nude mice;

(21) FIG. 21 is the state diagram of micelle from unmodified PEG-PLA;

(22) FIG. 22 is the state diagram of micelle from PEG-PLA with modification at the termini of PLA;

(23) FIG. 23 is the state diagram of micelle from PLA-linker-PLA in the present disclosure.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

(24) The present disclosure provides specific embodiments, but the present disclosure is not limited thereto. Those concrete conditions and experimental methods which are not defined in the specific embodiments can follow conventional methods and conditions, or can be selected from commercial instructions.

Example 1: Preparation of the Compound of Formula VIa

(25) 4.48 g lactic acid was added to the flask, and dissolved by 100 mL dichloromethane. After imidazole (16.2 g) was added and dissolved, TBSC1 (18 g) was then added under stirring. The reaction was stirred at room temperature for 16 h and quenched by water. Workup and concentration to get crude intermediate. The crude intermediate was dissolved by 200 mL MeOH and 100 mL K2CO3 aqueous solution. The resulting solution was stirred at room ambient temperature for 3 h, extracted by EtOAc, concentrated to get the second crude intermediate. The second crude intermediate was dissolved by 50 mL dichloromethane. After addition of DCC (7 g) and NHS (5.4 g), the reaction was stirred at room temperature for 16 h. After filtration, the filtrate was concentrated to get a solid-liquid mixture crude. Purification by silica gel chromatography yielded 8.6 g compound of formula VIa.

(26) .sup.1H NMR (400 MHz, Chloroform-d) δ 4.64 (q, J=6.8 Hz, 1H), 2.95-2.71 (m, 4H), 1.63-1.49 (m, 3H), 0.90 (s, 9H), 0.12 (d, J=4.8 Hz, 6H).

Example 2: Preparation of the Compound of Formula Va

(27) 10 g polyethylene glycol monomethyl ether (number-average molecular weight of 2000) was added to the flask, dissolved by 50 mL dichloromethane. Under stirring, Boc-protected L-phenylalanine (3.6 g), EDCI (5.71 g) and DMAP (1.52 g) were sequentially added to the mixture. The reaction was stirred at room temperature for 24 h and sequentially washed with 1 N hydrochloric acid, saturated sodium hydrocarbonate. After separation and concentration to remain a small amount of dichloromethane, MTBE was used for precipitation. The solid was filtered over Buchner funnel to get 5.9 g compound of formula Va.

(28) .sup.1H NMR (400 MHz, Chloroform-d) δ7.26 (m, 5H), 4.99 (d, J=8.3 Hz, 1H), 4.59 (m, 1H), 4.32-4.17 (m, 2H), 3.62 (s, 197H), 3.36 (s, 3H), 3.08 (m, 2H), 1.40 (s, 9H).

Example 3: Preparation of the Compound of Formula IVa

(29) 8 g compound of formula Va was added to the flask, and dissolved by 15 mL dichloromethane. Under stirring, 10 mL trifluoroacetic acid was added and the solution was stirred at room temperature for 18 h. After pH was adjusted to 7-8, the mixture was extracted by dichloromethane. After separation and concentration to remain a small amount of dichloromethane, MTBE was used for precipitation. The solid was filtered over Buchner funnel to get 6 g compound of formula IVa.

(30) .sup.1H NMR (400 MHz, Chloroform-d) δ7.26 (m, 5H), 4.33-4.19 (m, 2H), 4.01-3.41 (m, 195H), 3.37 (s, 3H), 3.18-2.97 (m, 2H).

Example 4: Preparation of the Compound of Formula IIIa

(31) 1.8 g compound of formula IVa was added to the flask, and dissolved by 20 mL dichloromethane. Under stirring, compound of formula VIa (0.4 g) was added and the reaction was continued to stir for 18 h at room temperature. MTBE was used for precipitation. The solid was filtered over Buchner funnel to get 1.2 g compound of formula IIIa.

(32) .sup.1H NMR (400 MHz, Chloroform-d) δ7.26 (m, 6H), 4.87 (m, J=8.6, 5.7 Hz, 1H), 4.31-4.10 (m, 3H), 3.62 (m, 188H), 3.35 (s, 3H), 3.19-3.04 (m, 2H), 1.29 (d, J=6.7 Hz, 3H), 0.81 (s, 9H), 0.02 (d, J=14.2 Hz, 6H).

Example 5: Preparation of the Compound of Formula IIa

(33) Acetic acid (6 g) was added to the solution of KF (2 g) in 40 mL H2O. To the solution was added compound of formula IIIa (6 g). The reaction was stirred at room temperature for 24 h. After pH was adjusted to 7-8, the resulting mixture was extracted by dichloromethane. The organic phase was separated and washed by brine and precipitated by MTBE. The solid was filtered over Buchner funnel to get 4 g compound of formula IIa.

(34) .sup.1H NMR (400 MHz, Chloroform-d) δ7.26 (m, 6H), 4.89 (m, 1H), 4.41-4.08 (m, 3H), 3.63 (s, 191H), 3.36 (s, 3H), 3.27-3.03 (m, 2H), 1.31 (d, J=6.8 Hz, 3H).

Example 6: Preparation of the Compound of Formula Ia

(35) 1.65 g compound of formula IIa and DL-lactide (1.54 g) were sequentially added to the flask, and dissolved by 8 mL dichloromethane under stirring. After addition of DBU (46 mg), the reaction was stirred at room temperature for 1 h. The solution was diluted by 50 mL dichloromethane. The organic phase was sequentially washed by 1 N hydrochloric acid and brine. MTBE was used for precipitation. The solid was filtered over Buchner funnel to get 2.5 g compound of formula Ia. The molecular weight was calculated to be 3900 according to H-NMR. PDI (polydisperity index) was 1.07 according to GPC (gel permeation chromatography).

(36) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ8.54-8.37 (m, 1H), 7.35-7.14 (m, 5H), 5.48 (d, J=5.9 Hz, 1H), 5.18 (m, 22H), 5.01 (m, 1H), 4.53-4.42 (m, 1H), 4.16 (m, 3H), 3.50 (s, 189H), 3.24 (s, 3H), 3.12-2.91 (m, 2H), 1.57-1.17 (m, 76H).

Example 7: Preparation of the Compound of Formula Va′

(37) 10 g polyethylene glycol monomethyl ether (number-average molecular weight of 2000) was added to the flask, dissolved by 50 mL dichloromethane. Under stirring, Boc-protected phenylalanine (3.6 g), EDCI (5.71 g) and DMAP (1.52 g) were sequentially added to the mixture. The reaction was stirred at room temperature for 24 h and sequentially washed with 1 N hydrochloric acid, saturated sodium hydrocarbonate. After separation and concentration to remain a small amount of dichloromethane, MTBE was used for precipitation. The solid was filtered over Buchner funnel to get 9.5 g compound of formula Va′.

(38) .sup.1H NMR (400 MHz, Chloroform-d) δ7.26 (m, 5H), 4.99 (d, J=8.3 Hz, 1H), 4.59 (m, 1H), 4.32-4.17 (m, 2H), 3.62 (s, 197H), 3.36 (s, 3H), 3.08 (m, 2H), 1.40 (s, 9H).

Example 8: Preparation of the Compound of Formula IVa′

(39) 8 g compound of formula Va′ was added to the flask, and dissolved by 15 mL dichloromethane. Under stirring, 10 mL trifluoroacetic acid was added and the solution was stirred at room temperature for 18 h. After pH was adjusted to 7-8, the mixture was extracted by dichloromethane. After separation and concentration to remain a small amount of dichloromethane, MTBE was used for precipitation. The solid was filtered over Buchner funnel to get 6 g compound of formula IVa′.

(40) .sup.1H NMR (400 MHz, Chloroform-d) δ7.26 (m, 5H), 4.33-4.19 (m, 2H), 4.01-3.41 (m, 195H), 3.37 (s, 3H), 3.18-2.97 (m, 2H).

Example 9: Preparation of the Compound of Formula Ia′

(41) 1.65 g compound of formula IIa and DL-lactide (1.54 g) were sequentially added to the flask, and dissolved by 8 mL toluene under stirring. After addition of stannous octoate (46 mg), the reaction was stirred at 80° C. for 5 h. The solution was diluted by 50 mL dichloromethane. The organic phase was sequentially washed by 1 N hydrochloric acid and brine. MTBE was used for precipitation. The solid was filtered over Buchner funnel to get 2.5 g compound of formula Ia′. The molecular weight was calculated to be 3900 according to H-NMR. PDI (polydisperity index) was 1.04 according to GPC (gel permeation chromatography).

(42) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ8.54-8.37 (m, 1H), 7.35-7.14 (m, 5H), 5.48 (d, J=5.9 Hz, 1H), 5.18 (m, 22H), 5.01 (m, 1H), 4.53-4.42 (m, 1H), 4.16 (m, J=27.5, 12.4, 6.1 Hz, 3H), 3.50 (s, 189H), 3.24 (s, 3H), 3.12-2.91 (m, 2H), 1.57-1.17 (m, 76H).

Example 10: Preparation of the Compound of Formula Vb

(43) 3 g polyethylene glycol monomethyl ether (number-average molecular weight of 2000) was added to the flask, dissolved by 20 mL dichloromethane. Under stirring, the mixture was sequentially added Boc-protected 3-(2-naphthyl)-alanine (1.4 g), EDCI (1.71 g) and DMAP (0.55 g). The reaction was stirred at room temperature for 18 h and sequentially washed with 1 N hydrochloric acid, saturated sodium hydrocarbonate. MTBE was used for precipitation. The solid was filtered over Buchner funnel to get 2.5 g compound of formula Vb.

(44) .sup.1H NMR (400 MHz, Chloroform-d) δ 7.83-7.73 (m, 3H), 7.63-7.58 (m, 1H), 7.49-7.40 (m, 2H), 7.26 (d, 2H), 5.03 (d, J=8.2 Hz, 1H), 4.67 (dt, J=8.1, 5.9 Hz, 1H), 4.35-4.17 (m, 2H), 3.63 (m, 188H), 3.37 (s, 3H), 3.25 (m, 2H), 1.39 (s, 9H).

Example 11: Preparation of the Compound of Formula IVb

(45) 2.7 g compound of formula Vb was added to the flask, and dissolved by 10 mL dichloromethane. Under stirring, 8 mL trifluoroacetic acid was added and the solution was stirred at room temperature for 5 h. After pH was adjusted to 7-8, the mixture was extracted by dichloromethane. After separation and concentration to remain a small amount of dichloromethane, MTBE was used for precipitation. The solid was filtered over Buchner funnel to get 2.5 g compound of formula IVb.

(46) .sup.1H NMR (400 MHz, Chloroform-d) δ 7.78 (t, J=8.0 Hz, 3H), 7.66 (s, 1H), 7.44 (m, 2H), 7.33 (m, 1H), 4.26 (m, 2H), 3.85 (m, 1H), 3.63 (s, 191H), 3.36 (s, 3H), 3.30-3.17 (m, 1H), 3.05 (dd, J=13.5, 7.7 Hz, 1H).

Example 12: Preparation of the Compound of Formula IIIb

(47) 2.5 g compound of formula IVb was added to the flask, and dissolved by 10 mL dichloromethane. Under stirring, compound of formula VIa (0.5 g) was added and the reaction was continued to stir for 12 h at room temperature. MTBE was used for precipitation. The solid was filtered over Buchner funnel to get 2.4 g compound of formula IIIb.

(48) .sup.1H NMR (400 MHz, Chloroform-d) δ7.26 (m, 6H), 4.87 (m, J=8.6, 5.7 Hz, 1H), 4.31-4.10 (m, 3H), 3.62 (m, 188H), 3.35 (s, 3H), 3.19-3.04 (m, 2H), 1.29 (d, J=6.7 Hz, 3H), 0.81 (s, 9H), 0.02 (d, J=14.2 Hz, 6H).

Example 13: Preparation of the Compound of Formula IIb

(49) Acetic acid (2.4 g) was added to the solution of KF (0.8 g) in 20 mL H2O. To the solution was added compound of formula IIIb (2.4 g). The reaction was stirred at room temperature for 6 h. After pH was adjusted to 7-8, the resulting mixture was extracted by dichloromethane. The organic phase was separated and washed by brine and precipitated by MTBE. The solid was filtered over Buchner funnel to get 1.2 g compound of formula IIb.

(50) .sup.1H NMR (400 MHz, Chloroform-d) δ 7.84-7.70 (m, 3H), 7.60 (d, J=1.6 Hz, 1H), 7.50-7.37 (m, 2H), 7.12 (d, J=8.3 Hz, 1H), 4.97 (m, 1H), 4.33-4.12 (m, 3H), 3.63 (s, 189H), 3.36 (s, 5H), 1.31 (d, J=6.8 Hz, 3H).

Example 14: Preparation of the Compound of Formula Ib

(51) 1 g compound of formula IIb and DL-lactide (0.73 g) were sequentially added to the flask, and dissolved by 10 mL toluene under stirring. After addition of magnesium 2-ethylhexanoate (24 mg), the reaction was stirred at 90° C. for 3 h. The solution was diluted by dichloromethane. The organic phase was sequentially washed by 1 N hydrochloric acid and brine. MTBE was used for precipitation. The solid was filtered over Buchner funnel to get 1.2 g compound of formula Ib. The molecular weight was calculated to be 4200 according to H-NMR. PDI was 1.05 according to GPC.

(52) .sup.1H NMR (400 MHz, Chloroform-d) δ 7.77 (m, 3H), 7.60 (d, J=4.3 Hz, 1H), 7.44 (m, 2H), 7.26 (s, 1H), 6.69-6.50 (m, 1H), 5.28-4.82 (m, 22H), 4.30 (m, 3H), 3.63 (s, 185H), 3.37 (s, 5H), 2.74 (d, J=14.1 Hz, 1H), 1.66-1.33 (m, 67H).

Example 15: Preparation of the Compound of Formula IIIc

(53) 10 g polyethylene glycol monomethyl ether (number-average molecular weight of 2000) was added to the flask, dissolved by 50 mL dichloromethane. Under stirring, the mixture was sequentially added succinic anhydride (1 g) and DMAP (0.6 g). After stirring for 5 h, the reaction mixture was precipitated by MTBE. The solid was filtered over Buchner funnel to get 7 g compound of formula IIIc.

(54) .sup.1H NMR (400 MHz, Chloroform-d) δ 4.25 (q, J=4.1 Hz, 2H), 3.63 (m, 186H), 3.37 (s, J=2.9 Hz, 3H), 2.63 (m, 4H).

Example 16: Preparation of the Compound of Formula IIc

(55) 3.5 g compound of formula IIIc was added to the flask, and dissolved by 15 mL DMF. Under stirring, the mixture was sequentially added phenylalaninol (0.8 g), EDCI (1.9 g) and HOBT (1.34 g). After stirring at room temperature for 18 h, the organic phase was sequentially washed with 1 N hydrochloric acid and brine, then precipitated by MTBE. The solid was filtered over Buchner funnel to get 2.5 g compound of formula IIc.

(56) .sup.1H NMR (400 MHz, Chloroform-d) δ 7.26 (s, 5H), 6.24 (d, J=8.0 Hz, 1H), 4.28-4.19 (m, 2H), 4.14 (m, 1H), 3.63 (m, 195H), 3.37 (s, 3H), 3.02 (s, 1H), 2.86 (d, J=7.4 Hz, 2H), 2.79-2.56 (m, 2H), 2.50-2.38 (m, 2H).

Example 17: Preparation of the Compound of Formula Ic

(57) 1 g compound of formula IIc, D-lactide (0.42 g) and L-lactide (0.42 g) were sequentially added to the flask, and dissolved by 10 mL dichloromethane under stirring. After addition of DBU (30 mg), the reaction was stirred at 50° C. for 10 min. The organic phase was sequentially washed by 1 N hydrochloric acid and brine. MTBE was used for precipitation. The solid was filtered over Buchner funnel to get 1.2 g compound of formula Ic. The molecular weight was calculated to be 4000 according to H-NMR. PDI was 1.07 according to GPC.

(58) .sup.1H NMR (400 MHz, Chloroform-d) δ7.26 (s, 5H), 6.19-5.96 (m, 1H), 5.17 (m, 23H), 4.36 (td, 2H), 4.27-4.05 (m, 4H), 3.64 (s, 197H), 3.38 (s, 3H), 2.93-2.68 (m, 2H), 2.64 (t, J=6.9 Hz, 2H), 2.48-2.38 (m, 2H), 1.66-1.43 (m, 73H).

Example 18: Preparation of the Compound of Formula IId

(59) 2.5 g compound of formula IIIc was added to the flask, and dissolved by 15 mL DMF. Under stirring, the mixture was sequentially added 3-(2-naphthyl)-alaninol (0.8 g), EDCI (0.9 g) and HOBT (0.5 g). After stirring at room temperature for 24 h, the organic phase was sequentially washed with 1 N hydrochloric acid, saturated NaHCO.sub.3 aqueous solution and brine, then precipitated by MTBE. The solid was filtered over Buchner funnel to get 2 g compound of formula IId.

(60) .sup.1H NMR (400 MHz, Chloroform-d) δ 7.83-7.71 (m, 3H), 7.67 (d, J=1.6 Hz, 1H), 7.51-7.33 (m, 3H), 6.29 (d, J=7.9 Hz, 1H), 4.31-4.13 (m, 3H), 3.63 (s, 193H), 3.37 (s, 3H), 3.03 (d, J=7.5 Hz, 3H), 2.80-2.57 (m, 2H), 2.44 (m, 2H).

Example 19: Preparation of the Compound of Formula Id

(61) 1 g compound of formula IId, D-lactide (0.41 g) and L-lactide (0.41 g) were sequentially added to the flask, and dissolved by 10 mL dichloromethane under stirring. After addition of DBU (28 mg), the reaction was stirred at 25° C. for 1 h. The organic phase was sequentially washed by 1 N hydrochloric acid and brine. MTBE was used for precipitation. The solid was filtered over Buchner funnel to get 1.2 g compound of formula Id. The molecular weight was calculated to be 4000 according to H-NMR. PDI was 1.07 according to GPC.

(62) .sup.1H NMR (400 MHz, Chloroform-d) δ 7.79 (t, J=8.7 Hz, 3H), 7.63 (s, 1H), 7.45 (m, 2H), 7.37-7.30 (m, 1H), 6.32-6.08 (m, 1H), 5.31-5.06 (m, 23H), 4.52 (s, 1H), 4.35 (p, J=6.8 Hz, 1H), 4.15 (ddd, J=17.4, 6.6, 5.6 Hz, 4H), 3.63 (d, J=3.0 Hz, 200H), 3.37 (s, 3H), 3.11-2.89 (m, 2H), 2.75 (s, 1H), 2.64 (t, J=6.9 Hz, 2H), 2.44 (dd, J=6.8, 3.1 Hz, 2H), 1.67-1.43 (m, 73H).

Example 20: Preparation of the Compound of Formula IIIe

(63) 8 g polyethylene glycol monomethyl ether (number-average molecular weight of 2000) was added to the flask, dissolved by 100 mL dichloromethane. Under stirring, the mixture was sequentially added p-formylbenzoic acid (2.5 g), DCC (6.56 g) and DMAP (2.18 g). After stirring at room temperature for 10 h, the reaction mixture was precipitated by MTBE. The solid was filtered over Buchner funnel to get 5.5 g compound of formula IIIe.

(64) .sup.1H NMR (400 MHz, Chloroform-d) δ 10.08 (s, 1H), 8.26-8.14 (m, 2H), 8.00-7.88 (m, 2H), 4.52-4.45 (m, 2H), 3.62 (s, 189H), 3.36 (s, 3H).

Example 21: Preparation of the Compound of Formula IIe

(65) 2.5 g compound of formula IIIe was added to the flask, dissolved by 50 mL ethanol. Under stirring, the mixture was added sodium borohydride (100 mg). After stirring for 1 h, the reaction mixture was sequentially washed with 1 N hydrochloric acid and brine. The organic phase was precipitated by MTBE. The solid was filtered and dried under vacuum at room temperature to get 0.8 g compound of formula IIe.

(66) .sup.1H NMR (400 MHz, Chloroform-d) δ 8.02 (d, J=8.2 Hz, 2H), 7.42 (d, J=8.0 Hz, 2H), 4.74 (d, J=5.7 Hz, 2H), 4.45 (m, 2H), 3.62 (m, 203H), 3.36 (s, 3H), 2.65 (t, J=6.1 Hz, 1H).

Example 22: Preparation of the Compound of Formula Ie

(67) 0.8 g compound of formula IIe, DL-lactide (0.7 g) were sequentially added to the flask, and dissolved by 10 mL dichloromethane. After addition of DBU (16 mg), the reaction was stirred at 10° C. for 30 min. The organic phase was sequentially washed by 1 N hydrochloric acid and brine. MTBE was used for precipitation. The solid was filtered and dried under vacuum at room temperature to get 0.8 g compound of formula Ie. The molecular weight was calculated to be 3800 according to H-NMR. PDI was 1.06 according to GPC.

(68) .sup.1H NMR (400 MHz, Chloroform-d) δ 8.14-7.95 (m, 2H), 7.37 (d, J=8.0 Hz, 2H), 5.31-5.09 (m, 20H), 4.53-4.41 (m, 2H), 4.34 (p, J=6.8 Hz, 1H), 3.63 (m, 200H), 3.37 (s, 3H), 2.73 (d, J=14.5 Hz, 1H), 1.68-1.41 (m, 60H).

Example 23: Preparation of the Compound of Formula if

(69) According to the synthetic route of the compound of formula Ia, using polyethylene glycol monomethyl ether (number-average molecular weight of 4000), racemic tryptophan, DL-lactide as starting material, through introduction of linker to initiate polymerization, to get compound of formula If. The molecular weight was calculated to be 7900 according to H-NMR. PDI was 1.07 according to GPC.

(70) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ10.5 (s, 1H), 7.62 (d, 1H), 7.30 (d, 1H), 7.18 (s, 1H), 7.12 (m, 2H), 5.48 (d, J=5.9 Hz, 1H), 5.18 (m, 54H), 5.01 (m, 1H), 4.53-4.42 (m, 1H), 4.16 (m, 3H), 3.50 (s, 364H), 3.24 (s, 3H), 3.30-3.12 (m, 2H), 1.57-1.17 (m, 160H).

Example 24: Preparation of the Compound of Formula Ig

(71) According to the synthetic route of the compound of formula Ia, using polyethylene glycol monomethyl ether (number-average molecular weight of 5000), tyrosine, DL-lactide as starting material, through introduction of linker to initiate polymerization, to get compound of formula Ig. The molecular weight was calculated to be 10000 according to H-NMR. PDI was 1.09 according to GPC.

(72) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ7.15 (d, 2H), 6.8 (d, 2H), 5.48 (d, J=5.9 Hz, 1H), 5.33 (s, 1H), 5.18 (m, 68H), 5.01 (m, 1H), 4.53-4.42 (m, 1H), 4.16 (m, 3H), 3.50 (s, 455H), 3.24 (s, 3H), 3.42-3.20 (m, 2H), 1.57-1.17 (m, 208H).

Example 25: Preparation of the Compound of Formula Ih

(73) According to the synthetic route of the compound of formula Ia, using polyethylene glycol monomethyl ether (number-average molecular weight of 2000), histidine, DL-lactide as starting material, through introduction of linker to initiate polymerization, to get compound of formula Ih. The molecular weight was calculated to be 5000 according to H-NMR. PDI was 1.05 according to GPC.

(74) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ12.0 (s, 1H), 8.77 (s, 1H), 7.63 (s, 1H), 5.48 (d, J=5.9 Hz, 1H), 5.18 (m, 42H), 5.01 (m, 1H), 4.53-4.42 (m, 1H), 4.16 (m, 3H), 3.50 (s, 180H), 3.24 (s, 3H), 3.82-3.56 (m, 2H), 1.57-1.17 (m, 126H).

Example 26: Preparation of the Compound of Formula Ii

(75) According to the synthetic route of the compound of formula Ia, using polyethylene glycol monobenzyl ether (number-average molecular weight of 2000), phenylalanine, DL-lactide as starting material, through introduction of linker to initiate polymerization, to get compound of formula Ih. The molecular weight was calculated to be 4000 according to H-NMR. PDI was 1.06 according to GPC.

(76) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ8.54-8.37 (m, 1H), 7.45 (d, 2H), 7.38-7.14 (m, 8H), 5.48 (d, J=5.9 Hz, 1H), 5.18 (m, 23H), 5.01 (m, 1H), 4.82 (s, 2H), 4.53-4.42 (m, 1H), 4.16 (m, J=27.5, 12.4, 6.1 Hz, 3H), 3.50 (s, 189H), 3.12-2.91 (m, 2H), 1.57-1.17 (m, 68H).

Example 27: Preparation of the Compound of Formula Ij

(77) According to the synthetic route of the compound of formula Ia, using polyethylene glycol monobenzyl ether (number-average molecular weight of 10000), phenylalanine, glycine, DL-lactide as starting material, through introduction of linker to initiate polymerization, to get compound of formula Ij. The molecular weight was calculated to be 20000 according to H-NMR. PDI was 1.08 according to GPC.

(78) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ8.54-8.37 (m, 1H), 7.35-7.14 (m, 5H), 5.48 (d, J=5.9 Hz, 1H), 5.18 (m, 138H), 5.01 (m, 1H), 4.53-4.42 (m, 1H), 4.16 (m, J=27.5, 12.4, 6.1 Hz, 3H), 4.03 (s, 2H), 3.50 (s, 910H), 3.24 (s, 3H), 3.12-2.91 (m, 2H), 1.57-1.17 (m, 421H).

Example 28: Preparation of the Compound of Formula Ik

(79) According to the synthetic route of the compound of formula Ia, using polyethylene glycol monomethyl ether (number-average molecular weight of 6000), phenylalanine, glycine, D-tyrosine, DL-lactide as starting material, through introduction of linker to initiate polymerization, to get compound of formula Ik. The molecular weight was calculated to be 14000 according to H-NMR. PDI was 1.07 according to GPC.

(80) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ8.54-8.37 (m, 1H), 7.35-7.14 (m, 5H), 7.10 (d, 2H), 6.75 (d, 2H), 5.48 (d, J=5.9 Hz, 1H), 5.18 (m, 108H), 5.01 (m, 1H), 4.53-4.42 (m, 1H), 4.16 (m, 3H), 3.50 (s, 545H), 3.24 (s, 3H), 3.08-2.88 (m, 2H), 3.12-2.91 (m, 2H), 1.57-1.17 (m, 320H).

Example 29: Preparation of the Compound of Formula Il

(81) According to the synthetic route of the compound of formula Ia, using polyethylene glycol monomethyl ether (number-average molecular weight of 2000), phenylalanine, DL-lactide as starting material, through introduction of linker to initiate polymerization, and protecting the termini of PLA by acetyl group to get compound of formula Il. The molecular weight was calculated to be 7000 according to H-NMR. PDI was 1.06 according to GPC.

(82) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ8.54-8.37 (m, 1H), 7.35-7.14 (m, 5H), 5.48 (d, J=5.9 Hz, 1H), 5.18 (m, 66H), 5.01 (m, 1H), 4.53-4.42 (m, 1H), 4.16 (m, J=27.5, 12.4, 6.1 Hz, 3H), 3.50 (s, 189H), 3.24 (s, 3H), 3.12-2.91 (m, 2H), 2.03 (s, 3H), 1.57-1.17 (m, 200H).

Example 30: Preparation of the Compound of Formula Im

(83) According to the synthetic route of the compound of formula Ia, using polyethylene glycol monomethyl ether (number-average molecular weight of 2000), phenylalanine, DL-lactide as starting material, through introduction of linker to initiate polymerization, and protecting the termini of PLA by benzoyl group to get compound of formula Im. The molecular weight was calculated to be 4500 according to H-NMR. PDI was 1.06 according to GPC.

(84) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ8.54-8.37 (m, 1H), 8.00 (d, 2H), 7.63 (d, 1H), 7.58 (dd, 2H), 7.35-7.14 (m, 5H), 5.48 (d, J=5.9 Hz, 1H), 5.18 (m, 30H), 5.01 (m, 1H), 4.53-4.42 (m, 3H), 4.16 (m, J=27.5, 12.4, 6.1 Hz, 3H), 3.50 (s, 181H), 3.24 (s, 3H), 3.12-2.91 (m, 2H), 2.56 (t, 2H), 1.57-1.17 (m, 91H).

Example 31: Preparation of the Compound of Formula in

(85) According to the synthetic route of the compound of formula Ia, using polyethylene glycol monomethyl ether (number-average molecular weight of 2000), phenylalanine, DL-lactide as starting material, through introduction of linker to initiate polymerization, and protecting the termini of PLA by Boc-protected phenylalanine to get compound of formula In. The molecular weight was calculated to be 5000 according to H-NMR. PDI was 1.04 according to GPC.

(86) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ8.54-8.37 (m, 1H), 7.35-7.14 (m, 10H), 5.48 (d, J=5.9 Hz, 1H), 5.18 (m, 37H), 5.01 (m, 2H), 4.53-4.42 (m, 3H), 4.16 (m, J=27.5, 12.4, 6.1 Hz, 3H), 3.50 (s, 180H), 3.24 (s, 3H), 3.12-2.91 (m, 4H), 2.56 (t, 2H), 1.57-1.17 (m, 112H).

Example 32: Preparation of the Compound of Formula Io

(87) According to the synthetic route of the compound of formula Ia, using polyethylene glycol monomethyl ether (number-average molecular weight of 2000), benzoyl-protected lysine, DL-lactide as starting material, through introduction of linker to initiate polymerization, to get compound of formula Io. The molecular weight was calculated to be 4000 according to H-NMR. PDI was 1.08 according to GPC.

(88) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ8.50 (m, 1H), 8.05 (d, 2H), 7.72 (d, 1H), 7.58 (dd, 2H), 5.48 (d, J=5.9 Hz, 1H), 5.18 (m, 22H), 5.01 (m, 1H), 4.53-4.42 (m, 1H), 4.16 (m, 3H), 3.50 (s, 182H), 3.32 (t, 2H), 3.24 (s, 3H), 3.12-2.91 (m, 2H), 1.78 (m, 2H), 1.57-1.17 (m, 78H).

Example 33: Preparation of the Compound of Formula Ip

(89) According to the synthetic route of the compound of formula Ia, using polyethylene glycol monomethyl ether (number-average molecular weight of 2000), dithiodipropionic acid, phenylalanine, DL-lactide as starting material, through introduction of linker to initiate polymerization, to get compound of formula Ip. The molecular weight was calculated to be 4300 according to H-NMR. PDI was 1.09 according to GPC.

(90) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ8.54-8.37 (m, 1H), 7.35-7.14 (m, 5H), 5.48 (d, J=5.9 Hz, 1H), 5.18 (m, 25H), 5.01 (m, 1H), 4.53-4.42 (m, 1H), 4.16 (m, 3H), 3.50 (s, 180H), 3.24 (s, 3H), 3.12-2.91 (m, 4H), 2.80 (t, 2H), 2.58 (t, 2H), 2.44 (t, 2H), 1.57-1.17 (m, 76H).

Example 34: Preparation of the Compound of Formula Iq

(91) According to the synthetic route of the compound of formula Ia, using polyethylene glycol monomethyl ether (number-average molecular weight of 2000), phthalimide, Boc-protected phenylalanine, DL-lactide as starting material, through introduction of linker to initiate polymerization, to get compound of formula Iq. The molecular weight was calculated to be 5000 according to H-NMR. PDI was 1.07 according to GPC.

(92) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ8.54-8.37 (m, 1H), 7.35-7.14 (m, 5H), 5.48 (d, J=5.9 Hz, 1H), 5.18 (m, 39H), 5.01 (m, 1H), 4.53-4.42 (m, 3H), 4.16 (m, J=27.5, 12.4, 6.1 Hz, 3H), 3.57 (t, 2H), 3.50 (s, 180H), 3.24 (s, 3H), 3.12-2.91 (m, 2H), 2.64 (t, 2H), 2.56 (t, 2H), 1.57-1.17 (m, 118H).

Example 35: Preparation of the Compound of Formula Ir

(93) According to the synthetic route of the compound of formula Ia, using polyethylene glycol monomethyl ether (number-average molecular weight of 2000), thioacetic acid, Boc-protected phenylalanine, DL-lactide as starting material, through introduction of linker to initiate polymerization, to get compound of formula Ir. The molecular weight was calculated to be 6000 according to H-NMR. PDI was 1.07 according to GPC.

(94) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ8.54-8.37 (m, 1H), 7.35-7.14 (m, 5H), 5.48 (d, J=5.9 Hz, 1H), 5.18 (m, 53H), 5.01 (m, 1H), 4.53-4.42 (m, 3H), 4.16 (m, J=27.5, 12.4, 6.1 Hz, 3H), 3.50 (s, 180H), 3.28 (t, 2H), 3.24 (s, 3H), 3.12-2.91 (m, 2H), 2.88 (t, 2H), 2.56 (t, 2H), 1.57-1.17 (m, 160H).

Example 36: Preparation of the Compound of Formula is

(95) According to the synthetic route of the compound of formula Ia, using polyethylene glycol monomethyl ether (number-average molecular weight of 4000), butyrolactone, Boc-protected phenylalanine, DL-lactide as starting material, through introduction of linker to initiate polymerization, to get compound of formula Is. The molecular weight was calculated to be 8000 according to H-NMR. PDI was 1.07 according to GPC.

(96) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ8.54-8.37 (m, 1H), 7.35-7.14 (m, 5H), 5.48 (d, J=5.9 Hz, 1H), 5.18 (m, 54H), 5.01 (m, 1H), 4.53-4.42 (m, 3H), 4.16 (m, J=27.5, 12.4, 6.1 Hz, 3H), 3.50 (s, 364H), 3.24 (s, 3H), 3.12-2.91 (m, 2H), 2.56 (t, 2H), 1.57-1.17 (m, 162H).

Example 37: Preparation of the Compound of Formula it

(97) According to the synthetic route of the compound of formula Ia′, using polyethylene glycol monomethyl ether (number-average molecular weight of 5000), Boc-protected phenylalanine, DL-lactide as starting material, through introduction of linker to initiate polymerization, to get compound of formula It. The molecular weight was calculated to be 9000 according to H-NMR. PDI was 1.07 according to GPC.

(98) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ8.54-8.37 (m, 1H), 7.35-7.14 (m, 5H), 5.48 (d, J=5.9 Hz, 1H), 5.18 (m, 54H), 5.01 (m, 1H), 4.53-4.42 (m, 1H), 4.16 (m, J=27.5, 12.4, 6.1 Hz, 3H), 3.50 (s, 454H), 3.24 (s, 3H), 3.12-2.91 (m, 2H), 1.57-1.17 (m, 160H).

Example 38: Preparation of the Compound of Formula Iu

(99) According to the synthetic route of the compound of formula Ia′, using polyethylene glycol monomethyl ether (number-average molecular weight of 3000), Boc-protected D-phenylalanine, DL-lactide as starting material, through introduction of linker to initiate polymerization, to get compound of formula It. The molecular weight was calculated to be 8000 according to H-NMR. PDI was 1.07 according to GPC.

(100) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ8.54-8.37 (m, 1H), 7.35-7.14 (m, 5H), 5.48 (d, J=5.9 Hz, 1H), 5.18 (m, 68H), 5.01 (m, 1H), 4.53-4.42 (m, 1H), 4.16 (m, J=27.5, 12.4, 6.1 Hz, 3H), 3.50 (s, 274H), 3.24 (s, 3H), 3.12-2.91 (m, 2H), 1.57-1.17 (m, 208H).

Example 39: Preparation of the Compound of Formula Iv

(101) According to the synthetic route of the compound of formula Ia′, using polyethylene glycol monomethyl ether (number-average molecular weight of 10000), Boc-protected L-tryptophan, DL-lactide as starting material, through introduction of linker to initiate polymerization, to get compound of formula Iv. The molecular weight was calculated to be 30000 according to H-NMR. PDI was 1.07 according to GPC.

(102) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ10.5 (s, 1H), 7.62 (d, 1H), 7.30 (d, 1H), 7.18 (s, 1H), 7.12 (m, 2H), 5.48 (d, J=5.9 Hz, 1H), 5.18 (m, 278H), 5.01 (m, 1H), 4.53-4.42 (m, 1H), 4.16 (m, 3H), 3.50 (s, 908H), 3.24 (s, 3H), 3.30-3.12 (m, 2H), 1.57-1.17 (m, 840H).

Example 40: Preparation of the Compound of Formula Iw

(103) According to the synthetic route of the compound of formula Ia′, using polyethylene glycol monomethyl ether (number-average molecular weight of 20000), Fmoc-protected L-phenylalanine, glycine, DL-lactide as starting material, through introduction of linker to initiate polymerization, to get compound of formula It. The molecular weight was calculated to be 40000 according to H-NMR. PDI was 1.07 according to GPC.

(104) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ8.54-8.37 (m, 1H), 7.35-7.14 (m, 5H), 5.48 (d, J=5.9 Hz, 1H), 5.18 (m, 275H), 5.01 (m, 1H), 4.53-4.42 (m, 1H), 4.16 (m, J=27.5, 12.4, 6.1 Hz, 3H), 3.50 (s, 1818H), 3.24 (s, 3H), 3.12-2.91 (m, 2H), 1.57-1.17 (m, 842H).

Example 41: Preparation of the Compound of Formula Ix

(105) According to the synthetic route of the compound of formula Ia′, using polyethylene glycol monomethyl ether (number-average molecular weight of 2000) (1 g), Boc-protected L-phenylalanine (132 mg), DL-lactide (970 mg) as starting material, through introduction of linker to initiate polymerization, to get compound of formula Ix. The molecular weight was calculated to be 4200 according to H-NMR. PDI was 1.07 according to GPC.

(106) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ8.54-8.37 (m, 1H), 7.35-7.14 (m, 5H), 5.48 (d, J=5.9 Hz, 1H), 5.18 (m, 26H), 5.01 (m, 1H), 4.53-4.42 (m, 1H), 4.16 (m, J=27.5, 12.4, 6.1 Hz, 3H), 3.50 (s, 189H), 3.24 (s, 3H), 3.12-2.91 (m, 2H), 1.57-1.17 (m, 79H).

Example 42: Preparation of the Compound of Formula Iy

(107) According to the synthetic route of the compound of formula Ia, using polyethylene glycol monomethyl ether (number-average molecular weight of 2000) (1 g), Boc-protected L-phenylalanine (132 mg), DL-lactide (1.22 g) as starting material, through introduction of linker to initiate polymerization, to get compound of formula Iy. The molecular weight was calculated to be 4700 according to H-NMR. PDI was 1.07 according to GPC.

(108) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ8.54-8.37 (m, 1H), 7.35-7.14 (m, 5H), 5.48 (d, J=5.9 Hz, 1H), 5.18 (m, 33H), 5.01 (m, 1H), 4.53-4.42 (m, 1H), 4.16 (m, J=27.5, 12.4, 6.1 Hz, 3H), 3.50 (s, 189H), 3.24 (s, 3H), 3.12-2.91 (m, 2H), 1.57-1.17 (m, 99H).

Example 43: Preparation of the Compound of Formula Iz

(109) According to the synthetic route of the compound of formula Ia, using polyethylene glycol monomethyl ether (number-average molecular weight of 2000) (1 g), Boc-protected L-phenylalanine (132 mg), DL-lactide (2.73 g) as starting material, through introduction of linker to initiate polymerization, to get compound of formula Iz. The molecular weight was calculated to be 5000 according to H-NMR. PDI was 1.07 according to GPC.

(110) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ8.54-8.37 (m, 1H), 7.35-7.14 (m, 5H), 5.48 (d, J=5.9 Hz, 1H), 5.18 (m, 37H), 5.01 (m, 1H), 4.53-4.42 (m, 1H), 4.16 (m, J=27.5, 12.4, 6.1 Hz, 3H), 3.50 (s, 189H), 3.24 (s, 3H), 3.12-2.91 (m, 2H), 1.57-1.17 (m, 110H).

Example 44: Preparation of the Compound of Formula Iaa

(111) According to the synthetic route of the compound of formula Ia, using polyethylene glycol monomethyl ether (number-average molecular weight of 2000) (1 g), Boc-protected L-phenylalanine (132 mg), DL-lactide (1.47 g) as starting material, through introduction of linker to initiate polymerization, to get compound of formula Iaa. The molecular weight was calculated to be 5200 according to H-NMR. PDI was 1.07 according to GPC.

(112) .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ8.54-8.37 (m, 1H), 7.35-7.14 (m, 5H), 5.48 (d, J=5.9 Hz, 1H), 5.18 (m, 40H), 5.01 (m, 1H), 4.53-4.42 (m, 1H), 4.16 (m, J=27.5, 12.4, 6.1 Hz, 3H), 3.50 (s, 189H), 3.24 (s, 3H), 3.12-2.91 (m, 2H), 1.57-1.17 (m, 123H).

Example 45: Preparation of the Lyophilized Polymeric Micellar Formulation of Paclitaxel

(113) 500 mg amphiphilic block copolymer of formula Ia and 100 mg paclitaxel were added into 500 mL flask, and dissolved by 100 mL acetonitrile. The solution was placed in the shaker and shaked for 30 min at room temperature. A transparent thin film was formed around the wall of the flask after evaporation of acetonitrile. 150 mL pure water at 30° C. was immediately introduced to this flask, and a homogeneous solution with obvious blue opalescence was formed upon shaking. The solution was filtered over 0.22 μm filtration membrance, and freeze-dried into a white powder in the lyophilizer. The content of paclitaxel in the solid was 16% as detected by HPLC.

(114) The lyophilized powder was redissolved by physiological saline to a solution with the concentration of 5 mg/mL. The solution was stable for more than 72 h at room temperature.

Example 46: Preparation of the Lyophilized Polymeric Micellar Formulation of Paclitaxel

(115) 500 mg amphiphilic block copolymer of formula Ia and 150 mg paclitaxel were added into 500 mL flask, and dissolved by 100 mL acetonitrile. The solution was placed in the shaker and shaked for 30 min at room temperature. A transparent thin film was formed around the wall of the flask after evaporation of acetonitrile. 150 mL pure water at 30° C. was immediately introduced to this flask, and a homogeneous solution with obvious blue opalescence was formed upon shaking. The solution was filtered over 0.22 μm filtration membrance, and freeze-dried into a white powder in the lyophilizer. The content of paclitaxel in the solid was 23% as detected by HPLC.

(116) The lyophilized powder was redissolved by physiological saline to a solution with the concentration of 5 mg/mL. The solution was stable for more than 72 h at room temperature.

Example 47: Preparation of the Lyophilized Polymeric Micellar Formulation of Paclitaxel

(117) 500 mg amphiphilic block copolymer of formula Ia and 200 mg paclitaxel were added into 500 mL flask, and dissolved by 100 mL acetonitrile. The solution was placed in the shaker and shaked for 2 h at room temperature. A transparent thin film was formed around the wall of the flask after evaporation of acetonitrile. 150 mL pure water at 60° C. was immediately introduced to this flask, and a homogeneous solution with obvious blue opalescence was formed upon shaking. The solution was filtered over 0.22 μm filtration membrance, and freeze-dried into a white powder in the lyophilizer. The content of paclitaxel in the solid was 28% as detected by HPLC. The result for particle size measurement was shown in FIG. 14. The average particle size was 20.2 nm after taking the average of three tests. The transmission electron micrograph was shown in FIG. 13.

(118) The lyophilized powder was redissolved by physiological saline to a solution with the concentration of 5 mg/mL. The solution was stable for more than 72 h at room temperature.

Example 48: Preparation of the Lyophilized Polymeric Micellar Formulation of Paclitaxel

(119) 500 mg amphiphilic block copolymer of formula Ia and 300 mg paclitaxel were added into 500 mL flask, and dissolved by 100 mL acetonitrile. The solution was placed in the shaker and shaked for 30 min at room temperature. A transparent thin film was formed around the wall of the flask after evaporation of acetonitrile. 150 mL pure water at 30° C. was immediately introduced to this flask, and a homogeneous solution with obvious blue opalescence was formed upon shaking. The solution was filtered over 0.22 μm filtration membrance, and freeze-dried into a white powder in the lyophilizer. The content of paclitaxel in the solid was 37% as detected by HPLC.

(120) The lyophilized powder was redissolved by physiological saline to a solution with the concentration of 5 mg/mL. The solution was stable for more than 72 h at room temperature.

Example 49: Preparation of the Lyophilized Polymeric Micellar Formulation of Paclitaxel

(121) 500 mg amphiphilic block copolymer of formula Ib and 300 mg paclitaxel were added into 500 mL flask, and dissolved by 100 mL acetonitrile. The solution was placed in the shaker and shaked for 1 h at room temperature. A transparent thin film was formed around the wall of the flask after evaporation of acetonitrile. 150 mL pure water at 50° C. was immediately introduced to this flask, and a homogeneous solution with obvious blue opalescence was formed upon shaking. The solution was filtered over 0.22 μm filtration membrance, and freeze-dried into a white powder in the lyophilizer. The content of paclitaxel in the solid was 37% as detected by HPLC.

(122) The lyophilized powder was redissolved by physiological saline to a solution with the concentration of 5 mg/mL. The solution was stable for more than 72 h at room temperature.

Example 50: Preparation of the Lyophilized Polymeric Micellar Formulation of Paclitaxel

(123) 500 mg amphiphilic block copolymer of formula Ib and 50 mg paclitaxel were added into 500 mL flask, and dissolved by 100 mL acetonitrile. The solution was placed in the shaker and shaked for 1 h at room temperature. A transparent thin film was formed around the wall of the flask after evaporation of acetonitrile. 150 mL pure water at 50° C. was immediately introduced to this flask, and a homogeneous solution with obvious blue opalescence was formed upon shaking. The solution was filtered over 0.22 μm filtration membrance, and freeze-dried into a white powder in the lyophilizer. The content of paclitaxel in the solid was 9% as detected by HPLC.

(124) The lyophilized powder was redissolved by physiological saline to a solution with the concentration of 3 mg/mL. The solution was stable for more than 72 h at room temperature.

Example 51: Preparation of the Lyophilized Polymeric Micellar Formulation of Paclitaxel

(125) 500 mg amphiphilic block copolymer of formula Ic and 150 mg paclitaxel were added into 500 mL flask, and dissolved by 100 mL acetonitrile. The solution was placed in the shaker and shaked for 30 min at room temperature. A transparent thin film was formed around the wall of the flask after evaporation of acetonitrile. 150 mL pure water at 30° C. was immediately introduced to this flask, and a homogeneous solution with obvious blue opalescence was formed upon shaking. The solution was filtered over 0.22 μm filtration membrance, and freeze-dried into a white powder in the lyophilizer. The content of paclitaxel in the solid was 23% as detected by HPLC.

(126) The lyophilized powder was redissolved by physiological saline to a solution with the concentration of 5 mg/mL. The solution was stable for more than 72 h at room temperature.

Example 52: Preparation of the Lyophilized Polymeric Micellar Formulation of Docetaxel

(127) 200 mg amphiphilic block copolymer of formula Ib and 80 mg docetaxel were added into 250 mL flask, and dissolved by 50 mL acetonitrile. The solution was placed in the shaker and shaked for 2 h at room temperature. A transparent thin film was formed around the wall of the flask after evaporation of acetonitrile. 50 mL pure water at 20° C. was immediately introduced to this flask, and a homogeneous solution with obvious blue opalescence was formed upon shaking. The solution was filtered over 0.22 μm filtration membrance, and freeze-dried into a white powder in the lyophilizer. The content of docetaxel in the solid was 28% as detected by HPLC.

(128) The lyophilized powder was redissolved by physiological saline to a solution with the concentration of 5 mg/mL. The solution was stable for more than 48 h at room temperature.

Example 53: Preparation of the Lyophilized Polymeric Micellar Formulation of Eribulin

(129) 200 mg amphiphilic block copolymer of formula Ic and 40 mg eribulin were added into 250 mL flask, and dissolved by 50 mL acetonitrile. The solution was placed in the shaker and shaked for 2 h at room temperature. A transparent thin film was formed around the wall of the flask after evaporation of acetonitrile. 50 mL pure water at 40° C. was immediately introduced to this flask, and a homogeneous solution with obvious blue opalescence was formed upon shaking. The solution was filtered over 0.22 μm filtration membrance, and freeze-dried into a white powder in the lyophilizer. The content of eribulin in the solid was 17% as detected by HPLC.

(130) The lyophilized powder was redissolved by physiological saline to a solution with the concentration of 5 mg/mL. The solution was stable for more than 48 h at room temperature.

Example 54: Preparation of the Lyophilized Polymeric Micellar Formulation of Irinotecan

(131) 200 mg amphiphilic block copolymer of formula Id and 40 mg irinotecan were added into 250 mL flask, and dissolved by 50 mL acetonitrile. The solution was placed in the shaker and shaked for 2 h at room temperature. A transparent thin film was formed around the wall of the flask after evaporation of acetonitrile. 50 mL pure water at 60° C. was immediately introduced to this flask, and a homogeneous solution with obvious blue opalescence was formed upon shaking. The solution was filtered over 0.22 μm filtration membrance, and freeze-dried into a white powder in the lyophilizer. The content of irinotecan in the solid was 16% as detected by HPLC.

(132) The lyophilized powder was redissolved by physiological saline to a solution with the concentration of 3 mg/mL. The solution was stable for more than 24 h at room temperature.

Example 55: Preparation of the Lyophilized Polymeric Micellar Formulation of SN-38

(133) 200 mg amphiphilic block copolymer of formula Ih and 20 mg SN-38 were added into 250 mL flask, and dissolved by 50 mL acetonitrile. The solution was placed in the shaker and shaked for 2 h at room temperature. A transparent thin film was formed around the wall of the flask after evaporation of acetonitrile. 50 mL pure water at 40° C. was immediately introduced to this flask, and a homogeneous solution with obvious blue opalescence was formed upon shaking. The solution was filtered over 0.22 μm filtration membrance, and freeze-dried into a white powder in the lyophilizer. The content of SN-38 in the solid was 9% as detected by HPLC.

(134) The lyophilized powder was redissolved by physiological saline to a solution with the concentration of 1.5 mg/mL. The solution was stable for more than 24 h at room temperature.

Example 56: Preparation of the Lyophilized Polymeric Micellar Formulation of Fulvestrant

(135) 200 mg amphiphilic block copolymer of formula Ih and 70 mg fulvestrant were added into 250 mL flask, and dissolved by 50 mL acetonitrile. The solution was placed in the shaker and shaked for 2 h at room temperature. A transparent thin film was formed around the wall of the flask after evaporation of acetonitrile. 50 mL pure water at 40° C. was immediately introduced to this flask, and a homogeneous solution with obvious blue opalescence was formed upon shaking. The solution was filtered over 0.22 μm filtration membrance, and freeze-dried into a white powder in the lyophilizer. The content of fulvestrant in the solid was 25% as detected by HPLC.

(136) The lyophilized powder was redissolved by physiological saline to a solution with the concentration of 2 mg/mL. The solution was stable for more than 72 h at room temperature.

Example 57: Preparation of the Lyophilized Polymeric Micellar Formulation of Bortezomib

(137) 200 mg amphiphilic block copolymer of formula Iu and 70 mg bortezomib were added into 250 mL flask, and dissolved by 50 mL dichloromethane. The solution was placed in the shaker and shaked for 2 h at room temperature. A transparent thin film was formed around the wall of the flask after evaporation of dichloromethane. 50 mL pure water at 40° C. was immediately introduced to this flask, and a homogeneous solution with obvious blue opalescence was formed upon shaking. The solution was filtered over 0.22 μm filtration membrance, and freeze-dried into a white powder in the lyophilizer. The content of bortezomib in the solid was 25% as detected by HPLC.

(138) The lyophilized powder was redissolved by physiological saline to a solution with the concentration of 2 mg/mL. The solution was stable for more than 72 h at room temperature.

Example 58: Preparation of the Lyophilized Polymeric Micellar Formulation of GW6471

(139) 200 mg amphiphilic block copolymer of formula Is and 50 mg GW6471 were added into 250 mL flask, and dissolved by 50 mL acetone. The solution was placed in the shaker and shaked for 2 h at room temperature. A transparent thin film was formed around the wall of the flask after evaporation of acetone. 50 mL pure water at 40° C. was immediately introduced to this flask, and a homogeneous solution with obvious blue opalescence was formed upon shaking. The solution was filtered over 0.22 μm filtration membrance, and freeze-dried into a white powder in the lyophilizer. The content of GW6471 in the solid was 20% as detected by HPLC.

(140) The lyophilized powder was redissolved by physiological saline to a solution with the concentration of 2 mg/mL. The solution was stable for more than 72 h at room temperature.

Example 59: Preparation of the Lyophilized Polymeric Micellar Formulation of Voriconazole

(141) 200 mg amphiphilic block copolymer of formula Ix and 70 mg voriconazole were added into 250 mL flask, and dissolved by 50 mL acetonitrile. The solution was placed in the shaker and shaked for 2 h at room temperature. A transparent thin film was formed around the wall of the flask after evaporation of acetonitrile. 50 mL pure water at 40° C. was immediately introduced to this flask, and a homogeneous solution with obvious blue opalescence was formed upon shaking. The solution was filtered over 0.22 μm filtration membrance, and freeze-dried into a white powder in the lyophilizer. The content of voriconazole in the solid was 25% as detected by HPLC.

(142) The lyophilized powder was redissolved by physiological saline to a solution with the concentration of 2 mg/mL. The solution was stable for more than 72 h at room temperature.

Example 60: Preparation of the Lyophilized Polymeric Micellar Formulation of Dexamethasone

(143) 200 mg amphiphilic block copolymer of formula Iy and 90 mg dexamethasone were added into 250 mL flask, and dissolved by 50 mL acetonitrile. The solution was placed in the shaker and shaked for 2 h at room temperature. A transparent thin film was formed around the wall of the flask after evaporation of acetonitrile. 50 mL pure water at 40° C. was immediately introduced to this flask, and a homogeneous solution with obvious blue opalescence was formed upon shaking. The solution was filtered over 0.22 μm filtration membrance, and freeze-dried into a white powder in the lyophilizer. The content of dexamethasone in the solid was 30% as detected by HPLC.

(144) The lyophilized powder was redissolved by physiological saline to a solution with the concentration of 2 mg/mL. The solution was stable for more than 72 h at room temperature.

Example 61: Preparation of the Lyophilized Polymeric Micellar Formulation of Olaparib

(145) 200 mg amphiphilic block copolymer of formula Iz and 100 mg olaparib were added into 250 mL flask, and dissolved by 50 mL acetonitrile. The solution was placed in the shaker and shaked for 2 h at room temperature. A transparent thin film was formed around the wall of the flask after evaporation of acetonitrile. 50 mL pure water at 40° C. was immediately introduced to this flask, and a homogeneous solution with obvious blue opalescence was formed upon shaking. The solution was filtered over 0.22 μm filtration membrance, and freeze-dried into a white powder in the lyophilizer. The content of olaparib in the solid was 33% as detected by HPLC.

(146) The lyophilized powder was redissolved by physiological saline to a solution with the concentration of 2 mg/mL. The solution was stable for more than 72 h at room temperature.

Example 62: Preparation of the Lyophilized Polymeric Micellar Formulation of Combretastatin

(147) 200 mg amphiphilic block copolymer of formula Iaa and 70 mg combretastatin were added into 250 mL flask, and dissolved by 50 mL dichloromethane. The solution was placed in the shaker and shaked for 2 h at room temperature. A transparent thin film was formed around the wall of the flask after evaporation of dichloromethane. 50 mL pure water at 40° C. was immediately introduced to this flask, and a homogeneous solution with obvious blue opalescence was formed upon shaking. The solution was filtered over 0.22 μm filtration membrance, and freeze-dried into a white powder in the lyophilizer. The content of combretastatin in the solid was 25% as detected by HPLC.

(148) The lyophilized powder was redissolved by physiological saline to a solution with the concentration of 2 mg/mL. The solution was stable for more than 72 h at room temperature.

Example 63 Pharmacokinetics in Rats

(149) 1.1 Animals

(150) Male SD rats (6-8 weeks, 200-250 g) were obtained from Shanghai Sippr-BK laboratory animal Co., Ltd., with animal certificate number 2008001682093.

(151) 1.2 Sample Preparation

(152) 1) Micellar paclitaxel: paclitaxel lyophilized powder prepared from Example 46 of the present invention. The weight ratio of paclitaxel to copolymer Ia (PEG-linker-PLA) is 30:100.

(153) 2) Genexol-PM: lyophilized powder of paclitaxel-load micelle of Samyang. The weight ratio of paclitaxel to copolymer (PEG-PLA) is 20:100.

(154) 3) Paclitaxel injection: purchased from Beijing SL Pharmaceutical Co., Ltd., 30 mg/5 mL*10, batch number: 20170501.

(155) Lyophilized powder of micellar paclitaxel or Genexol-PM: An appropriate amount of normal saline was added to an appropriate amount of sample. The mixture was shaken at a speed of 200 r/min in shaker for about 20 minutes until it becomes clear prior to intravenous injection.

(156) 1.3 Drug Administration

(157) Intravenous injection: 3 male SD rats were administered intravenously for each compound after overnight fasting, at a dose of 3 mg/kg, with a volume of 3 mL/kg.

(158) 1.4 Methods

(159) Pre-dose and 0.0833, 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours post-dose, blood samples of each animal were collected by jugular vein puncture (approximately 0.15 mL at each time point). All collected samples were transferred to a pre-chilled EDTA-K2 tube or a pre-chilled plastic microcentrifuge tube containing 3 μL 0.5M EDTA-K2 as an anticoagulant and were placed on wet ice until centrifuged. Each collected blood sample was centrifuged at 4° C. for 15 minutes, and the plasma was collected. All plasma was stored in a freezer at about −80° C. until LCMS/MS detection.

(160) 1.5 Pharmacokinetic Results

(161) The pharmacokinetic parameters of each group are set forth in the following Table 1.

(162) TABLE-US-00001 TABLE 1 Micellar paclitaxel Paclitaxel PK parameters (from example 46) Genexol-PM injection Dose i.v. (3 mg/kg) i.v. (3 mg/kg) i.v. (3 mg/kg) C.sub.0 (ng/mL)  415 ± 63.2 547 ± 191 3224 ± 140  T.sub.1/2 (h) 8.96 ± 4.08 5.84 ± 1.52 6.54 ± 0.59 AUC.sub.0-36h (ng .Math. h/mL) 424 ± 125 457 ± 163 2576 ± 54.8  AUC.sub.0-inf (ng .Math. h/mL) 465 ± 148 501 ± 142 2612 ± 48.5  Cl (mL/min/kg) 92.8 ± 25.1  105 ± 28.8 19.1 ± 0.35 Vdss (L/kg) 46.2 ± 9.10 39.6 ± 5.06 5.51 ± 0.57

(163) 1.6 Conclusion

(164) 1) A single intravenous injection of an equal dose (10 mg/kg) was given to SD rats. The plasma exposure to paclitaxel from the micellar paclitaxel group of the present disclosure is significantly lower than that of paclitaxel injection group and that of Genexol-PM (Samyang, Korea) group, which indicates that the stability of the micellar paclitaxel of the present disclosure in vivo is significantly superior to that of the Genexol-PM (Samyang, Korea) and paclitaxel injection, and that the micellar paclitaxel of the present disclosure may be safer.

(165) 2) The half-life of paclitaxel in the micellar paclitaxel group of the present disclosure is significantly higher than that of paclitaxel injection group and Genexol-PM (Samyang, Korea) group, indicating that the micellar paclitaxel of the present disclosure may possess better therapeutic effects.

Example 64 Pharmacodynamic Study of Micellar Paclitaxel of this Invention on a Mouse Colo-205 Model

(166) 2.1 Animals

(167) Female BALb/c nude mice (5 weeks, 14-16 g) were obtained from Shanghai Sippr-BK laboratory animal Co., Ltd., with animal certificate number 20130016001491.

(168) 2.2 Feeding Conditions

(169) Animals are fed in the experimental environment once arrived for 7 days before the experiment starting. Animals were housed in SPF animal house with IVC (Individually Ventilated Cages, 4 mice per cage) system. Animal information card of each cage indicated the number, sex, strain, date received, dosing schedule, experiment No., group, and experiment start date of animals in the cage. All of the cages, bedding and drinking water were sterilized before use. Cages, feed and water were changed twice a week.

(170) 2.3 Establishment of Xenograft Model

(171) Human colorectal cancer Colo-205 cells (ATCC-CCL-222) were cultured in monolayer in vitro, in RPMI 1640 medium with 10% fetal bovine serum, 100 U/mL penicillin and 100 μg/mL streptomycin, in a incubator at 37° C. and 5% CO2. Routine digestion with trypsin-EDTA was performed twice a week. When the cell density reached 80%-90%, the cells were collected and counted. 0.2 mL (5×10.sup.6 cells) Colo-205 cells were subcutaneously inoculated on the right back of each mouse. When the average tumor volume reached 163 mm.sup.3, animals were randomized into groups.

(172) 2.4 Sample Preparation

(173) 1) Micellar paclitaxel: paclitaxel lyophilized powder prepared from Example 46 of the present disclosure. The weight ratio of paclitaxel to copolymer Ia (PEG-linker-PLA) is 30:100.

(174) 2) Genexol-PM: lyophilized powder of paclitaxel-load micelle of Samyang. The weight ratio of paclitaxel to copolymer (PEG-PLA) is 20:100.

(175) 3) Paclitaxel injection: purchased from Beijing SL Pharmaceutical Co., Ltd., 30 mg/5 ml*10, batch number: 20170501.

(176) Lyophilized powder of micellar paclitaxel or Genexol-PM: An appropriate amount of normal saline was added to an appropriate amount of sample. The mixture was shaken at a speed of 200 r/min in shaker for about 20 minutes until it became clear prior to intravenous injection.

(177) 2.5 Drug Administration

(178) Dosing schedules are presented in Table 2. Tumor volume under the skin, and weight of nude mice was measured 2-3 times a week.

(179) TABLE-US-00002 TABLE 2 Dose Administration No. Group (mg/kg) Route Quality Cycle 1 Vehicle 15 IV 6 IV × 14 days 2 Micellar 15 IV 6 IV × 14 days paclitaxel 3 Genexol-PM 15 IV 6 IV × 14 days 3 Paclitaxel 15 IV 6 IV × 14 days injection

(180) Note: Dosing volume was calculated based on body weight. The dosing volume was 10 μL/g.

(181) 2.6 Analysis and Evaluation

(182) Evaluation index: The tumor growth inhibition TGI (%) ratio or relative tumor proliferation T/C (%) ratio were used for evaluation, where T represents the experimental group and C represents the control group.

(183) Calculation of T/C (%): If T>T.sub.0, T/C (%)=(T−T.sub.0)/(C−C.sub.0)×100%, if T<T.sub.0, T/C (%)=(T−T.sub.0)/T.sub.0×100%, where T and C represent tumor volumes at the end of the experiment, T.sub.0 and C.sub.0 represent tumor volumes at the beginning of the experiment.
Calculation of rate TGI (%): TGI (%)=(1−T/C)×100%.

(184) Evaluation criteria: Activity was defined as T/C (%)≤40 (that is, TGI (%)≥60%), and P<0.05 of statistical test.

(185) 2.7 Pharmacological Results

(186) The inhibitory effects of micellar paclitaxel, Genexol-PM and paclitaxel injection on the Colo-205 tumor volume are shown in Table 3 and FIG. 17, and the weight change of nude mice is shown in FIG. 18.

(187) TABLE-US-00003 TABLE 3 Inhibition effect of each group on Colo-205 tumor volume Tumor Tumor volume volume (mm.sup.3) (mm.sup.3) T/C TGI P Group route (day 0) (day 14) (%) (%) value Vehicle iv 198 ± 12 1292 ± 129 — — — Micellar iv 198 ± 10  16 ± 5 −91.92 191.92 0.024 paclitaxel (15 mg/kg) Genexol-PM iv 198 ± 13  72 ± 14 −63.64 163.64 0.06 (15 mg/kg) Paclitaxel iv 198 ± 15  103 ± 21 −47.98 147.98 0.01 injection (15 mg/kg)

(188) The results show:

(189) 1) The micellar paclitaxel of the present disclosure, Genexol-PM, and paclitaxel injection notably inhibits tumor growth of Colo-205 xenografted tumors. The efficacy of the micellar paclitaxel group of the present disclosure is better than that of the Genexol-PM group and paclitaxel injection group

(190) 2) In paclitaxel injection groups, the animals had dysuria and some death happened. Rupture of bladder were found after dead animals were dissected. All the animals in the micellar paclitaxel group of the present disclosure appeared normal, indicating that the safety of the micellar paclitaxel of the present invention is better than that of paclitaxel injection.

Example 65 Pharmacodynamic Study of Micellar Paclitaxel of this Invention on a Mouse MCF-7 Model

(191) 3.1 Animals

(192) Female BALb/c nude mice (6-8 weeks, 18-20 g) were obtained from Shanghai Lingchang Biotech Co., Ltd., with animal certificate number 2013001829943.

(193) 3.2 Feeding Conditions

(194) Same as Example 2.2.

(195) 3.3 Establishment of Xenograft Model

(196) Human breast cancer MCF-7 cells (ECACC 86012803) were cultured in monolayer in vitro, in EMEM(EBSS) medium with 2 mM Glutamine, 1% Nonessential Amino Acids (NEAA), 10% fetal bovine serum, 100 U/mL penicillin and 100 μg/mL streptomycin, in a incubator at 37° C. and 5% CO.sub.2. Routine digestion with trypsin-EDTA was performed twice a week. When the cell density reached 80%-90%, the cells were collected and counted. 0.2 mL (1×10.sup.7 cells) MCF-7 cells mixing with Matrigel(v:v=1:1) were subcutaneously inoculated on the right back of each mouse. When the average tumor volume reached 209 mm.sup.3, animals were randomized into groups.

(197) 3.4 Sample Preparation

(198) 1) Micellar paclitaxel: paclitaxel lyophilized powder prepared from Example 36 of the present invention. The weight ratio of paclitaxel to copolymer Ia (PEG-linker-PLA) is 30:100.

(199) 2) Genexol-PM: lyophilized powder of paclitaxel-load micelle of Samyang. The weight ratio of paclitaxel to copolymer (PEG-PLA) is 20:100.

(200) 3) Paclitaxel injection: purchased from Beijing SL Pharmaceutical Co., Ltd., 30 mg/5 mL*10, batch number: 20170501.

(201) Lyophilized powder of micellar paclitaxel or Genexol-PM: An appropriate amount of normal saline was added to an appropriate amount of sample. The mixture was shaken at a speed of 200 r/min in shaker for about 20 minutes until it became clear prior to intravenous injection.

(202) 3.5 Drug Administration

(203) Dosing schedules are presented in Table 3. Tumor volume under the skin, and weight of nude mice was measured 2-3 times a week.

(204) TABLE-US-00004 TABLE 4 Dose Administration No. Group (mg/kg) route Quality Cycle 1 Vehicle 15 IV 6 IV × 14 days 2 Micellar 15 IV 6 IV × 14 days paclitaxel 3 Genexol-PM 15 IV 6 IV × 14 days 3 Paclitaxel 15 IV 6 IV × 14 days injection

(205) Note: Dosing volume was calculated based on body weight. The dosing volume was 10 μL/g.

(206) 3.6 Analysis and Evaluation

(207) Evaluation index: The tumor growth inhibition TGI (%) ratio or relative tumor proliferation T/C (%) ratio was used for evaluation, where T represents the experimental group and C represents the control group.

(208) Calculation of T/C (%): If T>T.sub.0, T/C (%)=(T−T.sub.0)/(C−C.sub.0)×100%, if T<T.sub.0, T/C (%)=(T−T.sub.0)/T.sub.0×100%, where T and C represent tumor volumes at the end of the experiment, T.sub.0 and C.sub.0 represent tumor volumes at the beginning of the experiment.
Calculation of rate TGI (%): TGI (%)=(1−T/C)×100%.

(209) Evaluation criteria: Activity was defined as T/C (%)≤40 (that is, TGI (%)≥60%), and P<0.05 of statistical test.

(210) 3.7 Pharmacological Results

(211) The inhibitory effects of micellar paclitaxel of the present disclosure, Genexol-PM and paclitaxel injection on the MCF-7 tumor volume are set forth in Table 4 and FIG. 19, and the weight change of nude mice is set forth in FIG. 20.

(212) TABLE-US-00005 TABLE 5 Inhibition effect of each group on MCF-7 tumor volume Tumor Tumor volume volume (mm.sup.3) (mm.sup.3) T/C TGI P Group route (day 0) (day 14) (%) (%) value Vehicle iv 191 ± 15 353 ± 5 — — — Micellar paclitaxel iv 191 ± 18  45 ± 4 −76.44 176.44 0.018 (15 mg/kg) Genexol-PM iv 191 ± 15  60 ± 9 −68.59 168.59 0.015 (15 mg/kg) Paclitaxel injection iv 191 ± 17  65 ± 5 −65.97 165.97 0.01 (15 mg/kg)

(213) The results showed:

(214) 1) The micellar paclitaxel of the present disclosure, Genexol-PM, and paclitaxel injection significantly inhibits tumor growth of MCF-7 xenografted tumors. The efficacy of the micellar paclitaxel group of the present disclosure is better than that of the other two groups.

(215) 2) In paclitaxel injection groups, the animals had dysuria and some death happened. Rupture of bladder were found after dead animals were dissected. All the animals in the micellar paclitaxel group of the present disclosure appeared normal, indicating that the safety of the micellar paclitaxel of the present disclosure is better than that of paclitaxel injection.

(216) List of abbreviations in the present disclosure is shown in table 6

(217) TABLE-US-00006 TABLE 6 Abbreviation Full name DBU 1,8-Diazabicyclo[5.4.0]undec-7-ene Me-PEG Polyethyleneglycol monomethyl ether PLA Polylactic acid/Polylactide Boc t-Butoxycarbonyl Fmoc 9-Fluorenylmethyloxycarbonyl TBSCl Tertbutyldimethylsilyl chloride DCC Dicyclohexylcarbodiimide NHS N-hydroxy succinimide EDCI 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide Hydrochloride DMAP N,N-dimethylaminopyridine Mn Number-average molecular weight

(218) Structural formulas of some compounds in the Examples are shown in table 7

(219) TABLE-US-00007 TABLE 7 No. Formula Ia Ib Ic 0 Id Ie If Ig Ih Ii Ij Ik Il Im 0 In Io Ip Iq Ir Is It Iu Iv Iw 0 Ix Iy Iz Ia′ Iaa IIa IIb IIc IId IIe 0 IIIa IIIb IIIc IIIe IVa IVb Va Vb VIa Va′ 00 IVa′ 01

(220) In the above table, the carbon with star (“*”) is chiral carbon. The configuration of the chiral carbon can be R, S or racemic.

(221) While the present disclosure has been described with reference to the specific examples, some modifications and equivalent variations will be apparent to those skilled in the art and are also within the scope of the present disclosure.