# Patent

## Title
Water-soluble taxane derivatives and uses thereof

## Abstract

The present invention discloses a class of water soluble taxane derivatives, a method for treating tumor using the same, use thereof as anti-tumor drugs and use thereof in the preparation of anti-tumor drugs. The water soluble taxane derivatives have the general formula (I-1), (I-2) or (I-3): wherein, R1 is H or methyl; R2 is H, methyl or acetyl; R3 is phenyl or OC(CH3)3; X is H, C1-6 alkyl or F; Y is F or C1-6 alkyl substituted with one or more F; n is 1, 2, 3, 4, 5 or 6; W is NR4R5.A or




R4 and R5 are each independently H, C1-6 alkyl optionally substituted with phenyl, or C3-6 cycloalkyl; m is 0, 1, 2 or 3; and A is a pharmaceutically acceptable acid.

## Background
CROSS-REFERENCE TO RELATED APPLICATIONS
The present application is a U.S. National Stage Filing under 35 U.S.C. 371 from International Application No. PCT/CN2015/073178, filed on Feb. 16, 2015, and published as WO 2015/120822 on Aug. 20, 2015, which claims the benefit of priority from Chinese patent application No. 201410053129.9 filed on Feb. 17, 2014, Chinese patent application No. 201410102551.9 filed on Mar. 19, 2014, and Chinese patent application Nos. 201410154956.7 and 201410155204.2 filed on Apr. 17, 2014, the contents of which are incorporated herein by reference.
TECHNICAL FIELD
The present invention relates to the field of pharmacy, specifically to water soluble taxane derivatives, a method for treating tumor using the same, use thereof as antitumor drugs, and use thereof in the preparation of antitumor drugs.
BACKGROUND ART
A prodrug, also referred to as a precursor of a drug, refers to a compound which achieves pharmacological action after the conversion in an organism. A prodrug per se has no or little bioactivity, and releases an active agent after metabolism in vivo. The purpose of investigating and preparing a prodrug is to increase the bioavailability, modify the solubility, enhance the targeting properties, or reduce the toxicity or side effects of the parent drug. It is advantageous for many drugs, especially those having low bioavailability, poor water solubility or high toxic side effects, to be prepared into prodrugs. In general, it is required in clinic that a prodrug can be quickly dissociated into a ligand and a parent drug after entering the body, and the ligand is non-toxic. The parent drug thus released can exert pharmaceutical effects, and the non-toxic ligand is of no harm to the body.
Taxane drugs (including those in clinical stage) with antitumor activity have the following parent structures:




wherein R1 is H or methyl; R2 is H, methyl or acetyl; R3 is phenyl or OC(CH3)3.

Among these drugs, paclitaxel, docetaxel and cabazitaxel are most widely used in clinic.

















Name
Parent Structure
R1
R2
R3









paclitaxel
(II-1)
H
Ac
Ph



docetaxel
(II-1)
H
H
OC(CH3)3



cabazitaxel
(II-1)
Me
Me
OC(CH3)3



larotaxel
(II-2)

Ac
OC(CH3)3



tesetaxel
(II-3)


OC(CH3)3










Taxane drugs have good antitumor activity. However, they have poor water solubility and low oral bioavailability, and thus can only be administered via intravenous injection in clinic. In clinic, nonionic surfactants, such as polyethoxylated castor oil or Tween-80, are often added to this class of compounds as co-solvents to prepare an injection. Unfortunately, such surfactants usually cause side effects, such as vasodilatation, reduced blood pressure, liver toxicity, severe allergic reactions and the like, which highly limit the clinical application of taxane drugs. As such, many pharmacists worldwide have tried to solve this problem by formulation improvement and structural modification.
With respect to the preparations of paclitaxel, references can be referred to the following: in 1995, Robert et al. described “Stable oil-in-water emulsions incorporating a taxane (taxol) and method of making same” (CN 1153474); in 1996, Hairu Zhang et al. described “Water soluble powder for injection of paclitaxel and preparation method thereof” (CN 96112502); in 1998, J. M. Géczy described “Pharmaceutical compositions containing cyclodextrins and taxoids” (Application No./Patent No. 98811010); in 1999, Rubinfeld, Joseph et al. described “Water-miscible pharmaceutical compositions of paclitaxel” (CN99812662), Guoying Weng et al. described “Lipid compositions of paclitaxel and preparation method thereof” (CN0019039), Yingjin Yuan et al. described “Prodrugs of paclitaxel or docetaxel supported by polyethylene glycol” (CN00109748), in 2001, Junqi Pan et al. described “A nanosized magnetic targeting preparation of paclitaxel and preparation method thereof” (CN01128733); in 2002, Bin Zhu et al. described “A preparation method of a nanoparticle of paclitaxel” (CN02133333), Jialin Yan et al. described “A novel preparation of an antitumor drug—a microemulsion of paclitaxel” (CN02153674); in 2003, Yu Liu et al. described “A water soluble complex of an antitumor drug, paclitaxel, and preparation method thereof” (CN03119497); in 2006, Xianghua Liu et al. described “A composition of paclitaxel/docetaxel liposomes and preparation method thereof” (CN200610137900), Shihai Li et al. described “A lipid microsphere injection of paclitaxel and preparation method thereof” (CN200610165800), Xiuguo Zhang et al. described “An intravenous injection solution of paclitaxel and application thereof” (CN200610165508); in 2007, Yunqing Kang et al. described “A paclitaxel loaded sustained release microsphere prepared by supercritical fluids technology” (CN200710049845), Fang Li et al. described “A nanoparticle of paclitaxel and preparation and application thereof” (CN200710047767); and in 2008, Dingquan Yao et al. described “A water soluble injection composition of paclitaxel, preparation and application thereof” (CN200810232882), Yuling Liu et al. described “A lipid composition of paclitaxel” (CN200810168213). However, all the above preparations have several disadvantages, and the clinical application thereof is limited.
With respect to the preparations of docetaxel, references can be referred to the following: Immordino et al. prepared liposomes containing docetaxel through a film dispersion method (Immordino M L, Brusa P, Arpicco S. et al., Preparation, characterization, cytotoxicity and pharmacokinetics of liposomes containing docetaxel, J. Controlled Release, 2003, 91 (4):417-429); LIVERSIDGE, Gary et al. prepared nanoparticulate preparations of docetaxel or analogue thereof (CN200680012670.9); Shaohui Zheng et al. investigated sub-microemulsion for intravenous injection of docetaxel and preparation method thereof (CN200610012102); Yuqing Xu et al. prepared a magnetic microsphere of docetaxel by a heat-curing method (Yuqing Xu, Xuemei Wen, A preparation method of a magnetic microsphere of docetaxel, Journal of Harbin Medical University, 2005, 39(6):537-539; Yuqing Xu, Xuemei Wen, a magnetic microsphere of docetaxel and preparation method thereof, CN200410044113); Zhenxin Du et al. prepared a lipid emulsion of docetaxel (Zhenxin Du, Xiulian Lu, Datao Li et al., A lipid emulsion containing docetaxel and preparation method thereof, CN 200510084055); Jakateet al. prepared fibrlnogen-coated olive oil droplets with a drug loading of 1.0 g/l (Jakate A S, Einhaus C M, DeAnglis A P. et al., preparation, characterization, and preliminary application of fibrinogen-coated olive oil droplets for the targeted delivery of docetaxel to solid malignancies, Cancer Res, 2003, 63(21):7314-7320); and Le Garreeet al. prepared a PVP-b-PDLLA polymeric micelle of docetaxel (Le Garree D, Gori S, Luo L. et al., Poly(N-vinylpyrrolidone)-block-poly(d,l-lactide) as new polymeric solubilizer for hydrophobic anticancer drugs: in vitro and in vivo evaluation, J Controlled Release, 2004, 99(1):83-101). All the above technologies are attempts for solving problems of taxane drugs currently used in clinic by formulation means, but with limited success.
The only successful water soluble preparation of taxane drugs is Albumin-bound paclitaxel marketed in 2005. Although this preparation does not contain a high molecular solvent, bubbles can be easily produced during formulating before injection due to the surfactant properties of albumin, and thus its application is not convenient.
Meanwhile, pharmaceutical chemists have focused on structural modification of taxane drugs. The main strategy for preparing water soluble derivatives of taxane drugs is to introduce hydrophilic groups to the 2′-OH or 7-OH in paclitaxel. Due to the high steric hindrance of 7-OH in paclitaxel, dissociation is difficult to occur after introduction of the groups at this position, and thus the compound obtained is unlikely to be used as a prodrug. The 2′-OH in taxane drugs is considered as an essential pharmacological group, and introduction of the groups to this position would result in decrease of efficacy. But the steric hindrance at this position is low, which would facilitate the dissociation. Pharmaceutical chemists have prepared many derivatives of paclitaxel and docetaxel (see e.g., Margri N F, Kingston D G I. J Nat Prod, 1988, 51:298; Journal of Medicinal Chemistry (1989), 32(4), 788-92; U.S. Pat. No. 4,960,790; Zhao Z. Kingston D G I. J Nat Prod, 1991, 54:1607; Mathew A, Mejillano M R, et al., J Med Chem, 1992, 35:4230; Nicolaon K C, Riemer C, Kerr M A, et al., Nature, 1993, 364:464; Nicolaon K C, Guy R K, Pitsinos E N, et al., Angew Chem Int Ed Engl, 1994, 33:1583; Chemistry & Biology 1995, 2 (4):223-227; JP09110865; J. Med Chem. 2000, 43, 3093-3102; Mendleev. Commun., 2001, 11(6):276-217; Biological & Pharmaceutical Bulletin (2002), 25(5), 632-641; J. Org. Chem., 2003, 68, 4894-4896; U.S. Pat. No. 6,649,778; CN200410002722; JP2006193627; Alaoui A. E., Saha N., Schmidt F., Monneret C. New Taxol (paclitaxel) prodrugs designed for ADEPT and PMT strategies in cancer chemotherapy, Bioog Med. Chem., 2006, 14:5012-5019; Bioorg. Med. Chem. Lett. 17 (2007) 2894-2898; Pharmaceutical Research, 26(4):785-793, 2009; Bioconjugate Chem. 2009, 20, 2214-2221; Pharmaceutical Research, 2010, 27(2):380-389; J. Med. Chem., 1997, 40(26):4319-28; J. Med Chem., 1996, 39(7):1555-9; Journal of Tianjin University, 2000, 33(1):51-5; Chemical Journal of Chinese Universities, 2000, 21(3): 401-6; EP524093, 1993; WO9623779; WO9802426; U.S. Pat. No. 6,025,385; WO9914209; U.S. Pat. No. 6,136,808; US20020052403; CN200610171580; Wenting Du, Lan Hong, Tongwei Yao, et al, Synthesis and evaluation of water-soluble docetaxelprodrugs-docetaxel esters of malic acid, Bioorganic & Medicinal Chemistry, 2007, 15(18), 6323-6330; CN200510027736.9; CN200510040320; WO2009141738; Chemistry & Biology 1995, 2 (4):223-227). However, no further reports regarding the derivatives have been found.
Since no substantial progress has been achieved in the investigations of water soluble derivatives of paclitaxel and docetaxel, no reports on the water soluble derivatives of subsequent taxane drugs, i.e., cabazitaxel, larotaxel and tesetaxel, have been found.
Although much effort has been made to the structural modification of taxane drugs by pharmaceutical chemists, no taxane derivative having good water solubility and easy dissociation in vivo has been found.
SUMMARY OF THE INVENTION
In a first aspect, the present invention provides a carboxylic acid derivative or a salt thereof useful as a ligand for the preparation of a water soluble paclitaxel prodrug which has good water solubility and can be quickly dissociated in vivo and release the parent drug, so as to exert effects.
The carboxylic acid derivative of the present invention has following general formula (III):




wherein,

X is H, C1-6 alkyl or F;
Y is F or C1-6 alkyl substituted with one or more F;
n is 1, 2, 3, 4, 5 or 6;
W1 is NR4R4, NR4R5.B,




R4 and R5 are each independently H, C1-6 alkyl optionally substituted with phenyl, or C3-6 cycloalkyl;
m is 0, 1, 2 or 3;
B is an acid; and
D is hydroxyl, Cl or Br.
According to an embodiment of the present invention, X is H, methyl or F.
According to an embodiment of the present invention, Y is F, CF3 or CHF2.
According to an embodiment of the present invention, R4, R5 are each independently H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, benzyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.
According to an embodiment of the present invention, the acid B is an acid which can form a salt with amine, such as hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, trifluoroacetic acid, difluoroacetic acid, fluoroacetic acid, acetic acid, benzensulfonic acid or p-toluene sulfonic acid.
According to an embodiment of the present invention, when X and Y are different (i.e., the α-C of the carboxyl in the carboxylic acid derivative is a chiral atom), the carbon atom to which both X and Y are attached is in a single R configuration, in a single S configuration, or in both R and S configurations.
According to an embodiment of the present invention, the carboxylic acid derivative of the present invention is selected from the group consisting of:

    4-N,N-dimethylamino-2(R)-fluorobutyric acid hydrochloride;
    4-N-isopropylamino-2(R,S)-fluorobutyric acid hydrochloride;
    4-N,N-diethylamino-2(R,S)-trifluoromethylbutyric acid hydrochloride;
    4-N-benzylamino-2,2-difluorobutyric acid hydrochloride;
    4-N-isobutylamino-2(R,S)-difluoromethylbutyric acid hydrochloride;
    4-N-(aziridin-1-yl)-2(R,S)-difluoromethylbutyric acid hydrochloride;
    4-N-(pyrrolidin-1-yl)-2(R,S)-fluorobutyric acid hydrochloride;
    3-N-benzylamino-2(R,S)-(1,1-difluoromethyl)propionic acid hydrochloride;
    6-N-cyclohexylamino-2(R,S)-trifluoromethylhexanoic acid hydrochloride;
    sodium 4-N,N-dimethylamino-2(R,S)-fluorobutyrate;
    calcium 4-N, N-diethylamino-2(R,S)-fluorobutyrate;
    aluminum 3-N-benzylamnino-2(R,S)-benzyloxypropionate;
    4-N,N-dimethylamino-2(R,S)-fluorobutyryl chloride hydrochloride;
    4-N-benzylamino-2,2-difluorobutyryl chloride hydrochloride;
    4-N,N-dimethylamino-2(R,S)-fluorobutyric acid; and
    4-N,N-dimethylamino-2(S)-fluorobutyric acid hydrochloride.


In a second aspect, the present invention provides water soluble taxane derivatives. The water soluble taxane derivatives have stable chemical properties and good water solubility. These water soluble taxane derivatives are a series of amine-containing salt compounds prepared by a reaction between the hydroxyl group at position 2′ in taxane compounds and amino acids.
The water soluble taxane derivative of the present invention has following general formula (I-1), (I-2) or (I-3):




wherein,

R1 is H or methyl:
R2 is H, methyl or acetyl;
R3 is phenyl or OC(CH3)3;
X, Y and n are as defined above for the compound of general formula (III);
W is NR4R5.A or




R4, R5 and m are as defined above for the compound of general formula (III); and
A is a pharmaceutically acceptable acid.
According to an embodiment of the present invention, in general formula (I-1),
R1 is H, R2 is acetyl, and R3 is phenyl,
R1 is H, R2 is H, and R3 is OC(CH3)3; or
R1 is methyl, R2 is methyl, and R3 is OC(CH3)3.
According to an embodiment of the present invention, in general formula (I-2),
R2 is acetyl and R3 is OC(CH3)3.
According to an embodiment of the present invention, in general formula (I-3),
R3 is OC(CH3)3.
According to an embodiment of the present invention, X is H, methyl or F.
According to an embodiment of the present invention, Y is F, CF3 or CHF2.
According to an embodiment of the present invention, R4, R5 are each independently H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, benzyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.
According to an embodiment of the present invention, X is H, methyl or F; Y is F, CF3 or CHF2; R4, R5 are each independently H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, benzyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.
According to an embodiment of the present invention, A is hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, carbonic acid, acetic acid, propionic acid, methanesulfonic acid, lactic acid, benzensulfonic acid, p-toluene sulfonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid or malic acid.
According to an embodiment of the present invention, when X and Y are different (i.e., the α-C atom of the 2′-ester carbonyl in the water soluble taxane derivative is a chiral atom), the carbon atom to which both X and Y are attached is in a single R configuration, in a single S configuration, or in both R and S configurations.
According to an embodiment of the present invention, the water soluble taxane derivative is selected from the group consisting of:

    2′-O-[4-N,N-dimethylamino-2(R)-fluorobutyryl]paclitaxel hydrochloride;
    2′-O-[4-N,N-dimethylamino-2(R)-fluorobutyryl]paclitaxel methanesulfonate;
    2′-O-[4-N,N-dimethylamino-2(R,S)-fluorobutyryl]paclitaxel hydrochloride;
    2′-O-[4-N,N-dimethylamino-2(R,S)-difluoromethylbutyryl]paclitaxel citrate;
    2′-O-[4-N,N-dimethylamino-2(R)-fluorobutyryl]docetaxel hydrochloride;
    2′-O-[4-N,N-dimethylamino-2(R)-fluorobutyryl]docetaxel methanesulfonate;
    2′-O-[4-amino-2(R,S)-trifluoromethylbutyryl]docetaxel methanesulfonate;
    2′-O-[4-N,N-dimethylamino-2(R,S)-fluorobutyryl]cabazitaxel hydrochloride;
    2′-O-[4-N,N-dimethylamino-2(R)-fluorobutyryl]larotaxel hydrochloride;
    2′-O-[4-N,N-dimethylamino-2(R)-fluorobutyryl]tesetaxel hydrochloride;
    2′-O-[4-N,N-dimethylamino-2(S)-fluorobutyryl]paclitaxel hydrochloride;
    2′-O-[4-N,N-dimethylamino-2(S)-fluorobutyryl]docetaxel hydrochloride;
    2′-O-[4-N,N-dimethylamino-2(R,S)-fluorobutyryl]docetaxel hydrochloride;
    2′-O-[4-N,N-dimethylamino-2(R)-fluorobutyryl]cabazitaxel hydrochloride;
    2′-O-[4-N,N-dimethylamino-2(S)-fluorobutyryl]cabazitaxel hydrochloride;
    2′-O-[4-amino-2(R)-difluoromethylbutyryl]cabazitaxel sodium bisulfate salt;
    2′-O-[4-N,N-diethylamino-2-methyl-2(R)-2-trifluoromethylbutyryl]cabazitaxel methanesulfonate;
    2′-O-[4-N-methyl-N-ethylamino-2(R)-2-difluoroethylvaleryl]docetaxel hydrochloride;
    2′-O-[4-N,N-dimethylamino-2(R)-fluorovaleryl]docetaxel methanesulfonate;
    2′-O-[4-(aziridin-1-yl)amino-2,2-difluoroheptanoyl]docetaxel methanesulfonate;
    2′-O-[4-N-methyl-N-cyclopentylamino-2-trifluoromethyl-2-fluorooctanoyl]paclitaxel methanesulfonate;
    2′-O-[4-N,N-dimethylamino-2(R)-fluorovaleryl]larotaxel methanesulfonate;
    2′-O-[4-N,N-dimethylamino-2(R)-fluorovaleryl]tesetaxel methanesulfonate;
    2′-O-[4-N,N-dimethylamino-2(S)-fluorobutyryl]paclitaxel methanesulfonate;
    2′-O-[4-N,N-dimethylamino-2(R,S)-fluorobutyryl]paclitaxel methanesulfonate;
    2′-O-[4-N,N-dimethylamino-2(S)-fluorobutyryl]docetaxel methanesulfonate;
    2′-O-[4-N,N-dimethylamino-2(R,S)-fluorobutyryl]docetaxel methanesulfonate;
    2′-O-[4-N,N-dimethylamino-2(R)-fluorobutyryl]cabazitaxel methanesulfonate;
    2′-O-[4-N,N-dimethylamino-2(S)-fluorobutyryl]cabazitaxel methanesulfonate;
    2′-O-[4-N,N-dimethylamino-2(R,S)-fluorobutyryl]cabazitaxel methanesulfonate;
    2′-O-[4-amino-2(R)-difluoromethylbutyryl]cabazitaxel sulfate;
    2′-O-[4-N,N-diethylamino-2-methyl-2(R)-2-trifluoromethylbutyryl]cabazitaxel hydrochloride;
    2′-O-[4-N-methyl-N-ethylamino-2(R)-2-difluoroethylvaleryl]docetaxel methanesulfonate;
    2′-O-[4-N,N-dimethylamino-2(R)-fluorovaleryl]docetaxel hydrochloride;
    2′-O-[4-(aziridin-1-yl)amino-2,2-difluoroheptanoyl]docetaxel hydrochloride;
    2′-O-[4-N-methyl-N-cyclopentylamino-2-trifluoromethyl-2-fluorooctanoyl]paclitaxel hydrochloride;
    2′-O-[4-N,N-dimethylamino-2(R)-fluorovaleryl]larotaxel hydrochloride;
    2′-O-[4-N,N-dimethylamino-2(R)-fluorovaleryl]tesetaxel hydrochloride;
    2′-O-[4-N-methyl-N-ethylamino-2(R)-fluorobutyryl]paclitaxel methanesulfonate;
    2′-O-[4-N,N-diethylamino-2(S)-fluorobutyryl]paclitaxel fumarate;
    2′-O-[4-N-methyl-N-isopropylamino-2(R,S)-fluorobutyryl]paclitaxel hydrochloride;
    2′-O-[4-N,N-dimethylamino-2(R,S)-2-trifluoromethylbutyryl]paclitaxel p-toluene sulfonate;
    2′-O-[4-N,N-dimethylamino-2(R,S)-2-difluoromethylbutyrvl]paclitaxel hydrochloride;
    2′-O-[4-N,N-dimethylamino-2(R)-fluorovaleryl]paclitaxel maleate;
    2′-O-[4-N,N-dimethylamino-2(S)-2-difluoromethylhexanoyl]paclitaxel sulfate;
    2′-O-[4-N-methyl-N-ethylamino-2(R)-fluorobutyryl]docetaxel methanesulfonate;
    2′-O-[4-N,N-diethylamino-2(S)-fluorobutyryl]docetaxel maleate;
    2′-O-[4-N-methyl-N-isopropylamino-2(R)-fluorobutyryl]docetaxel hydrochloride;
    2′-O-[4-N,N-dimethylamino-2(R)-2-trifluoromethylbutyryl]docetaxel methanesulfonate;
    2′-O-[4-N,N-dimethylamino-2(R,S)-2-difluoromethylbutyrvl]docetaxel hydrochloride;
    2′-O-[4-N,N-dimethylamino-2(R,S)-fluorovaleryl]docetaxel methanesulfonate;
    2′-O-[4-N,N-dimethylamino-2(S)-fluorohexanoyl]docetaxel sulfate;
    2′-O-[4-N-methyl-N-ethylamino-2(R)-fluorobutyryl]cabazitaxel maleate;
    2′-O-[4-N,N-diethylamino-2(R)-fluorobutyryl]cabazitaxel methanesulfonate;
    2′-O-[4-N-methyl-N-isopropylamino-2(R)-fluorobutyryl]cabazitaxel hydrochloride;
    2′-O-[4-N,N-dimethylamino-2(R)-2-trifluoromethylbutyryl]cabazitaxel methanesulfonate;
    2′-O-[4-N,N-dimethylamino-2(R)-2-difluoromethylbutyryl]cabazitaxel p-toluene sulfonate;
    2′-O-[4-N,N-dimethylamino-2(R)-fluorovaleryl]cabazitaxel hydrochloride;
    2′-O-[4-N,N-dimethylamino-2(R)-fluorohexanoyl]cabazitaxel sulfate;
    2′-O-[4-N-methyl-N-ethylamino-2(R,S)-fluorobutyryl]larotaxel hydrochloride;
    2′-O-[4-N-methyl-N-isopropylamino-2(R,S)-2-trifluoromethylbutyryl]larotaxel maleate;
    2′-O-[4-N,N-dimethylamino-2(S)-fluorovaleryl]larotaxel phosphate;
    2′-O-[4-N,N-diethylamino-2(S)-fluorobutyryl]tesetaxel hydrochloride;
    2′-O-[4-N-methyl-N-isopropylamino-2(R,S)-2-trifluoromethylbutyryl]tesetaxel succinate;
    2′-O-[4-(pyrrolidin-1-yl)amino-2(R,S)-fluorovaleryl]tesetaxel sulfate;
    2′-O-[4-N,N-diethylamino-2(R)-fluorobutyryl]paclitaxel hydrochloride;
    2′-O-[3-N,N-dimethylamino-2(R,S)-fluoropropionyl]paclitaxel hydrochloride;
    2′-O-[4-N-benzylamino-2(S)-fluorobutyryl]docetaxel hydrochloride;
    2′-O-[3-(N,N-diethyl)amino-2(R,S)-trifluoromethyl propionyl]docetaxel hydrochloride;
    2′-O-[4-(N-methyl)amino-2(R)-trifluoromethylbutyryl]paclitaxel hydrochloride;
    2′-O-[3-(N-isopropyl)amino-2(R,S)-difluoromethylpropionyl]cabazitaxel hydrochloride;
    2′-O-[5-(N,N-dimethyl)amino-2(R)-fluorovaleryl]paclitaxel hydrochloride;
    2′-O-[4-(N,N-dimethyl)amino-2(S)-trifluoromethylbutyryl]cabazitaxel hydrochloride;
    2′-O-[4-(N-isopropyl)amino-2(R,S)-trifluoromethylbutyryl]paclitaxel hydrochloride;
    2′-O-[5-N,N-dimethylamino-2(S)-trifluoromethylvaleryl]docetaxel hydrochloride;
    2′-O-[4-N-benzylamino-2-methyl-2(R,S)-fluorobutyryl]paclitaxel hydrochloride;
    2′-O-[4-N,N-dimethylamino-2(R)-difluoromethylbutyryl]docetaxel hydrochloride;
    2′-O-[3-cyclopentylamino-2-ethyl-2(R,S)-trifluoromethylpropionyl]docetaxel hydrochloride;
    2′-O-[5-N-benzylamino-2-benzyl-2(R)-difluoromethylvaleryl]cabazitaxel hydrochloride;
    2′-O-[4-(4-piperidin-1-yl)-2(S)-trifluoromethylbutyryl]cabazitaxel hydrochloride;
    2′-O-[4-N,N-dimethylamino-2(S)-fluorobutyryl]larotaxel hydrochloride;
    2′-O-[4-N,N-dimethylamino-2(R)-fluorobutyryl]tesetaxel hydrochloride.




DETAILED DESCRIPTIONS OF THE INVENTION
Definitions
The term “C1-6alkyl” as used herein refers to a saturated, linear or branched hydrocarbon group having 1-6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neo-pentyl, n-hexyl, isohexyl and the like, preferably methyl, ethyl, propyl, isopropyl, butyl or isobutyl, more preferably methyl, ethyl or propyl.
The term “C3-6 cycloalkyl” as used herein refers to a saturated monocyclic hydrocarbon group having 3-6 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.
The term “an acid which can form a salt with an amine” as used herein refers to an inorganic or organic acid commonly used in the field of organic chemistry which can form a salt with an amine. The inorganic acid includes, but is not limited to, hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, pyrosulfuric acid, phosphoric acid, nitric acid and the like. The organic acid includes, but is not limited to, formic acid, acetic acid, propionic acid, butyric acid, pivalic acid, trifluoroacetic acid, difluoroacetic acid, fluoroacetic acid, acetoacetic acid, benzoic acid, methanesulfonic acid, ethanesulfonic acid, trifluoromethanesulfonic acid, benzensulfonic acid, p-toluene sulfonic acid, naphthalenesulfonic acid, camphorsulfonic acid, and the like.
The term “pharmaceutically acceptable acid” as used herein refers to an acid which can be used in the medical field, such as hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, carbonic acid, acetic acid, propionic acid, methanesulfonic acid, lactic acid, benzensulfonic acid, p-toluene sulfonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid or malic acid.
Reaction Scheme
Compound (I-1) of the present invention can be prepared according to following Reaction Scheme 1:




Compound (I-2) of the present invention can be prepared according to following Reaction Scheme 2:




Compound (I-3) of the present invention can be prepared according to following Reaction Scheme 3:




wherein R1, R2, R3, X, Y, n, W, W1 and D are as defined above.
Step 1
The compound of general formula I-1a, I-2a or I-3a is prepared by reacting the compound of general formula II-1, II-2 or II-3 with the compound of general formula (III), respectively, in the presence of reagent (IV) at −100-40° C.
Step 2
The compound of general formula I-1 or I-2 is prepared by dissolving the corresponding compound of general formula I-1a or I-2a in an organic solvent, and then washing the organic layer with a saturated aqueous solution of a salt of acid A which is adjusted to below pH 5 with acid A; or
the compound of general formula I-3 is prepared by dissolving the corresponding compound of general formula I-3a in an organic solvent, and then washing the organic layer with a saturated aqueous solution of a salt of acid A which is adjusted to below pH 6 with acid A.
The organic solvent mentioned above refers to an aprotic organic solvent which can dissolve the compound of general formula I-1a, I-2a or I-3a, e.g., dichloromethane, chloroform, 1,2-dichloroethane, chlorobenzene, acetonitrile and the like.
The reagent (IV) mentioned above is a nitrogen-containing basic organic reagent or a mixed reagent consisting of a nitrogen-containing basic organic reagent and a compound having a carbodiimide structure. The nitrogen-containing basic organic reagent is an organic base, such as triethylamine, pyridine, DMAP (4-N,N-dimethylaminopyridine) or 4-PPY (4-pyrrolidinylpyridine); and the compound having a carbodiimide structure is DCC (dicyclohexylcarbodiimide) or EDC (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide).
When D in the compound of general formula (III) is Cl or Br, the reagent (IV) is a nitrogen-containing basic organic reagent, and the molar ratio of the compound of general formula (II) (i.e., the compound of II-1, II-2 or II-3), the compound of general formula (III) and reagent (IV). i.e., (II):(III):(IV), is 1:(1-12.0):(1-15.0).
When D in the compound of general formula (III) is OH, the reagent (IV) is a mixed reagent consisting of a nitrogen-containing basic organic reagent and a compound having a carbodiimide structure in a molar ratio of 1:(1-5), and the molar ratio of the compound of general formula (II) (i.e., the compound of II-1, II-2 or II-3), the compound of general formula (III) and the reagent (IV), i.e., (II):(III):(IV) is 1:(1-12.0):(1-15.0) (wherein the amount of the reagent (IV) is based on the amount of the nitrogen-containing basic organic reagent).
Specifically, the present invention provides a method for preparing a water soluble taxane derivative. The method comprises: dissolving a pharmaceutically active taxane compound (II-1, II-2 or II-3) in dichloromethane in the presence of pyridine or DMAP at −50° C.-50° C., slowly adding a solution of the compound of general formula (III) in dichloromethane dropwise, conducting separation after completion of the reaction, dissolving the product thus obtained in dichloromethane, and washing the organic layer with a saturated aqueous solution of a salt of acid A which is adjusted to an appropriate pH with acid A to obtain a water soluble taxane derivative of the present invention. The molar ratio of the pharmaceutically active taxane compound, the derivative of amino acid and pyridine or DMAP is 1:(1-10):(3-10.0).
The carboxylic acid derivative of general formula (III) of the present invention can be prepared according to following Reaction Scheme 4:




In the above Reaction Scheme 4, the compounds of formulae (IIIa), (IIIb) and (IIIc), which all belong to the compound of general formula (III), are obtained by reacting the compound of general formula (V) with an alkylating agent for amino (C1 or C3) or a protecting agent for amino (C2),
wherein,
R4, X, Y, n and B are as defined above;
C1 is an alkylating agent for amino, such as formic acid/formaldehyde, dimethyl sulfate, bromoethane, bromopropane, chlorobutane, acetone, butanone, cyclopentanone, cyclohexanone, benzaldehyde and the like;
C2 is a protecting agent for amino, such as benzyl chloroformate, di-tert-butyloxycarbonylcarbonic anhydride, benzyl chloride, benzyl bromide and the like;
C3 is another alkylating agent for amino, such as 1-chloro-2-bromoethane, 1-chloro-4-bromobutane, 1-chloro-5-bromopentane and the like;
R5a is alkyl or cycloalkyl, especially C1-6 alkyl, such as methyl, ethyl, propyl, isopropyl, butyl or isobutyl, or C3-6 cycloalkyl, such as cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.
R5b is an amino protecting group, especially C1-6 alkoxycarbonyl optionally substituted with phenyl (such as benzyloxycarbonyl or tert-butyloxycarbonyl) or benzyl.
When D in the compound of general formula (III) is Cl or Br, a corresponding acyl halide compound can be prepared from a starting material wherein D is hydroxyl via a halogenating reaction with a halogenating reagent (such as thionyl chloride, phosphorus trichloride, phosphorous pentachloride or phosphorus tribromide).
The compound of general formula (V) in the above reaction schemes can be obtained by methods reported in relevant references, e.g. [1] Chencomm, 1999: 1739-1740; [2] J. Med. Chem, 2011, 44:2849-2856; [3] JCS Perkin I 1980: 2029-2032; [4] Journal of Fluorine Chemistry (23), 1983: 241-259; [5] Journal of Fluorine Chemistry, 2004, vol. 125 (4): 509-515.
Use of the Water Soluble Taxane Derivative of the Present Invention in the Preparation of an Antitumor Drug
The water soluble taxane derivative of the present invention, as a prodrug, has the following beneficial effects:
The water soluble taxane derivative of general formula I-1, I-2 or I-3 of the present invention has higher water solubility than the corresponding compound of general formula II-1, II-2 or II-3, and thus can be formulated as a water soluble injection. After intravenous injection into an animal body, it can release the corresponding parent drug, so as to inhibit the growth of tumors.
The inventors prepare an ester derivative of a taxane drug through structural modification without changing the pharmacological activity of the parent drug, and form a salt thereof with a corresponding acid to facilitate dissociation in vivo. The water soluble taxane derivative of the present invention is relatively stable in chemical properties, and the aqueous solution thereof can release the parent drug after injection, thus overcoming the following disadvantages of currently available taxane preparations caused by the presence of polyethoxylated castor oil or Tween-80: acute hypersensitivity, neurotoxicity, liver toxicity, etc. In addition, the preparation method of the present invention is simple and safe to perform, and the quality of the product can be easily controlled.
EXAMPLES
To make the purpose and technical solutions of the present invention more clear, the preferable examples of the present invention are described in more detail as follows. It should be noted that the following examples are provided merely for further illustration of the present invention, but should not be construed to limit the scope of the present invention. Technical solutions obtained through non-essential improvements and adjustments by a person skilled in the art according to the above disclosure of the present invention all belong to the protection scope of the present invention, A. Preparation of the Carboxylic Acid Derivative of General Formula (III) of the Present Invention
Example 1
Preparation of 4-N,N-dimethylamino-2(R)-fluorobutyric acid hydrochloride
4-amino-2(R)-fluorobutyric acid hydrochloride (1.1 g, 7.0 mmol) was added to a round bottom flask, a saturated aqueous Na2CO3 solution was added to adjust the pH value to 8, and then 88% formic acid (6 ml) and 35% aqueous formaldehyde solution (5 ml) were added. The reaction mixture was warmed slowly to 80° C., and was allowed to react for 15 hours. The reaction mixture was cooled to room temperature, and 6 N hydrochloric acid (2 ml) was added followed by concentration under reduced pressure to give a light yellow solid. The solid was dissolved in methanol (10 ml), and the resulting solution was cooled in an ice bath with stirring for 30 min. Then the resulting mixture was filtered, and the filtrate was concentrated. The residue was refluxed in 6 N hydrochloric acid (100 ml) for 4 hours, and the liquid was removed by rotary evaporation. The solid thus obtained was treated with acetonitrile to obtain a white solid (1.1 g, yield: 85%).
m.p.: 136-138° C.:
1H-NMR (400 MHz, D2O): δ 4 72 (ddd, 1H), 2.90 (dtd, 2H), 2.43 (s, 6H), 1.93 (m, 2H);
13C-NMR (600 MHz, D2O): δ 173.13, 86.90, 53.49, 42.83, 26.91;
ESI-MS m/z [M+H]+ 150.13.
Example 2
Preparation of 4-N-isopropylamino-2(R,S)-fluorobutyric acid hydrochloride
4-amino-2(R,S)-fluorobutyric acid hydrochloride (1.1 g, 7.0 mmol) was added to a round bottom flask (50 ml), a saturated aqueous Na2CO1 solution was added to adjust the pH to 8, and then acetone (15 ml) and 5% Pd—C (100 mg) were added. Air was replaced with nitrogen, which was then replaced with hydrogen. The reaction was carried out for 6 hours at room temperature. Pd—C was removed through filtration, and the pH of the solution was adjusted to acidic with 6 N hydrochloric acid. The solution was concentrated under reduced pressure to give a light yellow solid. The solid was dissolved in methanol (10 ml), and the resulting solution was cooled in an ice bath with stirring for 30 min. Then the resulting mixture was filtered, and the filtrate was concentrated. The residue was refluxed in 6 N hydrochloric acid (100 ml) for 4 hours, and the solvent was removed by rotary evaporation. The solid thus obtained was treated with acetonitrile to obtain a white solid (1.05 g, yield: 75%).
ESI-MS m/z [M+H]+ 164.12.
Example 3
Preparation of 4-N,N-diethylamino-2(R,S)-trifluoromethylbutyric acid hydrochloride
4-amino-2(R,S)-trifluoromethylbutyric acid hydrochloride (2.07 g, 10 mmol) was added to a round bottom flask (50 ml), and 1 N aqueous NaHCO3 solution was added to adjust the pH value to 8. Acetonitrile (50 ml) was added, and a mixed solution (10 ml) of bromoethane (2.18 g, 20 mmol) and acetonitrile was added dropwise. The pH of the reaction solution was maintained at 7-8 with a solution of sodium bicarbonate. Hydrochloric acid was added to adjust the pH to below 5 after completion of the reaction, and the solution was concentrated under reduced pressure to give a light yellow solid. Methanol (10 ml) was added, the resulting solution was stirred for 30 min before filtration, and the filtrate was concentrated. The residue was refluxed in 6 N hydrochloric acid (100 ml) for 4 hours, the solvent was removed by rotary evaporation, and a white solid (yield: 130/0) was obtained.
ESI-MS m/z [M+H]+ 228.16.
Example 4
Preparation of 4-N-benzylamino-2,2-difluorobutyric acid hydrochloride
The title compound was prepared according to the method of Example 2, using 4-amino-2,2-difluorobutyric acid hydrochloride (1.1 g, 5.6 mmol) and benzaldehyde as starting materials.
ESI-MS nm/z [M+H]+ 230.06.
Example 5
Preparation of 4-N-isobutylamino-2(R,S)-difluoromethylbutyric acid hydrochloride
The title compound was prepared according to the method of Example 2, using 4-amino-2(R,S)-difluoromethylbutyric acid hydrochloride (1.90 g, 10 mmol) and butanone (15 ml) as starting materials, and a white solid (1.1 g, yield: 45%) was obtained.
m.p.: 141-142° C.;
ESI-MS m/z [M+H]+ 210.1.
Example 6
Preparation of 4-N-(aziridin-1-yl)-2(R,S)-difluoromethylbutyric acid hydrochloride
4-amino-2(R,S)-difluoromethylbutyric acid hydrochloride (1.90 g, 10 mmol) was added to a round bottom flask (50 ml), an aqueous NaHCO3 solution was added to adjust the pH to 7-8, and acetonitrile (15 ml) and 1-chloro-2-bromoethane (10 mmol) were added. The reaction was carried out at ambient temperature for 0.5 h, and then the reaction mixture was heated to reflux and was allowed to react under reflux for 2 h. The solvent was removed by evaporation under reduced pressure, and methanol (10 ml) was added to the residue. The resulting solution was then cooled in an ice bath with stirring for 30 min before filtration, and the filtrate was concentrated. The residue was refluxed in 6 N hydrochloric acid (100 ml) for 4 hours, the solvent was removed by rotary evaporation, and a white solid (0.7 g) was obtained.
ESI-MS m/z [M+H]+ 180.14.
Example 7
Preparation of 4-N-(pyrrolidin-1-yl)-2(R,S)-fluorobutyric acid hydrochloride
The title compound was prepared according to the method of Example 6, using 4-amino-2(R,S)-fluorobutyric acid and 1-chloro-4-bromobutane as starting materials.
ESI-MS m/z [M+H]+ 176.1.
Example 8
Preparation of 3-N-benzylamino-2(R,S)-(1,1-difluoromethyl)propionic acid hydrochloride
The title compound was prepared according to the method of Example 2, using 3-amino-2(R,S)-(1,1-difluoromethyl)propionic acid hydrochloride and benzaldehyde as starting materials.
ESI-MS m/z [M+H]+ 230.19.
Example 9
Preparation of 6-N-cyclohexylamino-2(R,S)-trifluoromethylhexanoic acid hydrochloride
The title compound was prepared according to the method of Example 2, using 6-amino-2(R,S)-trifluoromethylhexanoic acid hydrochloride and cyclohexanone as starting materials.
ESI-MS m/z [M+H]+ 282.08.
According to the above examples, the inventors also prepared the following compounds:

    sodium 4-N,N-dimethylamino-2(R,S)-fluorobutyrate (ESI-MS m/z [M−H] 149.03);
    calcium 4-N,N-diethylamino-2(R,S)-fluorobutyrate (ESI-MS m/z [M−H] 177.08);
    aluminum 3-N-benzylamino-2(R,S)-benzyloxypropionate (ESI-MS m/z [M−H] 197.04);
    4-N,N-dimethylamino-2(R,S)-fluorobutyryl chloride hydrochloride (ESI-MS m/z [M−H] 168.01);
    4-N-benzylamino-2,2-difluorobutyryl chloride hydrochloride (ESI-MS m/z [M−H] 248.03);
    4-N,N-dimethylamino-2(R,S)-fluorobutyric acid (ESI-MS m/z [M−H] 150.08); and
    4-N,N-dimethylamino-2(S)-fluorobutyric acid hydrochloride (ESI-MS m/z [M−H] 150.11).


B. Preparation of the Water Soluble Taxane Derivative of the Present Invention
B-1. Preparation of Paclitaxel Derivatives
Example 10
2′-O-[4-N,N-dimethylamino-2(R)-fluorobutyryl]paclitaxel hydrochloride (compound 01)
1) The preparation of 4-N,N-dimethylamino-2(R)-fluorobutyryl chloride hydrochloride: 4-N,N-dimethylamino-2(R)-fluorobutyric acid hydrochloride (10 mmol) was placed in thionyl chloride (10 ml) in a flask, the mixture was slowly warmed to 40° C., and the reaction was carried out for 4 h. Thionyl chloride was evaporated under reduced pressure, and anhydrous dichloromethane (15 ml) was added. The solvent was evaporated under reduced pressure, and anhydrous dichloromethane (60 ml) was added to the residue to obtain a solution for the next step.
2) At −50° C., paclitaxel (1.6 g) and 4-N,N-dimethylaminopyridine (1.4 g) were added to dichloromethane (DCM, 150 ml), and the mixture was stirred to dissolve. The solution of 4-N,N-dimethylamino-2(R)-fluorobutyryl chloride hydrochloride in dichloromethane prepared in step 1) was slowly added dropwise, and the reaction was detected by HPLC. Upon completion of the reaction, the DCM layer was washed with a saturated aqueous sodium chloride solution (the pH of which was adjusted to about 3.0 with hydrochloric acid), dried over anhydrous sodium sulfate, and filtered. DCM was removed by rotary evaporation, and the residue was crystallized from acetone and methyl tert-butyl ether to obtain a white solid (yield: 50%).
ESI-MS m/z [M+H]+: 985.4.
1H-NMR (400 MHz, DMSO): δ 7.86 (m, 2H), 7.79 (m, 2H), 7.68 (t, 1H), 7.59 (m, 3H), 7.48 (t, 2H), 7.38 (m, 2H), 7.25 (m, 3H), 6.18 (s, 1H), 6.15 (s, 1H), 5.79 (s, 1H), 4.98 (t, 2H), 4.85 (d, J=6.24 Hz, 1H), 4.26 (d, 1H), 3.72 (d, J=7.62 Hz, 1H), 3.64 (t, 1H), 2.89 (t, 2H), 2.54 (s, 1H), 2.26 (t, 2H), 2.18 (m, 13H), 2.01 (s, 3H), 1.95 (m, 2H), 1.86 (m, 1H), 1.75 (m, 1H), 1.68 (m, 1H), 1.52 (s, 6H), 1.41 (s, 3H).
13C-NMR (600 MHz, DMSO): δ=203.64, 171.32, 170.65, 169.63, 168.96, 165.98, 165.86, 140.36, 140.21, 137.12, 134.65, 134.12, 133.98, 133.89, 133.21, 130.65, 129.94, 128.98, 128.54, 128.43, 128.12, 85.12, 81.45, 80.65, 78.23, 76.58, 76.46, 76.45, 74.98, 72.56, 70.23, 59.64, 54.87, 53.03, 48.65, 43.45, 42.26, 42.15, 41.96, 38.96, 34.45, 28.20, 23.97, 21.65, 21.01, 11.23.
Example 11
2′-O-[4-N,N-dimethylamino-2(R)-fluorobutyryl]paclitaxel methanesulfonate (compound C1)
The hydrochloride salt prepared in Example 10 was dissolved in DCM (25 ml), and the solution was washed with a solution of sodium methanesulfonate (above 1 mol/L, pH=3, the pH value was adjusted with methanesulfonic acid) (15 ml*4). The organic layer was dried over anhydrous sodium sulfate and filtered, and the solvent was removed through evaporation under reduced pressure. The residue was crystallized from acetone and methyl tert-butyl ether to give a white solid (yield: 53%).
Example 12
2′-O-[4-N,N-dimethylamino-2(R,S)-fluorobutyryl]paclitaxel hydrochloride (compound 03)
At −50° C., paclitaxel (2 g) and 4-N,N-dimethylaminopyridine (1.9 g) were added to dichloromethane (DCM, 150 ml), and the mixture was stirred to dissolve. The reaction was detected by HPLC. A 3% solution of 4-N,N-dimethylamino-2(R,S)-fluorobutyryl chloride hydrochloride (prepared according to Example 10.1, using 4-N,N-dimethylamino-2(R,S)-fluorobutyric acid hydrochloride as a starting material) in dichloromethane was slowly added dropwise, and the reaction was detected by HPLC. Upon completion of the reaction, the DCM layer was washed with a saturated aqueous sodium chloride solution (the pH of which was adjusted to about 3.0 with hydrochloric acid), dried over anhydrous sodium sulfate, and filtered, and DCM was removed by rotary evaporation. The residue was crystallized from acetone and methyl tert-butyl ether to give a white solid (yield: 48%).
ESI-MS m/z [M+H]+: 985.4.
Example 13
2′-O-[4-N,N-dimethylamino-2(R,S)-difluoromethylbutyryl]paclitaxel citrate (C92)
At −50° C., 4-(N,N-dimethyl)amino-2(R,S)-difluoromethylbutyric acid hydrochloride (2.0 g), 4-N,N-dimethylaminopyridine (1.8 g), and dicyclohexylcarbodiimide (2.1 g) were added to acetonitrile (150 ml). Paclitaxel (1.6 g) was added, and the reaction was carried out for 18 hours. Upon completion of the reaction, the solvent was removed by evaporation under reduced pressure, and the residue was dissolved in dichloromethane (100 ml). The DCM layer was washed with a saturated aqueous sodium chloride solution (the pH of which was adjusted to about 3.0 with hydrochloric acid), dried over anhydrous sodium sulfate, and filtered. DCM was removed by rotary evaporation, and the residue was dissolved in a little acetone. Isopropyl ether was added for crystallization, and 2′-O-[4-N,N-dimethylamino-2(R,S)-difluoromethylbutyryl]paclitaxel hydrochloride (1.2 g) was obtained. The hydrochloride salt was dissolved in dichloromethane (100 ml). The dichloromethane layer was washed with an aqueous solution (pH=3) of citric acid and sodium citrate, dried over anhydrous sodium sulfate, and filtered. The solvent was removed by rotary evaporation, and 2′-O-[4-N,N-dimethylamino-2(R,S)-difluoromethylbutyryl] paclitaxel citrate (1.05 g) was obtained.
ESI-MS m/z [M+H]+: 1017.54.
B-2. Preparation of Docetaxel Derivatives
Example 14
2′-O-[4-N,N-dimethylamino-2(R)-fluorobutyryl]docetaxel hydrochloride (compound 04)
At −15° C., docetaxel (2.1 g) and 4-N,N-dimethylaminopyridine (1.8 g) were added to dichloromethane (DCM, 150 ml), and the mixture was stirred to dissolve. A solution of 4-N,N-dimethylamino-2(R)-fluorobutyryl chloride hydrochloride (prepared according to Example 10.1, using 4-N,N-dimethylamino-2(R)-fluorobutyric acid hydrochloride as a starting material) in dichloromethane was slowly added dropwise, and the reaction was monitored by HPLC. Upon completion of the reaction, the DCM layer was washed with a saturated aqueous sodium chloride solution (the pH of which was adjusted to about 3.0 with hydrochloric acid), dried over anhydrous sodium sulfate, and filtered. DCM was removed by rotary evaporation, and the residue was crystallized from acetone and methyl tert-butyl ether to give a white solid (yield: 51%).
ESI-MS m/z [M+H]+: 939.54.
1H-NMR (400 MHz, DMSO): δ 7.93 (dd, 2H), 7.64 (dt, 1H), 7.40 (m, 2H), 7.19 (d, J=6.42 Hz, 2H), 5.82 (s, 3H), 5.38 (m, 1H), 5.33 (m, 1H), 5.23 (d, J=7.64 Hz, 2H), 5.13 (m, 1H), 5.00 (d, J=5.23 Hz 1H), 4.89 (m, 1H), 4.44 (s, 1H), 4.00 (s, 3H), 3.63 (d, J=7.17 Hz, 2H), 2.59 (dd, 1H), 2.45 (s, 1H), 2.43 (m, 3H), 2.14 (m, 6H), 2.06 (m, 5H), 1.88 (s, 3H), 1.61 (m, 1H), 1.33 (s, 1H), 1.14 (m, 7H), 0.97 (s, 15H), 0.82 (t, 3H).
13C-NMR (600 MHz, DMSO): δ 209.458, 169.376, 168.425, 167.682, 167.513, 166.869, 165.229, 155.167, 137.026, 136.896, 135.884, 133.171, 131.278, 129.960, 129.539, 128.542, 128.044, 127.201, 86.406, 85.187, 83.701, 80.367, 79.033, 78.811, 78.596, 76.788, 75.416, 74.703, 73.715, 71.883, 70.710, 67.308, 57.007, 54.708, 51.949, 45.925, 42.867, 42.515, 42.086, 39.909, 39.771, 39.625, 39.488, 39.350, 39.212, 39.074, 38.077, 36.391, 34.675, 29.777, 28.980, 28.337, 28.061, 26.566, 26.436, 26.298, 23.209, 22.405, 20.627, 13.806, 13.614, 10.717, 9667.
Example 15
2′-O-[4-N,N-dimethylamino-2(R)-fluorobutyryl]docetaxel methanesulfonate (compound C4)
The hydrochloride salt prepared in Example 14 was dissolved in DCM (25 ml), which was washed with a solution of sodium methanesulfonate (above 1 mol/L, pH=3, the pH value was adjusted with methanesulfonic acid) (15 ml×4) The organic layer was dried over anhydrous sodium sulfate, and filtered. The solvent was removed through evaporation under reduced pressure, and the residue was crystallized from acetone and methyl tert-butyl ether to give a white solid (yield: 47%).
Example 16
2′-O-[4-amino-2(R,S)-trifluoromethylbutyryl]docetaxel methanesulfonate (C59)
At −60° C., docetaxel (1.4 g) and 4-N,N-dimethylpyridine (1.2 g) were added to dichloromethane (150 ml), and the mixture was stirred to dissolve. The reaction was detected by HPLC. A 5% solution of 4-N,N-dibenzylamino-2(R,S)-trifluoromethylbutyryl chloride hydrochloride (prepared according to Example 10.1, using 4-N,N-dibenzylamino-2(R,S)-trifluoromethylbutyric acid hydrochloride as a starting material) in dichloromethane was slowly added dropwise, and a solid (1.25 g) was obtained according to the procedures in Example 10. The obtained solid was hydrogenolysis under the catalysis of Pd/C to remove benzyl, and a solid (1.1 g) was obtained, which was dissolved in dichloromethane (100 ml). The dichloromethane layer was washed with an aqueous solution (pH=3) of methanesulfonic acid and sodium methanesulfonate, dried over anhydrous sodium sulfate, and filtered. The solvent was removed by rotary evaporation, and 2′-O-[4-amino-2(R,S)-trifluoromethylbutyryl]docetaxel methanesulfonate (950 mg) was obtained.
ESI-MS m/z [M+H]+: 961.38.
B-3. Preparation of Cabazitaxel Derivatives
Example 17
2′-O-[4-N,N-dimethylamino-2(R,S)-fluorobutyryl]cabazitaxel hydrochloride (compound 09)
At −30° C., cabazitaxel (1.8 g) and 4-N,N-dimethylaminopyridine (1.7 g) were added to dichloromethane (DCM, 150 ml), and the mixture was stirred to dissolve. The reaction was detected by HPLC. A solution of 4-N,N-diethylamino-2(R,S)-fluorobutyryl chloride hydrochloride in dichloromethane was slowly added dropwise. A solid (1.4 g) was obtained according to the procedures in Example 4.
ESI-MS m/z [M+H]+: 967.62.
1H-NMR (400 MHz, DMSO): δ 7.93 (dd, 2H), 7.64 (dt, 1H), 7.40 (m, 2H), 7.19 (d, J=6.42 Hz, 2H), 5.82 (s, 3H), 5.38 (m, 1H), 5.33 (m, 1H), 5.23 (d, J=7.64 Hz, 2H), 5.13 (m, 1H), 5.00 (d, J==5.23 Hz, 1H), 4.89 (m, 1H), 4.44 (s, 1H), 4.00 (s, 3H), 3.63 (d, J=7.17 Hz, 2H), 3.44 (s, 3H), 3.58 (s, 3H), 2.59 (dd, 1H), 2.45 (s, 1H), 2.43 (m, 3H), 2.14 (m, 6H), 2.06 (m, 5H), 1.88 (s, 3H), 1.61 (m, 1H), 1.33 (s, 1H), 1.14 (m, 7H), 0.97 (s, 15H), 0.82 (t, 3H).
13C-NMR (600 MHz, DMSO): δ 209.45, 169.37, 168.42, 167.68, 167.51, 166.87, 165.23, 155.17, 137.02, 136.89, 135.88, 133.17, 131.28, 129.96, 129.54, 128.54, 128.04, 127.20, 86.40, 85.18, 83.70, 80.36, 79.03, 78.81, 78.59, 76.78, 75.42, 74.70, 73.72, 71.88, 70.71, 67.31, 57.01, 56.48, 54.71, 51.95, 45 93, 42.87, 42.52, 42.08, 39.91, 39.77, 39.63, 39.48, 39.35, 39.21, 39.07, 38.08, 36.39, 34.68, 29.78, 28.98, 28.34, 28.06, 26.57, 26.44, 26.29, 23.21, 22.41, 20.63, 13.81, 13.61, 10.72, 9.67.
B-4. Preparation of Larotaxel Derivatives
Example 18
2′-O-[4-N,N-dimethylamino-2(R)-fluorobutyryl]larotaxel hydrochloride (compound C39)
At −20° C., larotaxel (1.3 g) and 4-N,N-dimethylaminopyridine (1.3 g) were added to dichloromethane (DCM, 150 ml), and the mixture was stirred to dissolve. The reaction was detected by HPLC. A solution of 4-N,N-dimethylamino-2(R)-fluorobutyryl chloride hydrochloride in dichloromethane was slowly added dropwise, and a solid (0.9 g) was obtained according to the procedures in Example 10.
ESI-MS m/z [M+H]+: 963.40.
B-5. Preparation of tesetaxel derivatives
Example 19
2′-O-[4-N,N-dimethylamino-2(R)-fluorobutyryl]tesetaxel hydrochloride (compound C43)
At −40° C., tesetaxel (1 g) and 4-N,N-dimethylaminopyridine (0.8 g) were added to dichloromethane (DCM, 100 ml), and the mixture was stirred to dissolve. The reaction was detected by HPLC. A solution of 4-N,N-dimethylamino-2(R)-fluorobutyryl chloride hydrochloride in dichloromethane was slowly added dropwise, and the reaction was carried out according to the procedures in Example 4. Upon completion of the reaction, the DCM layer was washed with a saturated aqueous sodium chloride solution (the pH was adjusted to about 6.0 with hydrochloric acid), dried over anhydrous sodium sulfate, and filtered. DCM was removed by rotary evaporation, and the residue was crystallized from acetone and methyl tert-butyl ether to give a white solid (0.7 g).
ESI-MS m/z [M+H]+: 1013.52.
The inventors also prepared the following compounds according to the methods of the above examples:

    2′-O-[4-N,N-dimethylamino-2(S)-fluorobutyryl]paclitaxel hydrochloride (Compound No. 02): ESI-MS m/z [M+H]+ 985.39
    2′-O-[4-N,N-dimethylamino-2(S)-fluorobutyryl]docetaxel hydrochloride (Compound No. 05): ESI-MS m/z [M+H]+ 939.55
    2′-O-[4-N,N-dimethylamino-2(R,S)-fluorobutyryl]docetaxel hydrochloride (Compound No. 06): ESI-MS m/z [M+H]+ 939.57
    2′-O-[4-N,N-dimethylamino-2(R)-fluorobutyryl]cabazitaxel hydrochloride (Compound No. 07): ESI-MS m/z [M+H]+ 967.59
    2′-O-[4-N,N-dimethylamino-2(S)-fluorobutyryl]cabazitaxel hydrochloride (Compound No. 08): ESI-MS m/z [M+H]+ 967.62
    2′-O-[4-amino-2(R)-difluoromethylbutyryl]cabazitaxel sodium bisulfate salt (Compound No. 10): ESI-MS m/z [M+H]+ 971.41
    2′-O-[4-N,N-diethylamino-2-methyl-2(R)-2-trifluoromethylbutyryl]cabazitaxel methanesulfonate (Compound No. 11): ESI-MS m/z [M+H]+ 1059.49
    2′-O-[4-N-methyl-N-ethylamino-2(R)-2-difluoroethylvaleryl]docetaxel hydrochloride (Compound No. 12): ESI-MS m/z [M+H]+ 1013.45
    2′-O-[4-N,N-dimethylamino-2(R)-fluorovaleryl]docetaxel methanesulfonate (Compound No. 13): ESI-MS m/z [M+H]+ 953.43
    2′-O-[4-(aziridin-1-yl)amino-2,2-difluoroheptanoyl]docetaxel methanesulfonate (Compound No. 14): ESI-MS m/z [M+H]+ 997.37
    2′-O-[4-N-methyl-N-cyclopentylamino-2-trifluoromethyl-2-fluorooctanoyl]paclitaxel methanesulfonate (Compound No. 15): ESI-MS m/z [M+H]+ 1163.41
    2′-O-[4-N,N-dimethylamino-2(R)-fluorovaleryl]larotaxel methanesulfonate (Compound No. 16): ESI-MS m/z [M+H]+ 977.29
    2′-O-[4-N,N-dimethylamino-2(R)-fluorovaleryl]tesetaxel methanesulfonate (Compound No. 17): ESI-MS m/z [M+H]+ 1027.39
    2′-O-[4-N,N-dimethylamino-2(S)-fluorobutyryl]paclitaxel methanesulfonate (Compound No. C2): ESI-MS m/z [M+H]+ 985.37
    2′-O-[4-N,N-dimethylamino-2(R,S)-fluorobutyryl]paclitaxel methanesulfonate (Compound No. C3): ESI-MS m/z [M+H]+ 985.41
    2′-O-[4-N,N-dimethylamino-2(S)-fluorobutyryl]docetaxel methanesulfonate (Compound No. C5): ESI-MS m/z [M+H]+ 939.55
    2′-O-[4-N,N-dimethylamino-2(R,S)-fluorobutyryl]docetaxel methanesulfonate (Compound No. C6): ESI-MS m/z [M+H]+ 939.56
    2′-O-[4-N,N-dimethylamino-2(R)-fluorobutyryl]cabazitaxel methanesulfonate (Compound No. C7): ESI-MS m/z [M+H]+ 967.58
    2′-O-[4-N,N-dimethylamino-2(S)-fluorobutyryl]cabazitaxel methanesulfonate (Compound No. C8): ESI-MS m/z [M+H]+ 967.54
    2′-O-[4-N,N-dimethylamino-2(R,S)-fluorobutyryl]cabazitaxel methanesulfonate (Compound No. C9): ESI-MS m/z [M+H]+ 967.62
    2′-O-[4-amino-2(R)-difluoromethylbutyryl]cabazitaxel sulfate (Compound No. C10): ESI-MS m/z [M+H]+ 971.39
    2′-O-[4-N,N-diethylamino-2-methyl-2(R)-2-trifluoromethylbutyryl]cabazitaxel hydrochloride (Compound No. C11): ESI-MS m/z [M+H]+ 1059.54
    2′-O-[4-N-methyl-N-ethylamino-2(R)-2-difluoroethylvaleryl]docetaxel methanesulfonate (Compound No. C12): ESI-MS m/z [M+H]+ 1013.43
    2′-O-[4-N,N-dimethylamino-2(R)-fluorovaleryl]docetaxel hydrochloride (Compound No. C13): ESI-MS m/z [M+H]+ 953.41
    2′-O-[4-(aziridin-1-yl)amino-2,2-difluoroheptanoyl]docetaxel hydrochloride (Compound No. C14): ESI-MS m/z [M+H]+ 997.39
    2′-O-[4-N-methyl-N-cyclopentylamino-2-trifluoromethyl-2-fluorooctanoyl]paclitaxel hydrochloride (Compound No. C15): ESI-MS m/z [M+H]+ 1163.38
    2′-O-[4-N,N-dimethylamino-2(R)-fluorovaleryl]larotaxel hydrochloride (Compound No. C16): ESI-MS m/z [M+H]+ 977.30
    2′-O-[4-N,N-dimethylamino-2(R)-fluorovaleryl]tesetaxel hydrochloride (Compound No. C17): ESI-MS m/z [M+H]+ 1027.36
    2′-O-[4-N-methyl-N-ethylamino-2(R)-fluorobutyryl]paclitaxel methanesulfonate (Compound No. C18): ESI-MS m/z [M+H]+ 999.42
    2′-O-[4-N,N-diethylamino-2(S)-fluorobutyryl]paclitaxel fumarate (Compound No. C19): ESI-MS m/z [M+H]+ 1013.41
    2′-O-[4-N-methyl-N-isopropylamino-2(R,S)-fluorobutyryl]paclitaxel hydrochloride (Compound No. C20): ESI-MS m/z [M+H]+ 1013.43
    2′-O-[4-N,N-dimethylamino-2(R,S)-2-trifluoromethylbutyryl]paclitaxel p-toluene sulfonate (Compound No. C21): ESI-MS m/z [M+H]+ 1035.38
    2′-O-[4-N,N-dimethylamino-2(R,S)-2-difluoromethylbutyryl]paclitaxel hydrochloride (Compound No. C22): ESI-MS m/z [M+H]+ 1017.23
    2′-O-[4-N,N-dimethylamino-2(R)-fluorovaleryl]paclitaxel maleate (Compound No. C23): ESI-MS m/z [M+H]+ 999.34
    2′-O-[4-N,N-dimethylamino-2(S)-2-difluoromethylhexanoyl]paclitaxel sulfate (Compound No. C24): ESI-MS m/z [M+H]+ 1045.44
    2′-O-[4-N-methyl-N-ethylamino-2(R)-fluorobutyryl]docetaxel methanesulfonate (Compound No. C25): ESI-MS m/z [M+H]+ 953.44
    2′-O-[4-N,N-diethylamino-2(S)-fluorobutyryl]docetaxel maleate (Compound No. C26): ESI-MS m/z [M+H]+ 967.45
    2′-O-[4-N-methyl-N-isopropylamino-2(R)-fluorobutyryl]docetaxel hydrochloride (Compound No. C27): ESI-MS m/z [M+H]+ 967.42
    2′-O-[4-N,N-dimethylamino-2(R)-2-trifluoromethylbutyryl]docetaxel methanesulfonate (Compound No. C28): ESI-MS m/z [M+H]+ 989.38
    2′-O-[4-N,N-dimethylamino-2(R,S)-2-difluoromethylbutyryl]docetaxel hydrochloride (Compound No. C29): ESI-MS m/z [M+H]+ 971.35
    2′-O-[4-N,N-dimethylamino-2(R,S)-fluorovaleryl]docetaxel methanesulfonate (Compound No. C30): ESI-MS m/z [M+H]+ 953.47
    2′-O-[4-N,N-dimethylamino-2(S)-fluorohexanoyl]docetaxel sulfate (Compound No. C31): ESI-MS m/z [M+H]+ 967.41
    2′-O-[4-N-methyl-N-ethylamino-2(R)-fluorobutyryl]cabazitaxel maleate (Compound No. C32): ESI-MS m/z [M+H]+ 981.35
    2′-O-[4-N,N-diethylamino-2(R)-fluorobutyryl]cabazitaxel methanesulfonate (Compound No. C33): ESI-MS m/z [M+H]+ 995.51
    2′-O-[4-N-methyl-N-isopropylamino-2(R)-fluorobutyryl]cabazitaxel hydrochloride (Compound No. C34): ESI-MS m/z [M+H]+ 995.47
    2′-O-[4-N,N-dimethylamino-2(R)-2-trifluoromethylbutyryl]cabazitaxel methanesulfonate (Compound No. C35): ESI-MS m/z [M+H]+ 1017.52
    2′-O-[4-N,N-dimethylamino-2(R)-2-difluoromethylbutyryl]cabazitaxel p-toluene sulfonate (Compound No. C36): ESI-MS m/z [M+H]+ 999.46
    2′-O-[4-N,N-dimethylamino-2(R)-fluorovaleryl]cabazitaxel hydrochloride (Compound No. C37): ESI-MS m/z [M+H]+ 981.33
    2′-O-[4-N,N-dimethylamino-2(R)-fluorohexanoyl]cabazitaxel sulfate (Compound No. C38): ESI-MS m/z [M+H]+ 995.34
    2′-O-[4-N-methyl-N-ethylamino-2(R,S)-fluorobutyryl]larotaxel hydrochloride (Compound No. C40): ESI-MS m/z [M+H]+ 977.54
    2′-O-[4-N-methyl-N-isopropylamino-2(R,S)-2-trifluoromethylbutyryl]larotaxel maleate (Compound No. C41): ESI-MS m/z [M+H]+ 1041.35
    2′-O-[4-N,N-dimethylamino-2(S)-fluorovaleryl]larotaxel phosphate (Compound No. C42): ESI-MS m/z [M+H]+ 977.65
    2′-O-[4-N,N-diethylamino-2(S)-fluorobutyryl]tesetaxel hydrochloride (Compound No. C44): ESI-MS m/z [M+H]+ 1041.52
    2′-O-[4-N-methyl-N-isopropylamino-2(R,S)-2-trifluoromethylbutyryl]tesetaxel succinate (Compound No. C45): ESI-MS m/z [M+H]+ 1091.51
    2′-O-[4-(pyrrolidin-1-yl)amino-2(R,S)-fluorovaleryl]tesetaxel sulfate (Compound No. C46): ESI-MS m/z [M+H]+ 1053.48
    2′-O-[4-N,N-diethylamino-2(R)-fluorobutyryl]paclitaxel hydrochloride (Compound No. C51): ESI-MS m/z [M+H]+ 1013.35
    2′-O-[3-N,N-dimethylamino-2(R,S)-fluoropropionyl]paclitaxel hydrochloride (Compound No. C52): ESI-MS m/z [M+H]+ 971.28
    2′-O-[4-N-benzylamino-2(S)-fluorobutyryl]docetaxel hydrochloride (Compound No. C62): ESI-MS m/z [M+H]+ 1001.36
    2′-O-[3-(N,N-diethyl)amino-2(R,S)-trifluoromethylpropionyl]docetaxel hydrochloride (Compound No. C63): ESI-MS m/z [M+H]+ 1003.34
    2′-O-[4-(N-methyl)amino-2(R)-trifluoromethylbutyryl]paclitaxel hydrochloride (Compound No. C65): ESI-MS m/z [M+H]+ 1021.24
    2′-O-[3-(N-isopropyl)amino-2(R,S)-difluoromethylpropionyl]cabazitaxel hydrochloride (Compound No. C66): ESI-MS m/z [M+H]+ 999.38
    2′-O-[5-(N,N-dimethyl)amino-2(R)-fluorovaleryl]paclitaxel hydrochloride (Compound No. C74): ESI-MS m/z [M+H]+ 999.33
    2′-O-[4-(N,N-dimethyl)amino-2(S)-trifluoromethylbutyryl]cabazitaxel hydrochloride (Compound No. C75): ESI-MS m/z [M+H]+ 1017.41
    2′-O-[4-(N-isopropyl)amino-2(R,S)-trifluoromethylbutyryl]paclitaxel hydrochloride (Compound No. C76): ESI-MS m/z [M+H]+ 1049.37
    2′-O-[5-N,N-dimethylamino-2(S)-trifluoromethylvaleryl]docetaxel hydrochloride (Compound No. C79): ESI-MS m/z [M+H]+ 1003.31
    2′-O-[4-N-benzylamino-2-methyl-2(R,S)-fluorobutyryl]paclitaxel hydrochloride (Compound No. C80): ESI-MS m/z [M+H]+ 1061.32
    2′-O-[4-N,N-dimethylamino-2(R)-difluoromethylbutyryl]docetaxel hydrochloride (Compound No. C87): ESI-MS m/z [M+H]+ 971.29
    2′-O-[3-cyclopentylamino-2-ethyl-2(R,S)-trifluoromethylpropionyl]docetaxel hydrochloride (Compound No. C95): ESI-MS m/z [M+H]+ 1001.31
    2′-O-[5-N-benzylamino-2-benzyl-2(R)-difluoromethylvaleryl]cabazitaxel hydrochloride (Compound No. C97): ESI-MS m/z [M+H]+ 1075.42
    2′-O-[4-(4-piperidin-1-yl)-2(S)-trifluoromethylbutyryl]cabazitaxel hydrochloride (Compound No. C99): ESI-MS m/z [M+H]+ 1057.35
    2′-O-[4-N,N-dimethylamino-2(S)-fluorobutyryl]larotaxel hydrochloride (Compound No. C102): ESI-MS m/z [M+H]+ 963.34
    2′-O-[4-N,N-dimethylamino-2(R)-fluorobutyryl]tesetaxel hydrochloride (Compound No. C107): ESI-MS m/z [M+H]+ 1013.41.


The inventors provide the following experimental examples to show the surprising and unexpected beneficial effects of the water soluble taxane derivatives of the present invention.
EXPERIMENTAL EXAMPLES
Experimental Example 1.1. Solubility in Physiological Saline











TABLE 1







Solubility of the water soluble taxane derivatives in physiological saline



















Compd.
Type of the










Solubility


No.
parent ring
n
R1
R2
R3
R4
R5
X
Y
C*
A
(mg/ml)






















01
I-1
2
H
Ac
Ph
Me
Me
H
F
R
HCl
1.1


02
I-1
2
H
Ac
Ph
Me
Me
H
F
S
HCl
1.2


03
I-1
2
H
Ac
Ph
Me
Me
H
F
R, S
HCl
1.1


04
I-1
2
H
H
t-BuO
Me
Me
H
F
R
HCl
2.8


05
I-1
2
H
H
t-BuO
Me
Me
H
F
S
HCl
2.5


06
I-1
2
H
H
t-BuO
Me
Me
H
F
R, S
HCl
2.6


07
I-1
2
Me
Me
t-BuO
Me
Me
H
F
R
HCl
1.5


08
I-1
2
Me
Me
t-BuO
Me
Me
H
F
S
HCl
1.3


09
I-1
2
Me
Me
t-BuO
Me
Me
H
F
R, S
HCl
1.3


10
I-1
2
Me
Me
t-BuO
H
H
H
CHF2
R
NaHSO4
1.6


11
I-1
2
Me
Me
t-BuO
Et
Et
CH3
CF3
R
MeSO3H
11.2


12
I-1
3
H
H
t-BuO
Me
Et
H
CF2CH3
R
HCl
1.2


13
I-1
3
H
H
t-BuO
Me
Me
H
F
R
MeSO3H
15.6


16
I-2
3

Ac
t-BuO
Me
Me
H
F
R
MeSO3H
11.1


17
I-3
3


t-BuO
Me
Me
H
F
R
MeSO3H
12.3


C1
I-1
2
H
Ac
Ph
Me
Me
H
F
R
MeSO3H
10.5


C2
I-1
2
H
Ac
Ph
Me
Me
H
F
S
MeSO3H
10.4


C3
I-1
2
H
Ac
Ph
Me
Me
H
F
R, S
MeSO3H
10.3


C4
I-1
2
H
H
t-BuO
Me
Me
H
F
R
MeSO3H
12.4


C5
I-1
2
H
H
t-BuO
Me
Me
H
F
S
MeSO3H
12.7


C6
I-1
2
H
H
t-BuO
Me
Me
H
F
R, S
MeSO3H
12.5


C7
I-1
2
Me
Me
t-BuO
Me
Me
H
F
R
MeSO3H
0.9


C8
I-1
2
Me
Me
t-BuO
Me
Me
H
F
S
MeSO3H
0.6


C9
I-1
2
Me
Me
t-BuO
Me
Me
H
F
R, S
MeSO3H
0.4


C10
I-1
2
Me
Me
t-BuO
H
H
H
CHF2
R
H2SO4
0.8


C11
I-1
2
Me
Me
t-BuO
Et
Et
CH3
CF3
R
HCl
1.1


C12
I-1
3
H
H
t-BuO
Me
Et
H
CF2CH3
R
MeSO3H
11.3


C13
I-1
3
H
H
t-BuO
Me
Me
H
F
R
HCl
2.3


C16
I-2
3

Ac
t-BuO
Me
Me
H
F
R
HCl
0.8


C17
I-3
3


t-BuO
Me
Me
H
F
R
HCl
1.0









Experimental Example 2. In Vitro Dissociation in Blood Plasma of Rat
The obtained water soluble taxane derivatives of the following formulae were formulated as 0.2 mg/ml aqueous solutions. 0.1 ml samples were taken from each of the solutions, added respectively to 0.9 ml of fresh blood plasma (anticoagulated with heparin) from SD rats, homogeneously mixed, and placed in a thermostatic water bath at 37° C. for incubation with time being recorded. After 2 min, 5 min and 10 min of incubation, 0.2 ml of solutions were respectively taken from each of the samples, and added to acetonitrile (0.4 ml) cooled to −20° C. for precipitation of protein. The samples were shaken and centrifuged for 10 min (10,000 rpm), and the supernatant was then taken for HPLC analysis. The results are presented in following Table 2:











TABLE 2







Test of in vitro dissociation of the water soluble taxane derivatives in blood plasma of rat




















Type of










Sampling time/Dissociation


Compd.
the parent










percent (%)





















No.
ring
n
R1
R2
R3
R4
R5
X
Y
C* 
A
2 min
5 min
10 min
























01
I-1
2
H
Ac
Ph
Me
Me
H
F
R
HCl
63.5
96.8
100


02
I-1
2
H
Ac
Ph
Me
Me
H
F
S
HCl
59.8
95.6
100


03
I-1
2
H
Ac
Ph
Me
Me
H
F
R, S
HCl
61.8
93.1
100


04
I-1
2
H
H
t-BuO
Me
Me
H
F
R
HCl
58.6
92.6
100


05
I-1
2
H
H
t-BuO
Me
Me
H
F
S
HCl
56.4
91.1
100


06
I-1
2
H
H
t-BuO
Me
Me
H
F
R, S
HCl
56.4
91.1
100


07
I-1
2
Me
Me
t-BuO
Me
Me
H
F
R
HCl
49.5
89.6
100


08
I-1
2
Me
Me
t-BuO
Me
Me
H
F
S
HCl
47.6
87.6
100


09
I-1
2
Me
Me
t-BuO
Me
Me
H
F
R, S
HCl
47.8
89.1
100


10
I-1
2
Me
Me
t-BuO
H
H
H
CHF2
R
NaHSO4
45.2
86.5
96.8


11
I-1
2
Me
Me
t-BuO
Et
Et
CH3
CF3
R
MeSO3H
52.3
89.6
97.2


12
I-1
3
H
H
t-BuO
Me
Et
H
CF2CH3
R
HCl
43.6
75.6
89.6


13
I-1
3
H
H
t-BuO
Me
Me
H
F
R
MeSO3H
45.7
85.2
92.3




















14
I-1
5
H
H
t-BuO
—(CH2)2—
F
F
—
MeSO3H
42.1
55.7
72.5


 





















15
I-1
6
H
Ac
Ph
Me




F
CF3
R, S
MeSO3H
35.0
46.2
57.9


 


16
I-2
3

Ac
t-BuO
Me
Me
H
F
R
MeSO3H
60.2
92.1
100


17
I-3
3


t-BuO
Me
Me
H
F
R
MeSO3H
59.3
93.5
100


C1
I-1
2
H
Ac
Ph
Me
Me
H
F
R
MeSO3H
64.3
97.0
100


C2
I-1
2
H
Ac
Ph
Me
Me
H
F
S
MeSO3H
62.8
96.5
100


C3
I-1
2
H
Ac
Ph
Me
Me
H
F
R, S
MeSO3H
63.7
96.8
100


C4
I-1
2
H
H
t-BuO
Me
Me
H
F
R
MeSO3H
59.2
93.4
100


C5
I-1
2
H
H
t-BuO
Me
Me
H
F
S
MeSO3H
58.9
93.3
100


C6
I-1
2
H
H
t-BuO
Me
Me
H
F
R, S
MeSO3H
59.1
93.3
100


C7
I-1
2
Me
Me
t-BuO
Me
Me
H
F
R
MeSO3H
49.7
88.9
100


C8
I-1
2
Me
Me
t-BuO
Me
Me
H
F
S
MeSO3H
49.4
88.4
100


C9
I-1
2
Me
Me
t-BuO
Me
Me
H
F
R, S
MeSO3H
49.3
88.6
100


C10
I-1
2
Me
Me
t-BuO
H
H
H
CHF2
R
H2SO4
45.6
87.1
97.3


C11
I-1
2
Me
Me
t-BuO
Et
Et
CH3
CF3
R
HCl
51.2
88.4
95.9


C12
I-1
3
H
H
t-BuO
Me
Et
H
CF2CH3
R
MeSO3H
42.8
74.8
88.4


C13
I-1
3
H
H
t-BuO
Me
Me
H
F
R
HCl
44.8
84.8
92.7




















C14
I-1
5
H
H
t-BuO
—(CH2)2—
F
F
—
HCl
41.9
54.4
71.3


 





















C15
I-1
6
H
Ac
Ph
Me




F
CF3
R, S
HCl
34.2
45.3
58.2


 


C16
I-2
3

Ac
t-BuO
Me
Me
H
F
R
HCl
61.1
93.8
100


C17
I-3
3


t-BuO
Me
Me
H
F
R
HCl
60.7
95.5
100









Experimental Example 3. In Vitro Dissociation in Blood Plasma of Rabbit
The obtained water soluble taxane derivatives of the following formulae were formulated as 0.2 mg/ml aqueous solutions. 0.1 ml samples were taken from each of the solutions, added respectively to 0.9 ml of fresh blood plasma (anticoagulated with heparin) from New Zealand white rabbits, homogeneously mixed, and placed m a thermostatic water bath at 37° C. for incubation with time being recorded. After 5 min, 20 min and 60 min of incubation, 0.2 ml of solutions were respectively taken from each of the samples, and added to acetonitrile (0.4 ml) cooled to −20° C. for precipitation of protein. The samples were shaken and centrifuged for 10 min (10.000 rpm), and the supernatant was then taken for HPLC analysis. The results are presented in following Table 3:











TABLE 3







Test of in vitro dissociation of the water soluble taxane derivatives in blood plasma of rabbit































Sampling time/


Compd.
Type of the










dissociation percent (%)





















No.
parent ring
n
R1
R2
R3
R4
R5
X
Y
C*
A
2 min
5 min
10 min
























01
I-1
2
H
Ac
Ph
Me
Me
H
F
R
HCl
76.5
98.3
100


02
I-1
2
H
Ac
Ph
Me
Me
H
F
S
HCl
68.2
97.5
100


03
I-1
2
H
Ac
Ph
Me
Me
H
F
R, S
HCl
75.7
93.5
100


04
I-1
2
H
H
t-BuO
Me
Me
H
F
R
HCl
80.2
93.5
100


05
I-1
2
H
H
t-BuO
Me
Me
H
F
S
HCl
79.3
91.8
100


06
I-1
2
H
H
t-BuO
Me
Me
H
F
R, S
HCl
75.6
98.6
100


07
I-1
2
Me
Me
t-BuO
Me
Me
H
F
R
HCl
79.2
95.5
100


08
I-1
2
Me
Me
t-BuO
Me
Me
H
F
S
HCl
89.9
93.2
100


09
I-1
2
Me
Me
t-BuO
Me
Me
H
F
R, S
HCl
83.2
97.1
100


10
I-1
2
Me
Me
t-BuO
H
H
H
CHF2
R
NaHSO4
38.1
63.2
80.3


11
I-1
2
Me
Me
t-BuO
Et
Et
CH3
CF3
R
MeSO3H
56.2
80.5
92.1


12
I-1
3
H
H
t-BuO
Me
Et
H
CF2CH3
R
HCl
35.2
61.3
73.2


13
I-1
3
H
H
t-BuO
Me
Me
H
F
R
MeSO3H
33.2
55.8
62.1


16
I-2
3

Ac
t-BuO
Me
Me
H
F
R
MeSO3H
63.2
94.1
100


17
I-3
3


t-BuO
Me
Me
H
F
R
MeSO3H
62.7
93.4
100


C1
I-1
2
H
Ac
Ph
Me
Me
H
F
R
MeSO3H
77.2
98.4
100


C2
I-1
2
H
Ac
Ph
Me
Me
H
F
S
MeSO3H
69.7
98.1
100


C3
I-1
2
H
Ac
Ph
Me
Me
H
F
R, S
MeSO3H
76.5
94.6
100


C4
I-1
2
H
H
t-BuO
Me
Me
H
F
R
MeSO3H
81.6
94.8
100


C5
I-1
2
H
H
t-BuO
Me
Me
H
F
S
MeSO3H
81.2
93.1
100


C6
I-1
2
H
H
t-BuO
Me
Me
H
F
R, S
MeSO3H
76.8
97.8
100


C7
I-1
2
Me
Me
t-BuO
Me
Me
H
F
R
MeSO3H
78.4
94.8
100


C8
I-1
2
Me
Me
t-BuO
Me
Me
H
F
S
MeSO3H
85.6
94.5
100


C9
I-1
2
Me
Me
t-BuO
Me
Me
H
F
R, S
MeSO3H
81.2
96.7
100


C10
I-1
2
Me
Me
t-BuO
H
H
H
CHF2
R
H2SO4
48.9
68.4
91.3


C11
I-1
2
Me
Me
t-BuO
Et
Et
CH3
CF3
R
HCl
61.2
84.5
96.3


C12
I-1
3
H
H
t-BuO
Me
Et
H
CF2CH3
R
MeSO3H
44.6
72.1
89.6


C13
I-1
3
H
H
t-BuO
Me
Me
H
F
R
HCl
46.2
78.9
93.4


C16
I-2
3

Ac
t-BuO
Me
Me
H
F
R
HCl
68.2
94.4
100


C17
I-3
3


t-BuO
Me
Me
H
F
R
HCl
69.7
96.8
100









Experimental Example 4. In Vivo Metabolic Test of the Water Soluble Paclitaxel Derivatives in Rats
4.1. Method of Study:
12 SD rats (male, body weight: 200-220 g) were randomly divided into four groups, and intravenously administered with 5 mg/kg of compounds 01, 02, 03 and commercially available paclitaxel in a volume of 5 ml/kg, respectively. Compounds 01, 02 and 03 were formulated with 5% dextrose injection (pH=5), and paclitaxel is in the form of a commercially available injection. After 5 min of administration, 0.3 ml of venous blood was taken from the retrobulbar venous plexus of the rats, placed in heparinized tubes, and centrifuged at 11.000 rpm for 5 min. Blood plasma was separated, and the concentrations of the compounds in the blood plasma were determined by liquid chromatography-mass spectrometry.
4.2. Results:
After intravenous injection of compounds 01, 02, and 03, compounds 01, 02, and 03 cannot be detected in the blood plasma, and only paclitaxel can be detected.
The mean concentrations (ng/ml) of paclitaxel in the blood plasma of animals from each group were 1789, 1637, 1825, and 1793, respectively.
Experimental Example 5. In Vivo Metabolic Test of the Water Soluble Docetaxel Derivatives in Rats
5.1. Method of Study:
12 SD rats (male, body weight: 200-220) g) were randomly divided into four groups, and intravenously administered with 5 mg/kg of compounds 04, 05, 06 and commercially available docetaxel m a volume of 5 ml/kg, respectively. Compounds 04, 05 and 06 were formulated with 5% dextrose injection (pH=5), and docetaxel is in the form of a commercially available injection. After 5 min of administration, 0.3 ml of venous blood was taken from the retrobulbar venous plexus of the rats, placed in heparinized tubes, and centrifuged at 11,000 rpm for 5 min. Plasma was separated, and the concentrations of the compounds in the plasma were determined by liquid chromatography □-mass spectrometry.
5.2. Results:
After intravenous injection of compounds 04, 05, and 06, compounds 04, 05, and 06 cannot be detected in the blood plasma, and only docetaxel can be detected.
The mean concentrations (ng/ml) of docetaxel in the blood plasma of animals from each group were 1506, 1387, 1621, and 769, respectively.
Experimental Example 6. Antitumor Activity Test of the Water Soluble Taxane Derivatives
6.1. Test of the Inhibitory Activity of the Water Soluble Paclitaxel Derivatives on Subcutaneously Xenografted Tumor of Human Ovarian Cancer SK-OV-3 in Nude Mice
Purpose: to evaluate and compare the inhibitory activity of the water soluble paclitaxel derivatives, paclitaxel and Abraxane® on subcutaneously xenografted tumor of human ovarian cancer SK-OV-3 in nude mice.
Dosage Regimen and Experimental Procedures:
Human ovarian cancer SK-OV-3 cells were subcutaneously inoculated to nude mice. After the tumor volume reached 100-150 mm3, the animals were randomly divided into groups (D0), and administered with the water soluble paclitaxel derivatives of the following formula, paclitaxel, and Abraxane®, respectively once per day for 5 days. The dosage and dosage regimen are shown in following table 4. The tumor volume was measured 2-3 times per week, the animals were weighed and the data were recorded until day 22 (D22) after grouping.
The tumor volume (V) was calculated according to the equation: V=1⁄2×a×b2, wherein a and b represent the length and the width, respectively.
T/C (%)=(T−T0)/(C−C0)×100, wherein T and C represent the tumor volume at the end of the experiment, and T0 and C0 represent the tumor volume at the beginning of the experiment.
The antitumor activity data are shown in following Table 4:











TABLE 4





The inhibitory activity of the water soluble paclitaxel derivatives, paclitaxel and Abraxane ® on


subcutaneously xenografted tumor of human ovarian cancer SK-OV-3 in nude mice





























Time and












Compd.

route of












No.
Dosage
administration
n
R1
R2
R3
R4
R5
X
Y
C*
A





the control
—
D0-4 IV
—
—
—
—
—
—
—
—
—
—


group














paclitaxel
16 mg/kg
D0-4 IV
—
—
—
—
—
—
—
—
—
—


Abraxane ®
24 mg/kg
D0-4 IV
—
—
—
—
—
—
—
—
—
—


C1
24 mg/kg
D0-4 IV
2
H
Ac
Ph
Me
Me
H
F
R
MeSO3H


C2
24 mg/kg
D0-4 IV
2
H
Ac
Ph
Me
Me
H
F
S
MeSO3H


C3
24 mg/kg
D0-4 IV
2
H
Ac
Ph
Me
Me
H
F
R, S
MeSO3H


C18
24 mg/kg
D0-4 IV
2
H
Ac
Ph
Et
Me
H
F
R
MeSO3H


C19
24 mg/kg
D0-4 IV
2
H
Ac
Ph
Et
Et
H
F
S
fumaric acid


C20
24 mg/kg
D0-4 IV
2
H
Ac
Ph
Me
i-Pro
H
F
R, S
HCl


C21
24 mg/kg
D0-4 IV
2
H
Ac
Ph
Me
Me
H
CF3
R, S
p-toluene














sulfonic acid


C22
24 mg/kg
D0-4 IV
2
H
Ac
Ph
Me
Me
H
CHF2
R, S
HCl


C23
24 mg/kg
D0-4 IV
3
H
Ac
Ph
Me
Me
H
F
R
maleic acid


C24
24 mg/kg
D0-4 IV
4
H
Ac
Ph
Me
Me
H
CHF2
S
sulfuric acid












Anti-tumor activity data


















Tumor
Body




Tumor
Tumor

inhibitory
weight



Compd.
volume
volume
T/C
percent
change



No.
(mm3) (D0)
(mm3) (D22)
(%) D22
(%)Δ D22
(%) D4






the control
134.6 ± 18.3
980.4 ± 271.9
—
—
−11



group








paclitaxel
134.6 ± 19.7
53.7 ± 11.2
−60
160
−4.8



Abraxane ®
126.0 ± 10.7
53.8 ± 13.9
−57
157
−8.2



C1
133.6 ± 9.5 
49.8 ± 10.4
−63
163
−5.9



C2
134.3 ± 10.4
51.2 ± 11.2
−61
158
−5.8



C3
133.4 ± 10.4
50.6 ± 9.7 
−64
165
−6.0



C18
133.6 ± 9.4 
55.7 ± 10.6
−53
137
−7.0



C19
133.6 ± 9.4 
55.7 ± 10.1
−44
114
−8.1



C20
127.9 ± 11.4
60.4 ± 12.3
−38
98
−10.3



C21
134.9 ± 11.4
57.3 ± 11.4
−48
124
−8.9



C22
133.4 ± 12.6
61.2 ± 13.5
−35
135
−8.4



C23
135.8 ± 14.7
70.8 ± 16.8
−23
60
−9.4



C24
133.8 ± 12.2
79.8 ± 12.8
−14
36
−11.6





D0: the time of administration for the first time;


ΔP(D22) = 0.000, compared to the control, using Student's t-test.


The numbers of mice at the beginning of the experiment: n = 10 in the control group, and n = 6 in the administered group.


C* is the configuration of the chiral carbon in the side chain of an amino acid.


Conclusion: the water soluble paclitaxel derivatives have an inhibitory effect on human ovarian cancer SK-OV-3.






6.2. Test of the Inhibitory Activity of the Water Soluble Docetaxel Derivatives on Subcutaneously Xenografted Tumor of Human Prostate Cancer PC-3 in Nude Mice
Purpose: to evaluate and compare the inhibitory activity of the water soluble docetaxel derivatives and docetaxel on subcutaneously xenografted tumor of human prostate cancer PC-3 in nude mice
Dosage Regimen and Experimental Procedures:
Human prostate cancer PC-3 cells were subcutaneously inoculated into nude mice. After the tumor volume reached 100-150 mm3, the animals were randomly divided into several groups, and administered with the water soluble docetaxel derivatives of the following formula and docetaxel, respectively once on the same day (D0). The dosage and dosage regimen are shown in following table 5. The tumor volume was measured 2-3 times per week, the animals were weighed and the data were recorded until day 20 (D20) after grouping.
The tumor volume (V) was calculated according to the equation: V=1⁄2×a×b2, wherein a and b represent the length and the width, respectively.
T/C (%)=(T−T0)/(C−C0)×100, wherein T and C represent the tumor volume at the end of the experiment; and T0 and C0 represent the tumor volume at the beginning of the experiment.
The antitumor activity data are shown in following Table 5:











TABLE 5





The inhibitory activity of the water soluble docetaxel derivatives and docetaxel


on subcutaneously xenografted tumor of human prostate cancer PC-3 in nude mice.





























Time and












Compd.

route of












No.
Dosage
administration
n
R1
R2
R3
R4
R5
X
Y
C*
A





the control
—
D0 IV
—
—
—
—
—
—
—
—
—
—


group














docetaxel
  14 mg/kg
D0 IV
—
—
—
—
—
—
—
—
—
—


C4
33.6 mg/kg
D0 IV
2
H
H
t-BuO
Me
Me
H
F
R
MeSO3H


C5
33.6 mg/kg
D0 IV
2
H
H
t-BuO
Me
Me
H
F
S
MeSO3H


C6
33.6 mg/kg
D0 IV
2
H
H
t-BuO
Me
Me
H
F
R, S
MeSO3H


C25
33.6 mg/kg
D0 IV
2
H
H
t-BuO
Et
Me
H
F
R
MeSO3H


C26
33.6 mg/kg
D0 IV
2
H
H
t-BuO
Et
Et
H
F
S
maleic acid


C27
33.6 mg/kg
D0 IV
2
H
H
t-BuO
Me
i-Pro
H
F
R
HCl


C28
33.6 mg/kg
D0 IV
2
H
H
t-BuO
Me
Me
H
CF3
R
MeSO3H


C29
33.6 mg/kg
D0 IV
2
H
H
t-BuO
Me
Me
H
CHF2
R, S
HCl


C30
33.6 mg/kg
D0 IV
3
H
H
t-BuO
Me
Me
H
F
R, S
MeSO3H


C31
33.6 mg/kg
D0 IV
4
H
H
t-BuO
Me
Me
H
F
S
sulfuric acid












Anti-tumor activity data


















Tumor





Tumor
Tumor

inhibitory
Weight



Compd.
volume
volume
T/C
percent
change



No.
(mm3) (D0)
(mm3) (D20)
(%) D20
(%)Δ D20
(%) D6






the control
110.5 ± 4.4
870.5 ± 60.5 
—
—
−16



group








docetaxel
111.5 ± 1.3
81.4 ± 25.1
−27
127
−12.3



C4
113.7 ± 2.9
26.6 ± 12.4
−77
177
−12.5



C5
113.4 ± 3.1
26.8 ± 12.6
−76
175
−12.4



C6
113.8 ± 3.0
27.0 ± 12.3
−76
175
−12.6



C25
113.8 ± 3.0
30.4 ± 11.1
−62
143
−13.6



C26
114.4 ± 2.8
32.4 ± 13.5
−52
120
−14.1



C27
116.3 ± 3.7
35.3 ± 14.7
−44
101
−14.4



C28
113.5 ± 2.8
34.5 ± 12.6
−46
105
−13.9



C29
116.7 ± 3.1
36.8 ± 13.4
−37
85
−15.1



C30
118.2 ± 4.8
41.5 ± 14.7
−28
64
−15.4



C31
114.7 ± 3.5
52.4 ± 12.5
−20
45
−15.8





D0: the time of administration for the first time;


ΔP(D20) = 0.000, compared to the control, using Student's t-test.


The numbers of mice at the beginning of the experiment: n = 10 in the control group, and n = 6 in the administered group.


C* is the configuration of the chiral carbon in the side chain of an amino acid.


Conclusion: the water soluble docetaxel derivatives have an inhibitory effect on human prostate cancer PC-3.






6.3. Test of the Inhibitory Activity of the Water Soluble Cabazitaxel Derivatives on to Subcutaneously Xenografted Tumor of Human Prostate Cancer PC-3 in Nude Mice
Purpose: to evaluate and compare the inhibitory activity of the water soluble cabazitaxel derivatives and cabazitaxel on subcutaneously xenografted tumor of human prostate cancer PC-3 in nude mice.
Dosage Regimen and Experimental Procedures:
Human prostate cancer PC-3 cells were subcutaneously inoculated into nude mice. After the tumor volume reached 100-150 mm3, the animals were randomly divided into several groups, and administered with the water soluble cabazitaxel derivatives of the following formula and cabazitaxel, respectively once on the same day (D0). The dosage and dosage regimen are shown in following table 6. The tumor volume was measured 2-3 times per week, the animals were weighed and the data were recorded until day 20 (D20) after grouping.
The tumor volume (V) was calculated according to the equation: V=1⁄2×a×b2, wherein a and b represent the length and the width, respectively.
T/C (%)=(T−T0)/(C−C0)×100, wherein T and C represent the tumor volume at the end of the experiment; and T0 and C0 represent the tumor volume at the beginning of the experiment.
The antitumor activity data are shown in following Table 6:











TABLE 6





The inhibitory activity of the water soluble cabazitaxel derivatives and cabazitaxel


on subcutaneously xenografted tumor of human prostate cancer PC-3 in nude mice.





























Time and












Compd.

route of












No.
Dosage
administration
n
R1
R2
R3
R4
R5
X
Y
C*
A





the control
—
D0 IV
—
—
—
—
—
—
—
—
—
—


group














cabazitaxel
 7 mg/kg
D0 IV
—
—
—
—
—
—
—
—
—
—


C7
13 mg/kg
D0 IV
2
Me
Me
t-BuO
Me
Me
H
F
R
MeSO3H


C8
13 mg/kg
D0 IV
2
Me
Me
t-BuO
Me
Me
H
F
S
MeSO3H


C9
13 mg/kg
D0 IV
2
Me
Me
t-BuO
Me
Me
H
F
R, S
MeSO3H


C32
13 mg/kg
D0 IV
2
Me
Me
t-BuO
Et
Me
H
F
R
maleic acid


C33
13 mg/kg
D0 IV
2
Me
Me
t-BuO
Et
Et
H
F
R
MeSO3H


C34
13 mg/kg
D0 IV
2
Me
Me
t-BuO
Me
i-Pro
H
F
R
HCl


C35
13 mg/kg
D0 IV
2
Me
Me
t-BuO
Me
Me
H
CF3
R
MeSO3H


C36
13 mg/kg
D0 IV
2
Me
Me
t-BuO
Me
Me
H
CHF2
R
p-toluene














sulfonic acid


C37
13 mg/kg
D0 IV
3
Me
Me
t-BuO
Me
Me
H
F
R
HCl


C38
13 mg/kg
D0 IV
4
Me
Me
t-BuO
Me
Me
H
F
R
sulfuric acid












Anti-tumor activity data


















Tumor





Tumor
Tumor

inhibitory
Weight



Compd.
volume
volume
T/C
percent
change



No.
(mm3) (D0)
(mm3) (D20)
(%) D20
(%)Δ D20
(%) D6






the control
111.3 ± 3.8
868.4 ± 58.7 
—
—
−17



group








cabazitaxel
111.7 ± 1.8
79.9 ± 24.8
−27
127
−13.8



C7
111.5 ± 2.2
31.5 ± 11.8
−67
154
−14.2



C8
111.9 ± 2.0
32.1 ± 12.1
−66
151
−14.4



C9
111.4 ± 2.3
32.5 ± 11.6
−66
151
−14.4



C32
112.4 ± 3.0
35.8 ± 12.7
−56
128
−14.7



C33
112.0 ± 2.4
38.6 ± 13.4
−50
114
−15.1



C34
111.7 ± 3.2
42.8 ± 11.9
−45
101
−15.4



C35
110.2 ± 4.6
42.3 ± 12.6
−46
105
−15.8



C36
116.7 ± 3.9
48.4 ± 14.7
−37
85
−16.1



C37
112.9 ± 4.4
55.3 ± 13.8
−28
64
−16.6



C38
111.9 ± 3.7
66.8 ± 12.4
−20
46
−16.8





D0: the time of administration for the first time;


ΔP(D20) = 0.000, compared to the control, using Student's t-test.


The numbers of mice at the beginning of the experiment: n = 10 in the control group, and n = 6 in the administered group.


C* is the configuration of the chiral carbon in the side chain of an amino acid.


Conclusion: the water soluble cabazitaxel derivatives have an inhibitory effect on human prostate cancer PC-3.






D0: the time of administration for the first time:
Δ: P(D20)=0.000, compared to the control using Student's t-test.
The numbers of mice at the beginning of the experiment: n=10 in the control group, and n=6 in the administered group.
C* is the configuration of the chiral carbon in the side chain of an amino acid.
Conclusion: the water soluble cabazitaxel derivatives have an inhibitory effect on human prostate cancer PC-3.
6.4. Test of the Inhibitory Activity of the Water Soluble Larotaxel and Tesetaxel Derivatives on Subcutaneously Xenografted Tumor of Human Non-Small Cell Lung Cancer NCI-H446 in Nude Mice
Purpose: to evaluate and compare the inhibitory activity of the water soluble larotaxel and tesetaxel derivatives on subcutaneously xenografted tumor of human non-small cell lung cancer NCI-H446 in nude mice.
Dosage Regimen and Experimental Procedures:
Human non-small cell lung cancer NCI-H446 cells were subcutaneously inoculated into nude mice. After the tumor volume reached 100-150 mm3, the animals were randomly divided into several groups, and administered with the water soluble larotaxel and tesetaxel derivatives of the following formulae, larotaxel, and tesetaxel, respectively once on the same day (D0). The dosage and dosage regimen are shown in the following table. The tumor volume was measured 2-3 times per week, the animals were weighed and the data were recorded until day 20 (D20) after grouping.
The tumor volume (V) was calculated according to the equation: V=1⁄2×a×b2, wherein a and b represent the length and the width, respectively.
T/C (%)=(T−T0)/(C−C0)×100, wherein T and C represent the tumor volume at the end of the experiment; and T0 and C0 represent the tumor volume at the beginning of the experiment.
The antitumor activity data are shown in following Table 7:











TABLE 7





The inhibitory activity of the water soluble larotaxel and tesetaxel derivatives, larotaxel, and tesetaxel


on subcutaneously xenografted tumor of human non-small cell lung cancer NCI-H446 in nude mice.





























Time and












Compd.

route of
Type of the











No.
Dosage
administration
parent ring
n
R2
R3
R4
R5
X
Y
C*
A





The control
—
D0 IV
—
—
—
—
—
—
—
—
—
—


group














larotaxel
 7 mg/kg
D0 IV
I-2
—
Ac
t-BuO
—
—
—
—
—
—


tesetaxel
 8 mg/kg
D0 IV
I-3
—
—
t-BuO
—
—
—
—
—
—


C39
12 mg/kg
D0 IV
I-2
2
Ac
t-BuO
Me
Me
H
F
R
HCl


C40
12 mg/kg
D0 IV
I-2
2
Ac
t-BuO
Et
Me
H
F
R, S
HCl


C41
12 mg/kg
D0 IV
I-2
2
Ac
t-BuO
Me
i-Pro
H
CF3
R, S
maleic acid


C42
12 mg/kg
D0 IV
I-2
3
Ac
t-BuO
Me
Me
H
F
S
phosphoric acid


C43
12 mg/kg
D0 IV
I-3
2
—
t-BuO
Me
Me
H
F
R
HCl


C44
12 mg/kg
D0 IV
I-3
2
—
t-BuO
Et
Et
H
F
S
HCl


C45
12 mg/kg
D0 IV
I-3
2
—
t-BuO
Me
i-Pro
H
CF3
R, S
succinic acid


















C46
12 mg/kg
D0 IV
I-3
3
—
t-BuO
—CH2CH2CH2CH2—
H
F
R, S
sulfuric acid












Anti-tumor activity data


















Tumor





Tumor
Tumor

inhibitory
Weight



Compd.
volume
volume
T/C
percent
change



No.
(mm3) (D0)
(mm3) (D20)
(%) D20
(%)Δ D20
(%) D5






The control
111.3 ± 3.8
868.4 ± 58.7 
—
—
−15.5



group








larotaxel
110.9 ± 2.4
33.3 ± 12.4
−64
153
−8.4



tesetaxel
112.8 ± 2.2
33.7 ± 12.1
−58
146
−7.4



C39
111.5 ± 2.2
34.9 ± 13.3
−62
148
−8.5



C40
112.4 ± 1.8
43.2 ± 12.7
−51
122
−9.1



C41
109.8 ± 2.9
51.7 ± 11.9
−34
67
−10.5



C42
109.8 ± 2.9
51.7 ± 9.8 
−22
53
−13.8



C43
111.5 ± 2.2
31.8 ± 11.1
−61
153
−8.2



C44
113.6 ± 2.8
41.3 ± 10.9
−49
123
−9.4



C45
112.3 ± 3.1
53.4 ± 12.4
−34
85
−11.4



C46
111.7 ± 2.8
61.2 ± 14.2
−23
58
−14.4





D0: the time of administration for the first time;


ΔP(D20) = 0.000, compared to the control, using Student's t-test.


The numbers of mice at the beginning of the experiment: n = 10 in the control group, and n = 6 in the administered group.


C* is the configuration of the chiral carbon in the side chain of an amino acid.


Conclusion: the water soluble larotaxel and tesetaxel derivatives have an inhibitory effect on human non-small cell lung cancer NCI-H446.






According to the results in the further experimentation, the inventors found that the taxane derivatives of the present invention have good water solubility, and can be dissociated in blood plasma to release a taxane parent drug, and thus are suitable as antitumor prodrugs.

## Claims

1. A water soluble taxane derivative of general formula (I-1), (I-2) or (I-3):



wherein,
R1 is H or methyl;
R2 is H, methyl or acetyl;
R3 is phenyl or OC(CH3)3;
X is H, C1-6 alkyl or F;
Y is F or C1-6 alkyl substituted with one or more F;
n is 1, 2, 3, 4, 5 or 6;
W is NR4R5.A or




R4 and R5 are each independently H, C1-6 alkyl optionally substituted with phenyl, or C3-6 cycloalkyl;
m is 0, 1, 2 or 3; and
A is a pharmaceutically acceptable acid.

2. The water soluble taxane derivative according to claim 1, characterized in that in general formula (I-1),
R1 is H, R2 is acetyl, and R3 is phenyl;
R1 is H, R2 is H, and R3 is OC(CH3)3; or
R1 is methyl, R2 is methyl, and R3 is OC(CH3)3.

3. The water soluble taxane derivative according to claim 1, characterized in that in general formula (I-2),
R2 is acetyl and R3 is OC(CH3)3.

4. The water soluble taxane derivative according to claim 1, characterized in that in general formula (I-3),
R3 is OC(CH3)3.

5. The water soluble taxane derivative according to claim 1, characterized in that the C1-6 alkyl is methyl, ethyl, propyl, isopropyl, butyl or isobutyl.

6. The water soluble taxane derivative according to claim 1, characterized in that the C3-6 cycloalkyl is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

7. The water soluble taxane derivative according to claim 1, characterized in that X is H, methyl or F.

8. The water soluble taxane derivative according to claim 1, characterized in that Y is F, CF3 or CHF2.

9. The water soluble taxane derivative according to claim 1, characterized in that R4 and R5 are each independently H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, benzyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

10. The water soluble taxane derivative according to claim 1, characterized in that:
X is H, methyl or F;
Y is F, CF3 or CHF2;
R4 and R5 are each independently H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, benzyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

11. The water soluble taxane derivative according to claim 1, characterized in that the pharmaceutically acceptable acid is hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, carbonic acid, acetic acid, propionic acid, methanesulfonic acid, lactic acid, benzensulfonic acid, p-toluene sulfonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid or malic acid.

12. The water soluble taxane derivative according to claim 1, characterized in that when X and Y are different, the carbon atom to which both X and Y are attached is in a single R configuration, in a single S configuration, or in both R and S configurations.

13. The water soluble taxane derivative according to claim 1, which is selected from the group consisting of:
2′-O-[4-N,N-dimethylamino-2(R)-fluorobutyryl]paclitaxel hydrochloride;
2′-O-[4-N,N-dimethylamino-2(R)-fluorobutyryl]paclitaxel methanesulfonate;
2′-O-[4-N,N-dimethylamino-2(R,S)-fluorobutyryl]paclitaxel hydrochloride;
2′-O-[4-N,N-dimethylamino-2(R,S)-difluoromethylbutyryl]paclitaxel citrate;
2′-O-[4-N,N-dimethylamino-2(R)-fluorobutyryl]docetaxel hydrochloride;
2′-O-[4-N,N-dimethylamino-2(R)-fluorobutyryl]docetaxel methanesulfonate;
2′-O-[4-amino-2(R,S)-trifluoromethylbutyryl]docetaxel methanesulfonate;
2′-O-[4-N,N-dimethylamino-2(R,S)-fluorobutyryl]cabazitaxel hydrochloride;
2′-O-[4-N,N-dimethylamino-2(R)-fluorobutyryl]larotaxel hydrochloride;
2′-O-[4-N,N-dimethylamino-2(R)-fluorobutyryl]tesetaxel hydrochloride;
2′-O-[4-N,N-dimethylamino-2(S)-fluorobutyryl]paclitaxel hydrochloride;
2′-O-[4-N,N-dimethylamino-2(S)-fluorobutyryl]docetaxel hydrochloride;
2′-O-[4-N,N-dimethylamino-2(R,S)-fluorobutyryl]docetaxel hydrochloride;
2′-O-[4-N,N-dimethylamino-2(R)-fluorobutyryl]cabazitaxel hydrochloride;
2′-O-[4-N,N-dimethylamino-2(S)-fluorobutyryl]cabazitaxel hydrochloride;
2′-O-[4-amino-2(R)-difluoromethylbutyryl]cabazitaxel sodium bisulfate salt;
2′-O-[4-N,N-diethylamino-2-methyl-2(R)-2-trifluoromethylbutyryl]cabazitaxel methanesulfonate;
2′-O-[4-N-methyl-N-ethylamino-2(R)-2-difluoroethylvaleryl]docetaxel hydrochloride;
2′-O-[4-N,N-dimethylamino-2(R)-fluorovaleryl]docetaxel methanesulfonate;
2′-O-[4-(aziridin-1-yl)amino-2,2-difluoroheptanoyl]docetaxel methanesulfonate;
2′-O-[4-N-methyl-N-cyclopentylamino-2-trifluoromethyl-2-fluorooctanoyl]paclitaxel methanesulfonate;
2′-O-[4-N,N-dimethylamino-2(R)-fluorovaleryl]larotaxel methanesulfonate;
2′-O-[4-N,N-dimethylamino-2(R)-fluorovaleryl]tesetaxel methanesulfonate;
2′-O-[4-N,N-dimethylamino-2(S)-fluorobutyryl]paclitaxel methanesulfonate;
2′-O-[4-N,N-dimethylamino-2(R,S)-fluorobutyryl]paclitaxel methanesulfonate;
2′-O-[4-N,N-dimethylamino-2(S)-fluorobutyryl]docetaxel methanesulfonate;
2′-O-[4-N,N-dimethylamino-2(R,S)-fluorobutyryl]docetaxel methanesulfonate;
2′-O-[4-N,N-dimethylamino-2(R)-fluorobutyryl]cabazitaxel methanesulfonate;
2′-O-[4-N,N-dimethylamino-2(S)-fluorobutyryl]cabazitaxel methanesulfonate;
2′-O-[4-N,N-dimethylamino-2(R,S)-fluorobutyryl]cabazitaxel methanesulfonate;
2′-O-[4-amino-2(R)-difluoromethylbutyryl]cabazitaxel sulfate;
2′-O-[4-N,N-diethylamino-2-methyl-2(R)-2-trifluoromethylbutyryl]cabazitaxel hydrochloride;
2′-O-[4-N-methyl-N-ethylamino-2(R)-2-difluoroethylvaleryl]docetaxel methanesulfonate;
2′-O-[4-N,N-dimethylamino-2(R)-fluorovaleryl]docetaxel hydrochloride;
2′-O-[4-(aziridin-1-yl)amino-2,2-difluoroheptanoyl]docetaxel hydrochloride;
2′-O-[4-N-methyl-N-cyclopentylamino-2-trifluoromethyl-2-fluorooctanoyl]paclitaxel hydrochloride;
2′-O-[4-N,N-dimethylamino-2(R)-fluorovaleryl]larotaxel hydrochloride;
2′-O-[4-N,N-dimethylamino-2(R)-fluorovaleryl]tesetaxel hydrochloride;
2′-O-[4-N-methyl-N-ethylamino-2(R)-fluorobutyryl]paclitaxel methanesulfonate;
2′-O-[4-N,N-diethylamino-2(S)-fluorobutyryl]paclitaxel fumarate;
2′-O-[4-N-methyl-N-isopropylamino-2(R,S)-fluorobutyryl]paclitaxel hydrochloride;
2′-O-[4-N,N-dimethylamino-2(R,S)-2-trifluoromethylbutyryl]paclitaxel p-toluene sulfonate;
2′-O-[4-N,N-dimethylamino-2(R,S)-2-difluoromethylbutyrvl]paclitaxel hydrochloride;
2′-O-[4-N,N-dimethylamino-2(R)-fluorovaleryl]paclitaxel maleate;
2′-O-[4-N,N-dimethylamino-2(S)-2-difluoromethylhexanoyl]paclitaxel sulfate;
2′-O-[4-N-methyl-N-ethylamino-2(R)-fluorobutyryl]docetaxel methanesulfonate;
2′-O-[4-N,N-diethylamino-2(S)-fluorobutyryl]docetaxel maleate;
2′-O-[4-N-methyl-N-isopropylamino-2(R)-fluorobutyryl]docetaxel hydrochloride;
2′-O-[4-N,N-dimethylamino-2(R)-2-trifluoromethylbutyryl]docetaxel methanesulfonate;
2′-O-[4-N,N-dimethylamino-2(R,S)-2-difluoromethylbutyryl]docetaxel hydrochloride;
2′-O-[4-N,N-dimethylamino-2(R,S)-fluorovaleryl]docetaxel methanesulfonate;
2′-O-[4-N,N-dimethylamino-2(S)-fluorohexanoyl]docetaxel sulfate;
2′-O-[4-N-methyl-N-ethylamino-2(R)-fluorobutyryl]cabazitaxel maleate;
2′-O-[4-N,N-diethylamino-2(R)-fluorobutyryl]cabazitaxel methanesulfonate;
2′-O-[4-N-methyl-N-isopropylamino-2(R)-fluorobutyryl]cabazitaxel hydrochloride;
2′-O-[4-N,N-dimethylamino-2(R)-2-trifluoromethylbutyryl]cabazitaxel methanesulfonate;
2′-O-[4-N,N-dimethylamino-2(R)-2-difluoromethylbutyryl]cabazitaxel p-toluene sulfonate;
2′-O-[4-N,N-dimethylamino-2(R)-fluorovaleryl]cabazitaxel hydrochloride;
2′-O-[4-N,N-dimethylamino-2(R)-fluorohexanoyl]cabazitaxel sulfate;
2′-O-[4-N-methyl-N-ethylamino-2(R,S)-fluorobutyryl]larotaxel hydrochloride;
2′-O-[4-N-methyl-N-isopropylamino-2(R,S)-2-trifluoromethylbutyryl]larotaxel maleate;
2′-O-[4-N,N-dimethylamino-2(S)-fluorovaleryl]larotaxel phosphate;
2′-O-[4-N,N-diethylamino-2(S)-fluorobutyryl]tesetaxel hydrochloride;
2′-O-[4-N-methyl-N-isopropylamino-2(R,S)-2-trifluoromethylbutyryl]tesetaxel succinate;
2′-O-[4-(pyrrolidin-1-yl)amino-2(R,S)-fluorovaleryl]tesetaxel sulfate;
2′-O-[4-N,N-diethylamino-2(R)-fluorobutyryl]paclitaxel hydrochloride;
2′-O-[3-N,N-dimethylamino-2(R,S)-fluoropropionyl]paclitaxel hydrochloride;
2′-O-[4-N-benzylamino-2(S)-fluorobutyryl]docetaxel hydrochloride;
2′-O-[3-(N,N-diethyl)amino-2(R,S)-trifluoromethylpropionyl]docetaxel hydrochloride;
2′-O-[4-(N-methyl)amino-2(R)-trifluoromethylbutyryl]paclitaxel hydrochloride;
2′-O-[3-(N-isopropyl)amino-2(R,S)-difluoromethylpropionyl]cabazitaxel hydrochloride;
2′-O-[5-(N,N-dimethyl)amino-2(R)-fluorovaleryl]paclitaxel hydrochloride;
2′-O-[4-(N,N-dimethyl)amino-2(S)-trifluoromethylbutyryl]cabazitaxel hydrochloride;
2′-O-[4-(N-isopropyl)amino-2(R,S)-trifluoromethylbutyryl]paclitaxel hydrochloride;
2′-O-[5-N,N-dimethylamino-2(S)-trifluoromethylvaleryl]docetaxel hydrochloride;
2′-O-[4-N-benzylamino-2-methyl-2(R,S)-fluorobutyryl]paclitaxel hydrochloride;
2′-O-[4-N,N-dimethylamino-2(R)-difluoromethylbutyryl]docetaxel hydrochloride;
2′-O-[3-cyclopentylamino-2-ethyl-2(R,S)-trifluoromethylpropionyl]docetaxel hydrochloride;
2′-O-[5-N-benzylamino-2-benzyl-2(R)-difluoromethylvaleryl]cabazitaxel hydrochloride;
2′-O-[4-(4-piperidin-1-yl)-2(S)-trifluoromethylbutyryl]cabazitaxel hydrochloride; and
2′-O-[4-N,N-dimethylamino-2(S)-fluorobutyryl]larotaxel hydrochloride; and
2′-O-[4-N,N-dimethylamino-2(R)-fluorobutyryl]tesetaxel hydrochloride.

14. A method for treating tumor, comprising administering to a patient the water soluble taxane derivative according to claim 1.