Patent Application
20220378735
The present invention relates to a cabazitaxel weakly-alkaline derivative and a preparation thereof, in particular relates to synthesis of the cabazitaxel weakly-alkaline derivative, a liposome preparation including the derivative and use of the cabazitaxel weakly-alkaline derivative in a drug delivery system. The present invention belongs to the field of medical technology.
Cabazitaxel (CTX) is a new generation of tubulin-binding agent of taxanes. Compared with paclitaxel and docetaxel, due to methylation of 7- and 10-hydroxyl groups, CTX has shown a low affinity for P-glycoprotein which closely associates with drug resistance in tumors, reduced efflux of drugs, and thus can better overcome the defect of multi-drug resistance. In June 2010, the U.S. Food and Drug Administration (FDA) has approved the use of cabazitaxel injections (trade name: Jevtana®) produced by Sanofi-Aventis, France in combination with prednisone for treating hormone-resistant prostate cancer. However, Tween 80 in the preparation causes toxic and side effects associated with excipients, which severely limits the clinical application of the drug. In order to overcome the shortcomings of the cabazitaxel injection, it is necessary to redesign its core molecule and select a suitable preparation to increase the clinical application potential.
Liposomes have a phospholipid bilayer structure similar to that of a biological cell membrane and may wrap drugs to isolate them from surrounding tissue and reduce the irritation of the drugs by controlling the release of drugs. Compared with other preparations, a liposome preparation has good biocompatibility. By PEG-modifying the phospholipid, the liposome may be endowed with a long-circulation function, which can significantly prolong a circulation time of the drug in the body, thereby making the nano-sized liposome pass through a tumor blood vessel to reach a tumor site by means of enhanced permeability and retention effect (EPR effect) of tumors. Therefore, the tumors' targeting property is indirectly improved, the drug's uptake in the tumor site is increased, and the drug's release in normal tissues is reduced, thereby achieving the effects of “reducing toxicity and increasing efficacy”.
At present, for most of liposome preparations used clinically, drugs are encapsulated through an active drug loading method. Compared with a passive drug loading method, the active drug loading method has the characteristics of high drug loading capacity, high encapsulation rate, and good stability of the preparation. In addition, as the drug is encapsulated in an internal aqueous phase of the liposome, the drug is prevented from leaking during a systemic circulation process. In the active drug loading method, it usually requires a more stable combination of the drug and a trapping agent in the internal aqueous phase, so that drug molecules present in the external aqueous phase enter the internal aqueous phase of the liposome through transmembrane driving force under certain conditions and stably exist. Cabazitaxel, as an electrically neutral compound, does not have weak acidity or weak basicity, thus it needs to be weakly alkalized so that it can be stably encapsulated in the internal aqueous phase of the liposome through the active drug loading method. Through the EPR effect, the nano-sized liposome preparation is targeted to the tumor site and then releases the encapsulated cabazitaxel derivative, which may release the active drug again after being hydrolyzed by esterase in vivo.
In order to overcome the shortcomings of existing cabazitaxel injections, exemplary embodiments provided according to the present invention provide a cabazitaxel weakly-alkaline derivative and prepare the derivative into a liposome of the derivative by encapsulating it, so as to obtain a nanoliposome preparation having a high drug loading capacity, a high encapsulation rate and good stability.
Exemplary embodiments provided according to the present invention weak-base modify cabazitaxel to prepare it into a nano-drug delivery system of liposomes and endow it with long-circulation properties in blood for anti-tumor research.
In some embodiments disclosed herein, a cabazitaxel weakly-alkaline derivative or a pharmaceutically acceptable salt thereof is provided. The cabazitaxel weakly-alkaline derivative has a structural formula as follows:
The linking group is C1-C4 alkyl, C3-C6 cycloalkyl or phenyl; and the [N] is N-methylpiperazinyl, piperidinyl, 4-(1-piperidinyl) piperidinyl, morpholinyl, tetrahydropyrrolyl, or other tertiary amine structure.
In some embodiments disclosed herein, a method for synthesizing a cabazitaxel weakly-alkaline derivative is provided. The cabazitaxel weakly-alkaline derivative has a structural formula as follows:
The method includes: performing esterification reaction between 4-(4-methylpiperazinylmethyl) benzoyl chloride and cabazitaxel under the catalysis of DMAP or triethylamine; and then performing separation and purification to obtain the cabazitaxel weakly-alkaline derivative. The whole reaction process is performed under the protection of N2.
In some embodiments disclosed herein, a liposome of a cabazitaxel weakly-alkaline derivative is provided. The cabazitaxel weakly-alkaline derivative is prepared into a liposome and the cabazitaxel weakly-alkaline derivative has a structural formula as follows:
The liposome includes the cabazitaxel weakly-alkaline derivative, phospholipid, cholesterol, and PEGylated phospholipid. The liposome is prepared by the steps of: (1) preparing a blank liposome having a gradient; (2) preparing an ethanol solution of the cabazitaxel weakly-alkaline derivative; and (3) incubating the ethanol solution of the cabazitaxel weakly-alkaline derivative and the blank liposome having a gradient.
In some embodiments disclosed herein, a use of the previously described cabazitaxel weakly-alkaline derivative, pharmaceutically acceptable salt thereof, or liposome of the cabazitaxel weakly-alkaline derivative is provided in the preparation of a drug delivery system.
In some embodiments disclosed herein, a use of the previously described cabazitaxel weakly-alkaline derivative, pharmaceutically acceptable salt thereof, or liposome of the cabazitaxel weakly-alkaline derivative is provided in preparation of anti-tumor drugs.
The above-mentioned and other features and advantages of this invention, and the manner of attaining them, will become more apparent and the invention will be better understood by reference to the following description of embodiments of the invention taken in conjunction with the accompanying drawings, wherein:
Corresponding reference characters indicate corresponding parts throughout the several views. The exemplifications set out herein illustrate embodiments of the invention and such exemplifications are not to be construed as limiting the scope of the invention in any manner.
For a cabazitaxel weakly-alkaline derivative provided according to the present invention, cabazitaxel is connected to a weakly-alkaline intermediate through an ester bond, and the ester bond may be broken under the action of esterase in vivo so as to release an active drug. A structural formula of the cabazitaxel weakly-alkaline derivative is as follows:
wherein
Further, according to the present invention, a cabazitaxel weakly-alkaline derivative having the following structure is provided:
A synthesis method of the cabazitaxel weakly-alkaline derivative according to the present invention is as follows:
under the catalysis of DMAP, performing esterification reaction between 4-(4-methylpiperazinylmethyl) benzoyl chloride and cabazitaxel, and then performing separation and purification to obtain the cabazitaxel weakly-alkaline derivative, wherein the whole reaction process is performed under the protection of N2, and DMAP may be replaced with triethylamine.
Further, the present invention provides a liposome including a cabazitaxel weakly-alkaline derivative, wherein the liposome includes the cabazitaxel weakly-alkaline derivative, phospholipid, cholesterol, PEGylated phospholipid, and the like. The weight ratio of the derivative to total lipids is 1:4-12, and the total lipids is a sum of phospholipid, cholesterol, and PEGylated phospholipid. The phospholipid, cholesterol and PEGylated phospholipid are used in conventional amounts in the art.
The present invention further provides a preparation method of a liposome of the cabazitaxel weakly-alkaline derivative, comprising:
In some embodiments, in step (2), the internal aqueous phase solution may be a citric acid solution, an ammonium sulfate solution, a sulfobutyl ether-β-cyclodextrin triethylammonium salt solution, or a sucrose octasulfate triethylammonium salt solution.
In some embodiments, the internal aqueous phase solution is an ammonium sulfate solution.
In sonic embodiments, in step (3), the external aqueous phase solution may be a sucrose solution, HEPES buffer, phosphate buffer, or acetate buffer.
In some embodiments, the external aqueous phase solution is a sucrose solution.
In some embodiments, in step (4), a solvent of the organic solution may be methanol, ethanol, acetone, tetrahydrofuran, acetonitrile, or DMSO.
In some embodiments, a solvent of the organic solution is ethanol.
In the present invention, cabazitaxel is prepared into the weakly-alkaline derivative for preparing a liposome, which can not only avoid the side effects caused by Tween 80 in a cabazitaxel injection, but also increase the maximum tolerated dose of a drug. Thus, the anti-tumor effect of the drug is enhanced and great clinical application potential is achieved.
The liposome nano-drug delivery system provided according to the present invention has the advantages as follows: (1) the particle size is small and uniform (less than 100 nm), which is beneficial for enrichment of the drug to a tumor site through the EPR effect; (2) the drug loading capacity is high, which is beneficial for reduction of adverse reactions caused by excipients and biological materials; (3) complete drug encapsulation can be acquired, and good stability and ease industrialization are achieved; (4) the uptake by a reticuloendothelial system is effectively avoided, a long circulation effect is achieved in blood and the probability that the drug reaches the tumor site is increased; and (5) compared with commercially available preparations, the liposome nano-drug delivery system provided according to the present invention has an improved anti-tumor effect and reduced toxic and side effects.
The following embodiments are intended to further illustrate the present invention without limiting the present invention in any way.
Cabazitaxel (200 mg, 0.24 mmol) and 4-(4-methylpiperazinylmethyl) benzoyl chloride (156 mg, 0.48 mmol) were weighed and dissolved in dichloromethane; 0.25 mL of triethylamine was added thereto; under an ice bath, a dichloromethane solution of DMAP (5.9 mg, 0.048 mmol) was slowly added dropwise to the thus obtained mixture; and stirring was performed at the room temperature overnight under N2 protection. After the reaction was completed, separation and purification were performed through column chromatography to acquire a cabazitaxel derivative in the form of a white powder (yield: 95.01%). The structure of the compound in Example 1 was determined through a nuclear magnetic resonance hydrogen spectrum. The results are shown in
1H NMR (Chloroform-d,400 MHz) δ8.11 (2H,d,J=7.4 Hz), 7.94 (2H,d,J=8.1 Hz), 7.61 (1H,t,J=7.4 Hz), 7.51 (1H,d,J=7.8 Hz), 7.48 (1H,d,J=7.8 Hz), 7.45-7.35(6H,m), 7.32-7.27(1H,m), 6.26 (1H,t,J=9.1 Hz), 5.64 (1H,d,J=7.0 Hz), 5.50 (1H,d,J=3.4 Hz), 5.43(1H,d,J=9.4 Hz), 5.30 (1H,s), 5.00(1H,d,J=8.8 Hz), 4.83 (1H,s), 4.31 (1H,d,J=8.4 Hz), 4.17(1H,d,J=8.3 Hz), 3.97-3.88(1H,m), 3.86 (1H,d,J=7.0 Hz), 3.56(2H,s), 3.43(3H,s), 3.31(3H,s), 2.82-2.62(2H,m), 2.54-2.38(6H,m), 2.31(3H,s), 2.25(1H,m), 2.04(3H,s), 1.82(1H,m), 1.79(1H,m), 1.71 (3H,s), 1.64 (2H,s), 1.36 (9H,s), 1.26(3H,s), 1.21(6H,s).
Cabazitaxel (200 mg, 0.24 mmol) and 4-piperidylpiperidinecarbonyl chloride (111 mg, 0.48 mmol) were weighed and dissolved in dichloromethane; 0.25 mL of triethylamine was added thereto; under an ice bath, a dichloromethane solution of DMAP (5.9 mg, 0.048 mmol) was slowly added dropwise to the thus obtained mixture; and stirring was performed at the room temperature overnight under N2 protection. After the reaction was completed, separation and purification were performed through column chromatography to acquire a cabazitaxel derivative in the form of a white powder (yield: 95%). The structure of the compound in Example 2 was determined through a nuclear magnetic resonance hydrogen spectrum. The results are shown in
1H NMR (400 MHz, Chloroform-d) δ8.07-8.00 (m,2H), 7.54(t,J=7.4 Hz, 1H), 7.43 (t,J=7.6 Hz,2H), 7.33 (t,J=7.6 Hz,2H), 7.22 (m,3H), 6.32-5.95 (m, 1H), 5.56 (d,J=7.0 Hz,1H), 5.44-5.26 (m,2H), 5.23 (s,1H), 5.20 (d,J=3.8 Hz, 1H), 4.92 (d,J=9.4 Hz,1H), 4.74(s,1H), 4.23 (d,J=8.4 Hz,1H), 4.09 (d,J=8.4 Hz,2H), 3.83 (dd,J=10.7,6.4 Hz,1H), 3.76 (d,J=7.0 Hz,1H), 3.36(s,3H), 3.22(s, 3H), 2.77-2.49 (m,4H), 2.36 (s,3H), 2.27-2.05 (m,1H), 1.92(s,3H), 1.77-1.66 (m,1H), 1.64(s,3H), 1.55(s,1H), 1.52-1.39 (m,2H), 1.28(s,9H), 1.19(s,3H), 1.14 (s,3H), 1.12(s,3H).
Cabazitaxel (200 mg, 0.24 mmol) and 4-methylpiperazine-1-carbonyl chloridel (78 mg, 0.48 mmol) were weighed and dissolved in dichloromethane; 0.25 mL of triethylamine was added thereto; under an ice bath, a dichloromethane solution of DMAP (5.9 mg, 0.048 mmol) was slowly added drop wise to the thus obtained mixture; and stifling was performed at the room temperature overnight under N2 protection. After the reaction was completed, separation and purification were performed through column chromatography to acquire a cabazitaxel derivative in the form of a white powder (yield: 93.8%). The structure of the compound in Example 3 was determined through a nuclear magnetic resonance hydrogen spectrum. The results are shown in
1H NMR (400 MHz,Chloroform-d) δ8.03 (d,J=7.5 Hz,2H), 7.54 (t,J=7.4 Hz, 1H), 7.43 (t,J=7.6 Hz,2H), 7.32 (t,J=7.6 Hz,2H), 7.26-7.20 (m,3H), 6.17(t,J=9.3 Hz,1H), 5.56 (d,J=7.0 Hz,1H), 5.36 (s,1H), 5.25(s,1H), 5.23(s,1H), 4.92(dd, J=9.5,2.0 Hz,1H), 4.74(s,1H), 4.23 (d,J=8.4 Hz,1H), 4.09 (d,J=8.4 Hz,1H), 3.83 (dd,J=10.7,6.3 Hz,1H), 3.76(d,J=7.0 Hz,1H), 3.39 (s,2H), 3.36 (s,3H), 3.22 (s,3H), 2.62 (ddd,J=14.1,9.8,6.3 Hz,1H), 2.36(m,1H), 2.35(s,3H), 2.22(s, 3H), 2.15 (t,J=7.6 Hz,2H), 2.00 (m,1H), 1.92 (s,3H), 1.78-1.66 (m,2H), 1.64(s, 3H), 1.52(s,1H), 1.28 (s,9H), 1.19(s,3H), 1.14(s,3H), 1.12(s,3H).
The preparation method of the liposomes of cabazitaxel derivate derivative according to the present example included the following steps:
The liposomes preparation prepared in Example 4 was subjected to sterile filtration and stored at 4° C. for 60 days. During this period, a change in particle size was measured through a dynamic light scattering method, and a change in encapsulation rate was measured through high-performance liquid chromatography. The results are shown in
10 healthy male rats with a weight of 200-250 g for each one, were randomly divided into 2 groups with 5 rats in each group, and these two groups of rats were injected with a commercially available preparation and the CN1-liposomes prepared in Example 4 at tail veins respectively, wherein the equivalent dosage of cabazitaxel was 5 mg/kg. Blood was taken from the orbits at a prescribed time and centrifuged to acquire plasma, and the concentration of a drug in the plasma was measured through high-performance liquid chromatography-mass spectrometry.
The results are shown in Table 1. A circulation time of the liposomes preparation in the body was significantly prolonged, and the area under the concentration-time curve (AUC) of the liposome preparation was significantly improved. The experimental results showed that the liposome preparation can significantly prolong the circulation time of the drug in the blood and increases the possibility that the drug accumulates to the tumor site through the EPR effect.
Mouse prostate cancer cells (RM-1, 5*106 cells/100 μL PBS) were inoculated subcutaneously on the right ventral sides of male C57BL/6 mice. After the tumors grew to 60-80 mm3, the mice were randomly divided into 6 groups with 5 mice in each group: a blank control group, a CN1-solution group, a commercially available preparation group, a CN1-liposomes group, a CN2-liposomes group, and a CN3-liposomes group. The drugs were administered every three days for a total of 5 times, wherein the equivalent dosage of cabazitaxel was 6 mg/kg, and the high dosage was three times of the low dosage. After administration, the states of the mice were observed, the mice were weighed and the volumes of the tumors were measured. At the end of the last dosing cycle, the mice were killed, tumors and major organs were stripped for analysis and evaluation.
The results are shown in
As shown in
While this invention has been described with respect to at least one embodiment, the present invention can be further modified within the spirit and scope of this disclosure. This application is therefore intended to cover any variations, uses, or adaptations of the invention using its general principles. Further, this application is intended to cover such departures from the present disclosure as come within known or customary practice in the art to which this invention pertains and which fall within the limits of the appended claims.
| Number | Date | Country | Kind |
|---|---|---|---|
| 201911218994.3 | Dec 2019 | CN | national |
This is a continuation of International Patent Application Publication No. PCT/CN2020/132491 entitled “WEAK ALKALINE CABAZITAXEL DERIVATIVE AND FORMULATION THEREOF” filed on Nov. 27, 2020, which is incorporated in its entirety herein by reference. International Patent Application Publication No. PCT/CN2020/132491 claims priority to Chinese Patent Application No. CN 201911218994.3 filed on Dec. 3, 2019, which is incorporated in its entirety herein by reference.
| Number | Date | Country | |
|---|---|---|---|
| Parent | PCT/CN2020/132491 | Nov 2020 | US |
| Child | 17831962 | US |
