Paclitaxel derivative as well as preparation method and application thereof

Abstract

The invention relates to the field of medicines, and particularly discloses a paclitaxel derivative shown in a formula (I) as well as a preparation method and application of the paclitaxel derivative. A baccatin diazo compound, imine and alcohol are used as raw materials, transition metal rhodium is used as a catalyst, and the paclitaxel derivative is obtained after a period of time of reaction at a certain temperature. According to the invention, efficient modular synthesis of the paclitaxel derivative can be realized. Benefited from the rapid synthesis method, large-scale anticancer activity screening is carried out on the paclitaxel derivatives, the antitumor activity of the paclitaxel derivatives in a cell level and a mouse model is superior to that of paclitaxel, and part of the paclitaxel derivatives show better anticancer activity in a paclitaxel drug-resistant cell model. The discovery of novel taxane medicines is facilitated.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to a paclitaxel derivative having the formula (I), its preparation method, and its application. Background Technology

[0002] Cancer has become the leading cause of death threatening human health in the 21st century. According to data released by the International Agency for Research on Cancer (IARC) of the World Health Organization, there were 19.29 million new cancer cases and 9.96 million cancer deaths globally in 2020. In China alone, there were 4.57 million new cancer cases and approximately 3 million cancer deaths in 2020, accounting for 23.7% and 30.2% of the global total, respectively. Currently, there are nearly 100 drugs used to treat cancer, and various innovative therapies are constantly emerging. In fact, chemotherapy remains one of the most important systemic anti-cancer therapies, with paclitaxel being a leading example, hailed as "the most effective anti-cancer drug discovered by mankind to date," and the world's best-selling herbal anti-cancer drug, with annual global sales exceeding 30 billion yuan. In addition to paclitaxel, several paclitaxel derivatives have been developed and launched, such as docetaxel launched in 1996 and cabazitaxel launched in 2010, while several other derivatives are in clinical research stages.

[0003] Paclitaxel was initially extracted from the bark of the Pacific yew tree by American chemists Wall and Vani. In 1971, Wall and Vani determined its structure using X-ray diffraction and nuclear magnetic resonance (NMR). In 1992, Holton's team developed a semi-synthetic route for paclitaxel, which was subsequently approved by the FDA for the treatment of ovarian cancer the following year. In 1994, Nicolaou's team achieved the first total synthesis of paclitaxel (39 steps). However, the total synthetic route for paclitaxel suffers from high cost and long processing time, making it difficult to use for industrial production. Compared to total synthesis, the semi-synthetic route for paclitaxel is shorter and more economically feasible. Currently, some paclitaxel on the market is supplied through semi-synthesis. The challenge in semi-synthesis research lies in how to efficiently construct two consecutive chiral centers in the side chain. To date, various side-chain precursors have been developed: phenylisoserine methyl ester type (linear), β-lactam type (tetracyclic), oxazoline type (pentacyclic), or oxazolidinyl type (pentacyclic). Currently, the synthesis of chiral side chains faces the following challenges: long synthetic routes and low overall yields; expensive chiral catalysts are unsuitable for large-scale synthesis; while chiral prosthetic routes can achieve process scale-up, they lack atom economy; and linear side-chain precursors have low coupling efficiency with the parent nucleus. Although the semi-synthetic process of paclitaxel is mature, synthesizing paclitaxel derivatives using traditional routes requires the synthesis of different chiral side chains, thus greatly limiting the variety of paclitaxel derivatives. Therefore, developing new semi-synthetic methods for paclitaxel and efficiently synthesizing paclitaxel and its derivatives is of great significance for the discovery of novel taxane drugs. Our team has a long-standing research foundation in the semi-synthesis of paclitaxel and last year filed a patent application for a one-step synthesis technique for paclitaxel (patent publication number: CN114573631A).

[0004] This technology still has the following shortcomings: 1. Poor chemoselectivity and stereoselectivity, and generally low reaction yield (crude yield of about 60%, including four diastereomers of paclitaxel derivatives), which greatly complicates purification and separation, resulting in low synthesis efficiency and limited practicality. 2. Limited substrate applicability, making it difficult to achieve modular and efficient synthesis of paclitaxel derivatives. Summary of the Invention

[0005] In order to overcome the shortcomings of the prior art, the primary objective of this invention is to provide a paclitaxel derivative.

[0006] A second objective of this invention is to provide a method for preparing the aforementioned paclitaxel derivatives.

[0007] A third objective of this invention is to provide the application of the aforementioned paclitaxel derivatives in the preparation of antitumor drugs. Some of the paclitaxel derivatives provided by this invention exhibit significantly superior antitumor activity compared to paclitaxel at the cellular level and in mouse models; therefore, the paclitaxel derivatives provided by this invention have great potential for application in antitumor activity.

[0008] This invention overcomes the following technical challenges in the synthesis of paclitaxel derivatives: 1. Finding a co-catalyst that is compatible with the reaction system and possesses high reactivity and good stereoselectivity. Ensuring both high reactivity and high stereoselectivity is a highly challenging problem in stereochemical synthesis. 2. Finding a solvent compatible with the reaction system. This reaction involves a highly reactive carbene intermediate, which is highly likely to react with the solvent, generating byproducts and reducing reaction efficiency. Furthermore, the poor solubility of the diazo compounds used in this technology complicates the selection of solvents.

[0009] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0010] This invention provides a paclitaxel derivative having the structure shown in formula (I), which is as follows:

[0011]

[0012] in,

[0013] R1 group is hydrogen, substituted silicon group, C1-C4 alkyl, -CH2-C3-C5 cycloalkyl, or C3-C8 cycloalkyl;

[0014] The R2 group can be hydrogen, C1-C4 alkyl, C3-C8 cycloalkyl, C1-C4 acyl, or C3-C8 cyclocarboxyl.

[0015] The R3 group can be hydrogen, substituted silicon, C1-C4 alkyl, C3-C8 cycloalkyl, C1-C4 acyl, or C3-C8 cyclocarboxyl.

[0016] The R4 group is phenyl, substituted phenyl, pyridine, pyrimidine, pyrrole, pyrazole, pyridazine, furan, substituted furan, thiophene, substituted thiophene, C6-C12 fused ring, C1-C4 alkoxy, wherein the substituent is independently selected from -F, -Cl, and -Br;

[0017] Ar1 group is phenyl, substituted phenyl, pyridine, pyrimidine, pyrrole, pyrazole, pyridazine, furan, substituted furan, thiophene, substituted thiophene, substituted indole, C6-C12 fused ring, wherein the substituent is independently selected from -F, -Cl, -Br, -CH3, -OCH3.

[0018] Preferably, the R1 group is hydrogen, methyl, ethyl, propyl, isopropyl, cyclopropyl, -CH2-cyclobutyl, tert-butyl, trimethylsilyl, triethylsilyl, tert-butyldimethylsilyl, or triphenylsilyl.

[0019] The R2 group is hydrogen, methyl, ethyl, isopropyl, cyclopropyl, formyl, acetyl, cyclopropylformyl, cyclobutylformyl, or cyclopentanoyl.

[0020] The R3 group is hydrogen, triethylsilyl, methyl, ethyl, isopropyl, cyclopropyl, formyl, acetyl, cyclopropylformyl, cyclobutylformyl, or cyclopentanoyl.

[0021] The R4 group is phenyl, substituted phenyl, 2-pyridine, 3-pyridine, 4-pyridine, pyrimidine, pyrrole, pyrazole, pyridazine, 2-furan, 2-substituted furan, 3-furan, 3-substituted furan, 2-thiophene, 2-substituted thiophene, 3-thiophene, 3-substituted thiophene, C6-C12 fused ring, tert-butoxy, wherein the substituent is independently selected from -F, -Cl, and -Br;

[0022] The Ar1 group is phenyl, substituted phenyl, 2-pyridine, 3-pyridine, 4-pyridine, pyrimidine, pyrrole, pyrazole, pyridazine, 2-furan, 2-substituted furan, 3-furan, 3-substituted furan, 2-thiophene, 2-substituted thiophene, 3-thiophene, 3-substituted thiophene, substituted indole, C6-C12 fused ring, wherein the substituent is independently selected from -F, -Cl, -Br, -CH3, -OCH3.

[0023] More preferably, the taxane derivative having the formula (I) is selected from compounds with the following structures:

[0024]

[0025] The present invention also provides a method for preparing paclitaxel derivatives having the formula (I), the method comprising the following steps: preparing a mixed solution a by dissolving compounds of formulas 1, 2 and 3 in an organic solvent according to the following reaction formulas; and adding a metal catalyst, an additive and... Molecular sieve Mixture b is obtained by dispersing the mixture in the organic solvent; mixture a is added to mixture b at -20℃ to 40℃ to obtain a paclitaxel derivative with a protecting group, and finally, the paclitaxel derivative of formula (I) is obtained under the action of pyridine hydrogen fluoride. The reaction route is shown below:

[0026]

[0027] The definitions of R1, R2, R3, R4, and Ar1 are as described above.

[0028] In this invention, compound 1 is a bacasine diazo compound, compound 3 is an imine, and compound 2 is an alcohol.

[0029] Wherein, the reaction molar ratio of the compound shown in Formula 1, the compound shown in Formula 2, the compound shown in Formula 3, the metal catalyst, and the additive is (0.5-2):(1-6):(1-4):(0.05-0.3):(0.1-0.5); preferably, the reaction molar ratio of the compound shown in Formula 1, the compound shown in Formula 2, the compound shown in Formula 3, the metal catalyst, and the additive is 1:6:4:0.05:0.1.

[0030] The metal catalyst includes one or more of Rh2(OAc)4, Rh2(oct)4, Rh2(esp)2, Rh2(Piv)4, Cu(OTf)2, Pd(ally)2Cl2, Cu(MeCN)4BF4, etc.; preferably, the metal catalyst is Rh2(OAc)4.

[0031] The additive includes one or more of AgPF6, AgBF4, AgOTf, AgTFA, AgClO4, AgSbF6, AgOTf, Cu(OTf)2, Mg(OTf)2, Sc(OTf)3, Fe(OTf)2, Yb(OTf)2, La(OTf)2, Bi(OTf)3, In(OTf)2, LiOTf, Zn(OTf)2, Hg(OTf)2, Ni(OTf)2, NaOTf, and Ca(OTf)2; preferably, the additive is Ni(OTf)2.

[0032] The organic solvent includes one or more of dichloromethane, trichloromethane, ethyl acetate, tetrahydrofuran, toluene, xylene, chlorobenzene, fluorobenzene, trifluorotoluene, and 1,4-dioxane; preferably, the organic solvent is trifluorotoluene.

[0033] The reaction temperature is -20℃ to 40℃; preferably, the reaction temperature is 25℃.

[0034] The reaction time is 0.5-4 hours, until the compound shown in Formula 1 is completely consumed.

[0035] The present invention also provides paclitaxel derivatives prepared by the above method.

[0036] The present invention also provides a pharmaceutical / pharmaceutical composition comprising the above-described paclitaxel derivative having formula (I) and a pharmaceutically acceptable carrier.

[0037] Furthermore, the drug or pharmaceutical composition may also contain physiologically compatible excipients, including buffers, diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption enhancers, surfactants, adsorbents, lubricants, solubilizers, stabilizers, and other pharmaceutical matrices.

[0038] The aforementioned carriers are acceptable functional pharmaceutical excipients in the pharmaceutical field, including surfactants, suspending agents, emulsifiers, and some novel pharmaceutical polymers, such as cyclodextrin, chitosan, polylactic acid (PLA), polyglycolic acid-polylactic acid copolymer (PLGA), hyaluronic acid, etc.

[0039] The formulations of the drug / drug composition include capsules, tablets, aqueous suspensions, solutions, rectal suppositories, enemas, ointments, lotions, creams, nasal sprays, and inhalers. Preferably, they are solutions and ointments.

[0040] Furthermore, the drug or drug composition may be introduced into the body, such as into muscles, intradermal tissues, subcutaneous tissues, veins, or mucous membranes, by means of injection, spray, nasal drops, eye drops, penetration, absorption, or physical or chemical mediated methods; or it may be introduced into the body after being mixed with or encapsulated by other substances.

[0041] Furthermore, the drug or drug composition may be used alone or in combination with other drugs such as doxorubicin, cyclophosphamide, carboplatin, cisplatin, capecitabine, fluorouracil, or gemcitabine.

[0042] The present invention also provides the use of the above-described paclitaxel derivatives having formula (I) or drug / drug composition in the preparation of antitumor drugs.

[0043] Specifically, the tumor includes, but is not limited to, triple-negative breast cancer, ovarian cancer, lung cancer, colorectal cancer, melanoma, head and neck cancer, lymphoma, and brain tumor. Preferably, it is triple-negative breast cancer.

[0044] This invention also provides the use of the above-mentioned paclitaxel derivatives, or the above-mentioned preparation method, or the drug / drug composition in the preparation of a drug for inhibiting tumor cell proliferation. Preferably, it is for triple-negative breast cancer.

[0045] Preferably, the tumor cells include, but are not limited to, triple-negative breast cancer, ovarian cancer, lung cancer, colorectal cancer, melanoma, head and neck cancer, lymphoma, and brain tumor.

[0046] Preferably, in the above applications, the taxane derivative having the formula (I) is selected from compounds with the following structures:

[0047]

[0048] Experiments have demonstrated that the paclitaxel derivatives of formula (I) of this invention have a good inhibitory effect on a variety of human cancer cells, with an IC50 value of [missing information]. 50 The value can be as low as 15.18 nM, which can be used to prepare anti-breast cancer drugs for application.

[0049] This invention, through experiments using a Balb / c mouse subcutaneous cell transplantation tumor model, discovered that a class of paclitaxel derivatives with formula (I) can effectively inhibit the growth of breast cancer cells.

[0050] Compared with the prior art, the beneficial effects of the present invention are:

[0051] This invention proposes a class of taxane derivatives having the formula (I) and a highly efficient synthetic method for these taxane derivatives. The bioactivity of the taxane derivatives was evaluated at the cellular level and in mouse models, and it was found that some derivatives have better antitumor activity than taxanes, which is helpful for the discovery of novel taxane drugs.

[0052] This invention, through further optimization and screening, develops a new generation of paclitaxel semi-synthetic method. This method improves the following technical parameters: 1. It proposes a more efficient co-catalyst, significantly increasing the reaction yield (20-40%); 2. It proposes a reaction solvent that is more favorable to stereoselectivity. In existing technology (CN114573631B), the ratio of the four diastereomers of the paclitaxel intermediate is approximately 25:25:25:25. After solvent optimization, this invention can improve this ratio to approximately 83:12:4:1, with the major isomer having the same stereoconfiguration as paclitaxel. These improvements in technical parameters reduce the difficulty of paclitaxel separation and greatly improve synthetic efficiency. Furthermore, the innovative method proposed in this invention has a wide substrate applicability and can be applied to the rapid modular synthesis of paclitaxel derivatives. Its application in the screening of anticancer activity of paclitaxel derivatives has revealed derivatives with better anticancer activity than paclitaxel. Attached Figure Description

[0053] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0054] Figure 1 The antiproliferative effect of paclitaxel and its derivatives on tumor cells in an in vitro model;

[0055] Figure 2 This describes tumor changes during administration in a mouse subcutaneous breast cancer cell xenograft model. Detailed Implementation

[0056] The specific embodiments of the present invention will be further described below. It should be noted that these descriptions are for the purpose of aiding understanding the present invention, but do not constitute a limitation thereof. Furthermore, the technical features involved in the various embodiments of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0057] The following description of at least one exemplary embodiment is merely illustrative and is in no way intended to limit the invention or its application or use. All other embodiments obtained by those skilled in the art based on the embodiments of this invention without inventive effort are within the scope of protection of this invention.

[0058] Unless otherwise specified, the experimental methods used in the following embodiments are conventional methods, and the experimental materials used in the following embodiments are all available through conventional commercial channels.

[0059] This invention relates to the pharmaceutical field, specifically proposing a paclitaxel derivative, its preparation method, and its application. The derivative is shown in general formula (I). Using bacasine diazo compound, imine, and alcohol as raw materials, and rhodium as a catalyst, the paclitaxel derivative is obtained after reacting at a certain temperature for a period of time. This invention enables the efficient and modular synthesis of paclitaxel derivatives. Thanks to this rapid synthesis method, this invention allows for large-scale screening of the anticancer activity of paclitaxel derivatives, and proposes that the antitumor activity of paclitaxel derivatives is superior to that of paclitaxel at the cellular level and in mouse models. Furthermore, some paclitaxel derivatives exhibit superior anticancer activity in paclitaxel-resistant cell models, contributing to the discovery of novel taxane drugs.

[0060] Example 1: Synthesis of Bacardine Diazo Compounds (Compound 1)

[0061]

[0062] Step 1: Add 7-O-(triethylsilane)bacasine III (3.5 mmol, 1.0 equiv) to a 250 mL round-bottom flask, dissolve it in 40 mL of anhydrous dichloromethane, and cool the reaction system to 0 °C using an ice-water bath. At this temperature, slowly add diketene (8.8 mmol, 2.5 equiv), and after the addition is complete, add DIPEA (1.0 mmol, 0.3 equiv). Stir the reaction system overnight at room temperature. After the reaction is complete as detected by TLC, concentrate the solution. Purify the crude product by column chromatography (PE:EA = 5:1) to obtain a white solid product.

[0063] Step 2: In a 250 mL round-bottom flask, add the product from Step 1 (2.5 mmol, 1.0 equiv) and p-ABSA (3.0 mmol, 1.2 equiv), dissolve them in 30 mL of acetonitrile, and cool to 0 °C in an ice-water bath. At this temperature, add DBU (3.8 mmol, 1.5 equiv) dropwise. After the addition is complete, stir overnight at room temperature. After the reaction is confirmed to be complete by TLC, quench the reaction with saturated ammonium chloride solution, extract three times with ethyl acetate, combine the organic phases, dry with Na2SO4, filter and concentrate. The crude product is purified by column chromatography (PE:EA = 10:1) to obtain a white solid product.

[0064] Step 3: Add the product from Step 2 (2.0 mmol, 1.0 equiv) to a 250 mL round-bottom flask, dissolve it in 40 mL of dichloromethane, and cool to 0 °C in an ice-water bath. At this temperature, slowly add tetrahydropyrrole (60 mmol, 30 equiv). React at room temperature for 5 hours. After the reaction is complete as detected by TLC, concentrate the product. Purify the crude product by column chromatography (PE:EA = 10:1) to obtain bacasine diazo compound 1 (white solid, 1.3 g, overall yield 49%). Spectral data of compound 1: 1 H NMR (500MHz, CDCl3) δ8.14-7.99(m,2H),7.65-7.53(m,1H),7.52-7.39(m,2H),6.44(s,1H),6.07(t,J=8.6Hz,1H ),5.65(d,J=7.0Hz,1H),4.97-4.82(m,2H),4.44(dd,J=10.5,6.7Hz,1H),4.28(d,J=8.3Hz,1H),4.14(d,J=8.4Hz ,1H),3.79(d,J=7.0Hz,1H),2.59-2.45(m,1H),2.41-2.31(m,1H),2.28(s,3H),2.22-2.13(m,4H),2.06(s,3H), 1.92-1.82(m,1H),1.76(s,1H),1.67(s,3H),1.21(s,3H),1.15(s,3H),0.91(t,J=7.9Hz,9H),0.69-0.49(m,6H).

[0065] Example 2: Synthesis of taxane derivatives (compounds 5-17)

[0066]

[0067] Add Rh2(OAc)4 (5.0 mol%) and Ni(OTf)2 (10 mol%) to a dry 5 mL test tube. Molecular sieve (100 mg) and anhydrous trifluorotoluene (1.0 mL) were sealed with a rubber stopper and stirred at room temperature under inert gas protection. Bacardi diazo compound 1 (0.1 mmol, 1.0 equiv), dimethyl tert-butylsilanol 2 (0.6 mmol, 6.0 equiv), and imine 3 (0.3 mmol, 3.0 equiv) were dissolved in 1.5 mL of trifluorotoluene, drawn into a syringe, and slowly injected using a peristaltic pump at a rate of 1.5 mL / h. After the addition was complete, the reaction was continued at room temperature for 30 minutes. After the reaction was confirmed to be complete by TLC, the insoluble matter was removed by filtration. The filtrate was concentrated and subjected to column chromatography (PE:EA = 3:1) to obtain a crude product containing multiple isomers. Further purification by reverse preparative chromatography (MeCN:H2O = 95:5) yielded protected paclitaxel derivatives. The protecting group was then removed by pyridine hydrofluoric acid to obtain compound (I'), which includes compounds 5-17.

[0068] Spectral data of compound 5: 1 H NMR (500MHz, CDCl3) δ8.20-8.07(m,2H),7.80-7.67(m,4H),7.61(t,J=7.4Hz,1H),7.56-7.45(m,3H),7.43-7.34(m,2H),7.26-7.19(m,2 H),7.06(d,J=9.0Hz,1H),6.28(s,1H),6.22(t,J=8.9Hz,1H),5.74(dd,J=9.0,2.4Hz,1H),5.67(d,J=7.0Hz,1H),4.94(dd,J=9.7,2.3Hz, 1H),4.76(s,1H),4.44-4.34(m,1H),4.30(d,J=8.5Hz,1H),4.20(d,J=8.4Hz,1H),3.92-3.82(m,1H),3.79(d,J=7.0Hz,1H),2.60-2.48( m,2H),2.38(s,3H),2.36-2.25(m,2H),2.23(s,3H),1.98(s,1H),1.90-1.85(m,1H),1.81(s,3H),1.68(s,3H),1.22(s,3H),1.14(s,3H).

[0069] Spectral data of compound 6: 1H NMR (500MHz, CDCl3) δ8.17-8.10(m,2H),7.75-7.69(m,2H),7.60(t,J=7.4Hz,1H),7.53-7.45(m,3H),7.45-7.34(m,6H),7.03(d,J=9.0Hz ,1H),6.28(s,1H),6.20(t,J=8.9Hz,1H),5.83-5.72(m,1H),5.68(d,J=7.1Hz,1H),4.94(dd,J=9.7,2.2Hz,1H),4.79(dd,J=4.9,2.4Hz,1 H),4.46-4.34(m,1H),4.30(d,J=8.5Hz,1H),4.20(d,J=8.5Hz,1H),3.80(d,J=7.0Hz,1H),3.75(d,J=5.4Hz,1H),2.60-2.49(m,2H),2.40 (s,3H),2.39-2.27(m,2H),2.22(s,3H),2.13(s,1H),1.88-1.83(m,1H),1.80(s,3H),1.68(s,3H),1.31(s,9H),1.23(s,3H),1.14(s,3H).

[0070] Spectral data of compound 7: 1 H NMR (500MHz, CDCl3) δ8.20-8.08(m,2H),7.74-7.64(m,3H),7.60(t,J=7.4Hz,1H),7.55-7.43(m,4H),7.42-7.28(m,4H),6.90(d, J=8.6Hz,1H),6.36-6.20(m,2H),6.11(dd,J=8.7,2.2Hz,1H),5.67(d,J=7.2Hz,1H),4.92(dd,J=9.6,2.3Hz,1H),4.80(dd,J=4.9 ,2.3Hz,1H),4.47-4.34(m,1H),4.28(d,J=8.4Hz,1H),4.21(d,J=8.5Hz,1H),3.92(d,J=5.9Hz,1H),3.80(d,J=7.1Hz,1H),2.60- 2.47(m,2H),2.46-2.37(m,2H),2.28(s,3H),2.23(s,3H),2.15(s,1H),1.92-1.88(m,4H),1.68(s,3H),1.25(s,3H),1.14(s,3H).

[0071] Spectral data of compound 8: 11H NMR (500 MHz, CDCl3) δ 8.23 - 8.10 (m, 2H), 7.78 - 7.68 (m, 2H), 7.62 (t, J = 7.4 Hz, 1H), 7.57 - 7.45 (m, 3H), 7.45 - 7.33 (m, 2H), 7.15 - 6.89 (m, 3H), 6.29 (s, 1H), 6.24 (t, J = 8.9 Hz, 1H), 5.96 (dd, J = 9.0, 2.2 Hz, 1H), 5.68 (d, J = 7.0 Hz, 1H), 4.95 (dd, J = 9.6, 2.2 Hz, 1H), 4.74 (dd, J = 4.8, 2.3 Hz, 1H), 4.47 - 4.36 (m, 1H), 4.31 (d, J = 8.5 Hz, 1H), 4.20 (d, J = 8.5 Hz, 1H), 3.98 (d, J = 5.0 Hz, 1H), �.81 (d, J = 7.0 Hz, 1H), 2.61 - 2.51 (m, 2H), 2.41 (s, 3H), 2.39 - 2.27 (m, 2H), 2.23 (s, 3H), 1.95 (s, 1H), 1.92 - 1.87 (m, 1H), 1.85 (s, 3H), 1.69 (s, 3H), 1..23 (s, 3H), 1.15 (s, 3H).

[0072] Spectral data of compound 9: 1 1H NMR (500 MHz, CDCl3) δ 8.18 - 8.10 (m, 2H), 7.77 - 7.70 (m, 2H), 7.65 - 7.58 (m, 1H), 7.54 - 7.46 (m, 3H), 7.43 - 7.36 (m, 2H), 7.23 - 7.13 (m, 3H), 6.96 (d, J = 8.8 Hz, 1H), 6.28 (s, 1H), 6.24 - 6.15 (m, 1H), 5.75 - 5.63 (m, 2H), 4.94 (dd, J = 9.7, 2.3 Hz, 1H), 4.75 (t, J = 3.4 Hz, 1H), 4.44 - 4.36 (m, 1H), 4.30 (d, J = 8.5 Hz, 1H), 4.19 (d, J = 8.5 Hz, 1H), 3.79 (d, J = 7.0 Hz, 1H), 3.71 (d, J = 5.4 Hz, 1H), 2.60 - 2.49 (m, 2H), 2.39 (s, 3H), 2.34 (dd, J = 15.5, 9.0 Hz, 1H), 2.28 (s, 3H), 2.27 - 2.25 (m, 1H), 2.24 - 2.20 (m, 6H), 2.00 (s, 1H), 1.85 - 1.79 (m, 4H), 1.68 (s, 3H), 1.23 (s, 3H), 1.14 (s, 3H).

[0073] Spectral data of compound 10: 1 H NMR (500MHz, CDCl3) δ8.22-8.08(m,2H),7.66(d,J=8.4Hz,2H),7.65-7.58(m,1H),7.55-7.47(m,2H),7.34(d,J=8.1Hz,4H),7.21(d,J=7.9Hz ,2H),6.98(dd,J=9.0,2.7Hz,1H),6.27(s,1H),6.20(t,J=9.0Hz,1H),5.72(dd,J=8.8,2.6Hz,1H),5.67(d,J=7.1Hz,1H),4.94(dd,J=9.5,2.3 Hz,1H),4.76(d,J=2.7Hz,1H),4.39(dd,J=10.9,6.7Hz,1H),4.30(d,J=8.5Hz,1H),4.19(d,J=8.5Hz,1H),3.79(d,J=7.0Hz,1H),3.70(s,1H), 2.61-2.47(m,2H),2.38(s,3H),2.38-2.29(m,6H),2.29-2.17(m,4H), 1.93-1.83(m,2H),1.80(s,3H),1.68(s,3H),1.23(s,3H),1.14(s,3H).

[0074] Spectral data of compound 11: 1 H NMR (500MHz, CDCl3) δ8.13(d,J=7.7Hz,2H),7.64-7.56(m,3H),7.55-7.45(m,4H),7.34(d,J=7.8Hz,2H),7.21(d,J=7.7Hz,2H),6.9 8(d,J=8.9Hz,1H),6.27(s,1H),6.20(t,J=9.0Hz,1H),5.77-5.69(m,1H),5.67(d,J=7.0Hz,1H),4.94(d,J=9.5Hz,1H),4.76(d,J=2 .6Hz,1H),4.39(dd,J=10.9,6.8Hz,1H),4.30(d,J=8.5Hz,1H),4.19(d,J=8.5Hz,1H),3.79(d,J=7.0Hz,1H),3.68(s,1H),2.62-2.4 9(m,2H),2.38(s,3H),2.36-2.29(m,4H),2.28-2.18(m,4H),1.93-1.84(m,2H),1.80(s,3H),1.68(s,3H),1.23(s,3H),1.14(s,3H).

[0075] Spectral data of compound 12: 1 H NMR (400MHz, DMSO-d6) δ7.98(d,J=7.7Hz,2H),7.68(t,J=7.4Hz,1H),7.58(t,J=7.6Hz,2H),7.30–7.19(m,3H ),6.89(d,J=8.5Hz,2H),5.94–5.71(m,2H),5.43(d,J=7.1Hz,1H),5.11(s,2H),5.01–4.63(m,3H),4.49(s,1 H),4.32(d,J=6.2Hz,1H),4.04(d,J=8.5Hz,3H),3.70(d,J=7.1Hz,1H),3.60(s,3H),2.26(s,4H),2.05–1.97 (m,1H),1.91–1.80(m,1H),1.75(s,3H),1.71–1.62(m,1H),1.53(s,3H),1.34(s,9H),1.01(d,J=9.6Hz,6H).

[0076] Spectral data of compound 13: 1 H NMR (500MHz, DMSO-d6) δ8.10(d,J=7.3Hz,2H),7.58(t,J=7.4Hz,1H),7.47(t,J=7.7Hz,2H),6.87–6.78(m,3H),6.20(t,J=9.0Hz ,1H),5.67(d,J=7.1Hz,1H),5.45(d,J=9.5Hz,1H),5.15(s,2H),4.93(d,J=9.3Hz,1H),4.52(s,1H),4.38–4.33(m,1H),4.30(d,J =8.5Hz,1H),4.17(d,J=8.5Hz,1H),3.89(s,4H),3.86(s,3H),2.51(s,4H),2.39–2.32(m,1H),2.21–2.11(m,1H),1.86(s,4H),1 .71(s,1H),1.67(s,3H),1.34(s,9H),1.20(d,J=3.9Hz,6H),0.81(t,J=8.0Hz,9H),0.49–0.36(m,6H),0.17(s,9H),0.11(s,9H).

[0077] Spectral data of compound 14: 1HNMR(500MHz,DMSO-d6)δ8.01–7.98(m,2H),7.58–7.51(m,1H),7.48–7.40(m,2H),7.40–7.33(m,2H),7.35–7.29(m,1H),7.2 9–7.24(m,1H),6.84(d,J=8.1Hz,1H),5.44(d,J=7.2Hz,1H),5.43(d,J=7.2Hz,1H),5.13(d,J=2.6Hz,1H),5.10(d,J=7.2Hz, 1H),4.95(d,J=2.5Hz,1H),4.90–4.85(m,2H),4.50(s,1H),4.34(m,1H),4.09–3.98(m,3H),3.67(d,J=7.2Hz,1H),2.32–2.2 3(m,1H),2.21(s,3H),1.95–1.85(m,1H),1.80–1.62(m,5H),1.53(s,3H),1.37(s,8H),1.25(s,1H),1.01(d,J=10.6Hz,6H).

[0078] Spectral data of compound 15: 1 HNMR(500MHz,DMSO-d6)δ8.01–7.94(m,2H),7.56–7.50(m,1H),7.48–7.41(m,2H),7.45–7.36(m,1H),7.36–7.25(m,3H ),6.58–6.48(m,2H),5.46(d,J=7.2Hz,1H),5.40(d,J=7.2Hz,1H),5.12(d,J=2.6Hz,1H),5.08(d,J=7.2Hz,1H),4.95( d,J=2.5Hz,1H),4.90–4.80(m,2H),4.50(s,1H),4.35(m,1H),4.10–3.99(m,3H),3.68(d,J=7.2Hz,1H),2.32–2.21(m, 1H),2.21(s,3H),1.97–1.86(m,1H),1.78–1.63(m,5H),1.53(s,3H),1.37(s,8H),1.25(s,1H),1.00(d,J=9.6Hz,6H).

[0079] Spectral data of compound 16: 1HNMR(500MHz,DMSO-d6)δ8.02–7.96(m,2H),7.57–7.52(m,1H),7.49–7.42(m,4H),7.33–7.27(m,2H),6.81(d,J=8 .1Hz,1H),5.48(d,J=7.2Hz,1H),5.42(d,J=7.2Hz,1H),5.13(d,J=2.6Hz,1H),5.10(d,J=7.2Hz,1H),4.93(d,J=2. 5Hz,1H),4.92–4.83(m,2H),4.50(s,1H),4.35(m,1H),4.10–3.98(m,3H),3.68(d,J=7.2Hz,1H),2.32–2.21(m,1H) ,2.21(s,3H),1.97–1.86(m,1H),1.79–1.63(m,5H),1.54(s,3H),1.36(s,8H),1.26(s,1H),1.01(d,J=9.6Hz,6H).

[0080] Spectral data of compound 17: 1 H NMR (500MHz, DMSO-d6) δ8.00(d,J=7.7Hz,2H),7.67(t,J=7.4Hz,1H),7.57(t,J=7.6Hz,2H),7.14(d,J=9.6Hz,1H),6.12(d,J =3.1Hz,1H),6.00(d,J=3.0Hz,1H),5.96–5.87(m,2H),5.46(d,J=7.1Hz,1H),5.13(d,J=2.6Hz,1H),5.07(d,J=7.2Hz,1H),5 .00–4.89(m,3H),4.65(s,1H),4.45–4.38(m,1H),4.09–4.02(m,3H),3.74(d,J=7.0Hz,1H),2.31(s,4H),2.22(s,3H),2.18– 2.13(m,1H),2.09(t,J=7.8Hz,1H),1.79–1.74(m,3H),1.70–1.63(m,1H),1.54(s,3H),1.36(s,9H),1.02(d,J=11.8Hz,6H).

[0081] The structure of compound 5-17 is as follows:

[0082]

[0083] Example 3: Determination of the inhibitory effect of paclitaxel derivatives on tumor cell growth in an in vitro model.

[0084] (1) CCK-8 assay for cell viability

[0085] 1) Cell culture:

[0086] In Example 3 of this invention, the MDA-MB-231, MCF-7, SKOVA3, and HepG2 cells (purchased from ATCC) were cultured in DMEM medium containing 10% fetal bovine serum and 1% penicillin and streptomycin; the KYSE-520 and 4T1 cells (purchased from ATCC) were cultured in RPIM-1640 medium containing 10% fetal bovine serum and 1% penicillin and streptomycin; and the A549 cells (purchased from ATCC) were cultured in Ham's medium containing 10% fetal bovine serum and 1% penicillin and streptomycin. Paclitaxel-resistant human colon cancer HCT15 cells were cultured and preserved in our laboratory in RPIM-1640 medium containing 10% fetal bovine serum and 1% penicillin and streptomycin (paclitaxel concentration 10 mg / ml). Paclitaxel-resistant human non-small cell lung cancer A549 cells were also cultured and preserved in our laboratory in Ham's F12K medium containing 10% fetal bovine serum and 1% penicillin and streptomycin (paclitaxel concentration 10 mg / ml). All cells were cultured at 37°C in a 5% cell culture incubator.

[0087] 2) Compound activity testing:

[0088] ① Seed 200 μL of cell suspension (5000 cells / well) into a 96-well plate and incubate the plate at 37°C for 24 h in a 5% cell culture incubator. The compound prepared in Example 2 of this invention was dissolved in DMSO, and different concentrations of the compound prepared in Example 2 of this invention were added to 1 μL of the culture plate, and incubated for another 48 h.

[0089] ② Discard the culture medium and add fresh culture medium containing 10% CCK-8. After incubating at 37°C and 5% CO2 for 2 hours, measure the absorbance at 450 nm.

[0090] 3) Experimental results:

[0091] Table 1. Determination of the inhibitory effects of paclitaxel derivatives on the growth of different tumor cells.

[0092]

[0093] Table 2. Determination of the inhibitory effects of docetaxel derivatives on the growth of different tumor cells.

[0094]

[0095]

[0096] As shown in Tables 1 and 2, both paclitaxel and docetaxel derivatives have inhibitory effects on a variety of tumor cells.

[0097] Example 4: Study on the inhibitory effect of paclitaxel derivatives (compound 8) on tumor growth in an in vivo model.

[0098] (1) Effects on mouse subcutaneous breast cancer 4T1 cell xenograft model

[0099] 1) Construction of tumor mouse models and drug administration regimens

[0100] ① Take mouse breast cancer cells (4T1 breast cancer cells) in the logarithmic growth phase, digest and count them, mix pre-cooled PBS and Matrigel at a 1:1 ratio, resuspend the cells to obtain a concentration of 5 x 10⁻⁶ cells / mL. 6 A cell suspension of 1 cell / ml was placed on ice.

[0101] ② 100 μL of cell suspension was injected subcutaneously into the abdominal and dorsal sides of 4-5 week old Balb / C mice. When the subcutaneous tumor size reached 50 mm, the mice were randomly divided into four groups. The group classification and drug administration settings are as follows.

[0102] Blank control group;

[0103] Dosage group: The compound prepared in Example 2 of this invention at a concentration of 85 mg / kg;

[0104] The compound prepared in Example 2 of this invention has a concentration of 810 mg / kg.

[0105] The compound prepared in Example 2 of this invention has a concentration of 820 mg / kg.

[0106] Positive control group: Paclitaxel 20mg / Kg group.

[0107] Administration: The control group received 100 μL of placebo (i.e., the solvent used to dissolve the drug: 15% castor oil + 85% sterile PBS) intraperitoneally every two days; the other treatment groups received 100 μL of different doses of drug solution intraperitoneally every two days.

[0108] ③ During 21 days of continuous administration, the body weight and tumor size of mice were measured every two days, and a mouse body weight growth curve was plotted;

[0109] ④ After administration of the drug for 21 hours, the mice were euthanized, dissected, and the subcutaneous tumors were removed and weighed.

[0110] 2) Experimental Results

[0111] The experimental results are shown in Table 3. Table 3 shows the tumor inhibition rate after administration in a mouse subcutaneous breast cancer cell xenograft model. All three concentrations of compound 8 significantly inhibited the growth of tumors in mice, with the best effect observed at a concentration of 20 mg / kg.

[0112] Table 3. Tumor inhibition rate after drug administration in mouse subcutaneous breast cancer cell xenograft model

[0113]

[0114]

[0115] like Figure 2 As shown, experiments using a Balb / c mouse subcutaneous cell transplantation tumor model revealed that small molecule compounds derived from taxanes can effectively inhibit the growth of breast cancer cells.

[0116] Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used in this specification is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.

[0117] As used in this invention, the terms "comprising" and "including" are open-ended expressions, meaning they include the contents specified in this invention but do not exclude other aspects.

[0118] As used in this invention, the term "and / or" includes any one or more of the related listed items and all combinations thereof.

[0119] The embodiments of the present invention have been described in detail above, but the present invention is not limited to the described embodiments. For those skilled in the art, various changes, modifications, substitutions, and variations can be made to these embodiments without departing from the principles and spirit of the present invention, and these variations still fall within the protection scope of the present invention.

Claims

1. A paclitaxel derivative having the formula (I), characterized in that, Its structure is as follows: In the formula, R1 group is hydrogen, substituted silicon group, C1-C4 alkyl, -CH2-C3-C5 cycloalkyl, or C3-C8 cycloalkyl; The R2 group can be hydrogen, C1-C4 alkyl, C3-C8 cycloalkyl, C1-C4 acyl, or C3-C8 cyclocarboxyl. The R3 group can be hydrogen, substituted silicon, C1-C4 alkyl, C3-C8 cycloalkyl, C1-C4 acyl, or C3-C8 cyclocarboxyl. The R4 group is phenyl, substituted phenyl, pyridine, pyrimidine, pyrrole, pyrazole, pyridazine, furan, substituted furan, thiophene, substituted thiophene, C6-C12 fused ring, C1-C4 alkoxy, wherein the substituent is independently selected from -F, -Cl, and -Br; Ar1 group is phenyl, substituted phenyl, pyridine, pyrimidine, pyrrole, pyrazole, pyridazine, furan, substituted furan, thiophene, substituted thiophene, substituted indole, C6-C12 fused ring, wherein the substituent is independently selected from -F, -Cl, -Br, -CH3, -OCH3.

2. The paclitaxel derivative having formula (I) according to claim 1, characterized in that: The R1 group is hydrogen, methyl, ethyl, propyl, isopropyl, cyclopropyl, -CH2-cyclobutyl, tert-butyl, trimethylsilyl, triethylsilyl, tert-butyldimethylsilyl, or triphenylsilyl. The R2 group can be hydrogen, methyl, ethyl, isopropyl, cyclopropyl, formyl, acetyl, cyclopropylformyl, cyclobutylformyl, or cyclopentanoyl. The R3 group can be hydrogen, triethylsilyl, methyl, ethyl, isopropyl, cyclopropyl, formyl, acetyl, cyclopropylformyl, cyclobutylformyl, or cyclopentanoyl. The R4 group is a substituted phenyl, 2-pyridine, 3-pyridine, 4-pyridine, pyrimidine, pyrrole, pyrazole, pyridazine, 2-furan, 2-substituted furan, 3-furan, 3-substituted furan, 2-thiophene, 2-substituted thiophene, 3-thiophene, 3-substituted thiophene, C6-C12 fused ring, or tert-butoxy, wherein the substituent is independently selected from -F, -Cl, or -Br; Ar1 group is phenyl, substituted phenyl, 2-pyridine, 3-pyridine, 4-pyridine, pyrimidine, pyrrole, pyrazole, pyridazine, 2-furan, 2-substituted furan, 3-furan, 3-substituted furan, 2-thiophene, 2-substituted thiophene, 3-thiophene, 3-substituted thiophene, substituted indole, C6-C12 fused ring, wherein the substituent is independently selected from -F, -Cl, -Br, -CH3, -OCH3.

3. The paclitaxel derivative having formula (I) according to claim 1 or 2, characterized in that, The taxol derivatives are selected from compounds with the following structures:

4. A method for preparing a paclitaxel derivative having formula (I) as described in any one of claims 1-3, characterized in that, The method includes the following steps: preparing a mixed solution a by dissolving the compounds shown in Formulas 1, 2, and 3 in an organic solvent according to the following reaction formulas; and adding a metal catalyst, additives, and... Molecular sieves are dispersed in the organic solvent to obtain mixture b; then, mixed solution a is added to mixture b to obtain a paclitaxel derivative with a protecting group, and finally, under the action of pyridine hydrogen fluoride, a paclitaxel derivative of formula (I) is obtained. The reaction route is shown below: Among them, R1 group is hydrogen, substituted silicon group, C1-C4 alkyl, -CH2-C3-C5 cycloalkyl, C3-C8 cycloalkyl; The R2 group can be hydrogen, C1-C4 alkyl, C3-C8 cycloalkyl, C1-C4 acyl, or C3-C8 cyclocarboxyl. The R3 group can be hydrogen, substituted silicon, C1-C4 alkyl, C3-C8 cycloalkyl, C1-C4 acyl, or C3-C8 cyclocarboxyl. The R4 group is phenyl, substituted phenyl, pyridine, pyrimidine, pyrrole, pyrazole, pyridazine, furan, substituted furan, thiophene, substituted thiophene, C6-C12 fused ring, C1-C4 alkoxy, wherein the substituent is independently selected from -F, -Cl, and -Br; Ar1 group is phenyl, substituted phenyl, pyridine, pyrimidine, pyrrole, pyrazole, pyridazine, furan, substituted furan, thiophene, substituted thiophene, substituted indole, C6-C12 fused ring, wherein the substituent is independently selected from -F, -Cl, -Br, -CH3, -OCH3.

5. The preparation method according to claim 4, characterized in that, The reaction molar ratio of the compound shown in Formula 1, the compound shown in Formula 2, the compound shown in Formula 3, the metal catalyst, and the additive is (0.5-2):(1-6):(1-4):(0.05-0.3):(0.1-0.5).

6. The preparation method according to claim 4 or 5, characterized in that, The metal catalyst comprises one or more of Rh2(OAc)4, Rh2(oct)4, Rh2(esp)2, Rh2(Piv)4, Cu(OTf)2, Pd(ally)2Cl2, and Cu(MeCN)4BF4; and / or the additive comprises AgPF6, AgBF4, AgOTf, AgTFA, AgClO4, AgSbF6, AgOTf, Cu(OTf)2, Mg(OTf)2, Sc(OTf)3, Fe(OTf)2, and Yb(OTf).

2. One or more of La(OTf)2, Bi(OTf)3, In(OTf)2, LiOTf, Zn(OTf)2, Hg(OTf)2, Ni(OTf)2, NaOTf, and Ca(OTf)2; and / or, the organic solvent includes one or more of dichloromethane, chloroform, ethyl acetate, tetrahydrofuran, toluene, xylene, chlorobenzene, fluorobenzene, trifluorotoluene, and 1,4-dioxane; and / or, the reaction temperature is -20℃ to 40℃; and / or, the reaction time is 0.5-4 hours.

7. A pharmaceutical composition, characterized in that, Includes a paclitaxel derivative of formula (I) as described in any one of claims 1-3 and a pharmaceutically acceptable carrier.

8. The use of any one of the paclitaxel derivatives of formula (I) according to any one of claims 1-3, or the pharmaceutical composition according to claim 7, in the preparation of an antitumor drug.

9. The use of any one of the paclitaxel derivatives of formula (I) according to any one of claims 1-3 or the pharmaceutical composition according to claim 7 in the preparation of a medicament for inhibiting tumor cell proliferation.

10. The application according to claim 8 or 9, characterized in that, The tumor cells include triple-negative breast cancer, ovarian cancer, lung cancer, colorectal cancer, melanoma, head and neck cancer, lymphoma, and brain tumors.

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