PPD ring-opening epoxy substituted derivative as well as preparation method and application thereof

Abstract

The invention discloses a PPD ring-opening epoxy substituted derivative, the structural formula of which is as shown in a formula 8 or a formula 9, or pharmaceutically acceptable salts of compounds as shown in the formula 8 and the formula 9. The invention discloses a PPD ring-opening epoxy substituted derivative, and discloses a preparation method of the PPD ring-opening epoxy substituted derivative, and application of the PPD ring-opening epoxy substituted derivative and pharmaceutically acceptable salts thereof in preparation of a tumor drug resistance reversal agent / tumor drug sensitizer, or preparation of a tumor drug resistance reversal agent / tumor drug sensitizer containing a pharmaceutically acceptable carrier. The PPD ring-opening epoxy substituted derivative disclosed by the invention is synthesized for the first time, the anti-tumor MDR activity of the derivative is found for the first time, the cytotoxicity is low, the PPD ring-opening epoxy substituted derivative 8a-8b and 9a-9h has better tumor drug resistance reversal activity due to covalent crosslinking groups, and the effect is obviously superior to that of a positive control drug. # imgabs0 #

Description

Technical Field

[0001] The invention relates to the fields of organic synthesis and medicinal chemistry, and in particular to a novel PPD ring-opening epoxy substituted derivative, a preparation method thereof and an application thereof in the field of medicine. Background Art

[0002] Multidrug resistance (MDR) is an important cause of failure of tumor chemotherapy. The generation of tumor MDR makes it impossible for anti-tumor drugs to stay in tumor cells, making them lose their therapeutic effects, which poses a severe challenge to current tumor treatment and drug development. However, existing multidrug resistance reversal agents have problems such as low efficacy and high toxicity. Therefore, the development of multidrug resistance reversal agents with new structures, low toxicity and high efficiency is a hot topic in tumor treatment and drug research.

[0003] Ginseng is known as the "king of herbs" and is a widely used Chinese herbal medicine. Ginsenosides, as the effective active ingredients in ginseng, have multiple pharmacological activities such as antioxidant, anti-inflammatory, antibacterial, anti-cancer and anti-myocardial ischemia. Among them, 20S-protopanaxadiol (PPD), as the main sapogenin, is now commonly used as an auxiliary drug during the treatment of cancer patients. 20S-PPD is easily metabolized into a structure with a furan ring as the side chain in the body, and its 3-hydroxyl group is also very susceptible to liver metabolism. Therefore, if the PPD side chain is selected to form a furan ring, and a series of derivatives that are not easily metabolized are constructed by opening the A ring, it is possible to obtain a new P-gp inhibitor with strong activity and low toxicity. Summary of the invention

[0004] In order to design a novel PPD derivative with novel structure and the ability to reverse the tumor MDR activity mediated by P-glycoprotein, and further provide a drug for treating tumors and its composition. The present invention provides a series of derivatives with structural features shown in formula A, B, and pharmaceutically acceptable salts thereof. The anti-tumor MDR activity of these compounds is discovered for the first time, and the compounds are synthesized for the first time.

[0005] The technical solution adopted by the present invention is:

[0006] A PPD ring-opening epoxy substituted derivative, whose structural formula is shown in Formula 8 or Formula 9, or a pharmaceutically acceptable salt of the compound shown in Formula 8 or Formula 9,

[0007]

[0008] Wherein, R1 and R2 are each independently R x -KY-, where Y represents NH or O, K represents methylene or is absent, R xis a C3-C10 heterocyclic or heteroaromatic group which is unsubstituted or substituted with one or more substituents; the substituent is a C1-C10 alkyl group or tert-butyloxycarbonyl group. The heterocyclic or heteroaromatic group of the present invention refers to a heterocyclic or heteroaromatic group containing one or both of N and O atoms in the ring.

[0009] Preferred R x It is one of pyrazine, pyridine, azetidine, thiophene, piperidine, furan, tetrahydrofuran, and thiazole.

[0010] Furthermore, preferably, R1 represents 1-methyl-(2-thiophene)-1-yl or 3-N-Boc-azetidine-1-yl; R2 represents 4-methylpiperidin-1-yl, 1-methyl-(2-furan)-1-yl, 1-methyl-(2-thiazole)-1-yl, 1-methyl-(2-thiophene)-1-yl, 3-N-Boc-azetidine-1-yl, 4-N-Boc-piperidin-1-yl, 3-oxetan-1-yl, or 1-methyl-(2-pyridine)-1-yl.

[0011] More preferably, the compound represented by Formula 8 or Formula 9 is one of the following

[0012]

[0013] The names of the compounds are as follows:

[0014] (20S,24R)-epoxy-3,4-opened-3-(2-thienylmethylamine-1-yl)carbonyl-4(28)-oxirane-dammarane-12β,25-diol (Compound 8a);

[0015] (20S,24R)-epoxy-3,4-opened-3-(3-N-Boc-azetidinamine-1-yl)carbonyl-4(28)-oxirane-dammarane-12β,25-diol (Compound 8b);

[0016] (20S,24R)-epoxy-3,4-opened-3-(4-methylpiperidinol-1-yl)carbonyl-4(28)-oxirane-dammarane-12β,25-diol (Compound 9a);

[0017] (20S,24R)-epoxy-3,4-opened-3-(furfuryl-1-yl)carbonyl-4(28)-oxirane-dammarane-12β,25-diol (Compound 9b);

[0018] (20S,24R)-epoxy-3,4-opened-3-(2-thiazolylmethanol-1-yl)carbonyl-4(28)-oxirane-dammarane-12β,25-diol (Compound 9c);

[0019] (20S,24R)-epoxy-3,4-opened-3-(2-thiophenecarbinol-1-yl)carbonyl-4(28)-oxirane-dammarane-12β,25-diol (Compound 9d);

[0020] (20S,24R)-epoxy-3,4-opened-3-(3-N-Boc-azetidinol-1-yl)carbonyl-4(28)-oxirane-dammarane-12β,25-diol (Compound 9e);

[0021] (20S,24R)-epoxy-3,4-opened-3-(4-N-Boc-piperidinol-1-yl)carbonyl-4(28)-oxirane-dammarane-12β,25-diol (Compound 9f);

[0022] (20S,24R)-epoxy-3,4-opened-3-(3-oxetanol-1-yl)carbonyl-4(28)-oxirane-dammarane-12β,25-diol (Compound 9g);

[0023] (20S,24R)-Epoxy-3,4-seco-3-(2-pyridiniummethanol-1-yl)carbonyl-4(28)-oxirane-dammarane-12β,25-diol (Compound 9h).

[0024] More preferably, the PPD ring-opening epoxy substituted derivative is compound 8a, 8b, 9c, 9d, 9e, 9f.

[0025] The present invention also provides a PPD ring-opened epoxy substituted derivative shown in Formula 8 or Formula 9 and a pharmaceutically acceptable salt thereof, and use thereof in the preparation of a tumor resistance reversal agent / tumor drug sensitizer, or the preparation of a tumor resistance reversal agent / tumor drug sensitizer containing a pharmaceutically acceptable carrier; further, the PPD ring-opened epoxy substituted derivative shown in Formula 8 or Formula 9 and a pharmaceutically acceptable salt thereof can be used to prepare a tumor resistance reversal agent / tumor drug sensitizer that overcomes P-glycoprotein-mediated tumor resistance, or to prepare a tumor resistance reversal agent / tumor drug sensitizer containing a pharmaceutically acceptable carrier.

[0026] Furthermore, in the application, the PPD ring-opening epoxy substituted derivative is compound 8a, 8b, 9c, 9d, 9e, 9f.

[0027] The present invention also provides the use of the PPD ring-opening epoxy substituted derivatives shown in Formula 8 or Formula 9 and pharmaceutically acceptable salts thereof in the preparation of anti-tumor drugs. In the method of application, the anti-tumor drugs include clinical anti-tumor drugs and tumor resistance reversal agents / tumor drug sensitizers, and the tumor resistance reversal agents / tumor drug sensitizers are PPD ring-opening epoxy substituted derivatives shown in Formula 8 or Formula 9 and pharmaceutically acceptable salts thereof.

[0028] Furthermore, the tumor is breast cancer, colon cancer, cervical cancer, liver cancer, gastric cancer or lung cancer.

[0029] The clinical anti-tumor drug is paclitaxel.

[0030] Furthermore, in the application, the PPD ring-opening epoxy substituted derivative is preferably compound 8a, 8b, 9c, 9d, 9e, 9f.

[0031] The present invention also provides the use of the PPD ring-opening epoxy substituted derivatives shown in Formula 8 or Formula 9 and pharmaceutically acceptable salts thereof in the preparation of auxiliary drugs or health products for tumor patients during chemotherapy, and the application is not limited to the treatment and prevention of tumor multidrug resistant diseases.

[0032] The pharmaceutically acceptable salts described in the present invention refer to conventional acid addition salts, which have the same pharmaceutical efficacy as the compounds and are salts formed with suitable non-toxic organic or inorganic acids.

[0033] The present invention also provides a pharmaceutical composition for resisting multidrug resistance, which comprises a PPD open-ring epoxy-substituted derivative shown in Formula 8 or Formula 9 and a pharmaceutically acceptable salt thereof, and can also be added with a pharmaceutically acceptable carrier to prepare a pharmaceutical preparation such as tablets, capsules, powders, syrups, suspensions, injections, etc.; and can also be added with commonly used pharmaceutical excipients such as sweeteners, diluents, and fillers.

[0034] Furthermore, in the pharmaceutical composition, the PPD ring-opening epoxy substituted derivative is preferably compound 8a, 8b, 9c, 9d, 9e, 9f.

[0035] The pharmaceutical composition provided by the present invention can be administered clinically by oral administration, injection or the like.

[0036] The clinical dosage of the pharmaceutical composition of the present invention is 0.01 mg to 1000 mg / day of the active component, and may deviate from this range depending on the severity of the disease or the dosage form.

[0037] Furthermore, the present invention also provides a method for preparing a ring-opening epoxy-substituted derivative of PPD shown in Formula 8 or Formula 9, and the method is shown in the following reaction formula:

[0038]

[0039] The method comprises the following steps:

[0040] (1) 24R-Pyxinol shown in Formula 1 and pyridinium chlorochromate (PCC) are added in a molar ratio of 1:1 to 8 to carry out an oxidation reaction to obtain compound 3;

[0041] The reaction solvent of step (1) is an aprotic solvent, preferably dichloromethane (DCM) or tetrahydrofuran;

[0042] The reaction temperature of the oxidation reaction is -20°C to 60°C; preferably, the reaction temperature is 0°C to 30°C, and more preferably, room temperature.

[0043] The reaction time of the oxidation reaction is 3 to 24 hours, preferably 3 to 12 hours. TLC is usually used to monitor the reaction until the reaction is complete.

[0044] After the reaction is completed, the reaction solution is post-treated to obtain compound 3. The post-treatment method of the reaction solution is: filtering the reaction solution, vacuum concentrating, and then purifying by silica gel column chromatography (SGCC) to obtain compound 3.

[0045] (2) Compound 3 is subjected to Baeyer-Villiger reaction with m-chloroperbenzoic acid (m-CPBA) to obtain compound 4;

[0046] The molar ratio of compound 3 to m-chloroperbenzoic acid (m-CPBA) is 1:3-15.

[0047] Further, the reaction solvent of step (2) is an aprotic solvent, preferably dichloromethane (DCM) or tetrahydrofuran;

[0048] The reaction temperature of the Baeyer-Villiger reaction is -10°C to 40°C, preferably 5°C to 30°C, more preferably room temperature; the reaction time is 5 to 12 hours.

[0049] After the reaction is completed, the obtained reaction solution is post-treated to obtain compound 4. The post-treatment method of the reaction solution is: the reaction solution is stirred, filtered, concentrated in vacuo, and then purified by silica gel column chromatography (SGCC) to obtain compound 4.

[0050] (3) Compound 4 is subjected to an acidic ring-opening reaction with p-toluenesulfonamide (P-TSA) to obtain compound 5;

[0051] The molar ratio of the compound 4 to p-toluenesulfonamide (P-TSA) is 1:3-15.

[0052] Further, the reaction solvent of step (3) is an aprotic solvent, preferably dichloromethane (DCM) or tetrahydrofuran;

[0053] The reaction temperature of the acidic ring-opening reaction is -10°C to 60°C, preferably 5°C to 30°C, more preferably room temperature; the reaction time is 5 to 18 hours.

[0054] After the reaction is completed, the obtained reaction solution is post-treated to obtain compound 5. The post-treatment method of the reaction solution is: the reaction solution is stirred, filtered, concentrated in vacuo, and then purified by silica gel column chromatography (SGCC) to obtain compound 5.

[0055] (4) subjecting compound 5 to an amide reaction or an esterification reaction with an amino-containing heterocyclic compound R1-NH2 or a hydroxyl-containing heterocyclic compound R2-OH to obtain compound 6 or compound 7, respectively;

[0056] The molar ratio of the compound 5 to the amino-containing heterocyclic compound R1-NH2 or the hydroxyl-containing heterocyclic compound R2-OH is 1:1.2-16.

[0057] Furthermore, the catalyst for the amide reaction is HBTU (benzotriazole-N,N,N,N-tetramethyluronium hexafluorophosphate) and triethylamine, and the molar ratio of compound 5 to HBTU and triethylamine is 1:1.3-5:2-10;

[0058] Furthermore, the catalysts for the esterification reaction are EDCI and DMAP, and the molar ratio of compound 5 to EDCI and DMAP is 1:2-5:0.1-1.

[0059] Further, the reaction solvent of step (4) is an aprotic solvent, preferably N,N-dimethylformamide or dichloromethane (DCM);

[0060] The reaction temperature of the amide reaction or esterification reaction is -10°C to 40°C, preferably 0 to 30°C, more preferably room temperature; the reaction time is 0.5 to 16 hours, preferably 0.5 to 10 hours.

[0061] After the reaction is completed, the reaction solution is post-treated to obtain compound 6 or compound 7, respectively. The post-treatment method of the reaction solution is: adding water to the reaction solution to quench the reaction, extracting with ethyl acetate, washing and drying the organic phase, filtering and concentrating it, and purifying the concentrated sample by silica gel column chromatography to obtain compound 6 or compound 7, respectively.

[0062] (5) Compound 6 or Compound 7 and m-chloroperbenzoic acid (m-CPBA) are added in a molar ratio of 1:1.2 to 12 to carry out a cyclization reaction to obtain a PPD ring-opening epoxy substituted derivative represented by Formula 8 or Formula 9, respectively;

[0063] In the cyclization reaction of step (5), sodium bicarbonate is also added, and the molar ratio of compound 6 or compound 7 to sodium bicarbonate is preferably 1:5-15.

[0064] The reaction solvent of step (5) is an aprotic solvent, preferably dichloromethane (DCM) or tetrahydrofuran;

[0065] The reaction temperature of the cyclization reaction is -10°C to 60°C; preferably, the reaction temperature is 0°C to 30°C.

[0066] The reaction time of the cyclization reaction is 3 to 24 hours; preferably, the reaction time is 4 to 12 hours.

[0067] After the reaction is completed, the reaction solution is post-treated to obtain the PPD ring-opening epoxy substituted derivatives shown in Formula 8 or Formula 9, respectively. The post-treatment method of the reaction solution is: adding water to the reaction solution to quench the reaction, extracting with ethyl acetate, filtering and concentrating the organic phase after washing and drying, and purifying the concentrated sample by silica gel column chromatography to obtain the PPD ring-opening epoxy substituted derivatives shown in Formula 8 or Formula 9, respectively.

[0068] Compared with the prior art, the present invention has the following advantages:

[0069] 1. The ring-opening epoxy-substituted derivatives of PPD and their medically acceptable salts shown in formulas 8 and 9 provided by the present invention are synthesized for the first time, and their anti-tumor MDR activity is discovered for the first time.

[0070] 2. The ring-opening epoxy-substituted derivatives of PPD represented by formula (8a-8b) and (9a-9h) of the present invention have lower cytotoxicity than the relatively low-toxic classic P-glycoprotein inhibitor verapamil (Vrp). For example, when the drug concentration is 30 μM, the survival rate of the tumor resistant cells KBV treated with Vrp is about 40%, while compounds 8a-8b and 9a-9h do not show cytotoxicity, and the survival rate is above 80%.

[0071] 3. The compounds of the present invention have better tumor resistance reversal effect. Compared with sapogenin PPD and parent core compounds 6 and 7, the PPD ring-opening epoxy substituted derivatives of the present invention have better tumor resistance reversal ability and activity in inhibiting P-glycoprotein function. For example, the commonly used clinical drug paclitaxel has no cytotoxicity to the tumor-resistant cell line KBV under the action of 100nM. When the PPD ring-opening epoxy substituted derivatives at a concentration of 10μM are added together, the survival rate of KBV cells is significantly reduced, indicating that their sensitivity to paclitaxel-resistant tumor-resistant cells KBV to paclitaxel is significantly improved, that is, it shows a tumor resistance reversal effect. And compared with PPD derivatives 6a-6b and 7a-7h without covalent cross-linking groups, PPD ring-opening epoxy substituted derivatives 8a-8b and 9a-9h that form covalent cross-linking groups through epoxy ring formation have better tumor resistance reversal activity. Among them, the tumor resistance reversal activity of preferred compounds 8a, 8b, 9c, 9d, 9e, and 9f is significantly better than that of the positive drug verapamil (Vrp). The PPD ring-opening epoxy substituted derivative of the invention has great development and application value. DETAILED DESCRIPTION

[0072] The present invention is further described below in conjunction with specific embodiments, but the protection scope of the present invention is not limited to these embodiments.

[0073] Example 1: (20S, 24R)-epoxy-3,4-opened-3-(2-thiophenemethylamine-1-yl)carbonyl-4(28)-oxirane-dammarane-12β,25-diol (Compound 8a);

[0074] Protopanaxadiol (20S-PPD) (200 mg, 0.534 mmol) was used as the raw material, dissolved in anhydrous dichloromethane (17 mL), and oxidative cyclization reaction was carried out using m-CPBA (109.866 mg, 0.736 mmol). After stirring at room temperature for 12 h, the mixture was filtered, concentrated in vacuo, and then chromatographed on a silica gel column (SGCC) to obtain intermediates 1 and 2. Intermediate 1 (82.0 mg, 0.172 mmol) was dissolved in anhydrous dichloromethane (2 mL), pyridinium chlorochromate (39.0 mg, 0.180 mmol) was added, stirred at room temperature for 3 h, filtered, concentrated in vacuo, and chromatographed on a silica gel column (SGCC) to obtain intermediate 3. The intermediate product 3 (130 mg, 0.27 mmol) was dissolved in 6 mL of dichloromethane, and m-CPBA (200 mg, 0.976 mmol) was added. After stirring at room temperature overnight, the mixture was filtered, concentrated in vacuo, and then subjected to silica gel column chromatography (SGCC) to obtain the intermediate product 4. The intermediate product 4 (220 mg, 0.45 mmol) was dissolved in anhydrous dichloromethane (9 mL), and P-TSA (341.0 mg, 1.795 mmol) was added. After stirring at room temperature for 12 h, the mixture was filtered, concentrated in vacuo, and then subjected to silica gel column chromatography (SGCC) to obtain the intermediate product 5. The intermediate product 5 (25 mg, 0.051 mmol) and 2-thiophenemethylamine (0.061 mmol) were dissolved in anhydrous DCM (1 mL), HBTU (24 mg, 0.076 mmol) and 1 drop of Et3N were added, and the mixture was stirred at room temperature for 10 min, and then an appropriate amount of deionized water was added to quench the mixture, and the organic layer was extracted with EA, and the organic layer was washed with saturated NaHCO3 solution and saturated NaCl aqueous solution in turn, and then dried over Na2SO4, concentrated, and column chromatography was performed to obtain the intermediate product 6a. The intermediate product 6a (30.0 mg) was dissolved in anhydrous DCM (4 mL), and NaHCO3 (33.0 mg, 0.409 mmol) was added, and then m-CPBA (11.4 mg, 0.066 mmol) was slowly added thereto, and the mixture was stirred at room temperature for 2 h. Subsequently, an appropriate amount of deionized water was added for quenching, the organic layer was extracted with EA, and the organic layer was washed with saturated NaHCO3 solution and saturated NaCl aqueous solution in turn, and then dried over NaS04 and concentrated. The target compound 8a [(20S, 24R)-epoxy-3,4-opened-3-(2-thienylmethylamine-1-yl)carbonyl-4(28)-oxirane-dammarane-12β,25-diol] was obtained by column chromatography.

[0075] 1H NMR (400MHz, CDCl3) δ7.22 (dd, J=4.9, 1.5Hz, 1H), 6.96 (tt, J=5.0, 2.4Hz, 2H) ,4.59(qt,J=15.2,7.8Hz,2H),3.85(dd,J=8.7,6.7Hz,1H),3.49(td,J=10.5, 4.4Hz,1H),2.96(d,J=4.1Hz,1H),2.75(d,J=4.0Hz,1H),2.22-1.31(m,22H), 1.27(s,6H),1.26(s,3H),1.10(s,3H),1.01(s,3H),0.96(s,3H),0.84(s,3H).

[0076] Example 2: (20S, 24R)-epoxy-3,4-opened-3-(3-N-Boc-azetidinamine-1-yl)carbonyl-4(28)-oxirane-dammarane-12β,25-diol (Compound 8b);

[0077] Under argon protection, the intermediate product 5 (250 mg, 0.510 mmol) in Example 1 and 3-N-Boc-azetidinamine (0.612 mmol) were dissolved in anhydrous DCM (10 mL), HBTU (240 mg, 0.76 mmol) and Et3N (150 μL) were added, and the mixture was stirred at room temperature for 10 min, and then an appropriate amount of deionized water was added to quench the mixture, and the organic layer was extracted with EA, and the organic layer was washed with saturated NaHCO3 solution and saturated NaCl aqueous solution in turn, and then dried over Na2SO4, concentrated, and column chromatography was performed to obtain the intermediate product 6b. The intermediate product 6b (300 mg) was dissolved in anhydrous DCM (40 mL), and NaHCO3 (330 mg, 4.086 mmol) was added, and then m-CPBA (114.3 mg, 0.663 mmol) was slowly added thereto, and the mixture was stirred at room temperature for 2 h. Subsequently, an appropriate amount of deionized water was added for quenching, the organic layer was extracted with EA, and the organic layer was washed with saturated NaHCO3 solution and saturated NaCl aqueous solution in turn, and then dried over Na2S04 and concentrated. The target compound 8b [(20S, 24R)-epoxy-3,4-opened-3-(3-N-Boc-azetidinamine-1-yl)carbonyl-4(28)-oxirane-dammarane-12β,25-diol] was obtained by column chromatography.

[0078] 1H NMR (400MHz, CDCl3) δ4.59 (qt, J=7.6, 5.4Hz, 1H), 4.25-4.19 (m, 2H), 3.86 (dt, J=8. 8,6.2Hz,1H),3.77-3.71(m,2H),3.51(tt,J=10.9,5.5Hz,1H),2.97-2.88(m,1H),2. 74(t,J=3.7Hz,1H),2.19(ddd,J=11.0,9.1,3.7Hz,1H),2.11-1.46(m,21H),1.44(s ,9H),1.28(s,6H),1.26(s,3H),1.11(s,3H),1.01(s,3H),0.95(s,3H),0.86(s,3H).

[0079] Example 3: (20S, 24R)-epoxy-3,4-opened-3-(4-methylpiperidinol-1-yl)carbonyl-4(28)-oxirane-dammarane-12β,25-diol (Compound 9a);

[0080] Under argon protection, the intermediate product 5 (250 mg, 0.510 mmol) in Example 1 and 4-methylpiperidinol (0.612 mmol) were dissolved in anhydrous DCM (10 mL), EDCI (250 mg, 1.31 mmol) and DMAP (10 mg, 0.08 mmol), stirred at room temperature for 10 min, then an appropriate amount of deionized water was added to quench, the organic layer was extracted with EA, and the organic layer was washed with saturated NaHCO3 solution and saturated NaCl aqueous solution in turn, then dried over Na2SO4, concentrated, and column chromatography to obtain intermediate product 7a. Intermediate product 7a (300 mg) was dissolved in anhydrous DCM (20 mL), and NaHCO3 (330 mg, 4.086 mmol) was added, and then m-CPBA (114.3 mg, 0.663 mmol) was slowly added thereto, and stirred at room temperature for 2 h. Subsequently, an appropriate amount of deionized water was added for quenching, the organic layer was extracted with EA, and the organic layer was washed with saturated NaHCO3 solution and saturated NaCl aqueous solution in turn, and then dried over NaS04 and concentrated. The target compound 9a [(20S, 24R)-epoxy-3,4-opened-3-(4-methylpiperidinol-1-yl)carbonyl-4(28)-oxirane-dammarane-12β,25-diol] was obtained by column chromatography.

[0081] 1H NMR (400MHz, CDCl3) δ4.82(td,J=10.2,5.0Hz,1H),3.86(dd,J=8.6,6.7Hz,1H),3.52(td,J=10.4,4.3Hz,1H),3.30(td,J=12.1,3.0Hz,2H ),2.69(t,J=3.3Hz,1H),2.56-1.28(m,29H),2.01(s,3H),1.27(s,6H),1.26(s,3H),1.10(s,3H),1.01(s,3H),0.97(s,3H),0.88(s,3H).

[0082] Example 4: (20S, 24R)-epoxy-3,4-opened-3-(furfuryl-1-yl)carbonyl-4(28)-oxirane-dammarane-12β,25-diol (Compound 9b);

[0083] Under argon protection, the intermediate product 5 (250 mg, 0.510 mmol) and furfuryl alcohol (0.612 mmol) in Example 1 were dissolved in anhydrous DCM (10 mL), EDCI (250 mg, 1.31 mmol) and DMAP (10 mg, 0.08 mmol) were added, and the mixture was stirred at room temperature for 10 min, and then an appropriate amount of deionized water was added to quench the mixture, and the organic layer was extracted with EA, and the organic layer was washed with a saturated NaHCO3 solution and a saturated NaCl aqueous solution in turn, and then dried over Na2SO4, concentrated, and column chromatography was performed to obtain the intermediate product 7b. The intermediate product 7b (300 mg) was dissolved in anhydrous DCM (20 mL), and NaHCO3 (330 mg, 4.086 mmol) was added, and then m-CPBA (114.3 mg, 0.663 mmol) was slowly added thereto, and the mixture was stirred at room temperature for 2 h. Subsequently, an appropriate amount of deionized water was added for quenching, the organic layer was extracted with EA, and the organic layer was washed with saturated NaHCO3 solution and saturated NaCl aqueous solution in turn, and then dried over NaS04 and concentrated. The target compound 9b [(20S, 24R)-epoxy-3,4-opened-3-(furfuryl-1-yl)carbonyl-4(28)-oxirane-dammarane-12β,25-diol] was obtained by column chromatography.

[0084] 1H NMR(400MHz, CDCl3)δ7.43(d,J=1.9Hz,1H),6.43-6.32(m,2H),5.11-4.98(m,2 H),3.85(dd,J=8.7,6.7Hz,1H),3.49(td,J=10.5,4.4Hz,1H),2.72(d,J=4.2Hz, 1H),2.66(d,J=4.1Hz,1H),2.23-2.13(m,3H),2.11-1.28(m,19H),1.28(s,3H) ,1.26(s,3H),1.26(s,3H),1.10(s,3H),1.00(s,3H),0.96(s,3H),0.84(s,3H).

[0085] Example 5: (20S, 24R)-epoxy-3,4-opened-3-(2-thiazolylmethanol-1-yl)carbonyl-4(28)-oxirane-dammarane-12β,25-diol (Compound 9c);

[0086] Under argon protection, the intermediate product 5 (250 mg, 0.510 mmol) in Example 1 and 2-thiazole methanol (0.612 mmol) were dissolved in anhydrous DCM (10 mL), EDCI (250 mg, 1.31 mmol) and DMAP (10 mg, 0.08 mmol) were added, and the mixture was stirred at room temperature for 10 min, and then an appropriate amount of deionized water was added to quench the mixture, and the organic layer was extracted with EA, and the organic layer was washed with a saturated NaHCO3 solution and a saturated NaCl aqueous solution in turn, and then dried over Na2SO4, concentrated, and column chromatography was performed to obtain the intermediate product 7c. The intermediate product 7c (300 mg) was dissolved in anhydrous DCM (20 mL), and NaHCO3 (330 mg, 4.086 mmol) was added, and then m-CPBA (114.3 mg, 0.663 mmol) was slowly added thereto, and the mixture was stirred at room temperature for 2 h. Subsequently, an appropriate amount of deionized water was added for quenching, the organic layer was extracted with EA, and the organic layer was washed with saturated NaHCO3 solution and saturated NaCl aqueous solution in turn, and then dried over NaS04 and concentrated. The target compound 9c [(20S, 24R)-epoxy-3,4-opened-3-(2-thiazolylmethanol-1-yl)carbonyl-4(28)-oxirane-dammarane-12β,25-diol] was obtained by column chromatography.

[0087] 1H NMR (400MHz, CDCl3) δ7.80 (d, J = 3.2Hz, 1H), 7.41-7.33 (m, 1H), 5.47-5.32 (m, 2H) ,3.86(dd,J=8.7,6.7Hz,1H),3.50(td,J=10.4,4.4Hz,1H),2.76-2.65(m,2H),2.3 1-2.23(m,2H),2.19(ddd,J=11.1,9.2,3.8Hz,1H),2.11-1.12(m,19H),1.28(s,3 H),1.27(s,3H),1.26(s,3H),1.10(s,3H),1.01(s,3H),0.97(s,3H),0.86(s,3H).

[0088] Example 6: (20S, 24R)-epoxy-3,4-opened-3-(2-thiophene methanol-1-yl)carbonyl-4(28)-oxirane-dammarane-12β,25-diol (Compound 9d);

[0089] Under argon protection, the intermediate product 5 (250 mg, 0.510 mmol) in Example 1 and 2-thiophene methanol (0.612 mmol) were dissolved in anhydrous DCM (10 mL), EDCI (250 mg, 1.31 mmol) and DMAP (10 mg, 0.08 mmol) were added, and the mixture was stirred at room temperature for 10 min, and then an appropriate amount of deionized water was added to quench the mixture, and the organic layer was extracted with EA, and the organic layer was washed with a saturated NaHCO3 solution and a saturated NaCl aqueous solution in turn, and then dried over Na2SO4, concentrated, and column chromatography was performed to obtain the intermediate product 7d. The intermediate product 7d (300 mg) was dissolved in anhydrous DCM (20 mL), and NaHCO3 (330 mg, 4.086 mmol) was added, and then m-CPBA (114.3 mg, 0.663 mmol) was slowly added thereto, and the mixture was stirred at room temperature for 2 h. Subsequently, an appropriate amount of deionized water was added for quenching, the organic layer was extracted with EA, and the organic layer was washed with saturated NaHCO3 solution and saturated NaCl aqueous solution in turn, and then dried over NaS04 and concentrated. The target compound 9d [(20S, 24R)-epoxy-3,4-opened-3-(2-thiophenemethylol-1-yl)carbonyl-4(28)-oxirane-dammarane-12β,25-diol] was obtained by column chromatography.

[0090] 1H NMR (400MHz, CDCl3) δ7.32(dd,J=5.1,1.3Hz,1H),7.09(dd,J=3.5,1.1Hz,1H),6.99(dd,J =5.1,3.5Hz,1H),5.32–5.19(m,2H),3.85(dd,J=8.7,6.7Hz,1H),3.49(td,J=10.5,4.5Hz ,1H),2.75-2.69(m,1H),2.65(d,J=4.2Hz,1H),2.23-2.11(m,3H),2.11-1.13(m,19H),1. 28(s,3H),1.26(s,3H),1.25(s,3H),1.10(s,3H),1.00(s,3H),0.96(s,3H),0.84(s,3H).

[0091] Example 7: (20S, 24R)-epoxy-3,4-opened-3-(3-N-Boc-azetidinol-1-yl)carbonyl-4(28)-oxirane-dammarane-12β,25-diol (Compound 9e);

[0092] Under argon protection, the intermediate product 5 (250 mg, 0.510 mmol) in Example 1 and 3-N-Boc-azetidine alcohol (0.612 mmol) were dissolved in anhydrous DCM (10 mL), EDCI (250 mg, 1.31 mmol) and DMAP (10 mg, 0.08 mmol) were added, and the mixture was stirred at room temperature for 10 min, followed by quenching with an appropriate amount of deionized water, and the organic layer was extracted with EA, and the organic layer was washed with a saturated NaHCO3 solution and a saturated NaCl aqueous solution in turn, and then dried over Na2SO4, concentrated, and column chromatography to obtain an intermediate product 7e. The intermediate product 7e (300 mg) was dissolved in anhydrous DCM (20 mL), and NaHCO3 (330 mg, 4.086 mmol) was added, and then m-CPBA (114.3 mg, 0.663 mmol) was slowly added thereto, and stirred at room temperature for 2 h. Subsequently, an appropriate amount of deionized water was added for quenching, the organic layer was extracted with EA, and the organic layer was washed with saturated NaHCO3 solution and saturated NaCl aqueous solution in turn, and then dried over NaS04 and concentrated. The target compound 9e [(20S, 24R)-epoxy-3,4-opened-3-(3-N-Boc-azetidinol-1-yl)carbonyl-4(28)-oxirane-dammarane-12β,25-diol] was obtained by column chromatography.

[0093] 1H NMR (400MHz, CDCl3) δ5.11(tt,J=6.8,4.2Hz,1H),4.23(ddt,J=10.0,6.8,1.1H z,2H),3.92-3.87(m,1H),3.87-3.82(m,2H),3.52(td,J=10.5,4.4Hz,1H),2.76 -2.67(m,2H),2.25-2.15(m,3H),2.12-1.29(m,19H),1.45(s,9H),1.28(s,3H) ,1.27(s,3H),1.27(s,3H),1.10(s,3H),1.02(s,3H),0.97(s,3H),0.87(s,3H).

[0094] Example 8: (20S, 24R)-epoxy-3,4-opened-3-(4-N-Boc-piperidinol-1-yl)carbonyl-4(28)-oxirane-dammarane-12β,25-diol (Compound 9f);

[0095] Under argon protection, the intermediate product 5 (250 mg, 0.510 mmol) in Example 1 and 4-N-Boc-piperidinol (0.612 mmol) were dissolved in anhydrous DCM (10 mL), EDCI (250 mg, 1.31 mmol) and DMAP (10 mg, 0.08 mmol) were added, and the mixture was stirred at room temperature for 10 min, and then an appropriate amount of deionized water was added to quench the mixture, and the organic layer was extracted with EA, and the organic layer was washed with a saturated NaHCO3 solution and a saturated NaCl aqueous solution in turn, and then dried over Na2SO4, concentrated, and column chromatography was performed to obtain the intermediate product 7f. The intermediate product 7f (300 mg) was dissolved in anhydrous DCM (20 mL), and NaHCO3 (330 mg, 4.086 mmol) was added, and then m-CPBA (114.3 mg, 0.663 mmol) was slowly added thereto, and the mixture was stirred at room temperature for 2 h. Subsequently, an appropriate amount of deionized water was added for quenching, the organic layer was extracted with EA, and the organic layer was washed with saturated NaHCO3 solution and saturated NaCl aqueous solution in turn, and then dried over NaS04 and concentrated. The target compound 9f [(20S, 24R)-epoxy-3,4-opened-3-(4-N-Boc-piperidinol-1-yl)carbonyl-4(28)-oxirane-dammarane-12β,25-diol] was obtained by column chromatography.

[0096] 1H NMR(400MHz, CDCl3) δ4.82(tt,J=8.3,3.9Hz,1H),3.83-3.75(m,1H),3.70(d,J =11.5Hz,2H),3.45(td,J=10.5,4.4Hz,1H),3.09(ddd,J=13.1,9.1,3.4Hz,2H) ,2.69(d,J=4.3Hz,1H),2.61(d,J=4.4Hz,1H),2.18-0.96(m,26H),1.40(s,9H) ,1.21(s,3H),1.20(s,6H),1.03(s,3H),0.94(s,3H),0.90(s,3H),0.80(s,3H).

[0097] Example 9: (20S, 24R)-epoxy-3,4-opened-3-(3-oxetanol-1-yl)carbonyl-4(28)-oxirane-dammarane-12β,25-diol (Compound 9g);

[0098] Under argon protection, the intermediate product 5 (250 mg, 0.510 mmol) in Example 1 and 3-oxetane (0.612 mmol) were dissolved in anhydrous DCM (10 mL), EDCI (250 mg, 1.31 mmol) and DMAP (10 mg, 0.08 mmol) were added, and the mixture was stirred at room temperature for 10 min, and then an appropriate amount of deionized water was added to quench the mixture, and the organic layer was extracted with EA, and the organic layer was washed with a saturated NaHCO3 solution and a saturated NaCl aqueous solution in turn, and then dried over Na2SO4, concentrated, and column chromatography was performed to obtain an intermediate product 7g. The intermediate product 7g (300 mg) was dissolved in anhydrous DCM (20 mL), and NaHCO3 (330 mg, 4.086 mmol) was added, and then m-CPBA (114.3 mg, 0.663 mmol) was slowly added thereto, and stirred at room temperature for 2 h. Subsequently, an appropriate amount of deionized water was added for quenching, the organic layer was extracted with EA, and the organic layer was washed with saturated NaHCO3 solution and saturated NaCl aqueous solution in turn, and then dried over NaS04 and concentrated. 9 g of the target compound [(20S, 24R)-epoxy-3,4-opened-3-(3-oxetanol-1-yl)carbonyl-4(28)-oxirane-dammarane-12β,25-diol] was obtained by column chromatography.

[0099] 1H NMR (400MHz, CDCl3) δ5.43(tt,J=6.5,5.3Hz,1H),4.92-4.82(m,2H),4.69-4.59(m,2H),3.86(dd,J=8.7,6.7Hz,1H),3.52(td,J=10.6,4.5Hz, 1H),2.77-2.67(m,2H),2.26-2.15(m,3H),2.13-1.29(m,19H),1.28(s ,3H),1.27(s,6H),1.10(s,3H),1.02(s,3H),0.97(s,3H),0.88(s,3H).

[0100] Example 10: (20S, 24R)-epoxy-3,4-opened-3-(2-pyridinemethanol-1-yl)carbonyl-4(28)-oxirane-dammarane-12β,25-diol (Compound 9h);

[0101] Under argon protection, the intermediate product 5 (250 mg, 0.510 mmol) in Example 1 and 2-pyridine methanol (0.612 mmol) were dissolved in anhydrous DCM (10 mL), EDCI (250 mg, 1.31 mmol) and DMAP (10 mg, 0.08 mmol) were added, and the mixture was stirred at room temperature for 10 min, and then an appropriate amount of deionized water was added to quench the mixture, and the organic layer was extracted with EA, and the organic layer was washed with a saturated NaHCO3 solution and a saturated NaCl aqueous solution in turn, and then dried over Na2SO4, concentrated, and column chromatography was performed to obtain the intermediate product 7h. The intermediate product 7h (300 mg) was dissolved in anhydrous DCM (20 mL), and NaHCO3 (330 mg, 4.086 mmol) was added, and then m-CPBA (114.3 mg, 0.663 mmol) was slowly added thereto, and the mixture was stirred at room temperature for 2 h. Then, an appropriate amount of deionized water was added for quenching, the organic layer was extracted with EA, and the organic layer was washed with saturated NaHCO3 solution and saturated NaCl aqueous solution in turn, and then dried over NaS04 and concentrated. The target compound 9h [(20S, 24R)-epoxy-3,4-opened-3-(2-pyridinemethanol-1-yl)carbonyl-4(28)-oxirane-dammarane-12β,25-diol] was obtained by column chromatography.

[0102] 1H NMR (400MHz, CDCl3) δ8.30(dd,J=6.3,1.3Hz,1H),7.40(d,J=2.2Hz,1H),7.34(ddt,J=7.8,6.1,1.3Hz,1H),5.39(s,2H),3.86(dd,J=8.7,6.7Hz,1H) ,3.53(td,J=10.5,4.4Hz,1H),2.76-2.67(m,2H),2.41-1.28(m,22H),1.2 7(s,6H),1.25(s,3H),1.10(s,3H),1.02(s,3H),0.99(s,3H),0.88(s,3H).

[0103] Example 11: Cytotoxicity Evaluation of PPD Derivatives

[0104] Experimental methods:

[0105] (1) Cell plating: KBV cells in logarithmic growth phase and in good condition were digested and counted, and 2.5×10 3 / well were inoculated in a 96-well plate and cultured in a 37°C, 5% CO2 cell culture incubator;

[0106] (2) Cell administration: After the cells were plated and adhered for 24 hours, 30 μM of different derivatives and corresponding solvents were added for culture, and three parallel wells were set up for each group. After drug addition, the 96-well plate was placed in an incubator and cultured for 72 hours;

[0107] (3) MTT assay: After the cells are cultured with the corresponding drugs for 72 hours, 30 μL of 5 mg / mL MTT solution is added, and the cells are cultured in a 37°C incubator for 2 to 4 hours. The supernatant is discarded, and the crystals formed at the bottom of the wells must not be destroyed. 150 μL DMSO is added to each well, and the cells are placed on a shaker and shaken in the dark for about 10 minutes to fully dissolve the formazan crystals. Finally, the absorbance is detected at a wavelength of 570 nm using an enzyme reader. The tumor cell group treated with DMSO was used as the control group, and verapamil was used as the positive control to calculate the inhibition rate of the derivative. Inhibition rate (%) = (average OD value of the control group – average OD value of the drug-treated group) / average value of the control group × 100%

[0108] Experimental results: When the drug concentration was 30 μM, the survival rate of the tumor-resistant KBV cells treated with Vrp was about 40%, while the survival rates of the tumor-resistant KBV cells treated with compounds 8a-8b and 9a-9h were all above 80%, indicating that the cytotoxicity of these compounds was significantly lower than that of Vrp.

[0109] Table 1 Evaluation results of cytotoxicity of PPD derivatives to KBV cells

[0110]

[0111] Example 12: Evaluation of the anti-tumor MDR activity of PPD derivatives

[0112] Experimental methods:

[0113] (1) Cell plating: KBV cells in logarithmic growth phase and in good condition were digested and counted, and 2.5×10 3 / well were inoculated in a 96-well plate and cultured in a 37°C, 5% CO2 cell culture incubator;

[0114] (2) Cell administration: After the cells were plated and adhered for 24 hours, 10 μM of different derivatives were added and combined with 100 nM paclitaxel and DMSO as control cultures. Three parallel wells were set up for each group. After drug addition, the 96-well plate was placed in an incubator and cultured for 72 hours;

[0115] (3) MTT assay: After the cells are cultured with the corresponding drugs for 72 hours, 30 μL of 5 mg / mL MTT solution is added, and the cells are cultured in a 37°C incubator for 2 to 4 hours. The supernatant is discarded, and the crystals formed at the bottom of the wells must not be destroyed. 150 μL DMSO is added to each well, and the cells are placed on a shaker and shaken in the dark for about 10 minutes to fully dissolve the formazan crystals. Finally, the absorbance is detected at a wavelength of 570 nm using an enzyme reader. The tumor cell group treated with DMSO was used as the control group, and verapamil was used as the positive control to calculate the inhibition rate of the derivative. Inhibition rate (%) = (average OD value of the control group – average OD value of the drug-treated group) / average value of the control group × 100%

[0116] Experimental results: Paclitaxel has no cytotoxicity to the tumor-resistant cell line KBV at 100nM. When PPD derivatives at a concentration of 10μM are added, the survival rate of KBV cells is significantly reduced, indicating that the sensitivity of paclitaxel-resistant tumor-resistant cells KBV to paclitaxel is significantly improved, that is, it shows a tumor resistance reversal effect. Compared with PPD derivatives 6a-6b and 7a-7h without covalent cross-linking groups, PPD derivatives 8a-8b and 9a-9h with covalent cross-linking groups formed by epoxy ring formation have better tumor resistance reversal activity. Among them, the tumor resistance reversal activity of preferred compounds 8a, 8b, 9c, 9d, 9e, and 9f is significantly better than that of the positive drug verapamil (Vrp).

[0117] Table 2 Evaluation results of the antitumor MDR activity of PPD derivatives on KBV cells

[0118]

[0119]

[0120] The above description is only a partial embodiment of the present application and an illustration of the technical principles used.

[0121] Those skilled in the art should understand that the scope of the invention involved in this application is not limited to the technical solutions formed by the combination of the above technical features, but should also cover other technical solutions formed by any combination of the above technical features or their equivalent features without departing from the inventive concept. For example, the above features are replaced with but not limited to the technical features with similar functions disclosed in this application.

Claims

1. A PPD ring-opening epoxy substituted derivative, whose structural formula is shown in Formula 8 or Formula 9, or a pharmaceutically acceptable salt of the compound shown in Formula 8 or Formula 9, in, R1 and R2 are each independently R x -K-, where K represents a methylene group or does not exist, R x It is a C3-C10 heterocyclic group or heteroaromatic group which is unsubstituted or substituted with one or more substituents; the substituent is a C1-C10 alkyl group or tert-butyloxycarbonyl group.

2. The PPD ring-opening epoxy substituted derivative according to claim 1, characterized in that R x It is one of pyrazine, pyridine, azetidine, thiophene, piperidine, furan, tetrahydrofuran, and thiazole.

3. The PPD ring-opening epoxy substituted derivative according to claim 1, characterized in that R1 represents 1-methyl-(2-thiophene)-1-yl or 3-N-Boc-azetidine-1-yl; R2 represents 4-methylpiperidin-1-yl, 1-methyl-(2-furan)-1-yl, 1-methyl-(2-thiazole)-1-yl, 1-methyl-(2-thiophene)-1-yl, 3-N-Boc-azetidine-1-yl, 4-N-Boc-piperidin-1-yl, 3-oxetan-1-yl, or 1-methyl-(2-pyridine)-1-yl.

4. The PPD ring-opening epoxy substituted derivative according to claim 1, characterized in that The PPD ring-opening epoxy substituted derivative is one of the following compounds:

5. The method for preparing the ring-opening epoxy substituted derivative of PPD according to claim 1, characterized in that: The method is shown in the following reaction formula:

6. Use of the PPD ring-opened epoxy substituted derivative and a pharmaceutically acceptable salt thereof as claimed in any one of claims 1 to 4 in the preparation of a tumor resistance reversal agent / tumor drug sensitizer, or in the preparation of a tumor resistance reversal agent / tumor drug sensitizer containing a pharmaceutically acceptable carrier.

7. The use according to claim 6, characterized in that The PPD ring-opening epoxy substituted derivative and its pharmaceutically acceptable salt are used in preparing a tumor resistance reversal agent / tumor drug sensitizer that overcomes P-glycoprotein mediation or in preparing a tumor resistance reversal agent / tumor drug sensitizer that contains a pharmaceutically acceptable carrier.

8. Use of the PPD ring-opened epoxy substituted derivative and its pharmaceutically acceptable salt as described in any one of claims 1 to 4 in the preparation of anti-tumor drugs, wherein in the method of application, the anti-tumor drug includes clinical anti-tumor drugs and tumor resistance reversal agents / tumor drug sensitizers, and the tumor resistance reversal agents / tumor drug sensitizers are PPD ring-opened epoxy substituted derivatives and their pharmaceutically acceptable salts as shown in Formula 8 or Formula 9.

9. The use according to claim 8, characterized in that The tumor is breast cancer, colon cancer, cervical cancer, liver cancer, stomach cancer or lung cancer; The clinical anti-tumor drug is paclitaxel.

10. A pharmaceutical composition for combating multidrug resistance, comprising the ring-opening epoxy-substituted derivative of PPD according to any one of claims 1 to 4 and a pharmaceutically acceptable salt thereof.

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