PPD ring-opened derivative as well as preparation method and application thereof

Abstract

The invention discloses a PPD ring-opened derivative, which has a structural formula as shown in a formula 11, 12, 13 or 14, or a pharmaceutically acceptable salt of a compound as shown in the formula 11, 12, 13 or 14. The invention discloses a PPD ring-opening derivative, a preparation method of the PPD ring-opening derivative, and application of the PPD ring-opening 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 derivative disclosed by the invention has lower cytotoxicity and better tumor drug resistance reversal effect, and the effect is obviously superior to that of a positive control drug. The PPD ring-opening derivative disclosed by the invention has relatively high development and application values. # 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 derivative, a preparation method thereof and an application thereof in the field of medicine. Background Art

[0002] Malignant tumors seriously affect human health. Chemotherapy is one of the effective means to treat malignant tumors, but the occurrence of tumor multidrug resistance (MDR) will greatly reduce the efficacy of drugs, adding great difficulties to tumor treatment. Overexpression of P-glycoprotein (P-gp) is one of the important reasons for tumor multidrug resistance. P-gp inhibitors are expected to become key drugs to overcome tumor multidrug resistance. However, existing P-gp inhibitors have problems such as low efficacy and high toxicity. Therefore, new structural types of P-gp inhibitors need to be sought.

[0003] Ginseng is a commonly used Chinese herbal medicine. Ginsenosides are active chemical components extracted from ginseng, which have multiple pharmacological activities such as antioxidant, anti-inflammatory, antibacterial, anti-cancer and anti-myocardial ischemia. Among them, 20S-protopanaxadiol (PPD) is the main sapogenin and is now commonly used as an auxiliary drug during the treatment of cancer patients. 20S-PPD is easily metabolized into a furan ring structure 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 formulas A, B, C, and D, 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 derivative, whose structural formula is shown in Formula 11, 12, 13 or 14, or a pharmaceutically acceptable salt of the compound shown in Formula 11, 12, 13 or 14,

[0007]

[0008] Among them, R 1 , R 2 , R 3 , R 4 Each is independent of R x-KY-, where Y represents NH or O, K represents methylene or is absent, R x is a C3-C10 heterocyclic or heteroaromatic group which is unsubstituted or substituted with one or more substituents. A heterocyclic or heteroaromatic group refers to a heterocyclic or heteroaromatic group containing one or both of N and O atoms in the ring. x The substituents on may be C1-C10 alkyl or tert-butyloxycarbonyl;

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

[0010] Furthermore, R 1 represents 2-pyrazinemethanamine-1-yl, 4-pyridinemethanol-1-yl, 3-N-Boc-azetidinol-1-yl, 3-azetidinol-1-yl, 2-thiophenemethanamine-1-yl, 3-N-Boc-azetidinamine-1-yl or 4-N-Boc-piperidinol-1-yl; R 2 represents 4-pyridinemethanol-1-yl or 3-tetrahydrofuranol-1-yl; R 3 represents 2-thiazolylmethanol-1-yl, 2-pyridinemethanol-1-yl, 4-pyridinemethanol-1-yl or 2-pyrazinemethanol-1-yl; R 4 represents furfuryl-1-yl, 2-thiazolylmethanol-1-yl, 2-pyridiniummethanol-1-yl, 4-pyridiniummethanol-1-yl or 2-pyraziniummethanol-1-yl.

[0011] The PDD ring-opening derivative represented by Formula 11, 12, 13 or 14 of the present invention is preferably one of the following:

[0012] (20S,24R)-epoxy-3,4-opened-3-(2-pyrazinemethylamine-1-yl)carbonyl-4(28)-ene-dammarane-12β,25-diol (Compound 11a);

[0013] (20S,24R)-epoxy-3,4-opened-3-(4-pyridylmethanol-1-yl)carbonyl-4(28)-ene-dammarane-12β,25-diol (Compound 11b);

[0014] (20S,24R)-epoxy-3,4-opened-3-(3-N-Boc-azetidinol-1-yl)carbonyl-4(28)-ene-dammarane-12β,25-diol (Compound 11c);

[0015] (20S,24R)-epoxy-3,4-seco-3-(3-azetidinol-1-yl)carbonyl-4(28)-ene-dammarane-12β,25-diol (Compound 11d);

[0016] (20S,24R)-epoxy-3,4-opened-3-(3-N-Boc-azetidinamine-1-yl)carbonyl-4(28)-ene-dammarane-12β,25-diol (Compound 11f);

[0017] (20S,24R)-Epoxy-3,4-opened-3-(4-N-Boc-piperidinol-1-yl)carbonyl-4(28)-ene-dammarane-12β,25-diol (Compound 11g);

[0018] (20S,24S)-epoxy-3,4-opened-3-(4-pyridylmethanol-1-yl)carbonyl-4(28)-ene-dammarane-12β,25-diol (Compound 12a);

[0019] (20S,24S)-epoxy-3,4-opened-3-(3-tetrahydrofuranol-1-yl)carbonyl-4(28)-ene-dammarane-12β,25-diol (Compound 12b);

[0020] (20S,24R)-epoxy-3,4-opened-3-(2-thiazolylmethanol-1-yl)carbonyl-dammarane-4,12β,25-triol (Compound 13a);

[0021] (20S,24R)-epoxy-3,4-opened-3-(2-pyridylmethanol-1-yl)carbonyl-dammarane-4,12β,25-triol (Compound 13b);

[0022] (20S,24R)-epoxy-3,4-opened-3-(4-pyridylmethanol-1-yl)carbonyl-dammarane-4,12β,25-triol (Compound 13c);

[0023] (20S,24R)-epoxy-3,4-opened-3-(4-pyrazinemethanol-1-yl)carbonyl-dammarane-4,12β,25-triol (Compound 13d);

[0024] (20S,24S)-epoxy-3,4-opened-3-(furfuryl-1-yl)carbonyl-dammarane-4,12β,25-triol (Compound 14a);

[0025] (20S,24S)-epoxy-3,4-opened-3-(2-thiazolylmethanol-1-yl)carbonyl-dammarane-4,12β,25-triol (Compound 14b);

[0026] (20S,24S)-epoxy-3,4-opened-3-(2-pyridylmethanol-1-yl)carbonyl-dammarane-4,12β,25-triol (Compound 14c);

[0027] (20S,24S)-epoxy-3,4-opened-3-(4-pyridylmethanol-1-yl)carbonyl-dammarane-4,12β,25-triol (Compound 14d);

[0028] (20S,24S)-Epoxy-3,4-opened-3-(2-pyrazinemethanol-1-yl)carbonyl-dammarane-4,12β,25-triol (Compound 14e).

[0029] The structural formulas of compounds 11a-11g, 12a-12b, 13a-13d, and 14a-14e are shown below:

[0030]

[0031]

[0032] More preferably, the PPD ring-opening derivative is a compound represented by Formula 11a, 11b, 11c, 11d, 11f, 11g, 12a, 12b, 13a, 13b, 13c, 14a or 14d.

[0033] The present invention also provides the use of the PPD open-ring derivatives shown in Formula 11, 12, 13 or 14 and pharmaceutically acceptable salts thereof in the preparation of tumor resistance reversal agents / tumor drug sensitizers, or in the preparation of tumor resistance reversal agents / tumor drug sensitizers containing pharmaceutically acceptable carriers; further, the PPD open-ring derivatives shown in Formula 11, 12, 13 or 14 and pharmaceutically acceptable salts thereof can be used to prepare tumor resistance reversal agents / tumor drug sensitizers that overcome P-glycoprotein-mediated tumor resistance or to prepare tumor resistance reversal agents / tumor drug sensitizers containing pharmaceutically acceptable carriers.

[0034] Furthermore, in the application, the PPD open-ring derivative is preferably a compound represented by Formula 11a, 11b, 11c, 11d, 11f, 11g, 12a, 12b, 13a, 13b, 13c, 14a or 14d.

[0035] The present invention also provides the use of PPD open-ring derivatives shown in Formula 11, 12, 13 or 14 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 open-ring derivatives shown in Formula 11, 12, 13 or 14 and pharmaceutically acceptable salts thereof.

[0036] Furthermore, the tumor is breast cancer, colon cancer, cervical cancer, liver cancer, gastric cancer or lung cancer.

[0037] The clinical anti-tumor drug is paclitaxel.

[0038] Furthermore, in the application, the PPD open-ring derivative is preferably a compound represented by Formula 11a, 11b, 11c, 11d, 11f, 11g, 12a, 12b, 13a, 13b, 13c, 14a or 14d.

[0039] The present invention also provides the use of the PPD open-ring derivatives shown in Formula 11, 12, 13 or 14 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 multidrug-resistant tumor diseases.

[0040] 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.

[0041] The present invention also provides a pharmaceutical composition for resisting multidrug resistance, which comprises a PPD open-ring derivative shown in Formula 11, 12, 13 or 14 and a pharmaceutically acceptable salt thereof, and can also be added with a pharmaceutically acceptable carrier to prepare a pharmaceutical preparation such as a tablet, capsule, powder, syrup, suspension, injection, etc.; and can also be added with commonly used pharmaceutical excipients such as sweeteners, diluents, fillers, etc.

[0042] Furthermore, in the pharmaceutical composition, the PPD open-ring derivative is preferably of Chemical Formula 11a, 11b, 11c, 11d, 11f, 11g, 12a, 12b, 13a, 13b, 13c, 14a or 14d.

[0043] The pharmaceutical composition provided by the present invention can be administered clinically by oral administration, injection or the like.

[0044] 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.

[0045] Furthermore, the present invention also provides a method for preparing the PPD open-ring derivative shown in Formula 11, 12, 13 or 14, and the method is shown in the following reaction formula:

[0046]

[0047] The method comprises the following steps:

[0048] (1) 24R-Pyxinol represented by Formula 1 or 24S-Pyxinol represented by Formula 2 is added with pyridinium chlorochromate (PCC) at a molar ratio of 1:0.5 to 4 to carry out an oxidation reaction, thereby obtaining Compound 3 or Compound 4, respectively;

[0049] The reaction solvent of step (1) is an aprotic solvent, preferably dichloromethane (DCM) or tetrahydrofuran;

[0050] The reaction temperature of the oxidation reaction is -20°C to 80°C; preferably, the reaction temperature is -10°C to 30°C, and more preferably, it is room temperature.

[0051] 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.

[0052] After the reaction is completed, the reaction solution is post-treated to obtain compound 3 or compound 4. 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 or compound 4.

[0053] (2) Compound 3 or Compound 4 is reacted with m-chloroperbenzoic acid to undergo Baeyer-Villiger reaction to obtain Compound 5 or Compound 6;

[0054] The molar ratio of compound 3 or compound 4 to meta-chloroperbenzoic acid (m-CPBA) is 1:3-15.

[0055] Further, the reaction solvent of step (2) is an aprotic solvent, preferably dichloromethane (DCM) or tetrahydrofuran;

[0056] The reaction temperature of the Baeyer-Villiger reaction is 10° C. to 40° C., preferably room temperature; and the reaction time is 5 to 12 hours.

[0057] After the reaction is completed, the reaction solution is post-treated to obtain compound 5 or compound 6. The post-treatment method of the reaction solution is: the reaction solution is stirred, filtered, vacuum concentrated, and then purified by silica gel column chromatography (SGCC) to obtain compound 5 or compound 6 respectively.

[0058] (3) Compound 5 or Compound 6 is subjected to an acidic ring-opening reaction with p-toluenesulfonamide (P-TSA) to obtain Compound 7 or Compound 8, respectively;

[0059] The molar ratio of the compound 5 or compound 6 to p-toluenesulfonamide (P-TSA) is 1:3-9.

[0060] Further, the reaction solvent of step (3) is an aprotic solvent, preferably dichloromethane (DCM) or tetrahydrofuran;

[0061] The reaction temperature of the acidic ring-opening reaction is -10°C to 60°C, preferably 10°C to 40°C, more preferably room temperature; the reaction time is 5 to 18 hours.

[0062] After the reaction is completed, the reaction solution is post-treated to obtain compound 7 or compound 8. The post-treatment method of the reaction solution is: the reaction solution is stirred, filtered, vacuum concentrated, and then purified by silica gel column chromatography (SGCC) to obtain compound 7 or compound 8 respectively.

[0063] (4) Compound 5 or Compound 6 is subjected to an alkaline ring-opening reaction with potassium hydroxide (KOH) to obtain Compound 9 or Compound 10, respectively;

[0064] The molar ratio of the compound 5 or compound 6 to potassium hydroxide is 1:2-10.

[0065] Furthermore, the reaction solvent in step (4) is an aprotic solvent, preferably a tetrahydrofuran aqueous solution (THF / H 2 O); the volume ratio of tetrahydrofuran to water in the tetrahydrofuran aqueous solution is preferably 10 to 20:1.

[0066] The reaction temperature of the alkaline ring-opening reaction is -10°C to 40°C, preferably 10°C to 40°C, more preferably room temperature; the reaction time is 1 to 2 days, preferably 5 to 18 hours.

[0067] After the reaction is completed, the reaction solution is post-treated to obtain compound 9 or compound 10. The post-treatment method of the reaction solution is: evaporating the solvent of the reaction solution under vacuum, and purifying the concentrated sample by silica gel column chromatography to obtain compound 9 or compound 10 respectively.

[0068] (5) Compound 7, Compound 8, Compound 9 or Compound 10 are respectively mixed with an amino-containing heterocyclic compound (R 1 -NH 2 , R 2 -NH 2 , R 3 -NH 2 , R 4 -NH 2 ) or hydroxyl-containing heterocyclic compounds (R 1 -OH, R 2 -OH, R 3 -OH, R 4 -OH) to undergo amide reaction or esterification reaction to obtain the PPD ring-opening derivatives shown in formula 11, 12, 13 or 14, respectively.

[0069] The molar ratio of compound 7, compound 8, compound 9 or compound 10 to the amino-containing heterocyclic compound or the hydroxyl-containing heterocyclic compound is 1:1.2-10;

[0070] Furthermore, the catalyst for the amide reaction is HBTU (benzotriazole-N,N,N,N-tetramethyluronium hexafluorophosphate) and triethylamine, and the molar ratio of compound 7, compound 8, compound 9 or compound 10 to HBTU and triethylamine is 1:1.3-5:2-10.

[0071] Furthermore, the catalysts for the esterification reaction are EDCI and DMAP, and the molar ratio of compound 7, compound 8, compound 9 or compound 10 to EDCI and DMAP is 1:2-5:0.1-1.

[0072] Further, the reaction solvent of step (5) is an aprotic solvent, preferably N,N-dimethylformamide or dichloromethane (DCM);

[0073] The reaction temperature of the amide reaction or the esterification reaction is -10°C to 40°C, preferably 5°C to 40°C, more preferably room temperature; the reaction time is 0.5 to 16 hours.

[0074] After the reaction is completed, the resulting reaction solution is post-treated to obtain the PPD open-ring derivatives shown in Formula 11, 12, 13 or 14, 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 the PPD open-ring derivatives shown in Formula 11, 12, 13 or 14, respectively.

[0075] Compared with the prior art, the present invention has the following advantages:

[0076] 1. The PPD ring-opening derivatives represented by formulas (11a-11g), (12a-12b), (13a-13d), and (14a-14e) of the present invention are synthesized for the first time, and compared with their respective parent core compounds 7-10, their anti-tumor MDR activity is discovered for the first time.

[0077] 2. The PPD derivatives represented by formula (11a-11g), (12a-12b), (13a-13d), and (14a-14e) 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 11a-11g, 12a-12b, 13a-13d, and 14a-14e do not show cytotoxicity, and the survival rate is above 80%.

[0078] 3. The compounds of the present invention have better tumor resistance reversal effect. The PPD derivatives shown in formulas (11a-11g), (12a-12b), (13a-13d), and (14a-14e) of the present invention have better tumor resistance reversal ability and activity in inhibiting P-glycoprotein function compared with sapogenin PPD and the parent compound 7-10. For example, the commonly used clinical drug paclitaxel has no cytotoxicity to the tumor-resistant cell line KBV at 100nM. When sapogenin PPD and the parent compound 7-10 were added in a concentration of 10μM, no cytotoxicity was shown, indicating that at this concentration, these compounds failed to increase the sensitivity of paclitaxel-resistant tumor-resistant cells KBV to paclitaxel. When compounds 11a, 11b, 11c, 11d, 11f, 11g, 12a, 12b, 13a, 13b, 13c, 14a and 14d were added together, the survival rate of KBV cells was significantly reduced, indicating that the sensitivity of KBV cells to paclitaxel was significantly improved, that is, it showed a tumor resistance reversal effect, and the effect was significantly better than the positive control drug verapamil (Vrp). The PPD open-ring derivatives of the present invention have strong development and application value. DETAILED DESCRIPTION

[0079] 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.

[0080] Example 1: (20S, 24R)-epoxy-3,4-opened-3-(2-pyrazinemethylamine-1-yl)carbonyl-4(28)-ene-dammarane-12β,25-diol (11a);

[0081] Protopanaxadiol (20S-PPD) (100 mg, 0.217 mmol) was used as the raw material and dissolved in anhydrous dichloromethane (2.17 mL). The oxidative cyclization reaction was carried out using m-CPBA (54.933 mg, 0.318 mmol). After stirring at room temperature for 12 h, the mixture was filtered and concentrated in vacuo. Silica gel column chromatography (SGCC) was then used to obtain the intermediates 1 and 2. The intermediate product 1 (821 mg, 1.72 mmol) was dissolved in anhydrous dichloromethane (20 mL), and pyridinium chlorochromate (389 mg, 1.80 mmol) was added. After stirring at room temperature for 12 h, the mixture was filtered and concentrated in vacuo. Then, the intermediate product 3 was obtained by silica gel column chromatography (SGCC). Compound 3 (26 mg, 0.054 mmol) was dissolved in 0.5 mL of dichloromethane, and m-CPBA (101 mg, 0.438 mmol) was added. After stirring at room temperature overnight, the mixture was filtered and concentrated in vacuo. Then, the intermediate product 5 was obtained by silica gel column chromatography (SGCC). Compound 5 (22 mg, 0.045 mmol) was dissolved in anhydrous dichloromethane (0.9 mL), and P-TSA (68.3 mg, 0.359 mmol) was added. After stirring at room temperature for 12 h, the mixture was filtered and concentrated in vacuo. Silica gel column chromatography (SGCC) was used to obtain intermediate 7. Compound 7 (250 mg, 0.510 mmol) and 2-aminomethylpyrazine (0.612 mmol) were dissolved in anhydrous DCM (10 mL), and HBTU (240 mg, 0.76 mmol) and Et 3 N (200 μL), stirred at room temperature for about 5 minutes, then added an appropriate amount of deionized water to quench, extracted the organic layer with EA, and then washed with saturated NaHCO 3 The organic layer was washed with saturated NaCl solution and then with Na 2 SO 4 The residue was dried, concentrated and purified by column chromatography to obtain the target compound 11a [(20S,24R)-epoxy-3,4-oxy-3-(2-pyrazinemethylamine-1-yl)carbonyl-4(28)-ene-dammarane-12β,25-diol].

[0082] 1 H NMR (400 MHz, CDCl 3)δ8.61(d,J=1.4Hz,1H),8.56–8.48(m,2H),4.87(t,J=1.8Hz,1H),4.71(d,J=2.2Hz,1H) ,4.62(dd,J=16.8,5.1Hz,1H),4.56(dd,J=16.7,5.0Hz,1H),3.86(dd,J=8.7,6.6Hz,1H) ,3.54(td,J=10.5,4.4Hz,1H),2.41–2.29(m,1H),2.25–2.10(m,2H),2.09–1.13(m,19H) ,1.76(d,J=1.2Hz,3H),1.27(s,6H),1.10(s,3H),1.03(s,3H),0.92(s,3H),0.87(s,3H).

[0083] Example 2: (20S, 24R)-epoxy-3,4-opening-3-(4-pyridinemethanol-1-yl)carbonyl-4(28)-ene-dammarane-12β,25-diol (11b);

[0084] Under argon protection, compound 7 (250 mg, 0.510 mmol) in Example 1 and 4-pyridinemethanol (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 at 0 ° C. The mixture was stirred at room temperature for about 30 minutes, and then an appropriate amount of deionized water was added for quenching. The organic layer was extracted with EA, and then saturated NaHCO 3 The organic layer was washed with saturated NaCl solution and then with Na 2 SO 4 The reaction mixture was dried and concentrated, and the concentrated sample was purified by silica gel column chromatography. According to the adsorption properties of the target product on silica gel, the target compound 11b [(20S, 24R)-epoxy-3,4-opened-3-(4-pyridinemethanol-1-yl)carbonyl-4(28)-ene-dammarane-12β,25-diol] was obtained by column chromatography.

[0085] 1 H NMR (400 MHz, CDCl 3)δ8.64–8.58(m,2H),7.27–7.22(m,2H),5.10(s,2H),4.85(t,J=1.8Hz,1H),4.67(d,J=2.2Hz, 1H), 3.86 (dd, J=8.7, 6.5Hz, 1H), 3.54 (td, J=10.5, 4.5Hz, 1H), 2.43 (ddd, J=15.8, 10.1, 6.1Hz, 1H),2.29(ddd,J=15.1,11.2,7.2Hz,1H),2.20(ddd,J=10.9,9.0,3.7Hz,1H),2.11–0.83(m,19 H),1.73(s,3H),1.28(s,3H),1.27(s,3H),1.10(s,3H),1.03(s,3H),0.92(s,3H),0.87(s,3H).

[0086] Example 3: (20S, 24R)-epoxy-3,4-opened-3-(3-N-Boc-azetidinol-1-yl)carbonyl-4(28)-ene-dammarane-12β,25-diol (11c);

[0087] Under argon protection, the intermediate product 7 in Example 1 (250 mg, 0.510 mmol) and N-Boc-3-hydroxyazetidine (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 at 0 ° C. The mixture was stirred at room temperature for about 30 minutes, and then an appropriate amount of deionized water was added for quenching. The organic layer was extracted with EA, and then saturated NaHCO 3 The organic layer was washed with saturated NaCl solution and then with Na 2 SO 4 The reaction mixture was dried and concentrated, and the concentrated sample was purified by silica gel column chromatography. According to the adsorption properties of the target product on silica gel, the target compound 11c [(20S,24R)-epoxy-3,4-opened-3-(3-N-Boc-azetidinol-1-yl)carbonyl-4(28)-ene-dammarane-12β,25-diol] was obtained by column chromatography.

[0088] 1 H NMR (400 MHz, CDCl 3)δ5.10(tt,J=6.8,4.3Hz,1H),4.86(t,J=1.8Hz,1H),4.66(d,J=1.4Hz,1H),4.21(ddt,J=10. 0,6.8,1.2Hz,2H),3.90–3.87(m,1H),3.87–3.82(m,2H),3.53(td,J=10.6,4.5Hz,1H),2.36( ddd,J=15.7,10.3,6.0Hz,1H),2.26–2.14(m,2H),2.11–0.83(m,19H),1.73(d,J=0.4Hz,3H), ,1.45(s,9H),,1.28(s,3H),1.28(s,3H),1.10(s,3H),1.03(s,3H),0.92(s,3H),0.86(s,3H).

[0089] Example 4: (20S, 24R)-epoxy-3,4-opened-3-(3-azetidinol-1-yl)carbonyl-4(28)-ene-dammarane-12β,25-diol (11d);

[0090] Under argon protection, the intermediate product 7 in Example 1 (250 mg, 0.510 mmol) and azetidine-3-ol (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 at 0 ° C. The mixture was stirred at room temperature for about 30 minutes, and then an appropriate amount of deionized water was added for quenching. The organic layer was extracted with EA, and then saturated NaHCO 3 The organic layer was washed with saturated NaCl solution and then with Na 2 SO 4 The reaction mixture was dried and concentrated, and the concentrated sample was purified by silica gel column chromatography. According to the adsorption properties of the target product on silica gel, the target compound 11d [(20S, 24R)-epoxy-3,4-opened-3-(3-azetidinol-1-yl)carbonyl-4(28)-ene-dammarane-12β,25-diol] was obtained by column chromatography.

[0091] 1 H NMR (400 MHz, CDCl 3)δ5.30(s,1H),4.85(t,J=1.8Hz,1H),4.65(d,J=2.1Hz,1H),4.55–4.02( m,4H),3.86(t,J=7.7Hz,1H),3.56(t,J=7.9Hz,1H),2.43–2.34(m,1H),2. 31–2.17(m,2H),2.09–1.79(m,8H),1.72(s,3H),1.71–0.88(m,11H),1.28 (s,3H),1.27(s,3H),1.12(s,3H),1.03(s,3H),0.93(s,3H),0.87(s,3H).

[0092] Example 5: (20S, 24R)-epoxy-3,4-opened-3-(2-thiophenemethylamine-1-yl)carbonyl-4(28)-ene-dammarane-12β,25-diol (11e);

[0093] Under argon protection, the intermediate product 7 in Example 1 (250 mg, 0.510 mmol) and 2-thiophenemethylamine (0.612 mmol) were dissolved in anhydrous DCM (10 mL), and HBTU (240 mg, 0.76 mmol) and Et 3 N (200 μL), stirred at room temperature for about 5 minutes, then added an appropriate amount of deionized water to quench, extracted the organic layer with EA, and then washed with saturated NaHCO 3 The organic layer was washed with saturated NaCl solution and then with Na 2 SO 4 The residue was dried, concentrated and purified by column chromatography to obtain the target compound 11e [(20S,24R)-epoxy-3,4-oxy-3-(2-thiophenemethylamine-1-yl)carbonyl-4(28)-ene-dammarane-12β,25-diol].

[0094] 1 H NMR (400 MHz, CDCl 3)δ7.22(dd,J=4.9,1.5Hz,1H),6.99–6.91(m,2H),4.87(t,J=1.8Hz,1H),4.70(d,J=2.1 Hz,1H),4.61(dd,J=15.3,5.7Hz,1H),4.55(dd,J=15.2,5.5Hz,1H),3.85(dd,J=8.7,6.5 Hz,1H),3.52(td,J=10.5,4.4Hz,1H),2.36–2.24(m,1H),2.24–2.16(m,1H),2.11–1.12 (m,20H),1.75(s,3H),1.27(s,6H),1.10(s,3H),1.02(s,3H),0.90(s,3H),0.86(s,3H).

[0095] Example 6: (20S, 24R)-epoxy-3,4-opened-3-(3-N-Boc-azetidinamine-1-yl)carbonyl-4(28)-ene-dammarane-12β,25-diol (11f);

[0096] Under argon protection, the intermediate product 7 in Example 1 (250 mg, 0.510 mmol) and N-Boc-3-hydroxyazetidine (0.612 mmol) were dissolved in anhydrous DCM (10 mL), and HBTU (240 mg, 0.76 mmol) and Et 3 N (200 μL), stirred at room temperature for about 5 minutes, then added an appropriate amount of deionized water to quench, extracted the organic layer with EA, and then washed with saturated NaHCO 3 The organic layer was washed with saturated NaCl solution and then with Na 2 SO 4 The residue was dried, concentrated and purified by column chromatography to obtain the target compound 11f [(20S,24R)-epoxy-3,4-oxygen-3-(3-N-Boc-azetidinamine-1-yl)carbonyl-4(28)-ene-dammarane-12β,25-diol].

[0097] 1 H NMR (400 MHz, CDCl 3)δ4.88(t,J=1.5Hz,1H),4.70(d,J=2.2Hz,1H),4.64–4.50(m,1H),4.27(d t,J=37.5,8.5Hz,2H),3.86(dd,J=8.7,6.6Hz,1H),3.72(dd,J=9.3,5.3Hz ,2H),3.55(td,J=10.5,4.4Hz,1H),2.32–1.13(m,22H),1.76(s,3H),1.44 (s,9H),1.28(s,6H),1.11(s,3H),1.02(s,3H),0.91(s,3H),0.86(s,3H).

[0098] Example 7: (20S, 24R)-epoxy-3,4-opening-3-(4-N-Boc-piperidinol-1-yl)carbonyl-4(28)-ene-dammarane-12β,25-diol (11 g);

[0099] Under argon protection, the intermediate product 7 in Example 1 (250 mg, 0.510 mmol) and N-Boc-3-hydroxypiperidine (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 at 0 ° C. The mixture was stirred at room temperature for about 30 minutes, and then an appropriate amount of deionized water was added for quenching. The organic layer was extracted with EA, and then saturated NaHCO 3 The organic layer was washed with saturated NaCl solution and then with Na 2 SO 4 The mixture was dried and concentrated, and the concentrated sample was purified by silica gel column chromatography. According to the adsorption properties of the target product on silica gel, 11 g of the target compound [(20S, 24R)-epoxy-3,4-opened-3-(4-N-Boc-piperidinol-1-yl)carbonyl-4(28)-ene-dammarane-12β,25-diol] was obtained by column chromatography.

[0100] 1 H NMR (400 MHz, CDCl 3)δ4.94–4.83(m,2H),4.68–4.65(m,1H),3.86(dd,J=8.7,6.6Hz,1H),3.71(dd ,J=13.3,6.5Hz,2H),3.54(td,J=10.6,4.4Hz,1H),3.25–3.13(m,2H),2.33(d dd,J=15.6,10.4,5.6Hz,1H),2.24–1.30(m,25H),1.73(s,3H),1.46(s,9H),1 .28(s,3H),1.27(s,3H),1.10(s,3H),1.02(s,3H),0.92(s,3H),0.85(s,3H).

[0101] Example 8: (20S, 24S)-epoxy-3,4-opened-3-(4-pyridylmethanol-1-yl)carbonyl-4(28)-ene-dammarane-12β,25-diol (12a);

[0102] Protopanaxadiol (20S-PPD) (100 mg, 0.217 mmol) was used as the raw material, dissolved in anhydrous dichloromethane (2.17 mL), and oxidative cyclization reaction was carried out using m-CPBA (54.933 mg, 0.318 mmol). After stirring at room temperature for 12 h, the mixture was filtered and concentrated in vacuo, and then 24S-Pyxinol was obtained by silica gel column chromatography (SGCC). 24S-Pyxinol (821 mg, 1.72 mmol) was dissolved in anhydrous dichloromethane (20 mL), pyridinium chlorochromate (389 mg, 1.80 mmol) was added, and the mixture was stirred at room temperature for 12 h, filtered, concentrated in vacuo, and then subjected to silica gel column chromatography (SGCC) to obtain intermediate product 4. Compound 4 (26 mg, 0.054 mmol) was dissolved in 0.5 mL of dichloromethane, m-CPBA (101 mg, 0.438 mmol) was added, and the mixture was stirred at room temperature overnight, filtered, concentrated in vacuo, and then subjected to silica gel column chromatography (SGCC) to obtain intermediate product 6. Compound 6 (22 mg, 0.045 mmol) was dissolved in anhydrous dichloromethane (0.9 mL), P-TSA (68.3 mg, 0.359 mmol) was added, and the mixture was stirred at room temperature for 12 h, filtered, concentrated in vacuo, and then subjected to silica gel column chromatography (SGCC) to obtain intermediate product 8. Under argon protection, compound 8 (250 mg, 0.510 mmol) and 4-pyridinemethanol (0.612 mmol) were dissolved in anhydrous DCM (1 mL), EDCI (250 mg, 1.31 mmol) and DMAP (10 mg, 0.08 mmol) were added at 0 °C, and stirred at room temperature for about 30 minutes. Then, an appropriate amount of deionized water was added for quenching, and the organic layer was extracted with EA, and then saturated NaHCO3 The organic layer was washed with saturated NaCl solution and then with Na 2 SO 4 The residue was dried, concentrated and purified by column chromatography to obtain the target compound 12a [(20S,24S)-epoxy-3,4-oxygenase-3-(4-pyridinemethanol-1-yl)carbonyl-4(28)-ene-dammarane-12β,25-diol].

[0103] 1 H NMR (400 MHz, CDCl 3 )δ8.65(dd,J=4.4,1.6Hz,2H),7.31(dd,J=4.5,1.6Hz,2H),5.12(s,2H),4.86( t,J=1.8Hz,1H),4.67(d,J=1.3Hz,1H),3.88(dd,J=10.9,5.3Hz,1H),3.55(td,J =10.4,4.7Hz,1H),2.51–2.21(m,3H),2.14–0.83(m,19H),1.74(d,J=0.3Hz,3H) ,1.28(s,3H),1.23(s,3H),1.11(s,3H),1.06(s,3H),0.93(s,3H),0.91(s,3H).

[0104] Example 9: (20S, 24S)-epoxy-3,4-opening-3-(3-tetrahydrofuranol-1-yl)carbonyl-4(28)-ene-dammarane-12β,25-diol (12b);

[0105] Under argon protection, the intermediate product 8 in Example 14 (250 mg, 0.510 mmol) and 3-hydroxytetrahydrofuran (0.612 mmol) were dissolved in anhydrous DCM (10 mL), and EDCI (250 mg, 1.31 mmol) and DMAP (10 mg, 0.08 mmol) were added at 0 ° C. The mixture was stirred at room temperature for about 30 minutes, and then an appropriate amount of deionized water was added for quenching. The organic layer was extracted with EA, and then saturated NaHCO 3 The organic layer was washed with saturated NaCl solution and then with Na 2 SO 4 The residue was dried, concentrated and purified by column chromatography to obtain the target compound 12b [(20S,24S)-epoxy-3,4-opened-3-(3-tetrahydrofuranol-1-yl)carbonyl-4(28)-ene-dammarane-12β,25-diol].

[0106] 1 H NMR (400 MHz, CDCl 3)δ5.27(ddt,J=6.6,5.0,1.7Hz,1H),4.86(t,J=1.8Hz,1H),4.67(d,J=1.8Hz,1H),3.95–3.75(m,5H),3.55(td,J=10.4,4. 6Hz,1H),2.42–0.83(m,24H),1.74(s,3H),1.28(s,3H),1.24(s,3H),1.11(s,3H),1.06(s,3H),0.93(s,3H),0.89(s,3H).

[0107] Example 10: (20S, 24R)-epoxy-3,4-opened-3-(2-thiazolylmethanol-1-yl)carbonyl-dammarane-4,12β,25-triol (13a);

[0108] The intermediate product 5 (200 mg, 0.41 mmol) synthesized in Example 1 was dissolved in THF / H 2 O in a stirred solution (14 mL, 20:1, v / v), and KOH (110 mg, 2.03 mmol) was slowly added thereto. After stirring at room temperature for 12 h, it was filtered and concentrated in vacuo, and then silica gel column chromatography (SGCC) was used to obtain the intermediate compound intermediate product 9. Under argon protection, compound 9 (250 mg, 0.510 mmol) and 2-thiazolemethanol (0.612 mmol) were dissolved in anhydrous DCM (10 mL), and EDCI (250 mg, 1.31 mmol) and DMAP (10 mg, 0.08 mmol) were added at 0°C. The mixture was stirred at room temperature for about 30 minutes, and then an appropriate amount of deionized water was added for quenching. The organic layer was extracted with EA, and then saturated NaHCO 3 The organic layer was washed with saturated NaCl solution and then with Na 2 SO 4 The mixture was dried and concentrated, and the concentrated sample was purified by silica gel column chromatography. According to the adsorption properties of the target product on silica gel, a suitable eluent was selected for purification to obtain the target compound 13a [(20S, 24R)-epoxy-3,4-opened-3-(2-thiazolylmethanol-1-yl)carbonyl-dammarane-4,12β,25-triol].

[0109] 1 H NMR (400 MHz, CDCl 3)δ7.77(dd,J=10.6,3.1Hz,1H),7.34(dd,J=14.6,3.1Hz,1H),5.38(s,2H),3.85(dd ,J=8.8,6.5Hz,1H),3.50(td,J=10.5,4.3Hz,1H),2.57(tdd,J=15.2,10.6,4.4Hz,2H ),2.27(ddd,J=18.3,9.7,4.9Hz,1H),2.22–2.15(m,1H),2.12–0.84(m,18H),1.27(s ,6H),1.27(s,3H),1.24(s,3H),1.10(s,3H),1.02(s,3H),1.00(s,3H),0.88(s,3H).

[0110] Example 11: (20S, 24R)-epoxy-3,4-opened-3-(2-pyridinemethanol-1-yl)carbonyl-dammarane-4,12β,25-triol (13b);

[0111] Under argon protection, the intermediate product 9 in Example 10 (250 mg, 0.510 mmol) and 2-pyridinemethanol (0.612 mmol) were dissolved in anhydrous DCM (10 mL), and EDCI (250 mg, 1.31 mmol) and DMAP (10 mg, 0.08 mmol) were added at 0 ° C. The mixture was stirred at room temperature for about 30 minutes, and then an appropriate amount of deionized water was added for quenching. The organic layer was extracted with EA, and then saturated NaHCO 3 The organic layer was washed with saturated NaCl solution and then with Na 2 SO 4 The concentrated sample was purified by silica gel column chromatography. According to the adsorption properties of the target product on silica gel, a suitable eluent was selected for purification to obtain the target compound 13b [(20S, 24R)-epoxy-3,4-opened-3-(2-pyridinemethanol-1-yl)carbonyl-dammarane-4,12β,25-triol].

[0112] 1 H NMR (400 MHz, CDCl 3)δ8.60(d,J=4.5Hz,1H),7.73(td,J=8.0,1.9Hz,1H),7.37(d,J=7.8Hz,1H),7.24(dd,J=7.6,5 .0Hz,1H),5.23(s,2H),3.85(dd,J=8.7,6.5Hz,1H),3.51(td,J=10.5,4.5Hz,1H),2.71–2.51(m ,2H),2.30(ddd,J=15.4,11.3,6.4Hz,1H),2.19(ddd,J=10.9,9.0,3.8Hz,1H),2.11–0.83(m,18 H),1.28(s,3H),1.27(s,6H),1.24(s,3H),1.10(s,3H),1.03(s,3H),1.00(s,3H),0.88(s,3H).

[0113] Example 12: (20S, 24R)-epoxy-3,4-opened-3-(4-pyridinemethanol-1-yl)carbonyl-dammarane-4,12β,25-triol (13c);

[0114] Under argon protection, the intermediate product 9 in Example 10 (250 mg, 0.510 mmol) and 4-pyridinemethanol (0.612 mmol) were dissolved in anhydrous DCM (10 mL), and EDCI (250 mg, 1.31 mmol) and DMAP (10 mg, 0.08 mmol) were added at 0 ° C. The mixture was stirred at room temperature for about 30 minutes, and then an appropriate amount of deionized water was added for quenching. The organic layer was extracted with EA, and then saturated NaHCO 3 The organic layer was washed with saturated NaCl solution and then with Na 2 SO 4 The mixture was dried and concentrated, and the concentrated sample was purified by silica gel column chromatography. According to the adsorption properties of the target product on silica gel, a suitable eluent was selected for purification to obtain the target compound 13c [(20S,24R)-epoxy-3,4-opened-3-(4-pyridinemethanol-1-yl)carbonyl-dammarane-4,12β,25-triol].

[0115] 1 H NMR (400 MHz, CDCl 3)δ8.70(d,J=5.5Hz,2H),7.39(d,J=5.9Hz,2H),5.14(d,J=2.2Hz,2H),3.86 (dd,J=8.7,6.6Hz,1H),3.52(td,J=10.5,4.4Hz,1H),2.57(td,J=13.5,4.4H z,2H),2.35–2.15(m,2H),2.11–0.83(m,18H),1.28(s,3H),1.27(s,3H),1. 27(s,3H),1.25(s,3H),1.10(s,3H),1.03(s,3H),1.01(s,3H),0.90(s,3H).

[0116] Example 13: (20S, 24R)-epoxy-3,4-opened-3-(4-pyrazinemethanol-1-yl)carbonyl-dammarane-4,12β,25-triol (13d);

[0117] Under argon protection, the intermediate product 9 in Example 10 (250 mg, 0.510 mmol) and 4-pyrazinemethanol (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 at 0 ° C. The mixture was stirred at room temperature for about 30 minutes, and then an appropriate amount of deionized water was added for quenching. The organic layer was extracted with EA, and then saturated NaHCO 3 The organic layer was washed with saturated NaCl solution and then with Na 2 SO 4 The mixture was dried and concentrated, and the concentrated sample was purified by silica gel column chromatography. According to the adsorption properties of the target product on silica gel, a suitable eluent was selected for purification to obtain the target compound 13d [(20S, 24R)-epoxy-3,4-opened-3-(4-pyrazinemethanol-1-yl)carbonyl-dammarane-4,12β,25-triol].

[0118] 1 H NMR (400 MHz, CDCl 3)δ8.67(s,1H),8.56(dt,J=10.3,2.3Hz,2H),5.26(d,J=3.3Hz,2H),3.85(dd,J= 8.6,6.5Hz,1H),3.51(td,J=10.6,4.4Hz,1H),2.67–2.52(m,2H),2.30(ddd,J=15 .2,11.2,6.1Hz,1H),2.19(tt,J=9.2,4.0Hz,1H),2.12–0.83(m,18H),1.27(s,3H ),1.27(s,6H),1.25(s,3H),1.10(s,3H),1.03(s,3H),1.00(s,3H),0.89(s,3H).

[0119] Example 14: (20S, 24S)-epoxy-3,4-opened-3-(furfuryl-1-yl)carbonyl-dammarane-4-12β,25-triol (14a);

[0120] The intermediate product 6 (200 mg, 0.41 mmol) synthesized in Example 8 was dissolved in THF / H 2 O in a stirred solution (14 mL, 20:1, v / v), and KOH (110 mg, 2.04 mmol) was slowly added thereto. After stirring at room temperature for 12 h, it was filtered and concentrated in vacuo, and then silica gel column chromatography (SGCC) was used to obtain intermediate 10. Under argon protection, compound 10 (250 mg, 0.510 mmol) and furfuryl alcohol (0.612 mmol) were dissolved in anhydrous DCM (10 mL), and EDCI (250 mg, 1.31 mmol) and DMAP (10 mg, 0.08 mmol) were added at 0°C. The mixture was stirred at room temperature for about 30 minutes, and then an appropriate amount of deionized water was added for quenching. The organic layer was extracted with EA, and then saturated NaHCO 3 The organic layer was washed with saturated NaCl solution and then with Na 2 SO 4 The mixture was dried and concentrated, and the concentrated sample was purified by silica gel column chromatography. According to the adsorption properties of the target product on silica gel, a suitable eluent was selected for purification to obtain the target compound 14a [(20S, 24S)-epoxy-3,4-opened-3-(furfuryl-1-yl)carbonyl-dammarane-4,12β,25-triol].

[0121] 1 H NMR (400 MHz, CDCl 3)δ7.42(dd,J=1.9,0.8Hz,1H),6.40(dd,J=3.3,0.8Hz,1H),6.35(dd,J=3.3,1.8Hz,1H),5.05(d,J =7.9Hz,2H),3.88(dd,J=10.9,5.4Hz,1H),3.50(td,J=10.4,4.7Hz,1H),2.58(ddd,J=14.9,10.3, 4.8Hz,1H),2.47(ddd,J=15.3,10.2,4.8Hz,1H),2.24(tdd,J=10.3,6.9,5.1Hz,2H),2.12–0.85(m ,18H),1.27(s,6H),1.24(s,3H),1.23(s,3H),1.10(s,3H),1.05(s,3H),1.02(s,3H),0.87(s,3H).

[0122] Example 15: (20S, 24S)-epoxy-3,4-opened-3-(2-thiazolylmethanol-1-yl)carbonyl-dammarane-4,12β,25-triol (14b);

[0123] Under argon protection, the intermediate 10 (250 mg, 0.510 mmol) synthesized in Example 14 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 at 0 ° C. The mixture was stirred at room temperature for about 30 minutes, and then an appropriate amount of deionized water was added for quenching. The organic layer was extracted with EA, and then saturated NaHCO 3 The organic layer was washed with saturated NaCl solution and then with Na 2 SO 4 The mixture was dried and concentrated, and the concentrated sample was purified by silica gel column chromatography. According to the adsorption properties of the target product on silica gel, a suitable eluent was selected for purification to obtain the target compound 14b [(20S, 24S)-epoxy-3,4-opened-3-(2-thiazolylmethanol-1-yl)carbonyl-dammarane-4,12β,25-triol].

[0124] 1 H NMR (400 MHz, CDCl 3)δ7.79(d,J=3.3Hz,1H),7.36(d,J=3.2Hz,1H),5.39(s,2H),3.88(dd,J=10.8,5.4Hz,1H),3.53(td,J=10.4,4.6Hz,1H),2.58(tdd,J=15.3,10.6 ,4.5Hz,2H),2.37–2.22(m,2H),2.11–0.83(m,18H),1.28(s,6H),1.25( s,3H),1.23(s,3H),1.11(s,3H),1.06(s,3H),1.03(s,3H),0.90(s,3H).

[0125] Example 16: (20S, 24S)-epoxy-3,4-opened-3-(2-pyridinemethanol-1-yl)carbonyl-dammarane-4,12β,25-triol (14c);

[0126] Under argon protection, the intermediate product 10 (250 mg, 0.510 mmol) synthesized in Example 14 and 2-pyridinemethanol (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 at 0 ° C. The mixture was stirred at room temperature for about 30 minutes, and then an appropriate amount of deionized water was added for quenching. The organic layer was extracted with EA, and then saturated NaHCO 3 The organic layer was washed with saturated NaCl solution and then with Na 2 SO 4 The mixture was dried and concentrated, and the concentrated sample was purified by silica gel column chromatography. According to the adsorption properties of the target product on silica gel, a suitable eluent was selected for purification to obtain the target compound 14c [(20S, 24S)-epoxy-3,4-opened-3-(2-pyridinemethanol-1-yl)carbonyl-dammarane-4,12β,25-triol].

[0127] 1 H NMR (400 MHz, CDCl 3)δ8.60(d,J=4.9Hz,1H),7.76–7.67(m,1H),7.36(d,J=7.8Hz,1H),7.24(dd,J=7.3,4.9Hz,1 H),5.23(s,2H),3.88(dd,J=10.8,5.4Hz,1H),3.53(td,J=10.4,4.6Hz,1H),2.70–2.52(m,2H ),2.34(ddd,J=18.3,8.4,4.5Hz,1H),2.25(ddd,J=13.8,8.6,3.5Hz,1H),2.12–0.83(m,18H) ,1.28(s,6H),1.25(s,3H),1.23(s,3H),1.11(s,3H),1.06(s,3H),1.04(s,3H),0.90(s,3H).

[0128] Example 17: (20S, 24S)-epoxy-3,4-opened-3-(4-pyridinemethanol-1-yl)carbonyl-dammarane-4,12β,25-triol (14d);

[0129] Under argon protection, the intermediate 10 (250 mg, 0.510 mmol) synthesized in Example 14 and 4-pyridinemethanol (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 at 0 ° C. The mixture was stirred at room temperature for about 30 minutes, and then an appropriate amount of deionized water was added for quenching. The organic layer was extracted with EA, and then saturated NaHCO 3 The organic layer was washed with saturated NaCl solution and then with Na 2 SO 4 The reaction mixture was dried and concentrated, and the concentrated sample was purified by silica gel column chromatography. According to the adsorption properties of the target product on silica gel, a suitable eluent was selected for purification to obtain the target compound 14d [(20S, 24S)-epoxy-3,4-opened-3-(4-pyridinemethanol-1-yl)carbonyl-dammarane-4,12β,25-triol].

[0130] 1 H NMR (400 MHz, CDCl 3)δ8.62(d,J=5.1Hz,2H),7.29(d,J=5.4Hz,2H),5.12(s,2H),3.88(dd,J=10.9,5.3Hz,1H),3.54(td,J=10.4,4.6Hz,1H),2.64–2.52(m,2H),2. 37–2.22(m,2H),2.12–0.83(m,18H),1.28(s,3H),1.27(s,3H),1.25(s ,3H),1.24(s,3H),1.11(s,3H),1.06(s,3H),1.04(s,3H),0.91(s,3H).

[0131] Example 18: (20S, 24S)-epoxy-3,4-opened-3-(2-pyrazinemethanol-1-yl)carbonyl-dammarane-4,12β,25-triol (14e);

[0132] Under argon protection, the intermediate product 10 (250 mg, 0.510 mmol) synthesized in Example 14 and 2-pyrazinemethanol (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 at 0 ° C. The mixture was stirred at room temperature for about 30 minutes, and then an appropriate amount of deionized water was added for quenching. The organic layer was extracted with EA, and then saturated NaHCO 3 The organic layer was washed with saturated NaCl solution and then with Na 2 SO 4 The mixture was dried and concentrated, and the concentrated sample was purified by silica gel column chromatography. According to the adsorption properties of the target product on silica gel, a suitable eluent was selected for purification to obtain the target compound 14e [(20S, 24S)-epoxy-3,4-opened-3-(2-pyrazinemethanol-1-yl)carbonyl-dammarane-4,12β,25-triol].

[0133] 1 H NMR (400 MHz, CDCl 3)δ8.67(d,J=1.5Hz,1H),8.56(dd,J=2.5,1.5Hz,1H),8.54(d,J=2.6Hz,1H), 5.26(d,J=3.1Hz,2H),3.88(dd,J=10.9,5.4Hz,1H),3.53(td,J=10.4,4.6Hz, 1H),2.68–2.53(m,2H),2.39–2.20(m,2H),2.14–0.83(m,18H),1.28(s,6H),1 .25(s,3H),1.23(s,3H),1.10(s,3H),1.06(s,3H),1.04(s,3H),0.90(s,3H).

[0134] Example 19: Cytotoxicity Evaluation of PPD Derivatives

[0135] Experimental methods:

[0136] (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 incubated at 37°C, 5% CO 2 Culture in a cell culture incubator;

[0137] (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;

[0138] (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%

[0139] Experimental results: When the drug concentration was 30 μM, the survival rate of tumor-resistant KBV cells treated with Vrp was about 40%, while the survival rates of tumor-resistant KBV cells treated with compounds 11a-11g, 12a-12b, 13a-13d, and 14a-14e were all above 80%, indicating that the cytotoxicity of these compounds was significantly lower than that of Vrp.

[0140] Table 1 Cytotoxicity evaluation results of PPD derivatives in KBV cells

[0141]

[0142]

[0143] Example 20: Evaluation of the anti-tumor MDR activity of PPD derivatives

[0144] Experimental methods:

[0145] (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 incubated at 37°C, 5% CO 2 Culture in a cell culture incubator;

[0146] (2) Cell administration: After the cells were plated and adhered for 24 hours, 10 μM of different derivatives, 100 nM of paclitaxel and DMSO were added as controls, 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;

[0147] (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%

[0148] Experimental results: Paclitaxel has no cytotoxicity to the tumor-resistant cell line KBV at 100nM. When sapogenin PPD and parent compound 7-10 were added at a concentration of 10μM, cytotoxicity was still not shown, indicating that at this concentration, these compounds failed to increase the sensitivity of paclitaxel-resistant tumor-resistant cells KBV to paclitaxel. When PPD derivatives 11a-11g, 12a-12b, 13a-13d, and 14a-14e were added in combination, the survival rate of KBV cells was significantly reduced, indicating that they significantly increased the sensitivity of paclitaxel-resistant tumor-resistant cells KBV to paclitaxel, that is, they showed a tumor resistance reversal effect. Among them, the tumor resistance reversal activity of the preferred compounds 11a, 11b, 11c, 11d, 11f, 11g, 12a, 12b, 13a, 13b, 13c, 14a and 14d was significantly better than that of the positive control drug verapamil (Vrp).

[0149] Table 2 Evaluation results of the anti-tumor MDR activity of PPD derivatives on KBV cells

[0150]

[0151]

[0152] The above description is only a partial embodiment of the present application and an illustration of the technical principles used.

[0153] 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 derivative, whose structural formula is shown in Formula 11, 12, 13 or 14, or a pharmaceutically acceptable salt of the compound shown in Formula 11, 12, 13 or 14, in, R1, R2, R3, and R4 are each independently R x -KY-, where Y represents NH or O, K represents methylene or is absent, 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 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 derivative according to claim 1, characterized in that R1 represents 2-pyrazinemethylamine-1-yl, 4-pyridinemethanol-1-yl, 3-N-Boc-azetidinol-1-yl, 3-azetidinol-1-yl, 2-thiophenemethylamine-1-yl, 3-N-Boc-azetidinamine-1-yl or 4-N-Boc-piperidinol-1-yl; R2 represents 4-pyridinemethanol-1-yl or 3-tetrahydrofuranol-1-yl; R3 represents 2-thiazolylmethanol-1-yl, 2-pyridinemethanol-1-yl, 4-pyridinemethanol-1-yl or 2-pyrazinemethanol-1-yl; R4 represents furfuryl-1-yl, 2-thiazolylmethanol-1-yl, 2-pyridinemethanol-1-yl, 4-pyridinemethanol-1-yl or 2-pyrazinemethanol-1-yl.

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

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

6. Use of the PPD ring-opened 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 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 open-ring 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 open-ring derivatives and their pharmaceutically acceptable salts shown in Formula 11, 12, 13 or 14.

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 PPD ring-opening derivative according to any one of claims 1 to 4 and a pharmaceutically acceptable salt thereof.

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