Amooranin compounds and analogs thereof and related methods of use

1. A method for synthesizing 25-Hydroxy-3-oxoolean-12-en-28-oic Acid with chemical Structure

comprising the steps of:

(a) conducting a Jones oxidation of oleanolic acid with chemical structure

by adding a Jones reagent solution to the oleanolic acid suspended in CH 3 COCH 3 , and CHCl 3 or CH 2 Cl 2 , and then adding i-PrOH, H 2 O, and CH 2 Cl 2 , and then concentrating Jones oxidation organic layer to obtain 3-Oxoolean-12-en-28-oic Acid with chemical structure

(b) adding benzyl chloride and K 2 CO 3 to a solution of the 3-Oxoolean-12-en-28-oic acid in dioxane, and then removing solids from dioxane layer, and then concentrating the dioxane layer to obtain 3-Oxoolean-12-en-28-oic Acid Benzyl Ester with chemical structure

(c) refluxing a suspension of the 3-Oxoolean-12-en-28-oic Acid Benzyl Ester, toluene, ethylene glycol, and p-toluenesulfonic acid with azeotropic removal of water, and then washing resulting mixture with aqueous NaHCO 3 , and then concentrating resulting organic layer to obtain 3-(10,30-Dioxolane)olean-12-en-28-oic Acid Benzyl Ester with chemical structure

(d) oxidizing the 3-(10,30-Dioxolane)olean-12-en-28-oic Acid Benzyl Ester by adding NaIO 4 , Bu 4 NBr, and RuCl 3 to the 3-(10,30-Dioxolane)olean-12-en-28-oic Acid Benzyl Ester dissolved in CCl 4 and H 2 O, and then concentrating filtered organic layer to obtain 3-(10,30-Dioxolane)olean-11-oxo-12-en-28-oic Acid Benzyl Ester with chemical structure

(e) adding sodium triethylborohydride in THF to the 3-(10,30-Dioxolane)olean-11-oxo-12-en-28-oic Acid Benzyl Ester dissolved in dry THF, and then adding H 2 O, and then extracting with EtOAc, and then concentrating EtOAc extract to obtain 3-(10,30-Dioxolane)olean-11β-hydroxy-12-en-28-oic Acid Benzyl Ester with chemical structure

(f) adding nitrosyl chloride to the 3-(10,30-Dioxolane)olean-11β-hydroxy-12-en-28-oic Acid Benzyl Ester in pyridine, and then pouring mixture into ice-water to form a suspension, and then extracting the suspension with EtOAc, and then concentrating the EtOAc extract to obtain a nitrite compound with chemical structure

(g) irradiating the nitrite compound in dry toluene under a nitrogen atmosphere with a high pressure mercury lamp to obtain an aldoxime compound with chemical structure in toluene;

(h) removing the toluene in vacuo from the aldoxime compound, and then treating the aldoxime compound in a mixture of dioxane, water, and acetic acid, to sodium nitrite in water to form an aldehyde compound and a hemiacetal compound in the mixture, and then extracting the mixture with EtOAc and washing the EtOAC extract with saturated aqueous NaHCO 3 , and then concentrating the EtOAc extract to obtain the aldehyde compound and the hemiacetal compound;

(i) adding NaBH 4 in MeOH to the aldehyde compound and hemiacetal compound dissolved in CHCl 3 , and then concentrating organic layer to obtain 3-(10,30-Dioxolane)olean-11β,25-dihydroxy-12-en-28-oic Acid Benzyl Ester with chemical structure

(j) treating the 3-(10,30-Dioxolane)olean-11β,25-dihydroxy-12-en-28-oic Acid Benzyl Ester in a CHCl 3 and pyridine solution with acetic anhydride, and then washing the pyridine and CHCl 3 layer with aqueous NaHCO 3 and then concentrating the organic layer to obtain 3-(10,30-Dioxolane)olean-11β-hydroxy-25-acetoxy-12-en-28-oic Acid Benzyl Ester chemical structure

(k) hydrogenating the 3-(10,30-Dioxolane)olean-11β-hydroxy-25-acetoxy-12-en-28-oic Acid Benzyl Ester dissolved in NaHCO 3 -treated EtOH with 10% palladium on charcoal catalyst and then concentrating filtered reaction mixture to obtain a residue;

(l) treating the residue with 30% TFA in CH 2 Cl 2 , and then adding saturated aqueous NaHCO 3 , and then extracting with EtOAc and concentrating the EtOAc extract to obtain 3-Oxo-25-acetoxy-olean-12-en-28-oic Acid with chemical structure

and

(m) heating under reflux a mixture of the 3-Oxo-25-acetoxy-olean-12-en-28-olic Acid in methanol, with KOH in water or with NaOH in water, and then acidifying the mixture to form the 25-Hydroxy-3-oxoolean-12-en-28-oic Acid, and then extracting the mixture with EtOAc and then concentrating the EtOAc extract to obtain the 25-Hydroxy-3-oxoolean-12-en-28-oic Acid.

2. The method according to claim 1 , further comprising the step of:

(n) treating a solution of the 25-Hydroxy-3-oxoolean-12-en-28-oic Acid in ethyl ether with diazomethane, and then destroying excess diazomethane with acetic acid, and then washing resulting ethyl ether solution with a NaHCO 3 solution, and then concentrating the ethyl ether solution to obtain 25-Hydroxy-3-oxoolean-12-en-28-oic Acid Methyl Ester with chemical structure

3. The method according to claim 1 , wherein at step (a) the Jones reagent solution is added to the Oleanolic acid in the CH 3 COCH 3 , and the CH 2 Cl 2 .

4. The method according to claim 1 , wherein at step (a) the Jones reagent solution is added to the Oleanolic acid in the CH 3 COCH 3 , and the CHCl 3 .

5. The method according to claim 1 , wherein at step (a) wherein the Jones oxidation of the oleanolic acid is conducted at a temperature at 5° C.

6. The method according to claim 1 , further comprising at step (a) purifying the 3-Oxoolean-12-en-28-oic Acid by a crystallization of the 3-Oxoolean-12-en-28-oic Acid in methanol.

7. The method according to claim 1 , further comprising at step (b) refluxing mixture of the benzyl chloride, the K 2 CO 3 and the 3-Oxoolean-12-en-28-oic acid in the dioxane before removing the solids from the dioxane layer.

8. The method according to claim 1 , further comprising at step (b) crystallizing the 3-Oxoolean-12-en-28-oic Acid Benzyl Ester in CHCl 3 and MeOH.

9. The method according to claim 1 , wherein at step (c) a Dean-Stark apparatus is used for for the azeotropic removal of water during the refluxing of the suspension of the 3-Oxoolean-12-en-28-oic Acid Benzyl Ester, toluene, ethylene glycol, and p-toluenesulfonic acid.

10. The method according to claim 1 , further comprising at step (c) crystallizing the 3-(10,30-Dioxolane)olean-12-en-28-oic Acid Benzyl Ester in CHCl 3 and MeOH.

11. The method according to claim 1 ,

wherein at step (d) 0.06 molar:0.3 molar is the molar ratio of the 3-(10,30-Dioxolane)olean-12-en-28-oic Acid Benzyl Ester to the NaIO 4 , and

wherein at step (d) 0.06 molar:0.12 molar is the molar ratio of the 3-(10,30-Dioxolane)olean-12-en-28-oic Acid Benzyl Ester to the RuCl 3 .

12. The method according to claim 1 , further comprising at step (d) using saturated aqueous NaCl to wash the filtered organic layer before concentrating the filtered organic layer to obtain the 3-(10,30-Dioxolane)olean-11-oxo-12-en-28-oic Acid Benzyl Ester.

13. The method according to claim 1 , further comprising at step (d) crystallizing the 3-(10,30-Dioxolane)olean-11-oxo-12-en-28-oic Acid Benzyl Ester in CHCl 3 and MeOH.

14. The method according to claim 1 , wherein at step (e) the 3-(10,30-Dioxolane)olean-12-en-28-oic Acid Benzyl Ester is dissolved in dry −15° C. THF.

15. The method according to claim 1 , further comprising at step (e) using aqueous NaCl to wash the EtOAc extract before concentrating the EtOAc extract to obtain the 3-(10,30-Dioxolane)olean-11β-hydroxy-12-en-28-oic Acid Benzyl Ester.

16. The method according to claim 1 further comprising at step (e) using a silica gel chromatography with hexane and acetone to purify the 3-(10,30-Dioxolane)olean-11β-hydroxy-12-en-28-oic Acid Benzyl Ester.

17. The method according to claim 1 wherein at step (f) the nitrosyl chloride is generated from NaNO 2 and HCl.

18. The method according to claim 1 wherein at step (f) the nitrosyl chloride is added to a solution of the 3-(10,30-Dioxolane)olean-11β-hydroxy-12-en-28-oic Acid Benzyl Ester in pyridine at −35° C.

19. The method according to claim 1 wherein at step (g) the irradiating of the nitrite compound occurs at a room temperature.

20. The method according to claim 1 wherein at step (h) the treating of the aldoxime compound in the mixture of dioxane, water, and acetic acid, to the sodium nitrite in water occurs at 0° C.

21. The method according to claim 1 further comprising at step (i) using a silica gel chromatography with hexane and acetone to purify the 3-(10,30-Dioxolane)olean-11β,25-dihydroxy-12-en-28-oic Acid Benzyl Ester.

22. The method according to claim 1 wherein step (j) at the treating of the 3-(10,30-Dioxolane)olean-11β,25-dihydroxy-12-en-28-oic Acid Benzyl Ester in the CHCl 3 and pyridine solution with the acetic anhydride occurs at a room temperature.

23. The method according to claim 1 further comprising at step (j) using a silica gel chromatography with hexane and acetone to purify the 3-(10,30-Dioxolane)olean-11β-hydroxy-25-acetoxy-12-en-28-oic Acid Benzyl Ester.

24. The method according to claim 1 wherein at step (k) the hydrogenating of the 3-(10,30-Dioxolane)olean-11β-hydroxy-25-acetoxy-12-en-28-oic Acid Benzyl Ester dissolved in NaHCO 3 -treated EtOH with 10% palladium on charcoal catalyst occurs at a room temperature.

25. The method according to claim 1 further comprising at step (l) purifying the 3-Oxo-25-acetoxy-olean-12-en-28-oic Acid by a silica gel chromatography using petroleum ether and acetone.

26. The method according to claim 1 wherein at step (m) aqueous HCl is used for acidifying the mixture after the heating under reflux.

27. The method according to claim 1 further comprising at step (m) washing the EtOAc extracts with saturated aqueous NaCl and drying the washed EtOAc extracts over MgSO 4 before concentrating the EtOAc extracts to obtain the 25-Hydroxy-3-oxoolean-12-en-28-oic Acid.

28. The method according to claim 1 , further comprising at step (m) purifying the 25-Hydroxy-3-oxoolean-12-en-28-oic Acid by a silica gel chromatography using hexane and acetone.

29. The method according to claim 2 wherein the esterifying of the 25-Hydroxy-3-oxoolean-12-en-28-oic Acid occurs in the ethyl ether cooled to 0° C.

30. The method according to claim 2 further comprising purifying the 25-Hydroxy-3-oxoolean-12-en-28-oic Acid Methyl Ester by a silica gel chromatography using petroleum ether and acetone.