ANILINO PODOPHYLLIN DERIVATIVE HAVING ANTITUMOR ACTIVITY, METHOD FOR PREPARATION THEREOF, AND USE THEREOF

• US 20160289242A1
• Filed: 09/05/2014
• Published: 10/06/2016
• Est. Priority Date: 11/15/2013
• Status: Active Grant

• Current Assignee: Hebei University of Technology
• Original Assignee: Hebei University of Technology
• Inventors: YANG, Yong, TANG, Yajie, ZHAO, Wei, LI, Hongmei
• Granted Patent: US 9,714,254 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A61K 31/365 Lactones
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07D 493/04 Ortho-condensed systems
• CPC Class Codes: Y02P 20/582 Recycling of unreacted starting or intermediate materials

Abstract

The present invention discloses an anilino-substituted podophyllin derivative having antitumor activity, method for preparation thereof, and use thereof. By means of anilino reactions, the present invention introduces 4-chloro-3-methylaniline, 3-fluoro-4-methoxyaniline, 4,4′-diaminodiphenylmethane, o-anisidine, 4-chloro-2-aminoanisole, anthranilonitrile, 2,6-dichloro-4-aminophenol, N,N-dimethylamino meta-aniline, 2-ethyl-5-nitroaniline, 2 2′-diaminodiphenylsulfide, or 2-aminobenzotrifluoride into position 4 of the active C-ring of podophyllotoxin or 4′-demethylepipodophyllotoxin to obtain the aniline-substituted podophyllotoxin derivative shown in formula (V); by means of multi-pathway and multi-target effects on tumor cells, said derivative has significantly increased antitumor activity, and can be prepared as an antineoplastic drug and applied in clinical antitumor therapy.

Claims

1-10. -10. (canceled)

11. Aniline-substituted podophyllotoxin-type derivatives with anti-tumor activity or sails thereof, with structural formula (V):

12. A method for preparing the aniline-substituted podophyllotoxin-type derivatives as claimed in claim 11, comprising the steps of:

by aniline substitution reaction, 4-chloro-3-methylaniline, 3-fluoro-4-methoxyaniline, 4,4′

-diaminodiphenylmethane, o-anisidine, 4-chloro-2-aminoanisole, o-aminobenzonitrile, 2,6-dichloro-4-aminophenol, N,N-dimethylamino metanil, 2-ethyl-5-nitroaniline, 2 2′

-diaminodiphenylsulfide or 2-aminobenzotrifluoride is introduced into position 4 of C-ring of podophyllotoxin and 4′

-demethylepipodophyllotoxin respectively, to get crude product of aniline-substituted podophyllotoxin-type derivatives.

13. The method as claimed in claim 12, wherein, the aniline reaction is comprising:

(1) activating position 4 of C-ring of podophyllotoxin or 4′

-demethylepipodophyllotoxin;

(2) podophyllotoxin or 4′

-demethylepipodophyllotoxin with activated position 4 of C-ring being dissolved in organic solvent, then adding 4-chloro-3-methylaniline, 3-fluoro-4-methoxyaniline, 4,4′

-diaminodiphenylmethane, o-aminoanisole, 4-chloro-2-aminoanisole, o-aminobenzonitrile, 2,6-dichloro-4-aminophenol, N,N-dimethylamino metanil, 2-ethyl-5-nitroaniline, 2 2′

-diaminodiphenylsulfide or 2-aminobenzotrifluoride respectively;

stirring to carried out the aniline reaction.

14. The method as claimed in claim 13, wherein, manner of activation of position 4 of C-ring of podophyllotoxin and 4′

-demethylepipodophyllotoxin is to use hydrobromic acid to activate position 4 of C-ring of podophyllotoxin and 4′

-demethylepipodophyllotoxin;

preferably, the manner of activation includes steps of;

podophyllotoxin and 4′

-demethylepipodophyllotoxin being dried, and under protection of nitrogen, hydrobromic acid being added while stirring under ice-bath;

after the addition, removing ice-bath, then reacting under 20-25°

C. for 5-12 hours.

15. The method as claimed in claim 13, wherein, the organic solvent in step (2) is methylene chloride;

the stirring is carried out in vacuo with rotational speed of 50 to 800 rpm, preferred 600 rpm;

temperature of the aniline substitution reaction is 10-40°

C., reaction time of the aniline substitution reaction is 12-48 hours.

16. The method as claimed in claim 12, wherein, in the aniline substitution reaction, molar ratio between podophyllotoxin or 4′

-demethylepipodophyllotoxin and 4-chloro-3-methylaniline, 3-fluoro-4-methoxyaniline, 4,4′

-diaminodiphenylmethane, o-aminoanisole, 4-chloro-2-aminoanisole, o-aminobenzonitrile, 2,6-dichloro-4-aminophenol, N,N-dimethylamino metanil, 2-ethyl-5-nitroaniline, 2 2′

-diaminodiphenylsulfide or 2-aminobenzotrifluoride is preferably 1;

2.

17. The method as claimed in claim 12, further comprising:

(1) the crude product of aniline-substituted podophyllotoxin-type being subjected rotary evaporation and concentration, then extracted by methylene chloride/water with volume ratio of 1;

1 three times, to get sample to be separated and purificated;

(2) the sample to be separated and purificated being separated by silica gel column chromatography, gel column chromatography and high performance preparative liquid chromatography separations sequentially, to obtain clean product of compound of formula (V);

18. The method as claimed in claim 17, wherein separation method by silica gel column chromatography comprises:

(1) the silica gel column chromatography being normal phase silica gel column chromatography, wherein normal phase silica gel is mixed in organic solvent with low polarity, loaded into column, balanced with eluent which is formed from chloroform and acetone with volume ratio of 10;

1;

(2) samples being dissolved with the eluent, subjected to sample adsorption, then eluted with eluent which is collected later, then the sample being evaporated to dryness and recrystallized,

separation method by gel column chromatography comprises;

(1) soaking the gel with eluent which is formed from petroleum ether, chloroform and methanol with volume ratio of 5;

5;

1 in order;

loading processed gel into column and balanced with the eluent;

(2) sample preliminary separated by silica gel column chromatography being dissolved in the eluent, subjected to sample absorption, and then eluated with eluent which is collected later, then the sample being evaporated to dryness and recrystallized.

19. An anti-tumor pharmaceutical composition, comprising aniline-substituted podophyllotoxin-type derivatives as claimed in claim 11 with effective amount in treatment, or salts thereof, and a pharmaceutically acceptable carrier thereof.