Process of preparing chiral diacylhydrazine ligands
View Patent ↗The present invention provides diacylhydrazine ligands and chiral diacylhydrazine ligands for use with ecdysone receptor-based inducible gene expression systems. Thus, the present invention is useful for applications such as gene therapy, large scale production of proteins and antibodies, cell-based screening assays, functional genomics, proteomics, metabolomics, and regulation of traits in transgenic organisms, where control of gene expression levels is desirable. An advantage of the present invention is that it provides a means to regulate gene expression and to tailor expression levels to suit the user's requirements.
1. A process for preparing a compound of Formula IV:
wherein:
A is optionally substituted aryl or optionally substituted heteroaryl;
B is 3,5-di-methylphenyl;
R 2 is tert-butyl; and
R 8 , R 9 , R 10 , R 11 and R 12 are hydrogen;
wherein the asymmetric carbon atom to which R 2 is attached is enantiomerically enriched in the R isomer
comprising:
a) reacting a compound of Formula V:
with a compound of Formula VI:
R 2 HC═N—NH—CO 2 R 7 Formula VI
wherein:
X and Y are independently O or NR wherein R is alkyl or aryl;
C a and C b are independently an asymmetric carbon atom of the S configuration;
R 14 and R 15 are independently alkyl or aryl;
R 13 is halo, hydrogen, alkyl, alkoxy or OSO 2 CF 3 ;
R 7 is alkyl, arylalkyl or aryl;
R 2 is tert-butyl and
R 8 , R 9 , R 10 , R 11 and R 12 are hydrogen;
to form a compound of Formula VII:
b) reducing the compound of Formula VII to form a compound of Formula VIII:
c) reacting the compound of Formula VIII with a compound of Formula B-CO-LG wherein B is 3,5-di-methylphenyl and LG is a leaving group to form a compound of Formula IX:
d) removing the R 7 CO 2 — group of the compound of Formula IX to form a compound of Formula X; and
e) reacting the compound of Formula X with a compound of Formula A-CO-LG wherein A is optionally substituted aryl or optionally substituted heteroaryl and LG is a leaving group, to form a compound of Formula IV selected from the group consisting of:
(R)-3,5-dimethyl-benzoic acid N-(1-tert-butyl-butyl)-N′-(2-ethyl-3-methoxy-benzoyl)-hydrazide;
(R)-3,5-dimethyl-benzoic acid N′-benzoyl-N-(1-tert-butyl-butyl)-hydrazide;
(R)-3,5-dimethyl-benzoic acid N-(1-tert-butyl-butyl)-N′-(2-methyl-benzoyl)-hydrazide;
(R)-3,5-dimethyl-benzoic acid N-(1-tert-butyl-butyl)-N′-(2-fluoro-benzoyl)-hydrazide;
(R)-3,5-dimethyl-benzoic acid N-(2-bromo-benzoyl)-N-(1-tert-butyl-butyl)-hydrazide;
(R)-3,5-dimethyl-benzoic acid N-(1-tert-butyl-butyl)-N′-(3-methyl-benzoyl)-hydrazide;
(R)-3,5-dimethyl-benzoic acid N-(1-tert-butyl-butyl)-N′-(4-methyl-benzoyl)-hydrazide;
(R)-3,5-dimethyl-benzoic acid N-(1-tert-butyl-butyl)-N′-(2-chloro-pyridine-3-carbonyl)-hydrazide;
(R)-3,5-dimethyl-benzoic acid N-(1-tert-butyl-butyl)-N′-(6-chloro-pyridine-3-carbonyl)-hydrazide; and
(R)-3,5-dimethyl-benzoic acid N-(1-tert-butyl-butyl)-N′-(3-methoxy-2-methyl-benzoyl)-hydrazide.
2. The process of claim 1 , wherein said compound of Formula IV is (R)-3,5-dimethyl-benzoic acid N-(1-tert-butyl-butyl)-N′-(2-ethyl-3-methoxy-benzoyl)-hydrazide.
3. The process of claim 1 , wherein said compound of Formula IV is (R)-3,5-dimethyl-benzoic acid N′-benzoyl-N-(1-tert-butyl-butyl)-hydrazide.
4. The process of claim 1 , wherein said compound of Formula IV is (R)-3,5-dimethyl-benzoic acid N-(1-tert-butyl-butyl)-N′-(2-methyl-benzoyl)-hydrazide.
5. The process of claim 1 , wherein said compound of Formula IV is (R)-3,5-dimethyl-benzoic acid N-(1-tert-butyl-butyl)-N′-(2-fluoro-benzoyl)-hydrazide.
6. The process of claim 1 , wherein said compound of Formula IV is (R)-3,5-dimethyl-benzoic acid N′-(2-bromo-benzoyl)-N-(1-tert-butyl-butyl)-hydrazide.
7. The process of claim 1 , wherein said compound of Formula IV is (R)-3,5-dimethyl-benzoic acid N-(1-tert-butyl-butyl)-N′-(3-methyl-benzoyl)-hydrazide.
8. The process of claim 1 , wherein said compound of Formula IV is (R)-3,5-dimethyl-benzoic acid N-(1-tert-butyl-butyl)-N′-(4-methyl-benzoyl)-hydrazide.
9. The process of claim 1 , wherein said compound of Formula IV is (R)-3,5-dimethyl-benzoic acid N-(1-tert-butyl-butyl)-N′-(2-chloro-pyridine-3-carbonyl)-hydrazide.
10. The process of claim 1 , wherein said compound of Formula IV is (R)-3,5-dimethyl-benzoic acid N-(1-tert-butyl-butyl)-N′-(6-chloro-pyridine-3-carbonyl)-hydrazide.
11. The process of claim 1 , wherein said compound of Formula IV is (R)-3,5-dimethyl-benzoic acid N-(1-tert-butyl-butyl)-N′-(3-methoxy-2-methyl-benzoyl)-hydrazide.