METHODS AND INTERMEDIATES FOR THE PREPARATION OF BILE ACID DERIVATIVES

1 . A method of preparing a compound of formula I

or pharmaceutically acceptable salt, hydrate, solvate or amino acid, sulfate or glucuronide conjugate, or prodrug thereof,

wherein:

R 1 is OH, alkoxy, or oxo;

R 2 and R 3 are each independently H, OH, OSO 3 H, OCOCH 3 , OPO 3 H 2 , halogen, or alkyl optionally substituted with one or more halogen or OH, or R 2 and R 3 taken together with the carbon atom to which they are attached form a carbonyl;

R 4 is H, halogen, alkyl optionally substituted with one or more halogen or OH, alkenyl, or alkynyl;

R 5 and R 6 are each independently H, OH, OSO 3 H, OCOCH 3 , OPO 3 H 2 , halogen, or alkyl optionally substituted with one or more halogen or OH, or R 5 and R 6 taken together with the carbon atom to which they are attached form a carbonyl;

R 7 is OH, OSO 3 H, SO 3 H, OSO 2 NH 2 , SO 2 NH 2 , OPO 3 H 2 , PO 3 H 2 , CO 2 H, C(O)NHOH, NH(CH 2 ) 2 SO 3 H, NHCH 2 CO 2 H or optionally substituted tetrazolyl, oxadiazolyl, thiadiazolyl, 5-oxo-1,2,4-oxadiazolyl, 5-oxo-1,2,4-thiadiazolyl, oxazolidinedionyl, thiazolidine-dionyl, 3-hydroxyisoxazolyl, 3-hydroxyisothiazolyl, pyrimidine, 3,5-difluoro-4-hydroxyphenyl or 2,4-difluoro-3-hydroxyphenyl;

R 8 , R 9 , and R 10 are each independently H, OH, halogen, or alkyl optionally substituted with one or more halogen or OH, or R 8 and R 9 taken together with the carbon atoms to which they are attached form a 3- to 6-membered carbocyclic or heterocyclic ring comprising 1 or 2 heteroatoms selected from N, O, and S, or R 9 and R 10 taken together with the carbon atoms to which they are attached form a 3- to 6-membered carbocyclic or heterocyclic ring comprising 1 or 2 heteroatoms selected from N, O, and S;

m is 0, 1, or 2;

n is 0 or 1; and

p is 0 or 1;

the method comprising the step of reacting a compound of formula I-4 with a halogenating reagent to provide a compound of formula I-5a

wherein X is —(CHR 8 ) m (CHR 9 ) n (CHR 10 ) p —R 7 , where R 7 , R 8 , R 9 , and R 10 may be protected by R 11 or another protecting group.

2 . The method of claim 1 wherein compound of formula I is compound of formula I-9

or pharmaceutically acceptable salt, hydrate, solvate or amino acid, sulfate or glucuronide conjugate, or prodrug thereof.

3 . The method of claim 1 wherein compound of formula I is compound of formula II

or pharmaceutically acceptable salt, hydrate, solvate or amino acid, sulfate or glucuronide conjugate, or prodrug thereof.

4 . The method of claim 1 wherein compound of formula I is compound of formula III

or pharmaceutically acceptable salt, hydrate, solvate or amino acid, sulfate or glucuronide conjugate, or prodrug thereof, where is R 2 and R 5 and OH.

5 . The method of claim 1 wherein compound of formula I is compound of formula 100

6 . The method of claim 1 , wherein halogenating agent is a brominating agent.

7 . The method of claim 1 , wherein halogenating reagent is an iodinating reagent.

8 . The method of claim 1 further comprising converting compound of formula I-1 into compound of formula I-4 comprising the steps of

1) protecting the compound of formula I-1 to provide compound of formula I-2;

2) forming a leaving group at C12 to provide compound of formula I-3; and

3) eliminating the leaving group at C12 to provide the alkene compound of formula I-4.

9 . The method of claim 1 further comprising converting compound of formula I-5a into compound of formula I-6a comprising the steps of

5) reacting the compound of formula I-5a with an oxidizing agent to prepare a compound of formula I-6a.

10 . The method of claim 9 further comprising converting compound of formula I-6a into compound of formula I-7 comprising the steps of

6) reacting the compound of formula I-6a with a reducing agent to prepare the compound of formula I-7.

11 . The method of claim 10 further comprising converting compound of formula I-7 into compound of formula I comprising the steps of

7) optionally deprotecting the compound of formula I-7 to obtain the compound of formula I-8 and

8) reacting the compound of formula I-7 or I-8 with a reducing agent to provide the compound of formula I.

12 . The method of claim 1 further comprising converting compound of formula I-5 or I-5b into compound of formula I comprising the steps of:

4) reacting compound of formula I-5b with a reducing agent to prepare the compound of formula I-5c;

5) reacting the compound of formula I-5c with a reducing agent to provide the compound of formula I-5d; and

6) deprotecting the compound of formula I-5d to provide the compound of formula I.

13 . The method of claim 10 further comprising converting compound of formula I-7 or I-7a into compound of formula I comprising the steps of:

3) reacting the compound of formula I-7a with a reducing agent to provide the compound of formula I-8a;

4) deprotecting the compound of formula I-8a to obtain the compound of formula I.

14 . The method of claim 6 wherein the brominating agent is N-bromosuccinimide.

15 . The method of claim 7 wherein the iodinating agent is N-iodosuccinimide.

16 . The method of claim 9 wherein the oxidizing agent is RuCl 3 .

17 . The method of claim 10 wherein the reducing agent is zinc metal.

18 . The method of claim 11 wherein the reducing agent is sodium borohydride.

19 . The method of preparing a compound of formula D5

or pharmaceutically acceptable salt, hydrate, solvate or amino acid, sulfate or glucuronide conjugate, or prodrug thereof, wherein:

R 7 is OH, OSO 3 H, SO 3 H, OSO 2 NH 2 , SO 2 NH 2 , OPO 3 H 2 , PO 3 H 2 , CO 2 H, C(O)NHOH, NH(CH 2 ) 2 SO 3 H, NHCH 2 CO 2 H or optionally substituted tetrazolyl, oxadiazolyl, thiadiazolyl, 5-oxo-1,2,4-oxadiazolyl, 5-oxo-1,2,4-thiadiazolyl, oxazolidine-dionyl, thiazolidine-dionyl, 3-hydroxyisoxazolyl, 3-hydroxyisothiazolyl, pyrimidine, 3,5-difluoro-4-hydroxyphenyl or 2,4-difluoro-3-hydroxyphenyl;

R 8 , R 9 , and R 10 are each independently H, OH, halogen, or alkyl optionally substituted with one or more halogen or OH, or R 8 and R 9 taken together with the carbon atoms to which they are attached form a 3- to 6-membered carbocyclic or heterocyclic ring comprising 1 or 2 heteroatoms selected from N, O, and S, or R 9 and R 10 taken together with the carbon atoms to which they are attached form a 3- to 6-membered carbocyclic or heterocyclic ring comprising 1 or 2 heteroatoms selected from N, O, and S;

m is 0, 1, or 2;

n is 0 or 1; and

p is 0 or 1;

the method comprising the steps of

a) protecting a compound of formula D to prepare a compound of formula D1;

b) reacting the compound of formula D1 with an activating reagent to convert the C-12 alcohol into a leaving group, thus preparing a compound of formula D2;

c) reacting the compound of formula D2 with a base to prepare compound of formula D3;

d) reacting a compound of formula D3 with a hydrogen source in the presence of a hydrogenation catalyst to generate a compound of formula D4; and

e) reacting a compound of formula D4 with deprotecting reagents and a reducing agent to prepare a compound of formula D5,

wherein R 11 is a protecting group, X is —(CHR 8 ) m (CHR 9 ) n (CHR 10 ) p —R 7 , where R 7 , R 8 , R 9 , and R 10 may be protected by R 11 or another protecting group

20 . The method of claim 20 , wherein the compound of formula D5 is compound of formula 45