CN1964939B - Compounds and compositions for delivering active agents - Google Patents

Abstract

The present invention provides compounds and compositions for delivering active agents. The invention also provides methods of their application and preparation.

Description

Compounds and compositions for delivering active agents

This application claims the benefit of the following patent applications: U.S. Provisional Application No. 60/576,088, filed June 1, 2004, U.S. Provisional Application No. 60/576,397, filed June 1, 2004, U.S. Provisional Application No. 60/576,397, filed June 1, 2004 Provisional Application No. 60/576,105, U.S. Provisional Application No. 60/571,090, filed May 14, 2004, U.S. Provisional Application No. 60/571,092, filed May 14, 2004, U.S. Provisional Application No. 60/571,092, filed May 14, 2004 No. 60/571,195, U.S. Provisional Application No. 60/571,194, filed May 14, 2004, U.S. Provisional Application No. 60/571,093, filed May 14, 2004, U.S. Provisional Application No. 60, filed May 14, 2004 /571,055, U.S. Provisional Application No. 60/571,151, filed May 14, 2004, U.S. Provisional Application No. 60/571,315, filed May 14, 2004, U.S. Provisional Application No. 60/571,144, filed May 14, 2004 and US Provisional Application No. 60/571,089, filed May 14, 2004, all of which are incorporated herein by reference.

technical field

The present invention relates to compounds and compositions for the delivery of active agents (eg, biologically or chemically active agents) to a target. These compounds are well suited to form non-covalent mixtures with active agents for oral or other administration to animals. The invention also discloses the preparation and application method of the composition.

Background technique

Conventional means of delivering active agents are often severely limited by biological, chemical and physical barriers. Typically, these barriers are created by the environment in which delivery occurs, the environment of the target for delivery, and/or the target itself. Biologically and chemically active agents are particularly susceptible to this barrier.

This barrier is erected by the body during the delivery of biologically and chemically active pharmacological and therapeutic agents to animals. Examples of physical barriers are skin, epithelium, lipid bilayers, and membranes of various organs, which are relatively impermeable to certain active agents, but must pass through the barrier before reaching a target site, such as the circulatory system. Chemical barriers include, but are not limited to, pH changes and degradative enzymes in the gastrointestinal (GI) tract.

These barriers are of particular importance in the design of oral delivery systems. Oral delivery of many active agents is the route of choice for administration to animals if biological, chemical and physical barriers are not present. Among the variety of substances not typically affected by oral administration include biologically or chemically active peptides, such as calcitonin and insulin; polysaccharides, such as mucopolysaccharides, including but not limited to heparin; heparinoids; antibiotics; and other organic substances . These substances may be rapidly rendered ineffective or destroyed by acid hydrolysis, enzymes, etc. in the gastrointestinal tract. In addition, the size and structure of macromolecular drugs can also inhibit absorption.

Early methods of oral administration of the affected pharmacological substances relied on the co-administration of adjuvants such as resorcinol and nonionic surfactants such as polyoxyethylene oleyl ether and n-hexadecyl polyvinyl ether to artificially increase Permeability of the gut wall, and co-administration of enzyme inhibitors (eg, trypsin inhibitor, diisopropylfluorophosphate (DFP), and aprotinin) to inhibit enzymatic degradation. Liposomes have also been described as drug delivery systems for insulin and heparin. However, wide-scale application of this drug delivery system has been hampered by the following reasons: (1) the system requires toxic amounts of adjuvants or inhibitors; (2) suitable low molecular weight cargoes (i.e., active agents) are not available; ( 3) the system exhibits poor stability and insufficient shelf life; (4) the system is difficult to prepare; (5) the system fails to protect the active agent (cargo); (6) the system adversely alters the active agent; or (7) the system does not allow or facilitate activity drug absorption.

Proteinoid microspheres have been used to deliver drugs. See, eg, US Patent Nos. 5,401,516, 5,443,841 and Re. 35,862. Additionally, some modified amino acids have been used to deliver drugs. See, eg, U.S. Patent Nos. 5,629,020, 5,643,957, 5,766,633, 5,776,888, and 5,866,536, and International Patent Publication Nos. 32130, WO01/32596, WO01/44199, WO01/51454, WO02/02509, WO02/15959, WO02/16309, WO02/20466, WO02/19969, WO02/69937, WO03/45306.

More recently, polymers have been linked to modified amino acids or derivatives thereof via linking groups to provide polymeric delivery agents. The modified polymer may be any polymer, but preferred polymers include, but are not limited to, polyethylene glycol (PEG) and its derivatives. See, eg, International Patent Publication No. WO 00/40203.

However, there remains a need for simple, inexpensive delivery systems that are easy to prepare and can deliver a wide range of active agents through multiple routes.

Contents of the invention

The present invention provides compounds and compositions that facilitate the delivery of active agents. The delivery agent compounds of the present invention include the following compounds and pharmaceutically acceptable salts thereof:

Compound A

in:

R ₁ is -(CH ₂ ) _m -R ₈ , wherein m=0 or 1;

R ₂ -R ₆ are independently selected from hydrogen, hydroxyl, halogen, C ₁ -C ₄ alkyl, C ₂ -C ₄ alkenyl, C ₂ -C ₄ alkynyl, C ₁ -C ₄ alkoxy and cyano base;

R ₇ is selected from C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl and C ₂ -C ₁₀ alkynyl;

_R is selected from cyclopentyl, cyclohexyl and phenyl, wherein when _R is phenyl, m=1; and

R ₈ is optionally substituted with C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, halogen or hydroxy, or combinations thereof.

In one embodiment, R ⁷ is Ci _alkyl .

In another embodiment, R ⁷ is C ₂ alkyl.

In another embodiment, R ⁷ is C ₃ alkyl.

In another embodiment, R ⁷ is C ₄ alkyl.

In another embodiment, R ⁷ is C ₅ alkyl.

In another embodiment, R ⁷ is C ₆ alkyl.

In another embodiment, R ⁷ is C ₇ alkyl.

In another embodiment, R ⁷ is C ₈ alkyl.

Preferred compounds include, but are not limited to, the following compounds and pharmaceutically acceptable salts thereof:

(compound 1)

(compound 2)

(compound 3)

(compound 4)

(compound 5)

       (化合物6)

(Compound 6)

(compound 7)

(Compound 8)

(Compound 9)

 (化合物10)

(compound 10)

(Compound 11)

(compound 12)

(compound 13)

(compound 14)

(化合物15)

(compound 15)

(化合物16)

(compound 16)

(化合物17)

(compound 17)

(compound 18)

(compound 19)

(化合物20)

(compound 20)

(compound 21)

(compound 22)

Other delivery agent compounds of the invention include those of the formula: and pharmaceutically acceptable salts thereof:

(Compound B)

in:

R ₁ is C ₁ -C ₆ alkyl or C ₂ -C ₆ alkenyl,

R ₂ -R ₆ are independently selected from hydrogen, hydroxyl, halogen, C ₁ -C ₄ alkyl, C ₂ -C ₄ alkenyl, C ₂ -C ₄ alkynyl, C ₁ -C ₄ alkoxy and cyano base, and

R ₇ is selected from C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl and C ₂ -C ₁₀ alkynyl.

In one embodiment, R2-R6 are independently hydrogen, methyl, halo, methoxy.

In another embodiment, R2-R6 are independently hydrogen, methyl, chloro, methoxy.

In another embodiment, R2-R6 are independently hydrogen, methyl, fluoro, methoxy.

In another embodiment, R2-R6 are independently hydrogen, methyl, iodo, methoxy.

In another embodiment, R2-R6 are independently hydrogen, methyl, bromo, methoxy.

In another embodiment, R1 is C ₁ -C ₃ alkyl.

In another embodiment, R1 is methyl.

In another embodiment, R1 is ethyl.

In another embodiment, R1 is isopropyl.

In another embodiment, R2 is methyl.

In another embodiment, R2 is halo.

In another embodiment, R2 is chlorine.

In another embodiment, R2 is fluoro.

In another embodiment, R4 is methyl.

In another embodiment, R4 is methoxy.

In another embodiment, R4 is halogen.

In another embodiment, R4 is chlorine.

In another embodiment, R4 is fluoro.

In another embodiment, R4 is cyano.

In another embodiment, R7 is Ci _alkyl .

In another embodiment, R7 is _C2 alkyl.

In another embodiment, R7 is _C2 alkyl with methyl branching.

In another embodiment, R7 is _C3 alkyl.

In another embodiment, R7 is _C3 alkyl with methyl branching.

In another embodiment, R7 is _C4 alkyl.

In another embodiment, R7 is _C5 alkyl.

In another embodiment, R7 is _C6 alkyl.

In another embodiment, R7 is _C7 alkyl.

In another embodiment, R7 is _C8 alkyl.

Preferred compounds include but are not limited to the following compounds and their pharmaceutically acceptable salts

compound

twenty three

               

6-(isopropyl-

Phenyl-aminomethyl

acyl)-caproic acid

compound

twenty four

               

6-[isopropyl

(Phenyl)amine

base]-6-oxo

Caproic acid

compound

25

              

5-[isopropyl

(Phenyl)amine

base]-3-methyl

-5-oxopentanoic acid

compound

26

             

                                                           

base)amino]-3-

Methyl-5-oxo

Valeric Acid

compound

27

                        

...

                                                                      

Oxoheptanoic Acid

compound

28

                        

                                                

                                                                           

base]-5-oxo

Valeric Acid

compound

29

                       

...

                                                                                               ,

base]-4-oxo

Butyric acid

compound

30

                       

8-[methyl(4-methyl

                                                               

base]-8-oxo

Bitter

compound

31

                      

8-[(4-methoxy

                                                                                                                                                                                    ,

Amino]-8-oxy

                               

compound

32

                     

8-[Ethyl(Benzene

                                                              

Oxooctanoic Acid

compound

33

                    

8-[(4-Chlorobenzene

Ethyl)(Ethyl)Ammonia

base]-8-oxo

Bitter

compound

34

            

8-[(4-Fluorobenzene

                                                                                               

base]-8-oxo

Bitter

compound

35

           

                                                    

Methylphenyl) ammonia

base]-10-oxo

Decanoic Acid

compound

36

           

...

                                                                                                   ,

base]-3-oxo

Propionic acid

compound

37

            

  …

Ethyl)(Ethyl)Ammonia

base]-3-oxo

Propionic acid

compound

38

            

                                                

Ethyl)(Ethyl)Ammonia

base]-4-methyl

                                               

compound

39

            

...

                                                                                               ,

base]-10-oxo

Decanoic Acid

compound

40

                  

                                                      

Ammonia

base]-4-oxo

Butyric acid

compound

41

                 

                                              

base) (methyl) ammonia

base]-5-oxo

Valeric Acid

compound

42

                

                                            

                                   

base]-7-oxo

Heptanoic Acid

compound

43

                

                                                               

                                   

base]-6-oxo

Caproic acid

compound

44

                

                                                     

base (4-methylbenzene

                                                                   

                               

compound

45

               

                                                

base) (methyl) ammonia

                          

-5-oxopentanoic acid

compound

46

               

8-[Methyl(2-methyl

                                

base]-8-oxo

Bitter

compound

47

              

                                            

                                                           

                                                           

-5-oxopentanoic acid

compound

48

              

5-[isopropyl

(Phenyl)Ammonia

                              

-5-oxopentanoic acid

compound

49

             

                                              

                                                         

                            

-5-oxopentanoic acid

compound

50

             

...

                                                                                                   ,

                            

-4-Oxobutanoic acid

compound

51

            

                                                     

                                                         

base]-9-oxo

Pelargonic acid

compound

52

           

8-[Ethyl(2-methyl

                                    

base]-8-oxo

Bitter

compound

53

           

8-[isopropyl

(Phenyl)Ammonia

base]-8-oxo

Bitter

compound

54

            

                                                  

                                  

base]-9-oxo

Pelargonic acid

compound

55

                 

                                                     

base)amino]-6-

                                     

compound

56

                 

...

                                                         

base]-7-oxo

Enanthate

compound

57

        

                                                       

                                  

base]-9-oxo

Pelargonic acid

compound

58

         

                                                     

base)amino]-10-

Oxodecanoic Acid

compound

59

          

6-[(4-Chlorobenzene

Ethyl)(Ethyl)Ammonia

base]-6-oxo

Caproic acid

compound

60

          

4-[isopropyl

(Phenyl)amine

base]-4-oxo

Butyric acid

compound

61

        

                                                           

base)amino]-4-

Methyl-5-oxo

Valeric Acid

compound

62

        

                                                            

                                                         

base]-4-oxo

Butyric acid

compound

63

        

                                            

                                                         

base]-5-oxo

Valeric Acid

compound

64

        

                                              

                                                           

base]-5-oxo

Valeric Acid

compound

65

        

5-[isopropyl

(Phenyl)amine

base]-5-oxo

Valeric Acid

compound

66

      

                                                     

base)amino]-4-

Methyl-5-oxo

Valeric Acid

compound

67

      

                                                     

base)amino]-5-

                                   

compound

68

     

...

                                

base]-6-oxo

Caproic acid

compound

69

     

...

                                

base]-6-oxo

Caproic acid

compound

70

    

...

                                

base]-6-oxo

Caproic acid

compound

71

    

...

                              

base]-7-oxo

Heptanoic Acid

compound

72

             

...

                                                       

base]-7-oxo

Enanthate

compound

73

             

...

Ethyl)(Ethyl)Ammonia

base]-5-oxo

Valeric Acid

compound

74

             

...

base) (methyl) ammonia

base]-6-oxo

Caproic acid

compound

75

             

...

base) (methyl) ammonia

base]-7-oxo

Heptanoic Acid

compound

76

            

...

base) (methyl) ammonia

base]-7-oxo

Heptanoic Acid

compound

77

            

...

                                                                                                                                                               ,

Amino]-7-oxo

                                     

compound

78

            

8-[(4-cyanobenzene

base) (methyl) ammonia

base]-8-oxo

Bitter

compound

79

            

8-[(4-Fluorobenzene

                                                                                                   ,

base]-8-oxo

Bitter

compound

80

            

                                            

                                                                                                   ,

base]-9-oxo

Pelargonic acid

compound

81

            

                                                     

                                                                                                   ,

base]-9-oxo

Pelargonic acid

compound

82

            

...

Ethyl)(Ethyl)Ammonia

                                

Heptanoic Acid

compound

83

            

8-[(4-Chlorobenzene

base) (methyl) ammonia

base]-8-oxo

Bitter

compound

84

            

                                                

base) (methyl) ammonia

base]-9-oxo

Pelargonic acid

compound

85

                

9-[isopropyl

(Phenyl)Ammonia

base]-9-oxo

Pelargonic acid

compound

86

                

                                                         

                                  

base]-9-oxo

Pelargonic acid

compound

87

                

...

Methylphenyl) ammonia

base]-10-oxo

Decanoic Acid

compound

88

                

                                                       

(Phenyl)amine

base]-10-oxo

Decanoic Acid

compound

89

                

                                                     

base)amino]-10-

Oxodecanoic Acid

compound

90

                

...

base) (methyl) ammonia

base]-9-oxo

Pelargonic acid

compound

91

        

...

                                                                                                                                                          ,

Amino]-9-oxo

                                                                       

compound

92

        

...

                                                                                                                                                          ,

Amino]-10-oxy

                                     

compound

93

        

...

                                                                          

base)amino]-10-

Oxodecanoic Acid

compound

94

        

...

base) (methyl) ammonia

base]-10-oxo

Decanoic Acid

compound

95

        

...

base) (methyl) ammonia

base]-10-oxo

Decanoic Acid

compound

96

        

...

Ethyl)(Ethyl)Ammonia

base]-9-oxo

Pelargonic acid

compound

97

          

...

base) (methyl) ammonia

base]-10-oxo

Decanoic Acid

compound

98

          

...

Ethyl)(Ethyl)Ammonia

base]-10-oxo

Decanoic Acid

compound

99

          

                                                    

base (4-methylbenzene

base)amino]-4-

Oxobutyric acid

compound

100

          

                                                    

base (2-methylbenzene

base)amino]-4-

Oxobutyric acid

compound

101

          

                                                                     

(Phenyl)ammonia

                                                     

-4-Oxobutanoic acid

compound

102

          

                                                

base (4-methylbenzene

base)amino]-5-

                                 

compound

103

                

                                              

base (2-methylbenzene

base)amino]-5-

                                   

compound

104

               

                                                           

                                                 

                                                     

-4-Oxobutanoic acid

compound

105

              

                                                   

base (4-methylbenzene

                                                                    

                                     

compound

106

             

                                                     

base (2-methylbenzene

                                                                  

                                   

compound

107

              

                                              

base) (methyl) ammonia

                                           

-5-oxopentanoic acid

compound

108

              

                                                      

Ammonia

                                           

-4-Oxobutanoic acid

compound

109

         

                                                       ,

base) (methyl) ammonia

                                             

-5-oxopentanoic acid

compound

110

          

                                                       ,

Ethyl)(Ethyl)Ammonia

                                                         

-5-oxopentanoic acid

Other delivery agent compounds of the invention include those of the formula: and pharmaceutically acceptable salts thereof:

Compound C

n=1 to 9, and

R ₁ to R ₅ are independently hydrogen, C ₁ -C ₄ alkyl, C ₁ -C ₄ alkoxy, C ₂ -C ₄ alkenyl, halogen, hydroxyl, -NH-C(O)-CH ₃ or -OC ₆ H ₅ .

Preferred delivery agent compounds include, but are not limited to, compounds having the following structural formula and salts thereof:

In one embodiment, n=2-8.

In another embodiment, n=8.

In another embodiment, n=7.

In another embodiment, n=6.

In another embodiment, n=5.

In another embodiment, n=4.

In another embodiment, n=3.

In another embodiment, n=2 and the remaining R groups are H.

In another embodiment, n=8 and the remaining R groups are H.

In another embodiment, n=7 and the remaining R groups are H.

In another embodiment, n=6 and the remaining R groups are H.

In another embodiment, n=5 and the remaining R groups are H.

In another embodiment, n=4 and the remaining R groups are H.

In another embodiment, n=3 and the remaining R groups are H.

In another embodiment, n=2 and the remaining R groups are H.

In another embodiment, R1 and R5 are hydrogen.

In another embodiment, R1 and R5 are hydrogen and n=2.

In another embodiment, R3 is hydroxyl.

In another embodiment, R3 is hydroxyl and N=8.

In another embodiment, R1 is hydroxyl.

In another embodiment, R1 is hydroxyl and N=8.

In another embodiment, R3 is methoxy.

In another embodiment, R3 is methoxy and N=2.

In another embodiment, R3 is methoxy and N=3.

In another embodiment, R2 and R4 are halogen and N=2.

In another embodiment, R2 and R4 are fluoro.

In another embodiment, R2 and R4 are fluoro and N=2.

In another embodiment, R1 and R3 are methyl.

In another embodiment, R1 and R3 are methyl and N=2.

In another embodiment, R2 and R4 are methyl, R3 is methoxy and N=4.

In another embodiment, R3 is isopropyl.

In another embodiment, R3 is isopropyl and N=3.

In another embodiment, R1 is methoxy.

In another embodiment, R1 is methoxy and N=2.

In another embodiment, R3 is halogen.

In another embodiment, R3 is halogen and N=2.

In another embodiment, R3 is fluoro and N=2.

In another embodiment, R3 is methoxy.

In another embodiment, R3 is methoxy and N=4.

In another embodiment, R2 and R4 are methyl.

In another embodiment, R2 and R4 are methyl and N=2.

In another embodiment, R2 and R4 are methyl and N=4.

In another embodiment, R2 and R4 are methyl and N=6.

In another embodiment, R2 and R3 are methyl and N=4.

In another embodiment, R2 and R3 are methyl and N=2.

In another embodiment, R1 and R4 are methyl and N=2.

In another embodiment, R1 and R4 are halogen.

In another embodiment, R1 and R4 are halogen and N=2.

In another embodiment, R1 and R4 are halogen and N=4.

In another embodiment, R1 and R4 are chloro.

In another embodiment, R1 and R4 are chloro and N=2.

In another embodiment, R1 and R4 are chloro and N=4.

In another embodiment, R1 and R4 are hydroxyl.

In another embodiment, R1 and R4 are hydroxyl and N=8.

In another embodiment, compounds 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 128, 129, 130, 132, 133, 134, 136 and/or 138 are excluded from compound C .

Preferred compounds include, but are not limited to, the following compounds.

Compound 111

Compound 112

Compound 113

Compound 114

Compound 115

Compound 116

Compound 117

Compound 118

Compound 119

Compound 120

Compound 121

Compound 122

Compound 123

Compound 124

Compound 125

Compound 126

Compound 128

Compound 129

Compound 130

Compound 132

Compound 133

Compound 134

Compound 136

Compound 138

Other delivery agent compounds of the invention include those of the formula: and pharmaceutically acceptable salts thereof:

(Compound D)

R1 to R4 are independently hydrogen, C ₁ -C ₄ alkyl, C ₂ -C ₄ alkenyl, halogen, C ₁ -C ₄ alkoxy or hydroxy.

In one embodiment, R1 and R4 are independently hydrogen, methyl, methoxy, halo or isopropyl.

In one embodiment, R1 to R4 are all hydrogen.

In another embodiment, R2 and R4 are halogen, preferably bromine or preferably chlorine, or preferably iodine, or preferably fluorine.

In another embodiment, R2 and R4 are halogen, preferably bromine or preferably chlorine, or preferably iodine, and R1 and R3 are hydrogen.

In another preferred embodiment, R2 and R4 are isopropyl.

In another preferred embodiment, R2 and R4 are isopropyl, and R1 and R3 are hydrogen.

In another preferred embodiment, R4 is methyl.

In another preferred embodiment, R4 is methyl and R1 and R3 are hydrogen.

In another preferred embodiment, R3 is halogen, preferably chlorine.

In another preferred embodiment, R3 is halogen, preferably chlorine and R1, R2 and R4 are hydrogen.

In another preferred embodiment, R3 is methoxy.

In another preferred embodiment, R3 is methoxy, and R1, R2 and R4 are hydrogen.

In another preferred embodiment R2 is halogen, preferably bromine.

In another preferred embodiment, R2 is halogen, preferably bromine, and R1, R2 and R4 are hydrogen.

In another preferred embodiment, R2 is halogen, preferably chlorine.

In another preferred embodiment, R2 is halogen, preferably chlorine, and R1, R3 and R4 are hydrogen.

In another preferred embodiment, R2 is methoxy.

In another preferred embodiment, R2 is methoxy, and R1, R3 and R4 are hydrogen.

In another preferred embodiment, R2 is methyl.

In another preferred embodiment, R2 is methyl, and R1, R3 and R4 are hydrogen.

Preferred delivery agent compounds include, but are not limited to, compounds having the following structural formula and salts thereof:

Compound 140

                                                  

Compound 141

|

Acid

Compound 142

|

Acid

Compound 143

|

Acid

Compound 144

|

Acid

Compound 145

|

Compound 146

...

Butyric acid

Compound 147

                                                 

Compound 148

...

Compound 149

...

Butyric acid

Compound 150

|

Acid

Compound 151

...

base)-butyric acid

Other delivery agent compounds of the invention include those of the formula: and pharmaceutically acceptable salts thereof:

(Compound E)

in

One of R1 to R5 has the following general structure

-(CH ₂ ) _n -COOH

where n=0 to 6;

The remaining four groups of R1 to R5 are independently hydrogen, _C1 - _C4 alkyl, _C2 - _C4 alkenyl, halogen, _C1 - _C4 alkoxy, or hydroxyl; and

R6 to R10 are independently hydrogen, C ₁ -C ₄ alkyl, C ₂ -C ₄ alkenyl, halogen, C ₁ -C ₄ alkoxy or hydroxy.

In one embodiment, n=0-4.

In another embodiment, n=0.

In another embodiment, n=1.

In another embodiment, R1-R10 are preferably independently hydrogen, halo, methyl and methoxy.

In another embodiment, R1-R10 are preferably independently chloro, halo, methyl and methoxy.

In another embodiment, when the general structure -( _CH2 ) _n -COOH is attached at the R1 position, the remaining R groups are hydrogen.

In another embodiment, when the general structure -( _CH2 ) _n -COOH is attached at the R1 position, the remaining R group is hydrogen and n=0.

In another embodiment, when the general structure -( _CH2 ) _n -COOH is attached at the R1 position, the remaining R group is hydrogen and n=1.

In another embodiment, when the general structure -( _CH2 ) _n -COOH is attached at the R3 position, the remaining R group is hydrogen.

In another embodiment, when the general structure -( _CH2 ) _n -COOH is attached at the R3 position, the remaining R group is hydrogen and n=0.

In another embodiment, when the general structure -( _CH2 ) _n -COOH is attached at the R3 position, the remaining R group is hydrogen and n=1.

In another embodiment, R5 is methoxy when the general structure -( _CH2 ) _n -COOH is attached at the R2 position.

In another embodiment, when the general structure -( _CH2 ) _n -COOH is attached at the R2 position, R5 is methoxy and the remaining R groups are hydrogen.

In another embodiment, R5 is methoxy when the general structure -( _CH2 ) _n -COOH is attached at the R2 position, and n=0.

In another embodiment, when the general structure -( _CH2 ) _n -COOH is attached at the R2 position R5 is methoxy and n=0 and the remaining R groups are hydrogen.

In another embodiment, R1 and R5 are methyl when the general structure -( _CH2 ) _n -COOH is attached at position R3.

In another embodiment, when the general structure -( _CH2 ) _n -COOH is attached at position R3, R1 and R5 are methyl and the remaining R groups are hydrogen.

In another embodiment, R1 and R5 are methyl and n=0 when the general structure -( _CH2 ) _n -COOH is attached at R3.

In another embodiment, when the general structure -( _CH2 ) _n -COOH is attached at position R3, R1 and R5 are methyl and n=0, and the remaining R groups are hydrogen.

In another embodiment, R1 or R5 is methoxy and n=0 when the general structure -( _CH2 ) _n -COOH is attached at R3.

In another embodiment, when the general structure -( _CH2 ) _n -COOH is attached at position R3, R1 or R5 is methoxy and n=0, and the remaining R groups are hydrogen.

In another embodiment, R2 or R4 is halogen, preferably chlorine, when the general structure -( _CH2 ) _n -COOH is attached at the R3 position.

In another embodiment, when the general structure -( _CH2 ) _n -COOH is attached at position R3, R2 or R4 is halogen, preferably chlorine, and the remaining R groups are hydrogen.

In another embodiment, R2 or R4 is halogen, preferably chlorine and n=0 when the general structure -( _CH2 ) _n -COOH is attached at R3.

In another embodiment, when the general structure -( _CH2 ) _n -COOH is attached at the R3 position R2 or R4 is halogen, preferably chlorine and n=0, the remaining R groups are hydrogen.

In one embodiment, compounds 152, 153, 154, 155, 156, 157 and/or 158 are excluded from compound E.

Preferred compounds include, but are not limited to, the following compounds and pharmaceutically acceptable salts thereof:

Compound 152

                                                             

Compound 153

                                                                              

Compound 154

                                                                 

Compound 155

  (4-Benzyloxy-3-methoxy-phenyl)-acetic acid

Compound 156

4-(Benzyloxy)-2-chlorobenzoic acid

Compound 157

                                                         

Compound 158

  (4-Benzyloxy-phenyl)-acetic acid

Compound 159

                                                         

Other delivery agent compounds of the invention include those of the formula: and pharmaceutically acceptable salts thereof:

Compound F

in

n=1 to 9; and

R ₁ to R ₉ are independently hydrogen, C ₁ -C ₄ alkyl, C ₂ -C ₄ alkenyl, halogen, C ₁ -C ₄ alkoxy or hydroxy.

According to a preferred embodiment, n=3 to 7, preferably, in a preferred embodiment, n=3, preferably, in another preferred embodiment, n=4; preferably, in another In a preferred embodiment, n=5; preferably, in another preferred embodiment, n=6; preferably, in another preferred embodiment, n=7.

According to another preferred embodiment, _R1 to _R8 are hydrogen.

According to another preferred embodiment, _R3 is halogen, preferably, in one embodiment, _R3 is chlorine, preferably, in another embodiment, _R3 is bromine.

According to another preferred embodiment, R ₂ is methoxy.

According to another preferred embodiment, R ₂ is methyl.

According to another preferred embodiment, R ₃ is methoxy.

According to another preferred embodiment, R ₃ is methyl.

According to another preferred embodiment, R ₆ is methoxy.

According to another preferred embodiment, _R9 is hydrogen.

According to another preferred embodiment, R ₉ is hydroxyl.

According to another preferred embodiment, R ₉ is halogen, preferably, in one embodiment chlorine.

According to another preferred embodiment, _R3 and _R6 are both methoxy.

According to another preferred embodiment, _R3 and _R6 are both methoxy and the remaining R groups are hydrogen.

According to another preferred embodiment, R ₂ is methyl and R ₃ is chloro.

According to another preferred embodiment, _R2 is methyl and _R3 is chlorine and the remaining R groups are hydrogen.

According to another preferred embodiment, R ₂ is methyl and R ₉ is chloro.

According to another preferred embodiment, R ₂ is methyl and R ₉ is chloro and the remaining R groups are hydrogen.

According to another preferred embodiment, R ₃ is methyl and R ₉ is chloro.

According to another preferred embodiment, _R3 is methyl and _R9 is chlorine and the remaining R groups are hydrogen.

Preferred delivery agent compounds include, but are not limited to, compounds having the following structural formula and salts thereof:

Compound 160

6-(2-(2-Hydroxybenzoyl)phenoxy)hexanoic acid

Compound 161

Sodium 8-(2-(2-hydroxybenzoyl)phenoxy)octanoate

Compound 162

5-(2-(2-Hydroxybenzoyl)-4-methoxyphenoxy)pentanoic acid

Compound 163

5-(2-(2-Hydroxybenzoyl)phenoxy)pentanoic acid

Compound 164

5-(2-(2-Hydroxy-5-methoxybenzoyl)-4-methoxyphenoxy)pentanoic acid

Compound 165

4-(2-(2-Hydroxybenzoyl)-5-methoxyphenoxy)butanoic acid

Compound 166

4-(2-(2-Hydroxybenzoyl)phenoxy)butanoic acid

Compound 167

4-(2-Chlorobenzoyl-4-methylphenoxy)butanoic acid

Compound 168

4-(2-Benzoyl-5-methoxyphenoxy)butanoic acid

Compound 169

4-(2-Benzoyl-4-chlorophenoxy)butanoic acid

Compound 170

4-(2-Benzoyl-4-bromophenoxy)butanoic acid

Compound 171

4-(2-(2-Chlorobenzoyl-5-methylphenoxy)butanoic acid

Compound 172

4-(2-(2-Chlorobenzoyl-4-methylphenoxy)butanoic acid

Compound 173

4-(2-Benzoyl-4-chloro-5-methylphenoxy)butanoic acid

Compound 174

3-(5-Chloro-2-hydroxybenzamido)propionic acid

Other delivery agent compounds of the invention include those of the formula: and pharmaceutically acceptable salts thereof:

(Compound G)

in

R1 to R5 are independently hydrogen, C ₁ -C ₄ alkyl, C ₂ -C ₄ alkenyl, halogen, C ₁ -C ₄ alkoxy, hydroxyl or -O-(CH ₂ ) _n -COOH (where n to 12);

At least one of _R1 to _R5 has the following general structure

-O-(CH ₂ ) _n -COOH

where n=1 to 12; and

R6 to R10 are independently hydrogen, C ₁ -C ₄ alkyl, C ₂ -C ₄ alkenyl, halogen, C ₁ -C ₄ alkoxy or hydroxy.

Preferably, only one of R1 to R5 has the formula -O-( _CH2 ) _n -COOH. In other words, four of R1 to R5 are independently hydrogen, C ₁ -C ₄ alkyl, C ₂ -C ₄ alkenyl, halogen, C ₁ -C ₄ alkoxy, or hydroxyl, and R1 to R5 The remaining one group in is -O-(CH ₂ ) _n -COOH (where n is 1 to 12).

In a preferred embodiment n=1 to 12.

In another preferred embodiment n=1 to 10.

In another preferred embodiment n=1 to 6.

In another preferred embodiment n=1 to 4.

In another preferred embodiment n=10.

In another preferred embodiment n=4.

In another preferred embodiment n=1.

When the general structure -( _CH2 ) _n -COOH is attached at the R1 position, all other R groups are hydrogen.

When the general structure -( _CH2 ) _n -COOH is attached at the R1 position, all other R groups are hydrogen and n=3.

When the general structure -( _CH2 ) _n -COOH is attached at the R3 position, all other R groups are hydrogen.

When the general structure -( _CH2 ) _n -COOH is attached at the R3 position, all other R groups are hydrogen and n=1.

When the general structure -( _CH2 ) _n -COOH is attached at the R3 position, all other R groups are hydrogen and n=4.

When the general structure -( _CH2 ) _n -COOH is attached at the R3 position, all other R groups are hydrogen and n=10.

Preferred compounds include, but are not limited to, the following compounds and pharmaceutically acceptable salts thereof:

Compound 175

                                                               

Compound 176

  (4-Benzyloxy-phenoxy)-acetic acid

Compound 177

11- (2-cymbal oxygenzen oxygen) eleven

Acid

Compound 178

                                                                 ,

Mixtures of these delivery agent compounds may also be used.

The invention also provides compositions comprising a delivery agent compound of the invention and at least one active agent. These compositions deliver the active agent into the biological system of choice with increased or improved bioavailability of the active agent compared to administration of the active agent without the delivery agent compound.

The invention also provides dosage unit forms comprising the compositions. Dosage units may be in liquid or solid form, such as tablets, capsules or granules, including powders or sachets.

Another embodiment is a method of administering an active agent to an animal by administering to the animal a composition comprising at least one delivery agent compound of the present invention and the active agent. Routes of administration include oral, intracolonic and pulmonary routes.

Another embodiment is a method of treating a disease or obtaining a desired physiological effect in an animal by administering a composition of the invention.

Another embodiment is the administration of a composition of the invention to an animal that would benefit from the composition and/or to an animal in need of the active agent.

Another embodiment is a method of preparing a composition of the invention by admixing at least one delivery agent compound of the invention and at least one active agent.

Detailed description of the invention

definition

As used herein and in the appended claims, the singular forms include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a molecule" includes one or more such molecules, "a reagent" includes one or more of the different reagents, and reference to "the method" or "the method" includes references to those of ordinary skill in the art. Equivalent steps and methods are well known to the person, and these methods may be improvements or substitutions of the methods described herein.

The term "polymorphs" refers to crystallographically distinct forms of a substance.

The term "hydrate" as used herein includes, but is not limited to, (i) substances comprising molecularly bound water and (ii) crystalline substances comprising one or more molecules of water of crystallization or crystalline materials comprising free water.

The term "solvate" as used herein includes, but is not limited to, molecular or ionic complexes of molecules or ions of a solvent with molecules or ions of a delivery agent.

The term "delivery agent" refers to any delivery agent compound disclosed herein or incorporated by reference herein, including pharmaceutically acceptable salts thereof.

An "effective amount of a pharmaceutical composition" is an amount of a pharmaceutical composition effective to treat or prevent a disease in a subject to which it is administered over a period of time, eg, to provide a therapeutic effect within the intended dosage interval.

The term "treating" refers to prophylactically preventing, curing, curing, alleviating, alleviating, altering, correcting, ameliorating, ameliorating or affecting a condition (eg, a disease), a symptom of a condition, or a tendency to develop a condition.

An "effective amount of delivery agent" is an amount of delivery agent that facilitates absorption of a desired amount of active agent.

The term "subject" includes mammals such as rodents, cows, pigs, dogs, cats, primates and especially humans.

The term "AUC" as used herein means the area under the plasma concentration-time curve calculated by the trapezoidal rule over complete dosage intervals, eg 24 hour intervals.

When the term "average" precedes a pharmacokinetic value (eg mean peak value), it represents the arithmetic mean of the pharmacokinetic value, unless otherwise stated.

As used herein, the term "about" means within 10%, preferably within 5%, more preferably within 1% of the specified value. Additionally, the term "about" means a value that may fall within a scientifically acceptable range of error for that type of value, depending on the degree of qualitative nature of the approach given by available tools.

"Indication" means the use of an administered drug to prevent or treat a disease, and can be used interchangeably with "treatment".

The term "substituted" as used herein includes, but is not limited to, substitution with any one or any combination of the following substituents: halogen, hydroxy, C ₁ -C ₄ alkyl, and C ₁ -C ₄ alkoxy.

The terms "alkyl", "alkoxy", "alkenyl", "alkenylene", "alkyl(arylene)" and "aryl(alkenyl)" respectively include, but are not limited to Linear or branched alkyl, alkoxy, alkenyl, alkenylene, alkyl (arylene) and aryl (alkenyl).

"Peptide YY" or "PYY" refers to a Peptide YY polypeptide obtained or derived from any species. Thus, the term "PYY" includes human full-length, 36 amino acid peptides as described in International Publication No. WO02/47712 (which is the PCT text of U.S. Patent Publication No. 2002/0141985, which is incorporated herein by reference) SEQ ID NO: 2 and Tatemoto, Proc Natl Acad Sci U.S.A. 79:2514-8, 1982, and species variations of PYY including, for example, murine, hamster, chicken, cow, rat, and dog PYY. "PYY agonist" means any compound that causes the effect of PYY to reduce the availability of nutrients, for example a compound having the following properties (1) in food intake, gastric emptying, pancreatic secretion or weight loss assays (in WO02/47712 and U.S. Pat. described in Examples 1, 2, 5, or 6 of Publication No. 2002/0141985) and (2) in the Y receptor assay (Example 10 of WO02/47712 and US Patent Publication No. 2002/0141985) or competition In a sexual binding assay, it specifically binds to labeled PYY or PYY [3-36] from some tissues with abundant Y receptors, including, for example, the region postrema (Example 9 of WO 02/47712 and U.S. Patent Publication No. 2002/0141985 ), wherein the PYY agonist is not a pancreatic polypeptide. Preferably, the PYY agonist has a binding affinity in this assay of greater than about 1 μM, more preferably an affinity of greater than about 1 nM to about 5 nM.

The agonists may include polypeptides or chemical or small molecules with functional PYY regions, PYY active fragments. PYY agonists may be peptidic or non-peptidic compounds, and include "PYY agonist analogs," which refer to any compound that is structurally similar to PYY, typically due to interaction with PYY receptors or other receptors. Or the PYY receptor itself may have PYY activity by binding to or otherwise interacting directly or indirectly with a receptor with which it interacts to elicit a biological response. The compounds include PYY derivatives, PYY fragments, extended PYY molecules with more than 36 amino acids, truncated PYY molecules with less than 36 amino acids, and substituted PYY molecules with one or more different amino acids, or the above Any combination of molecules. The compounds may also be modified by methods such as pegylation, amidation, glycosylation, acylation, sulfation, phosphorylation, acetylation and cyclization.

One of the PYY agonist analogs is PYY[3-36], identified as SEQ ID NO:3 of WO02/47712 and U.S. Patent Publication No. 2002/0141985; Eberlein, Eysselein et al., Peptides 10:797-803 (1989) and Grandy, Schimiczek et al., Regul Pept 51:151-9 (1994). Polypeptides with numbers in brackets refer to truncated polypeptides having the full-length peptide sequence including the amino acid positions in brackets. Thus, PYY[3-36] has the same sequence as amino acids 3 to 36 of PYY. PYY[3-36] comprised about 40% of the total peptide YY-like immune activity in human and canine intestinal extracts and about 36% of the total plasma peptide YY immune activity in the fasted state, while slightly less after meals. higher than 50%. It is apparently a dipeptidyl peptidase-IV (DPP4) cleavage product of peptide YY. Peptide YY [3-36] is reported to be a selective ligand for Y2 and Y5 receptors, which is pharmacologically unique among the preferred N-terminal truncated (i.e., its C-terminal fragment) neuropeptide Y analogs sex. PYY agonists can bind to PYY receptors with higher and lower affinity, exhibit longer or shorter half-lives in vivo or in vitro, or be more or less effective than native PYY.

Other suitable PYY agonists include those PYY agonists described in International Publication No. WO98/20885, which is incorporated herein by reference.

The term "heparin" as used herein refers to all forms of heparin, including but not limited to unfractionated heparin, heparanoid, dermatan, chondroitin, low molecular weight heparin (such as tinzaparin (including tinzaparin) Sodium)), very low molecular weight heparin and ultra low molecular weight heparin. Non-limiting examples include unfractionated heparins, such as sodium heparin (eg, sodium heparin USP, available from Scientific Protein Labs of Waunakee, WI). Heparin typically has a molecular weight of about 1,000 or 5,000 to about 30,000 Daltons. The term "low molecular weight heparin" generally refers to heparin in which at least about 80% by weight of the heparin has a molecular weight between about 3000 and about 9000 Daltons. Non-limiting examples of low molecular weight heparins include tinzaparin, enoxaprin, and daltiparin. Tinzaparin has been approved by the US Food & Drug Administration for the treatment of acute lower extremity venous thrombosis with or without pulmonary embolism when used in combination with warfarin sodium. The sodium salt of Pharmion Corporation ^(TM) tinzaparin is available under the trademark Innohep of Boulder, CO. The term "very low molecular weight heparin" generally refers to heparin in which at least about 80% by weight of the heparin has a molecular weight between about 1500 and about 5000 Daltons. A non-limiting example of a very low molecular weight heparin is bemiparin. The term "ultra-low molecular weight heparin" generally refers to heparin in which at least about 80% by weight of the heparin has a molecular weight between about 1000 and about 2000 Daltons. A non-limiting example of ultra-low molecular weight heparin is fondiparinux.

Delivery agent

The delivery agents of the invention may be in free acid or pharmaceutically acceptable salt form. Suitable pharmaceutically acceptable salts include, but are not limited to, organic and inorganic salts such as alkali metal salts such as sodium, potassium and lithium; alkaline earth metal salts such as magnesium, calcium or barium; ammonium salts; basic amino acids such as lysine amino acid or arginine; and organic amines such as dimethylamine or pyridine. In one embodiment, the salt is the sodium salt. Salts may be monovalent or polyvalent, such as monosodium and disodium salts. Salts may also be solvates, including ethanol solvates, and hydrates. Non-limiting examples of pharmaceutically acceptable salts include salts with sodium, hydrochloric acid, sulfuric acid, phosphoric acid, citric acid, acetic acid, sulfate, phosphate, chloride, bromide, iodide, acetate, propionic acid salt, hydrobromic acid, sodium hydroxide, potassium hydroxide, ammonium hydroxide and potassium carbonate. These salts can be prepared by methods well known in the art. For example the sodium salt can be prepared by dissolving the transfer agent in ethanol and adding aqueous sodium hydroxide. The delivery agent can be purified by recrystallization or in separate or sequential fractions on one or more solid phase chromatographic supports. Suitable solvent systems for recrystallization include, but are not limited to, acetonitrile, methanol, and tetrahydrofuran. Fractionation can be performed (i) on a suitable chromatographic support such as alumina using a methanol/n-propanol mixture as the mobile phase, (ii) by reverse phase chromatography using a trifluoroacetic acid/acetonitrile mixture as the mobile phase phase, or (iii) by ion exchange chromatography using water or an appropriate buffer as the mobile phase. When performing anion exchange chromatography, it is preferred to use a 0-500 mM sodium chloride gradient.

The delivery agent may comprise a polymer attached to a linking group selected from the group consisting of -NHC(O)NH-, -C(O)NH-, -NHC(O), -OOC-, -COO-, - NHC(O)O-, -OC(O)NH-, -CH ₂ NH-NHCH ₂ -, -CH ₂ NHC(O)O-, -OC(O)NHCH ₂ -, -CH ₂ NHCOCH ₂ O- , -OCH ₂ C(O)NHCH ₂ -, -NHC(O)CH ₂ O-, -OCH ₂ C(O)NH-, -NH-, -O-, and carbon-carbon bonds, provided that the polymer transfers The agent is not a polypeptide or polyamino acid. The polymer can be any polymer, including but not limited to alternating copolymers, block copolymers and random copolymers, which is safe for use in mammals. Preferred polymers include, but are not limited to, polyethylene, polyacrylate, polymethacrylate, poly(oxyethylene), poly(propylene), polypropylene glycol, polyethylene glycol (PEG), and derivatives of these polymers and combinations of these polymers. The molecular weight of the polymer typically ranges from about 100 to about 200,000 Daltons. The molecular weight of the polymer preferably ranges from about 200 to about 10,000 Daltons. In one embodiment, the molecular weight of the polymer ranges from about 200 to about 600 Daltons and more preferably ranges from about 300 to about 550 Daltons. US Patent No. 6,627,228 is incorporated herein by reference in its entirety.

The amount of delivery agent in the solid pharmaceutical composition is a delivery agent effective amount and can be determined for a particular composition according to those methods well known in the art. Typically, the weight ratio of delivery agent to active agent ranges from about 0.1:1 to about 1000:1 and preferably from about 1:1 to about 300:1. The weight ratio will vary depending on the active agent and the particular indication for which the active agent is administered.

对于本发明其他适合的传递剂在以下专利中描述：美国专利号6,846,844、6,699,467、6,693,208、6,693,073、6,663,898、6,663,887、6,646,162、6,642,411、6,627,228、6,623,731、6,610,329、6,558,706、6,525,020、6,461,643、6,461,545、6,440,929、6,428,780 、6,413,550、6,399,798、6,395,774、6,391,303、6,384,278、6,375,983、6,358,504、6,346,242、6,344,213、6,3.31,318、6,313,088、6,245,359、6,242,495、6,221,367、6,180,140、5,541,155、5,693,338、5,976,569、5,643,957、5,955,503、6,100,298、5,650,386、5,866,536 , 5,965,121, 5,989,539, 6,001,347, 6,071,510 and 5,820,881.本发明的传递剂也在以下专利中描述：美国专利申请公开号20050009748、20040110839、20040106825、20040068013、20040062773、20040022856、20030235612、20030232085、20030225300、20030198658、20030133953、20030078302、20030072740、20030045579、20030012817、20030008900、20020155993 . The delivery agents of the present invention are also described in the following patents: International Publication Nos. WO2005/020925, WO2004/104018, WO2004/080401, WO2004/062587, WO2003/057650, WO2003/057170, WO2003/045331, WO2003/045306, WO2003/02 , WO2002/100338, WO2002/070438, WO2002/069937, WO02/20466, WO02/19969, WO02/16309, WO02/15959, WO02/02509, WO01/92206, WO01/70219, WO01/51441, WO991/4 /34114, WO01/32596, WO01/32130, WO00/07979, WO00/59863, WO00/50386, WO00/47188, WO00/40203 and WO96/30036. Each of the US patents and US and International published patent applications listed above is incorporated herein by reference. These delivery agents can be prepared by methods well known in the art, such as those described in the aforementioned patents and published patent applications. For example, SNAC can be prepared by methods known in the art, such as those described in US Patent Nos. 5,650,386 and 5,866,536, and US Patent Application Publication No. 2002/0065255.

active agent

Active agents suitable for use in the present invention include biologically active agents and chemically active agents including, but not limited to, pesticides, pharmaceutical agents, and therapeutic agents. Suitable active agents include those that are reduced, ineffective or destroyed by acid hydrolysis, enzymatic action, etc. in the gastrointestinal tract. Also included as suitable active agents are macromolecular substances whose physicochemical properties (including size, structure or charge) inhibit or prevent absorption when administered orally.

For example, substances that enter the body or could benefit from improved pharmacokinetics including delivery, such as when oral bioavailability is limited or non-existent. These substances are biologically or chemically active agents suitable for use in the present invention and include, but are not limited to, macromolecules such as peptides, including proteins or polypeptides, including dipeptides; hormones; and carbohydrates, including monosaccharides, polysaccharides, including disaccharides , mixtures of mucopolysaccharides; carbohydrates; lipids; and small polar organic molecules (i.e., polar organic molecules having a molecular weight of 500 Daltons or less); nucleosides, other organic compounds; Oral bioavailability or specific compounds with limited oral bioavailability, including those that do not per se (or pass only a fraction of the administered dose) the gastrointestinal mucosa and/or are susceptible to chemical cleavage by acids and enzymes in the gastrointestinal tract compounds; or any combination of these compounds.

Additional examples include, but are not limited to, the following, including their synthetic, natural or recombinant origin:

Amylin and amylin agonists; Adrenocorticotropic hormone;

antigen; Antimicrobials, including antibiotics, antibacterials, and antifungals; non-limiting examples of antibiotics include antibiotics acting on Gram-positive bacteria, bactericidal antibiotics, lipopeptide antibiotics, and cyclic peptide antibiotics, such as daptomycin and its analogues; Anti-migraine drugs such as BIBM-4096BS and other calcitonin gene-related protein antagonists, sumatriptan succinate; Antiviral drugs, including acyclovir and valacyclovir; Atrial natriuretic factor; Bisphosphonates, including alendronate, clodronate, etidronate, ibandronate, incadronate ), minodronate, neridronate, olpadronate, pamidronate, risedronate, tiludronate tiludronate, zoledronate, EB1053 and YH529; Calcitonin, including salmon calcitonin, eel calcitonin, porcine calcitonin and human calcitonin; Cholecystokinin (CCK) and CCK agonists including CCK-8; Sodium cromolyn (sodium cromolyn or disodium cromolyn); cyclosporine; Deferoxamine (DFO); Erythropoietin; Exendins and exendin agonists, including exendin-3, exendin-4; Filgrastim (filgrastim); Follicle-stimulating hormone (recombinant and natural follicle-stimulating hormone); Glucagon-like peptide 1 (GLP-1), glucagon and glucagon-like peptide 2 (GLP-2); Glucocerebrosidase; Gonadotropin-releasing hormone; growth hormone releasing factor; growth hormone releasing hormone;

Included are secretagogues, analogs, fragments, mimetics or polyethylene glycol (PEG) modified derivatives of these compounds; or any combination of these substances.

delivery system

Compositions of the invention include one or more delivery agent compounds of the invention, and one or more active agents. In one embodiment, one or more delivery agent compounds, or salts of these compounds, or polyaminoacids or peptides of which compounds or salts form one or more units, may be formed by mixing with the active agent prior to administration to form The administration composition acts as a delivery agent.

The composition for administration may be in liquid form. Solution media can be water (eg for salmon calcitonin, parathyroid hormone and erythropoietin), 25% propylene glycol in water (eg for heparin) and phosphate buffer (eg for recombinant human growth hormone). Other pharmaceutical excipients include polyethylene glycol. Drug solutions can be prepared by mixing a solution of the delivery agent compound with a solution of the active agent just prior to administration. Alternatively, a solution of the delivery agent compound (or active agent) may be mixed with a solid form of the active agent (or delivery agent compound). The delivery agent compound and active agent can also be mixed as a dry powder. The delivery agent compound and the active agent may also be mixed during manufacture.

The drug solution may optionally include additives such as phosphate buffered saline, citric acid, glycol or other dispersing agents. Stabilizing additives may be incorporated into the solution, preferably in a concentration range of between about 0.1% and 20% (w/v).

In addition, the composition for administration may be in the form of a solid, such as a tablet, capsule or granules, such as a powder or sachet. Solid dosage forms can be prepared by admixing the compound in solid form with the active agent in solid form. Alternatively, solids can be obtained from solutions of the compound and active agent by methods well known in the art, such as freeze-drying (lyophilization), precipitation, crystallization, and solid dispersion.

The administration compositions of the invention may also comprise one or more enzyme inhibitors. The enzyme inhibitors include but are not limited to compounds such as actinin or epiactinin and their derivatives. Other enzyme inhibitors include, but are not limited to, aprotinin (Trasylol) and Bowman-Birk inhibitors.

The amount of active agent employed in the compositions for administration of the invention is that amount of the particular active agent that is effective for the intended indication. The amount of active agent in the composition is typically a pharmacologically, biologically, therapeutically or chemically effective amount. However, when the composition is used in a dosage unit form, its amount may be lower than that stated above, since the dosage unit form may include multiple delivery agent compound/active agent combinations or may include separate pharmacological, biological, therapeutic or chemical agents. effective amount. The total effective amount can then be administered in cumulative units comprising the total effective amount of active agent.

The total amount of active agent employed can be determined by those methods well known in the art. However, because the compositions of the present invention can deliver the active agent more effectively than compositions comprising only the active agent, lower levels of biologically or chemically active agents can be administered to patients than those employed in previous dosage unit forms or delivery systems. amount of biologically or chemically active agent while still achieving the same blood level and/or therapeutic effect.

The presently disclosed delivery agent compounds facilitate the delivery of biologically and chemically active agents, particularly oral, intranasal, sublingual, gastric, intestinal, including intraduodenal, jejunal and ileal delivery, subcutaneous, buccal, intracolonic, rectal , vaginal, mucosal, pulmonary, transdermal, intradermal, parenteral, intravenous, intramuscular, and ocular systems, and across the blood-brain barrier.

Dosage unit forms may also contain one or a combination of the following: excipients, diluents, disintegrants, lubricants, plasticizers, coloring agents, flavoring agents, taste-masking agents, sugars, sweeteners, salts And pharmaceutical excipients, including but not limited to water, 1,2-propanediol, ethanol, olive oil or any combination of these excipients.

The compounds and compositions of the present invention are useful for administering biologically or chemically active agents to any animal including, but not limited to, birds such as chickens; mammals such as rodents, cows, pigs, dogs, cats, primates animals, especially humans; and insects.

The system is particularly advantageous for the delivery of chemical or biological agents that encounter environmental damage or exhibit lower efficacy before the active agent reaches the target area (i.e., the area from which the active agent of the delivery composition is released) and in the animal to which the active agent is administered. active agent. In particular, the compounds and compositions of the invention are useful for orally administered active agents, especially those that are not normally orally deliverable, or those for which improved delivery is desired.

Compositions comprising a compound and an active agent have the effect of delivering the active agent to a selected biological system and of increasing or improving the bioavailability of the active agent compound relative to administration of the active agent without the delivery agent. Delivery can be improved by delivering more of the active agent over a period of time, or by delivering the active agent for a specific period of time (e.g., accelerated action or delayed delivery), or by delivering the active agent at a specific time or over a period of time (e.g., sustained transmission).

Another embodiment of the present invention is a method of treating or preventing a disease or obtaining a desired physiological effect, such as those listed in the table below, in an animal by administering a composition of the present invention. Preferably, the composition is administered in an effective amount for treating or preventing the desired disease or obtaining the desired physiological effect. Specific indications for active agents can be found in the following literature: (1) the Physicians' Desk Reference (58th Ed., 2004, Medical Economics Company, Inc., Montvale, NJ) and (2) Fauci, AS, et al. ., Harrison's Principles of Internal Medicine (14th Ed., 1998, McGraw-Hill Health Professions Division, New York), both of which are incorporated herein by reference. The active agents in the table below include their analogs, fragments, mimetics and polyethylene glycol modified derivatives.

active agent disease or physiological effect Amylin and amylin agonists; obesity Adrenocorticotropic hormone; High cholesterol (to lower cholesterol) antigen; Infect Antiviral drugs, including acyclovir, valacyclovir; Viral infections, including herpes simplex types I and II Growth hormone (including human recombinant growth hormone and growth hormone releasing factor and its analogues) growth disorder Interferons, including alpha, beta, and gamma Viral infections, including chronic cancer, hepatitis, and multiple sclerosis Interleukins (eg, interleukin-1, interleukin-2, interleukin-11, interleukin-21) Viral infection; cancer; cell-mediated immunity and transplant rejection; Insulin; Insulin-like Growth Factor IGF-1 diabetes heparin Treatment and prevention of thrombosis, including (deep vein thrombosis); prevention of blood clotting Calcitonin, including salmon calcitonin, eel calcitonin, porcine calcitonin and human calcitonin; Osteoporosis; Bone Disorders; Bone Pain; Pain Relief (including Pain Associated with Osteoporosis or Cancer) Cholecystokinin (CCK) and CCK agonists including CCK-8; obesity Erythropoietin Anemia; HIV/HIV-Treatment-Associated Anemia; Chemotherapy-Induced Anemia atrial natriuretic factor Vasodilation antigen Infect

CPHPC Reduces amyloid deposition and systemic amyloidosis often (but not always) associated with Alzheimer's disease, type 2 diabetes, and other amyloid-based diseases Monoclonal antibodies (antibodies include Retuxin, TNF-α soluble receptor;) Prevention of graft rejection; cancer; used in assays to detect disease Leptin (OB protein) obesity Somatostatin/octreotide Bleeding ulcers; erosive gastritis; varicose veins; diarrhea; acromegaly; adrenotropic pituitary adenomas; secretory pancreatic tumors; carcinoid syndrome; reduced herniation/thyroid-associated ophthalmopathy; reduced macule Edema/Retinopathy Protease inhibitor HIV infection/AIDS ACTH High cholesterol (to lower cholesterol) GnRH Ovulatory disturbance (stimulation of ovulation) Oxytocin Dysfunction of labor (stimulated contractions) Luteinizing Hormone Releasing Hormone; Follicle Stimulating Hormone Regulate reproductive function Glucocerebrosidase Gaucher disease (metabolic lipoprotein) Thrombopoietin Thrombocytopenia Filgrastim (granulocyte colony-stimulating factor); GM-CSF, (recombinant granulocyte-macrophage colony-stimulating factor) Shorten the duration of chemotherapy-induced neutropenia and thereby treat or prevent infection in chemotherapy patients; inhibit the growth of Mycobacterium Intracellular Avium or kill Mycobacterium Intracellular Avium infection (MAC) prostaglandins hypertension Cyclosporine transplant rejection

Vasopressin Nocturnal Enuresis; Antidiuretics sodium cromolyn Asthma; Allergy Vancomycin Treatment or prophylaxis of antimicrobial-induced infections, including but not limited to methicillin-resistant Staphalococcus aureus and Staph. epidermiditis gallium nitrate Osteoporosis; Paget's disease; Inhibits osteoclasts; Promotes osteoblast activity, hypercalcemia, including cancer-associated hypercalcemia, urethral (urinary tract) malignancies; Antineoplastic, cancer, including urethra and bladder Carcinoma; Lymphoma; Malignancy (including bladder cancer); Leukemia; Management of bone metastases (and associated pain); Multiple myeloma; Impaired immune response, including allograft rejection; Interference with iron metabolism; Promotion of cell migration; Trauma Repair; impair or treat the infectious process of mycobacteria, including but not limited to Mycobacterium Tubercolosis and Mycobacterium Avium complex Deferoxamine (DFO) iron overload Parathyroid hormone (PTH), including fragments thereof. osteoporosis; bone disease Antimicrobials Infections, including but not limited to Gram-positive infections vitamins Treat and prevent vitamin deficiencies

Bisphosphonates, including alendronate, clodronate, etidronate, ibandronate, incadronate, minodronate, neridronate, Padronate, pamidronate, risedronate, tiludronate, zoledronate, EB1053 and YH529; Osteoporosis; Paget's disease; osteoma and metastases (and associated pain); breast cancer; including as adjuvant therapy in early breast cancer; management of bone metastases (and associated pain), including in relation to breast, prostate, and lung cancers bone metastases; inhibition of osteoclasts; promotion of osteoblast activity; treatment and/or prevention of bone mineral density (bmd) loss; multiple myeloma; prevention of bone complications associated with malignant osteolysis; fibrous dysplasia ; pediatric osteogenesis imperfecta; hypercalcemia, urethral (urinary tract) malignancy; reflex sympathetic dysbehavioral syndrome, acute back pain after comminuted vertebral fracture, chronic inflammatory joint disease, renal bone disease, extrosseous calcification, pain relief, vitamin D intoxication, periarticular ossification Anti-migraine drugs, such as BIBM-4096BS, BIBN4096BS-(1-piperidinecarboxamide.N-[2-[[5-amino-1-[[4-(4-pyridyl)-1-piperazinyl] Carbonyl]pentyl]amino]-1-[(3,5-dibromo-4-hydroxyphenyl)methyl]-2-oxoethyl]-4-(1,4-dihydro-2-oxo Dai-3(2H-quinazolinyl)-[R-(R ^* , S ^* )]-) and other calcitonin gene-related protein antagonists, sumatriptan succinate; Antimigraine; Calcitonin Gene-Related Peptide Antagonist

Glucagon Improving glycemic control (e.g., treatment of hypoglycemia and control of hypoglycemic reactions), obesity; diagnostic aid in radioactivity detection of stomach, duodenum, small intestine, and colon; therapeutic cardiovascular drugs including, but not limited to, calcium channel blockers, Acute poisoning by beta blockers GLP-1, Exendin-3, Exendin-4 Diabetes mellitus; improving glycemic control (eg, treating hypoglycemia and controlling hypoglycemic reactions), obesity Dipeptidyl peptidase IV (DPP-4) inhibitors Diabetes mellitus; improving glycemic control (eg, treating hypoglycemia), obesity Vaccines, including those against anthrax or Yersinia pestis, influenza, and herpes. prevent or minimize disease or infection Peptide YY (PYY) and PYY-like peptides Obesity, diabetes, eating disorders, insulin resistance syndrome

For example, one embodiment of the invention is a method of treating a patient suffering from or susceptible to diabetes mellitus by administering insulin of the pharmaceutical formulation of the invention. Other active agents, including those listed in the table above, may be used in combination with the pharmaceutical formulations of the present invention.

Following administration, the active agent present in the composition or dosage unit form is absorbed into the circulation. The bioavailability of a substance can be readily assessed by measuring known pharmacological activities in blood, such as heparin-induced increase in blood clotting time or calcitonin-induced decrease in circulating calcium levels. Alternatively, circulating levels of the active agent itself can be directly determined.

additive

The solid pharmaceutical compositions and unit dosage forms of the present invention may include other active agents and pharmaceutically acceptable additives, such as excipients, carriers, diluents, stabilizers, plasticizers, binders, glidants, disintegrants , bulking agents, lubricants, plasticizers, colorants, film-forming agents, flavoring agents, taste-masking agents, sugars, sweeteners, preservatives, pharmaceutical excipients, surfactants, and any combination of any of the above substances. Preferably, these additives are pharmaceutically acceptable additives such as those described in Remington's, The Science and Practice of Pharmacy, (Gennaro, AR, ed., 20 ^th edition, 2003, Mack Pub. Co.) , which is incorporated herein by reference.

Suitable binders include, but are not limited to, starch, gelatin, sugars (such as sucrose, syrup, and lactose), dibasic calcium phosphate, natural and synthetic gums (such as acacia), sodium alginate, carboxymethylcellulose, Methylcellulose, polyvinylpyrrolidone, polyethylene glycol, ethylcellulose and paraffin.

Suitable glidants include, but are not limited to, talc and silicon dioxide (eg, fumed silicon dioxide and colloidal silicon dioxide).

Suitable disintegrants include, but are not limited to, starch, sodium starch glycolate, croscarmellose sodium, crospovidone, clays, celluloses such as purified cellulose, methylcellulose, and carboxymethylcellulose. sodium cellulose), alginates, pregelatinized cornstarch, and gums (such as agar, guar, carob, karaya, pectin, and tragacanth). A preferred disintegrant is sodium starch glycolate.

Suitable bulking agents include, but are not limited to, starch (eg, rice starch), microcrystalline cellulose, lactose (eg, lactose monohydrate), sucrose, dextrose, mannitol, calcium sulfate, dicalcium sulfate, and tricalcium sulfate.

Suitable lubricants include, but are not limited to, stearic acid, stearates (such as calcium stearate and magnesium stearate), talc, boric acid, sodium benzoate, sodium acetate, sodium fumarate, sodium chloride, Polyethylene glycol, hydrogenated cottonseed oil and castor oil.

Suitable surfactants include, but are not limited to, sodium lauryl sulfate, hydroxylated soy lecithin, polysorbate, and propylene oxide and ethylene oxide block copolymers.

dosage form

Solid pharmaceutical compositions of the invention comprising an active agent and a delivery agent may be formulated as solid unit dosage forms. Dosage forms can be, for example, tablets, sachets or capsules, for example hard or soft gelatin capsules. The dosage form can provide rapid, sustained or controlled release of the delivery agent, heparin, and optionally the additional active agent.

The solid pharmaceutical compositions and solid unit dosage forms of the present invention can be prepared as follows. The solid form of the delivery agent is processed (eg, comminuted by passing through a 35 mesh sieve) to provide a powder with a relatively small and preferably uniform particle size. The delivery agent is then mixed with the delivery agent and optionally fillers and/or wetting agents using, for example, a V-blender or smaller apparatus to provide a powder mixture.

The humectant mixture is prepared separately by mixing the humectant, heparin and delivery agent. The mixture may also, for example, comprise water. The formulation of the wetting mixture is chosen so as to wet the heparin when mixed with the aforementioned powder mixture. According to a preferred embodiment, the wetting agent mixture can also be formulated such that it partially dissolves the delivery agent when mixed with the powder mixture.

With continuous mixing, the powder mixture is added in small increments to the wetting agent mixture. Mixing is continued for a sufficient time (eg, 15 minutes) after all of the powder mixture has been added to obtain a homogeneous composition. The resulting composition is typically semi-solid, gel or liquid.

The composition can then be formulated into dosage forms, such as capsules, by methods well known in the art. According to a preferred embodiment, the resulting composition is packaged in soft or hard gelatin capsules (eg Licap Capsugel size 0 hard gelatin capsules). Other suitable methods are described in U.S. Patent Nos. 6,605,298, 6,458,383, 6,261,601, 5,714,477, and 3,510,561; U.S. Patent Application Publication Nos. 2003/0077303 and 2001/0024658; and International Publication No. WO88/10117, all of which are incorporated herein by reference .

Example

The following examples illustrate the invention without limiting it. All percentages are by weight unless otherwise indicated.

分光计(Bruker-Physik AG，Silberstreifen，GERMANY)或400MHz JEOL分光计(JEOL USA，Inc.，Peabody，MA)，应用二甲亚砜(DMSO-d6)作为溶剂进行的。Unless otherwise stated, proton nuclear magnetic resonance (1H NMR) analysis of the compounds listed below was performed at 300 MHz

Spectrometer (Bruker-Physik AG, Silberstreifen, GERMANY) or 400 MHz JEOL spectrometer (JEOL USA, Inc., Peabody, MA) was performed using dimethylsulfoxide (DMSO-d6) as solvent.

Liquid chromatography/mass spectrometry (LC-MS) analyzes were performed with an Agilent Technologies (Palo Alto, California), LC/MSD 1100 (single quadrupole) with the following parameters:

Mobile phase A: 50:950:5 acetonitrile:water:acetic acid (v/v/v).

Mobile phase B: 950:50:5 acetonitrile:water:acetic acid (v/v/v).

Gradient elution: 4 minutes linear gradient 0-100% B; total time per injection is 11 minutes.

Injection volume: 5 μL.

Chromatographic column: ZORBAX Rapid Resolution Cartridge, SB-C18, 2.1×30mm, 3.5μm.

Particle Size, Catalog #873700-902.

Column temperature: 40°C.

UV detection at 244nm.

MSD parameters:

Source: API-ES, positive scan parameters:

Quality range: 125.00-600.00

Collision dissociation voltage: 60V

Gain: 1.0EMV

Threshold: 150

Spray chamber:

Gas temperature: 350deg.D

Drying gas: 12.0L/min

Nozzle pressure: 40psig

Capillary voltage: 4000V positive/negative

The preparation of embodiment 1 compound 1-22

Compounds 1-22 were prepared according to the method of US Patent No. 6,384,278, which is incorporated herein by reference in its entirety.

In xylene, the appropriate N-substituted aniline is mixed with the appropriate dicarboxylic acid monoester and heated in the presence of a boronic acid catalyst. The intermediate amide is hydrolyzed to obtain the final product.

The preparation of embodiment 2 compound 23-34 and 59

A dry 200 mL 3-neck round bottom flask was fitted with a Teflon-coated magnetic stir bar and a vacuum-jacketed Dean-Stark trap topped with a reflux fitting for the nitrogen inlet tube condenser. A reaction vessel was charged with N-isopropyl-N-phenylamine (8.11 g, 60 mmol), boronic acid (0.93 g, 15 mmol) and xylene (88 mL). A portion of 7-ethoxy-7-oxoheptanoic acid (11.29 g, 60 mmol) was added to the stirred reaction mixture. The reaction was heated to reflux using a heating mantle. The azeotropic water began to separate and was collected in the Dean-Stark trap. After reflux for 16 hours, water was collected and the reaction was allowed to cool to ambient temperature to react. The reaction mixture was diluted with ethyl acetate (100 mL), and washed with 2N aqueous HCl (50 mL), and then with saturated sodium bicarbonate solution (60 mL). Most of the organic solvent was removed under vacuum. To the residue was added 2N aqueous sodium hydroxide solution (60 mL). The mixture was heated at 60°C for 4 hours. After cooling to room temperature, the mixture was washed with 60 mL of ethyl acetate. After careful separation from the organic layer, the aqueous phase was evaporated to remove any remaining ethyl acetate. Ice was added to the aqueous solution, followed by aqueous HCl (2N, 60 mL), resulting in the precipitation of a white solid. Stirring was continued for an additional 30 minutes, then the precipitate was collected on a fritted funnel. The collected white solid was successfully washed with water and hexane, then placed under vacuum at room temperature for 12 hours to afford 7.49 g (45%) of 7-[isopropyl(phenyl)amino]-7-oxoheptanoic acid, It is a white solid. HPLC: singlet at 4.83 min; Mp: 62-63°C. ¹ H NMR (DMSO-d6,) δ: 0.95-0.97 (d, 6H), 1.08-1.10 (m, 2H), 1.34-1.40 (m, 4H), 1.76-1.79 (m, 2H), 2.09-2.13 (m, 2H), 4.81-4.85 (m, 1H), 7.18-7.20 (m, 2H), 7.44-7.46 (m, 3H). Quality (M+1): 278. Anal. Calcd. for _C16H23NO3 : C, 69.29; _H , 8.36; N, _5.05 . Found: C, 69.06; H, 8.45; N, 4.99.

Compounds 24-34 and 59 were prepared using the same procedures using appropriate starting materials.

Compound (24)

HPLC: Single peak at 4.43 min. Quality (M+1): 264. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ: 0.95 (d, 6H), 1.30 (m, 2H), 1.40 (m, 2H), 1.80 (m, 2H), 2.00 (m, 2H), 4.80 (m, 1H), 7.15(m, 2H), 7.40(m, 3H). ¹³ C NMR (100 MHz, DMSO-d ₆ ) δ: 21.0, 24.0, 24.5, 33.0, 34.0, 45.0, 128.0, 129.0, 130.0, 138.5, 170.5, 174.0.

Compound (25)

HPLC: Single peak at 4.62 min. Quality (M+1): 264. ¹ H NMR (400MHz, DMSO-d ₆ ) δ: 0.78 (d, 3H), 0.94-0.95 (d, 6H), 1.70-1.72 (m, 1H), 1.80-1.92 (m, 2H), 2.08-2.15 (m, 1H), 2.20-2.30 (m, 1H), 4.75-4.90 (m, 1H), 7.10-7.20 (m, 2H), 7.35-7.50 (m, 3H). ¹³ C NMR (100 MHz, DMSO-d ₆ ) δ: 19.5, 21.0, 27.0, 40.5, 41.0, 45.0, 128.0, 129.0, 130.5, 138.5, 170.0, 174.0.

Compound (26)

HPLC: Single peak at 4.19 min. Quality (M+1): 250. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ: 0.65 (d, 3H), 0.84-0.86 (t, 3H), 1.80-1.90 (m, 3H), 2.01-2.12 (m, 2H), 3.49-3.53 (q, 2H), 7.09-7.11 (d, 2H), 7.20-7.25 (m, 1H), 7.30-7.32 (m, 2H). ¹³ C NMR (100 MHz, DMSO-d ₆ ) δ: 9.18, 15.87, 17.30, 23.35, 39.50, 123.98, 124.72, 125.92, 138.39, 166.17, 168.27, 169.80.

Compound (27)

HPLC: Single peak at 3.92 min. Quality (M+1): 250. ¹ H NMR (400MHz, DMSO-d ₆ ) δ: 1.13(m, 2H), 1.37-1.46(m, 4H), 1.99(m, 2H), 2.10-2.15(t, 2H), 3.15(s, 3H) ), 7.29-7.37 (m, 3H), 7.42-7.47 (m, 2H).

Compound (28)

HPLC: Single peak at 3.72 min. Quality (M+1): 236. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ: 0.79-0.81 (d, 3H), 1.93-2.02 (m, 3H), 2.16-2.30 (m, 2H), 3.15 (s, 3H), 7.27- 7.37 (m, 3H), 7.43-7.48 (m, 2H).

Compound (29)

HPLC: Single peak at 3.88 min. Quality (M+1): 242. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ: 2.21 (m, 2H), 2.49 (m, 2H), 3.13 (s, 3H), 7.37 (m, 2H), 7.58 (m, 2H), 12.10 (br., 1H).

¹³ C NMR (100MHZ, DMSO-d ₆ ) δ: 28.81, 29.0, 36.5, 129.32, 129.58, 132.0, 142.66, 170.58, 173.63.

Compound (30)

HPLC: Single peak at 4.82 min. Quality (M+1): 278. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ: 1.02 (m, 4H), 1.32 (m, 4H), 1.86 (m, 2H), 2.05 (m, 2H), 2.21 (s, 3H), 3.00 ( s, 3H), 7.00 (m, 2H), 7.12 (m, 2H), 11.85 (br., 1H).

Compound (31)

HPLC: Single peak at 4.44 min. Quality (M+1): 294. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ: 1.10 (m, 4H), 1.39 (m, 4H), 1.93 (m, 2H), 2.11 (m, 2H), 3.07 (s, 3H), 3.75 (s, 3H), 6.96 (m, 2H), 7.20 (m, 2H), 11.93 (br., 1H).

Compound (32)

HPLC: Single peak at 4.81 min. Quality (M+1): 278. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ: 0.97(t, 3H), 1.10(m, 4H), 1.39(m, 4H), 1.90(m, 2H), 2.13(m, 2H), 3.58 -3.63 (q, 2H), 7.09-7.24 (d, 2H), 7.34 (m, 1H), 7.41-7.45 (m, 2H).

Compound (33)

HPLC: Single peak at 5.48 min. Quality (M+1): 312. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ: 0.96(t, 3H), 1.10(m, 4H), 1.40(m, 4H), 1.91(m, 2H), 2.12(m, 2H), 3.60 (q, 2H), 7.27 (d, 2H), 7.46 (m, 2H), 11.93 (br., IH).

Compound (34)

HPLC: Single peak at 4.52 min. Quality (M+1): 282. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ: 1.09 (m, 4H), 1.39 (m, 4H), 1.93 (m, 2H), 2.10-2.14 (m, 2H), 3.09 (s.3H) , 3.75 (s 3H), 7.19 (m, 2H), 7.30 (m, 2H), 11.91 (br., 1H).

Compound (59)

HPLC: Single peak at 4.71 min. Quality (M+1): 284. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ: 0.90(t, 3H), 1.35-1.37(m, 4H), 1.87(t, 2H), 2.04(t, 2H), 3.52-3.57(q, 2H), 7.25 (m, 2H), 7.43 (m, 2H), 11.94 (s, 1H).

The preparation of embodiment 3 compound 111-139

(Compound 111) 4-oxo-4-phenyl-butyric acid:

10 g (56 mmol) of 3-benzoylpropionic acid (purchased from Sigma-Aldrich Co., St. Louis, MO) was added to 10 mL of water. The mixture was stirred and 28 mL of 2N sodium hydroxide (aq) was added. The resulting solution was stirred for 2 hours and the solid product was collected after the solution was lyophilized. ¹ H NMR (d6-DMSO): δ7.9, d, 2H, (aryl H); δ7.6, t, 1H, (aryl H); δ7.5, t, 2H, (aryl H) ; δ3.1, t, 2H (CH ₂ at carbonyl alpha position); δ2.2, t, 2H (CH ₂ at COOH alpha position); COOH peak was not observed due to the presence of water in the sample.

(Compound 113) 10-(4-hydroxy-phenyl)-10-oxo-decanoic acid:

Under inert gas, a 500 mL flask equipped with a reflux condenser was charged with sebacic acid (20 g, 296 mmol) and acetic anhydride (280 mL, 2.96 mol). The mixture was heated to reflux for 5 hours. Acetic acid and excess acetic anhydride were removed under reduced pressure. The product was used without further purification.

Under inert gas, to a 500 mL flask equipped with a mechanical stirrer was added oxanedecane-2,11-dione (20 g, 108.5 mmol), phenol (10.22 g, 108.5 mmol) and 200 mL of carbon disulfide. Aluminum (III) chloride (72.34 g, 542 mmol) was added and the reaction was stirred for 72 hours. The carbon disulfide was decanted off, and ice was carefully added until most of the mixture was dissolved. The insoluble material was collected by suction filtration and washed with 2 x 100 mL of water. The solid was then dissolved in 100 mL of 1M aqueous sodium hydroxide solution and acidified with 1M aqueous hydrochloric acid until pH = 7.5. The solid formed was removed by filtration and the acidification of the mother liquor was continued until pH = 2.5. The crude precipitate was collected by filtration and washed with 1 x 100 mL of water. The crude product was dissolved in 100 mL of 1M aqueous sodium hydroxide, then carefully acidified with 1M aqueous hydrochloric acid until pH = 7.5 and the precipitated impurities were removed by filtration. The mother liquor was further acidified until pH2. The crude product was collected by filtration and washed with 2 x 50 mL of water. The product was recrystallized from acetone. The isolated product (1.2 g, 4%) was collected by filtration. Found values: C 69.00, H 7.81%; Theoretical values for C ₁₆ H ₂₂ O ₄ are C: 69.04, H: 7.97%. ¹ H NMR (d6-DMSO): δ12.0, bs, 1H (COOH); δ10.3, bs, 1H (aryl-hydroxyl); δ7.8d, 2H (aryl H); δ6.8, d , 2H, (aryl H); δ2.9, t, 2H (CH ₂ at carbonyl α position); δ2.2, t, 2H (CH ₂ at COOH α position); δ1.5, multiplet, 4H (carbonyl β CH ₂ and COOH β CH ₂ ), δ 1.3, multiplet, 8H (remaining CH ₂ ).

(Compound 114) 10-(2-hydroxy-phenyl)-10-oxo-decanoic acid:

To a 100 mL flask was added dichloromethane (50 mL), 9-bromononanol (7.63 g, 34.2. mmol) and trimethylsilyl chloride (4.5 mL, 35.5 mmol) and allowed to stir under nitrogen for 20 minutes. Triethylamine (5.0 mL, 35.9 mmol) was then added and the resulting reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was then diluted with 80 mL of hexane, filtered, and concentrated under reduced pressure. The resulting residue was diluted again with 80 mL of hexane, filtered, and concentrated under reduced pressure to obtain 9.7 g (96%) of a yellow liquid which was used without further purification.

Under an inert gas, 5.69 g (19.3 mmol) of (9-bromo-nonyloxy)-trimethyl-silane was added dropwise to a 50 mL flask containing metallic magnesium (0.59 g, 24.3 mmol), 20 mL of tetrahydrofuran and a small Block iodine crystals are used to start the Grignard reaction. In a 100 mL flask under inert gas, a solution of salicylaldehyde (2.1 mL, 19.7 mmol) in 20 mL of tetrahydrofuran was cooled with an external ice bath. The cooled aldehyde solution was then treated with 1.0 M lithium bis(trimethylsilyl)amide (20.0 mL, 20 mmol). After stirring for 1 hour, the Grignard reaction was cooled with an external ice bath. The cooled Grignard was then cannulated dropwise into the aldehyde solution over 5 minutes under constant stirring. The resulting reaction mixture was allowed to warm to room temperature and stirring was continued overnight. The reaction was poured into 40 mL of ethyl acetate and treated with 15 mL of saturated aqueous sodium bicarbonate. The organic layer was separated and washed with 2 x 25 mL portions of 4% aqueous hydrochloric acid followed by 1 x 20 mL portion of brine. The organic layer was dried over sodium sulfate, filtered and the solvent was removed under reduced pressure. Residual salicylaldehyde was removed by Kugelrohr distillation and the resulting residue was used without further purification.

重铬酸盐(32.9g，87.5mmol)。(加入过程轻度发热)。将反应混合物在室温下搅拌过夜。将反应混合物倾倒至50mL乙酸乙酯中并且用200mL水、30mL 4％盐酸水溶液、30mL水以及最终的30mL盐水洗涤。然后有机层与10g硅胶一起搅拌15分钟，用硫酸钠干燥，过滤并且在减压下除去溶剂。将类白色粗产物从乙醇/水中重结晶。将产物(0.5g，10％)分离，为类白色固体，mp85-87℃。燃烧分析：实测值：C69.01，H8.36％；C₁₆H₂₂O₄理论值为C：69.54，H：8.02％。¹H NMR(d6-DMSO)：δ12.0，s，1H(COOH)；δ7.9 dd，1H(芳基H)；δ7.5，dt，1H，(芳基H)；δ6.9，复杂的多重峰，2H(芳基H)，3.1，t，2H(羰基α位的CH₂)；δ2.2，t，2H(COOHα位的CH₂)；δ1.6，多重峰，2H(羰基β位的CH₂)，δ1.5，多重峰，2H(COOHβ位的CH₂)，δ1.3，多重峰，8H(其余的CH₂)。A 100 mL flask was charged with 1-(2-hydroxy-phenyl)-undecane-1,11-diol (5.0 g, 18.9 mmol) and 50 mL of dimethylformamide. Add pyridine to it

Dichromate (32.9 g, 87.5 mmol). (Slight heating during addition). The reaction mixture was stirred overnight at room temperature. The reaction mixture was poured into 50 mL of ethyl acetate and washed with 200 mL of water, 30 mL of 4% aqueous hydrochloric acid, 30 mL of water and finally 30 mL of brine. The organic layer was then stirred with 10 g of silica gel for 15 minutes, dried over sodium sulfate, filtered and the solvent was removed under reduced pressure. The off-white crude product was recrystallized from ethanol/water. The product (0.5 g, 10%) was isolated as an off-white solid, mp 85-87°C. Combustion analysis: measured value: _C69.01 , H8.36%; theoretical value _{of C16H22O4} : C: 69.54, H: 8.02%. ¹ H NMR (d6-DMSO): δ12.0, s, 1H (COOH); δ7.9 dd, 1H (aryl H); δ7.5, dt, 1H, (aryl H); δ6.9, Complex multiplet, 2H (aryl H), 3.1, t, 2H (CH ₂ at carbonyl α position); δ2.2, t, 2H (CH ₂ at COOH α position); δ1.6, multiplet, 2H ( CH ₂ at carbonyl β position), δ 1.5, multiplet, 2H (CH ₂ at COOH β position), δ 1.3, multiplet, 8H (remaining CH ₂ ).

(Compound 115) 4-(4-methoxy-phenyl)-4-oxo-butanoic acid:

Under inert gas (nitrogen), 5.25mL (48.3mmol) anisole, 4.83g (48.3mmol) succinic anhydride, 125mL 1,1,2,2- tetrachloroethane and 125 mL nitrobenzene. The reaction vessel was cooled with an external ice bath and stirred for 30 minutes. Aluminum trichloride (14.2 g, 106.4 mmol) was added to the pale yellow solution, which then turned dark reddish brown. The ice bath was removed, and the reaction was stirred at room temperature for 36 hours. The reaction was cooled again with an external ice bath. An acidic solution was prepared by pouring 1 N hydrogen chloride solution into a 100 mL beaker filled with ice. This solution was carefully added to the reaction mixture, initially dropwise until the reaction became clear with a white precipitate. After this point, a 10 mL portion of the glacial/acid mixture was carefully added to check for reaction, followed by the remainder of the glacial/acid mixture. A second 100 mL portion of ice/acid mixture was added, the external ice bath was removed and the white cream was stirred for 2 hours. The precipitate in the emulsion was collected by suction filtration. The solid was dissolved in 300 mL 0.3M sodium hydroxide, washed with 100 mL ethyl acetate, and acidified to ~pH1 with 1M hydrochloric acid. The white precipitate collected by vacuum filtration was washed with 3 x 100 mL deionized water and dried. The product (4.7 g, 47%) was isolated as a white solid, mp 149-150°C. Combustion analysis _: measured value: C63.52, H5.78%; theoretical _value of _C11H12O4 : C: 63.45, H: 5.81%. ¹ H NMR (d6-DMSO): δ12.2, s, 1H (COOH); δ7.9d, 2H (aryl H); δ7.0, d, 2H, (aryl H); δ3.8, s , 3H (H of OMe); δ3.2, t, 2H (CH ₂ at carbonyl α position); δ2.5, t, 2H (CH ₂ at COOH α position).

(Compound 116) 5-(4-methoxy-phenyl)-5-oxo-pentanoic acid:

Compound 116 was prepared in a similar manner to compound 115, except using glutaric anhydride instead of succinic anhydride, mp 141-142°C. Measured values: C64.65 _, H6.34%; theoretical values _{for C12H14O4} : C: 64.85, H: 6.35%. ¹ H NMR (d6-DMSO): δ12.2, s, 1H (COOH); δ7.9d, 2H (aryl H); δ7.0, d, 2H, (aryl H); δ3.8, s , 3H (H of OMe); δ3.0, t, 2H (CH ₂ at carbonyl α position); δ2.3, t, 2H (CH ₂ at COOH α position); δ1.8 quintet, 2H (two CH ₂ between CH ₂ ).

Compound 117 was purchased from Aldrich (St. Louis, MO), Cat. No. 514683.

Compound 118 was purchased from Aldrich (St. Louis, MO), catalog number B12687.

Compound 119 was purchased from Aldrich (St. Louis, MO), catalog number S346810.

Compound 120 was purchased from Reike, Aldrich (St. Louis, MO), Cat. No. 7013D.

Compound 121 was purchased from Reike, Aldrich (St.Louis, MO), Cat. No. 7148C and

(Compound 121) 5-(4-isopropyl-phenyl)-5-oxo-pentanoic acid sodium salt:

In a 250 mL flask, 5-(4-isopropyl-phenyl)-5-oxo-pentanoic acid (5 g, 21.3 mmol) was dissolved in 75 mL of ethanol. Sodium hydroxide (0.85 g, 21.3 mmol) was added and the reaction was stirred overnight under reduced pressure on a rotary evaporator. The solid was dried under vacuum and used without further purification. The measured value is C60.24, _H6.66 , Na9.21%; the theoretical value of _{C14H17O3Na is} C: 61.28, H: 6.98, Na8.38%.

¹ H NMR (D2O): δ7.7, d, 2H (aryl H); δ7.2 d, 2H (aryl H); δ2.9, t, 2H (CH ₂ at carbonyl alpha position); δ2. 8, multiplet, 1H, (CH of isopropyl group); δ2.1, t, 2H (CH ₂ of COOH α position); δ1.8, q, 2H (CH ₂ of carbonyl and COOH β position), δ1. 1,d, 6H ( _CH3 of isopropyl).

Compound 122 was purchased from Aldrich (St. Louis, MO), catalog number B13802.

Compound 123 was purchased from Reike, Aldrich (St. Louis, MO), Cat. No. 7060B.

Compound 124 was purchased from Fischer-Scientific (Hampton, NH), Acros, catalog number 17.522.62.

Compound 125 was purchased from Reike, Aldrich (St. Louis, MO), Cat. No. 7011D.

Compound 126 was purchased from Reike, Aldrich (St. Louis, MO), Cat. No. 7036B.

Compound 128 was purchased from Reike, Aldrich (St. Louis, MO), Cat. No. 7012D.

Compound 129 was purchased from Reike, Aldrich (St. Louis, MO), Cat. No. 7012B.

Compound 130 was purchased from Reike, Aldrich (St. Louis, MO), Cat. No. 7055B.

Compound 132 was purchased from Reike, Aldrich (St. Louis, MO), Cat. No. 7005b.

Compound 133 was purchased from Reike, Aldrich (St. Louis, MO), Cat. No. 7036F.

Compound 134 was purchased from Reike, Aldrich (St. Louis, MO), Cat. No. 7144D.

Compound 136 was purchased from Reike, Aldrich (St. Louis, MO), Cat. No. 7144B.

Compound 138 was purchased from Reike, Aldrich (St. Louis, MO), Cat. No. 7036D.

(Compound 139) 10-(2,5-dihydroxy-phenyl)-10-oxo-decanoic acid:

Under inert gas, a 500 mL flask equipped with a reflux condenser was charged with sebacic acid (20 g, 296 mmol) and acetic anhydride (280 mL, 2.96 mol). The mixture was heated to reflux for 5 hours. Acetic acid and excess acetic anhydride were removed under reduced pressure. The product was used without further purification.

Under inert gas, add the previously prepared oxanedecane-2,11-dione (37.95 g, 206 mmol), 1,4-diacetoxy-benzene (20 g , 103mmol) and 200mL carbon disulfide. Aluminum (III) chloride (68.7 g, 515 mmol) was added and the reaction was stirred for 72 hours. The carbon disulfide was decanted off, and ice was added carefully until most of the mixture was dissolved. The insoluble material was collected by suction filtration and washed with 2 x 100 mL of water. The solid was then dissolved in 50 mL of 1M aqueous sodium hydroxide and stirred for 1 hour. The solution was acidified with 1M aqueous hydrochloric acid until pH=2. The crude product precipitate was collected by filtration and redissolved in acetonitrile (50 mL) and dichloromethane (15 mL) and allowed to slowly precipitate over 1 week. The resulting brown powder was collected by filtration and recrystallized from 10:3 acetic acid:water. The product (0.8 g, 3%) was isolated by filtration. Measured value: C65.55, H7.69%; theoretical value _{of C16H22O5} : C: 65.29, H _: 7.53%. ¹ H NMR (d6-DMSO): δ12.0, s, 1H (COOH); δ11.4, s, 1H (aryl-hydroxyl); δ9.2, s, 1H (aryl-hydroxyl); δ7. 2 d, 1H (aryl H); δ7.0, dd, 1H, (aryl H); δ6.8, d, 1H (aryl H), 3.0, t, 2H (CH ₂ at carbonyl alpha position) ; δ2.2, t, 2H (CH ₂ at COOH α position); δ1.6, multiplet, 2H (CH ₂ at carbonyl β position), δ1.5, multiplet, 2H (CH ₂ at COOH β position), δ 1.3, multiplet, 8H (rest _CH2 ).

The preparation of embodiment 4-compound 140-151

In general, compounds are prepared according to a four-step process. First, the appropriate substituted salicylic acid and ethyl 3-aminobutyrate were mixed with dichloroethane (EDC)/hydroxybenzotriazole (HOBt)/dichloromethane (DCM). In the second step, basic ion exchange resin A-15/A-21 (available from Rohm and Haas, Philadelphia, PA) was added. In the third step, after partial work-up, the product is reacted with potassium trimethylsiliconate (KOTMS)/tetrahydrofuran (THF). In the fourth step, IRC-50 resin (Rhohm & Haas, Philadelphia, PA) was added.

Add salicylic acid (4.57 mmol), DCM (10 mL), EDC (1.05 g, 5.48 mmol), HOBt (838 mg, 5.48 mmol), DMF (2 mL), and ethyl-3-aminobutyric acid to each scintillation vial Ester (600 mg, 4.57 mmol). All vials were tightly capped, placed on a J-Kem reaction block (J-Kem Scientific Inc., St. Lois, MO), shaken and heated (150 rpm, 35° C.) overnight. All reactants had one significant spot based on TLC. Amberlyst-21 and Amberlyst-15 resins (approximately 2.5 g, 11 mmol) were added to each vial and shaking continued overnight at ambient temperature. The reactions were filtered, the resin was washed with DCM (2 x 5 mL), and the collected filtrates from each reaction were pooled into a new scintillation vial. The filtrate was blown under a stream of nitrogen to a volume of about 2 mL.

To each vial was added a 1.2 M solution of potassium trimethylsiliconate (KOTMS) in THF (10 mL, 12 mmol). More THF was added to some reactions if necessary to obtain a shakeable slurry. All vials were tightly capped, placed on a J-Kem reaction block, shaken and heated (150 rpm, 60°C, 6 hours). The reaction mass was cooled and IRC-50 resin (3 g, 30 mmol) was added to each vial to treat the potassium salt. Add DCM if necessary to make the resin suspendable and easy to shake. The reaction was shaken overnight. The reactions were filtered, the resin was washed with DCM (2 x 5 mL) and DMF if necessary to dissolve the solids, and the collected filtrates from each reaction were combined in new, tared scintillation vials. At this point a small portion of the filtrate was removed and diluted with 1:1 ACN/ _H2O for LC-MS. The filtrate was blown dry under nitrogen flow. To remove traces of DMF, the vial was placed in a vacuum oven at 50°C.

Some reaction mixtures contained considerable amounts of esters based on LC-MS analysis. These materials were treated again with KOTMS. To each vial was added 1.2M KOTMS in THF (8 mL, 9.6 mmol). All vials were tightly capped, placed in a Pierce reaction block, stirred and heated (60°C, 5 hours). The reaction mass was cooled and IRC-50 resin (2 g, 20 mmol) was added to each vial to treat the potassium salt. DCM was added if necessary to make the resin suspendable and easy to stir. The reaction was stirred over the weekend. The reaction was filtered through a silica gel packed column, the resin and silica gel were washed with DCM (1 x 5 mL), then 2:5 MeOH/DCM (3 x 7 mL) and the filtrate collected from each reaction was combined in a new, tared scintillation vial middle. At this point a small portion of the filtrate was removed and diluted with 1:1 ACN/ _H2O for LC-MS. The filtrate was blown dry under nitrogen flow.

All other reaction mixtures from the first KOTMS treatment were taken up in 10:1 DCM/MeOH and filtered through a plug of silica gel, eluting with more 10:1 DCM/MeOH. At this point a small portion of the filtrate was removed and diluted with 1:1 ACN/ _H2O for LC-MS. The filtrate was blown dry under nitrogen flow.

Selective Preparation of Compounds 140-151

Add 3,5-diisopropylsalicylic acid (25.0 g, 112.5 mmol), HOBt (20.6 g, 135.0 mmol), ethyl-3-aminobutyrate (18.0 g, 123.7 mmol) and dioxane (400 mL). The resulting mixture was stirred at ambient temperature. A portion of EDC (25.9 g, 135.0 mmol) was added and stirring continued overnight. HPLC of the reaction mixture at this point showed HOBt, perhaps traces of starting salicylic acid, and a new major product. Another portion of EDC (5 g, 26.0 mmol) was added and stirring continued overnight. Another HPLC showed essentially no change. The reaction was quenched with water (400 mL) and the dioxane was removed using a rotary evaporator. The resulting oil/water mixture was poured into a IL separatory funnel and DCM (400 mL) was added. A large amount of white solid formed. EtOAc was added to attempt to separate the two layers without success. The separatory funnel was drained and the organics were removed on a rotary evaporator. The water/oil mixture was extracted with EtOAc (500 mL, then 200 mL). The combined EtOAc layers were washed with aqueous HCl (10%, 2 x 200 mL), aqueous NaOH (10%, 2 x 200 mL) and brine (50 mL, then 200 mL). The organics were dried over _Na2SO4 and removed on rotavap to afford _a brown oil with a small amount of white solid. HPLC analysis indicated that the white solid was residual HOBt and the brown oil was the expected product. The brown oil was aspirated from the flask, avoiding as much as possible of the white solid. The brown oil was taken up in EtOAc (500 mL ₎ , washed with NaOH (10%, 2 x 200 mL) and dried over _Na2SO4 . EtOAc was removed on the rotavap to afford a brown oil. HPLC at this point showed one major peak and no HOBt.

The viscous oil was dissolved in THF (200 mL) and KOTMS (31.7 g, 247.4 mmol) was added. The resulting viscous mixture was stirred overnight. HPLC indicated completion of the reaction as one peak. IRC-50 resin (37 g, 370 mmol, 1.5 eq.) and 100 mL DCM were added to suspend the resin, then stirred for several hours. Filtered, washed the resin with DCM (3 x 50 mL) and concentrated on a rotary evaporator to give a brown oil. Recrystallization from ACN/acetone was attempted without success. Based on solubility at this point it is clear that the material is primarily a potassium salt. The oil was taken in _H2O /ACN, heated until clear, filtered hot, and cooled to ambient temperature. The filtrate was treated with aqueous HCl and the resulting solid precipitate was separated and ground into powder to obtain E1528: 9.13g, HPLC retention time was 6.7 minutes 100%, KF 0.47, NMR was consistent with the structure, elemental analysis theoretical value was C: 66.11, H : 8.21, N: 4.54, Found C: 65.62, H: 8.19, N: 4.46.

Compound No. and Name Molecular Weight MS(M+H) Pure LC-MS

Degree Approximate Percentage

Compound 140 223.2306 224 83

3-(2-Hydroxy-benzamido)-butyric acid

Compound 141 381.0326 382 71

3-(3,5-Dibromo-2-hydroxy-benzamido)-butyric acid

Compound 142 292.1206 292 77

3-(3,5-Dichloro-2-hydroxy-benzamido)-butyric acid

Compound 143 475.0234 476 82

3-(2-Hydroxy-3,5-diiodo-benzamido)-butyric acid

Compound 144 237.2577 238 75

3-(2-Hydroxy-3-methyl-benzamido)-butyric acid

Compound 145 257.6756 258 82

3-(4-Chloro-2-hydroxy-benzamido)-butyric acid

Compound 146 253.2571 254 75

3-(2-Hydroxy-4-methoxy-benzamido)-butyric acid

Compound 147 302.1316 303 82

3-(5-Bromo-2-hydroxy-benzamido)-butyric acid

Compound 148 257.6756 258 78

3-(5-Chloro-2-hydroxy-benzamido)-butyric acid

Compound 149 253.2571 254 77

3-(2-Hydroxy-5-methoxy-benzamido)-butyric acid

Compound 150 237.2577 238 82

3-(2-Hydroxy-5-methyl-benzamido)-butyric acid

Compound 151 307.3931 308 89

3-(2-Hydroxy-3,5-diisopropyl-benzamido)-butyric acid

Embodiment 5 obtains compound 152-160

Compound 152 - purchased from Transworld Chemical (South Melbourne, AUSTRALIA).

Compound 153 - purchased from Lancaster (Windham, NH).

Compound 154 - purchased from Avocado (Heysham, Lancashire, ENGLAND).

Compound 155 - purchased from Aldrich, Cat. No. 42919 (St. Louis, MO).

Compound 156 - purchased from Sigma-Aldrich (St. Louis, MO).

Compound 157 - purchased from Sigma (St. Louis, MO).

Compound 158 - purchased from Matrix Scientific (Columbia, SC).

Compound HPLC HPLC KF value Melting point range value CHNC CHNC CHNF CHNF

method

keep time

C C C C H

152 5.41 0

153 5.1 minutes 0.1%TF 0 69.76 5.46

A

154 6.2 minutes 0.1%TF 0 74.98 6.29

A

155 5.21 0

156 5.82 minutes 0.1%TF 0

A

157 5.42 0 184-186 73.67 5.3 72.56 4.91

158 5.47 0 110-112 74.36 5.82 74.39 5.66

159 5.56 minutes 63.47 5.39 62.65 5.13

1 60 5.30 0.3 67-70 73.45 5.32 73.08 5

Compound 160:

Potassium hydroxide (10.37 g, 184.8 mmol) was ground to a powder with a mortar and pestle and added to a 250 mL flask containing 75 mL of dimethyl sulfoxide and methyl 2-hydroxy-benzoate (7.03 g, 46.2 mmol). To this mixture was added benzyl bromide (7.91 g, 46.2 mmol) and allowed to mix with stirring for 4 hours. Water (100 mL) was added and the reaction was stirred for an additional 30 minutes. The reaction was then cooled to 0°C with an external ice bath and acidified to pH 1 with concentrated hydrochloric acid. The mixture was extracted with 3 x 230 mL ethyl acetate. The organic layers were combined and the solvent was removed under reduced pressure. The resulting yellow liquid was dissolved in ethyl acetate (50 mL) and washed with 2 x 30 mL of water, then 2 x 30 mL of brine. The organic layer was dried over sodium sulfate, filtered and the solvent was removed under reduced pressure. The resulting yellow liquid was dried under vacuum for several days to form a white crystalline solid. The solid product was collected and further dried under vacuum. The product was isolated (8.04 g, 76%) as a white crystalline solid, mp 67-70°C. Combustion analysis: measured value: C73.08, H5.37%; theoretical value of C ₁₄ H ₁₂ O ₃ is C73.45, H5.32%.

The preparation of embodiment 6 compound 160-167

Compound F was prepared according to a conventional procedure in which 2-hydroxybenzophenone was alkylated with a bromoalkyl ester in the presence of a base, followed by partial cleavage of the ester using potassium trimethylsiliconate.

(Compound 160) 6-(2-(2-hydroxybenzoyl)phenoxy)hexanoic acid:

A 250 mL round bottom flask equipped with a magnetic stir bar and reflux condenser was charged with 10.32 g (48.2 mmol) of 2,2'-dihydroxybenzophenone and 100 mL of dimethyl sulfoxide (DMSO). To the clear solution was added potassium hydroxide (2.91 g, 51.9 mmol) which had been ground to a powder. The reaction mixture was heated to 45°C until most of the solids had dissolved. The resulting red slurry was treated with 8.80 mL (11.04 g, 49.5 mmol) of ethyl 6-bromo-n-hexanoate. After stirring at 25°C for 20 hours, the clear reaction mixture was diluted with 1% aqueous hydrochloric acid and methyl tert-butyl ether (MTBE). The two layers were separated. The organic phase was washed with water (2 x 50 mL) and brine (1 x 40 mL), dried over sodium sulfate and concentrated. The residue was taken up in 100 mL of tetrahydrofuran (THF) and treated with potassium trimethylsiliconate (15.09 g, 118 mmol). The orange solution was stirred at 25°C for 20 hours, diluted to pH 7.5 with 4% aqueous hydrochloric acid and washed with MTBE. The organic phase was extracted with 3% aqueous sodium bicarbonate. The combined aqueous phases were acidified to pH 2 with 4% aqueous hydrochloric acid and extracted with 60 mL of MTBE. The organic phase was washed with brine (1 x 40 mL), dried over sodium sulfate and concentrated. The residue was purified by flash chromatography using 80% hexane/ethyl acetate (with the addition of 0.5% acetic acid). The product (4.2 g, 27%) was isolated as an off-white solid, mp 89-91°C. Combustion analysis _: measured value: C69.50, H6.04%; theoretical _value of _C19H20O5 : C: 69.50, H: 6.14%. ¹ H NMR (d6-DMSO): δ12.0, bs, 1H(COOH); δ11.5, bs, 1H(OH); δ7.5, t, 2H, (aryl H); δ7.4, dd , 1H (aryl H); δ7.3, dd, 1H (aryl H); δ7.15, d, 1H (aryl H); δ7.1, t, 1H (aryl H); δ7.0 , d, 1H (aryl H); δ6.9, t, 1H (aryl H); δ3.9, t, 2H, (CH ₂ at the Oα position); δ2.05, t, 2H (COOHα CH ₂ ); δ 1.4, m, 4H (the other two CH ₂ ); δ 1.0, p, 2H (CH ₂ in the middle of the chain).

The following compounds were prepared using the same method from appropriate starting materials: Compound 161, Compound 162, Compound 163, Compound 164, Compound 165, Compound 166 and Compound 167.

(Compound 161) Sodium 8-(2-(2-hydroxybenzoyl)phenoxy)octanoate: The title compound was prepared starting from 2,2'-dihydroxybenzophenone and ethyl 8-bromooctanoate, then prepared as follows It was converted to the sodium salt: the free acid (3.56 g, 9.99 mmol) was dissolved in 40 mL of isopropanol and treated with sodium hydroxide solution (1.7 mL) which was washed with sodium hydroxide (0.90 g, 22.5 mmol) and water (3.7 mL). Addition of isopropanol and MTBE caused precipitation of a solid. Heating the mixture caused most of the solids to dissolve. The remaining solid was removed by filtration. An off-white solid formed on cooling with dry ice was isolated by filtration and dried under reduced pressure. Combustion analysis: measured value: C65.02%, H6.22%; theoretical value of C ₂₁ H ₂₃ O ₅ Na is C66.00, H6.65%. ¹ HNMR (d6-DMSO): δ12.6, bs, 1H(OH); δ7.41, t, 1H, (aryl H); δ7.31, t, 1H (aryl H); δ7.27, dd, 1H (aryl H); δ7.15, dd, 1H (aryl H); δ7.03, d, 1H (aryl H); δ6.97, t, 1H (aryl H); δ6. 91, d, 1H (aryl H); δ6.65, t, 1H (aryl H); δ3.83, t, 2H, (CH ₂ at the Oα position); δ1.82, t, 2H (COONaα CH ₂ ); δ 1.3, m, 4H (the other two CH ₂ ); δ 1.0, m, 6H (CH ₂ in the middle of the chain). ¹³ C NMR(d6-DMSO)：198.59，177.35，161.35，156.10，134.56，131.98，131.78，129.55，128.57，123.57，120.18，118.00，117.09，112.51，67.74，37.87，28.83，28.35，28.27，25.84，24.87 .

(Compound 162) 5-(2-(2-Hydroxybenzoyl)-4-methoxyphenoxy)pentanoic acid (disclosed major isomer data): LC-MS analysis: confirmed m+1 Peak (345). ¹ H NMR analysis: (d6-DMSO): δ12.4, bs, 1H (COOH); δ11.9, bs, 1H (OH); δ7.47, t, 1H, (aryl H); δ7.26 , dd, 1H (aryl H); δ7.14, d, 1H (aryl H); δ7.13, d, 1H (aryl H); δ7.03, t, 1H (aryl H); δ6 .49, d, 1H (aryl H); δ6.42, dd, 1H (aryl H); δ3.95, t, 2H, (CH ₂ at the Oα position); δ3.79, s, 3H, ( CH ₃ O); δ 2.07, t, 2H (CH ₂ at the alpha position of COOH); δ 1.48, p, 2H (CH ₂ in the chain); δ 1.34, p, 2H (CH ₂ in the chain). ¹³ C NMR (d6-DMSO): 199.60, 174.18, 165.97, 163.34, 155.14, 135.14, 131.77, 128.29, 127.83, 120.46, 114.06, 112.69, 107.41, 100.70, 67.51, 55.76, 208.23

(Compound 163) 5-(2-(2-Hydroxybenzoyl)phenoxy)pentanoic acid: LC-MS analysis: Confirmation of m+1 peak (315). ¹ H NMR analysis: (d6-DMSO): δ11.9, bs, 1H (COOH); δ11.5, bs, 1H (OH); δ7.50, dt, 1H, (aryl H); δ7.48 , dt, 1H, (aryl H); δ7.35, dd, 1H (aryl H); δ7.25, dd, 1H (aryl H); δ7.14, d, 1H (aryl H); δ7.06, t, 1H (aryl H); δ6.96, d, 1H (aryl H); δ6.85, t, 1H (aryl H); δ3.93, t, 2H, (Oα δ2.06, t, 2H (CH ₂ at the alpha position of COOH); δ1.42, p, 2H (CH ₂ in _the chain); δ1.29, p, 2H (CH ₂ in the chain). ¹³ C NMR (d6-DMSO): 200.59, 174.15, 160.43, 155.71, 135.94, 132.69, 132.22, 128.58, 128.02, 121.50, 120.46, 119.06, 117.30, 112.67, 67.50, 33.05, 20.27

(Compound 164) 5-(2-(2-Hydroxy-5-methoxybenzoyl)-4-methoxyphenoxy)pentanoic acid: LC-MS analysis: m+1 peak was confirmed (375). ¹ H NMR analysis: (d6-DMSO): δ12.4, bs, 1H (COOH); δ12.0, bs, 1H (OH); δ7.25, d, 1H, (aryl H); δ7.21 , d, 1H, (aryl H); δ6.66, d, 1H (aryl H); δ6.62, dd, 1H (aryl H); δ6.48, d, 1H (aryl H); δ6.42, dd, 1H (aryl H); δ3.96, t, 2H, (CH ₂ at Oα position); δ3.81, s, 3H, (CH ₃ O); δ3.80, s, 3H , (CH ₃ O); δ2.08, t, 2H (CH ₂ at the alpha position of COOH); δ1.48, p, 2H (CH ₂ in the chain); δ1.34, p, 2H (CH ₂ in the chain ). ¹³ C NMR (d6-DMSO): 198.85, 174.20, 165.62, 164.14, 162.54, 157.11, 135.18, 130.22, 120.60, 114.44, 107.04, 105.51, 100.63, 99.24, 67.55, 27, 54.69, 33.7.7

(Compound 166) 4-(2-(2-Hydroxybenzoyl)phenoxy)butanoic acid: LC-MS analysis: m+1 peak was confirmed (333). ¹ H NMR analysis: (d6-DMSO): δ12.0, bs, 1H (COOH); δ7.46, m, 2H (aryl H); δ7.33, dt, 1H (aryl H); δ7. 29, d, 1H (aryl H); δ6.82, t, 1H (aryl H); δ3.77, t, 2H, (CH ₂ at Oα position); δ1.85, t, 2H (COOHα position CH ₂ ); δ 1.35, p, 2H (CH ₂ in the middle of the chain). ¹³ C NMR (d6-DMSO): 200.47, 173.92, 160.40, 155.57, 135.97, 132.64, 132.27, 128.64, 128.00, 121.52, 120.56, 119.10, 117.34, 112.62, 66.99, 29.525, 23.

(Compound 167) 4-(2-Chlorobenzoyl-4-methylphenoxy)butanoic acid: LC-MS analysis: m+1 peak was confirmed (333). ¹ H NMR analysis: (d6-DMSO): δ12.4, bs, 1H (COOH); δ12.0, bs, 1H (OH); δ7.23, d, 1H (aryl H, ortho to O) ; δ3.74, t, 2H, (CH ₂ at Oα position); δ2.25, s, 3H, (CH ₃ ); δ1.84, t, 2H (CH ₂ at COOH α position); δ1.33, p , 2H ( _CH2 in the middle of the chain). ¹³ C NMR (d6-DMSO): 198.76, 173.97, 165.63, 164.10, 162.58, 156.99, 135.11, 130.29, 120.55, 114.45, 107.14, 105.67, 100.67, 99.16, 67.03, 55.609, 235.5

The preparation of embodiment 7 compound 168-173

(Compound 168) 4-(2-Benzoyl-5-methoxyphenoxy)butanoic acid: Into a 100 mL mini-block tube equipped with a magnetic stir bar was charged 4.56 g (20.0 mmol) of 2-hydroxy- 4-Methoxybenzophenone, 2.70 mL (3.68 g, 18.9 mmol) ethyl 4-bromobutyrate and 40 mL dimethylformamide (DMF). Potassium carbonate (2.96 g, 21.4 mmol) was added to the clear solution. The reaction mixture was heated to 80 °C. After stirring at 25°C for 20 hours, the clear reaction mixture was diluted with water. The resulting solid was isolated by filtration. The solid was dissolved in 30 mL tetrahydrofuran (THF) and treated with 3.10 g (24.0 mmol) potassium trimethylsiliconate. The orange solution was stirred at 25°C for 20 hours, diluted to pH 7.5 with 4% aqueous hydrochloric acid and washed with MTBE. The organic phase was extracted with 3% aqueous sodium bicarbonate. The combined aqueous phases were acidified to pH 2 with 4% hydrochloric acid and extracted with 60 mL MTBE. The organic phase was washed with brine (1 x 40 mL), dried over sodium sulfate and concentrated. The resulting solid was purified by trituration using MTBE/hexanes. More product was separated from the mother liquor. LC-MS analysis: Confirmation of the m+1 peak (315). ¹ H NMR analysis: (d6-DMSO): δ12.0, bs, 1H (COOH); δ7.6, d, 2H, (phenyl H, ortho to CO); δ7.56, t, 1H (benzene group H, para-position of CO); δ7.44, t, 2H (phenyl H, meta-position of CO); δ7.35, d, 1H (aryl H, ortho-position of CO); δ6.64, m , 2H (aryl H, meta position of CO); δ3.88, t, 2H, (CH ₂ at Oα position); δ3.82, s, 3H, (CH ₃ O); δ1.84, t, 2H (CH ₂ at the alpha position of COOH); δ 1.53, p, 2H (CH ₂ in the middle of the chain). ¹³ C NMR (d6-DMSO): 195.08, 173.91, 163.17, 158.33, 138.84, 132.37, 131.37, 128.67, 128.24, 120.87, 105.87, 99.02, 66.89, 55.53, 29.45, 23.79.

Other delivery agents were prepared in the same way: Compound 169, Compound 170, Compound 171, Compound 172 and Compound 173.

(Compound 169) 4-(2-Benzoyl-4-chlorophenoxy)butyric acid: LC-M analysis: Confirmed m+1 peak (319). ¹ H NMR analysis: (d6-DMSO): δ11.9, bs, 1H (COOH); δ7.δ4, d, 2H, (phenyl H, ortho to CO); δ7.59, t, 1H (benzene δ7.51, dd, 1H (aryl H, para to CO); δ7.45, t, 2H (phenyl H, meta to CO); δ7.3δ, d , 1H (aryl H, ortho to CO); δ7.14, d, 1H (aryl H, meta to CO); δ3.87, t, 2H, (CH ₂ at Oα position); δ1.84 , t, 2H (CH ₂ at the alpha position of COOH); δ1.53, p, 2H (CH ₂ in the middle of the chain). ¹³ CNMR (dδ-DMSO): 194.37, 173.82, 154.74, 13δ.9δ, 133.42, 131.5δ, 130.05, 128.97, 128.δ2, 128.29, 124.48, 114.61, 67.38, 29.37, 23.79.

(Compound 170) 4-(2-Benzoyl-4-bromophenoxy)butanoic acid: LC-M analysis: Confirmed m+1 peak (363). ¹ H NMR analysis: (d6-DMSO): δ11.9, bs, 1H (COOH); δ7.6, m, 3H, (aryl H); δ7.60, t, 1H (phenyl H, CO para); δ7.49, dd, 1H (aryl H, ortho to CO); δ7.46, t, 2H (phenyl H, meta to CO); δ7.09, d, 1H (aryl H, the meta position of CO); δ3.89, t, 2H, (CH ₂ at the Oα position); δ1.82, t, 2H (CH ₂ at the COOHα position); δ1.53, p, 2H (the middle of the chain _CH2 ). ¹³ C NMR (d6-DMSO): 194.28, 173.81, 155.19, 136.97134.48, 133.42, 131.06, 130.48, 128.97, 128.62, 115.08, 112.02, 67.33, 29.35, 23.77.

(Compound 171) 4-(2-(2-chlorobenzoyl-5-methylphenoxy)butanoic acid: LC-M analysis: m+1 peak (333) was confirmed. ¹ H NMR analysis: (d6- DMSO): δ12.0, bs, 1H (COOH); δ7.54, d, 1H (aryl H); δ7.4, m, 2H (aryl H); δ7.33, dt, 1H (aryl H); δ7.29, d, 1H (aryl H); δ6.86, m, 2H (aryl H, ortho to O); δ3.77, t, 2H, (CH ₂ at Oα position); δ2.31, s, 3H, (CH ₃ ); δ1.85, t, 2H (CH ₂ at the alpha position of COOH); δ1.35, p, 2H (CH ₂ in the middle of the chain). ¹³ C NMR (d6-DMSO ): 193.31, 173.81, 158.34, 145.98, 141.38, 130.99, 130.56, 129.48, 129.43, 128.38, 127.00, 123.95, 121.46, 113.43, 66.95, 29.65, 23.70, 21.48.

(Compound 172) 4-(2-(2-chlorobenzoyl-4-methylphenoxy)butanoic acid: LC-M analysis: m+1 peak (333) was confirmed. ¹ H NMR analysis: (d6- DMSO): δ11.95, bs, 1H (COOH); δ7.43, m, 3H, (aryl H); δ7.34, m, 3H (aryl H); δ6.92, d, 1H (aryl δ3.74, t, 2H, (CH ₂ at Oα position); δ2.25, s, 3H, (CH ₃ ); δ1.84, t, 2H (CH at COOHα position ₂ ); δ1.33, p, 2H (CH ₂ in the middle of the chain). ¹³ C NMR (d6-DMSO): 193.92, 173.81, 156.15, 140.95, 135.37, 131.24, 130.40, 129.65, 129.56, 129.49, 128.62, 127.02 , 126.45, 112.95, 67.07, 29.65, 23.75, 19.80.

(Compound 173) 4-(2-Benzoyl-4-chloro-5-methylphenoxy)butanoic acid: LC-M analysis: Confirmed m+1 peak (333). ¹ H NMR analysis: (d6-DMSO): δ11.9, bs, 1H (COOH); δ7.δ1, d, 2H, (phenyl H, ortho to CO); δ7.57, t, 1H (benzene group H, para-position of CO); δ7.44, t, 2H (phenyl H, meta-position of CO); δ7.33, s, 1H (aryl H, ortho-position of O); δ7.14, s , 1H (aryl H, meta-position of O); δ3.87, t, 2H, (CH ₂ at Oα position); δ2.33, s, 3H, (CH ₃ ); δ1.81, t, 2H ( CH ₂ at the alpha position of COOH); δ 1.49, p, 2H (CH ₂ in the middle of the chain). ¹³ C NMR (dδ-DMSO): 194.31, 173.83, 154.78, 139.80, 137.39, 133.17, 128.91, 128.84, 128.51, 127.55, 124.69, 115.57, 67.32, 29.37, 23.80, 20.03.

Selective preparation of compound F

Compound F can be prepared selectively by Friedel-Crafts acylation of aromatic compounds:

Take an appropriate substituted phenol, and mix it with an appropriate bromoester, use _K2CO3 as _a base, and react the product with an appropriate aromatic acid chloride in the presence of _AlCl3 ; or take an appropriate substituted salicylic acid, and This is reacted _with the appropriate bromoester using _K2CO3 as the base. The product is converted to the acid chloride _SOCl2 , which is then reacted with the appropriate substituted benzene in the presence of _AlCl3 .

The preparation of embodiment 8 compound 174

Compound 174 was prepared in three steps:

A. O-acetyl-5-chlorosalicylic acid

10 g (57.9 mmol) of 5-chlorosalicylic acid was weighed in a 100 mL round bottom flask followed by the addition of acetic anhydride (12.8 mL, 115.9 mmol). The mixture was stirred for 5 minutes, then concentrated sulfuric acid (2 drops) was added. The reaction was refluxed for 3 hours. The progress of the reaction was monitored by HPLC. The reaction mixture was cooled to room temperature and poured into a beaker containing 2 N HCl (200 mL) to precipitate the product. The product was collected by vacuum filtration. A purity check by HPLC indicated the presence of impurities. The precipitate was stirred overnight in a 200 mL round bottom flask with water (150 mL). The insoluble solid was collected by vacuum filtration. Impurity check by HPLC showed no impurities in the crude product. The product was dried overnight in vacuo to obtain 12 g of O-acetyl-5-chlorosalicylic acid (56 mmol, 97% yield).

B. O-acetyl-5-chlorosalicyloyl chloride

Thionyl chloride (~100 mL) was charged to a 250 mL round bottom flask and stirred in an ice bath for 15 minutes. O-acetyl-5-chlorosalicylic acid (6.0 g, 27.9 mmol) was slowly added to the cooled thionyl chloride. DMF (2 drops) was added to the reaction mixture to aid dissolution of the acid. The reaction was stirred overnight to obtain a homogeneous solution. Excess thionyl chloride was distilled off in vacuo. The remaining residue was dried overnight in vacuo.

C. 3-(N-2-Hydroxy-5-chlorobenzoyl)alanine

β-Alanine (2.5 g, 28.0 mmol) was weighed in a 250 mL round bottom flask. Dichloromethane (100 mL) was added to the flask and the mixture was stirred for 5 minutes. Chlorotrimethylsilane (δ.06 g. 55.7 mmol) was added dropwise to the flask. The reaction was heated to reflux for 1.5 hours. The mixture was allowed to cool to room temperature and placed in an ice bath for 15 minutes. Triethylamine (8.5 g, 84.0 mmol) was slowly added to the cooled flask. O-acetyl-5-chlorosalicyloyl chloride (7.6 g, 27.9 mmol) was dissolved in dichloromethane (30 mL) and added to the reaction over 0.5 h. The reaction was stirred overnight and allowed to warm to room temperature. The course of the reaction was confirmed by HPLC. The solvent was evaporated under vacuum. The remaining residue was stirred in 2N NaOH (-100 mL) for 2 h and slowly heated to 60 °C. The solution was cooled to room temperature, then filtered by gravity filtration. The filtrate was slowly acidified with concentrated HCl until a precipitate formed. The crude product was collected when the mixture was at pH 6. The product was recrystallized using MeOH- _H2O . A purity check by HPLC indicated the presence of impurities. The product is subjected to multiple purification and recrystallization steps until the pure compound is obtained. The final product was stirred overnight in dichloromethane, collected by filtration and dried under vacuum overnight to obtain a pale pink powder (3.98 g, 16.3 mmol, 58.5% yield); mp 181-183 °C; ¹ H NMR (DMSO- d6) δ2.47-2.58(t, 2H), 3.44-3.54(q, 2H), 6.93-6.98(d, 1H), 7.39-7.44(dd, 1H), 7.91-7.96(d, 1H), 8.93 -9.01(t, 1H), 12.1-12.3(s, 1H). KF value = 1.615%. Anal. _Calcd . for _C10H10NO4Cl * _0.2220H2O : C, 48.50, H, 4.25, _N , 5.66. Found: C, 48.20; H, 4.03; N, 5.43.

Preparation of Example 9 Compound 175-178

Compound 175-4-(2-Benzyloxy-phenoxy)-butanoic acid:

Under an inert atmosphere, add 2-benzyloxy-phenol (8.0 g, 40 mmol), ethyl 4-bromobutyrate (5.7 mL, 40 mmol), potassium carbonate to a 250 mL flask equipped with a reflux condenser and a magnetic stirrer (7.2 g, 52 mmol) and ethanol (100 mL). The reaction mixture was stirred and heated to reflux for 8 hours. The reaction was cooled to room temperature and insoluble by-products were removed by suction filtration. To the filtrate was added 2 N aqueous sodium hydroxide solution (30 mL). The solution was heated to 50°C for 2 hours. The solution was cooled to room temperature, ethanol was removed under reduced pressure and the resulting solution was adjusted to pH9. The aqueous solution was washed with ethyl acetate (2 x 30 mL) and the residual ethyl acetate was removed under reduced pressure. The solution was cooled to 0°C with an external ice bath, then acidified to pH 2 with 6N aqueous hydrochloric acid. The precipitated product was collected by suction filtration and dried under vacuum. The product (7.2 g, 63%) was isolated as a white powder. ¹ H-NMR (400MHz, DMSO-d6): δ12.0, s, 1H (COOH); δ7.4, multiplet, 5H (benzylaryl H); δ7.0, multiplet, 2H (aryl H); δ6.9, multiplet, 2H (aryl H); δ5.0, s, 2H (benzyl CH ₂ ); δ4.0, t, 2H (CH ₂ at the alpha position of the phenoxy group); δ2 .4, t, 2H (CH ₂ at the alpha position of COOH); δ 1.9, multiplet, 2H (the rest of CH ₂ ).

Compound 1 76-(4-Benzyloxy-phenoxy)-acetic acid was purchased from Lancaster.

Compound 177-11-(2-benzyloxyphenoxy)undecanoic acid:

To a 250 mL Erlenmeyer flask was added freshly ground potassium hydroxide (4.2 g, 74.91 mmol) and 100 mL dimethylsulfoxide. 2-Benzyloxy-phenol (5 g, 24.97 mmol) and methyl 11-bromoundecanoate (7 g, 25.07 mmol) were added and the mixture was allowed to stir at room temperature overnight. Water (75 mL) was added and the solution was heated to 85°C and stirred for 3 hours. The reaction was cooled to room temperature and acidified to pH 2 with concentrated hydrochloric acid. The acidified solution was cooled to 4°C for 2 hours, then the precipitate was collected by suction filtration. The product was recrystallized from ethanol/water. The product (8.88 g, 93%) was isolated as a light brown solid, mp 62-63°C. Combustion analysis: measured value: C 74.71 H 8.08%; C ₂₄ H ₃₂ O ₄ theoretical value C 74.97, H 8.39%.

Compound 178-5-(4-benzyloxy-phenoxy)-pentanoic acid:

Under inert gas, add 4-benzyloxy-phenol (30.64g, 150mmol), ethyl 5-bromovalerate (31.99g, 150mmol), potassium carbonate (22.80g , 165mmol) and 270mL 2-butanone. The reaction mixture was heated to reflux for 23 hours, cooled, then diluted with ethyl acetate (150 mL) and extracted from water (500 mL). The organic layer was washed with water (1 x 250 mL) and brine (1 x 250 mL) and the solvent was removed under reduced pressure. The resulting oil was dried under vacuum for 4 days, during which time white crystals formed. The white crystals were dissolved in ethyl acetate (100 mL), washed with 1 N aqueous sodium hydroxide solution (3 x 50 mL), and the solvent was removed under reduced pressure. The resulting oil was dried under vacuum overnight to yield white crystals. The product was recrystallized from 1:1 ethanol:water and collected by suction filtration and dried under vacuum. The product was used without further purification.

To a 1 L round bottom flask equipped with a reflux condenser was added 5-(4-benzyloxy-phenoxy)-pentanoic acid ethyl ester (15.13, 46 mmol) and 2N aqueous sodium hydroxide solution (47 mL). The mixture was allowed to stir for 30 minutes. Water (200 mL) was added. The mixture was stirred for 20 minutes then heated to reflux for 2 hours to form a brown solution. The solution was rapidly cooled to room temperature by adding ice. The cooled solution was acidified with 2N aqueous hydrochloric acid (50 mL) and the resulting white precipitate was collected by suction filtration, washed with water (2 x 100 mL), hexanes (2 x 100 mL), and dried under vacuum overnight. The powder was ground and washed with hexane (1 x 250 mL) and diethyl ether (1 x 250 mL) to give a white powder. The powder was dissolved in a mixed solution of ethyl acetate (300 mL) and diethyl ether (200 mL) in a 1 L beaker. The solution was heated for 10 minutes, methanol (5 mL) was added, heated for an additional 10 minutes, then filtered through a plug of celite to give a clear yellow solution. The product was crystallized by slow addition of hexane. The crystals harvested first were collected by filtration and hexane (200 mL) was added to the mother liquor. The solution was then concentrated under reduced pressure to a volume of 400 mL and allowed to stand. A second crop of crystals was collected by filtration and combined with the first crop. The product (8.92g, 65%) was isolated as white crystals, mp 127-128°C. Combustion analysis: measured value C71.01H 6.98%; C ₁₈ H ₂₀ O ₄ theoretical value C71.98, H6.71%;

Compound No. HPLC retention time KF value Melting point range value CHNCC CHNCH CHNFC CHNFH 175 5.93 minutes 176 5.54 0 69.76 5.46 176 5.54 0 69.76 5.46 177 9.19 0 63-63 74.97 8.39 74.71 8.08 178 6.12 0 127-128 71.98 6.71 71.01 6.98

Example 10 Solid Oral Delivery of PYY[3-36] in Rats

A stock solution of PYY [3-36] (80 mg/mL) prepared in deionized water (PYY was purchased from Bachem California Inc. of Torrance, CA) was used.

Add about 0.08 mg/tablet (about 0.3 mg/kg) of PYY (about 1 μL) and mix with about 13.5 or 27 mg/tablet (50 or 100 mg/kg) of the free acid or sodium salt of the delivery agent compound as described below . The punches, punches, and dies of a Carver 4350 manual tablet press with a caplet model sold by Natoli Engineering Company, Inc. (St. Charles, Missouri) were treated with magnesium stearate (0.1%). About 13.58 or about 27.08 mg of the blended powder was filled into a die and made into microbeaded tablets under a pressure of about 1000 psi bar. The resulting solid dosage form was about the size of a standard size 9 capsule (about 2.65mm in diameter and about 8.40mm in length) for the 27.08mg size, and about 2.65mm in diameter and about 4.20mm in length for the 13.58mg solid.

Male Sprague Dawley rats (about 260 to about 280 g) were fasted overnight and then anesthetized by standard _CO inhalation techniques for about 10 to 30 seconds to produce anesthesia in about less than 1 minute, preferably about 10 to about 30 seconds state.

Apply the oral administration tube. The dosing tube is inserted into the rat's mouth and carefully passed through the rat's pharynx and esophagus about 8 cm to about 15 cm, depending on the weight of the rat (typically about 11 cm). The solid dosage form is released into the end of the esophagus and/or stomach by depressing the plunger of the oral administration tube.

Blood samples were typically collected retro-orbitally at time = 0, 15, 30, 60 and 90 minutes. Serum PYY concentrations were quantified using the PYY[3-36] radioimmunoassay (Catalog #RK-059-02, Phoenix Pharmaceuticals, Inc., Belmont, CA). The mean of the results for each group of animals was calculated for each time point. The maximum of these means (ie mean peak serum PYY[3-36] concentration ± standard deviation (SD)) is listed below.

Table 1. Oral Administration of PYY[3-36] in Rats

delivery agent compound Application method Compound dose (mg/kg) PYY(3-36) dose (mg/kg) Mean serum peak value (pg/mL) ± SD of PYY 23-sodium salt Oral, solid dose, 1 tablet per animal 100 0.3 427.4±258.7 121-sodium salt Oral, solid dose, 1 tablet per animal 100 0.3 897.1±257.3 121-free acid Oral, solid dose, 1 tablet per animal 50 0.3 161.7±148.5

174-sodium salt Oral, solid dose, 1 tablet per animal 100 0.3 675±427.1 174-free acid Oral, solid dose, 1 tablet per animal 100 0.3 0

Example 11 Liquid Oral Delivery of PYY[3-36] in Rats

Oral gavage (PO) dosing solutions of transmitter compounds and peptide YY residues 3-36 (PYY[3-36]) (purchased from Bachem California Inc. of Torrance, CA) were prepared in deionized water as follows.

A stock solution of PYY[3-36] (80 mg/mL) was prepared in deionized water. Compositions for oral administration comprising 200 mg/kg of delivery agent compound and 0.3 mg/kg of PYY were prepared in aqueous solution. A solution of compound 23 was prepared with one equivalent of sodium hydroxide to convert the free acid transfer agent to its sodium salt.

Male Sprague-Dawley rats weighing 240-320 g were fasted for up to 24 hours prior to testing and administered ketamine (44 mg/kg) and sorazine (1.5 mg/kg) by intramuscular injection prior to administration of the test item. The anesthetized animals were then administered the test item by oral gavage. A drug solution was administered to a dosing group of 5 animals. For oral feeding (PO), an 11 cm Rusch 8 French catheter was attached to a 1 mL syringe with a micropipette tip. The syringe is filled with the dosing solution by drawing the solution through the catheter and then wiping the catheter dry. A catheter is placed in the esophagus, leaving 1 cm of tubing outside the incisors. The dosing solution is administered by depressing the plunger of the syringe.

Blood samples were collected serially from the tail artery or by cardiac puncture, typically at time = 0, 15, 30, 45, 60 and 90 minutes. Serum PYY concentrations were quantified using the PYY[3-36] radioimmunoassay (Catalog #RK-059-02, Phoenix Pharmaceuticals, Inc., Belmont, CA). The mean of the results for each group of animals was calculated for each time point. The maximum of these means (ie mean peak serum PYY[3-36] concentrations ± standard deviation (SD)) are listed in Table 2 below. No significant PYY[3-36] was detected in blood when animals were orally administered only PYY[3-36].

Table 2. Oral Administration of PYY[3-36] in Rats (Liquid)

compound Application method Compound dose (mg/kg) PYY(3-36) dose (mg/kg) Mean serum peak value (pg/mL) ± SD of PYY twenty three Oral (liquid dose) 200 0.3 788.198±50.59 151 Oral (liquid dose) 200 0.3 801.96±290.61

158 Oral (liquid dose) 200 0.3 1065±75.352 160 Oral (liquid dose) 200 0.3 370.39±306.29 160 Oral (liquid dose) 200 0.3 631.96±316.16 160 Oral (liquid dose) 200 0.3 705.106±75.906 161 Oral (liquid dose) 200 0.3 340.95±228.946 174 Oral (liquid dose) 200 0.3 1262.26±313.58

Example 12 Oral Delivery of Human Recombinant Insulin in Rats

Insulin (human recombinant) was purchased from ICN Biomedicals (Aurora, OH) as a bulk powder. To prepare stock solutions, insulin was dissolved in deionized water (pH~6.5) to obtain a concentration of 15 mg/mL. Stock solutions were kept frozen at -20°C in 1.0 mL aliquots until use. For dosing solutions, the delivery agent compounds were dissolved in deionized water to obtain a final concentration of 200 mg/mL. The free acid form of the delivery agent was converted to the sodium salt by adding one equivalent of sodium hydroxide. The solution was vortexed, sonicated, and heated, and if necessary, additional μL amounts of sodium hydroxide were added to achieve consistent solubility. The solution was adjusted to pH 3.5-8.5 by adding hydrochloric acid or sodium hydroxide. Insulin stock solution (typically 66.7 μL) was then added to the delivery agent solution to obtain a final concentration of 0.5 mg/mL. After dissolving and adding the drug, the solution was adjusted to the final volume by adding deionized water.

Insulin alone or in combination with Emisphere delivery agent was administered to male Sprague-Dawley rats by oral gavage (PO). Typically, rats are fasted for 18-24 hours prior to dosing. For dosing, a Rusch 8 French catheter was cut to a length of 11 cm and fit into a 1 mL syringe. Fill the syringe with dosing solution and wipe the catheter dry of excess solution. A catheter was inserted into the rat's mouth and into the esophagus (10.0 cm). The rat was held in a vertical position and the drug solution was released by depressing the plunger of the syringe.

Sample Collection and Processing: Insulin

管(TerumoCorporation，Tokyo，Japan)中。允许样品在4℃凝固～20分钟。凝固后，样品在6℃、10,000rpm下离心4分钟以便分离血清。将血清收集于eppendorf管中并且在-20℃下冷冻直到分析。During blood sampling, rats were briefly (~10 sec) exposed to carbon dioxide immediately before each sampling time point until collapsed. For blood sampling, a 77-mm capillary was inserted into the retro-orbital fossa. Typically, blood samples were collected pre-dose (time 0) and 15, 30, 45 and 60 minutes post-dose. Samples were collected in a coagulant containing coagulant (red cap, serum separator tube)

tube (Terumo Corporation, Tokyo, Japan). Samples were allowed to freeze at 4°C for ~20 minutes. After coagulation, the samples were centrifuged at 10,000 rpm for 4 minutes at 6°C to separate the serum. Sera were collected in eppendorf tubes and frozen at -20°C until analysis.

Sample Collection and Processing: Whole Blood Glucose

To determine pharmacodynamic response, a handheld blood glucose meter (OneTouch Ultra, (Johnson & Johnson, New Brunswick, New Jersey)) to measure blood glucose in whole blood. Samples were collected from the retro-orbital fossa (see Sample Collection and Handling: Insulin) or the tail artery (ie, tail snip). For tail snipping, the tip of the tail was severed approximately 5 mm from the tip using a scalpel blade. After the first drop of blood is discarded, a small sample (~5-10 μL) is touched to a glucometer test strip (OneTouch Ultra, LifeScan) and blood glucose readings are usually taken by the instrument. For each subsequent sampling time point, clots formed at the tail tip were disrupted and fresh samples collected. Typically, samples were collected pre-dose (time 0) and 15, 30, 45 and 60 minutes post-dose.

Table 3. Oral (liquid dose) administration of insulin to rats

compound Application method Delivery agent compound dose (mg/kg) Insulin dose (mg/kg) Relative to the maximum percentage of control ± SD decline twenty four Oral (liquid dose) 200 0.5 -14.73±17.64 25 Oral (liquid dose) 200 0.5 -14.81±12.99 26 Oral (liquid dose) 200 0.5 -25.93±14.86 27 Oral (liquid dose) 200 0.5 -25.40±30.61 28 Oral (liquid dose) 200 0.5 -11.41±18.92 29 Oral (liquid dose) 200 0.5 -29.25±6.97 140 Oral (liquid dose) 100 1 Cmax＝81.16±114.98μIU/mL 141 Oral (liquid dose) 100 1 Cmax＝204.05±60.88μIU/mL 142 Oral (liquid dose) 100 1 Cmax＝118.16±72.75μIU/mL 145 Intracolon (liquid dose) 50 0.1 Cmax＝15.03±7.80μIU/mL 145 Oral (liquid dose) 100 1 Cmax＝3.92±5.62μIU/mL 160 Oral (liquid dose) 200 0.5 Cmax＝74±7.5μIU/mL

165 Oral (liquid dose) 200 0.5 -33.0 166 Oral (liquid dose) 200 0.5 -5.7 167 Oral (liquid dose) 200 0.5 -21.2 167 Oral (liquid dose) 200 0.5 -17.7 167 Oral (liquid dose) 200 0.5 -26.2 167 Oral (liquid dose) 200 0.5 -17.8 167 Oral (liquid dose) 200 0.5 -22.7

Example 13 Human Zinc Insulin - Oral Delivery

Oral administration (PO) compositions of the delivery agent compound and human zinc insulin (minimum 26 IU/mg, purchased from Calbiochem-Novabiochem Corp, La Jolla, CA) were prepared in deionized water. 500 mg of the delivery agent compound was added to 1.5 mL of water. The free acid of the transfer agent compound was converted to the sodium salt by stirring the resulting solution and adding one equivalent of sodium hydroxide. The solution was vortexed, then heated (approximately 37°C) and sonicated. Adjust the pH to about 7 to 8.5 with NaOH or HCl. Additional NaOH was added if necessary to achieve consistent solubility and the pH was readjusted to about 7 to 8.5. Water was then added to bring the total volume to approximately 2.4 mL and vortexed. About 1.25 mg from insulin stock solution (15 mg/mL, prepared from 0.5409 g insulin and 18 mL deionized water, adjusted to pH 8.15 with HCl and NaOH and obtained a clear solution with 40 mL concentrated hydrochloric acid, 25 mL 10N NaOH and 50 mL 1N NaOH) Insulin was added to the solution and mixed by inversion. Final delivery agent compound doses, insulin doses and dose volumes are listed.

Male Sprague-Dawley rats weighing approximately 200-250 g were fasted for 24 hours and administered ketamine (44 mg/kg) and chlorpromazine (1.5 mg/kg) 15 minutes before dosing and again requiring maintenance of anesthesia. A drug solution was administered to a dosing group of 5 animals. For oral administration, an 11 cm Rusch 8 French catheter was attached to a 1 mL syringe with a micropipette tip. The syringe is filled with the dosing solution by drawing the solution through the catheter and then wiping the catheter dry. A catheter is placed in the esophagus, leaving 1 cm of tubing outside the incisors. The dosing solution is administered by depressing the plunger of the syringe.

Blood samples were collected serially from the tail artery, typically at time = 15, 30, 60, 120 and 180 minutes. Serum insulin levels were determined using an insulin ELISA assay kit (Kit #DSL-10-1600, DiagnosticSystems Laboratories, Inc., Webster, TX), a standard method modified to optimize sensitivity and a standard curve for the sample volumes and concentrations used in this method the linear range. The serum human insulin concentration (μU/mL) of each of the five animals in each dosing group was determined at each time point. Five values for each time point were averaged and serum insulin concentrations were plotted against time. (Previous trials have shown that oral administration of human insulin alone has no measurable levels of human insulin). The maximum values (peak values) and areas under the curve (AUC) are listed in Table 4 below. For Blood Glucose the ONE TOUCH (Life Scan, Johnson & Johnson, New Brunswick, New Jersey) Percent change from baseline.

Table 4. Insulin - Oral Delivery

delivery agent compound Delivery agent compound dose (mg/kg) Insulin dose (mg/kg) Volume dose (ml/kg) C _max (glucose) (μU/mL) AUC Glucose (% control) 123 200 0.50 107.3 7440 125 200 0.50 98.3 7687.5 123 200 0.50 100.3 7447.5 115 200 0.50 83.3 3232.5 116 200 0.50 89.5 3292.5 118 200 0.50 90.5 4327.5 124 200 0.50 87.8 1582.5 134 200 0.50 81.5 3817.5 136 200 0.50 91.5 4507.5 138 200 0.50 93.4 6907.5 124 200 0.50 59.9 112.5 152 200 0.5 -29.3% 153 200 0.5 -7.1% 154 200 0.5 -7.9% 159 200 0.5 -6.6% 159 200 0.5 9.1 -36.5%

Example 14 Insulin-pulmonary delivery

A pharmaceutical composition of the delivery agent compound and human insulin is prepared in water. Typically, 1.5 mg of the transfer agent compound is added to deionized water to make a volume of 1.0 mL, and the solution is vortexed. The free acid was converted to the sodium salt using the sodium salt of the delivery agent or by stirring the resulting solution and adding one equivalent of sodium hydroxide (10N) and diluting with water. The solution was vortexed, then heated (approximately 37°C) and sonicated. The pH was adjusted to between about 7.0 and 8.5 with NaOH or HCl. 75 μL of human insulin stock solution (2 mg/mL) was added to the solution. (The stock solution was prepared as follows: 3 mL of a pH 3.0 HCl solution in deionized water was added to 0.02 g of insulin. The pH of the resulting solution was adjusted to below 3.0 (approximately 2.6) with HCl and NaOH until the solution was clear. and HCl to bring the pH to 7.6. Make a final volume of 10 mL with pH 7.5 deionized water. Final pH is 7.59). Water was then added to bring the total volume to 2.0 mL, and the solution was gently inverted several times. The solution can be used immediately in the method of administration, or the solution can be placed in a 37°C water bath for 1 hour prior to administration. Final delivery agent compound doses, insulin doses and volume doses are listed in Table 5 below.

Typical dosing and sampling methods are as follows. Male Sprague-Dawley rats weighing 200-250 g were fasted for 24 hours and administered ketamine (44 mg/kg) and Chlorpromazine (3.0 mg/kg). A drug solution was administered to a dosing group of 5 animals. A control group of 5 animals was administered insulin alone. A lighted rodent catheter dripper (purchased from Penn Century, Inc., Pittsburgh, PA) was filled with drug solution and inserted into the larynx until the needle entered the esophagus (identified visually). The dosing solution is administered by depressing the plunger.

Blood samples were collected serially from the tail artery, typically at 5, 15, 30, 60 and 120 minutes post-dose. Serum insulin levels were determined using an insulin ELISA assay kit (Kit #DSL-10-1600, DiagnosticSystems Laboratories, Inc., Webster, TX), a standard method modified to optimize sensitivity and a standard curve for the sample volumes and concentrations used in this method the linear range. The serum human insulin concentration (μU/mL) of each of the five animals in each dosing group was determined at each time point. Five values for each time point were averaged and serum insulin concentrations were plotted against time. The ratio of the area under the curve (AUC) for the test and control groups is listed below. The ratio of the maximum serum insulin concentration (C _max ) of the test and control groups is also listed below.

Table 5. Pulmonary Delivery of Insulin

Delivery agent

Delivery Agent Compound Dose Insulin Dose

    Volume Dose (mL/kg)

_Cmax

compound

(mg/kg) (mg/kg)

174 0.4 16 0.03 18.36±19.18

Example 15 Oral and Intracolonic Heparin Delivery

Oral gavage (PO) and/or intracolonic (IC) dosing solutions containing the delivery agent compound and heparin sodium USP are prepared in 25% propylene glycol in water. The sodium salt of the compound is used. Typically, the delivery agent compound and heparin (approximately 166-182 IU/mg) are mixed in dry powder form by vortexing. This dry mixture was dissolved in 25% v/v propylene glycol in water, vortexed and placed in a sonicator (approximately 37°C). The pH was adjusted to about 7 (6.5-8.5) with aqueous NaOH (2N). The dosing solution was sonicated to produce a clear solution. Adjust the final volume to 3.0 mL. The final delivery agent compound doses, heparin doses and volume doses are listed in Table 6 below.

Typical dosing and sampling methods are as follows. Male Sprague-Dawley rats weighing 275-350 g were fasted for 24 hours and anesthetized with ketamine (88 mg/kg) intramuscularly immediately prior to dosing. A drug solution was administered to a dosing group of 5 rats. For oral gavage (PO) administration, an 11 cm Rusch 8 French catheter was attached to a 1 mL syringe with a micropipette tip. The syringe is filled with the drug solution by drawing the solution through the catheter, and the catheter is then wiped dry. Place the catheter into the esophagus, leaving 1 cm of tubing outside the rat's incisors. The solution is administered by depressing the plunger of the syringe. For intracolonic (IC) administration, a 7.5 cm 8fr Rusch catheter was attached to a 1 mL syringe with a micropipette tip. The dosing catheter is inserted through the anus and into the colon until the tube is no longer visible. Slowly inject the dosing solution into the colon.

Citrated blood samples were typically collected by cardiac puncture at time = 0.25, 0.5, 1.0 and 1.5 hours after administration of ketamine (88 mg/kg). Heparin activity was determined by applying the activated partial thromboplastin time (APTT) according to the method of Henry, J.B., Clinical Diagnosis and Management by Laboratory Methods, Philadelphia, PA, W.B. Saunders (1979). Previous studies suggested a baseline value of around 20 seconds. The results were averaged for each time point of 5 rats per group. The maximum values are listed in Table 6 below.

Table 6. Oral/Intracolonic Delivery of Heparin

Delivery Agent Administration Volume Dose Compound Heparin Dose APTT Mean Peak (sec) ± SD pH

Compound Method (mL/kg) Dose (mg/kg) (mg/kg)

114 IC 1 50 25 42.90±8.70 7.61

140 IC 1 50 25 23.49±6.12 7.67

141 IC 1 50 25 52.40±21.54 7.62

143 IC 1 50 25 114.69±121.62 7.18

145 IC 1 50 25 134.42±99.03 6.93

151 PO 3 300 100 252.09±107.13

151 IC 1 50 25 2.36±1.27 (anti-factor Xa) 7.06

151 IC 1 25 25 3.39±3.07 (anti-factor Xa) 7.23

160 IC 1 50 25 131±154 (T _max = 90 minutes)

Example 16 Parathyroid Hormone Delivery (PTH 1-34) Oral/Intracolonic Delivery

Oral gavage (PO) and/or intracolonic (IC) administration solutions of delivery agent compounds and human parathyroid hormone residues 1-34 (PTH) are prepared in water. The sodium salt of the delivery agent compound is used. Typically, solutions of the compounds are prepared in water and stirred, one equivalent of sodium hydroxide (1.0 N) is added when preparing the sodium salt. Final drug solutions were prepared by mixing compound and PTH stock solutions (PTH was purchased from Eli Lilly and Co., Indianapolis, IN) (typically at a concentration of 5 mg PTH/mL) and diluted to the desired volume (typically 3.0 mL). Final compound doses, PTH doses and volume doses are listed in Table 7 below.

Typical dosing and sampling methods are as follows. Male Sprague-Dawley rats weighing 200-250 g were fasted for 24 hours and administered ketamine (44 mg/kg) and chlorpromazine (1.5 mg/kg) 15 minutes prior to dosing. A drug solution was administered to a dosing group of 5 rats. For oral gavage (PO) administration, an 11 cm Rusch 8 French catheter was attached to a 1 mL syringe with a micropipette tip. The syringe is filled with the drug solution by drawing the solution through the catheter, and the catheter is then wiped dry. Place the catheter into the esophagus, leaving 1 cm of tubing outside the rat's incisors. The solution is administered by depressing the plunger of the syringe. For intracolonic (IC) administration, 7.5 cm Rusch catheters (French 8 or 6) are suitable for syringes with Eppendorf micropipette tips. The syringe draws the solution through the catheter to fill it with the drug solution. Wipe the catheter dry. Apply K-Y gel to the tip, avoiding contact with the eye of the tube, and insert the tube through the anus into the colon until the tube is no longer visible. The solution is injected by depressing the plunger of the syringe and the tubing is removed.

Blood samples were collected serially from the tail artery, typically at time = 0, 15, 30, 45, 60 and 90 minutes for oral administration and typically at time = 0, 10, 20, 30, 60 and 90 minutes for IC administration minute. Serum PTH concentration was quantified by PTH radioimmunoassay kit (kit #RIK 6101 purchased from Peninsula Laboratories, Inc. San Carlos, CA). Previous studies have shown a baseline value of around 0. The results were averaged for each time point of 5 rats per group. The maximum values are listed in Table 7 below.

Table 7. Oral/intracolonic delivery of PTH in rats

delivery agent compound Application method Volume dose (mL/kg) Compound dose (mg/kg) PTH dose (μg/kg) Serum [PTH] mean peak ± SD pH 113 oral 1 100 200 780.77±439.92 8.18 113 oral 1 100 200 53.51±39.55 8.09 114 oral 1 100 200 135.78±136.97 8.41

Example 17 Interferon - oral delivery

Prepare delivery agent compounds and interferon alpha-1 (IFN) in deionized water ( Dosing solution available from InterMune, Inc. of Brisbane, CA). The free acid of the transfer agent compound was converted to the sodium salt with one equivalent of sodium hydroxide. Typically, a solution of the transfer agent compound is prepared in water and stirred, with one equivalent of sodium hydroxide (1.0 N) added when preparing the sodium salt. The mixture was vortexed and placed in a sonicator (approximately 37°C). The pH was adjusted to about 7.0 to 8.5 with aqueous NaOH. The mixture is vortexed to produce a homogeneous suspension or solution, sonication and heating are also applied if necessary. Additional NaOH was added if necessary to achieve uniform solubility and the pH was adjusted to about 7.0 to 8.5. The delivery agent compound solution was mixed with the IFN stock solution (approximately 22.0 to 27.5 mg/mL in phosphate buffered saline) and diluted to the desired volume (typically 3 mL). Final delivery agent compound and IFN doses and dose volumes are listed in Table 8 below.

Typical dosing and sampling methods are as follows. Male Sprague-Dawley rats weighing 200-250 g were fasted for 24 hours and administered ketamine (44 mg/kg) and chlorpromazine (1.5 mg/kg) 15 minutes before dosing and again requiring maintenance of anesthesia. A drug solution was administered to a dosing group of 5 rats. An 11 cm Rusch 8 French catheter was attached to a 1 mL syringe with a micropipette tip. The syringe is filled with the drug solution by drawing the solution through the catheter, and the catheter is then wiped dry. A catheter is placed in the esophagus, leaving 1 cm of tubing outside the incisors. The dosing solution is administered by depressing the plunger of the syringe.

Blood samples were collected serially from the tail artery, typically at time = 0, 15, 30, 45, 60 and 90 minutes. Serum IFN concentrations were quantified using the Cytoscreen Immunoassay Kit for Human IFN-α (Catalog #KHC4012 from Biosource International, Camarillo, CA). Previous studies have shown a baseline value of around 0. The results were averaged for each group of animals at each time point. The maximum of these means (ie, mean peak serum IFN concentration) is listed in Table 8 below.

Table 8. Interferons - Oral Delivery

delivery agent compound Delivery agent compound dose (mg/kg) IFN dose (mg/kg) Volume dose (mL/kg) Serum [IFN] mean peak (ng/mL) ± SD pH 141 200 1 1 0.73±0.44 8.29 147 200 1 1 1.27±0.60 8.45 174 200 1 1 0.5±0.57 174 200 1 1 0.18±0.17 174 200 1 1 3.96±2.72 174 200 1 1 17.4±9.12

Example 18 Oral Delivery of Salmon Calcitonin (sCT)

Orally administered compositions of the delivery agent compound and salmon calcitonin (sCT) were prepared in water. Typically, 450 mg of the delivery agent compound is added to 2.0 mL of water. The free acid was converted to the sodium salt using the sodium salt of the compound or by stirring the resulting solution and adding one equivalent of sodium hydroxide (1.0 N) and diluting with water. The solution was vortexed, then heated (approximately 37°C) and sonicated. The pH was adjusted to about 7 (6.5 to 8.5) with NaOH or HCl. A stock solution containing 90 μg sCT was added to the solution. Water was then added to bring the total volume to approximately 3 mL (varies depending on the solubility of the delivery agent compound). Final delivery agent compound doses, sCT doses and volume doses are listed in Table 9 below.

Male Sprague-Dawley rats weighing 200-250 g were fasted for 24 hours and administered ketamine (44 mg/kg) and chlorpromazine (1.5 mg/kg) 15 minutes prior to dosing. One dosing solution was administered to a dosing group of 5 rats. For oral administration, an 11 cm Rusch 8 French catheter was attached to a 1 mL syringe with a micropipette tip. The syringe is filled with the dosing solution by drawing the solution through the catheter and then wiping the catheter dry. Place the catheter into the esophagus, leaving 1 cm of tubing outside the rat's incisors. The solution is administered by depressing the plunger of the syringe.

Blood samples were collected serially from the tail artery, typically at time = 0, 10, 20, 30, 60 and 90 minutes. Serum sCT was determined by measuring with an EIA kit (kit #EIAS-6003 purchased from Peninsula Laboratories, Inc., San Carlos, CA), the standard method of which was modified as follows: Incubate with 50 μL of peptide antibody for 2 hours (in the dark shaking), plates were washed, serum and biotinylated peptides were added and diluted with 4 mL of buffer, and shaken overnight in the dark. Numbers were adjusted according to the baseline value obtained at time=0. The results were averaged for each group of animals at each time point. The maximum values are listed in Table 9 below.

Table 9 Oral delivery of salmon calcitonin (sCT)

delivery agent compound Volume dose (mL/kg) Compound dose (mg/kg) SCT dose (μg/kg) Serum sCT average peak (pg/mL) ± SD (SE) 174 150 30 1 182.83±184.82 174 150 30 1 198.21±205.15 174 150 30 1 70.81±118.47

Example 19 Oral/colonic delivery of recombinant human growth hormone (rhGH)

Oral gavage (PO) and/or intracolonic (IC) administration solutions of delivery agent compounds and rhGH (rhGH were purchased from Novartis, Basel, Switzerland) were prepared in phosphate buffered saline. The sodium salt of the delivery agent compound is obtained by reacting the free acid with one equivalent of sodium hydroxide. Final drug solutions were prepared by mixing compounds with rhGH stock solution (15 mg rhGH/mL) and diluting to desired volume (typically 3 mL). Compounds and doses of rhGH are listed in Table 10 below.

Typical dosing and sampling methods are as follows. Male Sprague-Dawley rats weighing 200-250 g were fasted for 24 hours and administered ketamine (44 mg/kg) and chlorpromazine (1.5 mg/kg) 15 minutes prior to dosing. A drug solution was administered to a dosing group of 5 rats. For oral gavage (PO) administration, an 11 cm Rusch 8 French catheter was attached to a 1 mL syringe with a micropipette tip. The syringe is filled with the drug solution by drawing the solution through the catheter, and the catheter is then wiped dry. Place the catheter into the esophagus, leaving 1 cm of tubing outside the rat's incisors. The solution is administered by depressing the plunger of the syringe. For intracolonic (IC) administration, 7.5 cm Rusch catheters (French 8 or 6) are suitable for syringes with Eppendorf micropipette tips. The syringe draws the solution through the catheter to fill it with the drug solution. Wipe the catheter dry. Apply K-Y jelly to the tip, avoiding contact with the eye of the tube, and insert the tube through the anus into the colon until the tube is no longer visible. The solution is injected by depressing the plunger of the syringe and the tubing is removed.

Blood samples were collected serially from the tail artery or retro-orbital pit, typically at time = 0, 15, 30, 45, 60 and 90 minutes for oral administration and at time = 0, 10, 20, 30, 60 and for IC administration 90 minutes. Samples were collected in a coagulant containing coagulant (red cap, serum separator tube) in tubes (Terumo Corporation, Tokyo, Japan). Samples were allowed to freeze at 4°C for ~20 minutes. Serum rhGH concentrations were quantified by rhGH Immunoassay Assay Kit (Kit #K1F4O15 purchased from Genzyme Corporation Inc., Cambridge, MA). Pool 5 samples for each time period. Previous studies have shown a baseline value of around 0.

The maximum concentration for each group is listed in Table 10 below.

Table 10. Oral/intracolonic delivery of rhGH in rats

Delivery agent compound administration volume dose compound rhGH mean peak serum rhGH

Drug Method (mL/kg) Dose (mg/kg) Dose (mg/kg) (ng/mL)

160 PO 1 200 3 3 -

161 PO 1 200 3 1.033(±2.31)(T _max ＝15 points

bell)

174 PO 1 200 3 57.42

All publications, references, patents, and published patent applications are incorporated herein by reference.

Claims (5)

1. A compound selected from the group consisting of: Compound 23 or its sodium salt, Compound 24

Compound 25

Compound 26 Compound 27

Compound 28 2. A composition comprising: (A) at least one bioactive agent; and (B) the compound of claim 1, Wherein the bioactive agent is PYY, PYY agonist or insulin. 3. The composition of claim 2, wherein said bioactive agent is PYY [3-36]. 4. The composition of claim 2, wherein said bioactive agent is recombinant insulin. 5. Use of the composition according to any one of claims 2-4 in the preparation of a medicament.