RU2530889C2 - Compounds and formulations for active substance delivery - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to compounds or their pharmaceutically acceptable salts which can be applicable for biologically active substances delivery.

EFFECT: invention refers to pharmaceutical formulations containing the above compounds, and to a method for administering the biologically active substances.

17 cl, 10 tbl, 19 ex

Description

[1] This application claims priority according to provisional patent application US No. 60 / 576,088, filed June 1, 2004, provisional application for US patent No. 60 / 576,397, filed June 1, 2004, provisional application for US patent No. 60 / 576,105, filed June 1, 2004, provisional application for US patent No. 60 / 571,090, filed May 14, 2004, provisional application for US patent No. 60 / 571,092, filed May 14, 2004, provisional application for US patent No. 60 / 571,195, filed May 14, 2004, provisional application for US patent No. 60 / 571,194, filed May 14, 2004, provisional application for US patent No. 60 / 571,093, filed May 14, 2004, provisional application for US patent No. 60 / 571,055, filed May 14, 2004, provisional application for US patent No. 60 / 571,151, filed May 14, 2004, provisional application for US patent No. 60 / 571,315, filed May 14, 2004, provisional application for US patent No. 60 / 571,144, filed May 14, 2004, and provisional patent application US 60 / 571,089, filed May 14, 2004, a description of each of which is fully incorporated into the description of this application by reference.

Technical field

[2] The present invention relates to compounds and compositions for the delivery of active substances, such as biologically or chemically active substances, to a target (organ). These compounds are well suited for the formation of non-covalent mixtures with active substances for oral or other route of administration to animals. Methods for preparing and administering such compositions are also described.

BACKGROUND OF THE INVENTION

[3] Conventional methods for the delivery of active substances are often severely limited by biological, chemical, and physical barriers. Typically, these barriers are imposed by the environment through which delivery occurs, the environment directly around the delivery target, and / or the target itself. Biologically and chemically active substances are especially vulnerable to such barriers.

[4] Upon the delivery of biologically and chemically active pharmacological and therapeutic agents into the animal’s body, the body creates barriers. Examples of physical barriers are skin, epithelium, lipid bilayers and membranes of various organs, which are relatively impervious to certain active substances, but which must be overcome in order to reach a delivery target, for example, the circulatory system. Chemical barriers include, but are not limited to, changes in pH in the gastrointestinal (GI) tract and destruction by enzymes.

[5] These barriers are of particular importance in the development of oral delivery systems. The oral delivery of many active substances would be chosen by the method of introducing them into the animal organism, if not for biological, chemical and physical barriers. Numerous substances that are traditionally not orally administered include biologically or chemically active peptides, for example, calcitonin or insulin; polysaccharides, such as mucopolysaccharides, including, without limitation, heparin; heparinoids; antibiotics and other organic substances. In the gastrointestinal tract under the action of acid hydrolysis, enzymes, etc. these substances can be rapidly inactivated or destroyed. In addition, the size and structure of macromolecular drugs can inhibit absorption.

[6] The previously known methods of oral administration of vulnerable pharmacological agents are based on the joint administration of adjuvants (for example, resorcinol and nonionic surfactants such as polyoxyethylene oleyl ether and n-hexadecyl polyethylene ether) to artificially increase the intestinal wall permeability, as well as joint the introduction of enzyme inhibitors (for example, trypsin, diisopropyl fluorophosphate (DFF) and trasilol inhibitors) to prevent decomposition (of agents) by enzymes. Liposomes have also been described as insulin and heparin delivery systems. However, widespread use of such drug delivery systems is not possible since (1) these systems require toxic amounts of adjuvants or inhibitors; (2) no suitable loads (transported agents) with low molecular weight, i.e. active substances; (3) these systems have low stability and insufficient shelf life; (5) systems are not able to protect the active substance (load); (6) systems alter the active substance, making it harmful; or (7) systems do not allow or do not stimulate the absorption of the active substance.

[7] Protein microbeads are used for drug delivery. See, for example, US Patent Nos. 5,401,516; 5,443,841; and Re. 35,862. In addition, specifically modified amino acids are used for drug delivery. See, for example, US Pat. Nos. 5,629,020; 5,643,957; 5,766,633; 5,776,888; and 5,866,536, and the publication of international applications WO 98/49135; WO 00/06534; WO 00/07979; WO 00/40203; WO 00/47188; WO 00/50386; WO 00/59863; WO 01/32130, WO 01/32596, WO 01/44199, WO 01/51454, WO 02/02509, WO 02/15959, WO 02/16309, WO 02/20466, WO 02/19969, WO 02/69937, WO 03/45306.

[8] More recently, to obtain polymeric substances that provide delivery, the polymer was conjugated to a modified amino acid or its derivative through a linking group. The modified polymer may be any polymer, but the preferred polymers are, but are not limited to, polyethylene glycol (PEG) and its derivatives. See, for example, publication of international application WO 00/40203.

However, there is still a need for simple, cheap delivery systems that are easy to prepare and that can deliver a wide range of active substances into the body in various ways.

SUMMARY OF THE INVENTION

[9] The present invention provides compounds and compositions that facilitate the delivery of active substances. Delivery-promoting compounds (carrier compounds) provided by the present invention include the compounds shown below and their pharmaceutically acceptable salts:

wherein:

R ₁ represents - (CH ₂ ) _m —R ₈ where m = 0 or 1;

R ₂ -R ₆ are independently selected from the group consisting of hydrogen, hydroxyl, halogen, alkyl C ₁ -C _4, alkenyl C ₂ -C ₄ alkynyl, C ₂ -C _4, alkoxy C ₁ -C _4, and cyano;

R ₇ is selected from the group consisting of alkyl C ₁ -C _10, alkenyl C ₂ -C ₁₀ alkynyl and C ₂ -C _10;

R _{8 is} selected from the group consisting of cyclopentyl, cyclohexyl and phenyl, wherein when R ₈ is phenyl, m = 1; moreover

R ₈ is optionally substituted alkyl C ₁ -C _4, alkoxy C ₁ -C _4, halogen, or hydroxy, or combination thereof.

[10] In one embodiment, R ₇ is C ₁ alkyl.

[11] In another example implementation, R ₇ is C ₂ alkyl.

[12] In another example implementation, R ₇ is C ₃ alkyl.

[13] In another example implementation, R ₇ is C ₄ alkyl.

[14] In another example implementation, R ₇ is C ₅ alkyl.

[15] In another example implementation, R ₇ is C ₆ alkyl.

[16] In another example implementation, R ₇ is C ₇ alkyl.

[17] In another example implementation, R ₇ is C ₈ alkyl.

[18] Preferred compounds are, inter alia, the following compounds and their salts suitable for pharmaceutical use:

[19] To other carrier compounds proposed in

the present invention include compounds having the formula: (Compound B) and their pharmaceutically acceptable salts, wherein:

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

R ₂ -R ₆ are selected independently from the group consisting of hydrogen, hydroxyl, halogen, alkyl C ₁ -C _4, alkenyl C ₂ -C ₄ alkynyl, C ₂ -C _4, alkoxy C ₁ -C _4, and cyano , and

R _{7 is} selected from the group consisting of C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, and C ₂ -C ₁₀ alkynyl.

[20] In one embodiment, R ₂ -R ₆ are independently hydrogen, methyl, halogen, methoxy.

[21] In another example implementation, R ₂ -R ₆ are independently hydrogen, methyl, chloro, methoxy.

[22] In another example implementation, R ₂ -R ₆ are independently hydrogen, methyl, fluoro, methoxy.

[23] In another example implementation, R ₂ -R ₆ are independently hydrogen, methyl, iodo, methoxy.

[24] In another example implementation, R ₂ -R ₆ are independently hydrogen, methyl, bromo, methoxy.

[25] In another example implementation, R ₁ is C ₁ -C ₃ alkyl.

[26] In another example implementation, R ₁ is methyl.

[27] In another example implementation, R ₁ is ethyl.

[28] In another example implementation, R ₁ is isopropyl.

[29] In another example implementation, R ₂ is methyl.

[30] In another example implementation, R ₂ is halogen.

[31] In another example implementation, R ₂ is chlorine.

[32] In another example implementation, R ₂ is fluorine.

[33] In another example implementation, R ₄ is methyl.

[34] In another example implementation of R ₄ is methoxy.

[35] In another example implementation, R ₄ is halogen.

[36] In another example implementation, R ₄ is chlorine.

[37] In another example implementation, R ₄ is fluorine.

[38] In another example implementation of R ₄ is cyano.

[39] In another example implementation, R ₇ is C ₁ alkyl.

[40] In another example implementation, R ₇ is C ₂ alkyl.

[41] In another example implementation, R ₇ is C ₂ alkyl, branched methyl.

[42] In another example implementation, R ₇ is C ₃ alkyl.

[43] In another embodiment, R ₇ is C ₃ alkyl branched methyl.

[44] In another embodiment, R ₇ is C ₄ alkyl.

[45] In another embodiment, R ₇ is C ₅ alkyl.

[46] In another example implementation, R ₇ is C ₆ alkyl.

[47] In another example implementation, R ₇ is C ₇ alkyl.

[48] In another embodiment, R ₇ is C ₈ alkyl.

[49] Preferred compounds include, but are not limited to, the following compounds and their pharmaceutically acceptable salts:

Compound 35

10- [methyl (4-methylphenyl) amino] -10-oxodecanoic acid

Compound 36

3 - [(4-chlorophenyl) (methyl) amino] -3-oxopropionic acid Compound 37

3 - [(4-chlorophenyl) (ethyl) amino] -3-oxopropionic acid Compound 38

5 - [(4-chlorophenyl) (ethyl) amino] -4-methyl-5-oxopentanoic acid Compound 39

10 - [(4-chlorophenyl) (methyl) amino] -10-oxodecanoic acid Compound 40

4 - [(4-chlorophenyl) (ethyl) amino] -4-oxobutanoic acid Compound 41

5 - [(4-chlorophenyl) (methyl) amino] -5-oxopentanoic acid

Compound 42

7- [ethyl (2-methylphenyl) amino] -7-oxoheptanoic acid Compound 43

6- [ethyl (2-methylphenyl) amino] -6-oxohexanoic acid Compound 44

4-methyl-5- [methyl (4-methylphenyl) amino] -5-oxopentanoic acid Compound 45

5 - [(4-chlorophenyl) (methyl) amino] -4-methyl-5-oxopentanoic acid Compound 46

8- [methyl (2-methylphenyl) amino] -8-oxo-octanoic acid Compound 47

5- [ethyl (2-methylphenyl) amino] -3-methyl-5-oxopentanoic acid

Compound 48

5- [isopropyl (phenyl amino] -4-methyl-5-oxopentanoic acid Compound 49

5- [ethyl (2-methylphenyl) amino] -4-methyl-5-oxopentanoic acid Compound 50

4 - [(4-chlorophenyl) (methyl) amino] -3-methyl-4-oxobutanoic acid Compound 51

9- [methyl (4-methylphenyl) amino] -9-oxononanoic acid Compound 52

8- [ethyl (2-methylphenyl) amino] -8-oxo-octanoic acid Compound 53

8- [isopropyl (phenylamino] -8-oxo-octanoic acid

Compound 54

9- [methyl (2-methylphenyl) amino] -9-oxononanoic acid Compound 55

6- [ethyl (phenyl) amino] -6-oxohexanoic acid Compound 56

7- [ethyl (2-methylphenyl) amino] -7-oxoheptanoic acid Compound 57

9- [ethyl (2-methylphenyl) amino] -9-oxononanoic acid Compound 58

10- [ethyl (phenyl) amino] -10-oxodecanoic acid Compound 61

5- [ethyl (phenyl) amino] -4-methyl-5-oxopentanoic acid

Compound 62

4- [ethyl (2-methylphenyl) amino] -4-oxobutanoic acid Compound 63

5- [methyl (4-methylphenyl) amino] -5-oxopentanoic acid Compound 64

5- [methyl (2-methylphenyl) amino] -5-oxopentanoic acid Compound 65

5- [isopropyl (phenyl) amino] -5-oxopentanoic acid Compound 66

5- [ethyl (phenyl) amino] -4-methyl-5-oxopentanoic acid Compound 67

5- [ethyl (phenyl) amino] -5-oxopentanoic acid Compound 68

6- [methyl (4-methylphenyl) amino] -6-oxohexanoic acid

Compound 69

6- [methyl (2-methylphenyl) amino] -6-oxohexanoic acid Compound 70

6- [ethyl (2-methylphenyl) amino] -6-oxohexanoic acid Compound 71

7- [methyl (4-methylphenyl) amino] -7-oxoheptanoic acid Compound 72

7- [methyl (2-methylphenyl) amino] -7-oxoheptanoic acid Compound 73

5 - [(4-chlorophenyl) (ethyl) amino] -5-oxopentanoic acid Compound 74

6 - [(4-chlorophenyl) (methyl) amino] -6-oxohexanoic acid Compound 75

7 - [(4-chlorophenyl) (methyl) amino] -7-oxoheptanoic acid

Compound 76

7 - [(4-cyanophenyl) (methyl) amino] -7-oxoheptanoic acid Compound 77

7 - [(4-methoxyphenyl) (methyl) amino] -7-oxoheptanoic acid Compound 78

8 - [(4-cyanophenyl) (methyl) amino] -8-oxocaprylic acid Compound 79

8 - [(4-fluorophenyl) (methyl) amino] -8-oxo-octanoic acid Compound 80

9 - [(2-fluorophenyl) (methyl) amino] -9-oxononanoic acid Compound 81

9 - [(4-fluorophenyl) (methyl) amino] -9-oxononanoic acid Compound 82

7 - [(4-chlorophenyl) (ethyl) amino] -7-oxoheptanoic acid

Compound 83

8 - [(4-chlorophenyl) (methyl) amino] -8-oxo-octanoic acid Compound 84

9 - [(4-chlorophenyl) (methyl) amino] -9-oxononanoic acid Compound 85

9- [isopropyl (phenyl amino] -9-oxononanoic acid Compound 86

9- [ethyl (2-methylphenyl) amino] -9-oxononanoic acid Compound 87

10- [methyl (2-methylphenyl) amino] -10-oxodecanoic acid Compound 88

10- [isopropyl (phenyl) amino] -10-oxodecanoic acid Compound 89

10- [ethyl (phenyl) amino] -10-oxodecanoic acid

Compound 90

9 - [(4-cyanophenyl) (methyl) amino] -9-oxononanoic acid Compound 91

9 - [(4-methoxyphenyl) (methyl) amino] -9-oxononanoic acid Compound 92

10 - [(4-cyanophenyl) (methyl) amino] -10-oxodecanoic acid Compound 93

10 - [(4-methoxyphenyl (methyl) amino] -10-oxodecanoic acid Compound 94

10 - [(2-fluorophenyl) (methyl) amino] -10-oxodecanoic acid Compound 95

10 - [(4-fluorophenyl) (methyl) amino] -10-oxodecanoic acid Compound 96

9 - [(4-chlorophenyl) (ethyl) amino] -9-oxononanoic acid

Compound 97

10 - [(4-chlorophenyl) (methyl) amino] -10-oxodecanoic acid Compound 98

10 - [(4-chlorophenyl) (ethyl) amino] -10-oxodecanoic acid Compound 99

3-methyl-4- [methyl (4-methylphenyl) amino] -4-oxobutanoic acid Compound 100

3-methyl-4- [methyl (2-methylphenyl) amino] -4-oxobutanoic acid Compound 101

4- [isopropyl (phenylamino] -3-methyl-4-oxobutanoic acid Compound 102

3-methyl-5- [methyl (4-methylphenyl) amino] -5-oxopentanoic acid Compound 103

3-methyl-5- [methyl (2-methylphenyl) amino] -5-oxopentanoic acid

Compound 104

4- [ethyl (2-methylphenyl) amino] -3-methyl-4-oxobutanoic acid Compound 105

4-methyl-5- [methyl (4-methylphenyl) amino] -5-oxopentanoic acid Compound 106

4-methyl-5- [methyl (2-methylphenyl) amino] -5-oxopentanoic acid Compound 107

5 - [(4-chlorophenyl) (methyl) amino] -3-methyl-5-oxopentanoic acid Compound 108

4 - [(4-chlorophenyl) (ethyl) amino] -3-methyl-4-oxobutanoic acid Compound 109

5 - [(4-chlorophenyl) (methyl) amino] -4-methyl-5-oxopentanoic acid

Compound 110

5 - [(4-chlorophenyl) (ethyl) amino] -3-methyl-5-oxopentanoic acid

[50] Other carrier compounds provided by the present invention include those having the formula:

and their pharmaceutical acceptable salts, while

n = 1 to 9, and

R ₁ -R ₅ are, independently, hydrogen, alkyl C ₁ -C _4, alkoxy C ₁ -C _4, alkenyl C ₂ -C _4, halogen, hydroxyl, -NH-C (O) -CH ₃ , or —O — C ₆ H ₅ .

[51] Preferred carrier compounds are, without limitation, compounds having the following formulas and their salts:

[52] In one example implementation, n = 2-8.

[53] In another example implementation n = 8.

[54] In another example implementation n = 7.

[55] In another example implementation n = 6.

[56] In another example implementation n = 5.

[57] In another example implementation n = 4.

[58] In another example implementation n = 3.

[59] In another example implementation, n = 2 and the remaining R groups are hydrogens.

[60] In another example implementation, n = 8 and the remaining R groups are hydrogens.

[61] In another example implementation, n = 7 and the remaining R groups are hydrogens.

[62] In another example implementation, n = 6 and the remaining R groups are hydrogens.

[63] In another example implementation, n = 5 and the remaining R groups are hydrogens.

[64] In another example implementation, n = 4 and the remaining R groups are hydrogens.

[65] In another example implementation, n = 3 and the remaining R groups are hydrogens.

[66] In another example implementation, n = 2 and the remaining R groups are hydrogens.

[67] In another example implementation, R ₁ and R ₅ are hydrogens.

[68] In another example implementation, R ₁ and R ₅ are hydrogens and n = 2.

[69] In another example implementation, R ₃ is hydroxyl.

[70] In another example implementation, R ₃ is hydroxyl and N = 8.

[71] In another example implementation, R ₁ is hydroxyl.

[72] In another example implementation, R ₁ is hydroxyl and N = 8.

[73] In another example implementation of R ₃ is methoxy.

[74] In another example implementation, R ₃ is methoxy and N = 2.

[75] In another example implementation, R ₃ is methoxy and N = 3.

[76] In another example implementation, R ₂ and R ₄ are halogen and N = 2.

[77] In another example implementation, R ₂ and R ₄ are fluoro.

[78] In another example implementation, R ₂ and R ₄ are fluoro and N = 2.

[79] In another example implementation, R ₁ and R ₃ are methyl.

[80] In another example implementation, R ₁ and R ₃ are methyl and N = 2.

[81] In another example implementation, R ₂ and R ₄ are methyl, R ₃ is methoxy and N = 4.

[82] In another example implementation, R ₃ is isopropyl.

[83] In another example implementation, R ₃ is isopropyl and N = 3.

[84] In another example implementation, R ₁ is methoxy.

[85] In another example implementation, R ₁ is methoxy and N = 2.

[86] In another example implementation, R ₃ is halogen.

[87] In another example implementation, R ₃ is halogen and N = 2.

[88] In another example implementation, R ₃ is fluorine and N = 2.

[89] In another example implementation of R ₃ is methoxy.

[90] In another example implementation, R ₃ is methoxy and N = 4.

[91] In another example implementation, R ₂ and R ₄ are methyl.

[92] In another example implementation, R ₂ and R ₄ are methyl and N = 2.

[93] In another example implementation, R ₂ and R ₄ are methyl and N = 4.

[94] In another example implementation, R ₂ and R4 are methyl and N = 6.

[95] In another example implementation, R ₂ and R ₃ are methyl and N = 4.

[96] In another example implementation, R ₂ and R ₃ are methyl and N = 2.

[97] In another example implementation, R ₁ and R ₄ are methyl and N = 2.

[98] In another example implementation, R ₁ and R ₄ are halogen.

[99] In another example implementation, R ₁ and R ₄ are halogen and N = 2.

[100] In another example implementation, R ₁ and R ₄ are halogen and N = 4.

[101] In another example implementation, R ₁ and R ₄ are chlorine.

[102] In another example implementation, R ₁ and R ₄ are chlorine and N = 2.

[103] In another example implementation, R ₁ and R ₄ are chlorine and N = 4.

[104] In another example implementation, R ₁ and R ₄ are hydroxyls.

[105] In another example implementation, R ₁ and R ₄ are hydroxyls and N = 8.

[106] In one example implementation of 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.

[107] Preferred compounds include, without limitation, the compounds shown below.

[108] Other compounds of the carrier substances proposed in the present invention include compounds having the formula:

and their pharmaceutically acceptable salts, while

R ₁ -R ₄ are, independently of one another, hydrogen, C ₁ -C ₄ alkyl, C ₂ -C ₄ alkenyl, halogen, C ₁ -C ₄ alkoxy, or hydroxyl.

[109] In one embodiment, R1-R4 are, independently of each other, hydrogen, methyl, methoxy, halogen, or isopropyl.

[110] In one example implementation of R1-R4, all are hydrogens.

[111] In another embodiment, R2 and R4 are halogens, preferably bromine or preferably chlorine, or preferably iodine, or preferably fluorine.

[112] In another embodiment, R2 and R4 are halogens, preferably bromine or preferably chlorine, or preferably iodine, and R1 and R3 are hydrogens.

[113] In another preferred embodiment, R2 and R4 are isopropyls.

[114] In another preferred embodiment, R2 and R4 are isopropyls, and R1 and R3 are hydrogens.

[115] In another preferred embodiment, R4 is methyl.

[116] In another preferred embodiment, R4 is methyl and R1-R3 are hydrogens.

[117] In another preferred embodiment, R3 is halogen, preferably chlorine.

[118] In another preferred embodiment, R3 is halogen, preferably chlorine, and R1, R2 and R4 are hydrogens.

[119] In another preferred embodiment, R3 is methoxy.

[120] In another preferred embodiment, R3 is methoxy, and R1, R2 and R4 are hydrogens.

[121] In another preferred embodiment, R2 is halogen, preferably bromine.

[122] In another preferred embodiment, R2 is halogen, preferably bromine, and R1, R2 and R4 are hydrogens.

[123] In another preferred embodiment, R2 is halogen, preferably chlorine.

[124] In another preferred embodiment, R2 is halogen, preferably chlorine, and R1, R3 and R4 are hydrogens.

[125] In another preferred embodiment, R2 is methoxy.

[126] In another preferred embodiment, R2 is methoxy, and R1, R3 and R4 are hydrogens.

[127] In another preferred embodiment, R2 is methyl.

[128] In another preferred embodiment, R2 is methyl, and R1, R3 and R4 are hydrogens.

[129] Preferred carrier compounds include, but are not limited to, compounds having the following formulas and their salts:

Connection140 3- (2-hydroxy-benzoylamino) -butyric acid

Compound 141 3- (3,5-dibromo-2-hydroxy-benzoylamino) butyric acid

Compound 142 3- (3,5-dichloro-2-hydroxy-benzoylamino) butyric acid

Compound 143 3- (2-hydroxy-3,5-diiodo-benzoylamino) butyric acid

Compound 144 3- (2-hydroxy-3-methyl-benzoylamino) butyric acid

Compound 145 3- (4-chloro-2-hydroxy-benzoylamino) butyric acid

Compound 146 3- (2-hydroxy-4-methoxy-benzoylamino) butyric acid

Compound 147 3- (5-bromo-2-hydroxy-benzoylamino) butyric acid

Compound 148 3- (5-chloro-2-hydroxy-benzoylamino) butyric acid

Compound 149 3- (2-hydroxy-5-methoxy-benzoylamino) butyric acid

Compound 150 3- (2-hydroxy-5-methyl-benzoylamino) butyric acid

Compound 151 3- (2-hydroxy-3,5-diisopropyl-benzoylamino) butyric acid

[130] Other carrier compounds provided by the present invention include those having the formula:

and their pharmaceutically acceptable salts, with:

one of the groups R1-R5 has the general structure - (CH ₂ ) _n -COOH,

where n = 0-6;

the remaining four members of R1-R5 are independently of each other hydrogen, C ₁ -C ₄ alkyl, C ₂ -C ₄ alkenyl, halogen, C ₁ -C ₄ alkoxy, or hydroxyl; and

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

[131] In one example implementation, n = 0-4.

[132] In another example implementation, n = 0.

[133] In another example implementation, n = 1.

[134] In another example implementation, R1-R10 are preferably, independently of each other, hydrogen, halogen, methyl and methoxy.

[135] In another embodiment, R1-R10 are preferably, independently of each other, chlorine, halogen, methyl, and methoxy.

[136] In another example implementation, when the general structure is (CH ₂ ) _n -COOH, attached to R1, the remaining R groups are hydrogens.

[137] In another example implementation, when the general structure is (CH ₂ ) _n -COOH is attached to R1, the remaining R groups are hydrogens and n = 0.

[138] In another example implementation, when the general structure is (CH ₂ ) _n -COOH is attached to R1, the remaining R groups are hydrogens and n = 1.

[139] In another example implementation, when the general structure is (CH ₂ ) _n -COOH, attached to R3, the remaining R groups are hydrogens.

[140] In another example implementation, when the general structure is (CH ₂ ) _n -COOH is attached to R3, the remaining R groups are hydrogens, and n = 0.

[141] In another example implementation, when the general structure is (CH ₂ ) _n -COOH, attached to R3, the remaining R groups are hydrogens.

[142] In another example implementation, when the general structure is (CH ₂ ) _n -COOH is attached to R3, the remaining R groups are hydrogens and n = 1.

[143] In another example implementation, R5 is methoxy when the general structure of - (CH ₂ ) _n -COOH is attached to R2.

[144] In another example implementation, R5 is methoxy when the general structure of - (CH ₂ ) _n -COOH is attached to R2, the remaining R groups are hydrogens.

[145] In another example implementation, R5 is methoxy when the general structure - (CH ₂ ) _n -COOH is attached to R2, and n = 0.

[146] In another example implementation, R5 is methoxy when the general structure of - (CH ₂ ) _n -COOH is attached to R2, and n = 0, the remaining R groups are hydrogens.

[147] In another example implementation, R1 and R5 are methyl when the general structure - (CH ₂ ) _n —COOH is attached to R3.

[148] In another example implementation, R1 and R5 are methyl, when the general structure - (CH ₂ ) _n —COOH is attached to R3, the remaining R groups are hydrogens.

[149] In another example implementation, R1 and R5 are methyl when the general structure - (CH ₂ ) _n —COOH is attached to R3 and n = 0.

[150] In another example implementation, R1 and R5 are methyl, when the general structure - (CH ₂ ) _n -COOH is attached to R3 and n = 0, the remaining R groups are hydrogens.

[151] In another example implementation, R1 or R5 is methoxy when the general structure - (CH ₂ ) _n —COOH is attached to R3 and n = 0.

[152] In another example implementation, R1 or R5 is methoxy when the general structure - (CH ₂ ) _n —COOH is attached to R3 and n = 0, the remaining R groups are hydrogens.

[153] In another embodiment, R2 or R4 is halogen, preferably chlorine, when the general structure of - (CH ₂ ) _n -COOH is attached to R3.

[154] In another embodiment, R2 or R4 is halogen, preferably chlorine, when the general structure of - (CH ₂ ) _n -COOH is attached to R3, the remaining R groups are hydrogens.

[155] In another example implementation, R2 or R4 is halogen, preferably chlorine, when the general structure - (CH ₂ ) _n —COOH is attached to R3 and n = 0.

[156] In another embodiment, R2 or R4 is halogen, preferably chlorine, when the general structure of - (CH ₂ ) _n -COOH is attached to R3 and n = 0, the remaining R groups are hydrogens.

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

[158] Other carrier compounds for the delivery of substances proposed in the present invention include compounds of the formula:

and their pharmaceutically acceptable salts, with:

n is from 1 to 9; and

R ₁ -R ₉ are, independently of one another, hydrogen, C ₁ -C ₄ alkyl, C ₂ -C ₄ alkenyl, halogen, C ₁ -C ₄ alkoxy, or hydroxyl.

[159] According to one preferred embodiment, n = 3-7, preferably in one preferred embodiment, n = 3, preferably in another preferred embodiment, n = 4; preferably, in another preferred embodiment, n = 5; preferably, in another preferred embodiment, n = 6; preferably, in another preferred embodiment, n = 7.

[160] According to another preferred example, R1-R8 is hydrogen.

[161] According to another preferred example, R3 is halogen, preferably in one embodiment, R3 is chlorine, preferably in another embodiment, R3 is bromine.

[162] According to another preferred example, R2 is methoxy.

[163] According to another preferred example, R2 is methyl.

[164] According to another preferred example, R3 is methoxy.

[165] According to another preferred example, R3 is methyl.

[166] According to another preferred example, R6 is methoxy.

[167] According to another preferred example, R9 is hydrogen.

[168] According to another preferred example, R9 is hydroxyl.

[169] According to another preferred example, R9 is halogen, preferably in one embodiment of chlorine.

[170] According to another preferred example, R3 and R6 are both methoxy.

[171] According to another preferred example, R3 and R6 are both methoxy and the remaining R groups are hydrogen.

[172] According to another preferred example, R2 is methyl and R3 is chlorine.

[173] According to another preferred example, R2 is methyl and R3 is chlorine and the remaining R groups are hydrogens.

[174] According to another preferred example, R2 is methyl and R9 is chlorine.

[175] According to another preferred example, R2 is methyl and R9 is chlorine and the remaining R groups are hydrogens.

[176] According to another preferred example, R3 is methyl and R9 is chlorine.

[177] According to another preferred example, R3 is methyl and R9 is chlorine and the remaining R groups are hydrogens.

[178] Preferred carrier compounds for the delivery of substances include, without limitation, compounds having the following formulas and their salts:

Compound 165 4- (2- (2-hydroxybenzoyl) -5-methoxyphenoxy) butyric acid

[179] Other delivery carrier compounds provided by the present invention include those having the formula:

and their pharmaceutically acceptable salts, with:

R1-R5 are, independently of one another, 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 = 1-12);

at least one of the R1-R5 groups has a common structure

-O- (CH ₂ ) _n -COOH

where n = 1-12; and

R6-R10 are, independently of one another, hydrogen, C ₁ -C ₄ alkyl, C ₂ -C ₄ alkenyl, halogen, C ₁ -C ₄ alkoxy, or hydroxyl.

[180] Preferably, only one of the R1-R5 groups has the formula - (CH ₂ ) _n —COOH. In other words, the four members of R1-R5 are, independently of one another, hydrogen, C ₁ -C ₄ alkyl, C ₂ -C ₄ alkenyl, halogen, C ₁ -C ₄ alkoxy, or hydroxyl, and the remaining member R1-R5 is —O— (CH ₂ ) _n —COOH (where n = 1-12).

[181] In one preferred embodiment, n = 1-12.

[182] In another preferred embodiment, n = 1-10.

[183] In another preferred embodiment, n = 1-6.

[184] In another preferred embodiment, n = 1-4.

[185] In another preferred embodiment, n = 10.

[186] In another preferred embodiment, n = 4.

[187] In another preferred embodiment, n = 1.

[188] When the general structure - (CH ₂ ) _n —COOH is attached to R1, all other R groups are hydrogens.

[189] When the general structure - (CH ₂ ) _n —COOH is attached to R1, all other R groups are hydrogens and n = 3.

[190] When the general structure - (CH ₂ ) _n —COOH is attached to R3, all other R groups are hydrogens.

[191] When the general structure - (CH ₂ ) _n -COOH is attached to R3, all other R groups are hydrogens and n = 1.

[192] When the general structure - (CH ₂ ) _n -COOH is attached to R3, all other R groups are hydrogens and n = 4.

[193] When the general structure - (CH ₂ ) _n -COOH is attached to R3, all other R groups are hydrogens and n = 10.

[194] Preferred compounds include, but are not limited to, the following compounds and their pharmaceutically acceptable salts:

Compound 175 4- (2-Benzyloxy-phenoxy) -butyric acid

Compound 176 (4-Benzyloxy-phenoxy) acetic acid

Compound 177 11- (2-Benzyloxyphenoxy) undecanoic acid

Compound 178 5- (4-Benzyloxy-phenoxy) pentanoic acid

[195] Mixtures of these carrier compounds may also be used.

[196] The invention also provides a composition comprising a carrier compound of the present invention and at least one active substance. These formulations deliver active substances to selected biological systems, with the bioavailability of the active substances increasing or improving compared to the administration of the active substances without carrier compounds.

[197] Unit dosage forms containing these formulations are also provided. Unit doses may be in liquid or solid form, for example tablets, capsules or particles, including powder or sachets.

[198] Another embodiment is a method of introducing an active substance into an animal organism by administering a composition comprising at least one of the carrier compounds of the present invention and the active substance. Methods of administration include oral administration, administration to the colon, and administration through the lungs.

[199] Another embodiment is a method of treating a disease or achieving a desired physiological effect in an animal organism by administering a composition of the present invention.

[200] Another example of implementation is the introduction of the composition proposed in the present invention into the body of an animal to which this composition will bring a beneficial effect, and / or to an animal that needs an active substance.

[201] Another example implementation is a method of preparing a composition proposed in the present invention, by mixing at least one carrier compound proposed in the present invention, and at least one active substance.

DETAILED DESCRIPTION OF THE INVENTION

Definitions

[202] Used here and in the accompanying claims, the singular forms include numerous objects, unless the content clearly indicates otherwise. Thus, for example, the term “molecule” means one or more such molecules, the term “reagent” means one or more of these different reagents, the term “method” includes equivalent steps or methods known to a typical person skilled in the art, which can be modified replaced by the methods described here.

[203] The term "polyform" refers to various crystallographic forms of a substance.

[204] The term "hydrate" as used herein includes, without limitation, (i) a substance containing water and combined with it in a molecular form; and (ii) a crystalline substance containing one or more water molecules included in the crystal lattice, or a crystalline substance containing free water

[205] As used herein, the term “solvate” includes, without limitation, a molecular or ionic complex of molecules or ions of a solvent with molecules or ions of a carrier substance.

[206] The term “carrier substance” refers to any carrier compounds disclosed or incorporated herein by reference, including pharmaceutically acceptable salts thereof.

[207] An “effective amount of a pharmaceutical composition” is an amount of a pharmaceutical composition described that is effective in treating or preventing the condition of a subject to which it is administered over a period of time, for example, provides a therapeutic effect during a desired dosage interval.

[208] The term “treat,” “treat,” or “treated” refers to prophylactic prevention, recovery, healing, alleviation, mitigation, alteration, cure, improvement, or effect on a condition (eg, disease), symptoms of a condition, or predisposition to state.

[209] An “effective amount of a delivery substance” is an amount of a carrier substance that promotes absorption of the required amount of active substance.

[210] The term “subject” includes mammals such as rodents, cows, pigs, dogs, cats, primates, and in particular humans.

[211] As used herein, the term "PEP" means the area under the curve "plasma concentration - time", which is calculated by the trapezoidal formula throughout the dosing interval, for example, a 24-hour interval.

[212] The term "mean" before a pharmacokinetic value (eg, average peak) is the arithmetic mean of the pharmacokinetic parameter, unless otherwise indicated.

[213] As used herein, the term “approximately” means within 10% of a given value, preferably within 5%, and more preferably within 1% of that value. Alternatively, the term “approximately” means that the value can be reduced within the scientifically acceptable margin of error for this type of value, which depends on how well you can measure using the available tools.

[214] “Indication” means a use in which a medicine is administered either to prevent or treat a condition, and may be used interchangeably with the terms “treatment”, “treated” or “treating”.

[215] The term "substituted" as used herein includes, without limitation, substitution with any one or any combination of the following substituents: halogens, hydroxide, alkyl C ₁ -C ₄ alkoxy and C ₁ -C _4.

[216] The terms "alkyl", "alkoxy", "alkylene", "alkenylene", "alkyl (arylene)", and "aryl (alkylene)" include, without limitation, linear and branched alkyl, alkoxy, alkylene, alkenylene, alkyl (arylene), and aryl (alkylene) groups, respectively.

[217] The term "peptide YY" or "PYY" refers to a polypeptide of the YY peptide obtained or extracted from any species. Thus, the term “PYY” includes as a human full-length peptide containing 36 amino acids, as set forth in SEQ ID NO: in International Publication No. WO 02/47712 (which is a PCT duplicate of US Patent Publication No. 2002/0141985, incorporated herein by reference) and Tatemoto, Proc Natl Acad Sci U.S.A. 79: 2514-8, 1982, as well as species variations of PYY, including, for example, PUCs of mice, hamsters, chickens, cows, rats and dogs. The term "PYY agonist" means any compound that causes an effect similar to the action of PYY, which reduces the availability of nutrients, for example, compound (1), which is active in food intake, gastric emptying, pancreatic secretion, or weight loss test, as described in the Examples 1, 2, 5, or 6 in WO 02/47712 and in U.S. Patent Publication No. 2002/0141985, and (2) which, upon analysis, specifically binds the Y receptor (Example 10 in WO 02/47712 and US Patent Publication No. 2002/0141985) or in the analysis of competitive binding to labeled PYY or PYY [3-36] from certain tissues containing an excess of Y receptors, including, for example, from the area of postrem (Example 9 in WO 02/47712 and in Publication of US patents No. 2002/0141985), while the agonist PYY is not a pancreatic polypeptide. It is preferred that the PYY agonists bind in such assay methods with an affinity greater than about 1 μM, and more preferably an affinity greater than about 1 to about 5 μM.

[218] Such agonists may comprise a polypeptide having a functional domain of PYY, an active fragment of PYY, a chemical reagent, or a small molecule. PYY agonists can be peptide or non-peptide compounds and include "PYY agonist analogs", which include any compound structurally similar to PYY and having PYY activity, typically due to binding or other method of direct or indirect interaction with the PYY receptor or other receptor or receptors with which PYY itself can interact, causing a biological response. Such compounds include PYY derivatives, PYY fragments, extended PYY molecules containing more than 36 amino acids, truncated PYY molecules containing less than 36 amino acids, and substituted PYY molecules containing one or more different amino acids, or any combination of the above compounds. These compounds can also be modified using methods such as pegylation, amidation, glycosylation, acylation, sulfation, phosphorylation, acetylation and cyclization.

[219] One such analogue of a PYY agonist is PYY [3-36], designated SEQ ID NO: in WO 02/47712 and US 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 parentheses refer to truncated polypeptides having a full length peptide sequence at all amino acid positions shown in parentheses. Thus, PYY [3-36] has a sequence identical to PYY for all amino acids from 3 to 36. Immunoreactivity of POC [3-36] is approximately 40% of the total peptide YY-like immunoreactivity in intestinal extracts of humans and dogs and from approximately 36 % of the total immunoreactivity of the YY peptide in plasma when the subject is hungry, to slightly above 50% in the satiated state. PYY [3-36] apparently represents the cleavage product of the YY peptide with dipeptidyl peptidase-IV (DPP4). The YY peptide [3-36] is reported to be a selective ligand at the Y2 and Y5 receptors, which appear to be pharmacologically unique in Y-truncated analogs of neuropeptides, preferably with N atoms at the ends (for example, C terminal fragments). A PYY agonist can bind to a PYY receptor with higher or lower affinity, while it exhibits a longer or shorter half-life in vivo or in vitro, or is more or less effective than natural DYY.

[220] Other suitable PYY agonists include those described in International Publication No. WO 98/20885, which is incorporated by reference.

[221] As used herein, the term “heparin” refers to all forms of heparin, including, without limitation, unfractionated heparin, heparinoids, dermatans, chondroitins, low molecular weight heparin (eg, tinzaparin (including sodium tinzaparin)), very low molecular weight heparin weight, and ultra-low molecular weight heparin. Non-limiting examples include unfractionated heparin, e.g., sodium heparin (e.g., USP heparin, available from Scientific Protein Labs of Waunakee, Wisconsin). In general, heparin has a molecular weight of from 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 heparin has a molecular weight of between about 3000 and about 9000 daltons. Non-limiting examples of low molecular weight heparin include tinzaparin, enoxaprine, and daltiparin. Tinzaparin has been approved by the Food and Drug Administration (FDA US, USA) for the treatment of acute symptomatic deep vein thrombosis with or without pulmonary embolism when administered with sodium warfarin. Pharmion Corporation ™ tinazaparin sodium is available under the brand name Innohep of Boulder, Colorado. The term “very low molecular weight heparin” generally refers to heparin in which at least 80% (by weight) of heparin has a molecular weight of between about 1,500 and about 5,000 daltons. A non-limiting example of 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 heparin has a molecular weight of between about 1000 and about 2000 daltons. A non-limiting example of ultra-low molecular weight heparin is fondiparinax.

Carrier Reagents

[222] The carrier reagents of the present invention may be in the form of a free acid or a pharmaceutically acceptable salt. Suitable pharmaceutically acceptable salts include, but are not limited to, organic and inorganic salts, for example, alkali metal salts such as sodium, potassium, and lithium; alkaline earth metal salts such as magnesium, calcium and barium; ammonium salts; basic amino acids such as lysine and arginine; and organic amines such as dimethylamine or pyridine. In one embodiment, the salts are sodium salts. Salts may be mono- or polyvalent salts, such as monosodium salts and disodium salts. Salts may also be solvates, including ethanol solvates and hydrates. Examples of pharmaceutically acceptable salts include, but are not limited to, sodium salts, hydrochloric acid, sulfuric acid, phosphoric acid, citric acid, acetic acid, sulfates, phosphates, chlorides, bromides, iodides, acetates, propionates, hydrobromic acid, sodium hydroxide, potassium hydroxide, ammonium hydroxide; and potassium carbonate. These salts can be prepared by methods known in the art. For example, sodium salts can be prepared by dissolving the carrier reagent in ethanol and adding aqueous sodium hydroxide. The carrier reagent can be purified by recrystallization or fractionation on one or more solid chromatographic carriers, single or in series. Suitable solvents for recrystallization include, without limitation, acetonitrile, methanol and tetrahydrofuran. Fractionation can be carried out (i) on a suitable chromatographic support, such as alumina, using methanol / n-propanol mixtures as the mobile phase, (ii) using reverse phase chromatography using trifluoroacetic acid / acetonitrile mixtures as the mobile phase, or ( iii) by ion exchange chromatography using water or a suitable buffer as the mobile phase. When using anion exchange chromatography, it is preferred that the gradient of sodium chloride is 0-500 mM.

[224] The carrier reagent may comprise a polymer conjugated to it using 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 carrier polymer reagent is not a polypeptide or polyamino acid. The polymer may be any polymer, including, without limitation, alternating copolymers, block copolymers and random copolymers that are safe for mammals. Preferred polymers include, without limitation, polyethylene; polyacrylates; polymethacrylates; poly (hydroxyethylene); polypropylene); polypropylene glycol; polyethylene glycol (PEG); and their derivatives and combinations. The molecular weight of the polymer generally ranges from about 100 to about 200,000 daltons. Preferably, the molecular weight of the polymer is in the range of from about 200 to about 10,000 daltons. In one embodiment, the molecular weight of the polymer is in the range of from about 200 to about 600 daltons, and more preferably lies in the range of from about 300 to about 550 daltons. U.S. Patent No. 6,627,228 is framed here by full reference.

[225] The amount of carrier reagent in a solid pharmaceutical composition is an effective amount and can be determined for a particular composition by methods known to one skilled in the art. In the General case, the weight ratio of the carrier reagent to the active substance is in the range from about 0.1: 1 to about 1000: 1. The weight ratio will vary depending on the active substance and the specific indication according to which the active substance is introduced into the body.

[226] Other suitable carrier reagents of the present invention are described in US Pat. Nos. 6,846,844, 6,699,467, 6,693,208, 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, 6,331,318, 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, 5,866,510, 5,95, 5,85, 5,95, 5,85, 5,85, 5,85, 5,85, 5, 5, 9, 5, 5, 5, 9, 5, 5, 5, 10, 5, 9 The carrier reagents of the present invention are also described in US Patent Publication Nos. 20050009748, 20040110839, 20040106825, 20040068013, 20040062773, 20040022856, 20030235612, 20030232085, 20030225300, 20030198658, 20030133953, 20030078302, 20030072740, 20030045579, 20030012817, 20030008900, 20020001022000020102 20020013497, 20020001591, 20010039258, and 20010003001. The carrier reagents of the present invention are also described in International Publications Nos. WO 2005/020925, WO 2004/104018, WO 2004/080401, WO 2004/062587, WO 2003/057650, WO 2003/057170, WO 2003/045331, WO 2003/045306, WO 2003/026582, WO 2002/100338, WO 2002/070438, WO 2002/069937, WO 02/20466, WO 02/19969, WO 02/16309, WO 02/15959, WO 02/02509, WO 01/92206, WO 01/70219, WO 01/51454, WO 01/44199, WO 01/34114, WO 01/32596, WO 01/32130, WO 00/07979, WO 00/59863, WO 00/50386, WO 00/47188, WO 00/40203, and WO 96/30036 . Each of the above US patents, published US applications, and published international applications are hereby incorporated by reference. Said carrier reagents can be prepared by methods known in the art, as described, for example, in the aforementioned patents and published patent applications. For example, SNAC can be prepared by methods known in the art, which, for example, are described in US Pat. Nos. 5,650,386 and 5,866,536; and U.S. Patent Publications No. 2002/0065255.

Active substances

[227] Active substances suitable for use in the present invention include biologically active substances and chemically active substances, including, without limitation, pesticides, pharmacological preparations and therapeutic agents. Suitable active substances include substances that become less effective, ineffective or break down in the gastrointestinal tract, including under the influence of acid hydrolysis, enzymes, and the like. Suitable active substances also include those macromolecular substances whose physiochemical characteristics, including size, structure or charge, interfere with or delay the absorption of the active substances when taken orally.

[228] For example, a substance that must enter the body or which can beneficially influence due to improved pharmacokinetics, including delivery, for example, when oral bioavailability is limited or not significant. These substances, which are biologically or chemically active substances suitable for use in the present invention, include, without limitation, macromolecules, such as peptides, including proteins and polypeptides, including dipeptides; hormones and saccharides, 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; and, in particular, compounds that do not have oral bioavailability or with limited oral bioavailability, including those that themselves do not penetrate (or in which only part of the accepted dose penetrates) through the mucous membrane of the gastrointestinal tract and / or are susceptible to chemical decomposition by acids and enzymes in the gastrointestinal tract; or any combination thereof.

[229] Additional examples are, without limitation, the following substances, including their synthetic, natural or recombinant sources:

Amylin and amylin agonists; Adrenocorticotropin; Antigens;

Antimicrobial drugs, including antibiotics, antibacterial drugs and antifungal drugs; non-limiting examples of antibiotics include gram-positive drugs, bacteriocidal, lipopeptide and cyclic peptide antibiotics such as daptomusin and its analogues; Anti-migraine drugs, such as BIBM-4096BS and other protein antagonists genetically related to calcitonin, sumatriptan succinate; Antiviral drugs, including acyclovir, valaciclovir; Atrial natriuretic factor; Bisphosphonates, including alendronate, clodronate, etidronate, ibandronate, incadronate, minodronate, neridronate, olpadronate, pamidronate, risedronate, tiludronate, zoledronate, EB1053, and YH529; Calcitonin, including salmon, sea eel, pig, and man; Cholecystokinin (CCK) and CCK agonists, including CCK-8; Cromolin-sodium (sodium or disodium chromoglycan); Cyclosporin; Desferrioxamine (DFO); Erythropoietin; Exedin and exedin agonists, including exedin-3, exedin-4; Filgrastim Follitropin (recombinant and natural); (follicle-stimulating hormone) Glucagon-like peptide 1 (GLP-1), glucagon and glucagon-like peptide 2 (GLP-2); Glucocerebrosidase; Gonadoliberin;

Factor that stimulates the release of growth hormone; Hormones that stimulate the release of growth hormone; Growth hormones, including human growth hormones (hGH), recombinant human growth hormones (hGH), bovine growth hormones and pork growth hormones; Heparin, including unfractionated heparin, heparinoids, dermatans, chondroitins, low molecular weight heparin, very low molecular weight heparin, ultra low molecular weight heparin and synthetic heparins, including fondiparinax; Recombinant insulin, including pork, cow, human, and human, optionally containing counterions, including zinc, sodium, calcium, and ammonium; Insulin-like growth factor, including IGF-1; Interferons, including α (e.g., interferon alfacon-1 (available as Infergen® from Intermune, Inc. Brisbane, California)), β, OMEGA and γ; Interleukin -1; interleukin -2; interleukin -11; interleukin -21; Leutinizing hormone and hormone releasing hormone; Leptin (0 V protein); Monoclonal antibodies, including retuxin, TNF-alpha soluble receptors; Oxytocin; Parathyroid hormone (PTH), including its fragments, including PTH 1-34 and PTH 1-38; YY peptide (PYY), including PYY agonists, fragment 3-36;

Prostaglandins; Protease inhibitors; Somatostatin; Thrombopoietin; Vancomycin; Vasopressin; Vitamins Vaccines, including vaccines against anthrax or plague, flu and herpes;

[230] including secretion stimulants, analogues, fragments, mimetics or polyethylene glycol (PEG) -modified derivatives of these compounds; or any combination thereof.

Delivery systems

[231] The composition of the present invention comprises one or more carrier compounds of the present invention and one or more active substances. In one embodiment, one or more carrier compounds or salts of these compounds, or polyamino acids or peptides from which these compounds or salts form one or more blocks, can be used as carrier reagents by mixing with the active substance before administration to formulate introduction.

[232] Formulations for administration may be in the form of a liquid. The solvent medium may be water (e.g., for salmon calcitonin, parathyroid hormone and erythropoietin), 25% aqueous propylene glycol (e.g., for heparin), and phosphate buffer (e.g., for rhGH growth). Other dosage carriers contain polyethylene glycol. Dosage solutions can be prepared by mixing a solution of the carrier compound with a solution of the active substance immediately prior to administration. In another embodiment, a solution of the carrier compound (or active substance) can be mixed with the solid form of the active substance (or carrier compound). The carrier compound and the active substance can also be mixed in the form of dry powders. The carrier compound and the active substance can also be mixed during the process.

[233] Dosing solutions may optionally contain additives such as phosphate buffering salts, citric acid, glycols, or other dispersing agents. Stabilizing additives may be added to the solution, preferably with a concentration in the range between about 0.1 and 20% (w / v).

[234] Alternatively, the administration formulations may be in solid form, for example tablets, capsules or particles, for example powder or sachets. Solid dosage forms can be prepared by mixing the solid form of the compound with the solid form of the active substance. In another embodiment, a solid can be obtained from a solution of the compound and the active substance by methods known in the art, such as freeze-drying (lyophilization), precipitation, crystallization, and solid phase dispersion.

[235] Formulations for administration provided by the present invention may also contain one or more enzyme inhibitors. Such enzyme inhibitors include, without limitation, compounds such as actinonin or epiactinonin and their derivatives. Other enzyme inhibitors include, but are not limited to, aprotinin (Trasilol) and a Bowman-Birk inhibitor.

[236] The amount of active substance used in the composition for administration proposed in the present invention is an effective amount to achieve the goal for which a particular active substance was assigned to the patient. In general, the amount of active substance in the composition is a pharmacologically, biologically, therapeutically or chemically effective amount. However, this amount may be less than in the case of a composition used in a unit dosage form, since such a form may contain many carrier-compound / active substance formulations or may contain a pharmacologically, biologically, therapeutically or chemically effective amount divided into several doses. Then the total effective amount can be entered in cumulative doses containing, in total, an effective amount of the active substance.

[237] The total amount of active substance to be used can be determined by methods known to one skilled in the art. However, since the formulations of the present invention can transfer active substances more efficiently than formulations containing one active substance, lower amounts of biologically or chemically active substances can be administered to the subject than the amounts used in the previous unit dosage forms or delivery systems, in doing so, the same blood levels and / or therapeutic effects are achieved.

[238] The carrier compounds described in the present invention facilitate the delivery of biologically and chemically active substances, in particular upon oral, intranasal, sublingual, gastric, intestinal (intestinal) administration, including intraduodenal administration, administration through the small intestine and ileum the intestine, with subcutaneous, oral administration, administration through the colon, rectum, vaginal, intramuscular and ophthalmic administration systems, administration through the mucous membrane, and also when overcoming the blood-brain phallic barrier.

[239] The unit dosage forms may also contain any one or a combination of fillers, diluents, disintegrants, lubricants, plasticizers, colorants, flavors, flavors, sugars, sugar substitutes, salts and metered fillers, including, without limitation, water, 1,2-propanediol, ethanol, olive oil, or a combination thereof.

[240] The compounds and compositions proposed in the content of the invention are applicable for the introduction of biologically and chemically active substances into the body of any animal, including, without limitation, birds, such as chickens; mammals such as rodents, cows, pigs, dogs, cats, primates, and in particular humans; and insects.

[241] This system is particularly effective for delivering chemically or biologically active substances that would otherwise be destroyed or become less effective due to the conditions in which they fall before reaching their target area (ie, the area in which the active substance, contained in the transporting composition must be released) inside the body of the animal into which they are introduced. In particular, the compounds and compositions of the present invention are useful for the oral administration of active substances, especially those that are not usually transported by the oral route or for which it is desirable to use an improved delivery method.

[242] Compositions containing compounds and active substances are useful in delivering active substances to selected biological systems, since they increase or improve the bioavailability of active substances compared to their administration without a carrier reagent. Delivery can be improved by delivering a more active substance over a period of time, or by delivering the active substance in a specific time period (for example, delivering faster or slower delivery), or by delivering the active substance in a certain time, or within a certain period of time (for example , long delivery).

[243] Another example of the implementation of the present invention is a method of treating or preventing a disease or achieving the desired physiological effect, such as, for example, the methods listed in the table below, in animals with the introduction of the composition proposed in the present invention. It is preferable to administer an effective amount of the composition to treat or prevent a disease or achieve the desired physiological effect. Specific indications of the use of active substances can be found in (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 substances presented in the table below include their analogues, fragments, mimetics and polyethylene glycol-modified derivatives.

Active substances Disease and physiological effect Amylin and Amylin Agonists Obesity

Active substances Disease and physiological effect Adrenocorticotropin High cholesterol (to lower cholesterol) Antigens Infections Antiviral drugs, including acyclovir, valaciclovir Viral infection, including herpes simplex type I and type II Growth hormones (including recombinant human growth hormones and factors stimulating the release of growth hormone and their analogues) Growth disorders Interferons, including α, β, and γ Viral infection, including chronic cancer, hepatitis, and multiple sclerosis; Interleukins (e.g., interleukin-1; interleukin-2, interleukin-11 and 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; blood coagulation warning Calcitonin, including salmon, conger eel, pig, and human Osteoporosis; bone disease bone pain analgesic (including, relieving pain associated with osteoporosis or cancer)

Active substances Disease and physiological effect Cholecystokinin (CCK) and CCK agonists, including CCK-8 Obesity Erythropoietin Anemia; HIV / HIV therapy to treat anemia; chemotherapeutic anemia Atrial natriuretic factor Vasodilation Antigens Infections R-1- [6- [R-2-carboxy-pyrrolidin-1-yl] -6-oxo-hexanoyl] pyrrolidine-2-carboxylic acid (CPHRS) A decrease in amyloid deposits and systemic amyloidosis, often (but not always) due to Alzheimer's disease, type II diabetes, and other amyloid-related diseases Monoclonal antibodies, including retuxin, TNF-alpha soluble receptors; To prevent transplant rejection; cancer; use in tests to detect diseases Leptin (OB protein) Obesity

Active substances Disease and physiological effect Somatostatin / Ostreotide Bleeding ulcer; erosive gastritis; bleeding wind rash; diarrhea; acromegaly; pituitary adenomas secreting thyrotropin; pancreatic secretions; carcinoid syndrome; reduction of ophthalmopathy associated with proptosis / thyroid gland; spotted edema reduction / retinal degeneration Protease inhibitors HIV infection / AIDS Adrenocorticotropin High cholesterol (to lower cholesterol) Gonadoliberin Ovulation dysfunction (to stimulate ovulation) Oxytocin Violation of the body during labor (to stimulate muscle contraction) Hormone secreting hormone; follitropin To regulate reproductive function Glucocerebrosidase Gaucher disease (for the absorption of lipoprotein) Thrombopoietin Thrombocytopenia

Active substances Disease and physiological effect Filgrastim (granulocyte colony stimulating factor); GM-CSF, (sargramostin) Reducing the duration of neutropenia caused by chemotherapy, and thereby treating and preventing infections in patients exposed to chemotherapy; inhibition of growth or eradication of Mycobacterium intracellular avium (MAC) infection Prostaglandins High blood pressure, hypertension Cyclosporin Transplant rejection Vasopressin Bed-wetting; antidiuretic Cromoline Sodium Asthma; allergies Vancomycin Treatment and prevention of infections caused by microbes resistant to antimicrobial therapy, including without limitation, resistant to metaphillin Staphalococcus aureus and Staph. epidermiditis

Active substances Disease and physiological effect Gallium nitrate Osteoporosis; Paget's disease; suppression of osteoclasts; the promotion of osteoblastic activity, hypercalcemia, including cancer associated with hypercalcemia, urethral (in the urinary tract) malignant formations; exhibits antitumor activity; cancers, including urethral cancers and bladder cancers; swelling of the lymphatic tissue; malignant tumors (including bladder cancer); leukemia; therapy of bone metastases (and associated pain); multiple myeloma, weakening of the immune response, including rejection of allogeneic transformation; the destruction of iron metabolism; activation of cell migration; wound healing; mitigation or treatment of infectious processes caused by mycobacterium species, including, without limitation, mycobacterium tubercolosis, and mycobacterium avium complex Desferrioxamine (DFO) Iron overload Parathyroid hormone (PTH), including its fragments Osteoporosis; bone disease

Active substances Disease and physiological effect Antimicrobial agents Infections, including, without limitation, infections caused by gram-positive bacteria Vitamins Treatment and prevention of vitamin deficiency

Active substances Disease and physiological effect Bisphosphonates, including alendronate, chlodronate, etidronate, ibandronate, incadronate, mindronate, neridronate, olpadronate, pamidronate, risedronate, tiludronate, zoledronate, EB1053, and YH529 Osteoporosis; Paget's disease; tumors and metastases in the bones (and associated pain); breast cancer including as an immunostimulating therapy for early stage breast cancer; therapy of bone metastases (and associated pain), including bone metastases associated with breast cancer, prostate cancer and lung cancer; suppression of osteoclast; the promotion of osteoblastic activity; treatment and / or prevention of loss of bone mineral density; multiple myeloma; prevention of bone complications associated with malignant osteolysis; fibrotic dysplasia; pediatric idiopathic osteopsatirosis; hypercalcemia, urethral (in the urinary tract) malignant tumors; sympathetic reflex dystrophy syndrome, acute back pain after a crushed fracture of the spine, chronic inflammatory articular infiltrate, renal bone disease, extracostal calcification, intoxication with analgesic, vitamin D, periarticular ossification

Active substances Disease and physiological effect Anti-migraine drugs such as BIBM-4096BS BIBN4096BS - (1-piperidinecarboxamide N- [2- [5-amino-1 - [[4- (4-pyridinyl) -1-piperazinyl) carbonyl] pentyl] amino] -1- [ (3,5-dibromo-4-hydroxyphenyl) methyl] -2-oxoethyl] -4 (1,4-dihydro-2-oxo-3 (2H0-quinazolinyl) - [R- (R *, S *)] - ) and other protein antagonists genetically related to calcitonin, sumatriptan succinate; Anti-migraine drugs; genetically related calcitonin peptide antagonist Glucagon Improving glycemic control (for example, treating hypoglycemia and controlling glycemic reactions), obesity; diagnostic tool for radiological examination of the stomach, duodenum, small intestine and colon; treatment of acute cardiovascular poisoning, including but not limited to calcium channel blockers, beta blockers GLP-1, exidin-3, exidin-4 Diabetes improved glycemic control (for example, treatment of hypoglycemia and control of glycemic reactions), obesity

Active substances Disease and physiological effect Dipeptidyl Peptidase IV Inhibitors (DPP-4) Diabetes improved glycemic control (for example, treatment of hypoglycemia), obesity Vaccines, including vaccines against anthrax or plague, flu and herpes; Prevention and course of a milder form of disease and infection YY peptide (PYY) and PYY-like peptides Obesity, Diabetes, Food Disorders, Insulin Immunity Syndromes

[244] For example, one embodiment of the present invention is a method for treating patients with or predisposed to diabetes by administering insulin in a pharmaceutical composition of the invention. Other active substances, including those listed in the table above, can also be used in conjunction with the pharmaceutical compositions of the present invention.

[245] Upon subsequent administration, the active substance present in the composition or in a unit dosage form is drawn into the blood circulation. The bioavailability of a substance can be easily assessed by measuring known pharmacological activity in the blood, for example, an increase in blood coagulation time caused by heparin, or a decrease in calcium circulation levels caused by calcitonin. Periodically, the circulation levels of the active substance itself can be directly measured.

Additives

[246] The solid pharmacological composition and unit dosage form of the present invention may contain other active substances and pharmaceutically acceptable additives, for example, fillers, carriers, diluents, stabilizers, plasticizers, binders, glidants, disintegrants, disintegrants, lubricants, dyes , film-forming substances, flavors, flavors, sugars, sugar substitutes, preservatives, metered fillers, surfactants and any combination of any of the above x substances. Preferably, these additives are suitable for pharmaceutical use, such as those described in the monograph Remington's, The Science and Practice of Pharmacy, (Gennaro, AR, ed., 20 ^th edition, 2003, Mack Pub. Co.), issued here reference.

[247] Suitable binders include, but are not limited to, starch, gelatin, sugars (such as sucrose, molasses, and lactose), calcium hydrogen phosphate dihydrate, natural and synthetic gums (such as gum arabic), sodium alginate, carboxymethyl cellulose, methyl cellulose, polyvinylpyrrolidone, polyethylene glycol, ethyl cellulose and wax.

[248] Suitable glidants include, but are not limited to, talc and silicon dioxide (quartz) (eg, fused silica and colloidal silicon dioxide).

[249] Suitable disintegrants include, but are not limited to, starches, sodium starch glycolate, croscarmellose sodium, crospovidone, clays, cellulose (eg, purified cellulose, methyl cellulose and sodium carboxymethyl cellulose), alginates, pregelatinized corn starch, such as, and carob, karai, pectin and tragacanth resins). A preferred disintegrant is sodium starch glycolate.

[250] Suitable excipients include, but are not limited to, starches (such as rice starch), microcrystalline cellulose, lactose (eg, lactose monohydrate), sucrose, dextrose, mannitol, calcium sulfate, calcium sulfate and tricalcium sulfate.

[251] 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 oils.

[252] Suitable surfactants include, but are not limited to, sodium lauryl sulfate, hydroxylated soya lecithin, polysorbates, and block copolymers of propylene oxide and ethylene oxide.

Dosage forms

[253] The solid pharmaceutical composition of the present invention, which contains the active substance and the carrier reagent, can be defined as a solid unit dosage form. The dosage form may, for example, be in the form of a tablet, sachet or capsule, in particular a hard or soft gelatin capsule. The dosage form can provide instant, prolonged or controlled release of the carrier reagent, heparin and optionally additional active substances.

[254] A solid pharmaceutical composition and a solid unit dosage form can be prepared as follows. The carrier reagent in solid form is processed (for example, crushed through a 35 mesh sieve (number of sieve openings per linear inch)) to obtain a powder having relatively small and preferably identical particle sizes. The carrier reagent is then mixed with the active substance and, optionally, a filler and / or moisturizer using, for example, a V-mixer or a similar apparatus for producing a powder mixture.

[255] A moisturizer mixture is separately prepared by mixing the moisturizer, heparin and carrier reagent. The mixture may also contain, for example, water. The composition of the moisturizing mixture is selected so as to wet the heparin when mixed with the above powder mixture. According to one preferred example, the moisturizing mixture is formulated so as to partially dissolve the carrier reagent when it is mixed with the powder mixture.

[256] The powder mixture is added in small quantities to the moisturizing mixture with constant stirring. To obtain a uniform composition, stirring is continued for a sufficient time (for example, 15 minutes) after the entire powder mixture has been added. The final composition is usually semi-solid, gel or liquid.

[257] The composition can then be transferred into a dosage form, such as a capsule, by methods known in the art. According to one preferred example, the final composition is packaged in soft or hard gelatin capsules (for example, Licap Capsugel size 0 hard gelatin capsules). Other suitable methods are described in US Pat. Nos. 6,605,298, 6,458,383, 6,261,601, 5,714,477, and 3,510,561; US Patent Publication Nos. 2003/0077303 and 2001/0024658; and International Publication No. WO 88/10117, which are all designed here by reference.

EXAMPLES

[258] The following examples demonstrate the present invention without limitation. All processes are by weight unless otherwise indicated.

[259] The compounds listed below were investigated by proton nuclear magnetic resonance (1 H NMR) methods on a Bruker® spectrometer at 300 MHz (Bruker-Physik AG, Silberstreifen, GERMANY) or on a JEOL spectrometer at 400 MHz (JEOL USA, Inc., Peabody, MA) using dimethyl sulfoxide (DMSO-d6) as a solvent, unless otherwise indicated.

[260] Analysis was performed on a liquid chromatograph / mass spectrometer (LC-MS) using Agilent Technologies (Palo Alto, California), LC / MS 1100 (single quadruplet) having the following parameters:

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

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

[263] Gradient elution: 4 minute linear gradient 0-100% B; total time for one input is 11 minutes.

[264] Injection sample volume: 5 μl.

[265] Column: ZORBAX Rapid Resolution Cartridge, SB-C18, 2.1 × 30 mm, 3.5 μm.

[267] Particle size, catalog # 873700-902.

[268] Column temperature: 40 ° C.

[269] UV detection at 244 nm.

[270] MSD parameters:

[271] Source: API-ES, positive polarity.

[272] Scan options:

Mass Range: 125.00-600.00

Fragmenter: 60 V

Growth: 1.0 EMB

Threshold: 150

Spray chamber:

Gas temperature 350 gr.

Drying gas: 12.0 l / min

Pressure; 275.79 kPa

Permissible load 4000 V

positive / negative.

Example 1 - Preparation of Compounds 1-22

[273] Compounds 1-22 were obtained according to the method described in US patent No. 6,384,278, which is framed here by full reference.

[274] The corresponding N-substituted aniline is mixed with the corresponding dicarboxylic acid monoester and heated in xylene in the presence of boric acid as a catalyst.

Example 2 - Preparation of Compounds 23-34 and 59

[275] A dried, 200 ml round bottom flask with 3 necks was equipped with a Teflon-coated magnetic stirrer and a Dean-Stark trap with a vacuum jacket, on top of which a reflux condenser was installed with an inlet adapted to supply nitrogen. N-isopropyl-N-phenylamine (8.11 g, 60 mmol), boric acid (0.93 g, 15 mmol) and xylene (88 ml) were placed in a reaction vessel. To the reaction mixture, 7-ethoxy-7-oxoheptanoic acid (11.29 g, 60 mmol) was added in one portion with stirring. To return the phlegm, the reaction mixture was heated using a heating mantle. Water that began to be released during azeotropic distillation was trapped in a Dean-Stark trap. After 16 hours of irrigation, water was collected and the reaction mixture was allowed to cool to room temperature. The reaction mixture was diluted with ethyl acetate (100 ml), and washed with 2 N. aqueous HCI solution (50 ml), and then saturated sodium bicarbonate solution (60 ml). Most of the organic solvent was removed in vacuo. To the residue was added 2 N. 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 the aqueous phase was carefully separated from the organic layer, it was evaporated to remove any residual ethyl acetate. Ice was added to the aqueous solution, followed by an aqueous HCl solution (2N, 60 ml), which resulted in the precipitation of a white solid. Before collecting the precipitate using a sintered funnel, stirring was continued for an additional 30 minutes. The collected white solid was washed effectively with water and hexane before being vacuum placed at room temperature for 12 hours to obtain 7.49 g (45%) of 7- [isopropyl (phenyl) amino] -7-oxoheptanoic acid as a white solid . HPLC: single peak at 4.83 min .; Mr: 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). Weight (M + 1): 278. Anal. Calculation for C ₁₆ H ₂₃ NO ₃ : C, 69.29; H, 8.36; N 5.05. Found: C, 69.06; H, 8.45; N, 4.99.

[276] Compounds 24-34 and 59 were prepared from the corresponding starting materials using the above method.

Compound (24)

[277] HPLC: single peak at 4.43 min. Mass (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)

[278] HPLC: single peak at 4.62 minutes Mass (M + 1): 264. ¹ H NMR (400 MHz, 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)

[279] HPLC: single peak at 4.19 min. Mass (M + 1): 250. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ: 0.65 (d, 3H), 0.84-0.86 (1, 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)

[280] HPLC: single peak at 3.92 min. Mass (M + 1): 250. ¹ H NMR (400 MHz, 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)

[281] HPLC: single peak at 3.72 min. Mass (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)

[282] HPLC: single peak at 3.88 min. Mass (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 (100 MHz, DMSO-d ₆ ) δ: 28.81, 29.0, 36.5, 129.32, 129.58, 132.0, 142.66, 170.58, 173.63.

Compound (30)

[282] HPLC: single peak at 4.82 min. Mass (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)

[283] HPLC: single peak at 4.44 min. Mass (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)

[284] HPLC: single peak at 4.81 min. Mass (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)

[285] HPLC: single peak at 5.48 min. Mass (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., 1H).

Compound (34)

[286] HPLC: single peak at 4.52 min. Mass (M + 1): 282. ¹ H NMR (400 MHz, DMSO-s1b) 5: 1.09 (t, 4H), 1.39 (t, 4H), 1.93 (t, 2H), 2.10-2.14 (t, 2H ), 3.09 (s. ZN), 3.75 (8 ZN), 7.19 (t, 2H), 7.30 (t, 2H), 11.91 (br., 1H).

Compound (59)

[287] HPLC: single peak at 4.71 minutes Mass (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 (8, 1H).

Example 3 - Preparation of Compounds 111-139

Compound 111 - 4-Oxo-4-phenylbutyric acid:

[289] 10 g (56 mmol) of 3-benzoylpropionic acid (commercially available from Sigma-Aldrich Co., St. Louis, Mo.) was added to 10 ml of water. The mixture was stirred and 28 ml of 2N was added. sodium hydroxide (aqueous). The final solution was stirred for 2 hours and, after freeze-drying, a solid product was collected. 1 H NMR (d6-DMSO): δ 7.9, d, 2H, (aryl H); δ 7.6, t, 1H, (aryl H); δ 7.5, t, 2H, (aryl H); δ 3.1, t, 2H (CH ₂ α to carbonyl); δ 2.2, t, 2H (CH ₂ α to COOH); COOH peak was not observed due to the presence of water in the sample.

Compound 113-10- (4-Hydroxy-phenyl) -10-oxodecanoic acid:

[290] Decanedic acid (20 g, 296 mmol) and acetic anhydride (280 ml, 2.96 mol) were placed in a 500-ml flask equipped with a reflux condenser in an inert atmosphere. For irrigation, the mixture was heated for 5 hours. Acetic acid and excess acetic anhydride were removed under reduced pressure. The product was used without further purification.

[292] To a 500 ml flask equipped with a mechanical stirrer, oxacycloundecan-2,11-dione (20 g, 108.5 mmol), phenol (10.22 g, 108.5 mmol) and 200 ml of disulfide were added in an inert atmosphere carbon. Aluminum (III) trichloride (72.34 g, 542 mmol) was added and the reaction mixture was stirred for 72 hours. Carbon disulfide was decanted and ice was carefully added until most of the mixture had dissolved. Insoluble matter was collected by vacuum filter and washed with 2 × 100 ml of water. Then, the solid was dissolved in 100 ml of 1 M aqueous sodium hydroxide and carefully acidified with 1 M hydrochloric acid (aqueous) to pH = 7.5. The resulting dry residue was removed by filtration and the initial solution was continued to acidify to pH 2.5. The crude product precipitate was filtered off and washed with 1 × 100 ml of water. The crude product was dissolved in 100 ml of a 1 M aqueous solution of sodium hydroxide, carefully acidified with a 1 M aqueous solution of hydrochloric acid to pH = 7.5, and precipitated impurities were filtered off. The initial solution was then acidified to pH 2. The crude product was filtered off and washed with 2 × 50 ml of water. Then the product was recrystallized from acetone. The isolated product (1.2 g, 4%) was collected by filtration. Found: C 69.00, H 7.81%; C ₁₆ H ₂₂ O ₄ requires C: 69.04, H: 7.97% 1H NMR (d6-DMSO): δ 12.0, bs, 1H (COOH); δ 10.3, bs, 1H (aryl-hydroxyl); δ 7.8 d, 2H (aryl H); δ 6.8, d, 2H, (aryl H); δ 2.9, t, 2H (CH ₂ α to carbonyl); δ 2.2, t, 2H (CH ₂ α to COOH); δ 1.5, multiplet, 4H (CH ₂ groups β to carbonyl and β to COOH), δ 1.3, multiplet, 8H (other CH ₂ groups).

Compound 114 - 10- (2-Hydroxy-phenyl) -10-oxodecanoic acid:

[293] Methylene chloride (50 ml), 9-bromononanol (7.63 g, 34.2 mmol) and trimethylsilyl chloride (4.5 ml, 35.5 mmol) were placed in a 100 ml flask and stirred under nitrogen for 20 minutes. Then triethylamine (5.0 ml, 35.9 mmol) was added and the final reaction mixture was stirred for 2 hours at room temperature. Then the reaction mixture was diluted with 80 ml of hexane, filtered, and then concentrated under reduced pressure. The final residue was diluted again with 80 ml of hexane, filtered, and then concentrated under reduced pressure to obtain 9.7 g (96%) of a yellow liquid, which was used without further purification.

[294] 5.69 g (19.3 mmol) of (9-bromo-nonyloxy) trimethylsilane was added dropwise in an inert atmosphere to a 50 ml flask containing magnesium metal (0.59 g, 24.3 mmol), 20 ml of tetrahydrofuran and used a small crystal of iodine to initiate the Grignard reaction. In a 100 ml flask in an inert atmosphere, a solution of salicylic aldehyde (2.1 ml, 19.7 mmol) in 20 ml of tetrahydrofuran was cooled using an external ice bath. Then, the cooled aldehyde solution was treated with 1.0 M lithium bis (trimethylsilyl) amide (20.0 ml, 20 mmol). After stirring for 1 hour, the Grignard reaction mixture was cooled using an external ice bath. Then, with constant stirring, the cooled Grignard reaction mixture was added dropwise through the cannula to the aldehyde solution over a 5 minute period. The final reaction mixture was allowed to warm to room temperature and continued to stir overnight. 40 ml of ethyl acetate was poured into the reaction mixture and quenched with 15 ml of a saturated aqueous sodium bicarbonate solution. The organic layer was separated and washed with 2 × 25 ml portions of a 4% aqueous hydrochloric acid solution, and then 1 × 20 ml portions of brine. The organic layer was dried over sodium sulfate, filtered, and the solvent was removed under reduced pressure. Residual salicylaldehyde was removed by Kugelroch distillation and the final precipitate was used without further purification.

[295] 1- (2-hydroxy-phenyl) -undecan-1,11-diol (5.0 g, 18.9 mmol) and 50 ml of dimethylformamide were placed in a 100 ml flask. Pyridine dichromate (32.9 g, 87.5 mmol) was added thereto. (The additive was moderately exothermic.) The reaction mixture was stirred at room temperature overnight. The reaction mixture was poured into 50 ml of ethyl acetate and washed with 200 ml of water, 30 ml of a 4% aqueous hydrochloric acid solution, 30 ml of water, and finally 30 ml of brine. Then the organic layer was stirred for 15 minutes with 10 g of silica gel, dried with sodium sulfate, filtered, and the solvent was removed under reduced pressure. The crude off-white product was recrystallized from ethanol / water. The product (0.5 g, 10%) was isolated as an off-white solid with a melting point of 85-87 ° C. Organic elemental analysis: Found: C 69.01, H 8.36%; C ₁₆ H ₂₂ O ₄ requires C: 69.54, H: 8.02% 1H NMR (d6-DMSO): δ 12.0, s, 1H (COOH); δ 7.9 dd, 1H (aryl H); δ 7.5, dt, 1H, (aryl H); δ 6.9, complex multiplet, 2H (aryl H), 3.1, t, 2H (CH ₂ α to carbonyl); δ 2.2, t, 2H (CH ₂ α to COOH); δ 1.6, multiplet, 2H (CH ₂ β to carbonyl), δ 1.5, multiplet, 2H (CH ₂ β to COOH), δ 1.3, multiplet, 8H (residue of the CH ₂ group).

Compound 115 - 4- (4-Methoxy-phenyl) -4-oxo-butyric acid:

[296] In a 500 ml round bottom flask equipped with a magnetic stirrer, 5.25 ml (48.3 mmol) of anisole, 4.83 g (48.3 mmol) of succinic anhydride, 125 ml 1 were placed in an inert atmosphere (nitrogen gas) , 1,2,2-tetrachlopentane and 125 ml of nitrobenzene. The reaction vessel was cooled using an external water bath and the reaction mixture was stirred for 30 minutes. Aluminum trichloride (14.2 g, 106.4 mmol) was added to a pale yellow solution, which was then stained dark reddish brown. The ice bath was removed and the reaction mixture was continued to stir at room temperature for 36 hours. Then the reaction mixture was again cooled with an external water bath. Prepared acidic solution by pouring a solution of 1 N. hydrogen chloride in a 100 ml beaker filled with ice. This solution was carefully added to the reaction mixture, initially dropwise until the reaction mixture became clear with the appearance of a white precipitate. After this point, a 10 ml portion was carefully added to the reactivity sample, and then the remaining ice / acid mixture was added. A second 100 ml ice / acid mixture was added, the outer ice bath was removed, and the clarified emulsion was stirred for 2 hours. The white precipitate was recovered from the emulsion by a vacuum filtration method. This solid was dissolved in 300 ml of a 0.3 M sodium hydroxide solution, washed with 100 ml of ethyl acetate, and acidified to ~ pH 11 M with hydrochloric acid. The white precipitate collected by the vacuum filtration method was washed with 3 × 100 ml deionized water and dried. The product (4.7 g, 47%) was isolated as a white solid with a melting point of 149-150 ° C. Organic elemental analysis: Found: C 63.52, H 5.78%; C ₁₁ H ₁₂ O ₄ requires C: 63.45, H: 5.81% 1H NMR (d6-DMSO): δ 12.2, s, 1H (COOH); δ 7.9 d, 2H (aryl H); δ 7.0, d, 2H, (aryl H); δ 3.8, s, 3H (OMe H's); δ 3.2, t, 2H (CH ₂ α to carbonyl); δ 2.5, t, 2H (CH ₂ α to COOH).

Compound 116 - 4-Methoxy-phenyl) -5-oxopentanoic acid:

[297] Compound 116 was obtained similarly to compound 15, except for the use of glutamic anhydride instead of succinic anhydride, melting point 141-142 ° C. Found: C 64.65, H 6.34%; C ₁₂ H ₁₄ O ₄ requires C: 64.85; H: 6.35% 1H NMR (d6-DMSO): δ 12.2, s, 1H (COOH); δ 7.9 d, 2H (aryl H); δ 7.0, d, 2H, (aryl H); δ 3.8, s, 3H (OMe H's); δ 3.0, t, 2H (CH ₂ α to carbonyl); δ 2.3, t, 2H (CH ₂ α to COOH)); δ 1.8 quintuplet, 2H (CH ₂ between the other two).

[298] Compound 117 was purchased from Aldrich (St. Louis, MO), catalog number 514683.

[299] Compound 118 was purchased from Aldrich (St. Louis, Mo.), catalog number B12687.

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

[301] Compound 120 was purchased from Reike, Aldrich (St. Louis, Mo.), catalog number 7013D.

[302] Compound 121 was purchased from Reike, Aldrich (St. Louis, Mo.), catalog number 7148C and

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

[303] 5- (4-Isopropyl-phenyl) -5-oxo-pentanoic acid (5 g, 21.3 mmol) was dissolved in 75 ml of ethanol in a 250 ml flask. Sodium hydroxide (0.85 g, 21.3 mmol) was added and the reaction mixture on a rotary evaporator was stirred overnight under reduced pressure. The solid was dried in vacuo and used without further purification. Found: C 60.24, H 6.66, Na 9.21%; C ₁₄ H ₁₇ O ₃ Na requires C: 61.28, H: 6.98, Na 8.38% 1H NMR (D2O): 5 7.7, d, 2H (aryl H); δ 7.2 d, 2H (aryl H); δ 2.9, t, 2H (CH ₂ α to carbonyl); δ 2.8, multiplet, 1H, (CH of isopropyl group); δ 2.1, t, 2H (CH ₂ α to COOH); δ 1.8, q, 2H (CH ₂ β for both carbonyl and COOH), δ 1.1, d, 6H (CH ₃ 's of the isopropyl group).

[304] Compound 122 was purchased from Aldrich (St. Louis, Mo.), catalog number B13802.

[305] Compound 123 was purchased from Reike, Aldrich (St. Louis, MO), catalog number 7060B.

[306] Compound 124 was purchased from Fischer-Scientific (Hampton, New Hampshire), Acros, catalog number 17.522.62

[300] Compound 125 was purchased from Reike, Aldrich (St. Louis, Mo.), catalog number 7011D.

[301] Compound 126 was purchased from Reike, Aldrich (St. Louis, MO), catalog number 7036B.

[302] Compound 128 was purchased from Reike, Aldrich (St. Louis, MO), catalog number 7012D.

[303] Compound 129 was purchased from Reike, Aldrich (St. Louis, Mo.), catalog number 7012B.

[304] Compound 130 was purchased from Reike, Aldrich (St. Louis, MO), catalog number 7055B

[305] Compound 132 was purchased from Reike, Aldrich (St. Louis, MO), catalog number 7005b.

Compound 133 was purchased from Reike, Aldrich (St. Louis, MO), catalog number 7036F.

[307] Compound 134 was purchased from Reike, Aldrich (St. Louis, Mo.), catalog number 7144D

[308] Compound 136 was purchased from Reike, Aldrich (St. Louis, Mo.), catalog number 7144B.

[309] Compound 138 was purchased from Reike, Aldrich (St. Louis, MO), catalog number 7036D.

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

[310] Decanedic acid (20 g, 296 mmol) and acetic anhydride (280 ml, 2.96 mol) were placed in a 500 ml flask equipped with a reflux condenser in an inert atmosphere. For irrigation, the mixture was heated for 5 hours. Acetic acid and excess acetic anhydride were removed under reduced pressure. The product was used without further purification.

[311] A pre-synthesized oxacycloundecan-2,11-dione (37.95 g, 206 mmol), 1,4-diacetoxybenzene (20 g, 103 mmol) and 200 were placed in a 500-ml flask equipped with a mechanical stirrer in an inert atmosphere. ml of carbon disulfide. Aluminum (III) trichloride (68.7 g, 515 mmol) was added and the reaction mixture was stirred for 72 hours. Carbon disulfide was decanted and ice was carefully added until most of the mixture had dissolved. The insoluble matter was collected by vacuum filtration and washed with 2 × 100 ml of water. Then, the solid was dissolved in 50 ml of a 1 M aqueous sodium hydroxide solution and stirred for 1 hour. The solution was acidified with 1 M aqueous hydrochloric acid to pH = 2. The crude product precipitate was collected by filtration, redissolved in acetonitrile (50 ml) and methylene chloride (15 ml) and allowed to slowly precipitate for a week. The resulting brown product was filtered off and recrystallized from a mixture of acetic acid: water 10: 3. The product was isolated (0.8 g, 3%) by filtration. Found: C 65.55, H 7.69%; C ₁₆ H ₂₂ O ₅ requires C: 65.29, H: 7.53% 1H NMR (d6-DMSO): δ 12.0, s, 1H (COOH); δ 11.4, s, 1H (aryl-hydroxyl); δ 9.2, s, 1H (aryl-hydroxyl); δ 7.2 d, 1H (aryl H); δ 7.0, dd, 1H, (aryl H); δ 6.8, d, 1H (aryl H), 3.0, t, 2H (CH ₂ α to carbonyl); δ 2.2, t, 2H (CH ₂ α to COOH); δ 1.6, multiplet, 2H (CH ₂ β to carbonyl), δ 1.5, multiplet, 2H (CH ₂ β to COOH), δ 1.3, multiplet, 8H (residue of the CH ₂ group).

Example 4 - Preparation of Compounds 140-151

[312] In General, these compounds were prepared in four stages. First, salicylic acid, suitably substituted, and 3-aminobutyric acid ethyl ester with ethylene dichloride (EDC) / hydroxybenzene triazole (GBT) / dichloromethane (DCM) were mixed. Second, A-15 / A-21 base ion-exchange resin (commercially available in Rohm and Haas, Philadelphia, PA) was added. Third, after partial processing, the product was reacted with potassium trimethylsilanolate (TMSC) / tetrahydrofuran (THF). Fourth, IRC-50 resin (Rhohm & Haas, Philadelphia, PA) was added.

[313] Salicylic acid (4.57 mmol), DCM (10 ml), EDC (1.05 g, 5.48 mmol), GBT (838 mg, 5.48 mmol), DMF (2) were placed in each scintillation tube ml) and ethyl 3-aminobutyrate (600 mg, 4.57 mmol). All tubes were tightly sealed with stoppers, placed on a J-Kem reaction block (J-Kem Scientific Inc., St. Louis, Mo.), and shaken with heating (150 rpm, 35 ° C.) overnight. Controlled by TLC, all reactions have one dominant point. Amberlyst-21 and Amberlyst-15 resins (approximately 2.5 g, 11 mmol) were added to each vial and continued to shake at ambient temperature overnight. The reaction mixture was filtered, the resins washed with DCM (2 × 5 ml), and the combined filtrates of each reaction mixture were collected in a clean scintillation tube. The filtrates were purged with a stream of nitrogen to a volume of approximately 2 ml.

[314] A 1.2 M solution of potassium trimethylsilonolate (TMSC) in THF (10 ml, 12 mmol) was placed in each tube. More THF was added to some reaction mixtures when it was necessary to prepare suspensions suitable for shaking. All tubes were tightly closed with stoppers, placed on a J-Kem reaction block, and shaken with heating (150 rpm, 60 ° C for 6 h). The reaction block was cooled and IRC-50 resin (3 g, 30 mmol) was added to each tube to quench the potassium salt. If necessary, DCM was added to suspend the resin and facilitate shaking. The reaction mixture shook all night. The reaction mixtures were filtered, the resins were washed with DCM (2 × 5 ml), if necessary, to dissolve the solids, washed with DMF, and the combined filtrates of each reaction mixture were collected in a clean scintillation tube. At this stage, small aliquots of the filtrate were taken and diluted with 1: 1 ACN / H ₂ O mixture for LC-NM. The filtrates were purged with a stream of nitrogen. To remove traces of DMF, the tubes were placed in a vacuum oven at 50 ° C.

[315] According to LC-MS analysis, some reaction mixtures still contained significant amounts of ether. These mixtures were re-treated with TMSC. A 1.2 M solution of TMSC in THF (8 ml, 9.6 mmol) was added to each tube. All tubes were tightly closed with a lid, placed in a Pierce reaction block, stirred and heated (60 ° C, 5 hours). The reaction block was cooled, and IRC-50 resin (2 g, 20 mmol) was added to each tube to quench the sodium salt. If necessary, DCM was added to suspend the resin and facilitate shaking. The reaction mixture was stirred all weekend. The reaction mixtures were filtered through a layer of quartz, and the quartz was washed with DCM (2 × 5 ml), then with a 2: 5 MeOH / DCM mixture (3 × 7 ml), and the combined filtrates of each reaction mixture were collected in a clean scintillation tube. At this stage, small aliquots of the filtrate were taken and diluted with 1: 1 ACN / H ₂ O for LC-MS. The filtrates were purged with a stream of nitrogen.

[316] All other reaction mixtures, the rest from the first TMSC treatment, were placed in a 10: 1 DCM / MeOH mixture and filtered through a quartz layer, eluting with a large amount of 10: 1 DCM / MeOH. At this stage, small aliquots of the filtrate were taken and diluted with 1: 1 ACN / H ₂ O for LC-NM. The filtrates were purged with a stream of nitrogen.

Alternative Preparation of Compound 140-151:

[317] 3,5-diisopropylsalicylic acid (25.0 g, 112.5 mmol), GBT (20.6 g, 135.0 mmol), ethyl 3-aminobutyrate (18) were placed in a 1-liter round-bottom flask. 0 g, 123.7 mmol) and dioxane (400 ml). The resulting mixture was stirred at ambient temperature. EDC (25.9 g, 135.0 mmol) was added in portions and continued to mix overnight. HPLC analysis of the reaction mixture at this stage showed the presence of GBT, possibly trace amounts of the initial salicylic acid, and one new product in predominant quantities. An additional portion of EDC (5 g, 26.0 mmol) was added and allowed to mix overnight. Another HPLC analysis showed no significant changes. The reaction mixture was quenched with water (400 ml) and, using a rotary evaporator, dioxane was distilled off. The resulting oil / water mixture was poured into a 1 L separatory funnel and DCM (400 ml) was added. A large amount of white solid formed. Trying to separate the layers, EtOAc was added, but to no avail. The mixture was poured from the separation column and light organic fractions were distilled off from it on a rotary evaporator. The water / oil mixture was extracted with EtOAc (500 ml, then 200 ml). The combined EtOAc layers were washed with an aqueous HCl solution (10%, 2 × 200 ml), an aqueous NaOH solution (10%, 2 × 200 ml) and brine (50 ml, then 200 ml). The organics were dried over Na ₂ SO ₄ and the light fractions were distilled off on a rotary evaporator to give a brown oil that contained small amounts of a white solid. HPLC analysis showed that the white solid was the residual amount of HBT, and the desired product was brown oil. Brown oil was taken with a pipette from the flask, trying to prevent solid white matter from falling into the pipette. Brown oil was placed in EtOAc (500 ml), washed with NaOH (10%, 2 × 200 ml) and dried over Na ₂ SO ₄ . EtOAc was distilled off on a rotary evaporator to give a brown oil. At this stage, the results of HPLC analysis showed one predominant peak and the absence of GBT.

[318] A viscous oil was dissolved in THF (200 ml) and TMSC (31.7 g, 247.4 mmol) was added. The resulting viscous mixture was stirred overnight. HPLC analysis indicated the end of the reaction by the presence of one peak. IRC-50 resin (37 g, 370 mmol, 1.5 eq.) And 100 ml of DCM were added to suspend the resin, then stirred for several hours. Filtered, washed with DCM resin (3 × 50 ml) and concentrated on a rotary evaporator to a brown oil. An attempt to recrystallize from an ACN / acetone mixture was unsuccessful. Based on solubility, it was determined at this stage that the potassium salt was the predominant substance. The oil was placed in a mixture of H ₂ O / ACN, heated until it became transparent, filtered off in a hot state, and cooled to room temperature. The filtrate was treated with an aqueous HCl solution, the resulting solid precipitate was isolated and crushed into powder to obtain E1528: 9.13 grams, HPLC: retention time 6.7 min 100%, KF 0.47, NMR consistent with the structure, elementary analysis theoretically C: 66.11, H: 8.21 , N: 4.54, found C: 65.62, H: 8.19, M: 4.46.

Connection number and name MW MS (M + H) Will approximate. purity percentage based on LC-MS Compound 140
3- (2-hydroxy-benzoylamino) butyric acid 223.2306 224 83 Compound 141 381.0326 382 71 3- (3,5-dibromo-2-hydroxy-benzoylamino) butyric acid Compound 142 292.1206 292 77 3- (3,5-dichloro-2-hydroxy-benzoylamino) butyric acid Compound 143 475.0234 476 82 3- (2-hydroxy-3,5-diiodo-benzoylamino) butyric acid Compound 144 237.2577 238 75 3- (2-hydroxy-3-methyl-benzoylamino) butyric acid Compound 145 257.6756 258 82 3- (4-chloro-2-hydroxy-benzoylamino) butyric acid Compound 146 253.2571 254 75 3- (2-hydroxy-4-methoxy-benzoylamino) butyric acid Compound 147 302.1316 303 82 3- (5-bromo-2-hydroxy-

benzoylamino) butyric acid Compound 148 257.6756 258 78 3- (5-chloro-2-hydroxy-benzoylamino) butyric acid Compound 149 253.2571 254 77 3- (2-hydroxy-5-methoxy-benzoylamino) butyric acid Compound 150 237.2577 238 82 3- (2-hydroxy-5-methyl-benzoylamino) butyric acid Compound 151 307.3931 308 89 3- (2-hydroxy-3,5-diisopropyl-benzoylamino) butyric acid

Example 5 - Preparation of Compounds 152-160

[319] Compound 152 — was purchased from Transworld Chemical (South Melbourne, Australia).

[320] Compound 153 — was purchased from Lancaster (Windham, NH).

[321] Compound 154 - was purchased from Avocado (Heysham, Lancashire, England).

[322] Compound 155 — was purchased from Aldrich, catalog number 42919 (St. Louis, MO).

[323] Compound 156 — was purchased from Sigma-Aldrich (St. Louis, MO).

[324] Compound 157 — was purchased from Sigma (St. Louis, MO).

[325] Compound 158 — was purchased from Matrix Scientific (Columbia, SC).

Compound HPLC
Time
retention HPLC
Proto
count KF value Temperature
Ra
melting
interval
values CHNC C CHNC H Chnr
FROM 152 5.41 0 153 5.1 min 0.1% TFU 0 69.76 5.46 154 6.2 min 0.1% TFU 0 74.98 6.29 155 5.21 0 156 5.82 min 0.1% TFU 0 157 5.42 0 184-186 73.67 5.3 72.56 158 5.47 0 110-112 74.36 5.82 74.39 159 5.56 min 63.47 5.39 62.65 160 5.30 0.3 67-70 73.45 5.32 73.08

Compound 160:

[326] Potassium hydroxide (10.37 g, 184.8 mmol) was ground in a mortar and pestle to a powder and placed in a 250-ml flask containing 75 ml of dimethyl sulfoxide and 2-hydroxy-benzoic acid methyl ester (7.03 g, 46 , 2 mmol). Benzyl bromide (7.91 g, 46.2 mmol) was added to this mixture and the mixture was prepared for 4 hours with stirring. Water (100 ml) was added and the reaction mixture was stirred for another 30 minutes. Then the reaction mixture was cooled using an external ice bath to 0 ° C and acidified with concentrated hydrochloric acid to pH 1. The mixture was extracted with 3 × 230 ml of 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 × 30 ml of water, and then 2 × 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 over vacuum for several days, and a white crystalline solid formed. The solid product was collected and then dried over vacuum. The product (8.04 g, 76%) was isolated as a white crystalline solid with a melting point of 67-70 ° C. Organic elemental analysis: Found: C 73.08, H 5.37%; C ₁₄ H ₁₂ O ₃ requires C 73.45, H 5.32%;

Example 6 - Preparation of Compounds 160-167

[326] Compound F was prepared according to the general scheme, wherein 2-hydroxybenzophenone was alkylated with bromoalkyl ether in the presence of a base, which was accompanied by decomposition of the ether portion using potassium trimethylsilanoate.

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

In a 250-ml round-bottom flask equipped with a magnetic stirrer and a reflux condenser, 10.32 g (48.2 mmol) of 2,2'-dihydroxybenzophenone and 100 ml of dimethyl sulfoxide (DMSO) were placed. Potassium hydroxide (2.91 g, 51.9 mmol), previously ground to a powder, was added to the clear solution. The reaction mixture was heated to 45 ° C. until most of the solid had dissolved. The resulting red suspension was treated with 8.80 ml (11.04 g, 49.5 mmol) of ethyl 6-bromohexanoate. After stirring for 20 hours at 25 ° C, the clear reaction mixture was diluted with aqueous 1% hydrochloric acid and methyl tert-butyl ether (MTBE). The layers were separated. The organic phase was washed with water (2 × 50 ml) and brine (1 × 40 ml), dried over sodium sulfate and concentrated. The resin was placed in 100 ml of tetrahydrofuran (THF) and treated with potassium trimethylsilanoate (15.09 g, 118 mmol). The orange solution was stirred for 20 hours at 25 ° C, diluted with 4% aqueous hydrochloric acid to pH 7.5 and washed with MTBE. The organic phase was extracted with aqueous 3% sodium bicarbonate solution. The combined aqueous phases were acidified to pH 2 with 4% aqueous hydrochloric acid and extracted with 60 ml of MTBE. This organic phase was washed with brine (1 × 40 ml), dried over sodium sulfate and concentrated. The residue was purified by flash chromatography using an 80% hexane / ethyl acetate mixture (introduced with 0.5% acetic acid). The product (4.2 g, 27%) was isolated as an off-white solid with a melting point of 89-91 ° C. Organic elemental analysis: Found: C 69.50, H 6.04%; C ₁₉ H ₂₀ O ₅ requires C: 69.50, H: 6.14% 1H NMR (d6-DMSO): δ 12.0, bs, 1H (COOH); δ 11.5, bs, 1H (OH); δ 7.5, t, 2H, (aryl H); δ 7.4, dd, 1H (aryl H); δ 7.3, dd, 1H (aryl H); δ 7.15, d, 1H (aryl H); δ 7.1, t, 1H (aryl H); δ 7.0, d, 1H (aryl H); δ 6.9, t, 1H (aryl H); δ 3.9, t, 2H, (CH ₂ α to O); δ 2.05, t, 2H (CH ₂ α to COOH); δ 1.4, m, 4H (the other two CH ₂ 's); δ 1.0, p, 2H (CH ₂ in the middle of the chain).

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

[329] Compound 161 - Sodium 8- (2- (2-hydroxybenzoyl) phenoxy) octanoate:

Using 2,2'-dihydroxybenzophenone and ethyl 8-bromoctanoate as starting materials, the title compound was prepared and then converted to the sodium salt as follows: free acid (3.56 g, 9.99 mmol) was dissolved in 40 ml of isopropanol and treated with a solution of sodium hydroxide (1.7 ml) prepared from sodium hydroxide (0.90 g, 22.5 mmol) and water (3.7 ml). Isopropanol and MTBE were added, resulting in precipitation of a solid. When this mixture was heated, most of the solid dissolved. The remaining solid was removed by filtration. The off-white solid formed upon cooling with dry ice was isolated by filtration and dried under reduced pressure. Organic elemental analysis: Found: C 65.02%, H 6.22%; C ₂₁ H ₂₃ O ₅ Na requires C 66.00, H 6.65%, ¹ H NMR (d6-DMSO): δ 12.6, bs, 1H (OH); δ 7.41, t, 1H, (aryl H); δ 7.31, t, 1H (aryl H); δ 7.27, dd, 1H (aryl H); δ 7.15, dd, 1H (aryl H); δ 7.03, d, 1H (aryl H); δ 6.97, t, 1H (aryl H); δ 6.91, d, 1H (aryl H); δ 6.65, t, 1H (aryl H); δ 3.83, t, 2H, (CH ₂ α to O); δ 1.82, t, 2H (CH ₂ α to COONa); δ 1.3, m, 4H (two more CH ₂ 's); δ 1.0, m, 6H (CH ₂ 's in the middle of the chain). ¹³ 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 .

[330] Compound 162-5- (2- (2-hydroxybenzoyl) -4-methoxyphenoxy) valerianic acid (main isomers are shown): LC-MS analysis: m + 1 peak (345). The results of 1H NMR analysis: (d6-DMSO): δ 12.4, ds, 1H (COOH); δ 11.9, bs, 1H (OH); δ 7.47, t, 1H, (aryl H); δ 7.26, dd, 1H (aryl H); δ 7.14, d, 1H (aryl H); δ 7.13, d, 1H (aryl H); δ 7.03, t, 1H (aryl H); δ 6.49, d, 1H (aryl H); δ 6.42, dd, 1H (aryl H); δ 3.95, t, 2H, (CH ₂ α to O); δ 3.79, s, 3H, (CH ₃ O); δ 2.07, t, 2H (CH ₂ β to 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, 33.05, 27.80, 20.77.

[331] Compound 163-5- (2- (2-hydroxybenzoyl) phenoxy) valerianic acid: LC-MS analysis: m + 1 peak detected (315). Results of 1 H NMR analysis: (d6-DMSO): δ 11.9, bs, 1H (COOH); δ 11.5, bs, 1H (OH); δ 7.50, dt, 1H, (aryl H); δ 7.48, dt, 1H, (aryl H); δ 7.35, dd, 1H (aryl H); δ 7.25, dd, 1H (aryl H); δ 7.14, d, 1H (aryl H); δ 7.06, t, 1H (aryl H); δ 6.96, d, 1H (aryl H); δ 6.85, t, 1H (aryl H); δ 3.93, t, 2H, (CH ₂ α to O); δ 2.06, t, 2H (CH ₂ α to COOH); δ 1.42, p, 2H (CH ₂ in the chain); δ 1.29, 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, 27.75, 20.70.

[332] Compound 164-5- (2- (2-hydroxy-5-methoxybenzoyl) -4-methoxyphenoxy) valerianic acid: LC-MS analysis: m + 1 peak detected (375). Results of 1H NMR analysis: (d6-DMSO): δ 12.4, bs, 1H (COOH); δ 12.0, bs, 1H (OH); δ 7.25, d, 1H, (aryl H); δ 7.21, d, 1H, (aryl H); δ 6.66, d, 1H (aryl H); δ 6.62, dd, 1H (aryl H); δ 6.48, d, 1H (aryl H); δ 6.42, dd, 1H (aryl H); δ 3.96, t, 2H, (CH ₂ α to 0); δ 3.81, s, 3H, (CH ₃ O); δ 3.80, s, 3H, (CH ₃ O); δ 2.08, t, 2H (CH ₂ α to COOH); δ 1.48, p, 2H (CH ₂ in chain); δ 1.34, 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, 55.69, 55.48, 33.06, 27.75, 20.77.

[333] (Compound 166) 4- (2- (2-hydroxybenzoyl) phenoxy) butyric acid: LC-MS analysis: m + 1 peak detected (333). Results of 1H NMR analysis: (d6-DMSO): δ 12.0, bs, 1H (COOH); δ 7.46, m, 2H (aryl H); δ 7.33, dt, 1H (aryl H); δ 7.29, d, 1H (aryl H); δ 6.82, t, 1H (aryl H); δ 3.77, t, 2H, (CH ₂ α to O); δ 1.85, t, 2H (CH ₂ α to COOH); δ 1.35, p, 2H (median CH ₂ in the chain). ^ 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, 29.55, 23.92.

[334] Compound 167 - 4- (2-chlorobenzoyl-4-methylphenoxy) butyric acid: LC-MS analysis: m + 1 peak detected (333). Results of 1H NMR analysis: (d6-DMSO): δ 12.4, bs, 1H (COOH); δ 12.0, bs, 1H (OH); δ 7.23, d, 1H (aryl H, o to O); δ 3.74, t, 2H, (CH ₂ α to O); δ 2.25, s, 3H, CH ₃ ); δ 1.84, t, 2H (CH ₂ α to COOH); δ 1.33, p, 2H (median CH ₂ in 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, 55.69, 55.50, 29.56, 23.85.

An example of the preparation of compounds 168-173

[335] (Compound 168) 4- (2-benzoyl-5-methoxyphenoxy) butyric acid:

4.56 g (20.0 mmol) of 2-hydroxy-4-methoxybenzophenone, 2.70 ml (3.68 g, 18.9 mmol) of ethyl 4- were placed in a 100 ml mini-block tube equipped with a magnetic stirrer. bromobutyrate and 40 ml of 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 for 20 hours at 25 ° C, the clear reaction mixture was diluted with water. The resulting solid was isolated by filtration. This solid was placed in 30 ml of tetrahydrofuran (THF) and 3.10 g (24.0 mmol) of potassium trimethylsilanoate were treated. The orange solution was stirred for 20 hours at 25 ° C, diluted with 4% aqueous hydrochloric acid to pH 7.5 and washed with MTBE. The organic phase was extracted with 3% aqueous sodium bicarbonate. The combined aqueous phases were acidified to pH = 2 with a 4% aqueous hydrochloric acid solution and extracted with 60 ml of MTBE. This organic phase was washed with brine (1 × 40 ml), dried over sodium sulfate and concentrated. The resulting solid was purified by pulverization using MTBE / hexanes mixtures. Most of the product was isolated from the mother liquor. LC-MS analysis: m + 1 established peak (315). Results of 1 H NMR analysis: (d6-DMSO): δ 12.0, bs, 1H (COOH); δ 7.6, d, 2H, (phenyl H, o to CO); δ 7.56, t, 1H (phenyl H, p to CO); δ 7.44, t, 2H (phenyl H, m to CO); δ 7.35, d, 1H (aryl H, o to CO); δ 6.64, m, 2H (aryl H, m to CO); δ 3.88, t, 2H, (CH ₂ α to O); δ 3.82, s, 3H, (CH ₃ O); δ 1.84, t, 2H (CH ₂ α to COOH); δ 1.53, p, 2H (median CH ₂ in 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.

[336] Other carrier reagents were synthesized in the same manner: Compound 169, Compound 170, Compound 171, Compound 172, and Compound 173.

[337] (Compound 169) 4- (2-benzoyl-4-chlorophenoxy) butyric acid: LC-MS analysis: m + 1 peak detected (319). Results of 1 H NMR analysis: (d6-DMSO): δ 11.9, bs, 1H (COOH); δ 7.64, d, 2H, (phenyl H, o to CO); δ 7.59, t, 1H (phenyl H, p to CO); δ 7.51, dd, 1H (aryl H, p to CO); δ 7.45, t, 2H (phenyl H, m to CO); δ 7.36, d, 1H (aryl H, o to CO); δ 7.14, d, 1H (aryl H, m to CO); δ 3.87, t, 2H, (CH ₂ α to O); δ 1.84, t, 2H (CH ₂ α to COOH); 51.53, p, 2H (median CH ₂ in the chain). ¹³ C NMR (d6-DMSO): 194.37, 173.82, 154.74, 136.96, 133.42, 131.56, 130.05, 128.97, 128.62, 128.29, 124.48, 114.61, 67.38, 29.37, 23.79.

[338] (Compound 170) 4- (2-benzoyl-4-bromophenoxy) butyric acid: LC-MS analysis: m + 1 peak detected (363). Results of 1 H NMR analysis: (d6-DMSO): δ 11.9, bs, 1H (COOH); δ 7.6, m, 3H, (aryl H); δ 7.60, t, 1H (phenyl H, p to CO); δ 7.49, dd, 1H (aryl H, o to CO); δ 7.46, t, 2H (phenyl H, m to CO); δ 7.09, d, 1H (aryl H, m to CO); δ 3.89, t, 2H, (CH ₂ α to O); δ 1.82, t, 2H (CH ₂ α to COOH); δ 1.53, p, 2H (median CH ₂ in the chain). ¹³ C NMR (d6-DMSO): 194.28, 173.81, 155.19, 136.97 134.48, 133.42, 131.06, 130.48, 128.97, 128.62, 115.08, 112.02, 67.33, 29.35, 23.77.

[339] (Compound 171) 4- (2- (2-chlorobenzoyl-5-methylphenoxy) butyric acid: LC-MS analysis: m + 1 peak detected (333). 1H NMR analysis: (d6-DMSO): δ 12.0, bs, 1H (COOH); δ 7.54, d, 1H, (aryl H); δ 7.4, m, 2H (aryl H); δ 7.33, dt, 1H (aryl H); δ 7.29, d, 1H (aryl H); δ 6.86, m, 2H (aryl H, o to O); δ 3.77, t, 2H, (CH ₂ α to O); δ 2.31, s, 3H, CH ₃ ); δ 1.85, t, 2H (CH ₂ α to COOH); δ 1.35, p, 2H (median CH ₂ in 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.

[340] (Compound 172) 4- (2- (2-chlorobenzoyl-4-methylphenoxy) butyric acid: LC-MS analysis: m + 1 peak detected (333). 1H NMR analysis: (d6-DMSO): δ 11.95, bs, 1H (COOH); δ 7.43, m, 3H, (aryl H); δ 7.34, m, 3H (aryl H); δ 6.92, d, 1H (aryl H, o to O); δ 3.74 , t, 2H, (CH ₂ α to O); δ 2.25, s, 3H, CH ₃ ); δ 1.84, t, 2H (CH ₂ α to COOH); δ 1.33, p, 2H (median CH ₂ in the chain). ¹³ NMR (b6-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.

[341] (Compound 173) 4- (2-benzoyl-4-chloro-5-methylphenoxy) butyric acid: LC-MS analysis: m + 1 peak detected (333). Results of 1H NMR analysis: (d6-DMSO): δ 11.9, bs, 1H (COOH); δ 7.61, d, 2H, (phenyl H, о to СО); δ 7.57, t, 1 N (phenyl H, p to CO); δ 7.44, t, 2H (phenyl H, m to CO); δ 7.33, s, 1H (aryl H, o to CO); δ 7.14, s, 1H (aryl H, m to CO); δ 3.87, t, 2H, (CH ₂ α to O); δ 2.33, s, 3H, CH ₃ ); δ 1.81, t, 2H (CH ₂ α to COOH); δ 1.49, p, 2H (median CH ₂ in the chain). ¹³ C NMR (d6-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.

A possible method of preparing compound F

[342] Compound F can be prepared differently using the Friedel-Crafts aromatic acylation method:

[343] Phenol suitably substituted is used, mixed with the corresponding bromoester, K ₂ CO ₃ is used as the base, the product is reacted with the corresponding aromatic acid chloride in the presence of AlCl ₃ ; or take salicylic acid, suitably substituted, and mix it with the corresponding bromoester, using K ₂ CO ₃ as the base. The product is converted to acid chloride SOCl ₂ , which then reacts with benzene, suitably substituted, in the presence of AlCl ₃ .

Example 8 - Preparation of Compound 174

[344] Compound 174 was prepared in three stages:

A. O-acetyl-5-chlorosalicylic acid

[345] 10 g (57.9 mmol) of 5-chlorosalicylic acid was weighed in a 100 ml round bottom flask, then acetic anhydride (12.8 ml, 115.9 mmol) was added. Before adding concentrated sulfuric acid (2 drops), the mixture was stirred for 5 minutes. The reaction mixture was irrigated 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. Purity was checked by HPLC, which allowed the presence of impurities to be detected. In a 200 ml round bottom flask, the precipitate was stirred overnight in water (150 ml). Insoluble solids were collected by vacuum filtration. The presence of impurities was checked by HPLC, which showed that there were no impurities in the crude product. The product was dried overnight in vacuo to give 12 g of o-acetyl-5-chlorosalicylic acid (56 mmol, 97% yield).

B. O-acetyl-5-chlorosalicyloyl chloride

[346] Thionyl chloride (~ 100 ml) was placed in 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 chilled thionyl chloride. DMF (2 drops) was added to the reaction mixture to dissolve the acid. The reaction mixture was stirred overnight to obtain a homogeneous solution. Excess thionyl chloride was distilled off in vacuo. The remaining residue was dried in vacuo overnight.

C. 3- (N-2-hydroxy-5-chlorobenzoyl) aminopropionic acid

[347] (3-Alanine (2.5 g, 28.0 mmol) was weighed in a 250 ml round bottom flask. Methylene chloride (100 ml) was added to the flask and the mixture was stirred for 5 minutes. Then, chlorotrimethylsilane was added dropwise to the flask ( 6.06 g, 55.7 mmol.) During irrigation, the reaction mixture was heated for 15 minutes. 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 into a chilled flask O-Acetyl-5-chlorosalicyloyl chloride (7.6 g, 27.9 mmol) was dissolved in methylene chloride (30 ml) and added to the reaction mixture over e 0.5 hours. The reaction mixture was stirred overnight and allowed to warm to room temperature. The reaction was confirmed by HPLC. The solvent was evaporated in vacuo. The remaining residue was stirred in 2 N NaOH (~ 100 ml) for 2 hours and slowly heated to 60 ° C. Then the solution was cooled to room temperature and filtered by a pressureless filtration method.The filtrate was slowly acidified with concentrated HCl until a precipitate formed. The crude product was collected at a pH of 6. The product was recrystallized using a MeOH-H ₂ O mixture. Purity was checked by HPLC, which showed the presence of impurities. The product was subjected to purification and recrystallization several times until a pure compound was obtained. The final product was stirred overnight in methylene chloride, collected by filtration, and dried under vacuum overnight to obtain a pinkish powder (3.98 g, 16.3 mmol, 58.5% yield); melting point 181-183 ° C; 1 H-NMR (DMSO-d6) δ 2.47-2.58 (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). The value of KF = 1.615%. The calculation results of C ₁₀ H ₁₀ NO ₄ Cl * 0.2220H2O: C, 48.50, H, 4.25, N, 5.66. Found: C, 48.20; H, 4.03; N, 5.43.

Example 9 - Preparation of Compound 175-178

Compound 175 -4- (2-benzyloxy-phenoxy) -butyric acid:

[346] In a 250-ml flask equipped with a reflux condenser and a magnetic stirrer, 2-benzyloxyphenol (8.0 g, 40 mmol), ethyl 4-bromobutanoic acid (5.7 ml, 40 mmol), and potassium carbonate were added in an inert atmosphere. (7.2 g, 52 mmol), and ethanol (100 ml). Under irrigation, the reaction mixture was heated with stirring for 8 hours. Then the reaction mixture was cooled to room temperature and the insoluble by-product was removed by vacuum filtration. 2N was added to the filtrate. aqueous sodium hydroxide solution (30 ml). This solution was heated to 50 ° C for 2 hours. Then, the solution was cooled to room temperature, ethanol was removed under reduced pressure, and the pH of the resulting solution was adjusted to 9. The aqueous solution was washed with ethyl acetate (2 × 30 ml), and the remaining ethyl acetate was removed under reduced pressure. The solution was cooled to 0 ° C. using an external ice bath, and then acidified to pH = 2–6 N. aqueous hydrochloric acid. The precipitated product was collected by vacuum filtration and dried under vacuum. The product (7.2 g, 63%) was isolated as a white powder. 1 H NMR (400 MHz, DMSO-d6): δ 12.0, s, 1H (COOH); δ 7.4, multiplet, 5H (benzyl aryl H); δ 7.0, multiplet, 2H (aryl H); δ 6.9, multiplet, 2H (aryl H); δ 5.0, s, 2H (benzyl CH ₃ ); δ 4.0, t, 2H (CH ₂ α to phenoxy); δ 2.4, t, 2H (CH ₂ α to COOH); δ 1.9, multiplet, 2H (remaining CH ₂ group).

[347] Compound 176 - (4-Benzyloxy-phenoxy) -acetic acid was purchased from Lancaster.

[348] Compound 177 - 11- (2-Benzyloxyphenoxy) undecanoic acid:

[349] Freshly ground potassium hydroxide (4.2 g, 74.91 mmol) and 100 ml of dimethyl sulfoxide were placed in a 250 ml Erlenmeyer flask. 2-Benzyloxyphenol (5 g, 24.97 mmol) and 11-bromundecanoic acid methyl ester (7 g, 25.07 mmol) were added and the mixture was stirred at room temperature overnight. Then water (75 ml) was added and the solution was heated to 85 ° C with stirring for three hours. The reaction mixture was cooled to room temperature and acidified with concentrated hydrochloric acid to pH = 2. The acidified solution was cooled to 4 ° C for 2 hours, and then the precipitate was collected by vacuum filtration. The product was recrystallized from ethanol / water. The product (8.88 g, 93%) was isolated as a light brown solid with a melting point of 62-63 ° C. Organic elemental analysis: Found: C 74.71; H 8.08%; C ₂₄ H ₃₂ O ₄ requires C 74.97; H 8.39%;

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

[351] In a 500-ml three-neck flask equipped with a reflux condenser, 4-benzyloxyphenol (30.64 g, 150 mmol), ethyl 5-bromopentanoic acid (31.99 g, 150 mmol), potassium carbonate were placed in an inert atmosphere (22.80 g, 165 mmol), and 270 ml of 2-butanone. Under irrigation, the reaction mixture was heated for 23 hours, cooled and then diluted with ethyl acetate (150 ml) and extracted with water (500 ml). The organic layer was washed with water (1 × 250 ml) and brine (1 × 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. White crystals were dissolved in ethyl acetate (100 ml), washed with aqueous 1 N. sodium hydroxide solution (3 × 50 ml), and the solvent was removed under reduced pressure. The resulting oil was dried in vacuo overnight to give white crystals. The product was recrystallized from 1: 1 ethanol / water, collected by vacuum filtration and dried under vacuum. This product was used without further purification.

[352] 5- (4-Benzyloxy-phenoxy) pentanoic acid ethyl ester (15.13 g, 46 mmol) and 2 N were placed in a 1 L round bottom flask equipped with a reflux condenser. aqueous sodium hydroxide solution (47 ml). The mixture was stirred for 30 minutes. Water (200 ml) was added. The mixture was stirred for 20 minutes, then heated under irrigation for 2 hours, forming a brown solution. By adding ice, the solution was rapidly cooled to room temperature. The cooled solution was acidified with a 2N aqueous hydrochloric acid solution (50 ml) and the white precipitate formed was collected by vacuum filtration, washed with water (2 × 100 ml), hexane (2 × 100 ml), and dried over vacuum overnight. The powder was finely ground and washed with hexane (1 × 250 ml) and diethyl ether (1 × 250 ml) to give a white powder. In a 1-liter beaker, the resulting powder was dissolved in a mixture of ethyl acetate (300 ml) and diethyl ether (200 ml). The solution was heated for 10 minutes, methanol (5 ml) was added, heated for another 10 minutes, and then filtered through a celite layer to obtain a clear yellow solution. The product crystallized by slowly adding hexane. The first batch of crystals was collected by the filtration method and hexane (200 ml) was added to the mother liquor. Then the solution was concentrated under reduced pressure to a volume of 400 ml and left for some time. A second portion of the crystals was collected by filtration and combined with the first. The product (8.92 g, 65%) was isolated as a white crystalline substance with a melting point of 127-128 ° C. Organic elemental analysis: Found: C, 71.01; H, 6.98%; C ₁₈ H ₂₀ O ₄ requires C 71.98; H 6.71%;

Compound HPLC Retention Time KF value Melting point, range CHNC C CHNC H CHNF C CHNF H 175 5.93 min 176 5.54 0 69.76 5.46 176 5.54 0 69.76 5.46 177 9.19 0 62-63 74.97 8.39 74.71 8.08 178 6.12 0 127-128 71.98 6.71 71.01 6.98

Example 10 - Oral delivery of PYY [3-361 in solid form to rats

[353] A PYY stock solution [3-36] (80 mg / ml) prepared using deionized water (PYY is available from Bachem California Inc., Torrance, California) was used.

[354] Approximately 0.08 mg / tablet (approximately 0.3 mg / kg) PYY (approximately 1 μl) was added and mixed with approximately 13.5 or 27 mg / tablet (50 or 100 mg / kg) free acid or sodium salts of the carrier compound as follows. Carver 4350 Upper Punch, Bottom Punch, and Hand Press Mold for Carver Tablets with Capsule Standard Sample by Natoli Engineering Company, Inc. (St. Charles, Missouri) was treated with magnesium stearate (0.1%). About 13.58 mg or about 27.08 mg of the mixed powder was loaded into the mold and a pellet mini-tablet was produced at a pressure of about 6894.74 kPa. The resulting solid dosage form has approximately the size of a standard capsule 9 (diameter of approximately 2.65 mm and a length of approximately 8.40 mm) for a tablet of 27.08 mg, and for a tablet of 13.58 mg, the diameter is approximately 2.65 mm and a length of approximately 4 , 20 mm.

[355] Male Sprague Dawley rats (approximately 260 to 280 g) were not fed overnight and were then anesthetized using a standard CO ₂ -alignment method for approximately 10-30 seconds, resulting in an anesthetic condition of less than approximately 1 a minute, preferably from about 10 to 30 seconds.

[356] A dosing tube for oral administration was used. A dosing tube was inserted into the rats mouth and carefully penetrated into the rats throat and esophagus by about 8-15 cm, depending on the weight of the rat (generally about 11 cm). A solid dosage form was injected into the distal esophagus and / or stomach by pressing on the plunger of the dosing tube for oral administration.

[357] Blood samples were taken retro-orbitally, usually after 0, 15, 30, 60 and 90 minutes. Serum DYY concentrations were quantified by radioimmunoassay for PYY content [3-36] (Catalog # RK-059-02 from Phoenix Pharmaceuticals, Inc., Belmont, CA). The results obtained in animals in each group were averaged for each point in time. The maximum value of these average values (ie, the average maximum concentration of PYY [3-36] in serum ± standard deviation (SD)) is given below.

Table 1 Oral administration of PYY [3-361 in rats Carrier compound Route of administration The dose of the compound (mg / kg) Dose PYY [3-36] (mg / kg) Mean maximum serum PyY content (pg / ml) ± CO 23 - sodium salt Oral, solid dose, 1 tablet per animal one hundred 0.3 427.4 ± 258.7 121 - sodium salt Oral, solid dose, 1 tablet per animal one hundred 0.3 897.1 ± 257.3 121 free acid Oral, solid dose, 1 tablet per animal fifty 0.3 161.7 ± 148.5 174 - sodium salt Oral, solid dose, 1 tablet per animal -one hundred 0.3 675 ± 427.1 174 free acid Oral, solid dose, 1 tablet per animal one hundred 0.3 0

Example 11 - Oral delivery of PUG3-361 in liquid form to rats

[358] Dosage solutions for oral administration of a carrier compound and peptide YY residues 3-36 (PYY [3-36]) (available from Bachem California Inc. of Torrance, California) in deionized water were prepared as follows.

[359] The stock solution of PYY [3-36] (80 mg / ml) was prepared using deionized water. Compositions for oral administration containing 200 mg / kg of the carrier compound and 0.3 mg / kg of PYY in aqueous solution were prepared. A solution of compound 23 was prepared by adding one equivalent of sodium hydroxide to convert the carrier agent as a free acid into its sodium salt.

[360] Male Sprague-Dawley rats weighing between 240-320 g were not fed a maximum of 24 hours before the experiments and ketamine (44 mg / kg) and torazine (1.5 mg / kg) were injected intramuscularly before the test product was administered. Then, the test composition was orally administered to the anesthetized animals. A group of five animals was administered one of the dosing solutions. For oral administration, an 11 cm French Rusch 8 catheter was attached to a 1 ml syringe with a pipette tip. The syringe was filled with the dosed solution by drawing the solution through a catheter, which was then wiped dry. The catheter was placed in the esophagus, leaving 1 cm of the tube behind the incisors. The dosing solution was injected by pressing the plunger of the syringe.

[361] Blood samples were taken serially from the caudal artery or by puncture of the heart, usually after 0, 15, 30, 45, 60, and 90 minutes. Serum PYY concentration was quantified by radioimmunoassay for PYY content [3-36] (Catalog # RK-059-02 from Phoenix Pharmaceuticals, Inc., Belmont, CA). The results obtained in animals in each group were averaged for each time point.

The maximum value of these average values (ie, the average maximum concentration of PyY [3-36] in serum ± standard deviation (SD)) is shown below in Table 2. When only Pyy [3-36] was administered orally to the animals, the significant PyY content [ 3-36] were not found in the blood.

table 2 Oral administration of PUG3-361 (liquid) in rats Compound Route of administration The dose of the compound (mg / kg) Dose PYY [3-36] (mg / kg) The average maximum concentration of PYY in serum (pg / ml) ± CO 23 Oral administration (liquid dose) 200 0.3 788.198 ± 50.59 151 Oral administration (liquid dose) 200 0.3 801.96 ± 290.61 158 Oral administration (liquid dose) 200- 0.3 1065 ± 75.352 160 Oral administration (liquid dose) 200 0.3 370.39 ± 306.29 160 Oral administration (liquid dose) 200 0.3 631.96 ± 316.16 160 Oral administration (liquid dose) 200 0.3 705.106 ± 75.906 161 Oral administration (liquid dose) 200 0.3 340.95 ± 228.946 174
Oral administration (liquid
dose) 200 0.3 1262.26 ± 313.58

Example 12 Oral Delivery of Human Recombinant Insulin to Rats

[362] Insulin (human recombinant) was purchased from ICN Biomedicals (Aurora, Ohio) as bulk powder. To prepare the initial solutions, insulin was dissolved in deionized water (pH ~ 6.5), obtaining a concentration of 15 mg / ml. Prior to use, stock solutions were stored frozen at -20 ° C in 1.0 ml aliquots. In order to obtain a final concentration of the carrier compound in dosed solutions of 200 mg / ml, the carrier compound was dissolved in deionized water. The free acid carrier reagent was converted to the sodium salt by adding one equivalent of sodium hydroxide. The solutions were shaken, sonicated and heated and, if necessary, sodium hydroxide was added again in μl amounts to achieve uniform solubility. The pH of the solutions was adjusted between 3.5-8.5 by the addition of either perchloric acid or sodium hydroxide. Then, to obtain a final concentration of 0.5 mg / ml, an initial insulin solution (usually 66.7 μl) was added to the carrier reagent solution. After dissolution and addition of the drug, the solutions were brought to the final volume by adding deionized water.

[363] Orally, either male insulin or insulin was administered to male rats in combination with the Emisphere carrier reagent. Usually, rats were not fed for 18-24 hours before administration. For insertion, the French Rusch 8 catheter was shortened to 11 cm in length and adapted for use with a 1 ml syringe. The syringe was filled with the dosed solution, and the catheter was wiped dry to remove excess solution. A catheter was inserted into the rat's mouth and guided into the esophagus (10.0 cm). The dosed solution was injected by pressing on the plunger of the syringe while holding the rat in a standing position.

Sampling and processing: insulin

[364] In blood sampling immediately prior to each sampling, rats were rapidly (~ 10 sec) exposed to carbon dioxide until they fell into a state of prostration. For blood sampling, a 77 mm capillary tube was inserted into the retro-orbital cavity. Typically, blood samples were taken prior to administration (time 0) and 15, 30, 45, and 60 minutes after administration. Samples were collected in CAPIJECT® tubes (Terumo Corporation, Tokyo, Japan) containing a blood coagulation activator (red cap, serum separation tubes). Samples were allowed to coagulate for ~ 20 minutes at 4 ° C. After coagulation, in order to separate the serum, the samples were centrifuged at 10,000 rpm for 4 minutes at 6 ° C. Serum was collected in Eppendorf tubes and cooled at −20 ° C. until analysis began.

Sampling and processing: glucose in whole blood

[365] In order to determine the pharmacodynamic response, a blood glucose meter (OneTouch Ultra, LifeScan® (Johnson & Johnson, NJ) was used to measure glucose in whole blood after administration of insulin or insulin with a carrier reagent. Samples were taken either from the retro-orbital cavity (see Sampling and processing of samples: insulin) or from the caudal artery (i.e. by cutting the tail). When cutting the tail, the tip of the tail was cut off with a scalpel by approximately 5 mm. After dropping the first drops of blood, a small sample (~ 5-10 μl) was touched on the test strip of the glucometer (OneTouch Ultra, LifeScan) and the blood glucose reading appeared on the device. At each subsequent sampling, the blood clot formed at the tip of the tail was destroyed and a fresh sample was taken. In general, samples were taken prior to administration (time 0) and 15, 30, 45, and 60 minutes after administration.

Table 3 Oral administration of insulin (liquid dose) in rats. Compound Route of administration The dose of the connection carrier (mg / kg) Insulin dose (mg / kg) Maximum% drop in glucose ± CO (control) 24 Oral administration (liquid dose) 200 0.5 -14.73 ± 17.64 25 Oral administration (liquid dose) 200 0.5 -14.81 ± 12.99 26 Oral administration (liquid dose) 200 0.5 -25.93 ± 14.86 27 Oral administration (liquid dose) 200 0.5 -25.40 ± 30.61 28 Oral administration (liquid dose) 200 0.5 -11.41 ± 18.92 29th Oral administration (liquid dose) 200- 0.5 -29.25 ± 6.97 140 Oral administration (liquid dose) one hundred one Smah = 81.16 ± 114.98 μIU / ml 141 Oral administration (liquid dose) one hundred one Smah = 204.05 ± 60.88 μIU / ml 142 Oral administration (liquid dose) one hundred one Smah = 118.16 ± 72.75 μIU / ml 145 Introduction through the colon fifty 0.1 Smah = 15.03 ± 7.80 μIU / ml 145 Oral administration (liquid dose) one hundred one Amp = 3.92 ± 5.62 μIU / ml

160 Oral administration (liquid dose) 200 0.5 Amp = 74 ± 7.5 μIU / ml 165 Oral administration (liquid dose) 200 0.5 -33.0 166 Oral administration (liquid dose) 200 0.5 -5.7 167 Oral administration (liquid dose) 200 0.5 -21.2 167 Oral administration (liquid dose) 200 0.5 -17.7 167 Oral administration (liquid dose) 200 0.5 -26.2 167 Oral administration (liquid dose) 200 0.5 -17.8 167 Oral administration (liquid dose) 200 0.5 -22.7

Example 13 - Human Zinc Insulin - Oral Delivery

[366] The compositions of the carrier compound and human zinc insulin for oral administration (minimum 26 IU / mg, available from Calbiochem - Novabiochem Corp, La Jolla, California) were prepared in deionized water. 500 mg of the carrier compound was added to 1.5 ml of water. The free acid included in the carrier compound was converted into the sodium salt by stirring the resulting solution and adding one equivalent of sodium hydroxide. The solution was shaken, then heated (approximately 37 ° C.) and sonicated. The pH was adjusted in the range of about 7-8.5 with NaOH or HCl. To achieve uniform solubility, additional amounts of NaOH were added, if necessary, and the pH was again adjusted to between about 7 and 8.5. Then, the total volume was adjusted to about 2.4 ml with water and shaken. Approximately 1.25 mg of insulin from the initial insulin solution (15 mg / ml, prepared by mixing 0.5409 g of insulin and 18 ml of deionized water, adjust pH = 8.15 with HCl and NaOH and, to obtain a clear solution, use 40 ml of concentrated HCl, 25 ml of 10 N NaOH and 50 ml of 1 N NaOH) were added to the solution and stirred by inverting. The final doses of the carrier compound, insulin and dose volumes are given.

[367] Male Sprague-Dawley rats weighing approximately 200-250 g were not fed for 24 hours and ketamine (44 mg / kg) and chlorpromazine (1.5 mg / kg) were administered 15 minutes before administration and again, preserve anesthesia if necessary. A group of five animals was administered one of the dosing solutions. For oral administration, an 11 cm French Rusch 8 catheter was attached to a 1 ml syringe with a pipette tip. The syringe was filled with the dosed solution by drawing the solution through a catheter, which was then wiped dry. The catheter was placed in the esophagus, leaving 1 cm of the tube behind the incisors. The dosing solution was injected by pressing the plunger of the syringe.

[368] Blood samples were taken serially from the caudal artery, generally after 15, 30, 60, 120 and 180 minutes. Serum insulin levels were determined using the Insulin ELISA test kit (kit # DSL-10-1600 from Diagnostic Systems Laboratories, Inc., Webster, Texas), adjusting the standard protocol to optimize the sensitivity and linear region of the calibration curve for sample volumes and concentrations used in this protocol. Serum human insulin concentrations (μU / ml) were measured for each point in time for each of the five animals in each group. Five values for each point in time were averaged and the results were plotted: insulin concentration - time.

(Previous experiments did not reveal measurable levels of human insulin after administration without a carrier reagent). The maximum (peak) and area under the curve (PPC) are shown in Table 4 below. For% change from baseline for Blood Glucose the ONE TOUCH® (Life Scan, Johnson & Johnson, New Brunswick, NJ).

Table 4
Insulin - Oral Delivery Carrier compound The dose of the connection carrier (mg / kg) Insulin dose (mg / kg) Volumetric dose (ml / kg) Amp (glucose) (µU / ml) PPK 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 - Delivery Through the Lungs

[368] Dosage compositions of a carrier compound and human insulin in water were prepared. In general, deionized water was added to 1.5 mg of the carrier compound, bringing the volume to 1.0 ml, and the solution was shaken. Either the sodium salt of the carrier compound was used, or the free acid was converted into the sodium salt by stirring the resulting solution, adding one equivalent of sodium hydroxide (10 N) and diluting with water. The solution was shaken, then heated (approximately 37 ° C.) and sonicated. The pH was adjusted from about 7.0 to 8.5 with NaOH or HCl. 75 μl of human insulin stock solution (2 mg / ml) was added to the solution. (A stock solution was prepared as follows. To 0.02 g of insulin was added 3 ml of a solution of HCl in deionized water with a pH of 3.0. The pH of the resulting solution was adjusted with HCl and NaOH to below 3.0 (approximately 2.6) to until the solution became clear. Then the pH was adjusted to 7.6 using NaOH and HCl. The final volume was adjusted to 10 ml with deionized water with a pH of 7.5. Final pH = 7.59). Then water was added, bringing the total volume to 2.0 ml, and the solution was carefully inverted several times. According to the administration protocol, the solution can be used immediately after preparation, or in another example implementation, before administration it can be placed in a water bath with a temperature of 37 ° C for one hour. The final dose of the carrier compound, the dose of insulin and the dose volumes are shown below in table 4.

[369] Typical injection and sampling protocols are as follows. Male Sprague-Dawley rats weighing between 200-250 g were not fed for 24 hours, and ketamine (44 mg / kg) and chlorpromazine (3.0 mg / kg) were administered 15 minutes before administration and, if necessary, maintain anesthesia, they were administered again (using the same amounts of ketamine and 1.5 mg / kg of chlorpromazine). Typically, a group of five animals was administered one of the dosing solutions. A control group of five animals was injected with insulin separately. A tracheal dropper for rodents equipped with a light source (available from Perm Century, Inc., Pittsburgh, PA) was filled with a dosing solution and inserted into the pharynx so that the needle entered the trachea (visually confirmed). The dosed solution was introduced by pressing the piston.

[370] Blood samples from each animal were taken serially from the caudal artery, generally 5, 15, 30, 60, and 120 minutes after administration. Serum insulin levels were determined using the Insulin ELISA test kit (kit # DSL-10-1600 from Diagnostic Systems Laboratories, Inc., Webster, Texas), adjusting the standard protocol to optimize the sensitivity and linear region of the calibration curve for sample volumes and concentrations used in this protocol. Serum insulin concentrations (μU / ml) were measured for each point in time for each of the five animals in each group. Five values for each point in time were averaged and the results were plotted: insulin concentration - time. The ratio of the area under the curve (PPC) for the test group relative to the area of the control group is given below. The ratio of the maximum serum insulin concentration (Cmax) in the test group relative to the maximum concentration in the control group is also given below.

Table 5 Delivery of insulin through the lungs Carrier compound Volumetric dose (ml / kg) The dose of the connection carrier (mg / kg) Insulin dose (mg / kg) Smah 174 0.4 16 0.03 18.36 ± 19.18

Example 15 - Delivery of heparin orally and through the colon

[371] Dosage solutions for oral administration and administration through the colon (BTK) containing a carrier compound and sodium heparin USP in a 25% aqueous propylene glycol solution were prepared. The sodium salt of this compound was used. In general, the carrier compound and heparin in the form of dry powders (about 166-182 IU / mg) were mixed with shaking. This dry mixture was dissolved in a 25% (by volume) aqueous propylene glycol solution, shaken, and placed in an ultrasonic treatment device (approximately 37 ° C). The pH was adjusted to about 7 (6.5 to 8.5) using an aqueous solution of NaOH (2 N). The dosing solution was sonicated to obtain a clear solution. The final volume was adjusted to 3.0 ml. The final dose of the carrier compound, the dose of heparin and dose volumes are shown in table 6.

[372] The following typical protocols for administration and sampling were used. Male Sprague-Dawley rats weighing 275-350 g were not fed for 24 hours and were anesthetized with ketamine hydrochloride (88 mg / kg) by intramuscular injection immediately prior to administration. A group of five animals was administered one of their dosing solutions. For oral administration of 11 cm, the Rusch 8 French catheter was attached to a 1 ml syringe with a pipette tip. The syringe was filled with the dosed solution by drawing the solution through a catheter, which was then wiped dry. The catheter was placed in the esophagus, leaving 1 cm of the tube behind the incisors. The solution was injected by pressing the syringe plunger. When introduced through the colon (VTK) 7.5 cm, the French Rusch 8 catheter was attached to a 1 ml syringe with the tip of a pipette. A dosing catheter was inserted into the colon through the anus until the tube was no longer visible. The dosed solution was slowly injected into the colon.

[373] Citrate blood samples were taken by heart puncture, previously injected with ketamine (88 mg / kg), usually after 0.25, 0.5, 1.0 and 1.5 hours. Heparin activity was determined using partial thromboplastin activation time (HAT) according to the method of Henry, J.B., Clinical Diagnosis and Management by Laboratory Methods, Philadelphia, PA, W.B. Saunders (1979). According to preliminary studies, the baseline values are approximately 20 seconds. The results obtained in the study of five rats in each group were averaged for each point in time. The resulting maximum is shown below in table 6.

Table 6 Oral / Intestinal Delivery of Heparin Carrier compound Method of administration Volumetric dose (ml / kg) The dose of the compound (mg / kg) Dose of heparin (mg / kg) Average maximum RFID (sec) ± SD) pH 114 VTK one fifty 25 42.90 ± 8.70 7.61 140 VTK one fifty 25 23.49 ± 6.12 7.67 141 VTK one fifty 25 52.40 ± 21.54 7.62 143 VTK one fifty 25 114.69 ± 121.62 7.18 145 VTK one fifty 25 134.42 ± 99.03 6.93 151 Oral 3 300 one hundred 252.09 ± 107.13 151
VTK one fifty 25 2.36 ± 1.27 (antifactor
Ha) 7.06 151 VTK one 25 25 3.39 ± 3.07 (antifactor Xa) 7.23 160 VTK one fifty 25 131 + 154 (Tmax = 90 min)

Example 16 - Delivery of parathyroid hormone (PTH 1-34) - Oral / intracolic delivery

[374] Solutions of a carrier compound and residues of human parathyroid hormone 1-34 (PTH) in water were prepared for oral and / or colon administration. The sodium salt of the carrier compound was used. Typically, a solution of this compound was prepared in water and mixed by adding one equivalent of sodium hydroxide (1.0 N) to obtain the sodium salt. Final dosing solutions were prepared by mixing the compound with a PTH stock solution (PTH was obtained from Eli Lilly and Co., Indianapolis, Indiana) (usually having a concentration of 5 mg PTH / ml) and diluting to the required volume (usually 3.0 ml). The final compound, PTH and volumetric amounts of doses are shown below in table 7.

[375] The following typical injection and sampling protocols were used. Male Sprague-Dawley rats weighing 200-250 g were not fed for 24 hours and ketamine (44 mg / kg) and chlorpromazine (1.5 mg / kg) were administered 15 minutes before administration. A group of five rats was administered one of the dosing solutions. For oral administration of 11 cm, the Rusch 8 French catheter was attached to a 1 ml syringe with a pipette tip. The syringe was filled with the dosed solution by drawing the solution through a catheter, which was then wiped dry. The catheter was placed in the esophagus, leaving 1 cm of the tube behind the rat incisors. The solution was injected by pressing the syringe plunger. When introducing through the colon (VTK) 7.5 cm, the French Rusch 8 catheter was attached to a 1 ml syringe with the tip of an Eppendorf pipette. The syringe was filled with the dosed solution by pulling the solution through a catheter tube. The catheter tube was wiped dry. The pipette tip was smeared with K-Y jelly gel, avoiding contact with the ear of the tube, and the tube was inserted into the colon until it was visible. The solution was injected by pressing the syringe plunger and the tube was removed.

[376] Blood samples were taken serially from the caudal artery, generally after 0, 15, 30, 45, 60, and 90 minutes when administered orally and 0, 10, 20, 30, 60, and 90 minutes when introduced through the colon. Serum PTH concentration was quantified using a PTH radioimmunological assay kit (Kit # RIK 6101 from Peninsula Laboratories, Inc. San Carlos, CA). According to preliminary studies, the baseline values are approximately zero. The blood test results of five rats in each group were averaged for each point in time. The maximum value is shown below in table 7.

Table 7 PTH delivery to rats via the oral route or through the colon Carrier compound Route of administration Volumetric dose (ml / kg) The dose of the compound (mg / kg) PTH dose (m kg / kg) The average maximum concentration [PTH] in serum (PG / ml) ± CO pH 113 Oral one one hundred 200 780.77 ± 439.92 8.18 113 Oral one one hundred 200 53.51 ± 39.55 8.09 114 Oral one one hundred 200 135.78 ± 136.97 8.41

Example 17 - Interferon - Oral Delivery

[377] In deionized water, metered solutions of the carrier compound and interferon alfacon-1 (IFN) were prepared (available as Infergen® from InterMune, Inc. Brisbane, California). The free acid included in the carrier compound was converted to the sodium salt by adding one equivalent of sodium hydroxide. Typically, a solution of the carrier compound in water was prepared and mixed by adding one equivalent of sodium hydroxide (1.0 N) to obtain the sodium salt. This mixture was shaken and placed in an ultrasonic treatment device (approximately 37 ° C). The pH was adjusted from about 7.0 to 8.5 by adding an aqueous solution of NaOH. To obtain a homogeneous suspension or solution, the mixture was shaken, and, if necessary, an ultrasonic treatment device and heating were used. To achieve uniform solubility, an additional amount of NaOH was added, if necessary, and the pH was again adjusted in the range of 7.0 to 8.5. A solution of the carrier compound was mixed with the stock INF solution (approximately 22.0-27.5 mg / ml in phosphate buffered saline) and diluted to the desired volume (usually 3.0 ml). The final dose of the carrier compound and INF and dosage volumes are shown in Table 8.

[378] The following protocols have been commonly used for injection and sampling. Male Sprague-Dawley rats weighing approximately 200-250 g were not fed for 24 hours and ketamine (44 mg / kg) and chlorpromazine (1.5 mg / kg) were administered 15 minutes before administration and, if necessary, maintain anesthesia, introduced them again. A group of five animals was administered one of the dosing solutions. An 11 cm French Rusch 8 catheter was attached to a 1 ml syringe with the tip of a pipette. The syringe was filled with the dosed solution by drawing the solution through a catheter, which was then wiped dry. The catheter was placed in the esophagus, leaving 1 cm of the tube behind the incisors. The dosing solution was injected by pressing the plunger of the syringe.

[379] Blood samples were taken serially from the caudal artery, usually after 0, 15, 30, 45, 60, and 90 minutes. The concentration of INF was quantified using an immunological kit with a cytoscreen for the determination of human alpha-INF (catalog # KHC4012 from Biosource International, Camarillo, California). According to preliminary studies, the baseline values were approximately zero. The results obtained by analyzing the blood of animals from each group were averaged for each point in time. The maximum of these values (for example, the average maximum concentration of INF in serum) is shown below in table 8.

Table 8 Interferon - Oral Delivery Carrier compound The dose of the connection carrier (mg / kg) INF dose (mg / kg) Volumetric dose (ml / kg) The average maximum concentration of INF in serum (ng / ml) ± CO pH 141 200 one one 0.73 ± 0.44 8.29 147 200 one one 1.27 ± 0.60 8.45 174 200 one one 0.5 ± 0.57 174 200 one one 0.18 ± 0.17 174 200 one one 3.96 ± 2.72 174 200 one one 17.4 ± 9.12

Example 18 - Oral delivery of salmon calcitonin (LKT)

[380] Compositions were prepared for oral administration of a carrier compound and salmon calcitonin (LKT) in water. Typically, 450 mg of the carrier compound was added to 2.0 ml of water. Either the sodium salt of the compound was used, or the free acid was converted to the sodium salt by stirring the resulting solution, adding one equivalent of sodium hydroxide (1.0 N) and diluting with water. The solution was shaken, then heated (approximately 37 ° C.) and sonicated. The pH was adjusted to about 7 (6.5 to 8.5) by adding NaOH or HCl. 90 g of LKT from the stock solution was added to the solution. Then, the total volume was adjusted to about 3.0 ml with water (varies depending on the solubility of the carrier compound). The final dose of the carrier compound, the dose of LKT and the dose volumes are shown below in table 9.

[381] Male Sprague-Dawley rats weighing approximately 200-250 g were not fed for 24 hours and ketamine (44 mg / kg) and chlorpromazine (1.5 mg / kg) were administered 15 minutes prior to administration. A group of five animals was administered one of the dosing solutions. For oral administration of 11 cm, the Rusch 8 French catheter was attached to a 1 ml syringe with a pipette tip. The syringe was filled with the dosed solution by drawing the solution through a catheter, which was then wiped dry. The catheter was placed in the esophagus, leaving 1 cm of the tube behind the rat incisors. The dosing solution was injected by pressing the plunger of the syringe.

[382] Blood samples were taken serially from the caudal artery, usually after 0, 10, 20, 30, 60, and 90 minutes. Serum LKT concentrations were determined by testing using the EIA kit (Kit # EIAS-6003 from Peninsula Laboratories, Inc., San Carlos, California), adjusting the standard protocol from the kit as follows: kept with a mixture of 50: 1 peptide: antibody for 2 hours with shaking in the dark, rinse the cup, add serum and biotinylated peptide and dilute 4 ml of buffer, and shake in the dark overnight. The indicators were set in accordance with the values of the baseline obtained at time = 0. The blood test results of five rats in each group were averaged for each point in time. The maximum is shown in table 9 below.

Table 9 Oral delivery of salmon calcitonin (LKT) Carrier compound Volumetric dose (ml / kg) The dose of the compound (mg / kg) LHC dose (mg / kg) The average maximum concentration of LKT in serum (PG / ml ± CO) (SE) 174 150 thirty one 182.83 ± 184.82 174 150 thirty one 198.21 ± 205.15 174 150 thirty one 70.81 ± 118.47

Example 19 Delivery of Recombinant Human Growth Hormone (rhGH) Orally or Through the Colon

[383] Solutions of the carrier compound and rhGH were prepared in phosphate buffer for oral and / or colon entry (rhGH can be purchased from Novartis, Basel, Switzerland). The sodium salt of the carrier compound was obtained by reaction of the free acid with one equivalent of sodium hydroxide. Final dosing solutions were prepared by mixing the compound with the rhGH stock solution (15 mg rhGH / ml) and diluting to the desired volume (usually 3.0 ml). Dosage amounts of compounds and rhGH are shown below in table 10.

[384] The following typical protocols for administration and sampling were used. Male Sprague-Dawley rats weighing 200-250 g were not fed for 24 hours and ketamine (44 mg / kg) and chlorpromazine (1.5 mg / kg) were administered 15 minutes before administration. A group of five rats was administered one of the dosing solutions. For oral administration of 11 cm, the Rusch 8 French catheter was attached to a 1 ml syringe with a pipette tip. The syringe was filled with the dosed solution by drawing the solution through a catheter, which was then wiped dry. The catheter was placed in the esophagus, leaving 1 cm of the tube behind the rat incisors. The solution was injected by pressing the syringe plunger. When introducing through the large intestine (BTK) 7.5 cm, Rusch 8 catheter tubes (French 8 or 6) were attached to a 1 ml syringe with the tip of an Eppendorf pipette. The syringe was filled with the dosed solution by pulling the solution through a catheter tube. The catheter tube was wiped dry. The pipette tip was smeared with K-Y jelly gel, avoiding contact with the ear of the tube, and the tube was inserted into the colon through the anus until it was invisible. The solution was injected by pressing the syringe plunger and the tube was removed.

[385] Blood samples were taken serially from the caudal artery or retro-orbital cavity, usually after 0, 15, 30, 45, 60, and 90 minutes when administered orally and 0, 10, 20, 30, 60, and 90 minutes when administered intracranially. Samples were collected in CAPIJECT® tubes (Terumo Corporation, Tokyo, Japan) containing a coagulant activator (red cap, serum separation tubes). Samples were allowed to coagulate for ~ 20 min at 4 ° C. Serum rhGH concentrations were quantified using an rhGH immunoassay assay kit (Kit # K1F4015 from Genzyme Corporation Inc., Cambridge, Mass.). Five samples for each time period were combined. According to preliminary studies, the baseline values were approximately equal to zero.

[386] The maximum concentrations for each group are shown below in table 10.

Table 10 Oral / intrathecal delivery of rhGH to rats Carrier compound Route of administration Volumetric dose (ml / kg) The dose of the compound (mg / kg) Dose of rhGH (mg / kg) The average maximum concentration of rhGH in serum (ng / ml) 160 Oral one 200 3 - 161 Oral one 200 3 1.033 (± 2.31) (Tmax = 15min) 174 Oral one 200 3 57.42

All publications, references, patents, published patent applications that are referred to herein are referenced.

Claims (17)

1. A compound selected from:

and their pharmaceutically acceptable salts. 2. Pharmaceutical composition for the delivery of biologically active substances, containing:
(A) at least one biologically active substance selected from the peptide YY [PYY], insulin and parathyroid hormone; and
(B) an effective amount of a compound selected from

and their pharmaceutically acceptable salts. 3. The pharmaceutical composition according to claim 2, characterized in that said biologically active substance is human zinc insulin. 4. The pharmaceutical composition according to claim 2, characterized in that the biologically active substance is PYY [3-36]. 5. The pharmaceutical composition according to claim 2, characterized in that the biologically active substance is a human parathyroid hormone (PTH-1-34). 6. A method of administering a biologically active substance selected from the YY [PYY] peptide, insulin and parathyroid hormone to an animal that needs said substance, comprising orally administering to the animal a pharmaceutical composition according to claim 2. 7. A compound selected from the following:

and their pharmaceutically acceptable salts. 8. A pharmaceutical composition for the delivery of biologically active substances, comprising:
(A) at least one biologically active substance selected from the peptide YY [PYY], insulin, heparin and interferon; and
(B) an effective amount of a compound according to claim 7. 9. The pharmaceutical composition according to claim 8, characterized in that said biologically active substance is interferon alfacon-1. 10. The pharmaceutical composition of claim 8, characterized in that the biologically active substance is a human recombinant insulin. 11. The pharmaceutical composition according to claim 8, characterized in that the biologically active substance is heparin. 12. The pharmaceutical composition according to claim 8, characterized in that the biologically active substance is PYY [3-36]. 13. The method of introducing a biologically active substance selected from the peptide YY [PYY], insulin, heparin and interferon, an animal that needs the specified substance, including oral administration to the specified animal pharmaceutical composition according to claim 8. 14. The compound is:

and its pharmaceutically acceptable salts. 15. A pharmaceutical composition for the delivery of biologically active substances, comprising:
(A) insulin and
(B) an effective amount of a compound according to claim 14. 16. The pharmaceutical composition according to clause 15, wherein said insulin is a human recombinant insulin. 17. A method of administering insulin to an animal that needs said insulin, comprising orally administering to the animal a pharmaceutical composition according to claim 15.