# Patent

## Title
Compounds to treat diabetes and associated conditions

## Abstract

Compounds are provided that lower blood glucose concentrations, lower serum triglyceride concentrations, lower systolic blood pressure, and increase glucose uptake by adipose tissue, but do not affect the expression of PPAR-γ by adipose tissue.

## Background
CROSS REFERENCE TO RELATED APPLICATION
This application is a divisional application based on U.S. Ser. No. 09/436,047, filed Nov. 8, 1999, now U.S. Pat. No. 6,525,093 which issued on Feb. 25, 2003, the entirety of which is incorporated herein by reference.


BACKGROUND OF THE INVENTION
The present application is directed to novel antidiabetic compounds.
The causes of Type I and Type II diabetes are still unknown, although both genetic and environmental factors seem to be involved. Type I diabetes (or insulin-dependent diabetes) is an autonomic immune disease in which the responsible autoantigen is still unknown. Patients with Type I diabetes need to take insulin intravenously to survive. Type II diabetes (formerly referred to as non-insulin dependent diabetes) is a metabolic disorder resulting from the body's inability either to make a sufficient amount of insulin or to properly use the insulin that is produced. Insulin secretion and insulin resistance are considered the major metabolic defects, but the precise genetic factors involved remain unknown.
Patients with diabetes usually have one or more of the following defects:

    
    
        Under-production of insulin by the pancreas
        Over-secretion of glucose by the liver
        Defects in glucose transporters
        Desensitization of insulin receptors
        Defects in metabolic breakdown of polysaccharides
    
    


In addition to the IV administration of insulin, currently available medications used for diabetes include 4 classes of oral hypoglycemic agents listed in the following table.














Marketed
Mechanism of



Class
Drugs
Action
Limitations







Sulfonyl-
First
Signals beta cells
Development of


ureas
generation: 2
to release more
resistance



Second
insulin
Hypoglycemia



generation: 3



Bi-
Metformin
Reduces hepatic
Adverse hepatic effects


guanides

glucose
Lactic acidosis




production
Unwanted




Improves
Unwanted




Improves
gastrointestinal effects




sensitivity to




insulin


Glu-
Acarbose
Reduces glucose
Works only after meals


cosidase

absorption from
GI side effects


inhibitors

gut


Thia-
Troglitazone
Reduce insulin
Not effective in 25% of


zoli-
Rosi-
resistance
patients


dinediones
glitazone

Require frequent liver



Pia-

function tests



glitazone

Have very long onset of





action





Cause weight gain









As is apparent from the above table, there are disadvantages to each of the currently available agents for use in the treatment of diabetes. Accordingly, there is a continuing interest in the identification and development of new agents, particularly orally administered, water-soluble agents that can be used for the treatment of diabetes.
SUMMARY OF THE INVENTION
Compounds having the general formula (I)-(III) have glucose-lowering activity. 




Stereocenters (designated by *) could be R— or S—.
Each bond represented by dotted lines could be a double or a single bond, and the geometry across the bond could be E or Z.
R and R′ are independently H or C1-C20 linear or branched alkyl or alkenyl groups that may be substituted, or functional groups like COOR3, where R3═H, a cation, C1-C20 linear or branched alkyl or C5-C10 aryl; CONR1R2, where R1 and R2 may be independently or together H, linear or branched C1-C20 alkyl or C5-C20 aryl, NH2, OH, C1-C20 linear or branched alkoxy, halo, cyano, or R+R′═O.
A, A′, A′′, and C are independently H, C1-C20 acylamino, C1-C20 acyloxy, linear or branched C1-C20 alkanoyl, C3-C20 alkoxycarbonyl, C1-C20 linear or branched alkoxy; C1-C20 linear or branched alkylamino, C1-C20 alkylcarboxylamino, C1-C20 carbalkoxy; carboxyl, cyano, halo, hydroxy; and n, m, p and v are independently integers from 0 to 3;
B, B′, and B′′ are independently H, C1-C20 acylamino, C1-C20 acyloxy; C1-C20 linear or branched alkanoyl, C1-C20 linear or branched alkenyl, C1-C20 alkoxycarbonyl, C1-C20 linear or branched alkoxy; C1-C20 linear or branched alkyl amino, C1-C20 alkyl carboxyl amino, C1-C20 carbalkoxy; aroyl, araalkanoyl, carboxyl, cyano, halo, hydroxy; and q, r and s are independently integers from 0 to 3;
with the proviso that when m, n, p , q, r, s and v are zero, any unsubstituted positions on the aromatic rings are satisfied with hydrogen;
R′′, R′′′ and R′′′′ are independently H, C1-C20 linear or branched alkyl or alkenyl groups which may contain substituents, COOH, C1-C20alkoxycarbonyl, NH2, CONH2, C1-C20 acylamino, OH, C1-C20 alkoxy, halo, or cyano.
X═NH, O, S, S═O, or SO2. 




Stereocenters (designated by *) could be R— or S—.
Each bond represented by the dotted line could be a double or a single bond, and the geometry across it may be E or Z.
A, A′, and C are independently H, C1-C20 acylamino, C1-C20 acyloxy, C1-C20 alkoxycarbonyl, C1-C20 alkoxy, C1-C20 linear or branched alkyl amino, C1-C20 alkylcarboxylamino, C1-C20 carbalkoxy; carboxyl, cyano, halo, hydroxy; and t, u, and w are independently integers from 0 to 3;
B and B′ are independently H, C1-C20 acylamino, C1-C20 acyloxy; C1-C20 alkanoyl, C1-C20 alkenoyl, C1-C20 alkenyl, C1-C20 alkoxycarbonyl, C1-C20 linear or branched alkoxy, C1-C20 linear or branched alkyl amino, C1-C20 alkylcarboxylamino, C1-C20 carbalkoxy, C6-C20 aroyl, C6-C20 araalkanoyl, carboxyl, cyan, halo, hydroxy; and x and y are independently integers from 0 to 3;
with the proviso that when t, u, w, x and y are zero, any unsubstituted positions on the aromatic rings are satisfied with hydrogen;
R′, R′′, and R′′′ are independently H or C1-C20 linear or branched alkyl or alkenyl groups which may contain substituents, COOH, C1-C20 alkoxycarbonyl, NH2, CONH2, C1-C20 acylamino, C1-C20 alkoxycarbonyl, OH, C1-C20 alkoxy, halo or cyano.
X═NH, O, S, S ═O, or SO2. 




Stereocenters (designated by *) could be R— or S—.
The bond represented by the dotted line could be a double or a single bond, and the geometry across it may be E or Z.
A and C are independently H, C1-C20 acylamino, C1-C20 acyloxy, C1-C20 linear or branched alkanoyl, C1-C20 alkoxycarbonyl, C1-C20 linear or branched alkoxy, C1-C20 linear or branched alkyl amino, C1-C20 alkylcarboxylamino, C1-C20 carbalkoxy; carboxyl, cyano, halo, hydroxy; thiol, SOR or SOR2; and f and g are independently integers from 0 to 3;
B is independently H, C1-C20 acylamino, C1-C20 acyloxy; C1-C20 linear or branched alkanoyl, C1-C20 linear or branched alkenoyl, C1-C20 linear or branched alkenyl, C1-C20 alkoxycarbonyl, C1-C20 linear or branched alkoxy, C1-C20 linear or branched alkyl amino, C1-C20 alkylcarboxylamino, C1-C20 carbalkoxy, C5-C20 aroyl, C6-C20 araalkanoyl, carboxyl, cyan, halo, hydroxy; and e is an integer from 1 to 3:
with the proviso that when f and g are zero, any unsubstituted positions on the aromatic rings are satisfied with hydrogen;
R′, R′′, and R′′′ are independently H or C1-C20 linear and branched alkyl or alkenyl groups which may contain substituents, COOH, C1-C20 alkoxycarbonyl, NH2, CONH2, C1-C20 acylamino, C1-C20 alkoxycarbonyl, OH, C1-C20 alkoxy, halo, cyano.



BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 shows the blood glucose concentrations found in ob/ob mice given the representative compound at a dose of 50 mg/kg for 7 days.
FIG. 2 shows the blood glucose concentrations found in diabetic ob/ob mice given the representative compound at doses of 0 (vehicle), 10, 25, or 50 mg/kg for 7 days (left); and those found in lean ob/ob mice given the representative compound at a dose of 50 mg/kg for the same period (right).
FIG. 3 shows the serum triglyceride concentrations and systolic blood pressure of fructose-fed, insulin-resistant rats that received the representative compound or the vehicle for 7 days.
FIG. 4 shows the glucose uptake of 3T3-L1 cells exposed to two different concentrations of the representative compound (0.1 nM and 0.1 μM).
FIG. 5 shows the levels of PPAR-γ expression found in adipose tissue of ob/ob mice treated with the vehicle or the representative compound (50 mg/kg) for 10 days.



DESCRIPTION OF THE PREFERRED EMBODIMENTS
In the compounds of the formulas I, II and III, the alkyl groups may be linear or branched including but not limited methyl, ethyl, propyl, isopropyl, sec-butyl, n-butyl, pentyl, isopentyl, and the like. Alkenyl groups of 1 to 20 carbon atoms includes but is not limited to, ethylene, propylene, butylene, isobutylene, and the like. Aryl groups include phenyl, and other multi-ring aromatic structures. Alkoxy includes methoxy, ethoxy propoxy, isopropoxy, n-butoxy, isobutoxy and the like. Halo includes bromo chloro, fluoro, iodo.
Acylamino includes the group 




wherein R could be hydrogen, alkyl or aryl.

Acyloxy includes the group 




wherein R is hydrogen, alkyl or aryl.

Alkanoyl includes the group 




wherein R can be hydrogen, alkyl or aryl.

Alkoxy carbonyl includes the group 




wherein R can be alkyl.

Alkylamino includes the group 




wherein the amino group may be mono or di-substituted with alkyl groups.

Alkylcarboxylamino includes the group 




wherein R can be an alkyl group.

Carboalkoxy includes the group 




wherein R is an alkyl group.

Aroyl includes the group 




wherein R is aryl.

Araalkanoyl includes the group 




wherein R is aryl and R1 is alkylenyl.

Preferred compounds of formula are those wherein C and A are hydrogen, and q═2 when B is methyl. Other preferred compounds are those in which A′ is hydrogen and r═O, and in which A′′ is hydrogen and s═O. Another preferred class of compounds comprises those in which R is hydrogen and R′ is —COOR3. A preferred class of substituent comprises those in which R′′′′ is hydrogen, R′′′ and R′′′′ are independently —COOR3 and X is oxygen.
The method used for the synthesis of the representative member of the Type (I) compounds is shown in Scheme I. 




A specific method for the synthesis of a representative number of compounds of the formula I is shown below in Scheme IA. Compounds of the formula II where made starting from the second step showing in Scheme I with the appropriate starting materials. The compounds of formula III may be made by utilizing the chemistry of the last step of Scheme I using the appropriate starting materials. 




The compounds of the present invention are useful in the treatment of diseases or disorders characterized by the presence of the elevated blood glucose levels, that is hyperglycemic disorders, such as diabetes melitis, including both type I and II diabetes as well as other hyperglycemic related disorders such as obesity, increased cholesterol kidney related disorders, and the like. The compounds are preferably administered at least to reduce the blood glucose level in the host suffering from the hyperglycemic disorder. The sufficient amount of the compound is administered to the subject to reduced the blood glucose level to an acceptable range which is typically about plus or minus 10%, usually plus or minus 8%, and more usually plus or minus 5% of the normal average blood glucose level for the subject. A variety of hosts may be treated with the compounds to reduce blood glucose levels, such as humans and including mammalians hosts such as livestock, valuable or rare animals and pets, such as dogs and cats. The compounds may be administered by any convenient administration technique including, but not limited to, intravenous, intradermal, intramuscular, subcutaneous, or oral. The dosage delivered to the host will necessarily depend upon the route by which the compound is administered but will generally range from about 50-500 mg/70 kg human body weight, and usually from about 100-200 mg/70 kg human body weight.
The compounds will be combined in a physiologically acceptable vehicle to produce a pharmaceutical composition. The nature of the physiologically acceptable vehicle will necessarily depend on the method for which the pharmaceutically composition is administered. Exemplary vehicles include water, that is, sterile water for injection, saline, such as phosphate buffered saline, lyophilized powder in the form of tablets or capsules where such forms may include various fillers, binders and the like. The amount of the active compound in the pharmaceutical composition will be selected in view of the method by which the pharmaceutical composition is to be administered, and may be determined empirically by those of ordinary skill in the art.
FIGS. 1 through 5 present the results of preclinical tests performed using a compound according to the present invention, 4-(1-carboxy-2-(3,5-dimethoxyphenyl)) ethylenyl-4′-(2,2-dicarbomethoxy) ethyl diphenyl ether.
When 6-week-old male ob/ob mice were given a 50 mg/kg dose of this compound or the vehicle daily for 7 days, the blood glucose concentrations of the mice given the compound were reduced 50% from those of the mice given the vehicle only, and the reductions of blood glucose concentrations were observed as early as Day 2 (see FIG. 1).
In another experiment, 6-week-old male diabetic ob/ob mice received the indicated oral doses of the test compound daily. FIG. 2 shows that the 10 mg/kg dose of the compound lowered blood glucose concentrations as effectively as the 50 mg/kg dose. The blood glucose concentrations in lean control animals given the highest dose of the test compound (50 mg/kg) did not differ from those in animals given vehicle only.
The ability of this test compound to lower serum triglyceride concentrations and blood pressure was studied in fructose-fed, insulin-resistant rats. For this experiment, male Sprague-Dawley rats initially weighing 150-175 g were placed on a 60% fructose-enriched diet for 10 days. On Day 11, rats with hypertriglyceridemia were randomly assigned to receive oral doses of vehicle or the compound (50 mg/kg) daily for 7 consecutive days. Serum triglyceride concentrations were measured 24 hours after each administration of test agent, and blood pressure was measured 18 hours after test agent administration. FIG. 3A shows that the test compound effectively lowered serum triglyceride concentrations in these rats, and FIG. 3B shows that the rats treated with the test compound had significantly lower blood pressure than did those treated with vehicle.
Basal glucose uptake of 3T3-L1 cells was measured in the presence of two different concentrations of the test compound (0.1 nM and 0.1 μM). Cells were incubated at 37° C. for 48 hours with vehicle or the test compound, and then further incubated with 14C-deoxyglucose for an additional 30 min at 22° C. The cells were washed and lysed, and the total radioactivity in the cells was measured. FIG. 4 shows that the glucose uptake increased over the basal level in cells treated with the test compound. This result suggests that this test compound stimulates glucose uptake in differentiated adipocytes.
In an experiment studying the expression of PPAR-γ in the adipose tissue of mice, epididymal fat was collected from six different ob/ob mice either treated with vehicle or the test compound (50 mg/kg) for 10 days, homogenized in lyses buffer, and centrifuged. A total of 30 mg of protein was loaded on to SDS polyacrylamide gel, immunoblotted, and probed with anti PPAR-γ antibody raised against a 15-residue synthetic peptide containing conserved sequences of PPAR-γ (see FIG. 5A). The bands were quantified and represented in bar graphs (see FIG. 5B). The expression levels of PPAR-γ in the tissues from vehicle-treated and the compound-treated animals did not differ from each other.
The tests described above and illustrated in the figures show that the compounds according to the present invention lower blood glucose concentrations, lower serum triglyceride concentrations, lower systolic blood pressure, and increase glucose uptake by adipose tissue, but do not affect the expression of PPAR-γ by adipose tissue.

## Claims

1. A method for lowering blood glucose in a subject comprising administering to said subject an effective blood glucose lowering amount of a compound represented by formula II: 



wherein stereocenters *, if present, are R or S; 
dotted lines indicate that a double bond is absent; 
A, A′, and C are H; 
B and B′ are H; 
R′, R′′, and R′′′ are H or C1-C20 alkoxycarbonyl; 
t, x, y and u represent the integer 2; 
w represents the integer 1; and 
X═O.

2. A method for lowering serum triglyceride in a subject comprising administering to said subject an effective serum triglyceride lowering amount of a compound represented by formula II: 



wherein stereocenters *, if present, are R or S; 
dotted lines indicate that a double bond is absent; 
A, A′, and C are H; 
B and B′ are H; 
R′, R′′, and R′′′ are independently H or C1-C20 alkoxycarbonyl; 
t, x, y and u represent the integer 2; 
w represents the integer 1; and 
X═O.

3. A method for lowering blood pressure in a subject comprising administering to said subject an effective blood pressure lowering amount of a compound represented by formula II: 



wherein stereocenters *, if present, are R or S; 
dotted lines indicates that a double bond is absent; 
A, A′, and C are H; 
B and B′ are H; 
R′, R′′, and R′′′ are independently H or C1-C20 alkoxycarbonyl; 
t, x, y and u represent the integer 2; 
w represents the integer 1; and 
X═O.

4. The method of claim 1 wherein said alkoxycarbonyl is methoxycarbonyl.

5. The method of claim 2 wherein said alkoxycarbonyl is methoxycarbonyl.

6. The method of claim 3 wherein said alkoxycarbonyl is methoxycarbonyl.