WO2012054817A1

WO2012054817A1 - Delaying and/ or treating hyperglycemia, including diabetes, with poly(alkyl carboxylic acids)

Info

Publication number: WO2012054817A1
Application number: PCT/US2011/057254
Authority: WO
Inventors: Brooke Ligon
Original assignee: MITOKINE BIOSCIENCE LLC
Current assignee: MITOKINE BIOSCIENCE LLC
Priority date: 2010-10-22
Filing date: 2011-10-21
Publication date: 2012-04-26
Anticipated expiration: 2013-04-22

Abstract

The present invention is directed to a method of delaying the onset of and/or treating hyperglycemia, particularly hyperglycemia caused by diabetes, comprising administering to a mammal in need thereof an effective amount of a poly(alkyl carboxylic acid), or a pharmaceutically acceptable salt thereof. Also described is a pharmaceutical composition for the treatment of hyperglycemia, comprising an effective amount of a poly(alkyl carboxylic acid), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier. Also disclosed is a kit, comprising an effective amount of a poly(alkyl carboxylic acid), or a pharmaceutically acceptable salt thereof, and an effective amount of insulin.

Description

DELAYING AND/OR TREATING HYPERGLYCEMIA, INCLUDING DIABETES, WITH POLY(ALKYL CARBOXYLIC ACIDS)

RELATED APPLICATION

This application claims the benefit of U.S. Provisional Application No. 61/405,958, filed on October 22, 2010. The entire teachings of the aforementioned application are incorporated herein by reference.

BACKGROUND OF THE INVENTION

Hyperglycemia, or high blood sugar, can cause a variety of health complications. One common cause of hyperglycemia is diabetes mellitus, often referred to simply as "diabetes," which is a condition in which a person has a high blood level of the sugar glucose. This abnormally high level of glucose is generally either because the person's pancreas does not produce sufficient insulin or because other cells in the person's body do not properly respond to insulin that is produced. Insulin is a hormone that enables body cells to absorb glucose which is used by the cells to produce energy.

Although there are many types of diabetes, the two most common types are called type 1 diabetes and type 2 diabetes. In type 1 diabetes, a person's pancreas fails to produce sufficient insulin and the person is usually required to inject insulin. Type 2 diabetes is proposed to result from insulin resistance, a condition in which cells fail to use insulin properly, sometimes combined with insulin deficiency.

Currently, there are hundreds of millions of diabetic people in the world and the incidence of diabetes is increasing at a substantial rate.

Because of this, there has been and currently is an extremely large amount of research directed to better understand the causes of diabetes and to develop new and better treatments. Despite this, no currently available treatments are entirely satisfactory and there is a need for new treatments efficacious for treating diabetic mammals, and for delaying or preventing the onset of diabetes to susceptible mammals. SUMMARY OF THE INVENTION

The present invention provides a method of delaying the onset of and/or treating hyperglycemia, comprising periodically administering to a mammal in need thereof an effective amount of a poly(alkyl carboxylic acid), or a pharmaceutically acceptable salt thereof.

Another embodiment of the present invention is a method of stimulating pancreatic islet cell growth, comprising administering to a mammal in need thereof an effective amount of a poly(alkyl carboxylic acid), or a pharmaceutically acceptable salt thereof.

Another embodiment of the present invention is a method for regulating body mass and/or maintaining lean body mass in mammals, comprising

administering to a mammal in need thereof an effective amount of insulin and an effective amount of a poly(alkyl carboxylic acid), or a pharmaceutically acceptable salt thereof.

In another embodiment, the method relates to maintaining glucose homeostasis, comprising administering to a mammal in need thereof an effective amount of a poly(alkyl carboxylic acid), or a pharmaceutically acceptable salt thereof. In another embodiment, the method relates to decreasing exogenous insulin required to maintain glucose homeostasis, comprising periodically administering to a mammal in need thereof an effective amount of a poly(alkyl carboxylic acid), or a pharmaceutically acceptable salt thereof.

The present invention also provides a method of delaying the onset of and/or treating diabetes meilitus, comprising administering to a mammal in need thereof an effective amount of a poly(alkyl carboxylic acid), or a pharmaceutically acceptable salt thereof.

The current invention also provides a method of delaying the onset of and/or treating diabetes meilitus, comprising administering to a mammal in need thereof an effective amount of insulin and an effective amount of a poly(alkyl carboxylic acid), or a pharmaceutically acceptable salt thereof.

Another embodiment of the present invention is a pharmaceutical composition for the treatment of hyperglycemia, comprising an effective amount of a poly(alkyl carboxylic acid), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

Yet another embodiment of the present invention is a kit, comprising a poly(alkyl carboxylic acid), or a pharmaceutically acceptable salt thereof, and an effective amount of insulin.

The present invention provides a new and efficacious treatment of hyperglycemia, particularly hyperglycemia caused by diabetes, involving poly(alkyl carboxylic acids), which have been found efficacious for delaying the onset of and treating hyperglycemia, including diabetes. Other advantages of the invention include reducing the insulin requirements in a mammal being treated for diabetes, maintaining lean body mass in mammals with diabetes, and maintaining glucose homeostasis in a mammal using a poly(alkyl carboxylic acid), or a pharmaceutically acceptable salt thereof.

BRIEF DESCRIPTION OF THE FIGURES FIG. 1 is a plot of average fasting plasma glucose (fpg) versus time and illustrates the average fasting plasma glucose level of female NOD mice during treatment with insulin, insulin + hematoporphyrin (Hem), or insulin + Hem + GABA over a time period of 28 weeks.

FIG. 2 is a plot of mass versus time, illustrating the mass of female NOD mice without treatment (control) or upon treatment with insulin, insulin + hematoporphyrin (Hem), or insulin + Hem + GABA over a time period of 30 weeks.

FIG. 3 is a plot of average units of insulin per week versus time and illustrates the average weekly insulin requirement of diabetic, female NOD mice during treatment with insulin, insulin + hematoporphyrin (Hem), or insulin + GABA over a time period of 9 weeks.

FIG. 4 is a plot of average weekly blood glucose (bg) versus time and illustrates the average weekly blood glucose level of diabetic, female NOD mice during treatment with insulin, insulin + hematoporphyrin (Hem), or insulin + GABA over a time period of 9 weeks. DETAILED DESCRIPTION OF THE INVENTION

As used in the description of this invention, the terms set forth below have the following definitions.

As used herein, the singular forms "a," "an" and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a poly(alkyl carboxylic acid)" can include a plurality of such molecules. Further, the plurality can comprise more than one of the same molecule or a plurality of different molecules.

The term "hyperglycemia" as used herein, means high blood sugar and generally corresponds to a plasma glucose level (also referred to as blood glucose level) greater than normal as determined by diagnostic indicators. For example, a plasma glucose level greater than or equal to about 180 mg/dl or greater than or equal to about 200 mg/dl is generally considered greater than normal. One of ordinary skill in the art would readily recognize definitions, including that of hyperglycemia, are subject to change over time and the latest definitions and standards disclosed by organizations such as the World Health Organization and the American Diabetes Association can be used to define hyperglycemia as provided in the present invention.

The term "hyperglycemia" is also intended to include those individuals with chronic hyperglycemia, hyperinsulinemia, impaired glucose homeostasis or tolerance, insulin resistance, and diabetes. Plasma glucose levels in hyperglycemic individuals include, for example, glucose concentrations greater than normal as determined by reliable diagnostic indicators. Such hyperglycemic individuals are at risk or predisposed to developing overt clinical symptoms of diabetes mellitus.

As used herein, the term "diabetes" is intended to mean all diabetic conditions, including, without limitation, diabetes mellitus, genetic diabetes, type 1 diabetes, type 2 diabetes, and gestational diabetes. The term "diabetes" also refers to the chronic disease characterized by relative or absolute deficiency of insulin that results in glucose intolerance. Type 1 diabetes is also referred to as insulin dependent diabetes mellitus (IDDM) and also includes, for example, juvenile-onset diabetes mellitus. Type 1 is primarily due to the loss of pancreatic β-cells. Type 2 diabetes mellitus is also known as non-insulin dependent diabetes mellitus

(NIDDM) and is characterized, in part, by impaired insulin release following a meal. Insulin resistance can also be a factor leading to the occurrence of type 2 diabetes mellitus. Genetic diabetes is due to mutations which interfere with the function and regulation of p-cells.

Diabetes, as used herein, is characterized as a fasting level of plasma glucose greater than or equal to about 126 mg/dl or as a plasma glucose level greater than or equal to about 180 mg/dl as assessed at about 2 hours following the oral

administration of a glucose load of about 75 grams or following a meal. As understood by the skilled artisan, characteristics used in identifying diabetes are subject to change and the latest standards, such as those disclosed by the World Health Organization, can be used to define diabetes as provided in the present invention. In some embodiments of the present invention, diabetes is arbitrarily defined as a plasma glucose level of greater than or equal to about 200 mg/dl.

The term "diabetes" is also intended to include those mammals with hyperinsulinemia, impaired glucose homeostasis or tolerance, and insulin resistance.

As used herein, the term "diabetic complications" refers to medical/clinical problems that occur more often in patients diagnosed with diabetes. As

contemplated herein, diabetic complications include medical/clinical problems that stem from changes in blood vessels and/or nerves as a result of diabetes. These include, and are not limited to, skin conditions (e.g., bacterial infections, fungal infections, diabetic dermopathy), necrobiosis lipoidica, diabeticorum (e.g., bullosis diabeticorum), eruptive xanthomatosis, allergic skin reactions, digital scleroris, disseminated granuloma annulare, and acanthosis nigricans), gum disease, eye disorders (e.g., glaucoma, cataracts, retinopathy), kidney disease, neuropathy (e.g., diabetic mononeuropathy, neuropathic pain, non-ketotic hyperglycemic coma, thoracic radiculopathy, systemic neuropathy, distal systemic polyneuropathy, proximal neuropathy, femoral neuropathy, neuropathic antrhropathy, cranial neuropathy, autonomic neuropathy, compression neuropathy, and diabetic amyotrophy), gout, non-proliferative retinopathy (e.g., diabetic macular edema, vitreous hemorrhage with retinal detachment, anterior ischemic optic neuropathy, proliferative retinopathy with retinal vascular microaneurysm, neovascularization hemorrhage, and retinal detachment), iris disorders (e.g., rubeosis iridis), gastrointestinal disorders (e.g., gastroparesis, enteric neuropathy, diabetic fatty liver, diabetic steatonecrosis), impaired wound healing, and cardiovascular

diseases/disorders (e.g., hypertension, heart disease, heart attack, stroke, ischemia).

The term "mammal," as used herein, describes the subject to which the treatment with the compositions and methods of the present invention is provided. Mammalian species that benefit from the disclosed methods of treatment include, but are not limited to, apes, chimpanzees, orangutans, humans, monkeys; and domesticated animals (e.g., pets) such as dogs, cats, mice, rats, guinea pigs, and hamsters. Diabetes also occurs in other species, such as sheep, horses, cattle, and pigs. When the mammal is a human being, it is often referred to as a patient.

"Treating" or "treatment" refers to obtaining desired pharmacological and/or physiological effect. The effect can include achieving, partially or substantially, one or more of the following results: partially or totally reducing the extent of the disease, disorder, or syndrome; ameliorating or improving a clinical symptom or indicator associated with the disease, disorder, or syndrome; delaying, inhibiting or decreasing the likelihood of the progression of the disease, disorder or syndrome.

As used herein, "delaying the onset of refers to postponing the onset or development of disease, disorder, or syndrome.

As used herein, "poly(alkyl carboxylic acid)" refers to compounds, or pharmaceutically acceptable salts thereof, comprising at least two alkyl carboxylic acids and a scaffold to which the alkyl carboxylic acids are linked and which provides the molecule with efficacy for diabetes when administered to a mammal. Preferably, the poly(alkyl carboxylic acid) comprises two to ten, two to eight, two to four, two, or four alkyl carboxylic acids. Preferably, the alkyl carboxylic acid is propionic or butyric carboxylic acid. More preferably, the alkyl carboxylic acid is propionic acid. Most preferably, there are exactly two propionic acids.

The scaffold can be C1-C7 alkylene. As used herein, "alkylene" refers to an optionally substituted unsaturated aliphatic branched or straight-chain hydrocarbon radical having the specified number of carbon atoms. Preferably, the alkylene is a C1-C7, C1-C5, C1-C3 or CI alkylene. More preferably, the alkylene is substituted.

The scaffold can also be C1-C7 alkyl. As used herein, "alkyl" refers to an optionally substituted saturated hydrocarbon radical having the specified number of atoms in a linear or branched arrangement. Preferably, the alkyl is a C1-C7, C1-C5, C1-C3 or CI alkyl. More preferably, the alkyl is substituted.

Examples of optional substituents for alkylene or alkyl include saturated or unsaturated carbocyclic or heterocyclic groups, aromatic or heteroaromatic groups, =0, =S, =NH, and those listed below for a substitutable carbon of an aryl or heteroaryl group.

"Carbocyclic" or "carbocyclyl," as used herein, refers to an optionally substituted mono-, bi-, or other multicyclic ring having only carbon ring atoms and includes 3-12-membered, 3-8-membered, 3-6-membered, or 3-5-membered saturated, partially saturated or unsaturated aliphatic cyclic hydrocarbon rings or 6-14-membered or 6- 10-membered aryl groups.

"Heterocyclic" or "heterocyclyl," as used herein, refers to an optionally substituted mono-, bi-, or other multicyclic 3-13-membered, 3-8-membered, 3-6- membered, 3-5-membered, or 5-membered saturated or unsaturated aliphatic or aromatic ring containing 1, 2, 3, 4 or 5 heteroatoms independently selected from N, O or S. When one heteroatom is S, it can be optionally mono- or di-oxygenated

(i.e., -S(O)- or -S(0)₂-). Examples of monocyclic heterocyclyls include, but are not limited to, 2,3-dihydro-lH-pyrrole, 2,5-dihydro-lH-pyrrole, azetidine, pyrrolidine, piperidine, piperazine, azepane, hexahydropyrimidine, tetrahydrofuran,

tetrahydropyran, morpholine, and thiomorpholine.

Examples of suitable substituents for a carbocyclic or heterocyclic group include =0, =S, =NH, and those listed below for a substitutable carbon of an aryl or heteroaryl group. Preferably, the carbocyclic or heterocyclic group is substituted with halogen, alkyl, amino, alkylamino, dialkylamino, alkylene, alkoxy, hydroxyl, nitro, cyano, hydroxyalkyl, haloalkoxy, or haloalkyl, or =0. More preferably, the carbocyclic or heterocyclic group is substituted with alkyl, hydroxyalkyl, or alkylene, or =0. "Aromatic" or "aryl," as used herein, refers to an optionally substituted mono-, bi-, or other multi-carbocyclic, aromatic ring system. The aryl group can optionally be fused to one or more rings selected from aryls, cycloalkyls, and heterocyclyls. Aryls can have from six to fourteen ring members, such as from six to ten ring members. One or more hydrogen atoms may also be replaced by a substituent group. Nonlimiting examples of aryl groups include phenyl, naphthyl, biphenyl, and anthracenyl.

"Heteroaromatic" or "heteroaryl," as used herein, refers to an aromatic radical having from one to four hetero atoms or hetero groups selected from O, N, or S in a single or fused heterocyclic ring system, having from five to fifteen ring members, such as a heteroaryl ring system having from five to ten ring members. When one heteroatom is S, it can be optionally mono- or di-oxygenated (i.e., -S(O)- or -S(0)₂-). One or more hydrogen atoms may also be replaced by a substituent group. Nonlimiting examples of heteroaryl groups include furanyl, indolyl, pyridinyl, thiazolyl, thiadiazoyl, isoquinolinyl, pyrazolyl, oxazolyl, oxadiazolyl, triazolyl, and pyrrolyl groups.

Examples of substituents for an aryl or heteroaryl group include halogen, alkyl, amino, alkylamino, dialkylamino, alkylene, alkoxy, hydroxyl, nitro, cyano, hydroxyalkyl, haloalkoxy, haloalkyl, thio, thioalkyl, thioalkoxy, aryl, heteroaryl, carbocyclyl, and heterocyclyl. Preferably, the aryl or heteroaryl group is substituted with halogen, alkyl, amino, alkylamino, dialkylamino, alkylene, alkoxy, hydroxyl, nitro, cyano, hydroxyalkyl, haloalkoxy, or haloalkyl. More preferably, the aryl or heteroaryl group is substituted with alkyl, hydroxyalkyl, or alkylene.

In one embodiment, the scaffold comprises an alkyl or alkylene group alternating with a carbocyclic or heterocyclic group. Preferably, the scaffold comprises a (Cl)alkyl or (Cl)alkylene group alternating with a carbocyclic or heterocyclic group. Alternatively, the scaffold comprises an alkyl or alkylene group alternating with a heterocyclic group. Preferably, the scaffold comprises a (Cl)alkyl or (Cl)alkylene group alternating with a heterocyclic group.

In one embodiment, the scaffold comprises a conjugated system comprising alternating alkylene and heterocyclyl groups. Preferably, the scaffold comprises a conjugated system comprising alternating alkylene and heteroaromatic groups. More preferably, the scaffold comprises comprises a conjugated system comprising alternating (Cl)alkylene and heteroaromatic groups.

In some embodiments, the alkylene groups and the heterocyclyl groups can be linked so as to form a cyclic structure as, for example, in hematoporphyrin.

"Cyclic," as used herein, refers to the overall structure of the scaffold, rather than its individual components (e.g. , heterocyclyls, alkyl groups, alkylene groups). A scaffold is cyclic if its individual components are connected so as to form a ring.

Alternatively, the alkylene groups and the heterocyclyl groups can be linked so as to form a linear structure as, for example, in phycocyanobilin and biliverdin. "Linear," as used herein, refers to the overall structure of the scaffold, rather than its individual components (e.g., heterocyclyls, alkyl groups, alkylene groups), and means acyclic. A scaffold is linear if its individual components are connected in such a way that they do not form a ring. It should be understood that a scaffold can be linear even if it contains individual components that are cyclic (e.g.,

heterocyclyls).

The scaffold can comprise one, two, or many saturated or unsaturated carbocyclic or heterocyclic groups. Preferably, the scaffold comprises two, three, or four saturated, partially saturated, or unsaturated carbocyclic or heterocyclic groups. Preferably, the scaffold comprises four saturated, partially saturated, or unsaturated carbocyclic or heterocyclic groups.

In a preferred embodiment, the scaffold comprises three, four or five (Cl)alkyl or (Cl)alkylene groups alternating with four heterocyclyl groups. In a more preferred embodiment, the scaffold comprises four (Cl)alkyl or (Cl)alkylene groups alternating with four heterocyclyl groups linked so as to form a cyclic structure. In another embodiment, the scaffold comprises three, four, or five (Cl)alkyl or (Cl)alkylene groups alternating with four heterocyclyl groups linked so as to form a linear structure.

In some embodiments, the heterocyclic group is a 5-membered heterocyclyl containing one, two, three, or four, and preferably one, nitrogen atom. Preferably, the heterocyclyl is a saturated or partially unsaturated heterocyclyl. The saturated or partially unsaturated heterocyclyl is optionally substituted with an oxo group.

Preferably, the saturated or partially unsaturated heterocyclyl is pyrrolidine, pyrroline, pyrrolidinone, or a partially unsaturated pyrrolidinone.

Alternatively, the heterocyclic group is a heteroaryl. Preferably, the heteroaryl is pyrrole, imidazole, pyrazole, triazole, or tetrazole. More preferably, the heteroaryl is pyrrole.

In some embodiments, the scaffold orients two alkyl carboxylic acids such that they have approximately the same spatial relationship to one another in vivo as the two propionic acids present in hematoporphyrin.

In another embodiment of the poly(alkyl carboxylic acids) of the present invention, the scaffold comprises a site (e.g., a polyunsaturated carbonyl) for covalent modification of the poly(alkyl carboxylic acid) by a receptor, particularly by an appropriately situated amino acid residue of the receptor, such as a cysteine residue.

While not wishing to be bound by this theory, we believe that the poly(alkyl carboxylic acid) may work by operating as a gamma-aminobutyric acid (GABA) receptor agonist. It is also possible that the poly(alkyl carboxylic acid) is an allosteric modulator of the insulin receptor and/or an allosteric modulator of the GABA receptor.

In a particularly preferred case, the poly(alkyl carboxylic acid) comprises porphyrin. Most preferably, the poly(alkyl carboxylic acid) is hematoporphyrin.

In yet another case, the poly(alkyl carboxylic acid) comprises a naturally occurring intermediate in the heme biosynthetic pathway, or analogs thereof.

Preferably, the poly(alkyl carboxylic acid) is selected from the group consisting of porphobilinogen, hydroxymethylbilane, uroporphyrinogen III, coproporphyrinogen III, protoporphyrinogen III, protoporphyrin IX, and heme.

In yet still another case, the poly(alkyl carboxylic acid) comprises a naturally occurring intermediate in the degradation of heme, or analogs thereof. Such intermediates include, but are not limited to bilins and/or other components of blood likely present at injury that stimulate beta cell proliferation. Preferably, the poly(alkyl carboxylic acid) is selected from the group consisting of biliverdin, bilirubin, urobilinogen, and stercobilinogen.

In yet still another case, the poly(alkyl carboxylic acid) comprises phycobilins as well as an intermediate in the biosynthesis and/or biodegradation of the phycobilins, or analogs thereof. Preferably, the poly(alkyl carboxylic acid) is selected from the group consisting of phycoerythrobilin, phycocyanobilin, phycourobiiin, and phycoviolobilin.

In one embodiment, the method comprises administering the poly(alkyl carboxylic acid) at or near a time of sustained endogenous insulin secretion, for example periprandially. The term "endogenous insulin secretion" as used herein, means secretion of insulin by pancreatic beta cells. As used herein, "periprandial" refers to a period of time beginning shortly before and ending shortly after the ingestion of a meal or snack. Many subjects or patients suffering from Type 2 diabetes are insulin-resistant and therefore experience prolonged elevation of insulin levels. In some embodiments of the present invention, the term "sustained endogenous insulin secretion" refers to the elevated levels of insulin present in type

2 diabetics.

In another embodiment, the method comprises administering the poly(alkyl carboxylic acid) at or near a time of an exogenous insulin administration. The term "at or near a time" as used herein, means the poly(alkyl carboxylic acid) is administered just before, during, or just after sustained endogenous insulin secretion or exogenous insulin administration. The term "exogenous insulin administration" as used herein, means receiving insulin from other than the subject's pancreatic beta cells, for example, from an injection of insulin.

Administration of the composition can occur at various times during the day.

In certain embodiments, the composition is administered at or near exogenous insulin administration. In one embodiment of the present invention, the composition is administered less than about 60 minutes or less than about 30 minutes or less than about to 20 minutes prior to or after administering exogenous insulin. In still other embodiments, the composition and exogenous insulin are administered at the same time. The administration of the composition and exogenous insulin can require varying dosages for a given mammal and could result in dose-to-dose variability.

"Effective amount" means that amount of active compound that elicits the desired biological response in a subject. The typical daily dose of the active substance varies within a wide range and will depend on various factors such as for example the individual requirement of each subject or patient and the route of administration. The term "mmol/kg," as used herein means "mmol" of alkyl carboxylic acid moieties of poly(alkyl carboxylic acid) per "kg" of body weight. In certain embodiments, the therapeutically effective amount of the poly(alkyl carboxylic acid) comprises approximately 0.2 mmol to approximately 0.9 mmol poly(alkyl carboxylic acid) per day per kg body weight. In one embodiment, the therapeutically effective amount of poly(alkyl carboxylic acid) comprises approximately 0.25 mmol/kg to approximately 0.6 mmol/kg. In certain

embodiments, the therapeutically effective amount of poly(alkyl carboxylic acid) comprises approximately 0.6 mmol/kg. In certain embodiments, the therapeutically effective amount of poly(alkyl carboxylic acid) comprises approximately 0.3 mmol/kg.

In embodiments in which the poly(alkyl carboxylic acid) is

hematoporphyrin, the therapeutically effective amount comprises approximately 20 to approximately 520 mg per day per kg body weight. In one embodiment, the therapeutically effective amount of hematoporphyrin comprises approximately 45- 350 mg/kg. In another embodiment, the therapeutically effective amount of hematoporphyrin comprises approximately 90-260 mg/kg. In certain embodiments, the therapeutically effective amount of hematoporphyrin comprises approximately 20 mg/kg. In certain embodiments, the therapeutically effective amount of hematoporphyrin comprises approximately 30 mg/kg. In certain embodiments, the therapeutically effective amount of hematoporphyrin comprises approximately 45 mg/kg. In certain embodiments, the therapeutically effective amount of

hematoporphyrin comprises approximately 260 mg/kg. In certain embodiments, the therapeutically effective amount of hematoporphyrin comprises approximately 350 mg/kg. In certain embodiments, the therapeutically effective amount of

hematoporphyrin comprises approximately 520 mg/kg.

In one embodiment the mammal is a human. In another embodiment, the mammal is a non-human primate. In another embodiment, the mammal is a canine. In another embodiment, the mammal is a feline. In another embodiment, the mammal is a companion animal or livestock.

As used herein, "periodic" or "periodically" refers to the repeated

administration of the compounds and/or compositions of the present invention at regular or irregular intervals of time. In one embodiment, the method comprises administering the pharmaceutical composition once, at least once, twice, at least twice, three times, at least three times, four times, or at least four times per day.

Another embodiment of the present invention is a pharmaceutical composition for the treatment of hyperglycemia, including diabetes, comprising an effective amount of the poly(alkyl carboxylic acid), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier. For use in medicines, the salts of the compounds of the invention refer to non-toxic, pharmaceutically acceptable salts.

The pharmaceutically acceptable salts of the disclosed poly(alkyl carboxylic acids) include acid addition salts and base addition salts. Any of the poly(alkyl carboxylic acids) of the present invention can be in the form of pharmaceutically acceptable salts. The term "pharmaceutically acceptable salts" embraces salts commonly used to form alkali metal salts and to form addition salts of free acids or free bases. The nature of the salt is not critical, provided that it is pharmaceutically acceptable.

Suitable pharmaceutically acceptable acid addition salts of the disclosed poly(alkyl carboxylic acids) can be prepared from an inorganic acid or an organic acid. Examples of such inorganic acids are hydrochloric, hydrobromic, hydroiodic, nitric, carbonic, sulfuric and phosphoric acid. Appropriate organic acids can be selected from aliphatic, cycloaliphatic, aromatic, arylaliphatic, heterocyclic, carboxylic and sulfonic classes of organic acids, examples of which are formic, acetic, propionic, succinic, glycolic, gluconic, maleic, embonic (pamoic), methanesulfonic, ethanesulfonic, 2-hydroxyethanesulfonic, pantothenic, benzenesulfonic, toluenesulfonic, sulfanilic, mesylic, cyclohexylaminosulfonic, stearic, algenic, β-hydroxybutyric, malonic, galactic, and galacturonic acid.

Pharmaceutically acceptable acidic/anionic salts also include, acetate,

benzenesulfonate, benzoate, bicarbonate, bitartrate, bromide, calcium edetate, camsylate, carbonate, chloride, citrate, dihydrochloride, edetate, edisylate, estolate, esylate, fumarate, glyceptate, gluconate, glutamate, glycollylarsanilate,

hexylresorcinate, hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isethionate, lactate, lactobionate, malate, maleate, malonate, mandelate, mesylate, methylsulfate, mucate, napsylate, nitrate, pamoate, pantothenate,

phosphate/diphospate, polygalacturonate, salicylate, stearate, subacetate, succinate, sulfate, hydro gensulfate, tannate, tartrate, teoclate, tosylate, and triethiodide salts.

Suitable pharmaceutically acceptable base addition salts of the disclosed poly(alkyl carboxylic acids) include, but are not limited to, metallic salts made from aluminum, calcium, lithium, magnesium, potassium, sodium and zinc or organic salts made from N,N'-dibenzylethylene-diamine, chloroprocaine, choline, diethanolamine, ethylenediamine, N-methylglucamine, lysine, arginine and procaine. All of these salts can be prepared by conventional means from the corresponding compound represented by the disclosed compound by treating, for example, the disclosed poly(alkyl carboxylic acids) with the appropriate acid or base. Pharmaceutically acceptable basic/cationic salts also include, diethanolamine, ammonium, ethanolamine, piperazine and triethanolamine salts.

In an embodiment, the pharmaceutically acceptable salt comprises a monovalent cation or a divalent cation. In a particular embodiment, the

pharmaceutically acceptable salt is a lysine salt.

In another embodiment, the monovalent cation is a monovalent metal cation and the divalent cation is a divalent metal cation. In a particular embodiment, the monovalent metal cation is a sodium cation.

The pharmaceutical compositions disclosed herein are prepared in accordance with standard procedures and are administered at dosages that are selected to reduce, prevent, eliminate, or to slow or halt the progression of, the condition being treated (see, e.g. , Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, PA, and Goodman and Gilman's The Pharmaceutical Basis of Therapeutics, McGraw-Hill, New York, N.Y., the relevant contents of which are incorporated herein by reference, for a general description of the methods for administering various agents for human therapy). The compositions of a compound represented by the disclosed poly(alkyl carboxylic acids) can be delivered using controlled or sustained-release delivery systems (e.g., capsules, biodegradable matrices). Exemplary delayed-release delivery systems for drug delivery that would be suitable for administration of the compositions of the disclosed compounds are described in U.S. Patent Nos. 5,990,092 (issued to Walsh); 5,039,660 (issued to Leonard); 4,452,775 (issued to Kent); and 3,854,480 (issued to Zaffaroni), the relevant teachings of which are incorporated herein by reference.

For preparing pharmaceutical compositions from the poly(alkyl carboxylic acids) of the present invention, pharmaceutically acceptable carriers can either be solid or liquid. Solid form preparations include powders, tablets, pills, capsules, cachets, suppositories, and dispersible granules. For example, the poly(alkyl carboxylic acids) of the present invention can be in powder form for reconstitution at the time of delivery. A solid carrier can be one or more substances that can also act as diluents, flavoring agents, solubilizers, lubricants, suspending agents, binders, preservatives, tablet disintegrating agents, or an encapsulating material. In powders, the carrier is a finely divided solid that is in a mixture with the finely divided active ingredient.

In tablets, the active poly(alkyl carboxylic acid) ingredient is mixed with the carrier having the necessary binding properties in suitable proportions and compacted in the shape and size desired.

Suitable carriers are magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium caboxymethylcellulose, a low-melting wax, cocoa butter, and the like. Tablets, powders, cachets, lozenges, fast-melt strips, capsules and pills can be used as solid dosage forms containing the active ingredient suitable for oral administration. Liquid form preparations include solutions, suspensions, and emulsions, for example, water or water propylene glycol solutions or vegetable oil emulsions. For parenteral injection, liquid preparations can be formulated in solution in aqueous polyethylene glycol solution.

Aqueous solutions suitable for oral administration can be prepared by dissolving the active poly(alkyl carboxylic acid) ingredient in water and adding suitable colorants, flavors, stabilizing agents, and thickening agents as desired.

Aqueous suspensions for oral administration can be prepared by dispersing a finely divided poly(alkyl carboxylic acid) active ingredient in water with viscous material, such as natural or synthetic gums, resins, methylcellulose, sodium

carboxymethylcellulose, and other well-known suspending agents.

The pharmaceutical composition can be in unit dosage form. In such form, the composition is subdivided into unit doses containing appropriate quantities of the active ingredient. The unit dosage form can be a packaged preparation, the package containing discrete quantities of, for example, tablets, powders, and capsules in vials or ampoules. Also, the unit dosage form can be a tablet, cachet, capsule, or lozenge itself, of it can be the appropriate amount of the any of these in packaged form. In some embodiment, the unit dosage form can be part of a kit. The dosages may be varied depending upon the requirements of the patient, the severity of the condition being treated, the compound and the route of administration being employed. Determination of the proper dosage for a particular situation is within the skill of the art. Also, the pharmaceutical composition or kit comprising the pharmaceutical composition may contain, if desired, other compatible therapeutic agents.

In general, the methods for delivering the disclosed poly(alkyl carboxylic acids) and pharmaceutical compositions of the invention in vivo utilize art- recognized protocols for delivering the agent with the only substantial procedural modification being the substitution of a poly(alkyl carboxylic acid), or a

pharmaceutically acceptable salt thereof, for the drug(s) in the art-recognized protocols. The poly(alkyl carboxylic acids) of the present invention can be administered by many routes, preferably in the form of a pharmaceutical composition adapted to such a route, and would be dependent on the condition being treated. The compounds and compositions can, for example, be administered intravascularly, orally, subcutaneously, intradermally, intramucosally, intramuscularly, or topically. It will be obvious to those skilled in the art that the following dosage forms can comprise as the active ingredient, either compounds or a corresponding

pharmaceutically acceptable salt of a compound of the present invention. A preferred method of administration for the compounds of the invention is oral administration.

For oral administration, the pharmaceutical compositions can be in the form of, for example, a tablet, capsule, suspension or liquid. The composition is preferably made in the form of a dosage unit containing a therapeutically effective amount of the active ingredient. Examples of such dosage units are tablets and capsules. For therapeutic purposes, the tablets and capsules can contain, in addition to the active ingredient, conventional carriers such as binding agents, for example, acacia gum, gelatin, polyvinylpyrrolidone, sorbitol, or tragacanth; fillers, for example, calcium phosphate, glycine, lactose, maize-starch, sorbitol, or sucrose; lubricants, for example, magnesium stearate, polyethylene glycol, silica, or talc; disintegrants, for example potato starch, flavoring or coloring agents, or acceptable wetting agents. Oral liquid preparations generally in the form of aqueous or oily solutions, suspensions, emulsions, syrups or elixirs can contain conventional additives such as suspending agents, emulsifying agents, non-aqueous agents, preservatives, coloring agents and flavoring agents. Examples of additives for liquid preparations include acacia, almond oil, ethyl alcohol, fractionated coconut oil, gelatin, glucose syrup, glycerin, hydrogenated edible fats, lecithin, methyl cellulose, methyl or propyl para-hydroxybenzoate, propylene glycol, sorbitol, or sorbic acid.

For topical administration, the pharmaceutical compositions can be in the form of, for example, an ointment, paste, cream, lotion, gel, powder, solution, spray, or patch. For therapeutic purposes, the compositions can contain a pharmaceutically acceptable carrier, a preservative, an absorption enhancer and/or a buffer. A therapeutically effective amount of a poly(alkyl carboxylic acid) of the invention can be administered alone, or in combination with one or more other therapeutic agents. Suitable therapeutic agents that are useful for treating hyperglycemia, including hyperglycemia caused by diabetes, which can be administered in combination with a compound of the invention, include, but are not limited to sulfonylureas, biguanides, thiazolidinediones, a-glucosidase inhibitors, meglitinides, dipeptidyl peptidase IV (DPP-IV) inhibitors, glucagon- like peptide- 1 (GLP-1) and GLP-1 analogs. Suitable therapeutic agents that are useful for treating complications caused by diabetes include, but is not limited to, calcium channel blockers, beta blockers, nitroglycerin, aspirin, anti-inflammatory agents, natriuretic factors, vasodilators, thrombolytic and antithrombolic agents. Other example of therapeutic agents that can be administered in combination with a compound of the present invention include, but are not limited to, insulin, GABAB receptor allosteric modulators (e.g., N,N'-dicylcopentyl-2-methylsulfanyl-5-nitropyrimidine-4,6- diamine and analogs, 2,6-di-tert-butyl-4-(3-hydroxy-2,2-dimethylpropyl)-phenol and analogs, 5,7-bis(l , 1 -dimethylethyl)-3-hydroxy-3(trifluoromethyl)-2(3H)- benzofuranone, N-[(lR,2R,4S)-bicyclo[2.2.1]hept-2-yl]-2-methyl-5-[4- (trifluoromethyl)phenyl]-4-pyrimidinamine, 2,6-di-tert-butyl-4-(3-hydroxy-2- spiropentylpropyl)-phenol), and GABAA receptor allosteric modulators (e.g., diazepam derivatives, Valium, ativan, triazolopyridazine derivatives, pyrazolo- pyridine derivatives, nicotinic carboxamide compounds, neuroactive steroids, such as androstane and pregnane derivatives, triazolophthalazine derivatives, tricyclic pyrazolo-pyridazinone analogs, barbiturates and fenamates).

Thus, the poly(alkyl carboxylic acids) of the invention can be administered as part of a combination therapy (e.g., with each other, or with one or more other therapeutic agents such as GABA and/or insulin). The compound of the invention can be administered before, after or concurrently with one or more other therapeutic agents. In some embodiments, a compound of the invention and other therapeutic agent can be co-administered simultaneously (e.g., concurrently) as either separate formulations or as a joint formulation. Alternatively, the agents can be administered sequentially, as separate compositions, within an appropriate time frame, as determined by the skilled clinician (e.g., a time sufficient to allow an overlap of the pharmaceutical effects of the therapies); for example, as part of a kit. A compound of the invention and one or more other therapeutic agents can be administered in a single dose or in multiple doses, in an order and on a schedule suitable to achieve a desired therapeutic effect. Suitable dosages and regimens of administration can be determined by a clinician and are dependent on the agent(s) chosen, pharmaceutical formulation and route of administration, various patient factors and other

considerations.

Typically, the efficacy of a pharmacological agent is directly related to its plasma concentration, wherein efficacy is reduced as plasma concentration falls. Thus, in standard multi-dosing regimens, a pharmacological agent is administered on a dosage schedule that is designed to maintain a pre-determined or optimal plasma concentration in the subject undergoing treatment. When the agent is administered at dosage intervals that are longer than the optimal interval(s), its plasma concentration can fall to undesirably low levels before the next dose is administered, with a concomitant decrease in efficacy.

EXEMPLIFICATION

A description of example embodiments of the invention follows.

Example 1 : Prevention/Delay Treatments. Treatment paradigms to prevent or delay diabetes onset were tested in female NOD mice (Jackson Labs) beginning at 8 weeks of age. Normally, 90% of female NOD mice are diabetic by 30 weeks of age. To monitor glucose control, the mice were allowed access to food ad lib overnight. A 12-hour light-dark cycle was maintained and mice were housed in accordance with the NIH Animal Care and Use guidelines. Food was removed for eight hours during the day and the fasting plasma glucose was tested daily prior to and at the end of the eight-hour fast. At the end of the eight-hour fast, the mice were treated with one of three treatment paradigms, then immediately allowed access to food. The three treatment paradigms to assess prevention or delay of diabetes onset were as follows: one group received 0.4 units of insulin subcutaneously once per day; the second group received 0.4 units insulin and 30 μg/g GABA and 180 μg/g hematoporphyrin (Hem) once per day; and the third group received 0.4 units insulin and 180 μg/g Hem once per day.

Mice that had a plasma glucose >200 mg/dl for three consecutive fasting measurements (which equates to three consecutive days) were considered diabetic and subcutaneous insulin injections were increased to twice a day. The insulin dose was adjusted daily to keep the average fasting plasma glucose at or below 200 mg/dl. Table 1 shows the average plasma glucose (fbg) of each experimental group for each week between 13 and 32 weeks of age. At 32 weeks, n = 4 for the insulin + Hem group, n = 3 for the insulin + GABA + Hem group, and n = 3 for the insulin only treatment group (where n is the number of mice in each treatment group).

The results demonstrate that treatment with insulin and hematoporphyrin (Hem) was the most successful prevention treatment. Only the insulin + Hem group maintained an average fbg under 200 mg/dl (FIG. 1). At 28 weeks of age, 3 out of 4 of the mice in this treatment group remained free of diabetes. In the insulin only treatment and the insulin + GABA + Hem group, only one out of three of the mice in the group remained free of diabetes at 32 weeks of age.

Table 1. Average fasting plasma glucose (mg/dl) of NOD mice aged 13 to 32 weeks as a function of treatment paradigm.

13 15 17 19 21 22 24 26 28 30 32

Treatment week week week week week week week week week week week Paradigm s s s s s s s s s s s

Insulin + Hem 113 159 175 174 168 159 167 147 180 176 177

Insulin +

92 101 103 103 254 292 267 248 298 295 292 GABA + Hem

Insulin only 82 114 162 134 150 188 195 260 277 255 253 Example 2: Hematoporphyrin and Weight. Oral treatment with

hematoporphyrin had a positive effect on maintaining lean body mass of NOD mice, Mice were weighed every two weeks while on the prevention treatments described in Example 1. As in Example 1 , the mice were divided into treatment groups in an effort to prevent or delay diabetes onset. The control group received vehicle (30 μΐ vegetable oil) to the fur, which was consumed orally by grooming. The insulin only treatment group received 0.4 units insulin subcutaneously once per day at the end of the fast, just prior to feeding. The insulin + Hem group received 0.4 units insulin subcutaneously and 180 μg/g Hem diluted in 30 μΐ vegetable oil and administered to the fur, which each mouse consumed orally by grooming just prior to being fed. The final group received 0.4 units insulin subcutaneously, 180 μg/g Hem diluted in 30 μΐ vegetable oil, and 30 μg/g GABA as a 50 mg/ml solution in water and administered orally by pipette. FIG. 2 is a plot of the average weight of three or four mice (as indicated) in the specified treatment group from 12 weeks of age to 32 weeks of age. Both groups receiving oral hematoporphyrin were approximately greater than or equal to 14% leaner than NOD mice receiving insulin treatment alone and approximately greater than or equal to 10% leaner than the control NOD mice receiving vehicle.

Example 3: Treatment/Reversal Experiments. Diabetic female NOD mice (NOD/ShiLtJ, Taconic) were treated with GABA or hematoporphyrin in conjunction with insulin to reduce their insulin-dependence. The mice were housed in accordance with NIH Animal Use and Welfare guidelines with a 12-hour light-dark cycle beginning at 9 weeks of age. The animals were fed a standard rodent diet ad lib. Blood glucose was measured weekly using a Freestyle glucometer, and then twice daily once they became diabetic.

Mice were considered diabetic if one non-fasting blood glucose measurement was >200 mg/dl. For eight weeks before the mice became diabetic, their average fasting blood glucose was 102 mg/dl. Once diabetic, blood glucose levels were measured twice daily, approximately eight hours apart. All of the mice received insulin subcutaneously twice daily and the dosage was adjusted as needed.

Three diabetic mice were randomly designated as controls and were given the amino acid lysine (8 mg/kg) orally (PO) twice a day before the administration of insulin. Eight diabetic mice were treated with 60 or 90 mg/kg GABA in lieu of lysine, but were otherwise treated identically to the control group. Three diabetic mice were treated with 180 mg/kg hematoporphyrin diluted in water and

administered orally in two 30-μ1 boluses, but were otherwise treated identically to the control group.

FIG. 3 illustrates the average weekly insulin requirement of diabetic, female NOD mice during treatment with insulin + lysine, insulin + hematoporphyrin (Hem), or insulin + GAB A over a time period of 9 weeks. In the diabetic mice treated only with insulin and lysine, the required insulin dose ranged from 0.030-0.047 units/day over a 9-week study period. In the diabetic mice treated with GABA, the required insulin dose ranged from 0.004 to 0.020 units/day over the 9-week study period. In the diabetic mice treated with hematoporphyrin, the required insulin dose ranged from 0.005 to 0.025 units/day over the 9-week study period.

FIG. 4 illustrates the average weekly blood glucose level of diabetic, female NOD mice during treatment with insulin + lysine, insulin + hematoporphyrin (Hem), or insulin + GABA over a time period of 9 weeks. During the 9-week study period, the blood glucose of the control mice ranged from 362 to 463 mg/dl. The blood glucose of the GABA-treated mice ranged from 261 to 352 mg/dl and the blood glucose of the hematoporphyrin-treated mice ranged from 98 to 470 mg/dl.

In the GABA-treated group, 3 out of 8 mice became insulin-independent, no longer requiring daily insulin. In the hematoporphyrin-treated group, 1 out of 3 mice became insulin-independent.

The relevant teachings of all patents, published applications and references cited herein are incorporated by reference in their entirety.

While this invention has been particularly shown and described with references to example embodiments thereof, it will be understood by those skilled in the art that various changes in form and details can be made therein without departing from the scope of the invention encompassed by the appended claims.

Claims

What is claimed is:

1. A method of delaying the onset of and/or treating hyperglycemia in a

mammal in need thereof, comprising periodically administering to the mammal an effective amount of a poly(alkyl carboxylic acid), or a pharmaceutically acceptable salt thereof.

2. A method for stimulating islet cell growth in a mammal in need thereof, comprising periodically administering to the mammal an effective amount of a poly(alkyl carboxylic acid), or a pharmaceutically acceptable salt thereof.

3. A method for regulating body mass or maintaining lean body mass of a

mammal, comprising periodically administering to the mammal in need thereof an effective amount of insulin and an effective amount of a poly(alkyl carboxylic acid), or a pharmaceutically acceptable salt thereof.

4. A method of delaying the onset of and/or treating diabetes mellitus in a mammal in need thereof, comprising periodically administering to the mammal an effective amount of a poly(alkyl carboxylic acid), or a pharmaceutically acceptable salt thereof.

5. A method of delaying the onset of and/or treating diabetes mellitus in a mammal in need thereof, comprising periodically administering to the mammal an effective amount of insulin and an effective amount of a poly(alkyl carboxylic acid), or a pharmaceutically acceptable salt thereof.

6. The method of any one of Claims 1-5, wherein the poly(alkyl carboxylic acid) or pharmaceutically acceptable salt thereof comprises porphyrin.

1 The method of Claim 6, wherein the poly(alkyl carboxylic acid) is hematoporphyrin, or a pharmaceutically acceptable salt thereof.

The method of any one of Claims 1-5, wherein the poly(alkyl carboxylic acid) or pharmaceutically acceptable salt thereof contains two alkyl carboxylic acid moieties.

The method of Claim 8, wherein each alkyl carboxylic acid is propionic acid.

The method of any one of Claims 1-5, 8, or 9, wherein the poly(alkyl carboxylic acid) includes a scaffold comprising (Cl)alkyl or (Cl)alkylene groups alternating with heterocyclyl groups.

The method of Claim 10, wherein the scaffold comprises four (Cl)alkyl or (Cl)alkylene groups alternating with four heterocyclyl groups linked so as to form a cyclic structure.

The method of Claims 10 or 11 , wherein the scaffold comprises three, four, or five (Cl)alkyl or (Cl)alkylene groups alternating with four heterocyclyl groups linked so as to form a linear structure.

The method of any one of Claims 10-12, wherein the heterocyclyl is a 5- membered heterocyclyl containing one nitrogen atom optionally substituted with an oxo group.

The method of any one of Claims 10-12, wherein the heterocyclic group is: a heteroaryl group selected from the group consisting of pyrrole, imidazole, pyrazole, triazole, and tetrazole; or

a saturated or partially unsaturated heterocyclyl selected from the group consisting of pyrrolidine, pyrroline, pyrrolidinone, and a partially unsaturated pyrrolidinone. The method of any one of Claims 1-14, wherein the poly(alkyl carboxylic acid) or pharmaceutically acceptable salt thereof is administered orally.

The method of any one of Claims 1-15, wherein the effective amount of the poly(alkyl carboxylic acid) or pharmaceutically acceptable salt thereof is about 0.2 mmol/kg to about 0.9 mmol/kg.

The method of Claim 1, wherein the hyperglycemia is caused by diabetes mellitus.

The method of Claim 3 or Claim 5, wherein the poly(alkyl carboxylic acid) or pharmaceutically acceptable salt thereof is administered at or near the time of exogenous insulin administration or sustained endogenous insulin secretion.

The method of any one of Claims 1-5, wherein the poly(alkyl carboxylic acid) is selected from the group consisting of biliverdin, bilirubin, urobilinogen, and stercobilinogen, or a combination thereof.

The method of any one of Claims 1-5, wherein the poly(alkyl carboxylic acid) is selected from the group consisting of porphobilinogen,

hydroxymethylbilane, uroporphyrinogen III, coproporphyrinogen III, protoporphyrinogen III, protoporphyrin IX, and heme, or a combination thereof.

The method of any one of Claims 1-5, wherein the poly(alkyl carboxylic acid) is selected from the group consisting of phycoerythrobilin,

phycocyanobilin, phycourobilin, and phycoviolobilin, or a combination thereof.

22. The method of any one of Claims 1-21, wherein the mammal is a human.

23. The method of any one of Claims 1 , 2 and 4, further comprising periodically administering to the mammal an effective amount of insulin.

24. A pharmaceutical composition for the treatment of hyperglycemia,

comprising an effective amount of a poly(alkyl carboxylic acid), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

25. A kit, comprising a poly(alkyl carboxylic acid), or a pharmaceutically

acceptable salt thereof, and insulin.