阅读说明：本技术 增强二甲双胍作用的化合物 (Compounds for enhancing metformin action ) 是由 D·M·巴伦 N·博努尔 Y·拉蒂诺 坂本圭 M·桑德斯 于 2019-09-25 设计创作，主要内容包括：本发明整体涉及包含以下项的药物：二甲双胍或其盐；以及具有通式(I)的化合物或其盐。本发明还涉及包含用于预防或治疗代谢疾病或心血管疾病的药物的药物组合物。(The present invention generally relates to a medicament comprising: metformin or a salt thereof; and a compound having the general formula (I) or a salt thereof. The present invention also relates to a pharmaceutical composition comprising a medicament for preventing or treating metabolic diseases or cardiovascular diseases.)

1. A medicament, the medicament comprising:

(i) metformin; and

(ii) compounds having the general formula (I)

Wherein, in compound (ii), R1, R2, R3, R4, R5, R6, R7, and R8 are each independently: h; OH; OMe; an O-glycoside; halogen; an aldehyde; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; a nitro group; a sulfonate ester; a sulfate ester; optionally substituted and/or optionally branched C1 to C20 alkyl; c2 to C20 alkenyl optionally substituted and/or optionally branched; a C4 to C20 polyalkenyl group optionally substituted and/or optionally branched; c2 to C20 alkynyl optionally substituted and/or optionally branched; or a C4 to C20 polyalkynyl group which is optionally substituted and/or optionally branched.

2. The medicament of claim 1, wherein in compound (ii), R1, R2, R3, R4, R7 and R8 are each independently: h; OH; OMe; an O-glycoside; halogen; an aldehyde; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; a nitro group; a sulfonate ester; a sulfate ester; optionally substituted and/or optionally branched C1 to C20 alkyl; c2 to C20 alkenyl optionally substituted and/or optionally branched; a C4 to C20 polyalkenyl group optionally substituted and/or optionally branched; c2 to C20 alkynyl optionally substituted and/or optionally branched; or C4 to C20 polyalkynyl optionally substituted and/or optionally branched; r5 is: h; OMe; an O-glycoside; halogen; an aldehyde; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; a nitro group; a sulfonate ester; a sulfate ester; optionally substituted and/or optionally branched C1 to C20 alkyl; c2 to C20 alkenyl optionally substituted and/or optionally branched; a C4 to C20 polyalkenyl group optionally substituted and/or optionally branched; c2 to C20 alkynyl optionally substituted and/or optionally branched; or C4 to C20 polyalkynyl optionally substituted and/or optionally branched; r6 is: h; OH; an O-glycoside; halogen; an aldehyde; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; a nitro group; a sulfonate ester; a sulfate ester; optionally substituted and/or optionally branched C1 to C20 alkyl; c2 to C20 alkenyl optionally substituted and/or optionally branched; a C4 to C20 polyalkenyl group optionally substituted and/or optionally branched; c2 to C20 alkynyl optionally substituted and/or optionally branched; or a C4 to C20 polyalkynyl group which is optionally substituted and/or optionally branched.

3. A medicament as claimed in any preceding claim, wherein in compound (ii), R1, R2, R3, R4, R7 and R8 are each independently: h; OH; OMe; an O-glycoside; halogen; an aldehyde; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; a nitro group; a sulfonate ester; a sulfate ester; r5 is: h; OMe; an O-glycoside; halogen; an aldehyde; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; a nitro group; a sulfonate ester; a sulfate ester; r6 is: h; OH; an O-glycoside; halogen; an aldehyde; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; a nitro group; a sulfonate ester; a sulfate ester.

4. The medicament of any preceding claim, wherein in compound (ii), R1, R3 and R8 are each independently: h; OH; OMe; an O-glycoside; halogen; an aldehyde; a carboxylic acid; r2 is: OH; OMe; an O-glycoside; halogen; an aldehyde; a carboxylic acid; r4 is: h; OH; an O-glycoside; halogen; an aldehyde; a carboxylic acid; r5 is: h; OMe; an O-glycoside; halogen; an aldehyde; a carboxylic acid; r6 is: h; OH; an O-glycoside; halogen; an aldehyde; a carboxylic acid; and R7 is: h; OH; an O-glycoside; halogen; an aldehyde; a carboxylic acid.

5. The medicament of any preceding claim, wherein in compound (ii), R1, R3 and R8 are each independently: h; OH; OMe; r2 is: OH; OMe; r4 is: h; OH; r5 is: h; OMe; r6 is: h; OH; and R7 is: h; and (5) OH.

6. The medicament of any preceding claim, wherein in compound (ii), R1, R3, R6 and R8 are each independently: h; r2 is: OH; OMe; r4 is OH; r5 is: h; OMe; and R7 is: h; and (5) OH.

7. A medicament according to any preceding claim, wherein compound (ii) is lisianthrin (7-methoxy-9, 10-dihydrophenanthrene-2, 5-diol, CAS number 87530-30-1) and has formula (la)

8. A medicament as claimed in any preceding claim, wherein metformin or a salt thereof and the compound of formula (I) or a salt thereof are combined in a single formulation.

9. The medicament according to claims 1 to 7, wherein metformin or a salt thereof and the compound of the general formula (I) or a salt thereof are formulated separately into a composition and used in combination.

10. A pharmaceutical composition comprising a medicament according to claims 1 to 9 in a pharmaceutically acceptable form.

11. The pharmaceutical composition according to claim 10, for use in the prevention or treatment of metabolic diseases or cardiovascular diseases.

12. The pharmaceutical composition for the use according to claim 11, wherein the metabolic disease is diabetes.

13. The pharmaceutical composition for the use according to claim 12, wherein the prevention or treatment of diabetes is by increasing inhibition of adipogenesis.

14. The pharmaceutical composition for the use according to claim 13, wherein the prevention or treatment of diabetes is improving insulin sensitivity by increasing inhibition of adipogenesis in liver cells and by activating AMP-activated protein kinase (AMPK).

15. The pharmaceutical composition for the use according to claims 9 to 14, wherein metformin is used in a sub-pharmacological dose to a subject.

16. The pharmaceutical composition for the use according to claim 15, wherein the sub-pharmacological dose does not exceed about 50% of the standard pharmacological dose for the subject.

17. The pharmaceutical composition for the use according to claims 14 and 15, wherein the sub-pharmacological dose of metformin reduces the incidence of side effects associated with metformin use, wherein the side effects are lactic acidosis and/or gastrointestinal side effects, in particular diarrhoea, nausea and vomiting.

18. A method of preventing or treating a metabolic disease or a cardiovascular disease, the method comprising administering to a subject in need thereof a medicament according to claims 1 to 9 or a pharmaceutical composition according to claims 10 to 17.

19. The method of claim 18, wherein the disease is diabetes.

Disclosure of Invention

Medicine

The present invention relates generally to a medicament comprising: (i) an agent suitable for the prevention or treatment of a metabolic disease or a cardiovascular disease, and (ii) a compound having the general formula (I) as described herein.

In one embodiment, compound (i) is an antidiabetic agent. In one embodiment, compound (i) is a biguanide compound, for example metformin or a salt thereof.

In particular, the present invention relates to a medicament comprising:

(i) metformin or a salt thereof; and

(ii) compounds having the general formula (I)

Or a salt thereof,

wherein, in compound (ii), R1, R2, R3, R4, R5, R6, R7, and R8 are each independently: h; OH; OMe; an O-glycoside; halogen; an aldehyde; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; a nitro group; a sulfonate ester; a sulfate ester; optionally substituted and/or optionally branched C1 to C20 alkyl; c2 to C20 alkenyl optionally substituted and/or optionally branched; a C4 to C20 polyalkenyl group optionally substituted and/or optionally branched; c2 to C20 alkynyl optionally substituted and/or optionally branched; or a C4 to C20 polyalkynyl group which is optionally substituted and/or optionally branched.

In some embodiments, in compound (ii), R1, R2, R3, R4, R7, and R8 are each independently: h; OH; OMe; an O-glycoside; halogen; an aldehyde; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; a nitro group; a sulfonate ester; a sulfate ester; optionally substituted and/or optionally branched C1 to C20 alkyl; c2 to C20 alkenyl optionally substituted and/or optionally branched; a C4 to C20 polyalkenyl group optionally substituted and/or optionally branched; c2 to C20 alkynyl optionally substituted and/or optionally branched; or C4 to C20 polyalkynyl optionally substituted and/or optionally branched; r5 is: h; OMe; an O-glycoside; halogen; an aldehyde; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; a nitro group; a sulfonate ester; a sulfate ester; optionally substituted and/or optionally branched C1 to C20 alkyl; c2 to C20 alkenyl optionally substituted and/or optionally branched; a C4 to C20 polyalkenyl group optionally substituted and/or optionally branched; c2 to C20 alkynyl optionally substituted and/or optionally branched; or C4 to C20 polyalkynyl optionally substituted and/or optionally branched; r6 is: h; OH; an O-glycoside; halogen; an aldehyde; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; a nitro group; a sulfonate ester; a sulfate ester; optionally substituted and/or optionally branched C1 to C20 alkyl; c2 to C20 alkenyl optionally substituted and/or optionally branched; a C4 to C20 polyalkenyl group optionally substituted and/or optionally branched; c2 to C20 alkynyl optionally substituted and/or optionally branched; or a C4 to C20 polyalkynyl group which is optionally substituted and/or optionally branched.

In some embodiments, in compound (ii), R1, R2, R3, R4, R7, and R8 are each independently: h; OH; OMe; an O-glycoside; halogen; an aldehyde; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; a nitro group; a sulfonate ester; a sulfate ester; r5 is: h; OMe; an O-glycoside; halogen; an aldehyde; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; a nitro group; a sulfonate ester; a sulfate ester; r6 is: h; OH; an O-glycoside; halogen; an aldehyde; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; a nitro group; a sulfonate ester; a sulfate ester.

In some embodiments, in compound (ii), R1, R2, R3, and R8 are each independently: h; OH; OMe; an O-glycoside; halogen; an aldehyde; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; a nitro group; a sulfonate ester; a sulfate ester; r4 is: h; OH; an O-glycoside; halogen; an aldehyde; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; a nitro group; a sulfonate ester; a sulfate ester; r5 is: h; OMe; an O-glycoside; halogen; an aldehyde; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; a nitro group; a sulfonate ester; a sulfate ester; r6 is: h; OH; an O-glycoside; halogen; an aldehyde; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; a nitro group; a sulfonate ester; a sulfate ester; and R7 is: h; OH; an O-glycoside; halogen; an aldehyde; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; a nitro group; a sulfonate ester; a sulfate ester.

In some embodiments, R1, R3, and R8 are each independently: h; OH; OMe; an O-glycoside; halogen; an aldehyde; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; a nitro group; a sulfonate ester; a sulfate ester; r2 is: OH; OMe; an O-glycoside; halogen; an aldehyde; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; a nitro group; a sulfonate ester; a sulfate ester; r4 is: h; OH; an O-glycoside; halogen; an aldehyde; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; a nitro group; a sulfonate ester; a sulfate ester; r5 is: h; OMe; an O-glycoside; halogen; an aldehyde; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; a nitro group; a sulfonate ester; a sulfate ester; r6 is: h; OH; an O-glycoside; halogen; an aldehyde; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; a nitro group; a sulfonate ester; a sulfate ester; and R7 is: h; OH; an O-glycoside; halogen; an aldehyde; a carboxylic acid; primary, secondary or tertiary amines; primary or secondary amides; a cyano group; a nitro group; a sulfonate ester; a sulfate ester.

In some embodiments, R1, R3, and R8 are each independently: h; OH; OMe; an O-glycoside; halogen; an aldehyde; a carboxylic acid; r2 is: OH; OMe; an O-glycoside; halogen; an aldehyde; a carboxylic acid; r4 is: h; OH; an O-glycoside; halogen; an aldehyde; a carboxylic acid; r5 is: h; OMe; an O-glycoside; halogen; an aldehyde; a carboxylic acid; r6 is: h; OH; an O-glycoside; halogen; an aldehyde; a carboxylic acid; and R7 is: h; OH; an O-glycoside; halogen; an aldehyde; a carboxylic acid.

In some embodiments, R1, R3, and R8 are each independently: h; OH; OMe; an O-glycoside; an aldehyde; a carboxylic acid; r2 is: OH; OMe; an O-glycoside; an aldehyde; a carboxylic acid; r4 is: h; OH; an O-glycoside; an aldehyde; a carboxylic acid; r5 is: h; OMe; an O-glycoside; an aldehyde; a carboxylic acid; r6 is: h; OH; an O-glycoside; an aldehyde; a carboxylic acid; and R7 is: h; OH; an O-glycoside; an aldehyde; a carboxylic acid.

In some embodiments, R1, R3, and R8 are each independently: h; OH; OMe; r2 is: OH; OMe; r4 is: h; OH; r5 is: h; OMe; r6 is: h; OH; and R7 is: h; and (5) OH.

In some embodiments, R1, R3, R6, and R8 are each independently: h; r2 is: OH; OMe; r4 is OH; r5 is: h; OMe; and R7 is: h; and (5) OH.

In one embodiment, compound (ii) is Lusianthridin (7-methoxy-9, 10-dihydrophenanthrene-2, 5-diol, CAS number 87530-30-1) and has formula

In one embodiment, compound (i) and compound (ii) of the medicament are combined in a single formulation. In one embodiment, compound (i) and compound (ii) of the medicament are combined in a single formulation with at least one other antidiabetic agent different from compound (i).

In one embodiment, compound (i) and compound (ii) of the medicament are formulated separately into a composition and used in combination. In one embodiment, compound (i) and compound (ii) of the medicament are formulated separately as a composition and used in combination with at least one other antidiabetic agent different from compound (i).

In some embodiments, the at least one other antidiabetic agent is a sulfonylurea compound or a salt thereof. In some embodiments, the at least one other antidiabetic agent is a thiazolidinedione compound or a salt thereof. In some embodiments, the at least one other antidiabetic agent is an alpha glucosidase inhibitor compound or a salt thereof. In some embodiments, the at least one other antidiabetic agent is a GLP-1 analog compound or salt thereof.

Pharmaceutical composition

Also provided is a pharmaceutical composition comprising a medicament of the invention as described herein, in a pharmaceutically acceptable form.

In some embodiments, the pharmaceutical composition comprises (I) an agent suitable for preventing or treating a metabolic disease or a cardiovascular disease, or a pharmaceutically acceptable salt thereof, and (ii) a compound having the general formula (I) or a pharmaceutically acceptable salt thereof as described herein.

In some embodiments, compound (i) is an antidiabetic agent. In some embodiments, the antidiabetic agent is a biguanide or a pharmaceutically acceptable salt thereof. In some embodiments, the biguanide is metformin or a pharmaceutically acceptable salt thereof.

In some embodiments, in compound (ii), R1, R3, and R8 are each independently: h; OH; OMe; an O-glycoside; halogen; an aldehyde; a carboxylic acid; r2 is: OH; OMe; an O-glycoside; halogen; an aldehyde; a carboxylic acid; r4 is: h; OH; an O-glycoside; halogen; an aldehyde; a carboxylic acid; r5 is: h; OMe; an O-glycoside; halogen; an aldehyde; a carboxylic acid; r6 is: h; OH; an O-glycoside; halogen; an aldehyde; a carboxylic acid; and R7 is: h; OH; an O-glycoside; halogen; an aldehyde; a carboxylic acid.

In some embodiments, R1, R3, and R8 are each independently: h; OH; OMe; an O-glycoside; an aldehyde; a carboxylic acid; r2 is: OH; OMe; an O-glycoside; an aldehyde; a carboxylic acid; r4 is: h; OH; an O-glycoside; an aldehyde; a carboxylic acid; r5 is: h; OMe; an O-glycoside; an aldehyde; a carboxylic acid; r6 is: h; OH; an O-glycoside; an aldehyde; a carboxylic acid; and R7 is: h; OH; an O-glycoside; an aldehyde; a carboxylic acid.

In some embodiments, R1, R3, and R8 are each independently: h; OH; OMe; r2 is: OH; OMe; r4 is: h; OH; r5 is: h; OMe; r6 is: h; OH; and R7 is: h; and (5) OH.

In some embodiments, R1, R3, R6, and R8 are each independently: h; r2 is: OH; OMe; r4 is OH; r5 is: h; OMe; and R7 is: h; and (5) OH.

In some embodiments, the pharmaceutical composition comprises (i) metformin or a pharmaceutically acceptable salt thereof, and (ii) Lusianthridin or a pharmaceutically acceptable salt thereof.

In one embodiment, compound (i) and compound (ii) of the pharmaceutical composition are combined in a single formulation. In one embodiment, compound (i) and compound (ii) of the pharmaceutical composition are combined in a single formulation with at least one other antidiabetic agent different from compound (i).

In one embodiment, compound (i) and compound (ii) of the pharmaceutical composition are formulated separately into a composition and used in combination. In one embodiment, compound (i) and compound (ii) of the pharmaceutical composition are formulated separately as a composition and used in combination with at least one other antidiabetic agent different from compound (i).

In some embodiments, the at least one other antidiabetic compound is a sulfonylurea compound or a pharmaceutically acceptable salt thereof. In some embodiments, the at least one other antidiabetic agent is a thiazolidinedione compound or a pharmaceutically acceptable salt thereof. In some embodiments, the at least one other antidiabetic agent is an alpha glucosidase inhibitor compound or a pharmaceutically acceptable salt thereof. In some embodiments, the at least one other anti-diabetic agent is a GLP-1 analog compound or a pharmaceutically acceptable salt thereof.

The invention also provides a pharmaceutical composition as described herein for use in the prevention or treatment of a metabolic disease or a cardiovascular disease.

In one embodiment, the metabolic disease is diabetes. In one embodiment, the metabolic disease is non-alcoholic fatty liver disease (NAFLD). In one embodiment, the metabolic disease is non-alcoholic steatohepatitis (NASH).

In one embodiment, the pharmaceutical composition is for preventing or treating diabetes or NAFLD by increasing inhibition of adipogenesis.

In one embodiment, the pharmaceutical composition is for preventing or treating diabetes or NAFLD by increasing inhibition of adipogenesis in hepatocytes.

In one embodiment, the pharmaceutical composition is for preventing or treating diabetes by (i) increasing inhibition of adipogenesis in hepatocytes and (ii) improving insulin sensitivity by activating AMP-activated protein kinase (AMPK).

In one embodiment, compound (i) when used in combination with compound (ii) may be used at a lower than pharmacological dose compared to a pharmaceutical composition comprising a pharmacological dose of compound (i) but in which compound (ii) is absent, for substantially the same degree of adipogenesis inhibition.

In one embodiment, the pharmaceutical composition is for use according to the invention, wherein the subject is a human.

In some embodiments, the pharmaceutical composition is for use according to the invention, wherein the subject has or is at risk of developing a further condition, disorder or disease, wherein the further condition, disorder or disease relates to cardiometabolic health, cancer, obesity, non-alcoholic fatty liver disease and/or cardiovascular disease.

In some embodiments, the pharmaceutical composition is for use according to the invention, wherein the further condition, disorder or disease is related to cardiometabolic health and/or cancer.

In one embodiment, the pharmaceutical composition is for use according to the invention, wherein compound (i) is used in a sub-pharmacological dose.

In one embodiment, the pharmaceutical composition is for use according to the invention, wherein metformin is used in a sub-pharmacological dose which is not less than about 50% of the standard pharmacological dose for a subject.

In some embodiments, the pharmaceutical composition is used according to the invention, wherein the sub-pharmacological dose of metformin reduces the incidence of side effects associated with metformin use, and wherein the side effects are lactic acidosis and/or gastrointestinal side effects, in particular diarrhea, nausea and vomiting.

In some embodiments, the pharmaceutical composition for use according to the invention is a Glucophage or Glucophage XR. A typical standard pharmacological dose for Glucophage is 1g twice daily. Typical standard pharmacological doses for Glucophage XR are up to 2g, taken once daily with dinner.

In one embodiment, compound (i) and compound (ii) are combined in a single formulation. In one embodiment, compound (i) and compound (ii) are combined in a single formulation with at least one other antidiabetic agent different from compound (i).

In one embodiment, compound (i) and compound (ii) are formulated separately into a composition and used in combination. In one embodiment, compound (i) and compound (ii) are formulated separately as a composition and used in combination with at least one other antidiabetic agent different from compound (i).

In some embodiments, the at least one additional antidiabetic agent is a sulfonylurea compound or a pharmaceutically acceptable salt thereof. In some embodiments, the at least one other antidiabetic agent is a thiazolidinedione compound or a pharmaceutically acceptable salt thereof. In some embodiments, the at least one other antidiabetic agent is an alpha glucosidase inhibitor compound or a pharmaceutically acceptable salt thereof. In some embodiments, the at least one other anti-diabetic agent is a GLP-1 analog compound or a pharmaceutically acceptable salt thereof.

Methods of preventing or treating disease

The present invention also provides a method of preventing or treating a metabolic disease or a cardiovascular disease, the method comprising administering to a subject in need thereof a medicament or pharmaceutical composition of the invention, wherein the medicament or pharmaceutical composition comprises (I) an agent suitable for preventing or treating a metabolic disease or a cardiovascular disease and/or (ii) a compound having the general formula (I) as described herein.

In one embodiment, the metabolic disease is diabetes. In one embodiment, the method increases inhibition of adipogenesis, for example, in hepatocytes. In one embodiment, the method improves insulin sensitivity by activating AMP-activated protein kinase (AMPK).

In one embodiment, compound (i), when used in combination with compound (ii), may be administered at a lower than pharmacological dose compared to a pharmaceutical composition comprising a pharmacological dose of compound (i) but in which compound (ii) is absent, for substantially the same degree of adipogenesis inhibition.

In some embodiments, the subject has or is at risk of developing an additional condition, disorder or disease, wherein the additional condition, disorder or disease involves cardiometabolic health, cancer, obesity, non-alcoholic fatty liver disease and/or cardiovascular disease. In some embodiments, the additional condition, disorder or disease is related to cardiometabolic health and/or cancer.

In one embodiment, compound (i) is administered at a sub-pharmacological dose.

In one embodiment, compound (i) is metformin, wherein metformin is administered in a sub-pharmacological dose which is no more than about 50% of the standard pharmacological dose for a subject.

In some embodiments, the sub-pharmacological dose of metformin reduces the incidence of side effects associated with metformin administration, and wherein the side effects are lactic acidosis and/or gastrointestinal side effects, particularly diarrhea, nausea and vomiting.

The antidiabetic agent metformin as disclosed herein may be in the form of its hydrochloride saltIs sold by Bristol-Myers Squibb Company. In some embodiments, the pharmaceutical composition for use according to the invention is a Glucophage or Glucophage XR. A typical standard pharmacological dose for Glucophage is 1g twice daily. Typical standard pharmacological doses for Glucophage XR are up to 2g, taken once daily with dinner.

In one embodiment, compound (i) and compound (ii) are combined in a single formulation. In one embodiment, compound (i) and compound (ii) are combined in a single formulation with at least one other antidiabetic agent different from compound (i).

In one embodiment, compound (i) and compound (ii) are formulated separately into compositions and administered in combination. In one embodiment, compound (i) and compound (ii) are formulated separately as a composition and administered in combination with at least one other antidiabetic agent different from compound (i).

In some embodiments, the at least one other antidiabetic compound is a sulfonylurea compound or a pharmaceutically acceptable salt thereof. In some embodiments, the at least one other antidiabetic agent is a thiazolidinedione compound or a pharmaceutically acceptable salt thereof. In some embodiments, the at least one other antidiabetic agent is an alpha glucosidase inhibitor compound or a pharmaceutically acceptable salt thereof. In some embodiments, the at least one other anti-diabetic agent is a GLP-1 analog compound or a pharmaceutically acceptable salt thereof.

The invention also provides a method for first line treatment of a metabolic disease in a subject, the method comprising administering to a subject in need of treatment a drug as first line treatment, the drug being a therapeutically effective dose of a pharmaceutical formulation comprising a sub-pharmacological dose of metformin and a compound of formula (I) as described herein, the compound being an AMPK activator.

In one embodiment, the metabolic disease is diabetes. In one embodiment, the human subject is non-medicated.

Drawings

FIG. 1: AMP and lucianthridin activation of bacterially expressed AMPK α 2 β 1 γ 1. The combination of Lusianthridin (7-methoxy-9, 10-dihydrophenanthrene-2, 5-diol, CAS number 87530-30-1) with AMP leads to additive activation of AMPK. AMP binding by the AMPK γ subunit causes a dose-dependent increase in AMPK activity, and the addition of various concentrations of lucianthridin causes cumulative activation of AMPK without affecting EC50 of AMP activation.

FIG. 2: lusianthridin alone and metformin alone at low concentrations caused inhibition of adipogenesis. When these two activators are used in combination, lipogenesis is further increased.

Examples

Example 1

Lusianthridin causes additive effect on AMP activation of AMPK alpha 2 beta 1 gamma 1 complex which activates bacterial expression。

AMPK heterotrimers are expressed in bacteria and purified by His-alpha subunit using nickel purification, further purified by gel filtration, and finally phosphorylated by incubation with CaMKK β. The phosphorylated AMPK was incubated with different concentrations of AMP for 30 minutes using substrates and reagents from the HTRF-KinEASE Cisbio assay kit (STK S1 kit). Phosphorylation of the substrate was measured by incubation with donor and acceptor antibodies for 2 hours at room temperature according to the manufacturer's protocol (and Coulerie et al, 2016, see below), and phosphorylated peptides were detected by performing HTRF. The 665nm/620nm ratio was determined and shown in the graph. In the presence of a fixed concentration of Lusianthridin, AMP has an additive activation of AMPK.

Reference 1: standardized LC x LC-ELSD fractionation procedure for the identification of trace bioactive agents via enzymatic screening of natural extracts. Coulerie P, Ratinaud Y, Moco S, Melminod L, Naranjo Pinta M, Boccard J, Buttot L, Deak M, Sakamoto K, Queiroz EF, Wolfinder JL, Barron D., "journal of Natural products" (J Nat Prod.), 11/23/2016, vol 79, 11/64, 2856, 2864. Electronically released 10 months and 28 days in 2016.

Example 2

Lusianthridin and metformin have additive effects on inhibition of adipogenesis in primary hepatocytes。

And (3) separating the liver cells: the liver was first perfused with 50ml of perfusion buffer (Krebs-Hepes buffer containing 0.5. mu.M EDTA), followed by 50ml of collagenase A buffer (containing 5mM CaCl)₂And 0.5mg/ml collagenase in Krebs-Hepes buffer). After passing through a 100 μ M mesh, the cell solution was washed several times with cold medium and finally the cell culture pellet was resuspended in medium (medium 199(M199) + GlutaMAX, 100U/ml penicillin G and 100 μ G/ml streptomycin, 0.1% (wt/vol) BSA, 10% FCS, 10nM insulin, 200nM triiodothyronine and 500nM dexamethasone). Hepatocyte attachment was retained (3-4 hours) and cultured overnight in M199 supplemented with antibiotics and 100nM dexamethasone. The next morning cells were used for the experiment.

For adipogenesis measurements in primary hepatocytes, cells were seeded in 6-well plates at 600K cells/well overnight. In the presence of [1-¹⁴C]In the case of acetate, the medium was replaced with fresh M199 medium alone for 2 hours before incubation at 37 ℃ for 1 hour with different concentrations of Lusianthridin or Compound 1. After separation from the aqueous phase, the [ 2 ] is determined in the lower organic layer¹⁴C]Incorporation into fatty acids. The results are shown as Disintegration Per Minute (DPM) per μ g protein.

Adipogenesis is controlled by the AMPK substrate ACC, and phosphorylation and inhibition of ACC by AMPK leads to a reduction in adipogenesis. By determining¹⁴Incorporation of C-labeled acetate into fatty acids, adipogenesis in primary hepatocytes was measured. Adipogenesis was monitored for 1 hour at 37 ℃ in the presence or absence of low/non-saturating concentrations of Lusianthridin or metformin. Furthermore, cells were incubated in the presence of both compounds. The results shown in FIG. 2 show that Lusianthridin and metformin were able to inhibit adipogenesis% (as compared to control untreated cells)Ratio). Lusianthridin and metformin, when combined, cause further inhibition of adipogenesis. These data indicate that when combined with low concentrations of AMPK activators, there is greater inhibition of adipogenesis than the action of these activators on their own.

Detailed Description

The term "alkyl" refers to a branched or straight saturated hydrocarbon chain having 1 to 20 carbon atoms, or 1 to 15 carbon atoms, or 1 to 10 carbon atoms, or 1 to 8 carbon atoms, or 1 to 6 carbon atoms, or 1 to 4 carbon atoms. The term is exemplified by groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, n-hexyl, n-decyl, tetradecyl.

The term "substituted alkyl" refers to:

1) an alkyl chain as defined above having 1, 2, 3, 4 or 5 substituents (in some embodiments, 1, 2 or 3 substituents) selected from alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkenyl, cycloalkoxy, cycloalkenyloxy, acyl, amido, acyloxy, amino, substituted amino, aminocarbonyl, alkoxycarbonylamino, azido, cyano, halogen, hydroxy, keto, thiocarbonyl, carboxy, carboxyalkyl, arylthio, heteroarylthio, heterocyclylthio, thiol, alkylthio, aryl, aryloxy, heteroaryl, aminosulfonyl, aminocarbonylamino, heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino, nitro, -S (O) -alkyl, -S (O) -cycloalkyl, -S (O) -heterocyclyl, amino, nitro, amino, cycloalkyl, amino, -S (O) -aryl, -S (O) -heteroaryl, -S (O) 2-alkyl, -S (O) 2-cycloalkyl, -S (O) 2-heterocyclyl, -S (O) 2-aryl and-S (O) 2-heteroaryl. Unless otherwise constrained by the definition, all substituents may optionally be further substituted by 1, 2 or 3 substituents selected from alkyl, alkenyl, alkynyl, carboxyl, carboxyalkyl, aminocarbonyl, hydroxyl, alkoxy, halogen, CF3, amino, substituted amino, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl and-s (o) n R < a >, wherein R < a > is alkyl, aryl or heteroaryl and n is 0, 1 or 2; or

2) Interrupted by an alkyl chain of 1 to 10 atoms (e.g. 1, 2, 3, 4 or 5 atoms) as defined above, these atoms being independently selected from oxygen, sulphur and NR < a >, wherein R < a > is selected from hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, heteroaryl and heterocyclyl. All substituents may be optionally further substituted with alkyl, alkenyl, alkynyl, carboxy, carboxyalkyl, aminocarbonyl, hydroxy, alkoxy, halogen, CF3, amino, substituted amino, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl and-s (o) n R < a > wherein R < a > is alkyl, aryl or heteroaryl and n is 0, 1 or 2; or

3) An alkyl chain as defined above having 1, 2, 3, 4 or 5 substituents as defined above and further interrupted by 1 to 10 atoms (e.g. 1, 2, 3, 4 or 5 atoms) as defined above.

The term "alkenyl" refers to a class of alkyl chains in which two atoms of the alkyl group form a double bond that is not part of an aromatic group. That is, the alkenyl chain comprises the pattern R-c (R) ═ c (R) -R, where R refers to the remainder of the alkenyl group, which may be the same or different. Non-limiting examples of alkenyl chains include-CH ═ CH2, -C (CH3) ═ CH2, -CH ═ CHCH3, -C (CH3) ═ CHCH3, -CH2-CH ═ C (CH3)2, and-C (CH3)2-CH ═ CH 2. The alkenyl moiety may be branched, straight-chain or cyclic (in which case it will also be referred to as a "cycloalkenyl" group). The alkenyl group may be optionally substituted.

The term "alkynyl" refers to a class of alkyl chains in which two atoms of the alkyl group form a triple bond. That is, alkynyl groups contain the pattern R-C ≡ C-R, where R refers to the remainder of the alkynyl group, which may be the same or different. Non-limiting examples of alkynyl groups include-C ≡ CH, -C ≡ CCH3 and-C ≡ CCH2CH 3. The "R" moiety of the alkynyl moiety may be branched, straight chain or cyclic. Alkynyl groups may be optionally substituted.

The term "polyunsaturated" means

1) Wherein more than one pair of atoms of an alkyl group forms a chain known as a polyalkenyl group that is not a double bond part of an aromatic group. That is, the alkenyl chain comprises a number of R-c (R) ═ c (R) -R patterns, where R refers to the remainder of the alkenyl group, which may be the same or different. Non-limiting examples of polyalkenyl chains include-CH-, -CH 2-CH-CCH 3-CH2-CH 2-CH-C (CH3)2 and-CH 2-CH-CCH 3-CH2-CH 2-CH-CCH 3-CH2-CH 2-CH-C (CH3) 2. The polyalkenyl moiety may be branched or straight chain. The polyalkenyl moiety comprising two double bonds may be cyclic (in which case it will also be referred to as a "cycloalkadienyl" group). Non-limiting examples of cycloalkadienyl groups include cyclopentadiene and cyclohexadiene groups. The polyalkenyl group may be optionally substituted.

2) Chains known as poly-alkynyl in which more than one pair of atoms of the alkyl group forms a triple bond. That is, the polyacetynyl group comprises several patterns R-C.ident.C-R, wherein R refers to the remainder of the alkynyl group, which may be the same or different. Non-limiting examples of polyalkynyl groups include-CH 2-CH 2-C-CH. The "R" moiety of the polyalkynyl moiety can be branched, straight chain, or cyclic. Alkynyl groups may be optionally substituted.

3) One type of alkyl chain in which at least one pair of atoms of the alkyl group forms a double bond and one pair of atoms of the alkyl group forms a triple bond. That is, the polyunsaturated chain comprises a pattern of R-C (R) ═ C (R) -R and R-C ≡ C-R, where R refers to the remainder of the polyunsaturated chain, which may be the same or different. Non-limiting examples of this type of polyunsaturated chain include-CH 2-CH ═ CH-C ≡ CH. The "R" moiety of the polyunsaturated moiety can be branched, linear, or cyclic. The polyunsaturated chain may be optionally substituted.

4) The interjacent as defined in paragraphs 1-3 above is interrupted by a polyunsaturated chain of 1 to 10 atoms (e.g., 1, 2, 3, 4 or 5 atoms) independently selected from the group consisting of oxygen, sulfur and NR < a >, wherein R < a > is selected from the group consisting of hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, heteroaryl and heterocyclyl.

As used herein, the term "ring" refers to any covalently closed structure. Rings include, for example, carbocycles (e.g., aryl and cycloalkyl), heterocycles (e.g., heteroaryl and non-aromatic heterocycle), aromatics (e.g., aryl and heteroaryl), and non-aromatics (e.g., cycloalkyl and non-aromatic heterocycle). The rings may be optionally substituted. The ring may form part of a ring system. As used herein, the term "ring system" refers to two or more rings, wherein two or more rings are fused. The term "fused" refers to a structure in which two or more rings share one or more bonds.

The term "halogen atom" may refer to a fluorine atom, a chlorine atom, a bromine atom or an iodine atom.

As used herein, the term "analog" is understood to mean a compound having a structure similar to another structure, but differing therefrom in certain components. A "derivative" is a compound that is produced by substituting one or more atoms with another atom or group of atoms or that is actually synthesized from the parent compound.

The components of the chemical structures described herein may be further defined as follows: the term "unsaturated" means that it contains at least one, up to eight, double bonds between carbon atoms. "dehydration" refers to the loss of water between two adjacent carbons, one carbon bearing a hydroxyl group and the other carbon bearing at least one hydrogen, resulting in the formation of a double bond. "reducing" refers to the addition of hydrogen to a double bond, resulting in the formation of a single bond, typically the reduction of a carbonyl to an alcohol, or the reduction of an unsaturated chain to a saturated chain. Carbon oxidation can be, for example, stepwise from methyl to alcohol, to aldehyde, and finally to carboxylic acid.

As used herein, the term "aldehyde" denotes an organic compound (n is 1 or greater) having the general structure-C- [ C (═ O) ] n-H or H- [ C (═ O) ] n-H, and wherein the carbon atom bonded to the- [ C (═ O) ] -n group is not double bonded to oxygen, sulfur, selenium or tellurium, or triple bonded to nitrogen.

As used herein, the term "amine" refers to an organic compound having a nitrogen atom that is mono-or double-bonded to a carbon atom and wherein the carbon atom bonded to the nitrogen atom lacks double-bonding to oxygen, sulfur, selenium, or tellurium or triple-bonding to nitrogen. Furthermore, those compounds in which the same nitrogen atom is bonded to a-C (═ X) -group (X is O, S, Se or Te) and to a carbon atom which is not doubly bonded to oxygen, sulfur, selenium or tellurium are not to be regarded as amines, for example-C-NH-C (X ═) -

As used herein, the term "cyano" refers to a triple bond between adjacent carbon and nitrogen atoms.

As used herein, the term "carboxylic acid" denotes the presence of a-C (═ O) OH group.

As used herein, the term "metformin" refers to metformin or its salts such as the hydrochloride, metformin (2:1) fumarate and metformin (2:1) succinate, hydrobromide, p-chlorophenoxyacetate or pamoate, and other known metformin salts of mono-and dicarboxylic acids, all of which are collectively referred to as metformin. Metformin as referred to herein may be a metformin hydrochloride salt, i.e., asMetformin hydrochloride salt is sold (trade mark of Bristol-Myers Squibb Company).

The pharmaceutical composition of the present invention may be formulated into oral unit dosage forms, such as tablets, pills, pellets, capsules, powders, lozenges, granules, solutions, suspensions, emulsions, syrups, elixirs, oral liquid preparations, edible food products, prebiotics, probiotics, beverages, food bars, milkshakes, smoothies, or yogurts. It may also contain one or more optional agents selected from the group consisting of sweetening agents, flavouring agents, colouring agents, preserving agents, time-delay or delayed disintegration materials, standard oral carriers, suitable carriers, excipients and diluents.

It may optionally contain one or more fillers or excipients such as lactose, sugar, corn starch, modified corn starch, mannitol, sorbitol, inorganic salts such as calcium carbonate and/or cellulose derivatives such as lignocellulose and microcrystalline cellulose.

One or more binders may be present in addition to or in place of the filler. Examples of such binders suitable for use herein include polyvinylpyrrolidone, lactose, starches such as corn starch, modified corn starch, sugar, gum arabic, and the like, and wax binders in the form of fine powders such as carnauba wax, paraffin wax, spermaceti wax, polyethylene, or microcrystalline wax.

Where the pharmaceutical composition is in the form of a tablet, it will contain one or more tableting lubricants such as magnesium stearate, stearic acid, palmitic acid, calcium stearate, talc, carnauba wax and the like. Other conventional ingredients that may optionally be present include preservatives, stabilizers, anti-adherents or silica flow conditioners or glidants, such as Syloid brand silica and FD & C pigments.

The tablet may also include a coating layer. The coating layer (applied on the outer solid phase comprising the inner solid phase particles embedded therein) may comprise any conventional coating formulation and will comprise: one or more film formers or binders such as hydrophilic polymers, e.g., hydroxypropyl methylcellulose, and/or hydrophobic polymers, e.g., methacrylate neutral polymers, ethylcellulose, cellulose acetate, polyvinyl alcohol-maleic anhydride copolymers, β -pinene polymers, glycerol esters of wood resins, and the like; and one or more plasticizers such as triethyl citrate, diethyl phthalate, propylene glycol, glycerin, butyl phthalate, castor oil, and the like. Both the core and the coating formulation may comprise aluminium lakes to provide colour.

The film-forming agent is applied from a solvent system comprising one or more solvents, the one or more solvents comprising: water; alcohols such as methanol, ethanol or isopropanol; ketones such as acetone or ethyl methyl ketone; chlorinated hydrocarbons, such as dichloromethane, dichloroethane and 1,1, 1-trichloroethane.

In the case of pigments, the pigment will be applied with a film former, plasticizer and solvent composition.

The finished dosage form may be a compressed tablet or a hard gelatin capsule, preferably a tablet. The tablets may optionally be film coated. The total amount of drug per dosage unit will be such as to provide the patient with a conveniently sized dosage form.

The pharmaceutical composition of the present invention is preferably administered orally. In some embodiments, the pharmaceutical composition may be administered by: intravenous administration, topical administration, parenteral administration, intraperitoneal administration, intramuscular injection administration, intrathecal injection administration, intralesional administration, intracranial administration, intranasal administration, intraocular administration, intracardiac administration, intravitreal administration, intraosseous administration, intracerebral administration, intraarterial administration, intraarticular administration, intradermal administration, transdermal administration, transmucosal administration, sublingual administration, enteral administration, sublabial administration, insufflation administration, suppository administration, inhalation administration, or subcutaneous administration.

The compositions of the present invention may have an acute effect that may be observed in less than one month. Additionally or alternatively, the composition may have a long-term effect, and thus various embodiments include administering the composition to an individual (e.g., orally) for a period of at least one month; preferably at least two months, more preferably at least three months, four months, five months or six months; most preferably at least one year. During this period of time, the composition may be administered to the individual at least one day per week; preferably at least two days per week, more preferably at least three, four, five or six days per week; most preferably seven days per week. The composition may be administered in a single dose per day or in multiple individual doses per day.

As used herein, "AMPK activator" refers to a compound that increases phosphorylation of a downstream substrate of AMPK (phosphorylated or non-phosphorylated) and/or a compound that increases phosphorylation or enzymatic activity of AMPK.

As used herein, a "direct AMPK activator" refers to a compound that activates AMPK via direct interaction (through at least one of its subunits binding to the subunit) with the AMPK trimeric complex structure.

As used herein, a condition, disorder or disease that "responds to AMPK activation" refers to a condition, disorder or disease in which symptoms will be alleviated by activation of AMPK or the course of which will be favorably altered, including but not limited to cancer, NAFLD, metabolic disorders, diabetes, dyslipidemia, hypertension, overweight and obesity.

As used herein, the term "diabetes" includes insulin-dependent diabetes (i.e., IDDM, also known as type 1 diabetes), non-insulin-dependent diabetes (i.e., NIDDM, also known as type 2 diabetes), and prediabetes. Type 1 diabetes is the result of an absolute deficiency in insulin, a hormone that regulates glucose utilization. Type 2 diabetes usually occurs in the face of normal or even elevated levels of insulin and appears to be the result of the inability of tissues to respond properly to insulin. This is called "insulin resistance". Most type 2 diabetic patients are also overweight or obese. One of the criteria for diagnosing diabetes is fasting plasma glucose levels. The diabetic subject has a fasting plasma glucose level greater than or equal to 126 mg/dl. A pre-diabetic subject is a person with pre-diabetes. The pre-diabetic subject is a subject with impaired fasting glucose (fasting plasma glucose levels greater than or equal to 100mg/dl and less than 126 mg/dl); or impaired glucose tolerance (2 hours plasma glucose levels ≥ 140mg/dl and less than 200 mg/dl); or subjects with insulin resistance that result in an increased risk of developing diabetes. Prevention of type 2 diabetes includes treatment of prediabetes.

As used herein, the term "dyslipidemia" encompasses abnormal levels of any lipid fraction as well as specific lipoprotein abnormalities. For example, it refers to an increase in plasma cholesterol and/or an increase in triglycerides and/or an increase in free fatty acids and/or a low High Density Lipoprotein (HDL) level and/or a high Low Density Lipoprotein (LDL) level and/or a high Very Low Density Lipoprotein (VLDL) level. Dyslipidemia can, for example, lead to the development of atherosclerosis and ultimately symptomatic vascular disease, including coronary heart disease. Dyslipidemia may or may not be associated with diabetes.

As used herein, the term "metabolic disorder" encompasses any abnormal chemical and enzymatic reactions that disrupt normal metabolism due to environmental and genetic factors (environmental factors including physical activity, nutrition), resulting in excessive levels or deficiency of certain substances and dysfunction of energy homeostasis. Non-limiting examples of metabolic disorders include diabetes, NAFLD, dyslipidemia, hypertension, overweight, obesity, and any combination thereof.

As used herein, "AMPK-related diseases" include pathological or pathogenic conditions in which activation of AMPK provides a beneficial effect. Examples of such diseases or disorders include obesity, diabetes, metabolic syndrome, acute inflammatory lung injury, heart disease, reperfusion ischemia, cancer, aging, retinal degeneration, cardiac hypertrophy, non alcoholic fatty liver disease, hypertension, albuminuria, sporadic alzheimer's disease, muscular dystrophy, and osteoarthritis.

"preventing" or "prevention" includes reducing the risk and/or severity of a condition, disorder or disease.

The terms "treat," "treating," "alleviate," and "alleviate" include both prophylactic or preventative treatment (prevention and/or slowing of the development of the pathological condition or disorder of interest) and curative, therapeutic, or disease modifying treatment, including curative, slowing, lessening the symptoms of, and/or interrupting the progression of, a diagnosed pathological condition or disorder; and includes treating patients at risk of contracting a disease or suspected of contracting a disease, as well as treating patients who are ill or have been diagnosed as having a disease or medical condition. The term does not necessarily mean that the subject is treated until complete recovery. These terms also refer to health maintenance and/or promotion in a subject who does not have a disease but may be susceptible to developing an unhealthy condition. These terms are also intended to include the enhancement or otherwise enhancement of one or more primary preventative or therapeutic measures. The terms "treat," "alleviate," and "alleviate" are also intended to include dietary management of a disease or condition or dietary management of preventing or preventing a disease or condition. The treatment may be patient-related or physician-related.

Obesity is an excess of body fat relative to lean body mass, a chronic disease that is highly prevalent in modern society. It is associated not only with social perseveration (social stigma), but also with shortened lifespan and a number of Medical problems, including adverse mental development, coronary artery disease, hypertension, stroke, diabetes, hyperlipidemia and some cancers (see, e.g., Nishina, et al., Metab.43: 554-.

"obesity-related disorder" refers to a disease or condition that is overweight or has an excessive "Body Mass Index (BMI)" associated with the progression or inhibition of the disease or condition. Representative examples of obesity-related disorders include, but are not limited to, diabetes, diabetic complications, insulin sensitivity, polycystic ovary disease, hyperglycemia, dyslipidemia, insulin resistance, metabolic syndrome, obesity, weight gain, inflammatory diseases, digestive organ diseases, angina pectoris, myocardial infarction, angina pectoris or sequelae of myocardial infarction, senile dementia and cerebrovascular dementia. See Harrison's Principles of Internal Medicine,13th Ed., McGraw Hill Companies Inc., New York (1994). Examples of inflammatory disorders include, but are not limited to, diseases of the digestive organ (such as ulcerative colitis, crohn's disease, pancreatitis, gastritis, benign tumors of the digestive organ, digestive polyps, hereditary polyp syndrome, colon cancer, rectal cancer, stomach cancer, and ulcerative diseases of the digestive organ), angina pectoris, myocardial infarction, angina pectoris or sequelae of myocardial infarction, senile dementia, cerebrovascular dementia, immune diseases, and cancer in general.

The term "subject" or "individual" means any animal, including humans, that may benefit from one or more of the compounds, compositions, or methods disclosed herein. Generally, the subject is a human or avian animal, bovine, canine, equine, feline, caprine, wolf, murine, ovine, and porcine animal. The subject may be a "companion animal," which is any domesticated animal and includes, but is not limited to, cats, dogs, rabbits, guinea pigs, ferrets, hamsters, mice, gerbils, horses, cows, goats, sheep, donkeys, pigs, and the like. Preferably, the subject is a human or a companion animal, such as a dog or cat. The subject may be elderly or elderly. In the human context, the term "elderly" means at least 60 years of age, preferably 63 years of age or older, more preferably 65 years of age or older, and most preferably 70 years of age or older, from birth. In the context of humans, the term "elderly" refers to an age of at least 45 years, preferably over 50 years, more preferably over 55 years, from birth and includes elderly individuals. For other animals, "elderly" means having exceeded 50% of the average lifespan of their particular species and/or species within. Animals are considered "elderly" if they exceed 66% of the average life expectancy, preferably over 75% of the average life expectancy, more preferably over 80% of the average life expectancy. The senior cat or dog is at least about 7 years of age from birth.

As used herein, an "effective dose" or "effective amount" is an amount that prevents a deficiency, treats a disorder, condition, or disease in a subject, or more generally, reduces symptoms, manages disease progression, or provides a nutritional, physiological, or medical benefit to a subject. The relative terms "improve," "increase," "enhance," and the like refer to the effect of a composition disclosed herein relative to a composition lacking one or more ingredients and/or having a different amount of one or more ingredients, but otherwise being the same.

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 component" or "the component" includes two or more components.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Reference is made herein to various methods and materials known to those skilled in the art. Standard references describing the general principles of recombinant DNA technology include Sambrook et al, Molecular Cloning, A Laboratory Manual, 2 nd edition, Cold Spring Harbor Laboratory Press, New York (1989); kaufman et al, eds., Handbook of Molecular and Cellular Methods in Biology in Medicine, CRC Press, Boca Raton (1995); McPherson, Ed., Directed Mutagenesis, A Practical Approach, IRL Press, Oxford (1991). Standard references which illustrate The general principles of pharmacology include The Pharmacological Basis of Therapeutics, 10 th edition, McGraw Hill Companies Inc., New York (2001), Goodman and Gilman. Standard Medical terms used herein have the meaning defined in Stedman's Medical Dictionary, 27 th edition, with veterinary inserts.

All percentages expressed herein are by weight of the total weight of the composition, unless otherwise indicated. As used herein, "about" and "substantially" are understood to mean a number within a range of values, for example in the range of-10% to + 10% of the number referred to, preferably-5% to + 5% of the number referred to, more preferably-1% to + 1% of the number referred to, most preferably-0.1% to + 0.1% of the number referred to. All numerical ranges herein should be understood to include all integers or fractions within the range. Additionally, these numerical ranges should be understood to provide support for claims directed to any number or subset of numbers within the range. For example, a disclosure of 1 to 10 should be understood to support a range of 1 to 8, 3 to 7,1 to 9, 3.6 to 4.6, 3.5 to 9.9, and so forth.

As used in this specification, the terms "comprises" and "comprising," whether in the transitional phrase or in the context of the claims, are to be construed in an open-ended manner. That is, the term is to be interpreted synonymously with the phrases "having at least" or "including at least". The term "comprising," when used in the context of a process, means that the process includes at least the recited steps, but may include additional steps. The term "comprising" when used in the context of a compound or composition means that the compound or composition includes at least the recited feature or compound, but may also include additional features or compounds. The term "and/or" as used in the context of "X and/or Y" should be interpreted as "X" or "Y" or "X and Y". As used herein, the terms "example" and "such as" (especially when followed by a list of terms) are merely exemplary and illustrative and should not be considered exclusive or comprehensive.

Reference will now be made in detail to specific embodiments of the invention. While the invention will be described in conjunction with these specific embodiments, it will be understood that it is not intended to limit the invention to such specific embodiments. On the contrary, it is intended to cover alternatives, modifications, and equivalents as may be included within the spirit and scope of the invention as defined by the appended claims. In the description, numerous specific details are set forth in order to provide a thorough understanding of the present invention. The present invention may be practiced without some or all of these specific details. In other instances, well known methods and protocols have not been described in detail so as not to unnecessarily obscure the present invention.