CN111655275A

CN111655275A - Compositions, kits and methods for treating type II diabetes

Info

Publication number: CN111655275A
Application number: CN201880080992.XA
Authority: CN
Inventors: 王沛然
Original assignee: Reno Huasheng Medical Co ltd
Current assignee: Reno Huasheng Medical Co ltd
Priority date: 2017-12-18
Filing date: 2018-12-17
Publication date: 2020-09-11
Anticipated expiration: 2038-12-17
Also published as: AU2018391714A1; TW201927329A; TWI756500B; CN111655275B; WO2019120162A1

Abstract

The present invention relates to a method for treating type II diabetes. More particularly, the present invention relates to a method for synergistically lowering the blood glucose level of an individual having type II diabetes mellitus using a wild ginger flower plant extract and a hypoglycemic agent.

Description

Compositions, kits and methods for treating type II diabetes

Related application

The present application claims priority date of provisional application No. 62/599,822 filed on 2017, 12/18/35, by code 119(e) of U.S. 35, the entire contents of which are incorporated herein by reference.

Background

1. Field of the invention

The present invention relates to a method for treating hyperglycemia, in particular type II diabetes mellitus.

2. Background of the invention

Type ii diabetes or non-insulin dependent diabetes mellitus (NIDDM) is known to have a genetic element susceptible to external environmental factors. The underlying causes of type ii diabetes include defects in the beta cells that produce insulin; changes in muscle cells, adipocytes, and hepatocytes response to insulin; and abnormalities in the regulatory mechanisms responsible for controlling carbohydrate and lipid metabolism after ingestion of food. Modulation of insulin sensitivity is influenced by environmental factors and behaviors, mainly by a quiescent (sedentary) lifestyle and obesity. The cellular mechanisms that contribute to the regulation of insulin sensitivity in muscle and fat cells are rather complex and not yet fully understood. It is believed that altering insulin signaling pathways, increasing the amount of intracellular fat, and increasing the levels of free fatty acids and other adipose tissue products can affect insulin sensitivity.

Hyperglycemic state (hyperglycemic state), if not properly controlled or maintained, inevitably leads to complications (complications) including cardiovascular disease, visual impairment, various forms of neuropathy and cognitive impairment, stroke, and peripheral vascular disease. In addition to making major adjustments to the dietary nutrition and physical activity of an individual, common treatment regimens include the use of antihyperglycemic drugs and insulin. Since the disease is chronic and progressive, there is no treatment that reverses the disease process to date, and there remains a need in the art for an effective method and/or drug that can be used to treat type ii diabetes.

Disclosure of Invention

The following presents a simplified summary of the invention in order to provide a basic understanding to the reader. This summary is not an extensive overview of the invention and is not intended to identify key/critical elements of the invention or to delineate the scope of the invention. Its sole purpose is to present some concepts of the invention in a simplified conceptual form as a prelude to the more detailed description that is presented later.

The present invention relates to a drug which can be used alone or in combination with any hypoglycemic agent, and which is effective in lowering the blood glucose level of an individual with hyperglycemia (particularly an individual with type II diabetes).

Accordingly, one aspect of the present invention is directed to a method for treating a subject having type ii diabetes. The method comprises administering to the subject an effective amount of a flower of wild ginger (Hedychium coronarium Koenig) plant extract and a hypoglycemic agent, wherein the combination therapy synergistically lowers the blood glucose level of the subject to alleviate and/or ameliorate symptoms associated with type II diabetes.

According to an embodiment of the present invention, a method for preparing a wild ginger flower plant extract suitable for use in the treatment method of the present invention comprises:

(a) extracting an aerial portion of a flower of wild ginger (Hedychium coronarium Koenig) with a solvent to produce a first extract, wherein the solvent is (1) petroleum ether (petroleum ether), (2) n-hexane, (3) dichloromethane, (4) trichloromethane, (5) ethyl acetate, (6) acetone, or (7) ethanol at a concentration of 70 to 100% (v/v in water), or (8) any combination of (1) to (7);

(b) placing the first extract into a first ion exchange chromatography column;

(c) washing the first ion exchange chromatography column with a solution of water and ethanol in a volume ratio of 1:1 to 1: 9; and

(d) washing the first ion exchange chromatography column with an ethanol concentration of at least 70% to produce the extract of the flowers of wild ginger.

According to an embodiment of the present invention, the extract of the flowers of wild ginger prepared by the above method substantially has an HPLC profile as shown in fig. 1A.

According to an embodiment of the present invention, the extract of the flowers of wild ginger prepared by the aforementioned method substantially has an HPLC profile as shown in fig. 1B.

According to an embodiment of the present invention, the hypoglycemic agent may be selected from the group consisting of dipeptidyl peptidase-4 (DPP-4) inhibitors, insulin analogs, biguanides (biguanides), sulfonylureas (sulfonureas), Thiazolidinediones (TZDs), sodium-glucose cotransporter 2(SGLT2) inhibitors, alpha-glucosidase (alpha-glucosidase) inhibitors, glucagon-like peptide 1(glucagon-like peptide 1, GLP-1) receptor agonists, and combinations thereof.

According to an embodiment of the present invention, the DPP-4 inhibitor is gliptin (gliptin). Examples of suitable gliptins include, but are not limited to: sitagliptin (sitagliptin), vildagliptin (vildagliptin), saxagliptin (saxagliptin), linagliptin (linagliptin), gemagliptin (gemagliptin), alagliptin (anagliptin), trelagliptin (teneligliptin), alogliptin (alogliptin), trelagliptin (trelagliptin), dologliptin (dutogliptin), alogliptin (omarigliptin), orygliptin (omarigliptin), berberine (berberine), and lupeol (lupeol).

According to one embodiment of the invention, the method comprises administering to the subject an effective amount of the extract of the flowers of zingiber officinale and the DDP-4 inhibitor. The DDP-4 inhibitor is preferably sitagliptin.

According to an embodiment of the invention, the insulin analogue may be insulin glargine (glargine), deglutamic (degludec) or insulin detemir (detemir).

According to an embodiment of the present invention, the biguanide may be metformin (metformin), phenformin (phenformin) or buformin (buformin). According to a preferred embodiment of the present invention, the method comprises administering to the individual an effective amount of the extract of the flowers of zingiber officinale and the biguanide. The biguanide is preferably metformin.

According to an embodiment of the invention, the sulfonylurea is glibenclamide (glibenclamide), gliclazide (gliclazide), glimepiride (glimepiride) or glipizide (glipizide).

According to an embodiment of the invention, the thiazolidinedione is pioglitazone (pioglitazone), rosiglitazone (rosiglitazone), lobeglitazone (lobeglitazone), ciglitazone (ciglitazone), darglitazone (darglitazone), englitazone (englitazone), netglitazone (netoglitazone), rivoglitazone (rivoglitazone) or troglitazone (troglitazone).

According to an embodiment of the invention, the SGLT2 inhibitor is dapagliflozin (dapagliflozin), engeletzin (empagliflozin), canagliflozin (canagliflozin), egeletzin (Ipragliflozin), tofogliflozin (tofogliflozin), iseglucin (sergliflozin etabonate), remogliflozin (remogliflozin) or egeletzin (ertagliflozin).

According to an embodiment of the present invention, the α -glucosidase inhibitor may be acarbose (acarbose), miglitol (miglitol) or voglibose (voglibose).

According to an embodiment of the invention, the GLP-1 receptor agonist is liraglutide (liraglutide), exenatide (exenatide), albiglutide (albicidin) or LY 2189265.

According to an embodiment of the invention, the method comprises administering to the subject the extract of the plant zingiber officinale in combination with a biguanide and a DDP-4 inhibitor. The biguanide is preferably metformin, and the DDP-4 inhibitor is preferably sitagliptin.

According to another embodiment of the invention, the method of the invention comprises administering to the individual the extract of the plant zingiber officinale in combination with a biguanide and an SGLT2 inhibitor. The aforementioned biguanide is preferably metformin, and the SGLT2 inhibitor is preferably eggliflozin.

According to a preferred embodiment of the present invention, the wild ginger flower plant extract and the hypoglycemic agent may be administered individually to the individual by oral, intravenous, intramuscular, subcutaneous, transmucosal or intrarectal routes. In a preferred embodiment, the wild ginger flower extract and the hypoglycemic agent are administered to the individual orally. Accordingly, the extract of the wild ginger flower plant suitable for oral administration and the hypoglycemic agent may be formulated into a lozenge, pill, granule, powder, solution, suspension, syrup or capsule.

According to an embodiment of the present invention, about 0.1 to 1,000 mg/kg of the extract of the flowers of wild ginger and about a hypoglycemic agent are administered to the subject, respectively. Preferably, 1 to 500 mg/kg of the extract of the wild ginger flower plant and the hypoglycemic agent are administered to the subject, respectively.

The basic spirit and other objects of the present invention, as well as the technical means and aspects of the present invention will be readily apparent to those skilled in the art from the following detailed description of the embodiments.

Drawings

In order to make the aforementioned and other objects, features, advantages and embodiments of the invention more comprehensible, the following description is given:

FIG. 1A is an HPLC chromatogram of a wild ginger flower plant extract prepared according to example 1.1 of the present invention;

fig. 1B is an HPLC profile of a wild ginger flower plant extract prepared according to example 1.2 of the present invention;

FIG. 2 is a broken line graph of the blood glucose level of NIDDM mice obtained from the plant extract of flowers of Zingiber officinale (HC) and/or sitagliptin according to example 2 of the present invention;

FIG. 3 is a graph illustrating the results of the cumulative total blood glucose values under the curves of FIG. 2;

fig. 4 is a histogram illustrating the effect of any one of the zingiber officinale plant extract (HC), Sitagliptin (SITA) or Egyprost (ERTU) in combination with Metformin (MET) on glucose homeostasis, according to example 3.1 of the present invention, wherein a represents p <0.05 (compared to the control group);

FIG. 5 is a histogram illustrating the synergistic effect (synergistic effect) results of combined use of the extract of Zingiber officinale (HC), Metformin (MET) and Sitagliptin (SITA) in delaying the onset of diabetes compared to individual treatment results (1 day of fasting blood glucose increment +198mg/dL compared to 29 days of treatment with the extract of Zingiber officinale, 1 day of fasting blood glucose increment +222mg/dL compared to 29 days of treatment with sitagliptin/metformin), and synergistically delayed onset of diabetes (1 day of fasting blood glucose increment +3mg/dL compared to 1 day of treatment with 29 days of treatment with the extract of Zingiber officinale;

fig. 6 is a graph showing the results of the synergistic effect of the zingiber officinale plant extract (HC), Sitagliptin (SITA) and Metformin (MET) on glucose homeostasis according to example 3.2 of the present invention, wherein a represents p <0.05 (compared to the control group), b represents p <0.001 (compared to the HC group) and c represents p <0.01 (compared to the SITA + MET group);

FIG. 7 shows the results of the synergistic effect of the wild ginger flower plant extract (HC), eggliflozin ((ERTU) and Metformin (MET) on glucose homeostasis according to example 3.3 of the present invention, wherein a represents p <0.05 (compared to the control group), b represents p <0.05 (compared to the wild ginger flower plant extract) and c represents p <0.01 (compared to the ERTU + MET group), and

FIG. 8 illustrates the use of the extract of the flowers of wild ginger (HC) of example 4 of the present invention as an additional agent to metformin for the treatment of type II diabetics, wherein a represents p <0.05 (compared to MET).

Description of the invention

In order to make the description of the invention more complete and complete, the following description is given for illustrative purposes with respect to the implementation aspects and specific embodiments of the invention; it is not intended to be the only form in which the embodiments of the invention may be practiced or utilized. The embodiments are intended to cover the features of the various embodiments as well as the method steps and sequences for constructing and operating the embodiments. However, other embodiments may be utilized to achieve the same or equivalent functions and step sequences.

I. Definition of

For ease of description, specific terms recited in the specification, examples, and appended claims are systematically described herein. Unless defined otherwise herein, 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.

As used herein, the singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise. As used herein, "about" (about) generally refers to within 10%, 5%, 1%, or 0.5% of a given value or range. Alternatively, the term "about" means that the actual value falls within the acceptable standard error of the mean, subject to consideration by those of ordinary skill in the art to which the invention pertains. Except in the experimental examples, or where otherwise expressly indicated, it is to be understood that all ranges, amounts, values and percentages herein used (for example to describe amounts of materials, length of time, temperature, operating conditions, quantitative ratios or angles of reflection) are to be modified by "about". Accordingly, unless indicated to the contrary, the numerical parameters set forth in the specification and attached claims are approximations that may vary depending upon the desired properties sought to be obtained. At the very least, these numerical parameters are to be understood as meaning the number of significant digits recited and the number of significant digits that result from applying ordinary rounding techniques.

The "wild ginger flower" (Hedychium coronarium Koenig) used in the present invention is a monocotyledonous plant of the order Zingiberaceae (Zingiberales), the family Zingiberaceae (Zingiberaceae), the genus Iris (Hedychium). The plant is native to India, Malaysia and Himalayan mountains and is usually grown in low-altitude areas. In Taiwan, its traces can be found in Taibei, Xinzhu, Taizhong, Gao-Xiong and Yandong mountainous areas, fields and at the edge of ditches. The underground part of the plant comprises rhizomes (shaped like ginger); while the aerial parts include leaves and pseudostems (pseudostems) formed by leaf sheaths (leaf sheaths). The leaves are spear-shaped, 40 cm long and 7 cm wide, and have smooth surface and short villus on the back. The flower Lip (Lip) and petals are white and fragrant. The wild ginger flower plant can reach 1-2 m height, is a cluster, and is suitable for pot culture and flower arrangement. The tender shoots and roots are edible. The flowers of wild ginger are also called butterfly ginger (butterfly ginger), butterfly flower (butterfly lily), ginger orchid (ginger flower), white butterfly flower (white butterfly lily), Gandasuli (popular names of places such as indonesia, malaysia, etc.), and Kamia (popular names of places such as philippines, etc.).

The term "treatment" or "treating" as used herein is intended to mean obtaining a desired pharmacological and/or physiological effect, such as lowering the blood glucose level of an individual with hyperglycemia. The aforementioned effects may be prophylactic in terms of total or partial prevention of the disease or symptoms thereof; and/or the foregoing effects may be therapeutic in terms of treating the disease, in whole or in part, and/or due to adverse effects of the disease. "treating" or "treatment" as used herein includes, but is not limited to, prophylactic (e.g., prophylactic), curative or palliative treatment of a disease in a mammal, particularly a human; and comprises: (1) prophylactic (e.g., prophylactic), curative or palliative treatment of a disease or disorder (e.g., diabetes or symptoms associated therewith) that may be predisposed to the disease but has not yet been diagnosed as having the disease; (2) inhibiting a disease (e.g., by promoting proliferation of insulin-producing beta cells, or inhibiting apoptosis of such cells); or (3) ameliorating a disease (e.g., reducing symptoms associated with the disease).

The terms "administration," "administering," or "administration" (administered, or administration) are used interchangeably herein to refer to a mode of delivery, including, but not limited to: an agent (e.g., a compound or composition) of the invention is administered orally, intravenously, intramuscularly, intraperitoneally, intraarterially, intracranially, transmucosally (e.g., by inhalation and intranasally), or subcutaneously. In a preferred embodiment, the extract of the plant of the flowers of wild ginger of the present invention can be formulated into a composition suitable for oral administration.

As used herein, an "effective amount" refers to an amount effective to achieve the desired result in treating a disease caused by hyperglycemia in a particular desired time. For example, in the treatment of diabetes, an agent (i.e., a compound of the invention) that is effective to reduce, prevent, delay, inhibit or arrest any symptom associated with diabetes. An effective amount of the agent does not necessarily cure the disease or condition but provides treatment for a disease or condition, such as delaying the onset of the disease or condition, limiting or arresting the onset of the disease or condition, or ameliorating the symptoms of the disease or condition. The specific effective amount or sufficient amount may vary depending on various factors, such as the particular condition to be treated, the physiological condition of the patient (e.g., the weight, age, or sex of the patient), the type of mammal or animal being treated, the time of treatment, the nature of concurrent therapy sessions, if any, and the particular formulation employed. For example, an effective amount may be expressed as the total mass of active pharmaceutical agent per day (e.g., in grams, milligrams, or micrograms); or expressed as the weight of active agent per kilogram of body weight. An effective amount may be divided into one, two or more doses in a suitable form for administration in one, two or more doses over a specified period of time.

The terms "individual", "subject" or "patient", "patient" are used interchangeably herein and mean a mammal (including a human) capable of being treated with a compound of the invention. By "mammal" (mmal) is meant all members of the class of mammals (class), including humans, primates, domesticated and livestock animals (e.g., rabbits, pigs, sheep, cattle) and zoo captive animals, animals for sports, or pets; and rodents (e.g., mice and rats). Furthermore, unless gender is explicitly indicated, the terms "individual" or "subject" or "patient" include both males (males) and females (females). Thus, the terms "individual", "subject", "patient" or "patient" include any mammal that would benefit from the treatment methods of the present invention. Examples of "individual", "subject", "patient" or "patient" include, but are not limited to: humans, rats, mice, guinea pigs, monkeys, pigs, goats, cattle, horses, dogs, cats, birds and birds. In a preferred embodiment, the subject is a human.

The term "synergistic reduction" (e.g., lowering of blood glucose level) as used in the present invention is an effect achieved by the combination of the wild ginger flower plant extract of the present invention and the hypoglycemic agent, and can be superior to the sum of the effects achieved by the use of the wild ginger flower plant extract of the present invention or the hypoglycemic agent, respectively. Advantageously, such synergy (synergy) may provide better efficacy at the same dosage.

The term "symptoms associated with type II diabetes mellitus" as used herein is meant to include acute and/or chronic symptoms including at least hyperglycemia, insulin resistance, increased thirst and/or hunger, frequent urination, abnormal weight loss, fatigue, and slow healing of sores.

The term "excipient", as used herein, refers to any inert substance (e.g., powder or liquid) that can form a carrier (vehicle) for an active agent. Excipients are generally safe, non-toxic, and in a broad sense may also include any material known in the pharmaceutical industry for the preparation of pharmaceutical compositions, such as fillers, diluents, agglutinating agents, binders, lubricants, glidants, stabilizers, coloring agents, wetting agents, disintegrants, and the like.

Treatment of type II diabetes

In general, the present invention relates to the discovery that a plant extract of flowers of wild ginger can act synergistically with known hypoglycemic agents (e.g., DDP-4 inhibitors) to reduce fasting blood glucose levels (fasting blood glucose levels) in a diabetic individual, particularly in individuals with type II diabetes. Accordingly, the extract of the wild ginger flower plant can be used as an additional drug to any known hypoglycemic agent for the treatment of type II diabetes.

In this respect, a particular aspect of the invention relates to a method of treating an individual suffering from type II diabetes. The method comprises the following steps: administering to the subject an effective amount of a wild ginger flower plant extract and a hypoglycemic agent, wherein the combination therapy synergistically lowers the blood glucose level of the subject.

The wild ginger flower plant extract of the present invention, particularly the ethanol extract obtained from the aerial parts of wild ginger flowers, is preferably prepared according to the method described in U.S. patent application No. 15/359,555, or U.S. patent publication No. 9,023,407. These inventions are incorporated as part of the present invention by reference.

Preferably, the plant extract of the present invention is prepared by a method comprising the steps of:

(a) extracting an aerial portion of a flower of zingiber officinale (Hedychium coronarium Koenig) with a first solvent to produce a first extract, wherein the first solvent is (1) petroleum ether (petroleum ether), (2) n-hexane, (3) dichloromethane, (4) chloroform, (5) ethyl acetate, (6) acetone, or (7) ethanol at a concentration of 70 to 100% (v/v in water), or (8) any combination of (1) to (7);

(b) placing the first extract into a first ion exchange chromatography column;

(c) in a volume ratio of 1:1 to 1:9 washing the first ion exchange chromatography column with an ethanol solution; and

(d) washing the first ion exchange chromatography column with an ethanol concentration of at least 70% to prepare the plant extract.

According to an embodiment of the present invention, the aerial parts (e.g. pseudostems and/or leaves) of the flowers of wild ginger are extracted with 95% ethanol (v/v in water).

The prepared wild ginger flower plant extract may be further subjected to at least one liquid chromatography (e.g., high performance liquid chromatography (hereinafter referred to as HPLC) treatment, according to a preferred embodiment of the present invention, the wild ginger flower plant extract is subjected to one HPLC treatment, according to another preferred embodiment of the present invention, the wild ginger flower plant extract is subjected to at least two HPLC treatments for subsequent purification, according to a preferred embodiment of the present invention, the wild ginger flower plant extract of the present invention is substantially characterized by having an HPLC profile as depicted in fig. 1A, according to another preferred embodiment of the present invention, the wild ginger flower plant extract of the present invention is substantially characterized by having an HPLC profile as depicted in fig. 1B.

According to a preferred embodiment of the present invention, the extract of the flowers of wild ginger is administered with one or more hypoglycemic agents, which achieve a synergistic effect in lowering blood glucose levels in an individual. Any known hypoglycemic agent may be used. Preferably, the hypoglycemic agent is a dipeptidyl peptidase-4 (DPP-4) inhibitor, insulin, an insulin analog, a biguanide, a sulfonylurea, a Thiazolidinedione (TZD), a sodium-glucose cotransporter 2(SGLT2) inhibitor, an alpha-glucosidase (alpha-glucosidase) inhibitor, a glucagon-like peptide 1(GLP-1) receptor agonist, or a combination thereof.

The DPP-4 inhibitor is gliptin (gliptin), suitable examples include, but are not limited to, sitagliptin (sitagliptin), vildagliptin (vildagliptin), saxagliptin (saxagliptin), linagliptin (linagliptin), gemagliptin (gemagliptin), alagliptin (anagliptin), trelagliptin (teneligliptin), alogliptin (alogliptin), trelagliptin (trelagliptin), dologliptin (dutogliptin), alogliptin (omarigliptin), minilin (berberine), and lupeol (lupeol). Preferably, the DPP-4 inhibitor is sitagliptin.

The term "insulin" is used herein to refer to a purified, synthetic and/or biotechnologically derived product that is identical or similar in structure, use and expected effect to naturally occurring insulin and that has value in the treatment of diabetes. For example, insulin can be recovered directly from pancreatic tissue of a mammal, such as from the pancreatic glands of livestock (e.g., swine). Alternatively, insulin may also be prepared by recombinant techniques.

Examples of insulin analogs include, but are not limited to, insulin glargine, insulin deglutamide, and insulin detemir.

Suitable examples of biguanides include, but are not limited to, metformin, phenformin, and buformin.

Suitable examples of sulfonylureas include, but are not limited to, glibenclamide (glibenclamide), gliclazide (gliclazide), glimepiride (glimepiride), or glipizide (glipizide).

Suitable examples of TZDs include pioglitazone (pioglitazone), rosiglitazone (rosiglitazone), lobeglitazone (lobeglitazone), ciglitazone (ciglitazone), darglitazone (darglitazone), englitazone (englitazone), netoglitazone (netoglitazone), rivoglitazone (rivoglitazone), or troglitazone (troglitazone), without being limited thereto.

Examples of SGLT2 inhibitors include dapagliflozin (dapagliflozin), englobin (empagliflozin), canagliflozin (canagliflozin), Ipragliflozin (Ipragliflozin), tofogliflozin (tofogliflozin), iseggliflozin (servlifzin etabonate), remogliflozin (remogliflozin etabonate), or eggliflozin (ertagliflozin), but are not limited thereto.

Suitable examples of alpha-glycosidases include acarbose (acarbose), miglitol (miglitol) or voglibose (voglibose), although the invention is not limited thereto.

Suitable examples of GLP-1 receptor agonists include liraglutide (liraglutide), exenatide (exenatide), albiglutide (albicidin), or LY2189265, although the present invention is not limited thereto.

According to some embodiments of the invention, a wild ginger flower plant extract is administered with a DDP-4 inhibitor (e.g., sitagliptin), wherein the combination therapy synergistically lowers blood glucose levels.

According to other embodiments of the present invention, a combination of a extract of a plant of Zingiber officinale and a biguanide (e.g., metformin) is administered to an individual in need thereof, wherein the combination treatment synergistically lowers blood glucose levels.

According to another embodiment of the present invention, a combination of a wild ginger plant extract and an SGLT2 inhibitor (e.g., eggliflozin) is administered to an individual in need thereof, wherein the combination treatment synergistically lowers blood glucose levels.

According to yet another embodiment of the present invention, a combination of a extract of the plant Zingiber officinale with a biguanide (e.g., metformin) and a DDP-4 inhibitor (e.g., sitagliptin) is administered to an individual in need thereof, and such combination treatment synergistically lowers blood glucose levels.

In accordance with yet another embodiment of the present invention, a combination of a extract of a plant of Zingiber officinale and a biguanide (e.g., metformin), and an SGLT2 inhibitor (e.g., eggliflozin) is administered to an individual in need thereof, wherein the combination treatment synergistically lowers blood glucose levels.

According to embodiments of the present invention, 0.1 to 2,000 mg per day of the extract of the wild ginger flower and the hypoglycemic agent, such as 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1,2, 3,4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 74, 72, 76, 80, 84, 80, 84, 83, 85, 83, 85, 83, 82, 83, 82, 83, 82, 23, 3,4, 95. 96, 97, 98, 99, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550, 560, 570, 580, 600, 610, 620, 630, 640, 650, 660, 670, 680, 690, 700, 710, 720, 730, 740, 750, 760, 770, 780, 790, 800, 810, 820, 830, 840, 850, 860, 870, 880, 890, 900, 910, 930, 940, 950, 960, 970, 980, 990, 1,000, 1,100, 1,200, 1,300, 1,400, 1,500, 1,600, 1,700, 900, 1,800, 900, 1,000, and 2,000 milligrams; preferably about 0.5 to 800 mg per day of the wild ginger flower plant extract and the hypoglycemic agent, e.g. 0.5, 0.6, 0.7, 0.8, 0.9, 1,2, 3,4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 88, 90, 91, 92, 90, 96, 97, 96, 99, 95, 98, 99, 98, 100, 98, 170. 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550, 560, 570, 580, 590, 600, 610, 620, 630, 640, 650, 660, 670, 680, 690, 700, 710, 720, 730, 740, 750, 760, 770, 780, 790 and 800 milligrams; more preferably about 1 to 500 mg per day of the wild ginger flower plant extract and the hypoglycemic agent, e.g. 1,2, 3,4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 120, 100, 180, 170, 180, 150, 100, 180, 150, 180, 150, 100, 150, 180, 170, 180, 150, 230. 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, and 500 milligrams. According to a particular embodiment of the invention, the extract of the wild ginger flower plant and the hypoglycemic agent (i.e., sitagliptin) are administered to a human subject in amounts of about 11 mg per kg and about 3.5 mg per kg, respectively.

According to embodiments of the present invention, the wild ginger plant extract or hypoglycemic agent, respectively, may be administered by any suitable route, including, but not limited to, oral, intravenous, intramuscular, intraperitoneal, intraarterial, intracranial, and subcutaneous routes. In preferred embodiments, an effective amount of the extract of the flowers of wild ginger and a hypoglycemic agent are each administered orally to an individual in need thereof.

The invention also includes pharmaceutical compositions for treating type II diabetes. The composition comprises a wild ginger flower plant extract, a hypoglycemic agent and a pharmaceutically acceptable excipient. Any known hypoglycemic agent may be used in the compositions of the present invention. Preferably, the hypoglycemic agent is a dipeptidyl peptidase-4 (DPP-4) inhibitor, insulin analog, biguanide, sulfonylurea, Thiazolidinedione (TZD), sodium-glucose cotransporter 2(SGLT2) inhibitor, alpha-glucosidase (alpha-glucosidase) inhibitor, glucagon-like peptide 1(GLP-1) receptor agonist, or a combination thereof.

For the preparation of a pharmaceutical composition, the extract of the flowers of wild ginger is mixed with a hypoglycemic agent and suitable excipients and formulated in a dosage form suitable for oral, intravenous, intramuscular, subcutaneous, intraarterial, intracranial, transmucosal (e.g. inhalation, buccal or intranasal) or rectal administration. Suitable excipients are well known in the art and are described, for example, in the handbook of Pharmaceutical excipients (Kibbe (ed.),3rd Edition (2000), American society of pharmacists, Washington, D.C.), and Remington's Pharmaceutical Sciences (Gennaro (ed.),20th Edition (2000), Mack Publishing Inc., Easton, Bin), the disclosures of which are incorporated herein by reference. For example, suitable excipients include, but are not limited to, starches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrating agents, wetting agents, lubricants (which occur repeatedly), emulsifying agents, coloring agents, deforming agents, coating agents, sweetening agents, flavoring agents, preservatives, plasticizers, gelling agents, thickening agents, hardening agents, flash-setting agents, suspending agents, surfactants, humectants, carriers, stabilizers, antioxidants, and combinations thereof.

The pharmaceutical compositions are generally prepared in dosage forms suitable for administration to a subject by any desired route. Those of ordinary skill in the art are familiar with the various dosage forms suitable for use in the present invention. The most suitable route in any given case will depend on the nature of the disease being treated and/or managed and its severity. For example, pharmaceutical compositions can be formulated for oral, intravenous, intramuscular, intraperitoneal, intraarterial, intracranial, transmucosal (e.g., inhalation, buccal, and intranasal), or subcutaneous administration. The pharmaceutical composition is preferably formulated for oral administration. For example, dosage forms of pharmaceutical compositions suitable for oral administration include: lozenges, pills, granules, powders, solutions, suspensions, syrups or capsules. As to the method of preparing solid dosage forms such as lozenges, pills, granules or powders, pharmaceutically acceptable carriers (e.g., excipients, binders or disintegrants, etc.) may be used. Solid dosage forms suitable for oral administration may optionally be scored or prepared with coatings and shells, such as enteric coatings and coatings to modify the rate of release. In addition, any solid dosage form may be enclosed in soft and hard gelatin capsules using any excipient known in the art.

The wild ginger flower plant extract and the hypoglycemic agent may also be formulated in a liquid dosage form suitable for oral administration. Suitable formulations include emulsifiers, solutions, suspensions or syrups which can be prepared by using diluents commonly used in the art, such as water, glycerol, alcohols, vegetable oils, etc. Liquid formulations may optionally include adjuvants such as wetting agents, emulsifying agents, suspending agents, sweetening agents, flavoring agents, coloring agents and preserving agents. The liquid preparation can also be filled in soft gelatin capsule. For example, the liquid may comprise a solution, suspension, emulsion, precipitate or any other desired liquid medium carrying the extract of the flowers of the wild ginger plant and the hypoglycemic agent. The liquid may be designed to improve the solubility of the wild ginger flower plant extract and the hypoglycemic agent upon release, or may be designed to form an emulsified or dispersed phase containing the drug upon release. Examples of the foregoing techniques are generally known in the art. Soft gelatin capsules may be provided with a functional coating, e.g., a coating that delays release of the drug, as desired.

In the case of injectable administration (or parenteral administration), the extract of the flowers of wild ginger and the hypoglycemic agent may be prepared in an injectable form for intravenous, subcutaneous or intramuscular administration. The aforementioned injection can be prepared by dissolving the extract of the wild ginger flower plant and the hypoglycemic agent in an aqueous solution such as physiological saline or in a non-aqueous solution composed of organic esters such as propylene glycol, polyethylene glycol or vegetable oil such as sesame oil.

In the case of transdermal administration, dosage forms such as ointments or creams may be used, for example. The ointment can be prepared by mixing the extract of the wild ginger flower and the hypoglycemic agent with fat or oil and the like; the cream can be prepared by mixing the extract of the wild ginger flower plant, the hypoglycemic agent and the emulsifier. Transdermal formulations may also be prepared in liquid form or in powder formulations. In the liquid formulation, water, saline solution, phosphate buffer, acetate buffer, and the like may be used as a base; surfactants, antioxidants, stabilizers, preservatives or viscosity increasing agents may also be present. In the case of powder formulations, water-absorbing materials such as water-soluble polyacrylates, cellulose oligoesters, polyethylene glycols, polyvinylpyrrolidone, amylases and the like; and non-water absorbing materials such as cellulose, starch, gums, vegetable oils, or cross-linked polymers. In addition, antioxidants, colorants and preservatives may also be added to the powder formulation. Liquid or powder formulations may be administered by use of a spray device.

In the case of rectal administration, this is in the form of a plug of soft gelatin capsules.

In the case of inhalation via oronasal breathing, a solution or suspension containing the extract of the flowers of wild ginger and a hypoglycemic agent, together with the pharmaceutical excipients generally used for this purpose, can be inhaled by means of an inhalation aerosol spray. Alternatively, the powder form of the extract of the wild ginger flower and the hypoglycemic agent may be administered by an inhaler which allows the powder to be in direct contact with the lungs. Pharmaceutically acceptable carriers (e.g., isotonic agents, preservatives, dispersants or stabilizers) may optionally be added to the aforementioned formulations. In addition, these formulations may be sterilized by filtration or by exposure to high temperature or radiation, as desired.

Generally, a single dose or multiple doses of a pharmaceutical composition comprising a wild ginger flower plant extract and a hypoglycemic agent are administered to an individual 2 times, 3 times, 4 times, 5 times, 6 times or more daily. Alternatively, the administration may be once every two days, three days, four days, five days, six days, or more. In a preferred embodiment, the pharmaceutical composition is administered once daily. In other embodiments, the pharmaceutical composition is administered twice daily.

Another aspect of the invention relates to a kit that can be used to treat type ii diabetes. The kit at least comprises a first container, wherein the wild ginger flower plant extract is contained in the first container; a second container comprising a hypoglycemic agent; and an illustration providing an instruction for a user to administer the extract of the flowers of wild ginger and the hypoglycemic agent to treat type II diabetes.

According to a preferred embodiment of the invention, the kit comprises two containers, wherein the first container comprises a dosage form of a wild ginger plant extract and the second container comprises a dosage form of a DDP-4 inhibitor (e.g., sitagliptin), a biguanide, an SGLT2 inhibitor or any combination of the foregoing; the kit also includes a legend to provide instructions for use of the kit. The legend may be in the form of a booklet, tape, CD, VCD or DVD.

The wild ginger flower plant extract and the hypoglycemic agent of the kit of the present invention may be individually formulated in dosage forms suitable for administration to a subject by any desired route. Those of ordinary skill in the art are familiar with the various dosage forms suitable for use in the present invention. The most suitable route in any given case will depend on the nature of the disease being treated and/or managed and its severity. For example, the extract of the wild ginger flower of the present invention and the hypoglycemic agent may be formulated for oral, intravenous, intramuscular, intraperitoneal, intraarterial, intracranial, transmucosal (e.g., inhalation, buccal, or intranasal) or subcutaneous administration. Preferably, the extract of the wild ginger flower plant of the present invention and the hypoglycemic agent may be orally administered separately.

The present invention will now be described in further detail with reference to the following examples. However, it should be understood that the present invention is not limited to the specific examples.

Examples

Materials and methods

Laboratory animal

The experiment mainly uses Non-insulin dependent diabetes mellitus (Non-insulin dependent diabetes mellitus, NIDDM) db/db male mice (BKS. Cg-Dock 7)^m+/+Lepr^db/JNarl). Experimental animals were provided mainly by The Jackson memorial Laboratory (The Jackson Laboratory, Bar Harbor, maine, usa). These experimental mice exhibited hyperinsulinemia, hyperglycemia, and islet atrophy. Mice 9-13 weeks old were used mainly for this experiment.

All mice were housed in an animal house with temperature controlled at 20 to 24 ℃ and humidity of 50 to 80%) and with 12 hours/12 hours on a day/night cycle (light on time set at 7:00a.m.), with food and water ad libitum. According to the following guidelines: animal experiments were conducted in AAALAC approved Laboratory animal rooms (Europe group Pan Biochemist Co., Ltd.) under relevant regulations described in Guide for the Care and Use of Laboratory Animals: EighthEdition, National academy Press, Washington D.C., 2011.

NIDDM mice and treatment

NIDDM mice were acclimated for at least five days and then treated in groups when the mean blood glucose values were greater than 180mg/dl after overnight fasting (day 0). As part of example 2 described below, the test compound (i.e., the wild ginger plant extract (administered orally at a dose of 40 mg per kg on days 1 to 14; administered orally at a dose of 80mg per kg on days 14 to 28), sitagliptin (administered at a dose of 40 mg per kg) or a combination of the wild ginger plant extract and sitagliptin) was fed to each mouse at the indicated dose beginning on day 1 by oral gavage and continued for 27 days until day 28. In example 3 described later, test compounds (i.e., the extract of the wild ginger flower alone (orally administered at a dose of 40 mg per kg on days 1 to 28), or the extract of the wild ginger flower together with metformin (orally administered at a dose of 150 mg per kg), sitagliptin (orally administered at a dose of 40 mg per kg) or eggliflozin (orally administered at a dose of 10 mg per kg) were fed to each mouse at the prescribed dose starting on day 1 by oral gavage and continued for 27 days until day 28.

The values of blood glucose, insulin, glycated hemoglobin (HbA1c) were measured on the indicated days after overnight fasting. The effect of each treatment on glucose homeostasis was then evaluated in an Oral Glucose Tolerance Test (OGTT). The body weight of each test mouse was also measured during the experiment.

Oral Glucose Tolerance Test (OGTT)

The foregoing db/db mice were randomly divided into four groups: (1) a vector control group; (2) HC plant extract (80 mg per kg); (3) sitagliptin (40 mg per kg) group; and (4) HC plant extract (80 mg per kg) + sitagliptin (40 mg per kg).

Mice were fasted for 5 hours before receiving the indicated treatments individually. After 30 minutes, each mouse was orally gavaged (p.o.) with 10mL of glucose solution (concentration of 2 grams of glucose per kilogram of water) per kg. Blood was drawn from each mouse at intervals of 0, 30, 60, 90 and 120 minutes, and the blood glucose value (mg/dl) was measured.

Statistics of

Results are expressed as mean ± Standard Error of Mean (SEM). Unpaired t-tests (unpaired's t-test) or one-way ANOVA (one-way ANOVA) were used to statistically compare the differences between the groups of substance and vehicle treatments. A significant difference was identified if P <0.05 compared to the vector control group.

Example 1: preparation of wild ginger flower plant extract

Generally, an extract of aerial parts of flowers of wild ginger (Hedychium coronarium Koenig, this example is abbreviated as HC in the following paragraphs) was prepared according to the procedure described in U.S. Pat. No. 9,023,407. These disclosures are incorporated as part of the present invention by reference.

1.1 preparation of ethanol-extracted HC plant extract

The leaves and pseudostems were HC-dried and extracted twice with 15 volumes of 95% ethanol (EtOH) (volume ratio 1: 15). Concentrating the 95% ethanol extract and passing it through

Pipe string, sequentially using H₂O and 95% EtOH (2: 8) (F1), 95% EtOH (F)F2) 95% EtOH and ethyl acetate (EtOAc) (1: 1) (F3) washing. The F2 fraction (or "HC plant extract") was collected for subsequent glucose tolerance experiments.

Fig. 1A then presents the aforementioned F2 fraction of a characteristic High Performance Liquid Chromatography (HPLC) profile, wherein HPLC is performed under the following conditions: at 0 min mobile phase: 66% MeOH/34% H₂O; at 60 minutes mobile phase: 100% MeOH/0% H₂O; at 76 minutes mobile phase: 100% MeOH/0% H₂O; at 78 minutes mobile phase: 66% MeOH/34% H₂O; and mobile phase at 86 minutes: 66% MeOH/34% H₂O; flow rate: 1.0 ml per minute; and detecting wavelength: 254 nm.

1.2 preparation of Water-extracted HC plant extracts

The leaves and pseudostems were HC dried and extracted twice with 15 volumes of 50% ethanol (volume ratio 1:15) (w: v ═ 1: 15). Concentrating the above 50% ethanol extract and passing it through

Pipe string, sequentially using H₂O and 30% ethanol to prepare HC plant extract.

The resulting HC plant extracts were subjected to HPLC analysis using a linear gradient of Acetonitrile (ACN) and water containing 0.1% trifluoroacetic acid (TFA), and the characteristic high performance liquid chromatography profile exhibited is presented in fig. 1B. Wherein the HPLC analysis was performed under the following conditions: at 0 min mobile phase: 10% ACN (with 0.05% TFA)/90% H₂O (with 0.05% TFA); at 80 minutes mobile phase: 17% ACN (with 0.05% TFA)/83% H₂O (with 0.05% TFA); at 90 minutes of mobile phase: 100% ACN (with 0.05% TFA)/0% H₂O (with 0.05% TFA); at 100 minutes of mobile phase: 100% ACN (with 0.05% TFA)/0% H₂O (with 0.05% TFA); at 102 minutes of mobile phase: 10% ACN (with 0.05% TFA)/90% H₂O (with 0.05% TFA); and at 110 minutes mobile phase: 10% ACN (with 0.05% TFA)/90% H₂O (with 0.05% TFA); flow rate: 1.0 ml per minute; and detecting wavelength: 254 nm.

Example 2: blood glucose level and glucose tolerance test of diabetic mice treated with HC plant extract and DDP-4 inhibitor

In this example, NIDDM mice, which are slightly deficient in the insulin signaling cascade (cascade) at birth, leading to insulin resistance and progressing to the diabetic phenotype, were used to assess the degree of tolerance of the HC plant extract and/or sitagliptin (i.e., DDP-4 inhibitor) of example 1 to blood glucose and glucose. The experimental animals (mice) were treated in the manner described in the materials and methods section.

As expected, HC plant extract (80 mg per kg) and sitagliptin (40 mg per kg) resulted in a decrease in fasting plasma glucose values, respectively, in NIDDM mice; however, more surprisingly, a synergistic decrease in fasting glucose values was found 30 minutes after administration of glucose when HC plant extract and sitagliptin were administered simultaneously compared to NIDDM mice treated with HC plant extract or sitagliptin only (fig. 2).

The data at the 30 minute time point after glucose treatment was further analyzed as shown in fig. 3. HC plant extract alone (80 mg per kg) reduced the total blood glucose value by about 10.5% and sitagliptin (40 mg per kg) reduced the total blood glucose value by about 11.3%, indicating that sitagliptin was slightly more effective than HC plant extract. However, when HC plant extract was administered with sitagliptin, a total blood glucose value was found to be reduced by 76.7%.

Example 3: one of DDP-4 inhibitor, biguanide or SGLT2 inhibitor and HC plant extract are used in combination for treating blood glucose level and diabetes of NIDDM mouse

In this example, the combined effect of HC plant extract (80 mg per kg) and at least one of the following hypoglycemic agents on fasting plasma glucose and glycated hemoglobin (HbA1c) was further investigated in NIDDM mice. The hypoglycemic agent is metformin (150 mg per kg), sitagliptin (40 mg per kg), egagliflozin (10 mg per kg), or a combination thereof.

Briefly, test compounds are administered orally to NIDDM mice (about 9 weeks of age) for 4 weeks, and blood glucose values are measured initially (i.e., day 1) and at the end of the 4-week treatment period (i.e., day 29). Furthermore, an Oral Glucose Tolerance Test (OGTT) was performed at the end of the 4-week treatment period (i.e., day 29) to assess the effect of each treatment on glucose homeostasis. Area under the curve (AUC) of the incremental glucose is estimated by integrating the glucose values above the baseline according to the trapezoidal rule. One-way anova was performed to assess significance between each treatment.

Combination treatment of metformin and any of HC plant extract, DDP-4 inhibitor or SGLT2 inhibitor to significantly improve glucose tolerance

Referring to fig. 4, it is shown that blood glucose values are reduced by treatment of any one of HC plant extract (80 mg per kg), sitagliptin (40 mg per kg) or egagliflozin (10 mg per kg) in combination with metformin (150 mg per kg). The three combinations are effective in lowering blood glucose level. The combination of HC plant extract and metformin reduced the area under the incremental glucose curve by about 59%, while the combination of metformin and sitagliptin (40 mg per kg) or the combination of metformin and egagliflozin (10 mg per kg) reduced the area under the incremental glucose curve by about 56% and 39%, respectively, compared to the control group.

Accordingly, the HC plant extract of the present invention more effectively promotes the effect of metformin on lowering blood glucose level than known drugs such as DPP-4 inhibitors and SGLT2 inhibitors.

3.2HC plant extract, biguanide and DDP-4 inhibitor synergistically delay the progression of diabetes onset

Referring first to fig. 5, fasting blood glucose values are shown for co-treatment with HC plant extract (80 mg per kg), metformin (150 mg per kg) and sitagliptin (40 mg per kg). As shown, administration of HC plant extract alone was more effective in lowering blood glucose levels at day 29 (D29) than in combination with metformin and sitagliptin, respectively. Unexpectedly, a synergistic reduction in blood glucose to background blood glucose values can be achieved by including HC plant extracts in the metformin and sitagliptin combination (day 1, D1).

In addition, the incremental blood glucose results shown in fig. 6 demonstrate the efficacy of the combination of HC plant extract, metformin, and sitagliptin in producing a synergistic reduction in total blood glucose (compared to blood glucose values of the control group), and by about 123%.

3.3HC plant extract, biguanide and SGLT2 inhibitor synergistically delay the progression of diabetes

Referring to fig. 7, blood glucose results are shown for HC plant extract (80 mg per kg), metformin (150 mg per kg) and eggliflozin (10 mg per kg). As shown, HC plant extract alone, and the combination of metformin and eggliflozin were effective in lowering blood glucose values compared to the control group. The unexpected result was that inclusion of HC plant extract in the combination of metformin and eggliflozin synergistically reduced the area under the incremental glucose curve by up to 90.8%, compared to only 26.7% HC plant extract alone and only 38.9% metformin + eggliflozin alone.

In summary, the data in this example show that the HC plant extracts of the invention can synergistically improve glycemic control, including delaying the progression of diabetes, in NIDDM mice.

Example 4: HC plant extract as additional therapeutic agent for metformin for second type diabetic patients

In this example, the HC plant extract of the invention was used as an additional therapeutic agent for metformin in a clinical trial for treating patients with type ii diabetes.

Clinical trials were conducted on 20-70 year old patients diagnosed with type ii diabetes in taiwan, each patient receiving prescribed therapy. Oral glucose tolerance tests were performed and blood samples were collected at the indicated times as described in the materials and methods section.

Open-label tests (open-label), single-center randomized tests were performed through the three-military general hospital inland areas. Six to eight patients meeting the inclusion and/or exclusion criteria were enrolled with informed consent.

Inclusion condition (inclusion criterion):

1. male or female between 20 and 70 years of age.

2. Was diagnosed as type II diabetes (according to WHO 1999).

3. After 850 mg of metformin monotherapy per day for at least three months, no effective improvement in type II diabetes is seen.

HbA1c values between 7.0% and 9.0% (inclusive).

BMI of up to 35kg/m²。

6. Subjects were willing and able to follow the study procedure and sign informed consent.

Exclusion conditions (exclusion criterion):

1. known or suspected to be allergic to any component of the product of the study.

2. Fertility potential is maintained during the test period, either during pregnancy, lactation or prior to menopause, but at least two forms of contraception are not employed (at least one of which must be a contraceptive disorder).

And (4) note marking:

the following forms are included and acceptable:

1. contraceptive methods using oral, injectable or implantable hormones have been established.

2. An intrauterine device (IUD) or an intrauterine device containing lutein (IUS) is placed.

3. A disorder contraception method: condom or closure cap (septum or cervical/vaginal vault cap) coated with spermicidal foam/gel/coating/cream/suppository.

3. Another clinical trial was enrolled and received a study drug four weeks prior to the trial.

4. Impaired liver function is defined as alanine Aminotransferase (ALT), aspartate Aminotransferase (AST), or alkaline phosphatase (ALP) being at least 2.5 times the upper reference limit.

Impaired renal function 5 is defined as a male serum-creatinine of at least 1.3mg/dL (at least 115. mu. mol/L) and a female serum-creatinine of at least 1.2mg/dL (at least 106. mu. mol/L).

6. Any uncontrolled disease or any history of disease (e.g., hyperthyroidism) that may be detrimental to the subject upon entry into the trial is judged by the investigator.

7. Metformin contraindication according to the package instructions.

8. Currently receiving systemic corticosteroid therapy.

9. High amounts of vitamin C (greater than or equal to 1,000 mg daily), other nutraceuticals and herbs that are thought to affect glycemic control were present or taken over the last two weeks. A medical history of heart disease (including bradycardia, conduction disorder), sinoatrial node syndrome, heart failure, hypertension, heart disease) and/or physical findings of heart disease.

Each enrolled subject was allowed to receive HC plant extract of the invention (330mg, twice daily) as an additional treatment for 14.5 consecutive days, while receiving metformin routinely and without adjusting the original dosing regimen. There are no other additional drugs/treatments for type ii diabetes. Oral glucose tolerance tests were performed after administration of metformin (alone) or HC plant extract (330mg, twice daily) + metformin (14.5 days). All subjects were asked to fast for at least 10 hours prior to dosing to avoid any effects that might be caused by food intake.

Referring to fig. 8, it is shown that subjects receiving a combination treatment of metformin and the HC plant extract of the present invention significantly reduced blood glucose values (35% reduction in HC plant extract of the present invention compared to metformin alone). The results demonstrate that the HC plant extract of the present invention can act as an additional therapeutic agent for metformin.

In summary, the results of the clinical trial are consistent with the results of the animal experiments. In particular, HC plant extracts of the present invention are suitable as adjuncts to current type ii diabetes drugs (e.g., biguanides, DPP-4 inhibitors and SGLT2 inhibitors) because they synergistically lower blood glucose values when administered with metformin, sitagliptin, eggliflozin or a combination thereof.

It should be understood that the foregoing description of the embodiments is given by way of example only and that various modifications may be made by those skilled in the art. The above specification, examples and experimental results provide a complete description of the structure and use of exemplary embodiments of the invention. Although various embodiments of the present invention have been disclosed in the foregoing detailed description, it should be understood that the invention is not limited thereto, and various changes and modifications can be made by those skilled in the art without departing from the spirit and scope of the invention.

Claims

1. Use of a wild ginger flower plant extract and a blood glucose lowering agent for the preparation of a medicament for synergistically lowering the blood glucose level of an individual for treating type ii diabetes in the individual.

2. The use of claim 1, wherein the hypoglycemic agent is at least one member selected from the group consisting of dipeptidyl peptidase-4 (DPP-4) inhibitors, insulin analogs, biguanides, sulfonylureas, Thiazolidinediones (TZDs), sodium-glucose cotransporter 2(SGLT2) inhibitors, alpha-glucosidase inhibitors, and glucagon-like peptide 1(GLP-1) receptor agonists.

3. The use of claim 2, wherein the DPP-4 inhibitor is selected from the group consisting of sitagliptin, vildagliptin, saxagliptin, linagliptin, giagliptin, alagliptin, trelagliptin, alogliptin, trelagliptin, dolagliptin, augmentin, berberine and lupeol.

4. The use according to claim 3, wherein the DPP-4 inhibitor is sitagliptin.

5. The use of claim 2, wherein the biguanide is metformin, phenformin or buformin.

6. The use as claimed in claim 5, wherein the biguanide is metformin.

7. The use of claim 2, wherein the SGLT2 inhibitor is dacemaliptin, engagliflozin, canagliflozin, ivagliflozin, togagliflozin, etanergliflozin, rigagliflozin etazin etabonate, or egagliflozin.

8. The use of claim 7, wherein the SGLT2 inhibitor is eggliflozin.

9. The use according to claim 3, wherein the hypoglycemic agent is a combination of the DPP-4 inhibitor and the biguanide.

10. The use according to claim 3, wherein the hypoglycemic agent is a combination of the DPP-4 inhibitor and the SGLT2 inhibitor.

11. The use according to claim 3, wherein the hypoglycemic agent is a combination of the biguanide and the SGLT2 inhibitor.

12. The use as claimed in claim 1, wherein the wild ginger flower plant extract is prepared in a process comprising:

(a) extracting an aerial portion of a wild ginger flower with a first solvent to produce a first extract, wherein the first solvent is (1) petroleum ether, (2) n-hexane, (3) dichloromethane, (4) chloroform, (5) ethyl acetate, (6) acetone, or (7) ethanol at a concentration of 70 to 100% (v/v in water), or (8) any combination of (1) to (7);

(b) placing the first extract into a first ion exchange chromatography column;

(d) washing the first ion exchange chromatography column with an ethanol concentration of at least 70% (v/v in water) to prepare the extract of the flowers of wild ginger.

13. The use of claim 12, wherein the wild ginger flower plant extract is characterized by having substantially the High Performance Liquid Chromatography (HPLC) profile as depicted in figure 1A or figure 1B.

14. A pharmaceutical composition for treating type ii diabetes comprising:

a wild ginger flower plant extract;

a hypoglycemic agent; and

a pharmaceutically acceptable excipient.

15. The pharmaceutical composition of claim 14, wherein the hypoglycemic agent is at least one member selected from the group consisting of dipeptidyl peptidase-4 (DPP-4) inhibitors, insulin analogs, biguanides, sulfonylureas, Thiazolidinediones (TZDs), sodium-glucose cotransporter 2(SGLT2) inhibitors, alpha-glucosidase inhibitors, and glucagon-like peptide 1(GLP-1) receptor agonists.

16. The pharmaceutical composition of claim 15, wherein the DPP-4 inhibitor is selected from the group consisting of sitagliptin, vildagliptin, saxagliptin, linagliptin, giagliptin, alagliptin, trelagliptin, alogliptin, dolegliptin, augmentin, berberine and lupeol.

17. The pharmaceutical composition of claim 16, wherein the DPP-4 inhibitor is sitagliptin.

18. The pharmaceutical composition of claim 15, wherein the biguanide is metformin, phenformin, or buformin.

19. The pharmaceutical composition of claim 18, wherein the biguanide is metformin.

20. The pharmaceutical composition of claim 15, wherein the SGLT2 inhibitor is dapagliflozin, engagliflozin, canagliflozin, ivagliflozin, togagliflozin etabonate, rigagliflozin etabonate, or egagliflozin.

21. The pharmaceutical composition of claim 20, wherein the SGLT2 inhibitor is eggliflozin.

22. The pharmaceutical composition according to claim 15, wherein the hypoglycemic agent is a combination of the DPP-4 inhibitor and the biguanide.

23. The pharmaceutical composition according to claim 15, wherein the hypoglycemic agent is a combination of the DPP-4 inhibitor and the SGLT2 inhibitor.

24. The pharmaceutical composition of claim 15, wherein the hypoglycemic agent is a combination of the biguanide and the SGLT2 inhibitor.

25. The pharmaceutical composition of claim 14, wherein the wild ginger flower plant extract is characterized by having substantially the High Performance Liquid Chromatography (HPLC) profile as depicted in figure 1A or figure 1B.

26. A kit for treating type ii diabetes comprising:

a first container comprising a wild ginger flower plant extract; and

a second container comprising a hypoglycemic agent; and

an illustration providing a user with instructions for administering the extract of the flowers of wild ginger and the hypoglycemic agent to treat type II diabetes.

27. The kit of claim 26, wherein the hypoglycemic agent is at least one member selected from the group consisting of dipeptidyl peptidase-4 (DPP-4) inhibitors, insulin analogs, biguanides, sulfonylureas, Thiazolidinediones (TZDs), sodium-glucose cotransporter 2(SGLT2) inhibitors, alpha-glucosidase inhibitors, and glucagon-like peptide 1(GLP-1) receptor agonists.

28. The kit of claim 27, wherein the DPP-4 inhibitor is selected from the group consisting of sitagliptin, vildagliptin, saxagliptin, linagliptin, giagliptin, alagliptin, trelagliptin, alogliptin, trelagliptin, dolagliptin, augmentin, berberine and lupeol.

29. The kit of claim 28, wherein the DPP-4 inhibitor is sitagliptin.

30. The kit of claim 27, wherein the biguanide is metformin, phenformin, or buformin.

31. The kit of claim 30, wherein the biguanide is metformin.

32. The kit of claim 27, wherein the SGLT2 inhibitor is dapagliflozin, engagliflozin, canagliflozin, ivagliflozin, togagliflozin, etanergliflozin, rigagliflozin etazin etabonate, or egagliflozin.

33. The kit of claim 32, wherein the SGLT2 inhibitor is eggliflozin.

34. The kit according to claim 27, wherein the hypoglycemic agent is a combination of the DPP-4 inhibitor and the biguanide.

35. The kit according to claim 27, wherein the hypoglycemic agent is a combination of the DPP-4 inhibitor and the SGLT2 inhibitor.

36. The kit of claim 27, wherein the hypoglycemic agent is a combination of the biguanide and the SGLT2 inhibitor.

37. The kit of claim 27, wherein the wild ginger flower plant extract is prepared in a process comprising:

(a) extracting an aerial portion of a wild ginger flower with a first solvent to produce a first extract, wherein the solvent is (1) petroleum ether, (2) n-hexane, (3) dichloromethane, (4) chloroform, (5) ethyl acetate, (6) acetone, or (7) ethanol at a concentration of 70 to 100% (v/v in water), or (8) any combination of (1) to (7);

(b) placing the first extract into a first ion exchange chromatography column;

38. The kit of claim 37, wherein the wild ginger flower plant extract is characterized by having substantially the High Performance Liquid Chromatography (HPLC) profile as depicted in figure 1A or figure 1B.