WO2021027583A1 - 包含柠檬苦素类化合物和磺酰脲类药物的组合产品 - Google Patents

包含柠檬苦素类化合物和磺酰脲类药物的组合产品 Download PDF

Info

Publication number
WO2021027583A1
WO2021027583A1 PCT/CN2020/105728 CN2020105728W WO2021027583A1 WO 2021027583 A1 WO2021027583 A1 WO 2021027583A1 CN 2020105728 W CN2020105728 W CN 2020105728W WO 2021027583 A1 WO2021027583 A1 WO 2021027583A1
Authority
WO
WIPO (PCT)
Prior art keywords
combination product
pharmaceutically acceptable
group
limonin
acid
Prior art date
Application number
PCT/CN2020/105728
Other languages
English (en)
French (fr)
Inventor
李栋
韩铨
胡柳
薛莲
Original Assignee
浙江养生堂天然药物研究所有限公司
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 浙江养生堂天然药物研究所有限公司 filed Critical 浙江养生堂天然药物研究所有限公司
Priority to JP2022508933A priority Critical patent/JP7383126B2/ja
Priority to EP20853232.5A priority patent/EP4014999B1/en
Priority to US17/633,953 priority patent/US20220296615A1/en
Publication of WO2021027583A1 publication Critical patent/WO2021027583A1/zh

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/64—Sulfonylureas, e.g. glibenclamide, tolbutamide, chlorpropamide
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/365—Lactones
    • A61K31/366—Lactones having six-membered rings, e.g. delta-lactones
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
    • A61K31/58—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids containing heterocyclic rings, e.g. danazol, stanozolol, pancuronium or digitogenin
    • A61K31/585—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids containing heterocyclic rings, e.g. danazol, stanozolol, pancuronium or digitogenin containing lactone rings, e.g. oxandrolone, bufalin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/20—Pills, tablets, discs, rods
    • A61K9/2004—Excipients; Inactive ingredients
    • A61K9/2022—Organic macromolecular compounds
    • A61K9/205—Polysaccharides, e.g. alginate, gums; Cyclodextrin
    • A61K9/2054—Cellulose; Cellulose derivatives, e.g. hydroxypropyl methylcellulose
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics

Definitions

  • the present invention belongs to the technical field of medicine, and specifically relates to a kind of compound containing limonin (and its pharmaceutically acceptable derivatives, esters, stereoisomers, salts or prodrugs) and sulfonylurea drugs (or its pharmaceutical Acceptable derivatives).
  • the invention also relates to the use of the combination product in the treatment and/or prevention of diabetes and metabolic syndrome related diseases.
  • diabetes there were approximately 425 million people with diabetes worldwide in 2017, and 1 out of 11 people has diabetes.
  • the number of diabetes patients in China is about 110 million, ranking first in the world. It is predicted that by 2040, there will be 642 million people suffering from diabetes in the world and 151 million people with diabetes in China. Diabetes requires lifelong monitoring and treatment. If it is not well controlled, it will lead to secondary cardiovascular disease, blindness, stroke, diabetic nephropathy, diabetic gangrene and other complications, which will seriously endanger human health and life.
  • oral hypoglycemic drugs are the main treatment.
  • oral hypoglycemic drugs mainly include: sulfonylureas, biguanides, ⁇ -glycosidase inhibitors, thiazolidinediones, DPP-4 inhibitor drugs, etc., but the oral hypoglycemic drugs are prone to drug resistance Serious side effects such as sex, hypoglycemia and liver and kidney toxicity.
  • Sulfonylureas are oral hypoglycemic drugs with the earliest application, the most variety, and the most extensive clinical application. SU drugs are divided into the first and second generations.
  • the glimepiride developed in recent years It is called the third-generation SU drug for its small dosage, a certain effect of improving insulin resistance, and reducing insulin dosage, such as: glibenclamide, gliclazide, glipizide, gliquidone and Glimepiride and others.
  • the mechanism of action is: (1) Effect on pancreatic islet ⁇ cells: It is known that when SU exerts its effect on pancreatic ⁇ cells, it must first bind to the SU receptor on the surface of the ⁇ cell, and then with the ATP-sensitive potassium on the surface of the ⁇ cell. The channel is coupled to close the channel and the cell membrane is depolarized, thereby releasing insulin. Therefore, SU can stimulate ⁇ cells to release insulin, thereby reducing blood sugar. (2) Effect on the outside of the pancreas: SU can promote liver glycogen synthesis, reduce glycogen production, and slow down the release rate of liver glucose into the blood.
  • SU can increase the uptake and utilization of glucose by surrounding tissues, and increase the number of insulin receptors on the cell membrane, thereby increasing the body's insulin sensitivity.
  • the main side effects of sulfonylurea drugs are: high-dose use will have the risk of hypoglycemia (especially in elderly patients and those with liver and kidney insufficiency), blood system reactions (leukocytes, neutrophils, platelets or pancytopenia, Hemolytic anemia, etc.), allergic reactions (pruritus, urticaria, erythema, dermatitis, etc.), nervous system reactions (headache, dizziness, paresthesia, drowsiness, tinnitus, vision loss, tremor, ataxia, etc.).
  • Limonoid compounds mainly exist in the fruits of Rutaceae plants, such as Citrus aurantium, navel orange, citrus, orange, grapefruit and so on. The content in the pit (seed) is higher, and the content in the peel is less (about 1/10000 to 5/100,000). There are about 50 kinds of limonoid compounds isolated and identified from citrus plants. Limonin compounds have a variety of biological activities such as anti-tumor, insect antifeedant, anti-virus, analgesic, anti-inflammatory, and hypnotic. It can be used in functional food additives, anti-cancer foods, pesticides, feed additives, etc.
  • the present invention provides a compound comprising limonin (or its pharmaceutically acceptable derivative, its ester, its stereoisomer, its salt or prodrug) and a sulfonylurea drug (or its pharmaceutically acceptable Derivatives of ), and the composition is used to prevent or/and treat diabetes and metabolic syndrome related diseases.
  • limonin compounds mentioned in the present invention or their pharmaceutically acceptable derivatives, their esters, their stereoisomers, their salts or prodrugs
  • sulfonylurea drugs (or their pharmaceutically acceptable derivatives) Accepted derivatives compared with the same dose of sulfonylurea drugs (or pharmaceutically acceptable derivatives thereof) or limonin compounds (or pharmaceutically acceptable derivatives thereof) as monotherapy .
  • Its esters, its stereoisomers, its salts or prodrugs can significantly improve the therapeutic effect of hypoglycemic, has a synergistic effect.
  • the dosage of sulfonylurea drugs (or their pharmaceutically acceptable derivatives) is reduced, and their potential side effects are reduced.
  • a compound comprising limonin (or a pharmaceutically acceptable derivative, ester, stereoisomer, salt or prodrug thereof), and sulfonylurea
  • limonin or a pharmaceutically acceptable derivative, ester, stereoisomer, salt or prodrug thereof
  • sulfonylurea A combination product of a drug (or a pharmaceutically acceptable derivative thereof), or a combination product consisting solely of a limonin compound (or a pharmaceutically acceptable derivative, ester, stereoisomer, salt or pro- Drugs), and sulfonylurea drugs (or pharmaceutically acceptable derivatives thereof) as a combination product composed of active ingredients.
  • Limonoids mentioned in the present invention are a class of highly oxidized compounds with a 4,4,8-trimethyl-17-furan steroid skeleton or a general term of their derivatives (or can be expressed as It is composed of a variant of the polycyclic core structure of furan lactone, a compound with four fused six-membered rings and one furan ring).
  • examples of limonin compounds include but are not limited to: limonin, isolimonic acid, 7 ⁇ -limonol, obacunone, ichangin ), Yichang root Xin (ichangensin), Nomilin (nomilin), Deacetylnomilin (deacetylnomilin), Nomilin acid (nomilin acid), Deacetrynomilin acid (deacetrylnomilin acid), Citrate ( citrusin), isoobacunoic acid, etc. and any glycoside derivatives.
  • An exemplary limonin compound, that is, the structural formula of limonin is shown below.
  • glycoside derivatives of limonin compounds mentioned in the present invention include but are not limited to: limonin 17- ⁇ -glucopyranoiside, ichangin 17- ⁇ -D-glucopyranoiside ), isolimonic acid 17- ⁇ -D-glucopyranoside, deactylnomilin 17- ⁇ -D-glucopyranosied, nomilin 17- ⁇ -D-glucopyranoside ), obacunone 17- ⁇ -D-glucopynoside, nomilinic acid 17- ⁇ -D-glucopyranosid, deacetylnomilinic acid 17- ⁇ -D-glucopyranosid )Wait.
  • the limonoid compounds mentioned in the present invention exist in the form of monomers or extracts.
  • the monomers are obtained by extraction or artificial synthesis, and their sources are commercially available, or they can be easily prepared and obtained by the prior art in the field.
  • the sulfonylurea drugs mentioned in the present invention include but are not limited to glibenclamide, gliclazide, glipizide, gliquidone, glimepiride and the like.
  • the sulfonylurea drugs can exist in the form of the original compound or in the form of a pharmaceutically acceptable derivative thereof.
  • the combination product is in the form of a pharmaceutical composition
  • the pharmaceutical composition is in the form of a unit dosage.
  • the limonin compounds (or pharmaceutically acceptable derivatives, esters, stereoisomers, salts, or prodrugs thereof), and sulfonylurea drugs (or pharmaceutically acceptable derivatives thereof)
  • the above acceptable derivatives, esters, stereoisomers, salts or prodrugs thereof) are each in the form of separate preparations.
  • the limonin compound or its pharmaceutically acceptable derivative, its ester, its stereoisomer, its salt or prodrug
  • the sulfonylurea drug or its pharmaceutically acceptable The derivatives, esters, stereoisomers, salts or prodrugs thereof) can be administered simultaneously or sequentially.
  • the amount of the sulfonylurea drug is 10 mg, 20 mg, 40 mg, 50 mg, 75 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 375 mg, 500 mg , 750mg, 1500mg, 1875mg or 2000mg, and the range between each amount, the range includes but not limited to: 10mg-20mg, 10mg-40mg, 10mg-50mg, 10mg-75mg, 10mg-100mg, 10mg-150mg , 10mg-200mg, 10mg-250mg, 10mg-300mg, 10mg-375mg, 10mg-500mg, 10mg-750mg, 10mg-1500mg, 10mg-1875mg, 10mg-2000mg, 20mg-40mg, 20mg-50mg, 20mg-75mg, 20mg -100mg, 20mg-150
  • the limonin compound or its pharmaceutically acceptable derivative, its ester, its stereoisomer, its salt or prodrug) is 10mg, 20mg, 40mg, 50mg, 75mg, 100mg , 150mg, 200mg, 250mg, 300mg, 375mg, 500mg, 750mg, 1500mg, 1875mg or 2000mg, and the range of the amount between each dose, the range includes but not limited to: 10mg-20mg, 10mg-40mg, 10mg -50mg, 10mg-75mg, 10mg-100mg, 10mg-150mg, 10mg-200mg, 10mg-250mg, 10mg-300mg, 10mg-375mg, 10mg-500mg, 10mg-750mg, 10mg-1500mg, 10mg-1875mg, 10mg-2000mg , 20mg-40mg, 20mg-50mg,
  • the sulfonylurea drug is selected from glibenclamide, gliclazide, glipizide, gliquidone, glimepiride, etc.
  • the limonin compound is selected from From one or more of the following: limonin, isocitrolactone acid, 7 ⁇ -limonol, corkone, Yichang aurantine, Yichang rootsine, nomilin, deacetyl nomilin, nomilin Acid, deacetylnomilinic acid, citrin, isoflavone acid and any glycoside derivatives thereof.
  • the combination product further comprises a pharmaceutically acceptable carrier, diluent or excipient.
  • the combination product is in the form of tablets, capsules, granules, syrups, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, gas
  • the combination product in the preparation of a medicine for the prevention and/or treatment of diabetes and metabolic syndrome related diseases.
  • the diabetes is type I diabetes.
  • the diabetes is type II diabetes.
  • the limonin compounds (or pharmaceutically acceptable derivatives, esters, stereoisomers, salts or prodrugs thereof), and sulfonylureas are provided A method of combined administration of drugs (or pharmaceutically acceptable derivatives thereof) to prevent and/or treat diseases.
  • the limonin compounds or pharmaceutically acceptable derivatives, esters, stereoisomers, salts or prodrugs thereof
  • sulfonylurea drugs Or a pharmaceutically acceptable derivative thereof
  • the limonin compounds or pharmaceutically acceptable derivatives, esters, stereoisomers, salts or prodrugs thereof
  • sulfonylurea drugs Or a pharmaceutically acceptable derivative thereof
  • the limonin compounds or pharmaceutically acceptable derivatives, esters, stereoisomers, salts or prodrugs thereof
  • sulfonylurea drugs Or a pharmaceutically acceptable derivative thereof
  • the limonin compounds or pharmaceutically acceptable derivatives, esters, stereoisomers, salts or prodrugs thereof
  • sulfonylurea drugs Or a method for co-administering a pharmaceutically acceptable derivative thereof to improve leptin sensitivity.
  • the limonin compounds (or pharmaceutically acceptable derivatives, esters, stereoisomers, salts, or prodrugs thereof), and sulfonylurea drugs (or pharmaceutically acceptable derivatives thereof) can be mixed into one formulation to be administered in the form of a pharmaceutical composition (preferably, in the form of a dosage unit); in some embodiments, the limonin compound (or a pharmaceutically acceptable The derivatives, esters, stereoisomers, salts or prodrugs thereof, and sulfonylurea drugs (or pharmaceutically acceptable derivatives thereof) are each in a separate preparation form (preferably, each is in a separate In some embodiments, the limonin compound (or its pharmaceutically acceptable derivative, ester, stereoisomer, salt or prodrug thereof), and Sulfonylurea drugs (or pharmaceutically acceptable derivatives thereof are administered simultaneously; in some embodiments, the limonin compounds (or pharmaceutically acceptable derivatives, esters, and stereoisomers thereof) Body,
  • the pharmaceutical composition may be presented as needed.
  • Form preferably, dosage unit form
  • the combination product of the present invention of the drug (or its pharmaceutically acceptable derivative, its ester, its stereoisomer, its salt or prodrug), daily administration includes but not limited to: 1 time, 2 times, 3 times , 4 times, 5 times or 6 times.
  • the limonin compounds may be each in the form of a separate preparation (preferably, each in the form of a separate dosage unit) as required.
  • Daily administration includes but is not limited to: 1, 2, 3, 4, 5 or 6 times.
  • the limonin compounds (or pharmaceutically acceptable derivatives, esters, stereoisomers, salts, or prodrugs thereof), and sulfonylurea drugs (or pharmaceutically acceptable derivatives thereof) The daily dosage of the acceptable derivative is as follows: calculated according to the weight of a 60kg adult, the daily dosage of the sulfonylurea drug (or a pharmaceutically acceptable derivative thereof) is 10mg, 20mg, 40mg , 50mg, 75mg, 100mg, 150mg, 200mg, 250mg, 300mg, 375mg, 500mg, 750mg, 1500mg, 1875mg or 2000mg, and the range between the respective amounts, the range includes but not limited to: 10mg-20mg, 10mg -40mg, 10mg-50mg, 10mg-75mg, 10mg-100mg, 10mg-150mg, 10mg-200mg, 10mg-250mg, 10mg
  • the daily dosage of the limonin compound is 10mg, 20mg, 40mg, 50mg, 75mg, 100mg, 150mg, 200mg, 250mg, 300mg, 375mg, 500mg, 750mg, 1500mg, 1875mg or 2000mg, and the range of the amount between each dose, the range includes but not limited to: 10mg- 20mg, 10mg-40mg, 10mg-50mg, 10mg-75mg, 10mg-100mg, 10mg-150mg, 10mg-200mg, 10mg-250mg, 10mg-300mg, 10mg-375mg, 10mg-500mg, 10mg-750mg, 10mg-1500mg, 10mg-1875mg, 10mg-2000mg, 20mg
  • a method of preparing a combination product in the form of a pharmaceutical composition is provided.
  • the limonoid compound or its pharmaceutically acceptable derivative, its ester, its stereoisomer, and its salt it is preferable to combine the limonoid compound or its pharmaceutically acceptable derivative, its ester, its stereoisomer, and its salt according to a common method.
  • prodrugs and sulfonylurea drugs or their pharmaceutically acceptable derivatives are combined with pharmaceutically acceptable carriers, excipients, diluents and other pharmaceutical adjuvants to achieve formulation to obtain the form.
  • a kit which contains the combination product described herein.
  • pharmaceutically acceptable salt refers to a salt of a free acid or free base, usually by reacting the free base with a suitable organic or inorganic acid or by reacting the acid with a suitable organic or inorganic base. Prepare by reaction.
  • the term can be used for any compound, including limonin compounds (having the function of free acid or free base) and the like.
  • Representative salts include: acetate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, calcium edetate, camphorsulfonate, Carbonate, chloride, clavulanate, citrate, dihydrochloride, edetate, ethanedisulfonate, propionate lauric sulfate (estolate), ethanesulfonate (esylate), Fumarate, gluconate, gluconate, glutamate, glycol lylarsanilate, hexylresorcinate, hydrabamine, hydrobromide Acid salt, hydrochloride, hydroxynaphthate, iodide, isethionate, lactate, lactobionate, laurate, malate, maleate, mandelate, methanesulfonic acid Salt, methyl bromide, methyl nitrate, methosulf
  • acidic substituent such as -COOH, ammonium salt, morpholinium salt, sodium salt, potassium salt, barium salt, calcium salt, etc.
  • acid salts can be formed, such as hydrochloride, hydrobromide, phosphate, sulfate, trifluoroacetate, trichloroethyl Acid salt, acetate, oxalate, maleate, pyruvate, malonate, succinate, citrate, tartrate, fumarate, mandelate, benzoate, Cinnamate, methanesulfonate, ethanesulfonate, picrate, etc.
  • the sources of sulfonylurea drugs referred to in the present invention may include, but are not limited to: glibenclamide single tablets/capsules, gliclazide single tablets/capsules, glipizide single tablets/capsules, and Liquiquidone single tablet/capsule, glimepiride single tablet/capsule, glibenclamide metformin tablet/capsule, gliclazide metformin tablet/capsule, glipizide metformin tablet/capsule, gliquidone Metformin tablets/capsules, glimepiride metformin tablets/capsules, pioglitazone hydrochloride glimepiride tablets, glimepiride rosiglitazone maleate tablets, etc.
  • the following diabetic mouse models are selected (the models are well-known to those skilled in the art or can be easily obtained according to conventional textbooks, technical manuals and scientific literature in the field) to simulate different stages of diabetes in the human body Pathological conditions.
  • the limonin compounds mentioned in this embodiment exist in the form of monomers or extracts.
  • the monomers are obtained by extraction or artificial synthesis, and their sources are commercially available, or they can be easily prepared and obtained by the existing technology in the field.
  • ICR mice were modeled with streptozotocin (STZ) to obtain a mouse pancreatic ⁇ -cell injury model (Li Nan et al. Protective effect of pine pollen on kidney injury in mice with diabetic nephropathy, Science and Technology Review, 2014, 32 (4/5): 95-99), complete the animal hypoglycemic effect evaluation experiment (this model can simulate the state of pancreatic islet ⁇ cell damage in type I and type II diabetes).
  • STZ streptozotocin
  • Limonin compounds are selected from limonin, isocitrolactone acid, limonin glycosides, isocitrolactone acid glycosides, and a separate glibenclamide administration group, limonin administration group, and isocitrin The acid administration group, the limonin glycoside administration group, the isocitrolactone acid glycoside administration group, and the combined administration group compounded with glibenclamide.
  • mice ICR mice (20 ⁇ 2g), 6 weeks old, purchased from Zhejiang Academy of Medical Sciences, were preliminarily reared for 7 days and then subjected to experimental rearing. It should be noted that the breeding conditions for the mice are temperature 23 ⁇ 1°C, humidity 55 ⁇ 10%, light on between 7 am and 7 pm (light off at other times), and allow the mice to freely take in water and feed.
  • the experimental feed uses stable growth feed for mice (national standard M2118), and the animal security department is responsible for the daily feeding and management of animals, providing enough blocks and fresh drinking water for animals every day.
  • mice 15 male mice were randomly selected as normal control group. After 12 hours of fasting, the remaining mice were given a single intraperitoneal injection of STZ at a dose of 150 mg/kg. After 72 hours, mice with blood glucose values of 15-25 mmol/L were no difference. Grouped into the experiment, each group of 15 rats, after 2 weeks of administration, blood was taken to test the indicators.
  • Gavage dose daily gavage of limonin group 0.02g/kg, daily gavage of isocitrolactone group 0.02g/kg, daily gavage of limonin glycosides 0.02g/kg, isolimon
  • the lactone acid glycoside was given 0.02g/kg daily by gavage
  • the glibenclamide group was given 0.02g/kg glyburide daily
  • the limonin and glibenclamide group was given lemon at the same time daily.
  • Picrine 0.01g/kg and glibenclamide 0.01g/kg the combined group of isocitrolactone and glibenclamide was given daily simultaneous intragastric administration of 0.01g/kg of isocitrate and glibenclamide 0.01g /kg
  • the limonin glycoside glibenclamide combination group was given daily simultaneous oral administration of limonin glycoside 0.01g/kg and glibenclamide 0.01g/kg, combined with isocitrolactone glycoside glibenclamide
  • the group was given isocitrolactone acid glycoside 0.01g/kg and glibenclamide 0.01g/kg at the same time every day, and the gastric dose was 10mL/kg.
  • the normal group and the model group were given 10mL/kg distilled water. Two weeks later, 1 hour after the last administration, the blood glucose level was measured by cutting the tail (Johnson blood glucose meter), and calculating the average value of each group.
  • the SPSS16.0 software was used for statistical analysis, and the data were expressed as mean and standard deviation. The before and after data were analyzed by t test, and p ⁇ 0.05 was considered statistically significant. The test results are shown in Table 1 below.
  • Table 1 shows the blood glucose level of STZ mice after daily gavage for two weeks
  • limonin and its derivatives can significantly reduce the blood glucose level of STZ islet cell injury mice.
  • the combined use of limonin and its derivatives and glibenclamide has a significantly improved effect compared to the single medication, showing a synergistic effect.
  • limonin and its derivatives when used in combination with glibenclamide, they can effectively reduce the dosage of the two drugs, improve the safety of the treatment plan, and reduce side effects when the hypoglycemic effect is equivalent. .
  • db/db mice (strain name BKS.Cg-Dock7 m+/+ Lepr db /Nju) were used to complete the animal hypoglycemic effect evaluation experiment (blood sugar level and leptin).
  • Limonin compounds choose corkone, isoflavone acid and corkone glycosides, and set up separate gliclazide administration group, corkone administration group, isoflavone acid administration group, and corkone glycoside administration group respectively And the combined administration group with Gliclazide.
  • mice As a type II diabetes model mouse, 6-week-old SPF db/db mice were purchased from Nanjing Model Biology Institute, and the experimental breeding was carried out after 7 days of preliminary breeding. It should be noted that the breeding conditions for the mice are temperature 23 ⁇ 1°C, humidity 55 ⁇ 10%, light on between 7 am and 7 pm (light off at other times), and allow the mice to freely take in water and feed.
  • the experimental feed uses stable growth feed for mice (national standard M2118), and the animal security department is responsible for the daily feeding and management of animals, providing enough blocks and fresh drinking water for animals every day.
  • mice select male db/db mice (20 ⁇ 2g), each group of 18 male mice for experiment.
  • Gavage dose the corkone group was given cork ketone 0.04 g/kg daily, the isoflavone acid group was given isoflavone acid 0.04 g/kg every day, the corkone glycoside group was given corkone glycosides daily Gliclazide 0.04g/kg and Gliclazide group were given Gliclazide 0.04g/kg per day, and Gliclazide combination group was given Gliclazide 0.02g/kg and Gliclazide.
  • Gliclazide isoflavone combination group was given 0.02g/kg isoflavone acid and 0.02g/kg Gliclazide by gavage at the same time every day, Gliclazide combination group of corvone glycoside
  • Table 2 shows the blood glucose level and leptin of db/db mice after daily intragastric administration for two weeks
  • corkstone and its derivatives can significantly reduce the blood glucose level of db/db diabetic mice.
  • the combined use of phellodendone and its derivatives and gliclazide have significantly improved effects compared with single use, showing a synergistic effect.
  • corvone and its derivatives are used in combination with gliclazide, compared to the single administration, when the hypoglycemic effect is equivalent, the dosage of both can be effectively reduced, the safety of the treatment plan can be improved, and the side effects can be reduced.
  • limonin compounds represented by corvone and its derivatives can significantly improve leptin sensitivity, especially when used in combination with gliclazide, can significantly improve the utilization efficiency of leptin in the body and improve the body's glucose metabolism function , Improve the related functions of glucose metabolism in diabetic mice.
  • ICR mice were modeled with streptozotocin (STZ) to obtain a mouse pancreatic ⁇ -cell injury model (Li Nan et al. Protective effect of pine pollen on kidney injury in mice with diabetic nephropathy, Science and Technology Review, 2014, 32 (4/5): 95-99), complete the animal hypoglycemic effect evaluation experiment (this model can simulate the state of pancreatic islet ⁇ cell damage in type I and type II diabetes).
  • STZ streptozotocin
  • Limonin compounds were selected from Yichang aurantine, Yichang aurantine, and Yichang aurantine glycosides, and a separate glipizide administration group, Yichang aurantine administration group, Yichang aurantine administration group, and Yichang aurantine administration group were set up.
  • the drug glycoside administration group and the combined administration group combined with glipizide.
  • mice ICR mice (20 ⁇ 2g), 6 weeks old, purchased from Zhejiang Academy of Medical Sciences, were preliminarily reared for 7 days and then subjected to experimental rearing. It should be noted that the breeding conditions for the mice are temperature 23 ⁇ 1°C, humidity 55 ⁇ 10%, light on between 7 am and 7 pm (light off at other times), and allow the mice to freely take in water and feed.
  • the experimental feed uses stable growth feed for mice (national standard M2118), and the animal security department is responsible for the daily feeding and management of animals, providing enough blocks and fresh drinking water for animals every day.
  • mice 15 male mice were randomly selected as normal control group. After 12 hours of fasting, the remaining mice were given a single intraperitoneal injection of STZ at a dose of 150 mg/kg. After 72 hours, mice with blood glucose values of 15-25 mmol/L were no difference. Grouped into the experiment, each group of 15 rats, after 2 weeks of administration, blood was taken to test the indicators.
  • Gavage dose Yichang aurantine group was given 0.1g/kg daily, Yichang Genxin group was given 0.1g/kg daily, and Yichang aurantine glycosides were given daily 0.1g/kg, glied
  • the pizazine group was given glipizide 0.1g/kg daily, and the Yichang serotonin and glipizide combined group were given Yichang serotonin 0.05g/kg and glipizide 0.05g/day simultaneously.
  • Yichang root sinoglycoside and glipizide combined group were given Yichang roots 0.05g/kg and glipizide 0.05g/kg at the same time every day, and Yichang sinoglycoside glipizide combination group was given daily at the same time Yichang aurin glucoside 0.05g/kg and glipizide 0.05g/kg were administered intragastrically, and the intragastric dose was 10mL/kg.
  • the normal group and the model group were given 10mL/kg distilled water. Two weeks later, 1 hour after the last administration, the blood glucose level was measured by cutting the tail (Johnson blood glucose meter), and calculating the average value of each group.
  • the SPSS16.0 software was used for statistical analysis, and the data were expressed as mean and standard deviation. The before and after data were analyzed by t test, and p ⁇ 0.05 was considered statistically significant. The test results are shown in Table 3 below.
  • Table 3 shows the blood glucose level of STZ mice after daily gavage for two weeks
  • the three limonin compounds can significantly reduce the blood glucose level of STZ islet cell injury model mice.
  • the effect is significantly improved compared to the effect of single medication, which is similar to the blood glucose level of normal mice, showing a synergistic effect.
  • the above-mentioned three limonin compounds when used in combination with glipizide, they can effectively reduce the dosage of the two drugs, improve the safety of the treatment plan, and reduce side effect.
  • nomilin, deacetyl nomilin, nomilin acid, deacetyl nomilin acid glycosides are selected for the limonin compounds, and separate gliquidone administration groups and nomilin are set respectively.
  • mice As a type II diabetes model mouse, 6-week-old SPF db/db mice were purchased from Nanjing Model Biology Institute, and the experimental breeding was carried out after 7 days of preliminary breeding. It should be noted that the breeding conditions for the mice are temperature 23 ⁇ 1°C, humidity 55 ⁇ 10%, light on between 7 am and 7 pm (light off at other times), and allow the mice to freely take in water and feed.
  • the experimental feed uses stable growth feed for mice (national standard M2118), and the animal security department is responsible for the daily feeding and management of animals, providing enough blocks and fresh drinking water for animals every day.
  • mice male db/db mice (20 ⁇ 2g) were selected, 18 male mice in each group were tested, and the drinking bottle was sterilized once a week.
  • Gavage dose Nomilin group was given nomilin 0.2g/kg daily, deacetyl nomilin group was given deacetyl nomilin 0.2g/kg daily, nomilin acid group was given daily
  • the stomach was given nomilin acid 0.2g/kg, the deacetylnomilin acid glycoside group was given daily intragastric administration of deacetylnomilin acid glycoside 0.2g/kg, and the gliquidone group was given daily gliquidone
  • the 0.2g/kg, nomilin combination group was given nomilin 0.1g/kg and gliquidone 0.1g/kg at the same time every day, and the nomilin acid combination group was given nomilin every day at the same time
  • Acid 0.1g/kg and gliquidone 0.1g/kg, the deacetyl-nomilin combination group was given deacetyl-nomilin 0.1g/kg and gliquidone 0.1g/kg at the same time every day, deacetylation
  • Table 4 shows the blood glucose level and insulin of db/db mice after two weeks of intragastric administration every day
  • nomilin and its derivatives can significantly reduce the blood sugar level of db/db diabetic mice, whether administered alone or in combination with gliquidone.
  • the combined use of nomilin and its derivatives and gliquidone has a significantly improved effect compared to the single medication, showing a synergistic effect.
  • nomilin and its derivatives when used in combination with gliquidone, they can effectively reduce the dosage of the two drugs, improve the safety of the treatment plan, and reduce the side effects when the hypoglycemic effect is equivalent. .
  • limonin compounds represented by nomilin and its derivatives can significantly improve insulin sensitivity, especially when used in combination with gliquidone, it can significantly improve the utilization efficiency of insulin in the body and improve the body's glucose metabolism. Improve the related functions of glucose metabolism in diabetic mice.
  • Limonin compounds choose nomilin glycosides, deacetyl nomilin glycosides, and nomilin acid glycosides, and set up a separate glimepiride administration group, nomilin glycoside administration group, and deacetyl nomilin glycoside administration The drug group, the nomilin acid glycoside administration group and the combined administration group combined with glimepiride.
  • mice ICR mice (20 ⁇ 2g), 6 weeks old, purchased from Zhejiang Academy of Medical Sciences, were preliminarily reared for 7 days and then subjected to experimental rearing. It should be noted that the breeding conditions for the mice are temperature 23 ⁇ 1°C, humidity 55 ⁇ 10%, light on between 7 am and 7 pm (light off at other times), and allow the mice to freely take in water and feed.
  • the experimental feed uses stable growth feed for mice (national standard M2118), and the animal security department is responsible for the daily feeding and management of animals, providing enough blocks and fresh drinking water for animals every day.
  • mice Take male mice, 15 randomly as the normal control group, and other mice have a high-fat diet (high-fat diet formula: cholesterol 1%, egg yolk powder 10%, lard 10%, basic diet 79% to establish obesity Mouse model) STZ was injected intraperitoneally at 35mg/kg for three consecutive days for four consecutive days. After one week, fasting blood glucose was measured and fasting blood glucose was measured. The mice with blood glucose values of 15-25mmol/L were selected and included in the experiment without difference. Fat diet, 15 rats in each group, after 2 weeks of administration, blood was taken to test the indicators.
  • high-fat diet formula: cholesterol 1%, egg yolk powder 10%, lard 10%, basic diet 79% to establish obesity Mouse model
  • Gavage dose daily intragastric administration of 0.5g/kg for nomilin glycoside group, daily intragastric administration of 0.5g/kg for deacetyl nomilin glycoside group and 0.5g/kg daily for nomilin acid glycosides , Glimepiride group was given Glimepiride 0.5g/kg daily by gavage, and Glimepiride combination group was given Glimepiride 0.25g/kg and Glimepiride at the same time daily.
  • deacetyl nomilin glycoside and glimepiride combination group was given deacetyl nomilin glycoside 0.25g/kg and glimepiride 0.25g/kg at the same time every day, nomilin acid glycoside grid
  • the Limepiride combination group was given nomilin acid glycoside 0.25g/kg and glimepiride 0.25g/kg at the same time every day, and the gastric volume was 10mL/kg.
  • the normal group and the model group were given 10mL/kg distilled water.
  • the SPSS16.0 software was used for statistical analysis, and the data were expressed as mean and standard deviation. The before and after data were analyzed by t test, and p ⁇ 0.05 was considered statistically significant. The test results are shown in Table 5 below.
  • Table 5 shows the blood glucose level of STZ mice after daily gavage for two weeks
  • the above three limonoid glycosides can significantly reduce the blood glucose level of STZ type II diabetes model mice.
  • the effect is significantly improved compared to the single drug, and the blood sugar level is similar to that of normal mice, showing a synergistic effect.
  • the above three limonin glycosides when used in combination with glimepiride, they can effectively reduce the dosage of the two drugs, improve the safety of the treatment plan, and reduce side effect.
  • a method for preparing tablets of the combination product (nomilin and glibenclamide) of the present invention is exemplarily provided.
  • a single tablet contains the following ingredients: 50mg Nomilin, 400mg Glyburide Hydrochloride, 20mg Hydroxypropyl Methyl Cellulose, 30mg Sodium Carboxymethyl Cellulose, and Microcrystalline Cellulose 20mg, magnesium stearate 5.2mg, Opadry 20.8mg, a total of 1000 tablets.
  • the preparation method includes the following steps:

Abstract

一种含有柠檬苦素类化合物（以及其药学上可接受的衍生物、酯、立体异构体、盐或前药）和磺酰脲类药物（格列本脲、格列齐特、格列呲嗪、格列喹酮和格列美脲等）的组合产品。该组合产品用于治疗和/或预防与糖尿病相关疾病等。

Description

包含柠檬苦素类化合物和磺酰脲类药物的组合产品 技术领域
本发明属于医药技术领域，具体涉及一种含有柠檬苦素类化合物(及其药学上可接受的衍生物、酯、立体异构体、盐或前药)和磺酰脲类药物(或其药学上可接受的衍生物)的组合产品。本发明还涉及该组合产品在治疗和/或预防糖尿病以及代谢综合症相关疾病的用途。
背景技术
根据IDF统计，2017年全球糖尿病患者约有4.25亿人，每11个人就有1人患有糖尿病。中国糖尿患者人数约1.1亿人，位居世界第一。预测到2040年，全球将会有6.42亿人患有糖尿病，中国糖尿病患者也将达到1.51亿。糖尿病需终身监测及治疗，如得不到良好控制，将会导致患者继发心血管疾病，失明、脑卒中、糖尿病肾病、糖尿病坏疽等并发症，严重危害人的健康和生命。
糖尿病中90％以上是Ⅱ型糖尿病，以口服降糖药为主要治疗方式。目前，口服降糖药物包括：磺酰脲类、双胍类、α-糖苷酶抑制剂、噻唑烷二酮类、DPP-4抑制剂药物等为主，但所述口服降糖药物容易出现耐药性、低血糖以及肝肾毒性等严重副作用。
磺酰脲类药物(sulfonylureas，SU)是应用最早、品种最多、临床应用也最广泛的口服降糖药，SU类药物有第一代和第二代之分，近年研制的格列美脲则以其用药剂量小、具有一定的改善胰岛素抵抗作用、减少胰岛素用量而被称为第三代SU类药物，例如：格列本脲、格列齐特、格列呲嗪、格列喹酮和格列美脲等。其作用机理为：(1)对胰岛β细胞的作用：已知SU在发挥对胰岛β细胞的作用时，必须先与β细胞表面的SU受体相结合，然后与β细胞表面的ATP敏感钾通道藕联，使此通道关闭，细胞膜去极化，从而释放胰岛素。因此，SU能刺激β细胞释放胰岛素，从而降低血糖。(2)对胰外的作用：SU可以促进肝糖原合成，减少肝糖的产生，并能减缓肝脏葡萄糖向血液中的释放速率。同时，SU可使周围组织对葡萄糖的摄取、利用增加，并可增加细胞膜上胰岛素受体的数量，从而使机体的胰岛素敏感性增加。磺酰脲类药物的副作用主要为：大剂量使用会有低血糖的风险(特别是老年患者和肝、肾功能不全者)、血液系统反应(白细胞、中性粒细胞、血小板或全 血细胞减少、溶血性贫血等)、过敏反应(皮肤瘙痒、荨麻疹、红斑、皮炎等)、神经系统反应(头痛、头晕、感觉异常、嗜睡、耳鸣、视力减退、震颤、共济失调等)。
柠檬苦素类化合物主要存在于芸香科植物果实中，如枳实、脐橙、柑桔、香橙、柚等中。以果核(种子)中含量较高，果皮中含量较少(约万分之一至十万分之五)。从柑桔属植物中分离和鉴定的柠檬苦素类化合物约50多种。柠檬苦素类化合物具有抗肿瘤、昆虫拒食、抗病毒、镇痛、抗炎、催眠等多种生物活性。可用于功能性食品添加剂、抗癌食品、杀虫剂、饲料添加剂等。
考虑到磺酰脲类药物的降糖效果和副作用，亟需找到服用方式简单、效果良好、副作用低的药物组合产品。
发明内容
本发明提供一种包含柠檬苦素类化合物(或其药学上可接受的衍生物、其酯、其立体异构体、其盐或前药)和磺酰脲类药物(或其药学上可接受的衍生物)的组合产品，以及该组合物用于预防或/和治疗糖尿病以及代谢综合症相关疾病。本发明所提及的包含柠檬苦素类化合物(或其药学上可接受的衍生物、其酯、其立体异构体、其盐或前药)和磺酰脲类药物(或其药学上可接受的衍生物)的组合产品，相较相同剂量的作为单一疗法的磺酰脲类药物(或其药学上可接受的衍生物)或柠檬苦素类化合物(或其药学上可接受的衍生物、其酯、其立体异构体、其盐或前药)，可以显著提升降糖的治疗效果，具有协同增效作用。同时，降低了磺酰脲类药物(或其药学上可接受的衍生物)的用量，降低其潜在的副作用。
在本发明的第一方面，提供了一种包含柠檬苦素类化合物(或其药学上可接受的衍生物、其酯、其立体异构体、其盐或前药)，以及磺酰脲类药物(或其药学上可接受的衍生物)的组合产品，或一种仅仅由柠檬苦素类化合物(或其药学上可接受的衍生物、其酯、其立体异构体、其盐或前药)，以及磺酰脲类药物(或其药学上可接受的衍生物)作为活性成分所组成的组合产品。
本发明所提及的柠檬苦素类化合物(Limonoids)是一类高度氧化的具有4,4,8-三甲基-17-呋喃甾体骨架的化合物或其衍生物的总称(或可表达为由呋喃内酯多环核心结构上的变体构成，具有四个稠合六元环和一个呋喃环的化合物)。具体地，柠檬苦素类化合物的实例包括但不限于：柠檬苦素(limomin)、异柠檬内酯酸(isolimonic acid)、7α-柠檬苦醇、 黄柏酮(obacunone)、宜昌橙苦素(ichangin)、宜昌根辛(ichangensin)、诺米林(nomilin)、脱乙酰诺米林(deacetylnomilin)、诺米林酸(nomilin acid)、脱乙酰诺米林酸(deacetrylnomilin acid)、枸橼苦素(citrusin)、异黄柏酮酸(isoobacunoic acid)等及其任何的糖苷衍生物。示例性柠檬苦素类化合物，即，柠檬苦素的结构式示于下文。
Figure PCTCN2020105728-appb-000001
柠檬苦素的结构式
进一步地，本发明所提及的柠檬苦素类化合物的糖苷衍生物包括但不限于：柠檬苦素糖苷(limonin 17-β-glucopyranoiside)，宜昌橙苦素糖苷(ichangin 17-β-D-glucopyranoiside)，异柠檬内酯酸糖苷(isolimonic acid 17-β-D-glucopyranoside)，脱乙酰基诺米林糖苷(deactylnomilin 17-β-D-glucopyranosied)，诺米林糖苷(nomilin 17-β-D-glucopyranoside)，黄柏酮糖苷(obacunone 17-β-D-glucopynoside)，诺米林酸糖苷(nomilinic acid 17-β-D-glucopyranosid)，脱乙酰基诺米林酸糖苷(deacetylnomilinic acid 17-β-D-glucopyranosid)等。
在一些实施方式中，本发明所提及的柠檬苦素类化合物的存在形式呈单体或提取物的形式。单体是提取得到的或人工合成的，其来源可以商购，或通过本领域现有技术它们能够容易地制备和得到的。
本发明中所提及的磺酰脲类药物包括但不限于格列本脲、格列齐特、格列呲嗪、格列喹酮和格列美脲等。磺酰脲类药物可以原化合物形式存在，或者通过其药学上可接受的衍生物的形式存在。
在一些实施方式中，所述组合产品呈药物组合物的形式，且所述药物组合物是单元剂量形式。
在一些实施方式中，所述柠檬苦素类化合物(或其药学上可接受的衍生物、其酯、其立体异构体、其盐或前药)，以及磺酰脲类药物(或其药学上可接受的衍生物、其酯、其立体异构体、其盐或前药)各自呈单独的制剂形式。进一步地，所述柠檬苦素类化合物(或 其药学上可接受的衍生物、其酯、其立体异构体、其盐或前药)，以及磺酰脲类药物(或其药学上可接受的衍生物、其酯、其立体异构体、其盐或前药)各自呈单独的单元剂量形式。进一步地，所述柠檬苦素类化合物(或其药学上可接受的衍生物、其酯、其立体异构体、其盐或前药)，以及磺酰脲类药物(或其药学上可接受的衍生物、其酯、其立体异构体、其盐或前药)可以同时或先后给药。
在一些实施方式中，所述磺酰脲类药物(或其药学上可接受的衍生物)的量是10mg、20mg、40mg、50mg、75mg、100mg、150mg、200mg、250mg、300mg、375mg、500mg、750mg、1500mg、1875mg或2000mg，以及所述各量之间的范围，所述范围包括但不限于：10mg-20mg、10mg-40mg、10mg-50mg、10mg-75mg、10mg-100mg、10mg-150mg、10mg-200mg、10mg-250mg、10mg-300mg、10mg-375mg、10mg-500mg、10mg-750mg、10mg-1500mg、10mg-1875mg、10mg-2000mg、20mg-40mg、20mg-50mg、20mg-75mg、20mg-100mg、20mg-150mg、20mg-200mg、20mg-250mg、20mg-300mg、20mg-375mg、20mg-500mg、20mg-750mg、20mg-1500mg、20mg-1875mg、20mg-2000mg、40mg-50mg、40mg-75mg、40mg-100mg、40mg-150mg、40mg-200mg、40mg-250mg、40mg-300mg、40mg-375mg、40mg-500mg、40mg-750mg、40mg-1500mg、40mg-1875mg、40mg-2000mg、50mg-75mg、50mg-100mg、50mg-150mg、50mg-200mg、50mg-250mg、50mg-300mg、50mg-375mg、50mg-500mg、50mg-750mg、50mg-1500mg、50mg-1875mg、50mg-2000mg、75mg-100mg、75mg-150mg、75mg-200mg、75mg-250mg、75mg-300mg、75mg-375mg、75mg-500mg、75mg-750mg、75mg-1500mg、75mg-1875mg、75mg-2000mg、100mg-150mg、100mg-200mg、100mg-250mg、100mg-300mg、100mg-375mg、100mg-500mg、100mg-750mg、100mg-1500mg、100mg-1875mg、100mg-2000mg、150mg-200mg、150mg-250mg、150mg-300mg、150mg-375mg、150mg-500mg、150mg-750mg、150mg-1500mg、150mg-1875mg、150mg-2000mg、200mg-250mg、200mg-300mg、200mg-375mg、200mg-500mg、200mg-750mg、200mg-1500mg、200mg-1875mg、200mg-2000mg、250mg-300mg、250mg-375mg、250mg-500mg、250mg-750mg、250mg-1500mg、250mg-1875mg、250mg-2000mg、300mg-375mg、300mg-500mg、300mg-750mg、300mg-1500mg、300mg-1875mg、300mg-2000mg、375mg-500mg、375mg-750mg、375mg-1500mg、375mg-1875mg、375mg-2000mg、500mg-750mg、500mg-1500mg、500mg-1875mg、500mg-2000mg、750mg-1500mg、750mg-1875mg、750mg-2000mg和1875mg-2000mg。。
在一些实施方式中，所述柠檬苦素类化合物或其药学上可接受的衍生物、其酯、其立体异构体、其盐或前药)是10mg、20mg、40mg、50mg、75mg、100mg、150mg、200mg、250mg、300mg、375mg、500mg、750mg、1500mg、1875mg或2000mg，以及所述各剂量之间的量的范围，所述范围包括但不限于：10mg-20mg、10mg-40mg、10mg-50mg、10mg-75mg、10mg-100mg、10mg-150mg、10mg-200mg、10mg-250mg、10mg-300mg、10mg-375mg、10mg-500mg、10mg-750mg、10mg-1500mg、10mg-1875mg、10mg-2000mg、20mg-40mg、20mg-50mg、20mg-75mg、20mg-100mg、20mg-150mg、20mg-200mg、20mg-250mg、20mg-300mg、20mg-375mg、20mg-500mg、20mg-750mg、20mg-1500mg、20mg-1875mg、20mg-2000mg、40mg-50mg、40mg-75mg、40mg-100mg、40mg-150mg、40mg-200mg、40mg-250mg、40mg-300mg、40mg-375mg、40mg-500mg、40mg-750mg、40mg-1500mg、40mg-1875mg、40mg-2000mg、50mg-75mg、50mg-100mg、50mg-150mg、50mg-200mg、50mg-250mg、50mg-300mg、50mg-375mg、50mg-500mg、50mg-750mg、50mg-1500mg、50mg-1875mg、50mg-2000mg、75mg-100mg、75mg-150mg、75mg-200mg、75mg-250mg、75mg-300mg、75mg-375mg、75mg-500mg、75mg-750mg、75mg-1500mg、75mg-1875mg、75mg-2000mg、100mg-150mg、100mg-200mg、100mg-250mg、100mg-300mg、100mg-375mg、100mg-500mg、100mg-750mg、100mg-1500mg、100mg-1875mg、100mg-2000mg、150mg-200mg、150mg-250mg、150mg-300mg、150mg-375mg、150mg-500mg、150mg-750mg、150mg-1500mg、150mg-1875mg、150mg-2000mg、200mg-250mg、200mg-300mg、200mg-375mg、200mg-500mg、200mg-750mg、200mg-1500mg、200mg-1875mg、200mg-2000mg、250mg-300mg、250mg-375mg、250mg-500mg、250mg-750mg、250mg-1500mg、250mg-1875mg、250mg-2000mg、300mg-375mg、300mg-500mg、300mg-750mg、300mg-1500mg、300mg-1875mg、300mg-2000mg、375mg-500mg、375mg-750mg、375mg-1500mg、375mg-1875mg、375mg-2000mg、500mg-750mg、500mg-1500mg、500mg-1875mg、500mg-2000mg、750mg-1500mg、750mg-1875mg、750mg-2000mg和1875mg-2000mg。
在一些实施方式中，所述磺酰脲类药物选自格列本脲、格列齐特、格列呲嗪、格列喹酮和格列美脲等，且所述柠檬苦素类化合物选自下述的一个或多个：柠檬苦素、异柠檬内酯酸、7α-柠檬苦醇、黄柏酮、宜昌橙苦素、宜昌根辛、诺米林、脱乙酰诺米林、诺米林酸、脱乙酰诺米林酸、枸橼苦素、异黄柏酮酸及其任何的糖苷衍生物。
在一些实施方式中，所述组合产品还包含药学上可接受的载体、稀释剂或赋形剂。
在一些实施方式中，所述组合产品呈片剂、胶囊剂、颗粒剂、糖浆剂、粉剂、锭剂、药囊、扁囊剂、酏剂、混悬剂、乳剂、溶液、糖浆剂、气雾剂、软膏剂、乳膏剂和注射剂的形式。
在本发明的第二方面，提供了所述组合产品在制备药物中的用途，所述药物用于预防和/或治疗糖尿病以及代谢综合症相关疾病。在一些实施方式中，所述糖尿病是I型糖尿病。在一些实施方式中，所述糖尿病是II型糖尿病。
在本发明的第三方面，提供了将所述柠檬苦素类化合物(或其药学上可接受的衍生物、其酯、其立体异构体、其盐或前药)，以及磺酰脲类药物(或其药学上可接受的衍生物)联合给药以预防和/或治疗疾病的方法。在一些实施方式中，提供了将所述柠檬苦素类化合物(或其药学上可接受的衍生物、其酯、其立体异构体、其盐或前药)，以及磺酰脲类药物(或其药学上可接受的衍生物)联合给药，以预防和/或治疗糖尿病以及代谢综合症相关疾病的方法。在一些实施方式中，提供了将所述柠檬苦素类化合物(或其药学上可接受的衍生物、其酯、其立体异构体、其盐或前药)，以及磺酰脲类药物(或其药学上可接受的衍生物)联合给药降低血糖的方法。在一些实施方式中，提供了将所述柠檬苦素类化合物(或其药学上可接受的衍生物、其酯、其立体异构体、其盐或前药)，以及磺酰脲类药物(或其药学上可接受的衍生物)联合给药改善胰岛素敏感性的方法。在一些实施方式中，提供了将所述柠檬苦素类化合物(或其药学上可接受的衍生物、其酯、其立体异构体、其盐或前药)，以及磺酰脲类药物(或其药学上可接受的衍生物联合给药以改善瘦素敏感性的方法。
在一些实施方式中，所述柠檬苦素类化合物(或其药学上可接受的衍生物、其酯、其立体异构体、其盐或前药)，以及磺酰脲类药物(或其药学上可接受的衍生物)可以混合成一个制剂中呈药物组合物的形式(优选地，剂量单元形式)给药；在一些实施方式中，所述柠檬苦素类化合物(或其药学上可接受的衍生物、其酯、其立体异构体、其盐或前药)，以及磺酰脲类药物(或其药学上可接受的衍生物)各自呈单独的制剂形式(优选地，各自呈单独的剂量单元形式)分别给药；在一些实施方式中，所述柠檬苦素类化合物(或其药学上可接受的衍生物、其酯、其立体异构体、其盐或前药)，以及磺酰脲类药物(或其药学上可接受的衍生物同时给药；在一些实施方式中，所述柠檬苦素类化合物(或其药学上可接受的衍生物、其酯、其立体异构体、其盐或前药)，以及磺酰脲类药物(或其药学上可接受的衍生物)以一个紧接着另一个的方式先后给药；在一些实施方式中，所述柠檬苦 素类化合物(或其药学上可接受的衍生物、其酯、其立体异构体、其盐或前药)，以及磺酰脲类药物(或其药学上可接受的衍生物)先后给药的时间间隔可为约30分钟、或约1小时、或约2小时、或约4小时、或约8小时、或约12小时。在一些实施方式中，可以根据需要，将所述呈药物组合物的形式(优选地，剂量单元形式)的含有所述柠檬苦素类化合物(或其药学上可接受的衍生物、其酯、其立体异构体、其盐或前药)，以及磺酰脲类药物(或其药学上可接受的衍生物、其酯、其立体异构体、其盐或前药)的本发明的组合产品，每天给药包括但不限于：1次、2次、3次、4次、5次或6次。在一些实施方式中，可以根据需要，将所述各自呈单独的制剂形式(优选地，各自呈单独的剂量单元形式)的含有所述柠檬苦素类化合物(或其药学上可接受的衍生物、其酯、其立体异构体、其盐或前药)，以及磺酰脲类药物(或其药学上可接受的衍生物)的本发明的组合产品，每天给药包括但不限于：1次、2次、3次、4次、5次或6次。
在一些实施方式中，所述柠檬苦素类化合物(或其药学上可接受的衍生物、其酯、其立体异构体、其盐或前药)，以及磺酰脲类药物(或其药学上可接受的衍生物)每日给药量如下：按照60kg成人体重计算，所述磺酰脲类药物(或其药学上可接受的衍生物)的每日给药量是10mg、20mg、40mg、50mg、75mg、100mg、150mg、200mg、250mg、300mg、375mg、500mg、750mg、1500mg、1875mg或2000mg，以及所述各量之间的范围，所述范围包括但不限于：10mg-20mg、10mg-40mg、10mg-50mg、10mg-75mg、10mg-100mg、10mg-150mg、10mg-200mg、10mg-250mg、10mg-300mg、10mg-375mg、10mg-500mg、10mg-750mg、10mg-1500mg、10mg-1875mg、10mg-2000mg、20mg-40mg、20mg-50mg、20mg-75mg、20mg-100mg、20mg-150mg、20mg-200mg、20mg-250mg、20mg-300mg、20mg-375mg、20mg-500mg、20mg-750mg、20mg-1500mg、20mg-1875mg、20mg-2000mg、40mg-50mg、40mg-75mg、40mg-100mg、40mg-150mg、40mg-200mg、40mg-250mg、40mg-300mg、40mg-375mg、40mg-500mg、40mg-750mg、40mg-1500mg、40mg-1875mg、40mg-2000mg、50mg-75mg、50mg-100mg、50mg-150mg、50mg-200mg、50mg-250mg、50mg-300mg、50mg-375mg、50mg-500mg、50mg-750mg、50mg-1500mg、50mg-1875mg、50mg-2000mg、75mg-100mg、75mg-150mg、75mg-200mg、75mg-250mg、75mg-300mg、75mg-375mg、75mg-500mg、75mg-750mg、75mg-1500mg、75mg-1875mg、75mg-2000mg、100mg-150mg、100mg-200mg、100mg-250mg、100mg-300mg、100mg-375mg、100mg-500mg、100mg-750mg、100mg-1500mg、100mg-1875mg、100mg-2000mg、150mg-200mg、150mg-250mg、150mg-300mg、150mg-375mg、150mg-500mg、150mg-750mg、 150mg-1500mg、150mg-1875mg、150mg-2000mg、200mg-250mg、200mg-300mg、200mg-375mg、200mg-500mg、200mg-750mg、200mg-1500mg、200mg-1875mg、200mg-2000mg、250mg-300mg、250mg-375mg、250mg-500mg、250mg-750mg、250mg-1500mg、250mg-1875mg、250mg-2000mg、300mg-375mg、300mg-500mg、300mg-750mg、300mg-1500mg、300mg-1875mg、300mg-2000mg、375mg-500mg、375mg-750mg、375mg-1500mg、375mg-1875mg、375mg-2000mg、500mg-750mg、500mg-1500mg、500mg-1875mg、500mg-2000mg、750mg-1500mg、750mg-1875mg、750mg-2000mg和1875mg-2000mg。按照60kg成人体重计算，所述柠檬苦素类化合物(或其药学上可接受的衍生物、其酯、其立体异构体、其盐或前药)的每日给药量是10mg、20mg、40mg、50mg、75mg、100mg、150mg、200mg、250mg、300mg、375mg、500mg、750mg、1500mg、1875mg或2000mg，以及所述各剂量之间的量的范围，所述范围包括但不限于：10mg-20mg、10mg-40mg、10mg-50mg、10mg-75mg、10mg-100mg、10mg-150mg、10mg-200mg、10mg-250mg、10mg-300mg、10mg-375mg、10mg-500mg、10mg-750mg、10mg-1500mg、10mg-1875mg、10mg-2000mg、20mg-40mg、20mg-50mg、20mg-75mg、20mg-100mg、20mg-150mg、20mg-200mg、20mg-250mg、20mg-300mg、20mg-375mg、20mg-500mg、20mg-750mg、20mg-1500mg、20mg-1875mg、20mg-2000mg、40mg-50mg、40mg-75mg、40mg-100mg、40mg-150mg、40mg-200mg、40mg-250mg、40mg-300mg、40mg-375mg、40mg-500mg、40mg-750mg、40mg-1500mg、40mg-1875mg、40mg-2000mg、50mg-75mg、50mg-100mg、50mg-150mg、50mg-200mg、50mg-250mg、50mg-300mg、50mg-375mg、50mg-500mg、50mg-750mg、50mg-1500mg、50mg-1875mg、50mg-2000mg、75mg-100mg、75mg-150mg、75mg-200mg、75mg-250mg、75mg-300mg、75mg-375mg、75mg-500mg、75mg-750mg、75mg-1500mg、75mg-1875mg、75mg-2000mg、100mg-150mg、100mg-200mg、100mg-250mg、100mg-300mg、100mg-375mg、100mg-500mg、100mg-750mg、100mg-1500mg、100mg-1875mg、100mg-2000mg、150mg-200mg、150mg-250mg、150mg-300mg、150mg-375mg、150mg-500mg、150mg-750mg、150mg-1500mg、150mg-1875mg、150mg-2000mg、200mg-250mg、200mg-300mg、200mg-375mg、200mg-500mg、200mg-750mg、200mg-1500mg、200mg-1875mg、200mg-2000mg、250mg-300mg、250mg-375mg、250mg-500mg、250mg-750mg、250mg-1500mg、250mg-1875mg、250mg-2000mg、300mg-375mg、300mg-500mg、300mg-750mg、300mg-1500mg、300mg-1875mg、300mg-2000mg、375mg-500mg、375mg-750mg、375mg-1500mg、375mg-1875mg、375mg-2000mg、500mg-750mg、500mg-1500mg、500mg-1875mg、500mg-2000mg、750mg-1500mg、750mg-1875mg、750mg-2000mg和1875mg-2000mg。
在本发明的第四方面，提供了制备呈药物组合物形式的组合产品的方法。为了提高作为药物的操作性或用于生命体时的吸收性等，优选根据常用方法，将柠檬苦素类化合物或其药学上可接受的衍生物、其酯、其立体异构体、其盐或前药以及磺酰脲类药物或其药学上可接受的衍生物，与药学上可接受的载体、赋形剂、稀释剂等药用辅料进行组合而实现制剂化，从而得到所述形式。
在本发明的第五方面，提供了一种药盒，所述药盒包含本文中所述的组合产品。
本说明书中通篇使用的术语“药学上可接受的盐”是指游离酸或游离碱的盐，通常通过将游离碱与合适的有机或无机酸反应或者通过将酸与合适的有机或无机碱反应而进行制备。该术语可以用于任何化合物，包括柠檬苦素类化合物(具有游离酸或游离碱的功能)等。代表性的盐包括：乙酸盐、苯磺酸盐、苯甲酸盐、碳酸氢盐、硫酸氢盐、酒石酸氢盐、硼酸盐、溴化物、依地酸钙盐、樟脑磺酸盐、碳酸盐、氯化物、克拉维酸盐、柠檬酸盐、二盐酸盐、依地酸盐、乙二磺酸盐、丙酸酯月桂硫酸盐(estolate)、乙磺酸盐(esylate)、富马酸盐、葡庚糖酸盐、葡萄糖酸盐、谷氨酸盐、甘苯砷酸盐(glycol lylarsanilate)、己基间苯二酚酸盐(hexylresorcinate)、哈胺盐(hydrabamine)、氢溴酸盐、盐酸盐、羟萘酸盐、碘化物、羟乙基磺酸盐、乳酸盐、乳糖酸盐、月桂酸盐、苹果酸盐、马来酸盐、扁桃酸盐、甲磺酸盐、甲溴酸盐、甲硝酸盐、甲硫酸盐、马来酸单钾盐、粘酸盐(Mucate)、萘磺酸盐、硝酸盐、N-甲葡萄糖胺盐、草酸盐、巴莫酸盐(双羟萘酸盐)、棕榈酸盐、泛酸盐、磷酸盐/二磷酸盐、聚半乳糖醛酸盐、钾盐、水杨酸盐、钠盐、硬脂酸盐、次乙酸盐、琥珀酸盐、丹宁酸盐、酒石酸盐、茶氯酸盐、对甲苯磺酸盐、三乙基碘盐(triethiodide)、三甲胺盐和戊酸盐。当酸性取代基存在时，例如-COOH，可以形成铵盐、吗啉盐、钠盐、钾盐、钡盐、钙盐等以供剂型使用。当碱性基团存在时，例如氨基或碱性杂芳基如吡啶基，可形成酸性盐，如盐酸盐、氢溴酸盐、磷酸盐、硫酸盐、三氟乙酸盐、三氯乙酸盐、乙酸盐、草酸盐、马来酸盐、丙酮酸盐、丙二酸盐、琥珀酸盐、柠檬酸盐、酒石酸盐、富马酸盐、扁桃酸盐、苯甲酸盐、肉桂酸盐、甲磺酸盐、乙磺酸盐、苦味酸盐等。
本发明中所指磺酰脲类药物的来源，可包括但不限于：格列本脲单独片剂/胶囊、格列齐特单独片剂/胶囊、格列呲嗪单独片剂/胶囊、格列喹酮单独片剂/胶囊、格列美脲单独片剂/胶囊、格列本脲二甲双胍片/胶囊、格列齐特二甲双胍片/胶囊、格列呲嗪二甲双胍片/胶囊、格列喹酮二甲双胍片/胶囊、格列美脲二甲双胍片/胶囊、盐酸吡格列酮格列美脲片、格列美脲马来酸罗格列酮片等等。
实施例
下面通过具体的实施例、对照例进一步说明本发明，但是，应当理解为，这些实施例、对照例仅仅是用于更详细具体地说明之用，而不应理解为用于以任何形式限制本发明。
本发明实施例，选用了下述糖尿病小鼠模型(所述模型是本领域技术人员公知的，或根据本领域常规的教科书、技术手册和科学文献容易获得的)，用以模拟人体不同糖尿病阶段的病理状况。本实施例所提及的柠檬苦素类化合物的存在形式呈单体或提取物的形式。单体是提取得到的或人工合成的，其来源可以商购，或通过本领域现有技术能够容易地制备和得到它们。
实施例1
柠檬苦素类化合物、格列本脲或其组合在小鼠胰岛β细胞损伤模型中对血糖的影响
在本实施例中，用链脲霉素(STZ)造模ICR小鼠得到小鼠胰岛β细胞损伤模型(李楠等，松花粉对糖尿病肾病小鼠肾脏损伤的保护作用，科技导报，2014，32(4/5)：95-99)，完成动物降糖功效评价实验(该模型可以模拟I型、II型糖尿病胰岛β细胞损伤状态)。柠檬苦素类化合物选择柠檬苦素、异柠檬内酯酸、柠檬苦素糖苷、异柠檬内酯酸糖苷，设置单独的格列本脲给药组、柠檬苦素给药组、异柠檬内酯酸给药组、柠檬苦素糖苷给药组、异柠檬内酯酸糖苷给药组以及与格列本脲复配的联合给药组。
实验饲养的条件：ICR小鼠(20±2g)，6周龄，购自浙江省医学科学院，进行7天的预备饲养后进行实验饲养。应予说明，小鼠的饲养条件为温度23±1℃、湿度55±10％、上午7点～下午7点之间亮灯(其他时间关灯)、使小鼠自由摄取水和饲料。实验饲料使用小鼠生长稳定饲料(国标M2118)，动物的日常饲养管理由动物保障部负责，每日为动物提供足够的块料和新鲜的饮用水。
实验分组：取雄性小鼠，随机15只作为正常对照组，其余小鼠禁食12h后，150mg/kg剂量一次性腹腔注射STZ，72h后选取血糖值为15-25mmol/L的小鼠无差异分组纳入实验，每组15只，给药2周后，取血检测指标。
灌胃剂量：柠檬苦素组每日灌胃给予0.02g/kg、异柠檬内酯酸组每日灌胃给予0.02g/kg、柠檬苦素糖苷每日灌胃给予0.02g/kg、异柠檬内酯酸糖苷每日灌胃给予0.02g/kg、格列本脲组每日灌胃给予格列本脲0.02g/kg，柠檬苦素格列本脲联用组每日同时灌胃给予柠檬苦素0.01g/kg以及格列本脲0.01g/kg，异柠檬内酯酸格列本脲联用组每 日同时灌胃给予异柠檬内酯酸0.01g/kg以及格列本脲0.01g/kg，柠檬苦素糖苷格列本脲联用组每日同时灌胃给予柠檬苦素糖苷0.01g/kg以及格列本脲0.01g/kg，异柠檬内酯酸糖苷格列本脲联用组每日同时灌胃给予异柠檬内酯酸糖苷0.01g/kg以及格列本脲0.01g/kg，灌胃量为10mL/kg，正常组和模型组给予10mL/kg蒸馏水。2周后末次给药1h后剪尾测定血糖值(强生血糖仪)，求出各组的平均值。采用SPSS16.0软件进行统计分析，数据以平均值和标准偏差表示，前后数据采用t检验分析，p＜0.05认为有统计学意义。测试结果见下表1。
表1表示每天灌胃给药两周后，STZ小鼠的血糖值
Figure PCTCN2020105728-appb-000002
注*：经独立t检验，与模型组相比，差异极显著(P＜0.05)
**：经独立t检验，与模型组相比，差异极显著(P＜0.01)
实验结果讨论
从上述结果可知，与模型组相比，无论是单独给药还是与格列本脲联用，柠檬苦素及其衍生物均能显著降低STZ胰岛细胞损伤小鼠的血糖值。柠檬苦素及其衍生物与格列本脲联用相对于单独用药效果均有显著提高，显示出协同增效的效果。此外，柠檬苦素及其衍生物与格列本脲联用时，相较单独给药，在降糖效果相当的情况下，可有效降低二者的用药量，提高治疗方案的安全性，降低副作用。
实施例2
柠檬苦素类化合物、格列齐特或其组合在II型糖尿病小鼠模型中对血糖和瘦素的影响
在本实施例中，用db/db小鼠(品系名称BKS.Cg-Dock7 m+/+Lepr db/Nju)完成动物降糖功效评价实验(血糖值和瘦素)。柠檬苦素类化合物选择黄柏酮、异黄柏酮酸和黄柏酮糖苷，分别设置单独的格列齐特给药组、黄柏酮给药组、异黄柏酮酸给药组、黄柏酮糖苷给药组以及分别与格列齐特的联合给药组。
实验饲养的条件：作为II型糖尿病发病模型小鼠，从南京模式生物所购买6周龄的SPF级db/db小鼠，进行7天的预备饲养后进行实验饲养。应予说明，小鼠的饲养条件为温度23±1℃、湿度55±10％、上午7点～下午7点之间亮灯(其他时间关灯)、使小鼠自由摄取水和饲料。实验饲料使用小鼠生长稳定饲料(国标M2118)，动物的日常饲养管理由动物保障部负责，每日为动物提供足够的块料和新鲜的饮用水。
实验分组：选用雄性db/db小鼠(20±2g)，每组18只雄性小鼠进行试验。实验分组为正常对照组(db/m，n＝18)，模型组(db/db，n＝18)，黄柏酮组(db/db，n＝18)，异黄柏酮酸组(db/db，n＝18)，黄柏酮糖苷组(db/db，n＝18)，格列齐特组(db/db，n＝18)，黄柏酮格列齐特联用组(db/db，n＝18)，异黄柏酮酸格列齐特联用组(db/db，n＝18)，黄柏酮糖苷格列齐特联用组(db/db，n＝18)。
灌胃剂量：黄柏酮组每日灌胃给予黄柏酮0.04g/kg、异黄柏酮酸组每日灌胃给予异黄柏酮酸0.04g/kg、黄柏酮糖苷组每日灌胃给予黄柏酮糖苷0.04g/kg、格列齐特组每日灌胃给予格列齐特0.04g/kg，黄柏酮格列齐特联用组每日同时灌胃给予黄柏酮0.02g/kg以及格列齐特0.02g/kg，异黄柏酮酸格列齐特联用组每日同时灌胃给予异黄柏酮酸0.02g/kg以及格列齐特0.02g/kg，黄柏酮糖苷格列齐特联用组每日同时灌胃给予黄柏酮糖苷0.02g/kg以及格列齐特0.02g/kg，灌胃量为10mL/kg，正常组和模型组给予10mL/kg蒸馏 水。2周后末次给药1h后剪尾测定血糖值(强生血糖仪)，并且眼眶内眦取血，酶联免疫吸附法(Elisa)测定血清瘦素水平，求出各组的平均值。采用SPSS16.0软件进行统计分析，数据以平均值和标准偏差表示，前后数据采用t检验分析，p＜0.05认为有统计学意义。测试结果见下表2。
表2表示每天灌胃给药两周后，db/db小鼠的血糖值和瘦素
Figure PCTCN2020105728-appb-000003
注**：经独立t检验，与模型组相比，差异极显著(P＜0.01)
*：经独立t检验，与模型组相比，差异极显著(P＜0.05)
实验结果讨论
从上述结果可知，与模型组相比，无论是单独给药还是与格列齐特联用，黄柏酮及其衍生物均能显著降低db/db糖尿病小鼠的血糖值。黄柏酮及其衍生物与格列齐特联用相对于单独用药效果均有显著提高，显示出协同增效的效果。此外，黄柏酮及其衍生物与格列齐特联用时，相较单独给药，在降糖效果相当的情况下，可有效降低二者的用药量，提高治疗方案的安全性，降低副作用。
同时，黄柏酮及其衍生物为代表的柠檬苦素类化合物能显著提升瘦素敏感性，特别是在联合格列齐特使用时，能明显改善瘦素体内的利用效率，改善机体糖代谢机能，改善糖尿病小鼠糖代谢的相关功能。
实施例3
柠檬苦素类化合物、格列呲嗪或其组合在小鼠胰岛β细胞损伤模型中对血糖的影响
在本实施例中，用链脲霉素(STZ)造模ICR小鼠得到小鼠胰岛β细胞损伤模型(李楠等，松花粉对糖尿病肾病小鼠肾脏损伤的保护作用，科技导报，2014，32(4/5)：95-99)，完成动物降糖功效评价实验(该模型可以模拟I型、II型糖尿病胰岛β细胞损伤状态)。柠檬苦素类化合物选择宜昌橙苦素、宜昌根辛、宜昌橙苦素糖苷，设置单独的格列呲嗪给药组、宜昌橙苦素给药组、宜昌根辛给药组、宜昌橙苦素糖苷给药组以及与格列呲嗪复配的联合给药组。
实验饲养的条件：ICR小鼠(20±2g)，6周龄，购自浙江省医学科学院，进行7天的预备饲养后进行实验饲养。应予说明，小鼠的饲养条件为温度23±1℃、湿度55±10％、上午7点～下午7点之间亮灯(其他时间关灯)、使小鼠自由摄取水和饲料。实验饲料使用小鼠生长稳定饲料(国标M2118)，动物的日常饲养管理由动物保障部负责，每日为动物提供足够的块料和新鲜的饮用水。
实验分组：取雄性小鼠，随机15只作为正常对照组，其余小鼠禁食12h后，150mg/kg剂量一次性腹腔注射STZ，72h后选取血糖值为15-25mmol/L的小鼠无差异分组纳入实验，每组15只，给药2周后，取血检测指标。
灌胃剂量：宜昌橙苦素组每日灌胃给予0.1g/kg、宜昌根辛组每日灌胃给予0.1g/kg、宜昌橙苦素糖苷每日灌胃给予0.1g/kg、格列呲嗪组每日灌胃给予格列呲嗪0.1g/kg，宜昌橙苦素格列呲嗪联用组每日同时灌胃给予宜昌橙苦素0.05g/kg以及格列呲嗪0.05g/kg，宜昌根辛格列呲嗪联用组每日同时灌胃给予宜昌根辛0.05g/kg以及格列呲嗪0.05g/kg，宜昌橙苦素糖苷格列呲嗪联用组每日同时灌胃予宜昌橙苦素糖苷0.05g/kg以及格列呲嗪0.05g/kg，灌胃量为10mL/kg，正常组和模型组给予10mL/kg蒸馏水。2周后末次给药1h后剪尾测定血糖值(强生血糖仪)，求出各组的平均值。采用SPSS16.0软件进行统计分析，数据以平均值和标准偏差表示，前后数据采用t检验分析，p＜0.05认为有统计学意义。测试结果见下表3。
表3表示每天灌胃给药两周后，STZ小鼠的血糖值
Figure PCTCN2020105728-appb-000004
注**：经独立t检验，与模型组相比，差异极显著(P＜0.01)
*：经独立t检验，与模型组相比，差异极显著(P＜0.05)
实验结果讨论
从上述结果可知，与模型组相比，无论是单独给药还是与格列呲嗪联用，上述三种柠檬苦素类化合物均能显著降低STZ胰岛细胞损伤模型小鼠的血糖值。与格列呲嗪联用时，相对于单独用药效果均有显著提高，与正常小鼠血糖值相近，显示出协同增效的效果。此外，上述三种柠檬苦素类化合物与格列呲嗪联用时，相较单独给药，在降糖效果相当的情况下，可有效降低二者的用药量，提高治疗方案的安全性，降低副作用。
实施例4
柠檬苦素类化合物、格列喹酮或其组合在II型糖尿病小鼠模型中对血糖和胰岛素的影响
在本实施例中，柠檬苦素类化合物选择诺米林、脱乙酰诺米林、诺米林酸、脱乙酰诺米林酸糖苷，分别设置单独的格列喹酮给药组、诺米林给药组、脱乙酰诺米林给药组、诺米林酸给药组、脱乙酰诺米林酸糖苷给药组以及分别与格列喹酮联合给药组。
实验饲养的条件：作为II型糖尿病发病模型小鼠，从南京模式生物所购买6周龄的SPF级db/db小鼠，进行7天的预备饲养后进行实验饲养。应予说明，小鼠的饲养条件为温度23±1℃、湿度55±10％、上午7点～下午7点之间亮灯(其他时间关灯)、使小鼠自由摄取水和饲料。实验饲料使用小鼠生长稳定饲料(国标M2118)，动物的日常饲养管理由动物保障部负责，每日为动物提供足够的块料和新鲜的饮用水。
实验分组：选用雄性db/db小鼠(20±2g)，每组18只雄性小鼠进行试验，饮水瓶每周灭菌处理一次。实验分组为正常对照组(db/m，n＝18)，模型组(db/db，n＝18)，诺米林组(db/db，n＝18)，脱乙酰诺米林组(db/db，n＝18)，诺米林酸组(db/db，n＝18)，脱乙酰诺米林酸糖苷组(db/db，n＝18)，格列喹酮组(db/db，n＝18)，诺米林联用组(db/db，n＝18)，脱乙酰诺米林联用组(db/db，n＝18)，诺米林酸联用组(db/db，n＝18)，脱乙酰诺米林酸糖苷联用组(db/db，n＝18)。
灌胃剂量：诺米林组每日灌胃给予诺米林0.2g/kg、脱乙酰诺米林组每日灌胃给予脱乙酰诺米林0.2g/kg、诺米林酸组每日灌胃给予诺米林酸0.2g/kg、脱乙酰诺米林酸糖苷组每日灌胃给予脱乙酰诺米林酸糖苷0.2g/kg、格列喹酮组每日灌胃给予格列喹酮0.2g/kg、诺米林联合用药组每日同时灌胃给予诺米林0.1g/kg以及格列喹酮0.1g/kg，诺米林酸联合用药组每日同时灌胃给予诺米林酸0.1g/kg以及格列喹酮0.1g/kg，脱乙酰诺米林联合用药组每日同时灌胃给予脱乙酰诺米林0.1g/kg以及格列喹酮0.1g/kg，脱乙酰诺米林酸糖苷联合用药组每日同时灌胃给予脱乙酰诺米林酸糖苷0.1g/kg以及格列喹酮0.1g/kg，灌胃量为10mL/kg，正常组和模型组给予10mL/kg蒸馏水。2周后末次给药1h后剪尾测定血糖值(强生血糖仪)，并且眼眶内眦取血，酶联免疫吸附法(Elisa)测定血清胰岛素水平，求出各组的平均值。采用SPSS16.0软件进行统计分析，数据以平均值和标准偏差表示，前后数据采用t检验分析，p＜0.05认为有统计学意义。测试结果见下表4。
表4表示每天灌胃给药两周后，db/db小鼠的血糖值和胰岛素
Figure PCTCN2020105728-appb-000005
注**：经独立t检验，与模型组相比，差异极显著(P＜0.01)
*：经独立t检验，与模型组相比，差异极显著(P＜0.05)
实验结果讨论
从上述结果可知，与模型组相比，无论是单独给药还是与格列喹酮联用，诺米林及其衍生物均能显著降低db/db糖尿病小鼠的血糖值。诺米林及其衍生物与格列喹酮联用相对于单独用药效果均有显著提高，显示出协同增效的效果。此外，诺米林及其衍生物与格列 喹酮联用时，相较单独给药，在降糖效果相当的情况下，可有效降低二者的用药量，提高治疗方案的安全性，降低副作用。
同时，诺米林及其衍生物为代表的柠檬苦素类化合物能显著提升胰岛素敏感性，特别是在联合格列喹酮使用时，能明显改善胰岛素体内的利用效率，改善机体糖代谢机能，改善糖尿病小鼠糖代谢的相关功能。
实施例5
柠檬苦素类化合物、格列美脲或其组合在小鼠II型糖尿病胰岛受损肥胖模型中对血糖的影响
在本实施例中，用链脲霉素(STZ)小剂量多次造模，ICR小鼠连续高脂饮食后，得到小鼠II型糖尿病胰岛受损肥胖模型(参考文献：张继媛等，三种植物提取物对Ⅱ型糖尿病小鼠糖脂代谢改善作用研究，食品与机械，2016，32(12)：142-147)。柠檬苦素类化合物选择诺米林糖苷、脱乙酰基诺米林糖苷、诺米林酸糖苷，设置单独的格列美脲给药组、诺米林糖苷给药组、脱乙酰基诺米林糖苷给药组、诺米林酸糖苷给药组以及与格列美脲复配的联合给药组。
实验饲养的条件：ICR小鼠(20±2g)，6周龄，购自浙江省医学科学院，进行7天的预备饲养后进行实验饲养。应予说明，小鼠的饲养条件为温度23±1℃、湿度55±10％、上午7点～下午7点之间亮灯(其他时间关灯)、使小鼠自由摄取水和饲料。实验饲料使用小鼠生长稳定饲料(国标M2118)，动物的日常饲养管理由动物保障部负责，每日为动物提供足够的块料和新鲜的饮用水。
实验分组：取雄性小鼠，随机15只作为正常对照组，其余小鼠高脂饮食(高脂饲料配方：胆固醇1％、蛋黄粉10％、猪油10％、基础饲料79％以建立肥胖小鼠模型)连续四周，连续三天35mg/kg剂量腹腔注射STZ，1周后禁食禁水24h，测定空腹血糖，选取血糖值为15-25mmol/L的小鼠无差异分组纳入实验，继续高脂饮食，每组15只，给药2周后，取血检测指标。
灌胃剂量：诺米林糖苷组每日灌胃给予0.5g/kg、脱乙酰基诺米林糖苷组每日灌胃给予0.5g/kg、诺米林酸糖苷每日灌胃给予0.5g/kg、格列美脲组每日灌胃给予格列美脲0.5g/kg，诺米林糖苷格列美脲联用组每日同时灌胃给予诺米林糖苷0.25g/kg以及格列美脲0.25g/kg，脱乙酰基诺米林糖苷格列美脲联用组每日同时灌胃给予脱乙酰基诺米林糖苷 0.25g/kg以及格列美脲0.25g/kg，诺米林酸糖苷格列美脲联用组每日同时灌胃给予诺米林酸糖苷0.25g/kg以及格列美脲0.25g/kg，灌胃量为10mL/kg，正常组和模型组给予10mL/kg蒸馏水。2周后末次给药1h后剪尾测定血糖值(强生血糖仪)，求出各组的平均值。采用SPSS16.0软件进行统计分析，数据以平均值和标准偏差表示，前后数据采用t检验分析，p＜0.05认为有统计学意义。测试结果见下表5。
表5表示每天灌胃给药两周后，STZ小鼠的血糖值
Figure PCTCN2020105728-appb-000006
注**：经独立t检验，与模型组相比，差异极显著(P＜0.01)
*：经独立t检验，与模型组相比，差异极显著(P＜0.05)
实验结果讨论
从上述结果可知，与模型组相比，无论是单独给药还是与格列美脲联用，上述三种柠檬苦素类糖苷均能显著降低STZ II型糖尿病模型小鼠的血糖值。与格列美脲联用时，相对 于单独用药效果均有显著提高，与正常小鼠血糖值相近，显示出协同增效的效果。此外，上述三种柠檬苦素类糖苷与格列美脲联用时，相较单独给药，在降糖效果相当的情况下，可有效降低二者的用药量，提高治疗方案的安全性，降低副作用。
实施例6
含有诺米林和格列本脲的组合产品的片剂的制备方法
在本实施例中，示例性提供了本发明的组合产品(诺米林和格列本脲)的片剂的制备方法。单个片剂含有下述成分：诺米林50mg，盐酸格列本脲400mg，羟丙基甲基纤维素的用量为20mg，羧甲基纤维素钠的用量为30mg，微晶纤维素的用量为20mg，硬脂酸镁5.2mg，欧巴代20.8mg，共计1000片。
所述制备方法包括以下步骤：
a)将50g诺米林溶解于5L 50％乙醇中；
b)将原、辅料分别过100目筛，备用；
c)称取400g盐酸格列本脲、20g羟丙基甲基纤维素、30g羧甲基纤维素钠、20g微晶纤维素置流化床中，设置进风风量500±50m 3/h、进风温度90±5℃、产品温度70±5℃，进行热熔制粒；
d)将诺米林溶液喷入流化床中，设置雾化压力1.0±0.2bar、喷液速度30±10Hz，进行一步制粒；
e)将制好的颗粒过1.0mm圆孔筛网进行干整粒；
f)加入5.2g硬脂酸镁，混合5min；
g)用17×8.5mm椭圆形冲压片，压力为15KN；
h)用20.8g欧巴代85F32004以1:4溶于蒸馏水中,包衣锅设置参数为：片床温度40±2℃，出风温度48±2℃，雾化压力0.6Mpa，锅速7rpm，喷量120g/min。完成包薄膜衣。

Claims (10)

  1. 一种组合产品，所述组合产品包含柠檬苦素类化合物或其药学上可接受的衍生物、其酯、其立体异构体、其盐或前药，以及磺酰脲类药物或其药学上可接受的衍生物。
  2. 根据权利要求1所述的组合产品，其中所述组合产品呈药物组合物的形式。
  3. 根据权利要求1所述的组合产品，其中所述柠檬苦素类化合物或其药学上可接受的衍生物、其酯、其立体异构体、其盐或前药，以及磺酰脲类药物或其药学上可接受的衍生物各自呈单独的制剂形式。
  4. 根据权利要求3所述的组合产品，其中所述柠檬苦素类化合物或其药学上可接受的衍生物、其酯、其立体异构体、其盐或前药，以及磺酰脲类药物或其药学上可接受的衍生物同时或先后给药。
  5. 根据权利要求1-4中任一项所述的组合产品，其中所述磺酰脲类药物的量是50mg-2000mg。
  6. 根据权利要求1-4中任一项所述的组合产品，其中所述柠檬苦素类化合物的量是50mg-2000mg。
  7. 根据权利要求1-6中任一项的所述的组合产品，其中所述磺酰脲类药物选自格列本脲、格列齐特、格列呲嗪、格列喹酮和格列美脲等，且所述柠檬苦素类化合物选自下述的一个或多个：柠檬苦素、异柠檬内酯酸、7α-柠檬苦醇、黄柏酮、宜昌橙苦素、宜昌根辛、诺米林、脱乙酰诺米林、诺米林酸、脱乙酰诺米林酸、枸橼苦素、异黄柏酮酸及其任何的糖苷衍生物。
  8. 根据权利要求7所述的组合产品，所述组合产品还包含药学上可接受的载体、稀释剂或赋形剂。
  9. 根据权利要求8所述的组合产品，其中所述组合产品呈片剂、胶囊剂、颗粒剂、糖浆剂、粉剂、锭剂、药囊、扁囊剂、酏剂、混悬剂、乳剂、溶液、糖浆剂、气雾剂、软膏剂、乳膏剂和注射剂的形式。
  10. 根据权利要求1-9中任一项的组合产品在制备药物中的用途，所述药物用于预防和/或治疗糖尿病以及代谢综合症相关疾病。
PCT/CN2020/105728 2019-08-12 2020-07-30 包含柠檬苦素类化合物和磺酰脲类药物的组合产品 WO2021027583A1 (zh)

Priority Applications (3)

Application Number Priority Date Filing Date Title
JP2022508933A JP7383126B2 (ja) 2019-08-12 2020-07-30 リモノイド化合物およびスルホニル尿素化合物を含有する組合せ製品
EP20853232.5A EP4014999B1 (en) 2019-08-12 2020-07-30 Combination product containing limonin compound and sulfonylurea drug
US17/633,953 US20220296615A1 (en) 2019-08-12 2020-07-30 Combination product containing limonoid compound and sulfonylurea compound

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201910741989.4A CN110302386B (zh) 2019-08-12 2019-08-12 包含柠檬苦素类化合物和磺酰脲类药物的组合产品
CN201910741989.4 2019-08-12

Publications (1)

Publication Number Publication Date
WO2021027583A1 true WO2021027583A1 (zh) 2021-02-18

Family

ID=68083492

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2020/105728 WO2021027583A1 (zh) 2019-08-12 2020-07-30 包含柠檬苦素类化合物和磺酰脲类药物的组合产品

Country Status (6)

Country Link
US (1) US20220296615A1 (zh)
EP (1) EP4014999B1 (zh)
JP (1) JP7383126B2 (zh)
CN (1) CN110302386B (zh)
TW (1) TWI747406B (zh)
WO (1) WO2021027583A1 (zh)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110302386B (zh) * 2019-08-12 2021-05-04 浙江养生堂天然药物研究所有限公司 包含柠檬苦素类化合物和磺酰脲类药物的组合产品
CN110279866B (zh) * 2019-08-12 2021-06-29 浙江养生堂天然药物研究所有限公司 包含柠檬苦素类化合物和噻唑烷二酮类药物的组合产品

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102481276A (zh) * 2009-04-20 2012-05-30 埃尔舍利克斯治疗公司 基于化学感受受体配体的治疗法
CN103597071A (zh) * 2011-01-07 2014-02-19 埃尔舍利克斯治疗公司 基于化学感应受体配体的治疗
US20140371303A1 (en) * 2013-06-17 2014-12-18 Kgk Synergize, Inc. Compositions and methods for glycemic control of subjects with impaired fasting glucose
CN109381471A (zh) * 2017-08-08 2019-02-26 浙江养生堂天然药物研究所有限公司 包含柠檬苦素类化合物和双胍类化合物的组合产品
CN110302386A (zh) * 2019-08-12 2019-10-08 浙江养生堂天然药物研究所有限公司 包含柠檬苦素类化合物和磺酰脲类药物的组合产品

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2000072684A (ja) * 1998-08-26 2000-03-07 Miyazaki Prefecture 血清脂質代謝改善剤
US8309080B2 (en) * 2007-12-06 2012-11-13 Naidu Lp Metallo-protein and tocotrienol (MP-T3) compositions and uses thereof
CN103446385B (zh) * 2013-08-26 2015-07-08 山西康禾农业有限责任公司 一种治疗糖尿病并发症的中药制剂
CN106132425B (zh) * 2014-02-14 2020-09-11 龟甲万株式会社 包含柑橘类种子提取物的组合物、食品、药品和包含柑橘类种子提取物的组合物制造方法
CN108210600A (zh) * 2017-12-27 2018-06-29 济南昊雨青田医药技术有限公司 一种柠檬苦素类提取物的制备方法及其应用
CN108210566A (zh) * 2017-12-27 2018-06-29 济南昊雨青田医药技术有限公司 一种柠檬苦素类提取物在制备治疗糖尿病药物中的应用

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102481276A (zh) * 2009-04-20 2012-05-30 埃尔舍利克斯治疗公司 基于化学感受受体配体的治疗法
CN103597071A (zh) * 2011-01-07 2014-02-19 埃尔舍利克斯治疗公司 基于化学感应受体配体的治疗
US20140371303A1 (en) * 2013-06-17 2014-12-18 Kgk Synergize, Inc. Compositions and methods for glycemic control of subjects with impaired fasting glucose
CN109381471A (zh) * 2017-08-08 2019-02-26 浙江养生堂天然药物研究所有限公司 包含柠檬苦素类化合物和双胍类化合物的组合产品
CN110302386A (zh) * 2019-08-12 2019-10-08 浙江养生堂天然药物研究所有限公司 包含柠檬苦素类化合物和磺酰脲类药物的组合产品

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
GUALDANI ROBERTA, CAVALLUZZI MARIA, LENTINI GIOVANNI, HABTEMARIAM SOLOMON: "The Chemistry and Pharmacology of Citrus Limonoids", MOLECULES, vol. 21, no. 11, pages 1530, XP055780471, DOI: 10.3390/molecules21111530 *
LI NAN ET AL.: "Protective effect of pine pollen on kidney damage in diabetic nephropathy mice", SCIENCE AND TECHNOLOGY REVIEW, vol. 32, no. 4/5, 2014, pages 95 - 99
S. A. MOHAMMED, A. G. YAQUB, K. A. SANDA, A. O. NICHOLAS, W. ARASTUS, M. MUHAMMAD AND S. ABDULLAHI: "Review on diabetes, synthetic drugs and glycemic effects of medicinal plants", JOURNAL OF MEDICINAL PLANTS RESEARCH, vol. 7, no. 36, 25 September 2013 (2013-09-25), pages 2628 - 2637, XP009526005, DOI: 10.5897/JMPR2013.5169 *
See also references of EP4014999A4
YAN MIN, ZHOU YU; HE XIAO-HAN; MEI MING-XIN; DONG QUAN: "FOOD AND FEï¼²MENTATION INDUSTï¼²IES", FOOD AND FERMENTATION INDUSTRIES, vol. 44, no. 2, 1 January 2018 (2018-01-01), pages 290 - 296, XP055780486, DOI: 10.13995/j.cnki.11-1802/ts.014827 *
ZHANG JIYUAN ET AL.: "Study on the effect of three plant extracts on improving glucose and lipid metabolism in type 2 diabetic mice", FOOD AND MACHINERY, vol. 32, no. 12, 2016, pages 142 - 147

Also Published As

Publication number Publication date
TWI747406B (zh) 2021-11-21
EP4014999B1 (en) 2023-11-29
CN110302386B (zh) 2021-05-04
EP4014999A4 (en) 2022-11-23
EP4014999A1 (en) 2022-06-22
CN110302386A (zh) 2019-10-08
US20220296615A1 (en) 2022-09-22
TW202120097A (zh) 2021-06-01
JP7383126B2 (ja) 2023-11-17
JP2022544517A (ja) 2022-10-19

Similar Documents

Publication Publication Date Title
US11679097B2 (en) Combination product containing limonoid compound and biguanide compound
TWI747406B (zh) 包含檸檬苦素類化合物和磺醯尿素類藥物的組合產品及其用途
TWI782301B (zh) 包含檸檬苦素類化合物和dpp-4抑制劑的組合產品
JP7398547B2 (ja) リモノイド化合物およびα－グルコシダーゼ阻害剤を含有する組合せ製品
TWI785357B (zh) 包含檸檬苦素類化合物和噻唑烷二酮類藥物的組合產品及其用途
TWI779326B (zh) 包含檸檬苦素類化合物和sglt-2抑制劑的組合產品

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 20853232

Country of ref document: EP

Kind code of ref document: A1

ENP Entry into the national phase

Ref document number: 2022508933

Country of ref document: JP

Kind code of ref document: A

NENP Non-entry into the national phase

Ref country code: DE

ENP Entry into the national phase

Ref document number: 2020853232

Country of ref document: EP

Effective date: 20220314