Nothing Special   »   [go: up one dir, main page]

WO2016180353A1 - 一种包含glp-1类似物的药物制剂及其制备方法 - Google Patents

一种包含glp-1类似物的药物制剂及其制备方法 Download PDF

Info

Publication number
WO2016180353A1
WO2016180353A1 PCT/CN2016/081843 CN2016081843W WO2016180353A1 WO 2016180353 A1 WO2016180353 A1 WO 2016180353A1 CN 2016081843 W CN2016081843 W CN 2016081843W WO 2016180353 A1 WO2016180353 A1 WO 2016180353A1
Authority
WO
WIPO (PCT)
Prior art keywords
glp
poloxamer
pharmaceutical composition
amide
val
Prior art date
Application number
PCT/CN2016/081843
Other languages
English (en)
French (fr)
Inventor
刘晓妮
李寒星
陈潮梁
马国昌
徐飞虎
王同映
孙汉栋
Original Assignee
杭州九源基因工程有限公司
刘晓妮
李寒星
陈潮梁
马国昌
徐飞虎
王同映
孙汉栋
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 杭州九源基因工程有限公司, 刘晓妮, 李寒星, 陈潮梁, 马国昌, 徐飞虎, 王同映, 孙汉栋 filed Critical 杭州九源基因工程有限公司
Priority to EP16792203.8A priority Critical patent/EP3295952B1/en
Priority to US15/571,278 priority patent/US20190060410A1/en
Priority to CN201680007374.3A priority patent/CN107249620B/zh
Publication of WO2016180353A1 publication Critical patent/WO2016180353A1/zh

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/22—Hormones
    • A61K38/26—Glucagons
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/02—Inorganic compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
    • A61K47/10—Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
    • 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
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • 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
    • A61K9/08—Solutions
    • 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 relates to the field of polypeptide pharmaceutical preparations, and more particularly to a pharmaceutical preparation comprising a GLP-1 analogue and a process for the preparation thereof.
  • incretin is an enterogenous hormone in the human body. After eating, this hormone can promote insulin secretion and exert glucose concentration-dependent hypoglycemic effect.
  • Incretin is mainly composed of glucagon-1 (GLP-1) and glycoprotein-dependent insulin releasing peptide (GIP), and GLP-1 plays a more important role in the development of type 2 diabetes.
  • GLP-1 can lower blood sugar by stimulating insulin secretion, inhibiting glucagon secretion and gastric emptying; on the other hand, GLP-1 also has a unique effect of slowing down beta cell apoptosis and promoting its regeneration.
  • GLP-1 when analyzing the gene sequence of the glucagon precursor, proglucagon (PG).
  • the GLP-1 gene is expressed in pancreatic alpha cells and intestinal L cells.
  • the complete GLP-1 polypeptide consists of 37 amino acids, and its 1-37 peptide sequence structure is:
  • GLP-1 (7-37) polypeptide and GLP-1 (7-36) amide of which about 80% of the GLP-1 circulating activity is derived from GLP-1 (7-36). Amide.
  • the amino terminus of GLP-1(7-37)OH is designated as No. 7, and the carboxyl group The end is designated as number 37.
  • GLP-1(1-37) is produced in vivo, it is removed by two-step digestion to remove N-terminal 6 amino acids and form C-terminal amidation, which finally produces highly active GLP-1(7-36) amide (also Called GLP-1 fragment).
  • GLP-1 is the most potent intestinal peptide hormone that has been found to act by binding to the GLP-1 receptor (GLP-1R, a G-coupled protein belonging to the ⁇ receptor family). Upon binding of GLP-1R, GLP-1 activates the intracellular membrane cyclic adenosine monophosphate (cAMP) and mitogen-activated protein kinase (MAPK) pathways.
  • cAMP membrane cyclic adenosine monophosphate
  • MAPK mitogen-activated protein kinase
  • pancreatic islet mature ⁇ cells is coupled with Gs, activates adenylyl cyclase, and produces cAMP, which synergizes with glucose to stimulate insulin synthesis and secretion, stimulates insulin gene transcription and proinsulin biosynthesis, and reduces pancreatic hyperglycemia.
  • the concentration of the hormone inhibits the secretion of glucagon, enhances the sensitivity of the cells to insulin, stimulates insulin-dependent glycogen synthesis, and reduces postprandial blood glucose concentration.
  • PI3K protein kinase
  • MAPK channels By activating protein kinase, phosphatidylinositol 3-kinase (PI3K) and MAPK channels, it regulates pre-apoptotic proteins and induces the expression of anti-apoptotic proteins Bcl-2 and Bcl-xL to slow down ⁇ -cell apoptosis and enhance its regeneration.
  • Islet beta cells differentiate and proliferate.
  • GLP-1 can slow down the rate of gastric emptying; by appetizing the hypothalamus, appetite is suppressed.
  • polypeptides such as GLP-1 and its analogs are mostly prepared by genetic recombination. Recombinant preparation of proteins and polypeptides often uses E. coli and yeast as hosts. Among them, the drug that has been marketed and dominates the mainstream market of GLP-1 analogue drugs is liraglutide, trade name Victoza. Liraglutide chemical name Arg 34 , Lys 26 (N ⁇ - ( ⁇ -Glu(N ⁇ -hexadecanoyl))) GLP-1 (7-37), is a fatty acid modified GLP-1 polypeptide The analog, the modified drug, was extended to a plasma half-life of 13 hours after subcutaneous administration.
  • the GLP-1 polypeptide chain sequence is similar to the human GLP-1 (7-37) sequence, and the human GLP-1 native sequence 34th lysine is substituted with arginine, and its amino acid sequence is associated with glucagon-like glucagon-like
  • the peptide-1 (GLP-1) has about 97% homology (Goke et al, J. Biol. Chem., 268: 19650-55, 1993).
  • the 16-carbon fatty acid is linked by a glutamic acid and a 26th lysine of the polypeptide, and its structure is as follows:
  • Liraglutide as a GLP-1 analogue, has the following pharmacological effects due to its high homology to GLP-1: (1) blood glucose concentration-dependent insulin secretion; (2) inhibition of postprandial glucagon secretion; (3) Reduce appetite and slow gastric emptying.
  • liraglutide can promote B cell proliferation and differentiation, regulate the expression of B cell apoptosis gene, inhibit its apoptosis, and increase the number of islet B cells in vivo. Studies have shown that liraglutide can also restore the sensitivity of human islet B cells to blood glucose.
  • the first line of drugs commonly used to treat type 2 diabetes is oral hypoglycemic agents, but the adverse effects of current oral hypoglycemic agents, especially hypoglycemia, make it unsuitable for all patients with type 2 diabetes.
  • the LEAD study included six phase III clinical trials that evaluated the efficacy of liraglutide monotherapy and other oral hypoglycemic agents.
  • HbA1c glycated hemoglobin
  • liraglutide has a protective effect on the cardiovascular system. Studies have found that liraglutide can reduce systolic blood pressure by 2.7 ⁇ 6.7mmHg, which occurs under the weight of patients Before the drop, the decline in systolic blood pressure cannot be completely attributed to changes in the patient's body weight. The study found that patients receiving liraglutide have a slightly increased pulse rate, and the clinical significance of this phenomenon remains to be further studied. Courreges et al reported that liraglutide treatment was accompanied by a decrease in the concentrations of total cholesterol, low density lipoprotein, free fatty acids, triglycerides, plasminogen activator inhibitor-1 and type B natriuretic peptide. The study also found that high-sensitivity C-reactive protein levels in the liraglutide-treated group showed a dose-dependent decrease, albeit not significantly.
  • Liraglutide a GLP-1 analogue
  • causes hypoglycemia Clinical trials have shown that the incidence of hypoglycemia in liraglutide monotherapy is significantly lower than that of glimepiride, which is comparable to metformin.
  • the hypoglycemic event caused by liraglutide combination therapy is comparable to or less than other hypoglycemic agents.
  • the most common adverse reaction in the treatment of liraglutide is gastrointestinal reactions, mainly nausea, vomiting, and diarrhea, usually seen in the first week of administration, in a dose-dependent manner.
  • gastrointestinal reactions mainly nausea, vomiting, and diarrhea, usually seen in the first week of administration, in a dose-dependent manner.
  • patients with nausea in the liraglutide-treated group were ⁇ 10%.
  • the gastrointestinal reaction can be reduced by slowly increasing the dose. Patients often tolerate this adverse reaction within 1 to 3 weeks of treatment, and few patients discontinue treatment due to gastrointestinal reactions.
  • liraglutide can rapidly and efficiently reduce blood glucose and HbA1c levels, either alone or in combination with other oral hypoglycemic agents. Its hypoglycemic effect depends on the glucose concentration, that is, the insulin release is stimulated to achieve a therapeutic effect only when the blood glucose level is elevated, so the probability of occurrence of hypoglycemia is very low. Most striking is its potential to delay the progression of diabetes. A large number of clinical trials have shown that liraglutide can improve B cell function, reduce its apoptosis, increase the regeneration of B cells, and thus delay the progression of diabetes. The protective effect of liraglutide on the cardiovascular system can reduce the incidence of cardiovascular complications in diabetes.
  • liraglutide has a significant weight-reducing effect and is suitable for patients who need to lose weight and have a risk of severe hypoglycemia.
  • the unique pharmacological effects of liraglutide make it a promising future in the treatment of diabetes.
  • GLP-1 protein is a kind of protein which is more easily degraded, and many hydrolase enzymes are easy to degrade (M.Egel-Mitani, et al; Yield improvement of heterologous peptides expressed in yps1-disrupted Saccharomyces cerevisiae strains Enzyme and Microbial Technology, 2000, 26: 671-677).
  • liraglutide is more stable after fatty acid chain modification, its stability as a peptide drug cannot be compared with conventional chemical drugs, and its physical and chemical properties are subject to various environmental factors during long-term storage.
  • the formation of foreign matter in the formulation solution is usually caused by the interaction of physical oscillations with protein molecules in solution and the interaction of protein molecules at the liquid-vapor interface in the storage vial. It is believed that protein molecules are adsorbed at the gas-liquid interface, with hydrophobic groups extending into the air and hydrophilic groups immersed in the aqueous phase. Once arranged on the surface, protein molecules tend to aggregate, form particles and precipitate. It is also believed that in the formulation of the stretching process of the interface, such as due to vibrations occurring during transportation or other processes, proteins adsorbed to the gas-liquid and solid-liquid interface undergo further conformational changes. This oscillation can cause protein to entangle, aggregate, form particles and eventually precipitate with other adsorbed proteins. In addition, during the lyophilization process, the protein is exposed to foreign matter due to the effects of freeze-drying curve processes such as pre-freezing rate and heating rate.
  • CN200480034152.8 Chinese patent application mentions an improved formulation of liraglutide.
  • liraglutide is easily crystallized from the drug solution, for example, a preparation containing mannitol causes clogging due to crystallization of the production equipment or the syringe, thereby affecting the production control of the device or the treatment of the patient.
  • liraglutide drugs are used to treat diabetes, lifelong administration is required and injections are made daily.
  • the marketing prescription of liraglutide is: 3 ml of solution containing 18 mg of liraglutide, 1.42 mg of disodium hydrogen phosphate dihydrate, 14 mg of propylene glycol, 5.5 mg of phenol, dissolved in water for injection, and having a pH of 8.15.
  • propylene glycol is a relatively safe accessory, according to clinical conclusions, propylene glycol still has some possible side effects on human skin, mainly the following:
  • Irritability Some people have a subjective burning sensation, tingling and itching when used.
  • Propylene glycol has the characteristics of a fat-soluble solvent, and long-term use of a high concentration of propylene glycol has an effect on the structure of the epidermal sebum.
  • propylene glycol is irritating to the skin and mucous membranes. The higher the concentration and the more sealed, the more irritating, causing redness, rash, peeling and rough skin. . Although it is not very irritating to most people, the cumulative effect that can be generated by long-term use is very noteworthy. For injured skin or sensitive skin, even a low concentration of propylene glycol is prone to irritating reactions.
  • Allergic dermatitis About 1 to 5% of people are exposed to propylene glycol, which produces a local skin allergic eczema reaction. Usually no problem occurs when you first come into contact, but when you come into contact again, it will cause an allergic reaction to the skin.
  • Systemic contact dermatitis A small number of people who have a skin allergic reaction to propylene glycol may cause systemic skin allergic reactions if they take or apply food or medicine containing propylene glycol.
  • propylene glycol is widely used and safe, but it still has potential irritancy and sensitization to the skin. For its common use, if there are other safer alternatives, minimize the use of propylene glycol or reduce its concentration on the skin. It will be safer.
  • the inventors unexpectedly obtained a stable pharmaceutical composition comprising a GLP-1 analogue by a large number of prescription screening and stability tests.
  • the pharmaceutical composition prepared by the invention comprises 0.1 mg/ml to 25 mg/ml of GLP-1 analogue, pH 7.5-9.0 buffer, 0.001% to 0.05% (m/v) stabilizer, 0.5 %-10% (m/v) of xylitol and 0.1 mg/ml to 10 mg/ml of preservative.
  • GLP-1 analog means GLP-1 or a mutant thereof, GLP-1 (7-36)-amide or a mutant thereof, GLP-1 (7-37) or a mutant thereof And a derivative in which one or more amino acid residues of the above polypeptide or a mutant thereof are chemically modified with an organic substituent such as an ester, an alkyl group or a lipophilic functional group.
  • mutants are that the parent peptide GLP-1, one or more amino acid residues of GLP-1 (7-36)-amide or GLP-1 (7-37) have been substituted by other amino acid residues and/or the parent thereof One or more amino acid residues of the peptide have been deleted and/or one or more amino acid residues have been added to the parent peptide. Such addition can occur at the N-terminus or C-terminus or both ends of the parent peptide.
  • the GLP-1 derivative preferably has three linkages to the parent peptide (ie, GLP-1(7-36)-amide, GLP-1 (7-37), GLP-1 (7) -36)-amide analog Or a lipophilic substituent of the GLP-1 (7-37) analog), more preferably two lipophilic substituents, and most preferably a lipophilic substituent, wherein each lipophilic substituent preferably has 4 to 40 carbons
  • the atom is more preferably 8 to 30 carbon atoms, still more preferably 8 to 25 carbon atoms, still more preferably 12 to 25 carbon atoms, and most preferably 14 to 18 carbon atoms.
  • GLP-1 analogs suitable for the present invention are described in several places in the prior art, and are described in particular in WO 93/19175 (Novo Nordisk), WO 99/43705 (Novo Nordisk) WO 99/43706 (Novo Nordisk), WO 99/ 43707 (Novo Nordisk), WO 98/08871 (an analog having a lipophilic substituent) and WO 02/46227 (an analog fused to serum albumin or to the Fc portion of Ig) (Novo Nordisk A/S), WO 99/43708 (Novo Nordisk A/S), WO 99/43341 (Novo Nordisk A/S), WO 87/06941 (The General Hospital Corporation), WO 90/11296 (The General Hospital Corporation), WO 91/11457 (Buckley et al), WO 98/43658 (Eli Lilly & Co.), EP 0 708 179-A2 (Eli Lilly & Co.), EP 0 699 686-A2 (Eli Li
  • the GLP-1 analogue is preferably Arg 34 , Lys 26 (N ⁇ -( ⁇ -Glu(N ⁇ -hexadecanoyl)))-GLP-1 (7-37), Liraglutide.
  • the GLP-1 analogue is selected from the group consisting of
  • Gly 8 -GLP-1(7-36)-amide Gly 8 -GLP-1 (7-37), Val 8 -GLP-1(7-36)-amide, Val 8 -GLP-1 (7-37 ), Val 8 Asp 22 -GLP-1(7-36)-amide,
  • the GLP-1 analogue is selected from the group consisting of Arg 26 -GLP-1 (7-37);
  • the GLP-1 analogue is selected from
  • the concentration of the GLP-1 analog is preferably from about 1 mg/ml to 15 mg/ml, more preferably from 3 mg/ml to 10 mg/ml, and most preferably 6 mg/ml.
  • the buffering agent suitable for use in the present invention is any buffer capable of maintaining the pH of the preparation in an aqueous solution state of 7.5-9.0, and may be selected from phosphoric acid.
  • Buffer disodium hydrogen phosphate-citrate buffer, TRIS, glycyl-glycine, N-bis(hydroxyethyl)glycine, sodium dihydrogen phosphate buffer, disodium hydrogen phosphate buffer, sodium acetate buffer, Sodium carbonate buffer, sodium phosphate buffer, lysine buffer, arginine buffer or a mixture thereof.
  • the pH of the buffer preferably ranges from 7.5 to 8.5, more preferably from 8.0 to 8.5; the concentration of the buffer is from 5 to 100 mmol/L, preferably from 10 to 30 mmol/L.
  • the disodium hydrogen phosphate buffer solution has a concentration of 5-100 mmol/L and a pH range of 7.5-8.5; more preferably, the concentration of the disodium hydrogen phosphate buffer is 10-30 mmol/L, and the pH ranges from 8.0 to 8.5.
  • xylitol is used as an isotonicity adjusting agent in a weight-to-volume ratio of 0.5% to 10% (m/v), more preferably 1% to 5% (m/v).
  • stabilizers for increasing the stability of GLP-1 analogs include, but are not limited to, amino acids and derivatives: glycine, alanine, serine, aspartic acid, glutamic acid, Threonine, tryptophan, lysine, hydroxylysine, histidine, arginine, cystine, cysteine, methionine, phenylalanine, leucine, isoluminescence Amino acids and their derivatives; nonionic surfactants: sorbitan fatty acid esters, glycerol fatty acid esters (such as sorbitan caprylate monoester, sorbitan lauric acid monoester and sorbitan palm Acid monoester), polyglycerol fatty acid ester (such as glyceryl octanoate monoester, glycerin myristate monoester cream and glycerol hard fatty acid monoester), polyoxyethylene sorbitan fatty acid
  • the stabilizers are preferably polysorbate 20, polysorbate 80, poloxamer 124, poloxamer 181, poloxamer 182, poloxamer 188, poloxamer 237, poloxamer. 331, poloxamer 338, poloxamer 407 or mixtures thereof, more preferably polysorbate 20, polysorbate 80, poloxamer 188 or mixtures thereof.
  • Suitable pharmaceutically acceptable preservatives are included in the pharmaceutical combinations disclosed herein. Suitable pharmaceutically acceptable preservatives may be selected from the group consisting of phenol, o-cresol, m-cresol, p-cresol, methyl p-hydroxybenzoate, propyl p-hydroxybenzoate, 2-phenoxyethanol, p-hydroxybenzene Butyl formate, 2-phenylethanol, benzyl alcohol, chlorobutanol, chlorocresol, ethyl p-hydroxybenzoate or a mixture thereof, at a concentration of about 0.1 mg/ml to 10 mg/ml, preferably at a concentration of about 1 mg/ml. To 8 mg/ml, the most preferred concentration is from about 2 mg/ml to 6 mg/ml.
  • the invention also discloses a preparation method of a GLP-1 analogue pharmaceutical composition, the method comprising the following steps:
  • GLP-1 analogue containing 0.1 mg/ml to 25 mg/ml, a buffer of pH 7.5-9.0, and a stabilizer of 0.001% to 0.5% (m/v).
  • preservatives buffers, stabilizers and GLP-1 analogues are as previously described.
  • compositions disclosed herein are particularly useful for the long-term preservation of liraglutide pharmaceutical formulations.
  • Liraglutide is administered by subcutaneous injection, and in order to stably store the liraglutide preparation, which is not easily degraded and crystallized from the preparation solution, the inventors have found that it is necessary to add a suitable amount of the surfactant to the preparation solution.
  • the type and characteristics of surfactants that are incorporated into parenteral compositions for injection into the human body are limited for reasons of safety for clinical applications. Accordingly, there is a need in the art for pharmaceutical compositions that provide improved protein stability that include only those commercial parenteral ingredients that are considered safe and include in authoritatively approved regulations.
  • the present invention has unexpectedly discovered through the use of a large number of exploration experiments, using xylitol as an isotonic regulator to replace propylene glycol with low concentration of poloxamer surfactant and polysorbate surface activity
  • the agent preferably Tween-20 or Tween-80
  • Tween-20 or Tween-80 alone or in combination can improve the long-term stability of the liraglutide preparation. After storage at 4 ° C for 3 months, the purity of liraglutide remains above 95%. It is more stable than the imported Victoza market prescription; and the formulation solution remains clear, no visible foreign matter or crystallization causes the injection tube to be blocked; moreover, the clinical application of xylitol in diabetes is safer than propylene glycol. More, fewer side effects.
  • xylitol combined with low concentrations of poloxamer surfactants and polysorbate surfactants can reduce the formation of high molecular impurities such as dimers during long-term storage of liraglutide preparations.
  • An unexpected technical effect has never been reported in the prior art literature.
  • Xylitol has a very low glycemic index and is a natural stabilizer for insulin. Its metabolism is independent of insulin, and it does not cause an increase in blood sugar levels. It can eliminate more than three people with diabetes (drinking, polyuria). And eat more), so it is a safe sweetener, nutritional supplement and adjuvant therapeutic for diabetics.
  • xylitol has a high degree of chemical stability, which means it does not interact with drugs and other excipients and can be used over a wide pH range (pH 1-11).
  • xylitol has a lower activity in water and a higher osmotic pressure, thus increasing the stability of the product.
  • the literature reports that xylitol can enhance the preservative effect, and at the same time it has the bacteriostatic and bactericidal properties, which can enhance the stability of the product.
  • the invention also discloses a preparation method of a liraglutide pharmaceutical composition, the method comprising the following steps:
  • the pharmaceutical composition prepared by the invention comprises 0.1 mg/ml to 25 mg/ml of liraglutide, a buffer of pH 7.5-9.0, a stabilizer of 0.001% to 0.05% (m/v), 0.5%. -10% (m/v) Xylitol and a preservative from 0.1 mg/ml to 10 mg/ml.
  • the invention has the advantages that the physicochemical and biological activity of liraglutide is stabilized by adding some components which can be accepted by the human body, thereby preparing a preparation suitable for clinical multiple injection.
  • the preparation can effectively prevent the formation of high molecular weight polymer due to various factors such as degradation, oxidation and precipitation of the active ingredient liraglutide, and the purity is reduced or crystallized, thereby facilitating convenient transportation, long-term preservation and clinical use.
  • the concentration of liraglutide is preferably from about 1 mg/ml to 15 mg/ml, more preferably from 3 mg/ml to 10 mg/ml, and most preferably 6 mg/ml.
  • the buffer suitable for use in the present invention is any buffer capable of maintaining the pH of the preparation in the aqueous solution state of 7.5-9.0, and may be selected from the group consisting of phosphate buffer, disodium hydrogen phosphate-citric acid. Buffer, TRIS, glycyl-glycine, N-bis(hydroxyethyl)glycine, sodium dihydrogen phosphate buffer, disodium hydrogen phosphate buffer, sodium acetate buffer, sodium carbonate buffer, sodium phosphate buffer, Lysine buffer, arginine buffer or a mixture thereof.
  • the pH of the buffer preferably ranges from 7.5 to 8.5, more preferably from 8.0 to 8.5; the concentration of the buffer is from 5 to 100 mmol/L, preferably from 10 to 30 mmol/L.
  • the disodium hydrogen phosphate buffer solution has a concentration of 5-100 mmol/L and a pH range of 7.5-8.5; more preferably, the concentration of the disodium hydrogen phosphate buffer is 10-30 mmol/L, and the pH ranges from 8.0 to 8.5.
  • Xylitol is used as an isotonicity adjusting agent in the pharmaceutical composition of the present invention, and the weight-to-volume ratio is from 0.5% to 10% (m/v), more preferably from 1% to 5% (m/v).
  • isotonic regulators such as glucose, mannitol, xylitol, fructose, lactose, maltose, sucrose, trehalose, glycerol, glycine, group Neither a lysine nor an arginine can be used in combination with liraglutide, and liraglutide is easily crystallized from the preparation.
  • xylitol is used instead of propylene glycol in combination with surfactants, it can also act as an isotonic regulator and a stabilizer to promote stable storage of liraglutide and not easy to crystallize. Precipitate.
  • the inventors have unexpectedly discovered that the formation of high molecular impurities such as dimers in liraglutide preparations can be reduced by the addition of xylitol, which is more inferior to the test results of the liraglutide market. Well, this unexpected technical effect has never been reported in the prior art literature.
  • the stabilizer for enhancing the stability of liraglutide is preferably polysorbate 20, polysorbate 80, poloxamer 124, poloxamer 181, poloxamer. 182, poloxamer 188, poloxamer 237, poloxamer 331, poloxamer 338, poloxamer 407 or mixtures thereof, more preferably polysorbate 20, polysorbate 80, polosa 188 or a mixture thereof.
  • Suitable pharmaceutically acceptable preservatives are included in the pharmaceutical combinations of the invention. Suitable pharmaceutically acceptable preservatives may be selected from the group consisting of phenol, o-cresol, m-cresol, p-cresol, methyl p-hydroxybenzoate, propyl p-hydroxybenzoate, 2-phenoxyethanol, p-hydroxybenzene Butyl formate, 2-phenylethanol, benzyl alcohol, chlorobutanol, chlorocresol, ethyl p-hydroxybenzoate or a mixture thereof, at a concentration of about 0.1 mg/ml to 10 mg/ml, preferably at a concentration of about 1 mg/ml. To 8 mg/ml, the most preferred concentration is from about 2 mg/ml to 6 mg/ml.
  • Preferred preservatives for the pharmaceutical composition of the invention are phenol or m-cresol, either alone or in combination.
  • a more preferred preservative is phenol at a concentration of from about 0.1 mg/ml to about 10 mg/ml, preferably at a concentration of from about 1 mg/ml to about 8 mg/ml, most preferably at a concentration of from about 2 mg/ml to about 6 mg/ml.
  • the inventors have unexpectedly found polysorbate 20, polysorbate 80, poloxamer 188 or a mixture thereof at a concentration of 0.001% to 0.5% (m/v) as a stabilizer and an isotonicity adjusting agent through a large number of comparative exploration experiments.
  • Xylitol can effectively promote the long-term stability of liraglutide preparation and prevent the precipitation of liraglutide.
  • Polysorbate and poloxamer 188 may be used singly or in combination. When used alone, the concentration of polysorbate 20, polysorbate 80 or poloxamer 188 may preferably be from about 0.004% to about 0.3%, most preferably 0.02%.
  • the stabilizer and xylitol particularly prevents the formation of liraglutide and inhibits the formation of macromolecular impurities such as dimers. use.
  • the above pharmaceutical composition contains 0.1 mg/ml to 25 mg/ml of liraglutide, a pH range of 7.5-9.0, a concentration of 5-100 mmol/L of disodium hydrogen phosphate buffer, 0.001% to 0.5%.
  • the above pharmaceutical composition contains 0.1 mg/ml to 25 mg/ml of liraglutide, a pH range of 7.5-9.0, a concentration of 5-100 mmol/L of disodium hydrogen phosphate buffer, 0.001% to 0.5%.
  • polysorbate 80 and 0.001% to about 0.5% m/v) poloxamer 188, 0.5%-10% (m/v) xylitol and 0.1 mg/ml -10 mg/ Ml of phenol or m-cresol.
  • the above pharmaceutical composition contains 1 mg/ml to 15 mg/ml of liraglutide, a pH range of 7.5-8.5, a concentration of 10-30 mmol/L of disodium hydrogen phosphate buffer, 0.004% to 0.3% ( m/v) polysorbate 80 or poloxamer 188, 1%-5% (m/v) xylitol and 1 mg/ml-8 mg/ml phenol or m-cresol.
  • the above pharmaceutical composition contains 1 mg/ml to 15 mg/ml of liraglutide, a pH range of 7.5-8.5, a concentration of 10-30 mmol/L of disodium hydrogen phosphate buffer, 0.004% to 0.3% ( m/v) polysorbate 80 and 0.004% to 0.3% (m/v) poloxamer 188, 1%-5% (m/v) xylitol and 1 mg/ml-8 mg/ml phenol Or m-cresol.
  • the above pharmaceutical composition contains 3 mg/ml to 10 mg/ml of liraglutide, a pH range of 8.0-8.5, a concentration of 10-30 mmol/L of disodium hydrogen phosphate buffer, and 0.02% (m/v).
  • the above pharmaceutical composition contains 3 mg/ml to 10 mg/ml of liraglutide, a pH range of 8.0-8.5, a concentration of 10-30 mmol/L of disodium hydrogen phosphate buffer, and 0.01% (m/v).
  • the above pharmaceutical preparation can be prepared as a lyophilized powder needle, and a pharmaceutically acceptable diluent can be added to reduce the lyophilized preparation to a state of the preparation solution before lyophilization, for example, by adding an injection. Dissolved in water. Lyophilization can be carried out using techniques commonly employed in the art, such as lyophilization cycles including freezing, primary drying, and secondary drying. Since the liquid preparation before lyophilization is substantially isotonic and/or isotonic, an appropriate amount of water for injection can be reduced to form an isotonic or isotonic solution after lyophilization.
  • the above pharmaceutical preparations can be used for the treatment of type 2 diabetes patients for improving blood sugar control in patients, and can be used alone or in combination with metformin, sulfonylureas and the like.
  • it is subcutaneous injection.
  • a preferred dosage regimen is once daily, 0.6 mg/day in the first week and 1.2 mg/day in the future. If the dose of 1.2 mg fails to control blood glucose significantly, the dose is increased to 1.8 mg. .
  • the present invention relates to a process for the preparation of a liraglutide pharmaceutical composition, wherein the preservative, buffer and stabilizer are defined as described above, and the preferred liraglutide pharmaceutical composition is as described above.
  • the solution obtained in the step (3) can be filtered by a 0.22 ⁇ m filter, and can be used for formulation potting.
  • Example 2 examines the osmotic pressure of solutions containing different osmotic pressure regulators
  • Isotonic agent Osmolality Negative control (no isotonicity) 0.041 Methionine (15mg/ml) 0.141 Glycine (15mg/ml) 0.301 Xylitol (28mg/ml) 0.284 PEG400 (61mg/ml) 0.291 L-arginine (25mg/ml) 0.322 Sorbitol (32mg/ml) 0.277 Glycerin (16.8mg/ml) 0.289 Sodium chloride (8.6mg/ml) 0.307 Imported preparation 0.281
  • the isotonic solution has an osmolality of from about 0.285 to 0.310 osmol/L.
  • Example 3 examines the stability of a formulation solution containing different stabilizers
  • the preservative, isotonic agent and buffer are dissolved in water for injection, and the crude powder of liraglutide is dissolved in the above solution while stirring slowly, and then the pH is adjusted to the desired pH with sodium hydroxide and/or hydrochloric acid. , respectively, add a certain amount of stabilizer. Finally, the above preparation solution was filtered through a 0.22 ⁇ m filter. The types and amounts of stabilizers added are shown in Table 3.
  • the composition of the formulations is as follows:
  • the low concentration of polysorbate 80 or poloxamer 188 can effectively increase the stability of the liraglutide preparation, and even under accelerated conditions, the purity of the preparation can be controlled to remain above 95%, which is stable and stable.
  • the sexual effect is stronger than that of hydroxypropyl- ⁇ -cyclodextrin, povidone K30 or PEG300, and even better than the formulation of imported products.
  • the formulation solutions of Nos. 1-4 were clarified, and no visible foreign matter or crystallization was observed.
  • the preservative, isotonic agent and buffer are dissolved in water for injection, and the crude powder of liraglutide is dissolved in the above solution while stirring slowly, and then the pH is adjusted to the desired pH with sodium hydroxide and/or hydrochloric acid. , add a certain amount of poloxamer 188. Finally, the above preparation solution was filtered through a 0.22 ⁇ m filter. The amount of 188 added to the poloxamer is shown in Table 7.
  • the composition of the preparation is as follows:
  • Formulation number The amount of poloxamer 188 1 Imitation of imported products Victoza listing prescription 2 0 3 0.004% 4 0.01% 5 0.02% 6 0.03% 7 0.04% 8 0.05% 9 0.1% 10 0.2%

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Epidemiology (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Endocrinology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Diabetes (AREA)
  • Immunology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Molecular Biology (AREA)
  • Biochemistry (AREA)
  • Zoology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Inorganic Chemistry (AREA)
  • Dermatology (AREA)
  • Hematology (AREA)
  • Emergency Medicine (AREA)
  • Obesity (AREA)
  • Organic Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Medicinal Preparation (AREA)

Abstract

一种含有GLP-1类似物的药用组合物，包括GLP-1类似物，缓冲剂，稳定剂，等渗调节剂和防腐剂。优点在于通过添加一些能被人体接受的成分，增强了GLP-1类似物的理化与生物学活性的稳定，从而制备出一种适合于临床多次注射使用的制剂。这种制剂可以有效的防止有效成分GLP-1类似物由于降解、氧化、沉淀等多种因素而导致失效或结晶，从而方便运输、长期保存和临床使用。

Description

一种包含GLP-1类似物的药物制剂及其制备方法 技术领域
本发明涉及多肽药物制剂领域，更具体地说本发明涉及一种包含GLP-1类似物的药物制剂及其制备方法。
背景技术
上世纪60年代，麦金太尔((McIntyre)和埃尔里克(Elrick)等人发现，口服葡萄糖对胰岛素分泌的促进作用明显高于静脉注射，这种额外的效应被称为“肠促胰素效应”。随着细胞和分子生物学的发展，研究证实肠促胰素是人体内一种肠源性激素，在进食后该类激素可促进胰岛素分泌，发挥葡萄糖浓度依赖性降糖作用。
肠促胰素主要由胰高血糖素-1(GLP-1)和糖依赖性胰岛素释放肽(GIP)组成，其中GLP-1在2型糖尿病的发生发展中起着更为重要的作用。研究表明，一方面GLP-1可通过刺激胰岛素分泌、抑制胰高血糖素分泌及胃排空来降低血糖；另一方面，GLP-1还有减缓β细胞凋亡，促进其再生的独特作用。
1983年Mclntyre等在分析胰高血糖素前体——胰高血糖素原(proglucagon，PG)的基因序列时发现了GLP-1。GLP-1基因在胰腺α细胞、肠道L细胞表达。完整的GLP-1多肽由37个氨基酸构成，其1-37肽序列结构为：
HDEFERHAEGTFTSDVSSYLEGQAAKEFIAWLVKGRG；
GLP-1多肽在体内的生物活性形式为GLP-1(7-37)多肽和GLP-1(7-36)酰胺，其中约80％的GLP-1循环活性来自GLP-1(7-36)酰胺。根据本领域习惯，GLP-1(7-37)OH的氨基末端指定为7号，而羧基 末端指定为37号。关于GLP-1类似物和衍生物的更详细描述见Hoffmann，J.A.[WO99/29336，1999年6月17日公布]和Knudsen，L.B.等[J.Med.Chem.43：1664-1669(2000)]。GLP-1(1-37)在体内生成后，通过两步酶切，分别去除N端6个氨基酸及形成C端酰胺化，最终生成具有高度活性的GLP-1(7-36)酰胺(也称GLP-1片段)。
GLP-1是已发现的促胰岛素分泌作用最强的肠肽类激素，它通过与GLP-1受体(GLP-1R，属于β受体家族的G偶联蛋白)结合发挥作用。GLP-1结合GLP-1R后，激活细胞膜内环腺苷酸(cAMP)和丝裂原激活蛋白激酶(MAPK)通路。胰岛成熟β细胞的GLP-1受体偶联Gs，活化腺苷酰环化酶，产生cAMP，后者与葡萄糖协同刺激胰岛素合成和分泌，刺激胰岛素基因转录和胰岛素原生物合成，降低胰高血糖素浓度并抑制胰高血糖素分泌，增强细胞对胰岛素的敏感性，刺激胰岛素依赖性糖原合成，降低餐后血糖浓度。通过激活蛋白激酶、磷脂酰肌醇3激酶(PI3K)、MAPK通道，调节凋亡前蛋白及诱导抗凋亡蛋白Bcl-2与Bcl-xL的表达以减缓β细胞凋亡，增进其再生，促使胰岛β细胞分化并增殖。此外，GLP-1还能减慢胃排空速度；通过作用下丘脑，抑制食欲。
目前GLP-1及其类似物等多肽多采用基因重组方法制备。基因重组制备蛋白、多肽常采用大肠杆菌和酵母作为宿主。其中，已上市并占据GLP-1类似物药物市场主流的药物为利拉鲁肽,商品名Victoza。利拉鲁肽(liraglutide)化学名Arg34，Lys26(Nε-(γ-Glu(Nα-十六烷酰基)))GLP-1(7-37)，是脂肪酸修饰的GLP-1多肽类似物，修饰后的药物经皮下给药后血浆半衰期延长为13小时。GLP-1多肽链序列与人GLP-1(7-37)序列类似，将人GLP-1天然序列第34位赖氨酸取代成精氨酸，其氨基酸序列与类胰高血糖素glucagon-like peptide-1(GLP-1)有约97％的同源性(Goke等，J.Biol.Chem.,268:19650-55,1993)。16碳脂肪酸通过一个谷氨酸和多肽第26位赖氨酸连接，其结构如下所示：
Figure PCTCN2016081843-appb-000001
利拉鲁肽作为GLP-1类似物，因与GLP-1高度同源而具有以下药理学作用：(1)血糖浓度依赖性促胰岛素分泌作用；(2)抑制餐后胰高血糖素分泌；(3)降低食欲，减缓胃排空。此外，利拉鲁肽可促进B细胞增殖、分化，调节B细胞凋亡基因相关表达，抑制其凋亡，增加体内胰岛B细胞数量。研究表明，利拉鲁肽亦能恢复人胰岛B细胞对血糖的敏感性。
通常治疗Ⅱ型糖尿病的一线药物是口服降血糖药，但由于目前口服降血糖药的不良反应，尤其是低血糖反应，使之并不适用于所有Ⅱ型糖尿病患者。研究表明，利拉鲁肽具有良好的糖尿病治疗前景。在全球40多个国家的5000多名糖尿病患者中进行的LEAD(Liraglutide Effect and Action in Diabetes)研究证明，利拉鲁肽具有保护胰岛B细胞功能的作用，改善B细胞胰岛素分泌的数量和质量，因此有可能从根本上改变糖尿病的发展进程。LEAD研究包括6个Ⅲ期临床试验，对利拉鲁肽单药治疗和与其他口服降糖药合用的疗效进行了广泛的评价。研究证实，利拉鲁肽与其他降糖药相比，不仅能有效保护胰岛B细胞功能，延缓Ⅱ型糖尿病的进展，而且能够迅速、高效、持久地降低糖化血红蛋白(HbA1c)，极少发生低血糖，同时还有降低体重、降低收缩压的作用。
临床试验发现利拉鲁肽对心血管系统具有一定保护作用。研究发现利拉鲁肽可以使收缩压下降2.7～6.7mmHg，此现象出现在患者体重下 降之前，因此不能把收缩压的下降完全归因于患者体重的变化。研究发现接受利拉鲁肽治疗的患者脉率略有增加，此现象的临床意义还有待进一步研究。Courreges等报道了利拉鲁肽治疗还伴随着总胆固醇、低密度脂蛋白、游离脂肪酸、甘油三酯、纤溶酶原激活物抑制因子-1及B型利钠肽浓度的下降。研究还发现利拉鲁肽治疗组患者超敏C反应蛋白水平呈现尽管不显著但为剂量依赖性的下降。
利拉鲁肽作为GLP-1类似物，其对血糖的降低作用呈葡萄糖浓度依赖性，仅在血糖浓度较高时促进胰岛素分泌并抑制胰高血糖作用，因此利拉鲁肽单独应用几乎不会引起低血糖。临床试验表明利拉鲁肽单药治疗的低血糖发生率明显低于格列美脲，与二甲双胍相当。利拉鲁肽联合治疗导致的低血糖事件与其他降糖药相当或更少。Nauck等报道利拉鲁肽联合二甲双胍治疗组患者低血糖发生率与安慰剂加二甲双胍组患者相当，均小于3％，而格列美脲联合二甲双胍治疗组患者低血糖发生率为17％，差异有统计学意义(P＜0.001)。
利拉鲁肽治疗中最常见的不良反应为胃肠道反应，主要表现为恶心、呕吐、腹泻，通常见于用药第1周，呈剂量依赖性。在LEAD-2和LEAD-3临床试验中，利拉鲁肽治疗组出现恶心的患者＜10％。通过缓慢增加剂量可减少胃肠道反应的发生，患者常在用药1～3周内耐受此不良反应，很少有患者因为胃肠道反应终止治疗。
作为新一代降血糖药，利拉鲁肽无论单独应用还是与其他口服降糖药联用，均能迅速、高效地降低血糖和HbA1c水平。其降糖作用依赖于葡萄糖浓度，即仅在血糖水平升高时刺激胰岛素释放达到治疗效果，因此发生低血糖的概率非常低。最引人瞩目的是其延缓糖尿病进展的潜力。大量临床试验表明，利拉鲁肽能够改善B细胞功能，减少其凋亡，增加B细胞的新生，从而延缓糖尿病的进展。利拉鲁肽对心血管系统的保护作用使其可以减少糖尿病心血管并发症的发生。另外，利拉鲁肽有明显的降低体重的作用，适用于需要减轻体重及有严重低血糖风险的患 者。利拉鲁肽独特的药理学作用使其在糖尿病治疗中具有广阔的前景。
利拉鲁肽的生产工艺早有报道，如参见中国专利97198413.1和99808706.8等。需要指出的是，GLP-1蛋白是一类较易降解的蛋白，有多种水解酶易促使其降解(M.Egel-Mitani，et al；Yield improvement of heterologous peptides expressed in yps1-disrupted Saccharomyces cerevisiae strains；Enzyme and Microbial Technology,2000,26:671-677)。利拉鲁肽虽然经过脂肪酸链修饰更为稳定，但是作为一种多肽类药物，其稳定性无法与常规化学药物相比，其理化、生物学性质在长期贮存时会因受到多种环境因素的影响而发生变化。例如对温度，氧和紫外线高度敏感。由于这些因素的作用，可能发生多种物理或化学变化，例如吸附，聚合，沉淀和氧化，这给其制剂处方的选择带来极大困难，在药物保质期内保证有90％以上的纯度是行业内公认的标准，比通常的蛋白药物制剂纯度需在95％以上的标准要低很多。因此如果贮存期间利拉鲁肽的稳定性不能保证的话，会导致给药剂量的变化从而影响疗效。
制剂溶液中可见异物形成的通常由下列原因引起：物理振荡和溶液中蛋白质分子的相互作用以及在贮存小瓶中液-气界面上蛋白质分子的相互作用。人们认为蛋白质分子吸附在气-液界面上，其疏水基团向空气中伸展而亲水基团浸在水相中。一旦在表面这样排布，蛋白质分子便容易聚集、形成颗粒和沉淀。人们还认为在诸如由于运输或其它过程中发生的振动而导致界面的伸缩过程制剂中，吸附于气-液和固-液界面的蛋白质发生进一步的构象变化。这种振荡能引起蛋白质缠结、聚集、形成颗粒并最终与其它吸附的蛋白质沉淀。另外，蛋白质在冻干过程中由于预冻速率、升温速率等冻干曲线工艺的影响导致可见异物产生。
CN200480034152.8号中国专利申请提及一种利拉鲁肽的改进制剂。在以往的实验中发现利拉鲁肽易从药剂溶液中结晶析出，例如包含甘露醇的制剂由于其结晶导致生产设备或者注射器的针管导致堵塞，从而影响设备的生产控制或者患者的治疗。经过大量的探索实验，在排除 甘露醇、甘油、蔗糖、PEG400、精氨酸、二甲砜、山梨糖醇、肌醇、葡萄糖、甘氨酸、麦芽糖和乳糖等常用的等渗调节剂后，发明人最终选择丙二醇作为利拉鲁肽的等渗调节剂，并且发现含有丙二醇的制剂对利拉鲁肽的物理和化学稳定性没有影响(但未提供数据证明)，制剂不易产生沉积物。
但是，由于利拉鲁肽药物是用于治疗糖尿病，需要终身给药并且每天注射。利拉鲁肽的上市处方为：3ml溶液含有18mg利拉鲁肽，1.42mg二水磷酸氢二钠，14mg丙二醇，5.5mg苯酚，溶解于注射用水，pH为8.15。虽然丙二醇是相对安全的一种要用辅料，但是根据临床总结，丙二醇对人体皮肤还是有一些可能的副作用，主要有以下几种：
(1)刺激性：有些人使用时会有主观上的灼热感、刺痛感及痒感。
(2)去脂性：丙二醇具有脂溶性溶剂的特性，长期使用高浓度丙二醇，对表皮皮脂结构会有影响。
(3)刺激性皮肤炎：丙二醇对皮肤及黏膜都具有刺激性，浓度越高、越密封的状况下使用，刺激性越大，会引起皮肤发红、起红疹、脱皮刺痒及粗糙的情形。虽然对多数人来说刺激性不大，但长期使用所可能产生的累积效应，却很值得注意。对于受伤性皮肤或敏感性肤质而言，即使是低浓度的丙二醇，也很容易产生刺激反应。
(4)过敏性皮肤炎：约有1～5％的人接触到丙二醇，会产生局部皮肤过敏性湿疹反应。通常首次接触时不会发生问题，但再度接触到时，就会产生皮肤的过敏反应。
(5)全身性接触性皮肤炎：少数对于丙二醇产生皮肤过敏反应的人，若服用或施加含丙二醇成分的食品或药品，会引起全身性皮肤过敏反应。
总体而言，丙二醇的用途相当广泛且安全性还算高，但对于皮肤仍有潜在可能的刺激性及致敏性。以其常见的用途来看，如果有其它更安全的替代成分时，能尽量减少丙二醇的使用或降低其使用浓度，对皮肤 而言会更安全。
因此，研究出一种能替代丙二醇作为利拉鲁肽制剂的药用辅料，使其不易降解，防止从制剂溶液中结晶析出，并适合于实际临床使用的药物制剂是极其有意义的。本发明涉及的就是此方面内容。
发明内容
本发明者通过大量的处方筛选和稳定性试验，意外的获得了一种稳定的包含GLP-1类似物的药用组合物。
本发明所制备的药用组合物，含有0.1mg/ml-25mg/ml的GLP-1类似物，pH7.5-9.0的缓冲剂，0.001％至0.05％(m/v)的稳定剂，0.5％-10％(m/v)的木糖醇和0.1mg/ml-10mg/ml的防腐剂。
在本申请中，名称“GLP-1类似物”意指GLP-1或其突变体，GLP-1(7-36)-酰胺或其突变体，GLP-1(7-37)或其突变体，以及上述多肽或其突变体的一个或多个氨基酸残基上引入有机取代基例如酯、烷基或亲脂性官能团化学修饰后的衍生物。突变体的含义为母体肽GLP-1，GLP-1(7-36)-酰胺或GLP-1(7-37)的一个或多个氨基酸残基已经被其他氨基酸残基取代和/或其中母体肽的一个或多个氨基酸残基已经缺失和/或其中已经将一个或多个氨基酸残基添加至母体肽。这样的添加可以发生在母体肽的N-末端或C-末端或两端。优选的，其中母体肽的6个或更少的氨基酸已经被取代和/或添加和/或缺失，更优选是这样的肽，其中母体肽的3个或更少的氨基酸已经被取代和/或添加和/或缺失，最优选是这样的肽，其中母体肽的一个氨基酸已经被取代和/或添加和/或缺失。
在本发明的一个实施方案中，GLP-1衍生物优选具有三个连接于母体肽(即，GLP-1(7-36)-酰胺，GLP-1(7-37)，GLP-1(7-36)-酰胺类似物 或GLP-1(7-37)类似物)的亲脂性取代基，更优选两个亲脂性取代基，且最优选一个亲脂性取代基，其中每个亲脂性取代基优选具有4-40个碳原子，更优选8-30个碳原子，更优选8-25个碳原子，更优选12-25个碳原子，最优选14-18个碳原子。
适合本发明的GLP-1类似物在现有技术中有多处记载，具体可参考描述于WO93/19175中(Novo Nordisk)，WO99/43705(Novo Nordisk)WO99/43706(Novo Nordisk)，WO99/43707(Novo Nordisk)，WO98/08871(具有亲脂性取代基的类似物)和WO02/46227(与血清白蛋白或与Ig的Fc部分融合的类似物)(Novo Nordisk A/S)，WO99/43708(Novo Nordisk A/S)，WO99/43341(Novo Nordisk A/S)，WO87/06941(The General Hospital Corporation)，WO90/11296(The General Hospital Corporation)，WO91/11457(Buckley等)，WO98/43658(Eli Lilly&Co.)，EP0708179-A2(Eli Lilly&Co.)，EP0699686-A2(Eli Lilly&Co.)，WO01/98331(Eli Lilly&Co.)和CN200480034152.8中所提及的那些，在此全部引入作为参考。
在本发明的一个实施方案中，GLP-1类似物优选为Arg34，Lys26(Nε-(γ-Glu(Nα-十六烷酰)))-GLP-1(7-37)，即利拉鲁肽。
在本发明的另一个实施方案中，GLP-1类似物选自：
Gly8-GLP-1(7-36)-酰胺，Gly8-GLP-1(7-37)，Val8-GLP-1(7-36)-酰胺，Val8-GLP-1(7-37)，Val8Asp22-GLP-1(7-36)-酰胺，
Val8Asp22-GLP-1(7-37)，Val8Glu22-GLP-1(7-36)-酰胺，
Val8Glu22-GLP-1(7-37)，Val8Lys22-GLP-1(7-36)-酰胺，
Val8Lys22-GLP-1(7-37)，Val8Arg22-GLP-1(7-36)-酰胺，Val8Arg22-GLP-1(7-37)，Val8His22-GLP-1(7-36)-酰胺，Val8His22-GLP-1(7-37)，及其类似物和这些中任一种的衍生物。
在本发明的另一个实施方案中，GLP-1类似物选自Arg26-GLP-1(7-37)；
Arg34-GLP-1(7-37)；Lys36-GLP-1(7-37)；Arg26，34Lys36-GLP-1(7-37)；
Arg26，34-GLP-1(7-37)；Arg26，34Lys40-GLP-1(7-37)；Arg26Lys36-GLP-1(7-37)；
Arg34Lys36-GLP-1(7-37)；Val8Arg22-GLP-1(7-37)；Met8Arg22-GLP-1(7-37)；
Gly8His22-GLP-1(7-37)；Val8His22-GLP-1(7-37)；Met8His22-GLP-1(7-37)；
His37-GLP-1(7-37)；Gly8-GLP-1(7-37)；Val8-GLP-1(7-37)；
Met8-GLP-1(7-37)；Gly8Asp22-GLP-1(7-37)；Val8Asp22-GLP-1(7-37)；
Met8Asp22-GLP-1(7-37)；Gly8Glu22-GLP-1(7-37)；Val8Glu22-GLP-1(7-37)；
Met8Glu22-GLP-1(7-37)；Gly8Lys22-GLP-1(7-37)；Val8Lys22-GLP-1(7-37)；
Met8Lys22-GLP-1(7-37)；Gly8Arg22-GLP-1(7-37)；Val8Lys22His37-GLP-1
(7-37)；Gly8Glu22His37-GLP-1(7-37)；Val8Glu22His37-GLP-1(7-37)；
Met8Glu22His37-GLP-1(7-37)；Gly8Lys22His37-GLP-1(7-37)；
Met8Lys22His37-GLP-1(7-37)；Gly8Arg22His37-GLP-1(7-37)；
Val8Arg22His37-GLP-1(7-37)；Met8Arg22His37-GLP-1(7-37)；
Gly8His22His37-GLP-1(7-37)；Val8His22His37-GLP-1(7-37)；
Met8His33His37-GLP-1(7-37)；Gly8His37-GLP-1(7-37)；
Val8His37-GLP-1(7-37)；Met8His37-GLP-1(7-37)；
Gly8Asp22His37-GLP-1(7-37)；Val8Asp22His37-GLP-1(7-37)；
Met8Asp22His37-GLP-1(7-37)；Arg26-GLP-1(7-36)-酰胺；
Arg34-GLP-1(7-36)-酰胺；Lys36-GLP-1(7-36)-酰胺；
Arg26，34Lys36-GLP-1(7-36)-酰胺；Ar26，34-GLP-1(7-36)-酰胺；
Arg26，34Lys40-GLP-1(7-36)-酰胺；Arg26Lys36-GLP-1(7-36)-酰胺；
Arg34Lys36-GLP-1(7-36)-酰胺；Gly8-GLP-1(7-36)-酰胺；
Val8-GLP-1(7-36)-酰胺；Met8-GLP-1(7-36)-酰胺；
Gly8Asp22-GLP-1(7-36)-酰胺；Gly8Glu22His37-GLP-1(7-36)-酰胺；
Val8Asp22-GLP-1(7-36)-酰胺；Met8Asp22-GLP-1(7-36)-酰胺；
Gly8Glu22-GLP-1(7-36)-酰胺；Val8Glu22-GLP-1(7-36)-酰胺；
Met8Glu22-GLP-1(7-36)-酰胺；Gly8Lys22-GLP-1(7-36)-酰胺；
Val8Lys22-GLP-1(7-36)-酰胺；Met8Lys22-GLP-1(7-36)-酰胺；
Gly8His22His37-GLP-1(7-36)-酰胺；Gly8Arg22-GLP-1(7-36)-酰胺；
Val8Arg22-GLP-1(7-36)-酰胺；Met8Arg22-GLP-1(7-36)-酰胺；
Gly8His22-GLP-1(7-36)-酰胺；Val8His22-GLP-1(7-36)-酰胺；
Met8His22-GLP-1(7-36)-酰胺；His37-GLP-1(7-36)-酰胺；
Val8Arg22His37-GLP-1(7-36)-酰胺；Met8Arg22His37-GLP-1(7-36)-酰胺；
Gly8His37-GLP-1(7-36)-酰胺；Val8His37-GLP-1(7-36)-酰胺；
Met8His37-GLP-1(7-36)-酰胺；Gly8Asp22His37-GLP-1(7-36)-酰胺；
Val8Asp22His37-GLP-1(7-36)-酰胺；Met8Asp22His37-GLP-1(7-36)-酰胺；
Val8Glu22His37-GLP-1(7-36)-酰胺；Met8Glu22His37-GLP-1(7-36)-酰胺；
Gly8Lys22His37-GLP-1(7-36)-酰胺；Val8Lys22His37-GLP-1(7-36)-酰胺；
Met8Lys22His37-GLP-1(7-36)-酰胺；Gly8Arg22His37-GLP-1(7-36)-酰胺；
Val8His22His37-GLP-1(7-36)-酰胺；Met8His22His37-GLP-1(7-36)-酰胺；
及其衍生物。
在本发明的另一个实施方案中，GLP-1类似物选自
Val8Trp19Glu22-GLP-1(7-37)，Val8Glu22Val25-GLP-1(7-37)，
Val8Tyr16Glu22-GLP-1(7-37)，Val8Trp16Glu22-GLP-1(7-37)，
Val8Leu16Glu22-GLP-1(7-37)，Val8Tyr18Glu22-GLP-1(7-37)，
Val8Glu22His37GLP-1(7-37)，Val8Glu22Ile33-GLP-1(7-37)，
Val8Trp16Glu22Val25Ile33-GLP-1(7-37)，Val8Trp16Glu22Ile33-GLP-1(7-37)，Val8Glu22Val25Ile33-GLP-1(7-37)，Val8Trp16Glu22Val25-GLP-1(7-37)，
及其类似物和这些中任一种的衍生物。
本发明公开的药物组合物中，GLP-1类似物的浓度优选为约1mg/ml-15mg/ml，更优选为3mg/ml-10mg/ml，最优选为6mg/ml。
本发明公开的药物组合物中，适用于本发明的缓冲剂为能够维持制剂在水溶液状态下pH值为7.5-9.0的任何一种缓冲液，可任选自磷酸 缓冲液、磷酸氢二钠-柠檬酸缓冲液、TRIS、甘氨酰基-甘氨酸、N-二(羟乙基)甘氨酸、磷酸二氢钠缓冲液、磷酸氢二钠缓冲液、醋酸钠缓冲液、碳酸钠缓冲液、磷酸钠缓冲液、赖氨酸缓冲液、精氨酸缓冲液或其混合物。缓冲剂的pH值范围优选为7.5-8.5，更优选为8.0-8.5；缓冲剂的浓度为5-100mmol/L，优选10-30mmol/L。优选磷酸氢二钠缓冲液,浓度为5-100mmol/L，pH范围在7.5-8.5之间；更优选磷酸氢二钠缓冲液浓度为10-30mmol/L,pH范围为8.0-8.5。
本发明公开的药物组合物中，使用木糖醇作为等渗调节剂，重量体积比浓度为0.5％-10％(m/v)，更优选为1％-5％(m/v)。
本发明公开的药用组合物中，用于提高GLP-1类似物稳定性的稳定剂包括但不限于：氨基酸及衍生物：甘氨酸、丙氨酸、丝氨酸、天冬氨酸、谷氨酸、苏氨酸、色氨酸、赖氨酸、羟赖氨酸、组氨酸、精氨酸、胱氨酸、半胱氨酸、甲硫氨酸、苯丙氨酸、亮氨酸、异亮氨酸等以及它们的衍生物；非离子表面活性剂：脱水山梨醇脂肪酸酯，甘油脂肪酸酯(如脱水山梨糖醇辛酸单酯，脱水山梨糖醇月桂酸单酯和脱水山梨糖醇棕榈酸单酯)，聚甘油脂肪酸酯(例如甘油辛酸单酯，甘油肉豆蔻酸单脂霜和甘油硬脂肪酸单酯)，聚氧乙烯脱水山梨醇脂肪酸酯，聚氧乙烯山梨醇脂肪酸酯，聚氧乙烯甘油脂肪酸酯，聚氧乙烯乙二醇脂肪酸酯，聚氧乙烯烷基醚，聚氧乙烯聚氧丙烯烷基醚，聚氧乙烯苯醚，聚氧乙基化硬蓖麻油，聚氧乙基化蜂蜡衍生物，聚氧乙烯化羊毛脂衍生物或者聚氧乙烯脂肪酸酰胺，阳离子表面活性剂是烷基硫酸盐(如有—个C10—C18烷基的烷基硫酸盐)；聚乙二醇，聚乙烯醇，羟丙基－β－环糊精，羧甲基纤维素，聚乙烯吡咯烷酮,聚山梨酯20或聚山梨酯80，泊洛沙姆系列(泊洛沙姆124、泊洛沙姆181、泊洛沙姆182、泊洛沙姆188、泊洛沙姆237、泊洛沙姆331、泊洛沙姆338或泊洛沙姆407)。所述的稳定剂优选聚山梨酯20、聚山梨酯80、泊洛沙姆124、泊洛沙姆181、泊洛沙姆182、泊洛沙姆188、泊洛沙姆237、泊洛沙姆331、泊洛沙姆338、 泊洛沙姆407或其混合物，更优选聚山梨酯20、聚山梨酯80、泊洛沙姆188或其混合物。
本发明公开的药物组合中包含了药学上可接受的防腐剂。合适的药学上可接受的防腐剂可以选自苯酚、邻甲酚、间甲酚、对甲酚、对羟基苯甲酸甲酯、对羟基苯甲酸丙酯、2－苯氧基乙醇、对羟基苯甲酸丁酯、2－苯基乙醇、苯甲醇、氯丁醇、氯甲酚、对羟基苯甲酸乙酯或其混合物，浓度约为0.1mg/ml到10mg/ml，优选浓度约为1mg/ml到8mg/ml，最优选浓度约为2mg/ml到6mg/ml。
本发明还公开了一种GLP-1类似物药物组合物的制备方法，所述方法包括如下步骤：
(1)将防腐剂、木糖醇和缓冲剂溶解于注射用水中制得溶液；
(2)将GLP-1类似物原料溶解于上述溶液中，调节至所需的pH范围；
(3)在上述溶液中加入稳定剂，得到含有0.1mg/ml-25mg/ml的GLP-1类似物、pH7.5-9.0的缓冲剂、0.001％至0.5％(m/v)的稳定剂、0.5％-10％(m/v)的木糖醇和0.1mg/ml-10mg/ml防腐剂的GLP-1类似物药物组合物。
其中防腐剂，缓冲剂，稳定剂和GLP-1类似物的定义如前所述。
发明人经过大量实验，意外发现，本发明公开的药物组合物成分特别适用于利拉鲁肽药物制剂的长期保存。利拉鲁肽为皮下注射给药，为使利拉鲁肽制剂能稳定保存，使其不易降解和从制剂溶液中结晶析出，发明人发现需要在制剂溶液中添加适宜量的表面活性剂。但是，出于对临床应用安全性的考虑，使能混入用于注射入人体的非肠道组合物的表面活性剂的类型与特性受到限制。因此，本领域需要能提供改进的蛋白质稳定性的药物组合物，该组合物中只包含那些被认为是安全的并包括在权威批准的规章中的商用非肠道药成分。
本发明通过大量的摸索实验意外发现，利用木糖醇作为等渗调节剂替代丙二醇并配合低浓度的poloxamer表面活性剂与聚山梨酯表面活性 剂(优选吐温-20或吐温-80)单独或者联合应用可提高利拉鲁肽制剂的长期稳定性，在4℃条件下存放3个月，利拉鲁肽纯度依然保持在95％以上，比进口品Victoza上市处方更为稳定；而且制剂溶液保持澄清，未见有可见异物或者结晶导致注射剂针管堵塞等现象；而且，木糖醇在糖尿病方面的临床应用其安全性要比丙二醇高的多，副作用更少。
更为重要的是，发明人意外发现，木糖醇配合低浓度的poloxamer表面活性剂与聚山梨酯表面活性剂可以降低利拉鲁肽制剂长期保存时高分子杂质例如二聚体的生成，这一意外技术效果是现有公开技术文献中所从未报道的。
木糖醇的血糖指数(glycemic index)极低，是胰岛素的天然稳定剂，而且其代谢不依赖于胰岛素，也不会引起血糖值升高，并可消除糖尿病人三多(多饮、多尿、多食)，因此是糖尿病人安全的甜味剂、营养补充剂和辅助治疗剂。除此之外，木糖醇具有高度的化学稳定性，这意味着它不会与药物和其他辅料发生相互作用，并且能够在较宽的pH范围内(pH1-11)使用。其次，木糖醇在水中具有较低的活度和较高的渗透压，因此可增加产品的稳定性。另外，文献报道木糖醇可对防腐剂起增效作用，同时自身也具有抑菌和杀菌的性质，进而可以增强产品的稳定性。
本发明还公开了一种利拉鲁肽药物组合物的制备方法，所述方法包括如下步骤：
(1)将防腐剂、木糖醇和缓冲剂溶解于注射用水中制得溶液；
(2)将利拉鲁肽原料溶解于上述溶液中，将pH调节至所需的pH；
(3)在上述溶液中加入稳定剂，得到含有0.1mg/ml-25mg/ml的利拉鲁肽、pH7.5-9.0的缓冲剂、0.001％至0.5％(m/v)的稳定剂、0.5％-10％(m/v)的木糖醇和0.1mg/ml-10mg/ml防腐剂的利拉鲁肽药物组合物。
本发明所制备的药用组合物，含有0.1mg/ml-25mg/ml的利拉鲁肽，pH7.5-9.0的缓冲剂，0.001％至0.05％(m/v)的稳定剂，0.5％-10％(m/v) 的木糖醇和0.1mg/ml-10mg/ml的防腐剂。本发明的优点在于通过添加一些能被人体接受的成分，增强了利拉鲁肽的理化与生物学活性的稳定，从而制备出一种适合于临床多次注射使用的制剂。这种制剂可以有效的防止有效成分利拉鲁肽由于降解、氧化、沉淀等多种因素而导致的形成高分子聚合物、纯度降低或结晶析出，从而利于方便运输、长期保存和临床使用。
上述的药物组合物中，利拉鲁肽的浓度优选为约1mg/ml-15mg/ml，更优选为3mg/ml-10mg/ml，最优选为6mg/ml。
上述的药物组合物中，适用于本发明的缓冲剂为能够维持制剂在水溶液状态下pH值为7.5-9.0的任何一种缓冲液，可任选自磷酸缓冲液、磷酸氢二钠-柠檬酸缓冲液、TRIS、甘氨酰基-甘氨酸、N-二(羟乙基)甘氨酸、磷酸二氢钠缓冲液、磷酸氢二钠缓冲液、醋酸钠缓冲液、碳酸钠缓冲液、磷酸钠缓冲液、赖氨酸缓冲液、精氨酸缓冲液或其混合物。缓冲剂的pH值范围优选为7.5-8.5，更优选为8.0-8.5；缓冲剂的浓度为5-100mmol/L，优选10-30mmol/L。优选磷酸氢二钠缓冲液,浓度为5-100mmol/L，pH范围在7.5-8.5之间；更优选磷酸氢二钠缓冲液浓度为10-30mmol/L,pH范围为8.0-8.5。
本发明的药物组合中使用木糖醇作为等渗调节剂，重量体积比浓度为0.5％-10％(m/v)，更优选为1％-5％(m/v)。根据诺和诺德公司申请的CN200480034152.8号专利公开的技术内容，常用的等渗调节剂如葡萄糖，甘露醇，木糖醇、果糖，乳糖，麦芽糖，蔗糖，海藻糖，甘油，甘氨酸，组氨酸或精氨酸等均不能配合利拉鲁肽使用，易使利拉鲁肽从制剂中结晶析出。发明人经过大量的摸索实验，意外发现，若使用木糖醇替代丙二醇配合表面活性剂联合使用，可以兼起等渗调节剂和稳定剂的作用，促使利拉鲁肽制剂能够稳定保存，不易结晶析出。另外，发明人意外地发现，由于木糖醇的加入，可以降低利拉鲁肽制剂中高分子杂质例如二聚体的生成，该技术指标要比利拉鲁肽上市处方的检测结果更 好，这一意外技术效果是现有公开技术文献中所从未报道的。
本发明公开的药用组合物中，用于提高利拉鲁肽稳定性的稳定剂优选为聚山梨酯20、聚山梨酯80、泊洛沙姆124、泊洛沙姆181、泊洛沙姆182、泊洛沙姆188、泊洛沙姆237、泊洛沙姆331、泊洛沙姆338、泊洛沙姆407或其混合物，更优选聚山梨酯20、聚山梨酯80、泊洛沙姆188或其混合物。
本发明的药物组合中包含了药学上可接受的防腐剂。合适的药学上可接受的防腐剂可以选自苯酚、邻甲酚、间甲酚、对甲酚、对羟基苯甲酸甲酯、对羟基苯甲酸丙酯、2－苯氧基乙醇、对羟基苯甲酸丁酯、2－苯基乙醇、苯甲醇、氯丁醇、氯甲酚、对羟基苯甲酸乙酯或其混合物，浓度约为0.1mg/ml到10mg/ml，优选浓度约为1mg/ml到8mg/ml，最优选浓度约为2mg/ml到6mg/ml。
发明的药用组合物优选的防腐剂是苯酚或间甲酚，二者可以单独使用，也可以联合使用。更优选的防腐剂为苯酚，浓度约为0.1mg/ml到10mg/ml，优选浓度约为1mg/ml到8mg/ml，最优选浓度约为2mg/ml到6mg/ml。
发明人通过大量的对比摸索实验，意外发现浓度为0.001％至0.5％(m/v)的聚山梨酯20、聚山梨酯80、泊洛沙姆188或其混合物作为稳定剂配合等渗调节剂木糖醇可有效促进利拉鲁肽制剂的长期稳定性，防止利拉鲁肽结晶析出。聚山梨酯和泊洛沙姆188可以单独使用，也可以联合使用。单独应用时，聚山梨酯20、聚山梨酯80或泊洛沙姆188浓度可优选为约为0.004％至约0.3％，最优选为0.02％。联合应用时，含有约0.001％至约0.3％(m/v)的聚山梨酯80和0.001％至约0.3％(m/v)的泊洛沙姆188，更优选约0.004％至约0.2％(m/v)的聚山梨酯80和0.004％至约0.2％(m/v)的泊洛沙姆188，更优选为0.01％(m/v)的聚山梨酯80和0.01％(m/v)的泊洛沙姆188。该稳定剂和木糖醇的联合作用对防止利拉鲁肽的结晶形成以及抑制高分子杂质例如二聚体的生成特别有 用。
优选的，上述药物组合物中含有0.1mg/ml-25mg/ml的利拉鲁肽，pH范围为7.5-9.0、浓度为5-100mmol/L的磷酸氢二钠缓冲剂，0.001％至0.5％(m/v)的聚山梨酯80或泊洛沙姆188，0.5％-10％(m/v)的木糖醇和0.1mg/ml-10mg/ml的苯酚或间甲酚。
优选的，上述药物组合物中含有0.1mg/ml-25mg/ml的利拉鲁肽，pH范围为7.5-9.0、浓度为5-100mmol/L的磷酸氢二钠缓冲剂，0.001％至0.5％(m/v)的聚山梨酯80和0.001％至约0.5％(m/v)的泊洛沙姆188，0.5％-10％(m/v)的木糖醇和0.1mg/ml-10mg/ml的苯酚或间甲酚。
优选的，上述药物组合物中含有1mg/ml-15mg/ml的利拉鲁肽，pH范围为7.5-8.5、浓度为10-30mmol/L的磷酸氢二钠缓冲剂，0.004％至0.3％(m/v)的聚山梨酯80或泊洛沙姆188，1％-5％(m/v)的木糖醇和1mg/ml-8mg/ml的苯酚或间甲酚。
优选的，上述药物组合物中含有1mg/ml-15mg/ml的利拉鲁肽，pH范围为7.5-8.5、浓度为10-30mmol/L的磷酸氢二钠缓冲剂，0.004％至0.3％(m/v)的聚山梨酯80和0.004％至0.3％(m/v)的泊洛沙姆188，1％-5％(m/v)的木糖醇和1mg/ml-8mg/ml的苯酚或间甲酚。
优选的，上述药物组合物中含有3mg/ml-10mg/ml的利拉鲁肽，pH范围为8.0-8.5、浓度为10-30mmol/L的磷酸氢二钠缓冲剂，0.02％(m/v)的聚山梨酯80或泊洛沙姆188，1％-5％(m/v)的木糖醇和2mg/ml-6mg/ml的苯酚或间甲酚。
优选的，上述药物组合物中含有3mg/ml-10mg/ml的利拉鲁肽，pH范围为8.0-8.5、浓度为10-30mmol/L的磷酸氢二钠缓冲剂，0.01％(m/v)的聚山梨酯80和0.01％的泊洛沙姆188，1％-5％(m/v)的木糖醇和2mg/ml-6mg/ml的苯酚或间甲酚。
根据需要，可将上述的药物制剂制备为冻干粉针，加入药学上可接受的稀释剂可将冻干制剂还原成冻干前的制剂溶液状态，例如加入注射 用水溶解。可以用本领域常用技术进行冻干，例如冻干循环包括冷冻、初次干燥和二次干燥。由于冻干前的液体制剂基本上为等张和/或等渗的，因此冻干后加入适量的注射用水能够还原形成等张或等渗溶液。
上述的药物制剂可用于治疗II型糖尿病患者，用于改善患者的血糖控制，可单独使用或与二甲双胍、磺脲类药物等联合应用。临床使用时为皮下注射，一种优选的给药方案为每天给药一次，第一周0.6mg/天，以后为1.2mg/天；如果1.2mg剂量未能明显控制血糖，剂量增加至1.8mg。
本发明涉及的利拉鲁肽药物组合物的制备方法，所述方法中的防腐剂、缓冲剂和稳定剂的定义如前所述，优选的利拉鲁肽药物组合物亦如前所述。
在配制时，步骤(3)所得溶液可用0.22μm滤器过滤，即可用于制剂灌封。
为了进—步阐明本发明，提供了下列例子，这些例子仅仅是为了进一步说明本发明，并不意味着作为一种限制。
具体实施方式
实施例一考察利拉鲁肽原粉在不同pH值条件下的溶解情况
将适量的利拉鲁肽原粉分别溶解在注射用水和不同的pH的磷酸氢二钠缓冲液中，其溶解情况见表1。
表1原粉在不同的pH的溶液中的溶解情况
  利拉鲁肽的浓度 溶解情况
注射用水 6mg/ml 不溶解
10mM磷酸氢二钠缓冲液(pH7.00) 6mg/ml 不溶解
10mM磷酸氢二钠缓冲液(pH7.50) 6mg/ml 无色澄清
10mM磷酸氢二钠缓冲液(pH8.00) 6mg/ml 无色澄清
10mM磷酸氢二钠缓冲液(pH8.15) 6mg/ml 无色澄清
10mM磷酸氢二钠缓冲液(pH8.50) 6mg/ml 无色澄清
10mM磷酸氢二钠缓冲液(pH9.00) 6mg/ml 无色澄清
从上述试验可知：利拉鲁肽原粉在酸性及中性条件下不溶解，在偏碱性的条件下才溶解。
实施例二考察含有不同渗透压调节剂溶液的渗透压
将不同的渗透压调节剂溶于10mM的磷酸氢二钠缓冲液中，边搅拌边加入利拉鲁肽原粉(6mg/ml)，然后用氢氧化钠将pH值调节到pH8.15。最后，将上述溶液分别用0.22μm滤器过滤。每种溶液等渗剂的浓度和渗透压的测试结果见表2：
表2等渗剂的浓度和渗透压的测试结果
等渗剂 摩尔渗透压浓度
阴性对照(无等渗剂) 0.041
甲硫氨酸(15mg/ml) 0.141
甘氨酸(15mg/ml) 0.301
木糖醇(28mg/ml) 0.284
PEG400(61mg/ml) 0.291
L-精氨酸(25mg/ml) 0.322
山梨醇(32mg/ml) 0.277
甘油(16.8mg/ml) 0.289
氯化钠(8.6mg/ml) 0.307
进口制剂 0.281
等渗溶液具有约0.285～0.310osmol/L的摩尔渗透压浓度。
实施例三考察含有不同稳定剂的制剂溶液的稳定性
将防腐剂、等渗剂和缓冲剂溶解于注射用水中，边缓慢搅拌边将利拉鲁肽原粉溶解与上述溶液中，然后用氢氧化钠和/或盐酸将pH调节至所需的pH，分别加入一定量的稳定剂。最后将上述制剂溶液用0.22μm滤器过滤。加入稳定剂的种类和用量见表3，制剂的组成如下：
Figure PCTCN2016081843-appb-000002
表3等渗剂、稳定剂的种类和用量
Figure PCTCN2016081843-appb-000003
将上述制剂分别放入37℃、25℃和4℃进行稳定性考察,用HPLC法检测样品的有关物质(面积归一法)。具体结果如下：
表4 37℃条件下稳定性考察结果
Figure PCTCN2016081843-appb-000004
表5 25℃条件下稳定性考察结果
Figure PCTCN2016081843-appb-000005
表6 4℃条件下稳定性考察结果
Figure PCTCN2016081843-appb-000006
从以上结果可以看出，低浓度的聚山梨酯80或泊洛沙姆188能有效增加利拉鲁肽制剂的稳定性，甚至在加速条件下能控制制剂纯度保持在95％以上，其增加稳定性效果要比羟丙基-β-环糊精、聚维酮K30或PEG300等强，甚至比进口品上市处方的稳定性更好。稳定性考察期间编号1-4组制剂溶液澄清，未见有可见异物或者结晶现象。
实施例四考察不同浓度泊洛沙姆188作为稳定剂的制剂溶液稳定性
将防腐剂、等渗剂和缓冲剂溶解于注射用水中，边缓慢搅拌边将利拉鲁肽原粉溶解与上述溶液中，然后用氢氧化钠和/或盐酸将pH调节至所需的pH，分别加入一定量的泊洛沙姆188。最后将上述制剂溶液用0.22μm滤器过滤。加入泊洛沙姆188量见表7，制剂的组成如下：
Figure PCTCN2016081843-appb-000007
表7不同处方泊洛沙姆188的浓度
制剂编号 泊洛沙姆188的用量
1 仿进口品Victoza上市处方
2 0
3 0.004％
4 0.01％
5 0.02％
6 0.03％
7 0.04％
8 0.05％
9 0.1％
10 0.2％
将上述制剂分别放入37℃、25℃和4℃进行稳定性考察,用RP-HPLC法检测样品的有关物质如降解产物(面积归一法),用SEC-HPLC法检测样品高分子杂质如多聚体或/和二聚体(面积归一法)。稳定性考察期间各编号组制剂溶液澄清，未见有可见异物或者结晶现象。具体结果如下：
表8 37℃条件下稳定性考察样品有关物质结果
Figure PCTCN2016081843-appb-000008
Figure PCTCN2016081843-appb-000009
表9 25℃条件下稳定性考察样品有关物质结果
Figure PCTCN2016081843-appb-000010
表10 4℃条件下稳定性考察样品有关物质结果
Figure PCTCN2016081843-appb-000011
表11 37℃条件下稳定性考察样品高分子杂质结果
Figure PCTCN2016081843-appb-000012
表12 25℃条件下稳定性考察样品高分子杂质结果
Figure PCTCN2016081843-appb-000013
表13 4℃条件下稳定性考察样品高分子杂质结果
Figure PCTCN2016081843-appb-000014
从以上结果可以看出，添加有0.004％-0.03％泊洛沙姆188和等渗调节剂木糖醇的利拉鲁肽制剂物理稳定性和化学稳定性大为提高。在4℃条件下存放12个月，利拉鲁肽纯度依然保持在97％以上，而且制剂溶液保持澄清，未见有可见异物或者结晶导致注射剂针管堵塞等现象，尤其4℃、25℃和37℃条件下稳定性考察样品高分子杂质结果数据优于进口品上市处方。

Claims (82)

  1. 一种药物组合物，含有0.1mg/ml-25mg/ml的利拉鲁肽，pH7.5-9.0的缓冲剂，0.001％至0.5％(m/v)的稳定剂，0.5％-10％(m/v)的木糖醇和0.1mg/ml-10mg/ml的防腐剂。
  2. 根据权利要求1所述的药物组合物，其特征在于：所述组合物中利拉鲁肽的浓度1mg/ml-15mg/ml。
  3. 根据权利要求2所述的药物组合物，其特征在于：所述组合物中利拉鲁肽的浓度为3mg/ml-10mg/ml。
  4. 根据权利要求3所述的药物组合物，其特征在于：所述组合物中利拉鲁肽的浓度为6mg/ml。
  5. 根据权利要求1所述的药物组合物，其特征在于：所述组合物中缓冲剂为能够维持制剂在水溶液状态下pH值为7.5-9.0的任何一种缓冲液，可任选自磷酸缓冲液、磷酸氢二钠-柠檬酸缓冲液、TRIS、甘氨酰基-甘氨酸、N-二(羟乙基)甘氨酸、磷酸二氢钠缓冲液、磷酸氢二钠缓冲液、醋酸钠缓冲液、碳酸钠缓冲液、磷酸钠缓冲液、赖氨酸缓冲液、精氨酸缓冲液或其混合物。
  6. 根据权利要求5所述的药物组合物，其特征在于：所述组合物中缓冲剂的pH值范围为7.5至8.5。
  7. 根据权利要求6所述的药物组合物，其特征在于：所述组合物中缓冲剂的pH值范围为8.0-8.5。
  8. 根据权利要求5所述的药物组合物，其特征在于：所述组合物中缓冲剂的浓度为5-100mmol/L。
  9. 根据权利要求8所述的药物组合物，其特征在于：所述组合物中缓冲剂的浓度为10-30mmol/L。
  10. 根据权利要求5所述的药物组合物，其特征在于：所述组合物中缓冲剂为磷酸氢二钠缓冲液,浓度为5-100mmol/L，pH范围在7.5-8.5之 间。
  11. 根据权利要求10所述的药物组合物，其特征在于：所述组合物中缓冲剂为磷酸氢二钠缓冲液,浓度为10-30mmol/L,pH范围为8.0-8.5。
  12. 根据权利要求1所述的药物组合物，其特征在于：所述组合物中木糖醇的浓度为1％-5％(m/v)。
  13. 根据权利要求1所述的药物组合物，其特征在于：所述组合物中稳定剂选自聚山梨酯20、聚山梨酯80、泊洛沙姆124、泊洛沙姆181、泊洛沙姆182、泊洛沙姆188、泊洛沙姆237、泊洛沙姆331、泊洛沙姆338或泊洛沙姆407。
  14. 根据权利要求13所述的药物组合物，其特征在于：所述组合物中稳定剂选自聚山梨酯20、聚山梨酯80或泊洛沙姆188，浓度为0.004％至0.3％(m/v)。
  15. 根据权利要求14所述的药物组合物，其特征在于：所述组合物中稳定剂选自聚山梨酯20、聚山梨酯80或泊洛沙姆188，浓度为0.02％(m/v)。
  16. 根据权利要求1所述的药物组合物，其特征在于：所述组合物中稳定剂为聚山梨酯80和泊洛沙姆188的混合物，含有0.001％至0.3％(m/v)的聚山梨酯80和0.001％至0.3％(m/v)的泊洛沙姆188。
  17. 根据权利要求16所述的药物组合物，其特征在于：所述组合物中稳定剂为聚山梨酯80和泊洛沙姆188的混合物，含有0.004％至0.2％(m/v)的聚山梨酯80和0.004％至0.2％(m/v)的泊洛沙姆188。
  18. 根据权利要求16所述的药物组合物，其特征在于：所述组合物中稳定剂为聚山梨酯80和泊洛沙姆188的混合物，含有0.01％(m/v)的聚山梨酯80和0.01％(m/v)的泊洛沙姆188。
  19. 根据权利要求1所述的药物组合物，其特征在于：所述组合物中防腐剂选自苯酚、邻甲酚、间甲酚、对甲酚、对羟基苯甲酸甲酯、对羟基苯甲酸丙酯、2－苯氧基乙醇、对羟基苯甲酸丁酯、2－苯基乙醇、苯 甲醇、氯丁醇、氯甲酚、对羟基苯甲酸乙酯或其混合物，浓度为0.1mg/ml到10mg/ml。
  20. 根据权利要求19所述的药物组合物，其特征在于：所述组合物中防腐剂的浓度为1mg/ml到8mg/ml。
  21. 根据权利要求20所述的药物组合物，其特征在于：所述组合物中防腐剂的浓度为2mg/ml到6mg/ml。
  22. 根据权利要求19-21任一项所述的药物组合物，其特征在于：所述组合物中防腐剂为苯酚或间甲酚，二者可以单独使用，也可以联合使用。
  23. 根据权利要求19所述的药物组合物，其特征在于：所述组合物中防腐剂为苯酚，浓度为0.1mg/ml到10mg/ml。
  24. 根据权利要求23所述的药物组合物，其特征在于：所述组合物中防腐剂为苯酚，浓度为1mg/ml到8mg/ml。
  25. 根据权利要求24所述的药物组合物，其特征在于：所述组合物中防腐剂为苯酚，浓度为2mg/ml到6mg/ml。
  26. 一种药物组合物，含有0.1mg/ml-25mg/ml的利拉鲁肽，pH范围为7.5-9.0、浓度为5-100mmol/L的磷酸氢二钠缓冲剂，0.001％至0.5％(m/v)的聚山梨酯80或泊洛沙姆188，0.5％-10％(m/v)的木糖醇和0.1mg/ml-10mg/ml的苯酚或间甲酚。
  27. 一种药物组合物，含有0.1mg/ml-25mg/ml的利拉鲁肽，pH范围为7.5-9.0、浓度为5-100mmol/L的磷酸氢二钠缓冲剂，0.001％至0.5％(m/v)的聚山梨酯80和0.001％至约0.5％(m/v)的泊洛沙姆188，0.5％-10％(m/v)的木糖醇和0.1mg/ml-10mg/ml的苯酚或间甲酚。
  28. 一种药物组合物，含有1mg/ml-15mg/ml的利拉鲁肽，pH范围为7.5-8.5、浓度为10-30mmol/L的磷酸氢二钠缓冲剂，0.004％至0.3％(m/v)的聚山梨酯80或泊洛沙姆188，1％-5％(m/v)的木糖醇和1mg/ml-8mg/ml的苯酚或间甲酚。
  29. 一种药物组合物，含有1mg/ml-15mg/ml的利拉鲁肽，pH范围为 7.5-8.5、浓度为10-30mmol/L的磷酸氢二钠缓冲剂，0.004％至0.3％(m/v)的聚山梨酯80和0.004％至0.3％(m/v)的泊洛沙姆188，1％-5％(m/v)的木糖醇和1mg/ml-8mg/ml的苯酚或间甲酚。
  30. 一种药物组合物，含有3mg/ml-10mg/ml的利拉鲁肽，pH范围为8.0-8.5、浓度为10-30mmol/L的磷酸氢二钠缓冲剂，0.02％(m/v)的聚山梨酯80或泊洛沙姆188，1％-5％(m/v)的木糖醇和2mg/ml-6mg/ml的苯酚或间甲酚。
  31. 一种药物组合物，含有3mg/ml-10mg/ml的利拉鲁肽，pH范围为8.0-8.5、浓度为10-30mmol/L的磷酸氢二钠缓冲剂，0.01％(m/v)的聚山梨酯80和0.01％的泊洛沙姆188，1％-5％(m/v)的木糖醇和2mg/ml-6mg/ml的苯酚或间甲酚。
  32. 根据权利要求1-31任一项所述药物组合物，其特征在于：所述药物组合物可制备为冻干粉针。
  33. 利拉鲁肽在制备权利要求1-31任一项所述的药物组合物中的用途，所述的药物组合物用于治疗II型糖尿病。
  34. 根据权利要求33所述的用途，其特征在于：所述的药物组合物可单独使用或与二甲双胍、磺脲类药物等联合应用。
  35. 根据权利要求34所述的用途，其特征在于：所述的药物组合物
    临床使用时为皮下注射。
  36. 根据权利要求35所述的用途，其特征在于：所述的药物组合物给药方案为每天给药一次，第一周0.6mg/天，以后为1.2mg/天；如果1.2mg剂量未能明显控制血糖，剂量增加至1.8mg。
  37. 一种利拉鲁肽药物组合物的制备方法，所述方法包括如下步骤：
    (1)将防腐剂、木糖醇和缓冲剂溶解于注射用水中制得溶液；
    (2)将利拉鲁肽原料溶解于上述溶液中，将pH调节至所需的pH；
    (3)在上述溶液中加入稳定剂，得到如权利要求1所述的药物组合物。
  38. 根据权利要求37所述的制备方法，其特征在于：所述方法制备得 到的组合物中利拉鲁肽的浓度1mg/ml-15mg/ml。
  39. 根据权利要求38所述的制备方法，其特征在于：所述方法制备得到的组合物中利拉鲁肽的浓度为3mg/ml-10mg/ml。
  40. 根据权利要求39所述的制备方法，其特征在于：所述方法制备得到的组合物中利拉鲁肽的浓度为6mg/ml。
  41. 根据权利要求37所述的制备方法，其特征在于：所述方法步骤(1)中缓冲剂为能够维持制剂在水溶液状态下pH值为7.5-9.0的任何一种缓冲液，可任选自磷酸缓冲液、磷酸氢二钠-柠檬酸缓冲液、TRIS、甘氨酰基-甘氨酸、N-二(羟乙基)甘氨酸、磷酸二氢钠缓冲液、磷酸氢二钠缓冲液、醋酸钠缓冲液、碳酸钠缓冲液、磷酸钠缓冲液、赖氨酸缓冲液、精氨酸缓冲液或其混合物。
  42. 根据权利要求41所述的制备方法，其特征在于：所述方法步骤(1)中缓冲剂的pH值范围为7.5至8.5。
  43. 根据权利要求42所述的制备方法，其特征在于：所述方法步骤(1)中缓冲剂的pH值范围为8.0-8.5。
  44. 根据权利要求41所述的制备方法，其特征在于：所述方法步骤(1)中缓冲剂的浓度为5-100mmol/L。
  45. 根据权利要求44所述的制备方法，其特征在于：所述方法步骤(1)中缓冲剂的浓度为10-30mmol/L。
  46. 根据权利要求41所述的制备方法，其特征在于：所述方法步骤(1)中缓冲剂为磷酸氢二钠缓冲液,浓度为5-100mmol/L，pH范围在7.5-8.5之间。
  47. 根据权利要求46所述的制备方法，其特征在于：所述方法步骤(1)中缓冲剂为磷酸氢二钠缓冲液,浓度为10-30mmol/L,pH范围为8.0-8.5。
  48. 根据权利要求37所述的制备方法，其特征在于：所述方法制备得到的组合物中木糖醇的浓度为1％-5％(m/v)。
  49. 根据权利要求37所述的制备方法，其特征在于：所述方法步骤(1)中防腐剂选自苯酚、邻甲酚、间甲酚、对甲酚、对羟基苯甲酸甲酯、对羟基苯甲酸丙酯、2－苯氧基乙醇、对羟基苯甲酸丁酯、2－苯基乙醇、苯甲醇、氯丁醇、氯甲酚、对羟基苯甲酸乙酯或其混合物，最后得到的组合物浓度为0.1mg/ml到10mg/ml。
  50. 根据权利要求49所述的制备方法，其特征在于：根据所述方法制备得到的组合物中防腐剂的浓度为1mg/ml到8mg/ml。
  51. 根据权利要求50所述的制备方法，其特征在于：根据所述方法制备得到的组合物中防腐剂的浓度为2mg/ml到6mg/ml。
  52. 根据权利要求49-51任一项的制备方法，其特征在于：所述的防腐剂为苯酚或间甲酚。
  53. 根据权利要求52所述的制备方法，其特征在于：根据所述方法制备得到的组合物中防腐剂为苯酚，浓度为0.1mg/ml到10mg/ml。
  54. 根据权利要求53所述的制备方法，其特征在于：根据所述方法制备得到的组合物中防腐剂为苯酚，浓度为1mg/ml到8mg/ml。
  55. 根据权利要求54所述的制备方法，其特征在于：根据所述方法制备得到的组合物中防腐剂为苯酚，浓度为2mg/ml到6mg/ml。
  56. 根据权利要求37所述的制备方法，其特征在于：所述方法步骤(3)组合物中稳定剂选自聚山梨酯20、聚山梨酯80、泊洛沙姆124、泊洛沙姆181、泊洛沙姆182、泊洛沙姆188、泊洛沙姆237、泊洛沙姆331、泊洛沙姆338或泊洛沙姆407。
  57. 根据权利要求56所述的制备方法，其特征在于：所述方法步骤(3)中稳定剂选自聚山梨酯20、聚山梨酯80或泊洛沙姆188，浓度为0.004％至0.3％(m/v)。
  58. 根据权利要求57所述的制备方法，其特征在于：所述方法步骤(3)中稳定剂选自聚山梨酯20、聚山梨酯80或泊洛沙姆188，浓度为0.02％(m/v)。
  59. 根据权利要求58所述的制备方法，其特征在于：所述方法步骤(3)中稳定剂为聚山梨酯80和泊洛沙姆188的混合物，含有0.001％至0.5％(m/v)的聚山梨酯80和0.001％至0.5％(m/v)的泊洛沙姆188。
  60. 根据权利要求59所述的制备方法，其特征在于：所述方法步骤(3)中稳定剂为聚山梨酯80和泊洛沙姆188的混合物，含有0.004％至0.3％(m/v)的聚山梨酯80和0.004％至0.3％(m/v)的泊洛沙姆188。
  61. 根据权利要求60所述的制备方法，其特征在于：所述方法步骤(3)中稳定剂为聚山梨酯80和泊洛沙姆188的混合物，含有0.01％(m/v)的聚山梨酯80和0.01％(m/v)的泊洛沙姆188。
  62. 根据权利要求37所述的制备方法，其特征在于：根据所述方法制备得到的组合物中含有1mg/ml-15mg/ml的利拉鲁肽，pH范围为7.5-8.5、浓度为10-30mmol/L的磷酸氢二钠缓冲剂，0.004％至0.3％(m/v)的聚山梨酯80或泊洛沙姆188，1％-5％(m/v)的木糖醇和1mg/ml-8mg/ml的苯酚或间甲酚。
  63. 根据权利要求37所述的制备方法，其特征在于：根据所述方法制备得到的组合物中含有1mg/ml-15mg/ml的利拉鲁肽，pH范围为7.5-8.5、浓度为10-30mmol/L的磷酸氢二钠缓冲剂，0.004％至0.3％(m/v)的聚山梨酯80和0.004％至0.3％(m/v)的泊洛沙姆188，1％-5％(m/v)的木糖醇和1mg/ml-8mg/ml的苯酚或间甲酚。
  64. 根据权利要求37所述的制备方法，其特征在于：根据所述方法制备得到的组合物中含有3mg/ml-10mg/ml的利拉鲁肽，pH范围为8.0-8.5、浓度为10-30mmol/L的磷酸氢二钠缓冲剂，0.02％(m/v)的聚山梨酯80或泊洛沙姆188，1％-5％(m/v)的木糖醇和2mg/ml-6mg/ml的苯酚或间甲酚。
  65. 根据权利要求37所述的制备方法，其特征在于：根据所述方法制备得到的组合物中含有3mg/ml-10mg/ml的利拉鲁肽，pH范围为8.0-8.5、浓度为10-30mmol/L的磷酸氢二钠缓冲剂，0.01％(m/v)的聚山梨酯80 和0.01％的泊洛沙姆188，1％-5％(m/v)的木糖醇和2mg/ml-6mg/ml的苯酚或间甲酚。
  66. 一种药物组合物，含有0.1mg/ml-25mg/ml的GLP-1类似物，pH7.5-9.0的缓冲剂，0.001％至0.5％(m/v)的稳定剂，0.5％-10％(m/v)的木糖醇和0.1mg/ml-10mg/ml的防腐剂；
    其中，所述的GLP-1类似物为GLP-1或其突变体，GLP-1(7-36)-酰胺或其突变体，GLP-1(7-37)或其突变体，以及上述多肽或其突变体的一个或多个氨基酸残基上引入有机取代基例如酯、烷基或亲脂性官能团化学修饰后的衍生物；
    其中，突变体意指母体肽GLP-1，GLP-1(7-36)-酰胺或GLP-1(7-37)的一个或多个氨基酸残基已经被其他氨基酸残基取代和/或其中母体肽的一个或多个氨基酸残基已经缺失和/或其中已经将一个或多个氨基酸残基添加至母体肽。
  67. 根据权利要求66所述的药物组合物，其特征在于：所述的GLP-1衍生物的亲脂性取代基具有4-40个碳原子，更优选8-30个碳原子，更优选8-25个碳原子，更优选12-25个碳原子，最优选14-18个碳原子。
  68. 根据权利要求66所述的药物组合物，其特征在于：所述的GLP-1类似物为Arg34，Lys26(Nε-(γ-Glu(Nα-十六烷酰)))-GLP-1(7-37)。
  69. 根据权利要求66所述的药物组合物，其特征在于：所述的GLP-1类似物选自：Gly8-GLP-1(7-36)-酰胺，Gly8-GLP-1(7-37)，Val8-GLP-1(7-36)-酰胺，Val8-GLP-1(7-37)，Val8Asp22-GLP-1(7-36)-酰胺，Val8Asp22-GLP-1(7-37)，Val8Glu22-GLP-1(7-36)-酰胺，Val8Glu22-GLP-1(7-37)，Val8Lys22-GLP-1(7-36)-酰胺，Val8Lys22-GLP-1(7-37)，Val8Arg22-GLP-1(7-36)-酰胺，Val8Arg22-GLP-1(7-37)，Val8His22-GLP-1(7-36)-酰胺，Val8His22-GLP-1(7-37)；Arg26-GLP-1(7-37)；Arg34-GLP-1(7-37)；Lys36-GLP-1(7-37)；Arg26， 34Lys36-GLP-1(7-37)；Arg26，34-GLP-1(7-37)；Arg26，34Lys40-GLP-1(7-37)； Arg26Lys36-GLP-1(7-37)；Arg34Lys36-GLP-1(7-37)；Val8Arg22-GLP-1(7-37)；Met8Arg22-GLP-1(7-37)；Gly8His22-GLP-1(7-37)；Val8His22-GLP-1(7-37)；Met8His22-GLP-1(7-37)；His37-GLP-1(7-37)；Gly8-GLP-1(7-37)；Val8-GLP-1(7-37)；Met8-GLP-1(7-37)；Gly8Asp22-GLP-1(7-37)；Val8Asp22-GLP-1(7-37)；Met8Asp22-GLP-1(7-37)；Gly8Glu22-GLP-1(7-37)；Val8Glu22-GLP-1(7-37)；Met8Glu22-GLP-1(7-37)；Gly8Lys22-GLP-1(7-37)；Val8Lys22-GLP-1(7-37)；Met8Lys22-GLP-1(7-37)；Gly8Arg22-GLP-1(7-37)；Val8Lys22His37-GLP-1(7-37)；Gly8Glu22His37-GLP-1(7-37)；Val8Glu22His37-GLP-1(7-37)；Met8Glu22His37-GLP-1(7-37)；Gly8Lys22His37-GLP-1(7-37)；Met8Lys22His37-GLP-1(7-37)；Gly8Arg22His37-GLP-1(7-37)；Val8Arg22His37-GLP-1(7-37)；Met8Arg22His37-GLP-1(7-37)；Gly8His22His37-GLP-1(7-37)；Val8His22His37-GLP-1(7-37)；Met8His33His37-GLP-1(7-37)；Gly8His37-GLP-1(7-37)；Val8His37-GLP-1(7-37)；Met8His37-GLP-1(7-37)；Gly8Asp22His37-GLP-1(7-37)；Val8Asp22His37-GLP-1(7-37)；Met8Asp22His37-GLP-1(7-37)；Arg26-GLP-1(7-36)-酰胺；Arg34-GLP-1(7-36)-酰胺；Lys36-GLP-1(7-36)-酰胺；Arg26， 34Lys36-GLP-1(7-36)-酰胺；Ar26，34-GLP-1(7-36)-酰胺；Arg26， 34Lys40-GLP-1(7-36)-酰胺；Arg26Lys36-GLP-1(7-36)-酰胺；Arg34Lys36-GLP-1(7-36)-酰胺；Gly8-GLP-1(7-36)-酰胺；Val8-GLP-1(7-36)-酰胺；Met8-GLP-1(7-36)-酰胺；Gly8Asp22-GLP-1(7-36)-酰胺；Gly8Glu22His37-GLP-1(7-36)-酰胺；Val8Asp22-GLP-1(7-36)-酰胺；Met8Asp22-GLP-1(7-36)-酰胺；Gly8Glu22-GLP-1(7-36)-酰胺；Val8Glu22-GLP-1(7-36)-酰胺；Met8Glu22-GLP-1(7-36)-酰胺；Gly8Lys22-GLP-1(7-36)-酰胺；Val8Lys22-GLP-1(7-36)-酰胺；Met8Lys22-GLP-1(7-36)-酰胺； Gly8His22His37-GLP-1(7-36)-酰胺；Gly8Arg22-GLP-1(7-36)-酰胺；Val8Arg22-GLP-1(7-36)-酰胺；Met8Arg22-GLP-1(7-36)-酰胺；Gly8His22-GLP-1(7-36)-酰胺；Val8His22-GLP-1(7-36)-酰胺；Met8His22-GLP-1(7-36)-酰胺；His37-GLP-1(7-36)-酰胺；Val8Arg22His37-GLP-1(7-36)-酰胺；Met8Arg22His37-GLP-1(7-36)-酰胺；Gly8His37-GLP-1(7-36)-酰胺；Val8His37-GLP-1(7-36)-酰胺；Met8His37-GLP-1(7-36)-酰胺；Gly8Asp22His37-GLP-1(7-36)-酰胺；Val8Asp22His37-GLP-1(7-36)-酰胺；Met8Asp22His37-GLP-1(7-36)-酰胺；Val8Glu22His37-GLP-1(7-36)-酰胺；Met8Glu22His37-GLP-1(7-36)-酰胺；Gly8Lys22His37-GLP-1(7-36)-酰胺；Val8Lys22His37-GLP-1(7-36)-酰胺；Met8Lys22His37-GLP-1(7-36)-酰胺；Gly8Arg22His37-GLP-1(7-36)-酰胺；Val8His22His37-GLP-1(7-36)-酰胺；Met8His22His37-GLP-1(7-36)-酰胺；Val8Trp19Glu22-GLP-1(7-37)，Val8Glu22Val25-GLP-1(7-37)，Val8Tyr16Glu22-GLP-1(7-37)，Val8Trp16Glu22-GLP-1(7-37)，Val8Leu16Glu22-GLP-1(7-37)，Val8Tyr18Glu22-GLP-1(7-37)，Val8Glu22His37GLP-1(7-37)，Val8Glu22Ile33-GLP-1(7-37)，Val8Trp16Glu22Val25Ile33-GLP-1(7-37)，Val8Trp16Glu22Ile33-GLP-1(7-37)，Val8Glu22Val25Ile33-GLP-1(7-37)，Val8Trp16Glu22Val25-GLP-1(7-37)，及其类似物和这些中任一种的衍生物。
  70. 根据权利要求66所述的药物组合物，其特征在于：所述的药物组合物中，GLP-1类似物的浓度为约1mg/ml-15mg/ml，更优选为3mg/ml-10mg/ml，最优选为6mg/ml。
  71. 根据权利要求66所述的药物组合物，其特征在于：所述的药物组合物中，缓冲剂为能够维持制剂在水溶液状态下pH值为7.5-9.0的任何一种缓冲液，可任选自磷酸缓冲液、磷酸氢二钠-柠檬酸缓冲液、TRIS、甘氨酰基-甘氨酸、N-二(羟乙基)甘氨酸、磷酸二氢钠缓冲液、磷酸氢二钠缓冲液、醋酸钠缓冲液、碳酸钠缓冲液、磷酸钠缓冲液、赖氨酸 缓冲液、精氨酸缓冲液或其混合物。
  72. 根据权利要求71所述的药物组合物，其特征在于：缓冲剂的pH值范围优选为7.5-8.5，更优选为8.0-8.5。
  73. 根据权利要求71所述的药物组合物，其特征在于：缓冲剂的浓度为5-100mmol/L，优选10-30mmol/L。
  74. 根据权利要求73所述的药物组合物，其特征在于：缓冲剂为磷酸氢二钠缓冲液,浓度为5-100mmol/L，pH范围在7.5-8.5之间。
  75. 根据权利要求66所述的药物组合物，其特征在于：所述的药物组合物中，木糖醇的浓度为1％-5％(m/v)。
  76. 根据权利要求66所述的药物组合物，其特征在于：所述的药用组合物中，稳定剂选自氨基酸及衍生物：甘氨酸、丙氨酸、丝氨酸、天冬氨酸、谷氨酸、苏氨酸、色氨酸、赖氨酸、羟赖氨酸、组氨酸、精氨酸、胱氨酸、半胱氨酸、甲硫氨酸、苯丙氨酸、亮氨酸、异亮氨酸等以及它们的衍生物；非离子表面活性剂：脱水山梨醇脂肪酸酯，甘油脂肪酸酯(如脱水山梨糖醇辛酸单酯，脱水山梨糖醇月桂酸单酯和脱水山梨糖醇棕榈酸单酯)，聚甘油脂肪酸酯(例如甘油辛酸单酯，甘油肉豆蔻酸单脂霜和甘油硬脂肪酸单酯)，聚氧乙烯脱水山梨醇脂肪酸酯，聚氧乙烯山梨醇脂肪酸酯，聚氧乙烯甘油脂肪酸酯，聚氧乙烯乙二醇脂肪酸酯，聚氧乙烯烷基醚，聚氧乙烯聚氧丙烯烷基醚，聚氧乙烯苯醚，聚氧乙基化硬蓖麻油，聚氧乙基化蜂蜡衍生物，聚氧乙烯化羊毛脂衍生物或者聚氧乙烯脂肪酸酰胺，阳离子表面活性剂是烷基硫酸盐(如有—个C10—C18烷基的烷基硫酸盐)；聚乙二醇，聚乙烯醇，羟丙基－β－环糊精，羧甲基纤维素，聚乙烯吡咯烷酮,聚山梨酯20或聚山梨酯80，泊洛沙姆系列(泊洛沙姆124、泊洛沙姆181、泊洛沙姆182、泊洛沙姆188、泊洛沙姆237、泊洛沙姆331、泊洛沙姆338或泊洛沙姆407)。
  77. 根据权利要求76所述的药物组合物，其特征在于：所述的稳定剂优选聚山梨酯20、聚山梨酯80、泊洛沙姆124、泊洛沙姆181、泊洛沙姆 182、泊洛沙姆188、泊洛沙姆237、泊洛沙姆331、泊洛沙姆338、泊洛沙姆407或其混合物。
  78. 根据权利要求77所述的药物组合物，其特征在于：所述的稳定剂选自聚山梨酯20、聚山梨酯80、泊洛沙姆188或其混合物。
  79. 根据权利要求66所述的药物组合物，其特征在于：所述的药物组合中，防腐剂选自苯酚、邻甲酚、间甲酚、对甲酚、对羟基苯甲酸甲酯、对羟基苯甲酸丙酯、2－苯氧基乙醇、对羟基苯甲酸丁酯、2－苯基乙醇、苯甲醇、氯丁醇、氯甲酚、对羟基苯甲酸乙酯或其混合物。
  80. 根据权利要求79所述的药物组合物，其特征在于：防腐剂的浓度约为1mg/ml到8mg/ml。
  81. 根据权利要求80所述的药物组合物，其特征在于：防腐剂的浓度约为2mg/ml到6mg/ml。
  82. 一种制备权利要求66所述的GLP-1类似物药物组合物的方法，所述方法包括如下步骤：
    (1)将防腐剂、木糖醇和缓冲剂溶解于注射用水中制得溶液；
    (2)将GLP-1类似物原料溶解于上述溶液中，调节至所需的pH范围；
    (3)在上述溶液中加入稳定剂，得到含有0.1mg/ml-25mg/ml的GLP-1类似物、pH7.5-9.0的缓冲剂、0.001％至0.5％(m/v)的稳定剂、0.5％-10％(m/v)的木糖醇和0.1mg/ml-10mg/ml防腐剂的药物组合物。
PCT/CN2016/081843 2015-05-13 2016-05-12 一种包含glp-1类似物的药物制剂及其制备方法 WO2016180353A1 (zh)

Priority Applications (3)

Application Number Priority Date Filing Date Title
EP16792203.8A EP3295952B1 (en) 2015-05-13 2016-05-12 Pharmaceutical formulation comprising glp-1 analogue and preparation method thereof
US15/571,278 US20190060410A1 (en) 2015-05-13 2016-05-12 Pharmaceutical Formulation Comprising GLP-1 Analogue and Preparation Method Thereof
CN201680007374.3A CN107249620B (zh) 2015-05-13 2016-05-12 一种包含glp-1类似物的药物制剂及其制备方法

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201510243040.3 2015-05-13
CN201510243040 2015-05-13

Publications (1)

Publication Number Publication Date
WO2016180353A1 true WO2016180353A1 (zh) 2016-11-17

Family

ID=57247785

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2016/081843 WO2016180353A1 (zh) 2015-05-13 2016-05-12 一种包含glp-1类似物的药物制剂及其制备方法

Country Status (4)

Country Link
US (1) US20190060410A1 (zh)
EP (1) EP3295952B1 (zh)
CN (1) CN107249620B (zh)
WO (1) WO2016180353A1 (zh)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018096460A1 (en) * 2016-11-22 2018-05-31 Biocon Research Limited Pharmaceutical compositions of glp-1 analogues
WO2018210919A1 (en) * 2017-05-17 2018-11-22 Novo Nordisk A/S Glp-1 compositions and uses thereof
US11590206B2 (en) * 2017-12-21 2023-02-28 Sanofi Liquid pharmaceutical composition

Families Citing this family (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2019506440A (ja) * 2016-03-01 2019-03-07 深▲せん▼翰宇薬業股▲ふん▼有限公司Ｈｙｂｉｏ Ｐｈａｒｍａｃｅｕｔｉｃａｌ Ｃｏ．， Ｌｔｄ． 医薬組成物及びその製造方法
WO2019038412A1 (en) 2017-08-24 2019-02-28 Novo Nordisk A/S GLP-1 COMPOSITIONS AND USES THEREOF
JP2020097531A (ja) * 2018-12-17 2020-06-25 協同乳業株式会社 Ｇｌｐ−１分泌促進剤
GB201917723D0 (en) * 2019-12-04 2020-01-15 Nv Rose Llc Stable liquid formulations of glucagon-like peptide 1 or analogues thereof
IL294520A (en) 2020-02-18 2022-09-01 Novo Nordisk As Pharmaceutical formulations
EP4142695A1 (en) * 2020-04-29 2023-03-08 Novo Nordisk A/S Solid compositions comprising a glp-1 agonist and histidine
WO2022157747A2 (en) * 2021-01-25 2022-07-28 Mylan Ireland Limited Pharmaceutical peptide compositions and methods of preparation thereof
WO2022178737A1 (zh) * 2021-02-25 2022-09-01 杭州九源基因工程有限公司 一种稳定利拉鲁肽药物制剂的处理方法
WO2024017139A1 (zh) * 2022-07-20 2024-01-25 成都海博为药业有限公司 一种含有glp-1受体激动剂类似物的药物组合物

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1195992A (zh) * 1995-08-22 1998-10-14 柏内布拉期加公司 增强肠胃外营养用的方法和组合物
CN1882356A (zh) * 2003-11-20 2006-12-20 诺沃挪第克公司 对于生产和用于注射装置中是最佳的含有丙二醇的肽制剂
US20090286716A1 (en) * 2003-11-13 2009-11-19 Novo Nordisk A/S Soluble Pharmaceutical Compositions for Parenteral Administration Comprising a GLP-1 Peptide and an Insulin Peptide of Short Time Action for Treatment of Diabetes and Bulimia
CN102085355A (zh) * 2011-01-27 2011-06-08 蚌埠丰原涂山制药有限公司 一种利拉鲁肽长效微球注射剂及其制备方法
CN103405753A (zh) * 2013-08-13 2013-11-27 上海华谊生物技术有限公司 稳定的促胰岛素分泌肽水针药物组合物
WO2014030051A1 (en) * 2012-08-23 2014-02-27 Aurobindo Pharma Limited Stable pharmaceutical compositions comprising saxagliptin
CN103893744A (zh) * 2012-12-24 2014-07-02 杭州九源基因工程有限公司 一种治疗糖尿病的药物制剂及其制备方法
CN104548096A (zh) * 2013-10-09 2015-04-29 深圳翰宇药业股份有限公司 一种含有glp-1类似物和dpp-4抑制剂的药物组合物及其制备方法

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1412384B1 (en) * 2001-06-28 2007-12-26 Novo Nordisk A/S Stable formulation of modified glp-1
CN1812807A (zh) * 2003-06-03 2006-08-02 诺沃挪第克公司 稳定化的药物肽组合物
US20080125361A1 (en) * 2004-11-12 2008-05-29 Novo Nordisk A/S Stable Formulations Of Peptides

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1195992A (zh) * 1995-08-22 1998-10-14 柏内布拉期加公司 增强肠胃外营养用的方法和组合物
US20090286716A1 (en) * 2003-11-13 2009-11-19 Novo Nordisk A/S Soluble Pharmaceutical Compositions for Parenteral Administration Comprising a GLP-1 Peptide and an Insulin Peptide of Short Time Action for Treatment of Diabetes and Bulimia
CN1882356A (zh) * 2003-11-20 2006-12-20 诺沃挪第克公司 对于生产和用于注射装置中是最佳的含有丙二醇的肽制剂
CN102085355A (zh) * 2011-01-27 2011-06-08 蚌埠丰原涂山制药有限公司 一种利拉鲁肽长效微球注射剂及其制备方法
WO2014030051A1 (en) * 2012-08-23 2014-02-27 Aurobindo Pharma Limited Stable pharmaceutical compositions comprising saxagliptin
CN103893744A (zh) * 2012-12-24 2014-07-02 杭州九源基因工程有限公司 一种治疗糖尿病的药物制剂及其制备方法
CN103405753A (zh) * 2013-08-13 2013-11-27 上海华谊生物技术有限公司 稳定的促胰岛素分泌肽水针药物组合物
CN104548096A (zh) * 2013-10-09 2015-04-29 深圳翰宇药业股份有限公司 一种含有glp-1类似物和dpp-4抑制剂的药物组合物及其制备方法

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of EP3295952A4 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018096460A1 (en) * 2016-11-22 2018-05-31 Biocon Research Limited Pharmaceutical compositions of glp-1 analogues
WO2018210919A1 (en) * 2017-05-17 2018-11-22 Novo Nordisk A/S Glp-1 compositions and uses thereof
US11590206B2 (en) * 2017-12-21 2023-02-28 Sanofi Liquid pharmaceutical composition

Also Published As

Publication number Publication date
CN107249620A (zh) 2017-10-13
CN107249620B (zh) 2018-06-26
EP3295952A4 (en) 2018-12-12
EP3295952A1 (en) 2018-03-21
EP3295952B1 (en) 2021-02-17
US20190060410A1 (en) 2019-02-28

Similar Documents

Publication Publication Date Title
EP3295952B1 (en) Pharmaceutical formulation comprising glp-1 analogue and preparation method thereof
US20220016217A1 (en) Combination of an insulin and a glp-1-agonist
EP1478233B1 (en) Method for administering glp-1 molecules
US7179788B2 (en) Biphasic mixtures of GLP-1 and insulin
AU765584B2 (en) Protein formulations
US7238663B2 (en) Pre-mixes of GLP-1 and basal insulin
RU2440097C2 (ru) Способ лечения диабета ii типа и ожирения, осмотическое устройство для доставки и способ его изготовления
WO2022111370A1 (zh) 一类glp-1/胰高血糖素受体双重激动剂及其应用
EP1412384B1 (en) Stable formulation of modified glp-1
JP5675799B2 (ja) 遅効性インスリン製剤
EP2219665B1 (en) Parathyroid hormone formulations and uses thereof
CA2487585A1 (en) Formulations for amylin agonist peptides
TW201100093A (en) Preparation comprising insulin, nicotinamide and an amino acid
JP2013510821A (ja) Ｇｌｐ−１アゴニスト及びメチオニンを含む薬学的組成物
TW201105346A (en) Heat-stable and vibration-stable insulin preparations
CN103893744B (zh) 一种治疗糖尿病的药物制剂及其制备方法
EP3257524B1 (en) Stabilized solution preparation of pharmaceutical glp-1r antibody fusion protein
CN114053217B (zh) 一种Exendin-4-Fc融合蛋白注射制剂及其制备方法
JP2009149684A (ja) アミリン作動薬ペプチド用製剤
AU2002313662A1 (en) Pre-mixes of GLP-1 and basal insulin

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: 16792203

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE