CN103861089B - The derivative of GLP-1 analog or its officinal salt injection and its preparation method and application - Google Patents

Abstract

The present invention relates to the derivatives of GLP-1 analog or its officinal salt injection and its preparation method and application.Specifically, the invention discloses the derivative of GLP-1 analog or the injection of its officinal salt and its preparation method and application shown in a kind of formula (I), contain the derivative of GLP-1 analog shown in 10~50mg formula (I) or its considerable amount of officinal salt, 5~15mg hydroxypropyl-β-cyclodextrin, 30~60mg NaCl and water for injection in every milliliter of injection.The derivative of GLP-1 analog shown in formula (I) provided by the present invention or its officinal salt injection are with good stability, and prescription is simple, are conducive to industrialized production.His-Aib-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu- Glu-Glu-Glu-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-Leu-Val-Arg- Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-Lys(I).

Description

The derivative of GLP-1 analog or its officinal salt injection and preparation method thereof and Purposes

Technical field

The present invention relates to the derivative of GLP-1 analog shown in a kind of formula (I) or the injection of its officinal salt and its systems Preparation Method and purposes.

Technical background

Diabetes are a kind of global prevalence diseases, are grape absolutely or caused by relative deficiency due to internal insulin Syndrome (present Research Guangdong Pharmaceutical University of Chen Ruijie Remedies for diabetes of sugar, protein, disorders of lipid metabolism Report, 2001,7 (2): 131-133), I type and type II diabetes (Type2diabetes can be divided into according to pathogenesis Mellitus, T2DM, similarly hereinafter).In all diabetics made a definite diagnosis, patient 90-95% suffers from T2DM, and patient is often Along with fat, physical activity deficiency (physical inactivity), older, family's diabetic history, glucose metabolism Damage and have family's diabetic history etc..T2DM is also a kind of progressive disease (progressive disease).2000, There are about 1.71 hundred million people to suffer from diabetes in World Health Organization's statistical data estimation whole world；2005, U.S.'s disease control and prevention There are about 0.208 hundred million Americans to suffer from diabetes for center (Centers for Disease compares and Prevention) estimation, about Account for the 7% of U.S. population；It is counted according to International Diabetes Federation within 2006, global diabetes number of patients, which is about 2.46 hundred million, (to be accounted for about complete The 5.9% of ball total population) and 46% patient age between 40-59 years old.Studies have shown that normal person and T2DM patient are to grape Sugar reaction has very important difference.Normal person belongs to early stage insulin response (early to the reaction of blood glucose rise after the meal Insulin response).

T2DM feature is that insulin inhibits and pancreas Instreptozotocin Induced damages, and leads to insulin deficit and hyperglycemia (Ferrannini E.Insulin resistance versus insulin deficiency in non-insulin- dependent diabetes mellitus:problems and prospects.Endocr Rev.1998,19(4):477– 490).Patient T2DM generally will appear after meal and hyperglycemia (fasting blood-glucose > 125mg/dL), and hyperglycemia is mainly As pancreas beta cell cannot secrete enough insulin compensate the insulin in perienchyma inhibit caused by (Weyer C.,Bogardus C.,Mott DM.,et al.The natural history of insulin secretory dysfunction and insulin resistance in the pathogenesis of type2diabetes Mellitus.J.Clin.Invest.1999,104 (6): 787-794).

The Major Risk Factors of T2DM are obesities, it has very big harm to human health.Patient suffers from angiocarpy The danger of disease and unusual death can become larger, at the same T2DM often with some other high risk diseases, such as hypertension, blood lipid Obstacle and obesity etc. exist simultaneously；60% patient T2DM includes retinopathy along with microvascular complication person (retinopathy) and neuropathy (neuropathy) and cardiovascular pathologies (cardiovascular related with T2DM Morbidities) such as coronary heart disease, myocardial infarction and shock.In the U.S., cardiovascular disease (cardiovascular disease, CVD) is to lead to morbidity and dead principal element, and type-2 diabetes mellitus is big blood vessel (macrovascular) complication such as atherosclerosis (atherosclerosis), myocardial infarction (myocardial Infarction), the Major Risk Factors (major risk factor) of the generation of shock and peripheral vascular disease.Suffer from diabetes The adult heart disease and shock of occurring cause dead probability to be 2-4 times of non-diabetes patient, in addition, close to 65% diabetes People dies of heart disease and shock.

Other than the injury to patient's physiology and body, T2DM can also cause very big financial burden to society, according to system Meter, is used to treat about 22,900,000,000 dollars of expense of diabetic complication every year in the U.S., annual for T2DM and its complication For total cost close to 57,100,000,000 dollars, out-of-pocket cost total value is more than 8,000,000,000 dollars.

T2DM therapeutic agent is always focus concerned by people, from the sulphonyl class of early stage, biguanides oral hypoglycemic drug to Recent insulin sensitizer and alpha-glucosidase inhibitor, from animal insulin to actrapid monotard and the exploitation of various novel forms, From the drug therapy mechanism of simple increase insulin to the new way for acting on generation insulin earlier.Weight gain is Common adverse reaction after this kind of oral or injection antidiabetic drug medication, this point may be decreased compliance, and can increase hair The risk of raw cardiovascular disease.Therefore develop highly-safe, patient's biddability is good, the low novel T2DM therapeutic agent of adverse reaction at Fall over each other the hot spot developed for numerous research institutions and pharmacy corporation.

Before more than 100 years, Moore just proposes that duodenum can be secreted " the chemical excitant " of stimulating pancreas secretion, and Attempt to inject enteron aisle extract to treat diabetes.The humoral factor for being subsequently found intestinal secretion can enhance the endocrine function of pancreas Can, no matter the insulin secretion that vein or oral glucose are caused, the stimulation of about 50% peptides caused by the enteron aisle makees With Zunz and Labarre proposes the concept of " duodenin (incretin) " thus.So far separated 2 kinds of intestines out are hypoglycemic Element, i.e. glucose dependent insulin release peptide (glucose-dependent insulinotropic polypeptide, ) and glucagon-like-peptide-1 (glucagon-like peptide-1, GLP-1) GIP.GIP and GLP-1 is inhaled in nutrition Time receiving is secreted by specific enteric nervous secretory cell, and wherein GIP is by duodenum and neighbouring jejunum K cell (proximal Jejunal K cells) it secretes, GLP-1 is then synthesized in L cell and is primarily present in distal small bowel and colon (Drucker DJ.Enhancing incretin action for the treatment of type2diabetes.Diabetes Care.2003,26 (10): 2929-2940).

GLP-1 exists in blood plasma with two kinds of biologically active forms of GLP-1 (7-37) and GLP-1 (7-36) amide, this two A polypeptide only has an amino acid of differences, and they biological effect and Half-life in vivo be identical (Drucker DJ.Enhancing incretin action for the treatment of type2diabetes.Diabetes Care.2003,26 (10): 2929-2940).

Usually said GLP-1 is the general designation to GLP-1 (7-37) and GLP-1 (7-36) amide.GIP and GLP-1 are in stomach Enteron aisle can be degraded by intracorporal dipeptidyl peptidase-IV (dipeptidyl peptidase-IV, DPP-IV) quickly when releasing For inactive form, thus make the half-life period of GIP and GLP-1 in vivo it is very low (GIP Half-life in vivo about 5-7 minutes, GLP-1 Half-life in vivo about 2 minutes) (Drucker DJ.Enhancing incretin action for the Treatment of type2diabetes.Diabetes Care.2003,26 (10): 2929-2940).Studies have shown that mostly Number degradation processes occur to occur when GIP and GLP-1 enters blood vessel containing DPP-IV, the GLP-1 not being degraded on a small quantity and GIP can enter pancreas and with its binding site combine and stimulate beta cell release insulin.With sulfonylureas (sulfonylureas) Insulin mechanism difference is discharged by directly facilitating function beta cell, it is all dependence on the glucose that incretin effect is most of Type.In addition, some animals and human vitronectin are experiments have shown that also there are GLP-1 A cells to inhibit and reduce glucagon excessively to divide The effects of secreting (glucagon hypersecretion).

Although T2DM patient's body blood plasma GIP level is normal but the effect of its incretin is remarkably decreased or loses, and GLP-1 is then to reduce, therefore study the drug based on GLP-1 and be more conducive to treatment T2DM in T2DM patient's body level.Although GLP-1 (7-37) and GLP-1 (7-36) amide level can improve in a few minutes after meal, and GLP-1 (7-36) amide content is more A bit, thus endocrine and nerve signal transmitting double action may the food being digested from alimentary canal lower end enter small intestine and The secretion of GLP-1 has been substantially increased before colon.GLP-1 level is very low (about 5-10pmol/ in blood plasma under fasting state L), and its level increases sharply and (reaches 15-50pmol/L) after feed.In the case where DPP-IV and kidney remove double action, in vivo The GLP-1 level of circulation reduces rapidly, and other enzymes such as human body neutral endopeptidase 24.11(human neutral ) etc. endopeptidase2411 also whether the research work for having lost conclusive effect active to GLP-1 is carrying out In.Since GLP-1 is the good substrate that alanine is DPP-IV in 2 amino acids, it is easier to be degraded to inactive peptide fragment. In fact, the main reason for DPP-IV is only incretin loss of activity in vivo, experiment shows what DPP-IV gene was silenced GLP-1 level significantly improves in Mice Body, increases the secretion of insulin.Exactly under DPP-IV effect, complete in vivo and tool The GLP-1 for having bioactivity is only 10-20%(Deacon CF, the Nauck MA, Toft- of blood plasma GLP-1 total content Nielsen M,et al.Both subcutaneously and intravenously administered glucagon- like peptide1are rapidly degraded from the NH2-terminus in type2-diabetic Patients and in healthy subjects.Diabetes.1995,44 (9): 1126-1131).

GLP-1 and GIP passes through the G- G-protein linked receptor (G-protein-coupled entirely different with structure Receptors, GPCRs) in conjunction with and play each self-applying.GIP receptor is expressed by pancreaticβ-cell, and small part is in rouge Fat tissue and central nervous system expression.In contrast, GLP-1 receptor is then mainly in pancreatic islet alpha-and beta cell and periphery group It knits including being expressed in maincenter and peripheral nervous system, brain, kidney, lung and gastrointestinal tract etc..Two incretin are in beta cell Activation will lead to increasing sharply for cAMP and intracellular calcium levels, so as to cause its with glucose-dependent manner to exocytosis, hold Continuous incretin receptor signal transmitting will lead to genetic transcription, the biosynthesis for increasing insulin with protein kinase A correlation And stimulate proliferation (the Gallwitz B.Glucagon-like peptide-1-based therapies for of beta cell The treatment of type2diabetes mellitus.Treat Endocrinol.2005,4 (6): 361-370). The activation of GLP-1 and GIP receptor can also inhibit beta cell apoptosis in rodent and Human islet while increase its survival Rate (Li Y, Hansotia T, Yusta B, et al.Glucagon-like peptide-1receptor signaling Modulates beta cell apoptosis.J Biol Chem.2003,278 (1): 471-478).With GLP-1 by body surface Up to it is consistent be GLP-1 can also be secreted with glucagon suppression, gastric emptying and food intake, while passing through neuromechanism (neural mechanism) enhances the degradation to glucose.It should be noted that react identical with other insulin secretions, GLP-1 is glucagon dependence to the effect of glucose production, and causes counter regulation to discharge since hypoglycemia acts on (counter-regulatory release of glucagon) though glucagon act in GLP-1 pharmacological concentrations Under be still fully retained.

Endogenous GLP-1 and GIP is short of money in receptor for the important physiological role played in glucose homeostasis It is had made intensive studies in anti-agent or knock out mice.The antagonism of acute GLP-1 or GIP reduces rodent body The secretion of interior insulin and increase Plasma glucose level.Equally, GIP or GLP-1 receptor deactivation mutant mice equally has The secretion of deficiency glucose stimulated insulin and damaging glucose tolerance.GLP-1 also has fasting blood-glucose regulatory function, because It will lead to the raising of rodent fasting glucose level for the acute antagonism or genetic disruption of GLP-1 effect；Meanwhile GLP- 1 is the basis that glucose controls in human body, to antagonism Exendin's (9-39) research shows that GLP-1 effect can make after being destroyed At defective glucose stimulated insulin secretion (defective glucose-stimulated insulin secretion), Glucose clearance is reduced, Plasma Glucagon Level is increased and accelerates gastric emptying.In addition, the physiological action of GLP-1 (Deacon CF.Therapeutic strategies based on glucagon-like Peptide1.Diabetes.2004,53 (9): 2181-2189) further include: (1) help tissue blood glucose absorb, mediating glucose according to Rely type insulin secretion；(2) inhibit postprandial glucagon secretion, reduce glycogen release；(3) gastric emptying is adjusted, prevents food from existing The excessive circulation of glucose when intestinal absorption；(4) food intake (such as appetite) is inhibited.Furthermore animal experiment also shows the one of GLP-1 A physiological role is pancreas beta cell quantity in liptinite.

Because GLP-1 and GIP is many-sided with good action in control blood glucose etc., especially it does not generate hypoglycemia and prolongs The characteristics of slow gastric emptying control weight, causes the interest of numerous scientists.People begin trying research based on GLP-1 and GIP medicine Object is for treating T2DM.It is well known that patient T2DM lacks or loses incretin effect, one reason for this is that T2DM The incretin effect of GIP weakens significantly in patient body；Meanwhile T2DM patient's body GLP-1 level is very low, and because diet pierces Swash the GLP-1 level generated and substantially reduces (Toft-Nielsen MB, Damholt MB, Madsbad S, et al.Determinants of the impaired secretion of glucagon-like peptide-1in Type2diabetic patients.J Clin Endocrinol Metab.2001,86 (8): 3717-3723).Because of T2DM Patient's body GLP-1 acts on having obtained part reservation, it is intended to enhance the drug research of T2DM patient's body incretin effect One of direction is exactly GLP-1 synergist.

GLP-1 analog can stimulate internal pancreas islet as endogenous GLP or GIP in a manner of dependence on the glucose type The secretion of element, while the internal release of glucagon suppression.Furthermore GLP-1 analog has effect to following symptom: (1) low Blood glucose.Different from other rush secretion drug, GLP-1 analog is due to promoting internal insulin point with a kind of glucose-dependent manner It secretes, therefore its hypoglycemic effect has self limiting, and serious hypoglycemia will not be generally caused in large dosage.In spite of document report Accuse GLP-1 and blood glucose can be reduced to normal level hereinafter, but the effect it is of short duration, and be considered as GLP-1 pancreotropic hormone point Secrete the natural result of effect.Since the inactivation of insulin needs certain time, reduced when because of blood sugar concentration, the thorn of GLP-1 When swashing declines without neo-insulin secretion, original insulin is still in action.In short, GLP-1 can make blood Sugar is temporarily reduced under normal level, but not causes serious and lasting hypoglycemia.(2) to being satiated with food and the effect of weight.It removes It directly reduces except blood glucose, GLP-1 can also reduce the intake of food, this is all obtained on rodent and the person Verifying.In this way blood glucose level can be indirectly controlled by losing weight.GLP-1 inhibits gastrin and feed stimulation there are also potential Gastric acid secretion effect, these effects show that GLP-1 may also have the function of prevention of digestive tract ulcers.The effect of GLP-1 Mechanism makes it that can not only become ideal diabetes B, can also become the therapeutic agent of obese diabetic patient.GLP- 1 can be enhanced the satietion of patient, reduces food intake and keep weight or weight-reducing；(3) beta cell health is maintained.Some researchs Prompt GLP-1 can prevent by the conversion of impaired glucose tolerance to diabetes, and there are also some reported literatures GLP-1 class compounds pair The growth of experimental animal beta Cell of islet and proliferation have direct effect, and have experiment discovery GLP-1 that can promote pancreatic stem Differentiation of the cell to functional beta cells.These results imply that GLP-1 has the function of protecting pancreas islet and delays diabetes development, The form and function of beta cell can be kept, while reducing its apoptosis；(4) to the effect of postprandial hyperglycemia.This phenomenon represents A kind of new direction of T2DM treatment.Since some oral drugs and exogenous insulin cannot inhibit or reduce the high blood of T2DM patient's pancreas Sugared element crosses hypersecretion, and GLP-1 analog can may be discharged by direct glucagon suppression or because promoting insulin point Secrete and the paracrine inhibiting effect that generates and glucagon hypersecretion is had an impact.Can effectively it be subtracted by the two mechanism Few postprandial hyperglycemia phenomenon；Meanwhile keeping Instreptozotocin Induced that there may also be effect to long-term control postprandial hyperglycemia.

GLP-1 analog is taken by subcutaneous injection simultaneously, and the amount for not needing to calculate carbon water compound is best to estimate Drug dose, do not need yet to blood glucose carry out self monitor, keep the use of this kind of drug more more convenient than insulin.

The treatment for turning out to be diabetes B of natural GLP-1 multiple efficacies brings new hope, and still, human body is natural GLP-1 is very unstable, can be degraded by DPP IV (DPP-IV), half-life period is only 1~2 minute.According to natural GLP-1 Blood glucose is reduced, needs Intravenous Infusion or continuous subcutaneous infusion, Clinical feasibility is poor.Faced with this situation, people are continuous It explores, it would be desirable to find the method for extending GLP-1 action time.Therefore long-acting GLP-1 analog or derivatives thereof is developed, at For the key areas of the world of medicine's concern.

Exenatide is the Exendin-4 of synthesis, is developed cooperatively by Li Lai company and Amylin company, trade nameFDA and EMEA has been approved by its listing, for treating T2DM.It has 50% with mammal GLP-1 in sequence Homology and its (Drucker DJ, Nauck MA.The incretin similar with GLP-1 to the affine site of GLP-1 receptor system:glucagon-like peptide-1receptor agonists and dipeptidyl peptidase- 4inhibitors in type2diabetes.Lancet.2006,368 (9548): 1696-1705), by the distinctive base of lizard Because of coding；Compared with GLP-1, the 2nd alanine of GLP-1 is substituted for glycine by Exenatide, effectively inhibits DPP-IV Enzymatic hydrolysis, partly decline its about 60-90 minutes (Kolterman OG, Kim DD, Shen L, et in vivo al.Pharmacokinetics,pharmacodynamics,and safety of exenatide in patients with Type2diabetes melllitus.Am Health Syst Pharm.2005,62 (2): 173-181), single subcutaneous injection Concentration continues to increase Exenatide in vivo afterwards, maximum concentration can be reached in blood plasma within 2 hours or so, can maintain 4-6 hours (Nielsen LL, Baron AD.Pharmacology of exenatide (synthetic exendin-4) for the Treatment of type2diabetes.Curr Opin Investig Drugs.2003,4 (4): 401-05).It needs to infuse Meaning is that the metabolism of Exenatide does not occur in liver, and mainly through proteasome degradation after glomerular filtration.

Exenatide have special glucose modulatory activities, including dependence on the glucose type enhancing insulin secretion effect, The effects of dependence on the glucose type inhibits incorrect excessively high glucagon secretion effect, slows down gastric emptying and reduce food intake. To the research of internal and external diabetes model, it has also been found that, Exenatide also has storage first stage (first-phase) pancreas islet Element secretion promotes beta-cell proliferation and insulin from the regenerated effect of its precursor.

In order to reach preferable glycemic control, double injection Exenatide is needed daily, this brings very big to patient It is inconvenient.Furthermore also there is Exenatide slight extremely medium gastric disorder causing nausea (about 40% patient has this reaction), diarrhea and vomiting (to be less than 15% patient has both reactions)；About 50% can generate antibody in the patient of Exenatide treatment, although these antibody are not It will affect drug effect or cause other clinical effects.After taking Byetta bleeding or necrotizing pancreatitis occur for discovery 6 again recently Symptom.

CJC-1131 is that a kind of peptase suppressive GLP-1 of ConjuChem Biotechnologies Inc exploitation is similar 2 in GLP-1 sequence Ala have been substituted for D-Ala by object, to enhance the ability for resisting DPPIV enzymatic hydrolysis, are wrapped in structure Containing one there is reactivity linking agent (reactive linker) to be integrated to blood by covalent (non-reversible) mode in order to it (Kim JG, Baggio LL, Bridon DP, et al.Development and characterization on pure albumen of a glucagon-like peptide-1albumin conjugate:the ability to activate the Glucagon-like peptide1receptor in vivo.Diabetes.2003,52 (3): 751-759), generation GLP-1- seralbumin compound remains the activity of GLP-1, while increasing and digesting stability to DPP-IV, extends body Interior action time, plasma clearance half-life period about 20 days.

One carried out is the study found that CJC-1131- seralbumin compound and use human recombinant pancreas GLP-1 The K that Ki is about 12nM(GLP-1 when the Chinese hamster ovary cell of receptor transfection combines_iFor 5.2nM)；This is complex activating simultaneously The EC50 of cAMP is 11-13nM, EC₅₀It is similar to GLP-1.Existing literature show the binding molecule can reduce blood glucose normally and Hyperglycemia mouse postprandial blood sugar concentration, and experiments have shown that CJC-1131 this active function in GLP-1 it is a certain it is functional by On body, while CJC-1131 also has the effects that slow down gastric emptying and inhibits food intake in mouse.

CJC-1131 has been completed part ii clinical trial phase.In September, 2005, ConjuChem tie existing test Think that CJC-1131 may be not suitable for chronic dose strategy (chronic dosing regimens) after fruit analysis, thus suspends The clinical research of CJC-1131.CJC-1131 clinical test at present does not restart yet.

Albugon(albumin-GLP-1) be by GlaxoSmithKline PLC Human Genome Sciences Inc authorization under A kind of long-acting T2DM therapeutic agent of exploitation, it is the fusion of GLP-1 (with the mutation increased to DDP-IV resistance) and albumin Body.It is 3 days in monkey intracorporal half-life period.Its basic Research idea is after recombinating GLP-1 and seralbumin coupling The compound formed, just significantly increases its Half-life in vivo in this way.Mouse blood sugar is effectively reduced after taking Albugon Horizontal, the effects of increasing insulin secretion, slow down gastric emptying and reducing food intake (Baggio LL, Huang Q, Brown TJ,et al.A Recombinant Human Glucagon-Like Peptide(GLP)-1-Albumin Protein(Albugon)Mimics Peptidergic Activation of GLP-1Receptor-Dependent Pathways Coupled With Satiety,Gastrointestinal Motility,and Glucose Homeostasis.Diabetes.2004,53 (9): 2492-2500).Albugon is carrying out III clinical trial phase at present.

WO9808871 discloses a kind of GLP-1 derivative that fatty acid modifying is carried out on the basis of GLP-1 (7-37), So that the half-life period of GLP-1 in vivo is greatly enhanced.WO9943705, which discloses a kind of N-terminal in GLP-1, to carry out chemistry and repairs The derivative of decorations, but there is document report to carry out modification in the amino acid of N-terminal and will cause the activity of entire GLP-1 derivative significantly It reduces (J.Med.Chem.2000,43,16641669).In addition CN200680006362, CN200680006474, The patents such as WO2007113205, CN200480004658, CN200810152147, W O2006097538 also disclose a series of GLP-1 analog obtained through chemical modification or amino acid substitution or derivatives thereof, wherein most representational is NovoNorm The liraglutide of moral company exploitation has been completed III phase clinic.Liraglutide is a kind of GLP-1 derivative, knot There is the GLP-1 analog of 97% homology containing sequence and source of people GLP-1, the analog and palmitinic acid are covalently attached structure in structure At Liraglutide^[, the palmitinic acid in Liraglutide structure is connected on seralbumin with non covalent forms, this Structure feature determines the slave injection site release that it will be slow, and extends on the basis of not changing its GLP-1 activity and follows in vivo Ring half-life period；The palmitinic acid in structure will form certain space steric hindrance simultaneously, to prevent the degradation of DPP-IV, reduce Kidney scavenging effect.Due to the above characteristic, Liraglutide after through subcutaneous injection people's intracorporal half-life period about It 10-14 hours, can be theoretically administered once with one day, daily dosage is 0.6-1.8mg.

Polypeptide drug is widely used at present in clinical research or therapeutic process.It is stable, high-quality, can produce The therapeutical peptide class pharmaceutical preparation that industry metaplasia produces, for researcher or a great challenge.Polypeptide drug The stability of preparation includes: two broad aspect of chemical stability and physical stability；The factor for influencing its chemical stability is mainly The variation of covalent bond, such as hydrolysis, deamination, oxidation, racemization or crosslinked action；Influence physically stable sexual factor include denaturation, aggregation, Absorption or precipitating etc..

The derivative of GLP-1 analog shown in formula (I) or its officinal salt are a kind of new compounds, are treating non-pancreas islet Curative effect is obvious in terms of the diseases such as plain dependence or insulin-dependent diabetes mellitus or treatment obesity, CN200910165559.9, The patent documents such as WO2011012080A1 have relevant teachings.Currently, it is good to need to develop a kind of stability in the market, preparation process Simply, the derivative of GLP-1 analog shown in formula (I) easy to use or its officinal salt injection.

Summary of the invention

It is an object of the invention to the derivative of GLP-1 analog shown in a kind of formula (I) or its officinal salt injection, In in every milliliter of injection containing the derivative of GLP-1 analog shown in 10~50mg formula (I) or its considerable amount of officinal salt, 5~15mg hydroxypropyl-β-cyclodextrin, 30~60mg NaCl and water for injection.

His-Aib-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Glu-Glu- Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-Leu-Val-Arg-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro- Pro-Pro-Ser-Lys

(I)

In an optimal technical scheme of the invention, the derivative of GLP-1 analog shown in formula (I) or its officinal salt are 20~40mg, more preferable 30mg.

In an optimal technical scheme of the invention, hydroxypropyl-β-cyclodextrin 10mg.

Preferably, the weight ratio of GLP-1 analog and hydroxypropyl-β-cyclodextrin is 2:1 in injection of the present invention.

In an optimal technical scheme of the invention, using sodium hydroxide or salt acid for adjusting pH to 4~6.

In an optimal technical scheme of the invention, sodium hydroxide 5mg.

In an optimal technical scheme of the invention, NaCl is 40~50mg, more preferable 45mg.

The method for preparing the derivative or its officinal salt injection of GLP-1 analog shown in formula (I), including walk as follows It is rapid:

1) derivative of GLP-1 analog shown in formula (I) or its officinal salt are dissolved with injection solvent；

2) sodium chloride and hydroxypropyl-β-cyclodextrin is added, dissolves by heating；

3) sodium hydroxide or hydrochloric acid is added, adjusting pH value is 4~6；

4) water for injection constant volume is used, is filtered, bottling.

In one optimal technical scheme of preparation method of the present invention, solvent for injection is selected from ethyl alcohol, propylene glycol, poly- second two Alcohol, benzyl alcohol and glycerol.

In one optimal technical scheme of preparation method of the present invention, pH value 5.

It is another object of the present invention to provide the derivative of GLP-1 analog shown in a kind of formula (I) or its is pharmaceutically acceptable The injection of salt is in preparation for treating non-insulin-dependent or insulin-dependent diabetes mellitus or treating the drug of obesity In purposes.

The derivative of GLP-1 analog shown in formula (I) provided by the present invention or its officinal salt injection have good Stability, and prescription is simple, is conducive to industrialized production.

Specific embodiment

The present invention will be further illustrated by specific embodiment below, but this does not represent the present invention and is only defined in tool The express ranges of body embodiment.

Embodiment 1

Prescription (based on 1000 bottles):

Preparation process:

By recipe quantity, the derivative of GLP-1 analog shown in formula (I) is added in 20% ethanol solution 1000ml, dissolution And stir evenly, sodium chloride and hydroxypropyl-β-cyclodextrin are added, is dissolved by heating, sodium hydroxide and appropriate hydrochloric acid tune is added PH to 5 is saved, with water for injection constant volume.Solution is sent into desinfection chamber, the filtering with microporous membrane through 0.22um is to clarifying, by every bottle The loading amount of 6ml is filling in the cillin bottle of 25ml, beyond the Great Wall butyl rubber bung.

Embodiment 2

Prescription (based on 1000 bottles):

Preparation process:

By recipe quantity, the derivative of GLP-1 analog shown in formula (I) is added in 20% benzyl alcohol solution 1200ml, it is molten It solves and stirs evenly, add sodium chloride and hydroxypropyl-β-cyclodextrin, dissolve by heating, sodium hydroxide is added and adjusts pH to 5, uses Water for injection constant volume.Solution is sent into desinfection chamber, the filtering with microporous membrane through 0.22um is to clarifying, by the loading amount of every bottle of 6ml It is filling in the cillin bottle of 25ml, butyl rubber bung beyond the Great Wall.

Embodiment 3

Prescription (based on 1000 bottles):

Preparation process:

By recipe quantity, the derivative of GLP-1 analog shown in formula (I) is added in 40% glycerol 800ml, dissolves and stir It mixes uniformly, adds sodium chloride and hydroxypropyl-β-cyclodextrin, dissolve by heating, sodium hydroxide is added and adjusts pH to 5, uses injection Water constant volume.Solution is sent into desinfection chamber, the filtering with microporous membrane through 0.22um to clarify, by the loading amount of every bottle of 6ml it is filling in In the cillin bottle of 25ml, butyl rubber bung beyond the Great Wall.

Embodiment 4

Prescription (based on 1000 bottles):

Preparation process:

By recipe quantity, the derivative of GLP-1 analog shown in formula (I) is added in 30% propylene glycol solution 1500ml, it is molten It solves and stirs evenly, add sodium chloride and hydroxypropyl-β-cyclodextrin, dissolve by heating, sodium hydroxide and appropriate hydrochloric acid is added PH to 5 is adjusted, with water for injection constant volume.Solution is sent into desinfection chamber, the filtering with microporous membrane through 0.22um is to clarifying, by every The loading amount of bottle 6ml is filling in the cillin bottle of 25ml, beyond the Great Wall butyl rubber bung.

Embodiment 5

Prescription (based on 1000 bottles):

Preparation process:

By recipe quantity, the derivative of GLP-1 analog shown in formula (I) is added in 20% polyglycol solution 900ml, It dissolves and stirs evenly, add sodium chloride and hydroxypropyl-β-cyclodextrin, dissolve by heating, sodium hydroxide and appropriate salt is added Acid for adjusting pH is to 5, with water for injection constant volume.Solution is sent into desinfection chamber, the filtering with microporous membrane through 0.22um is pressed to clarifying The loading amount of every bottle of 6ml is filling in the cillin bottle of 25ml, beyond the Great Wall butyl rubber bung.

1 stability experiment of experimental example

Experimental method: it takes the sample of each embodiment of 1ml to be placed in special centrifuge tube respectively, is put into table model high speed centrifuge Centrifuge tube is taken out after centrifugation 15 minutes with the centrifugation of 2000rpm revolving speed.It is by bottom end that sample instillation small beaker is appropriate, it is taken with micro Sample device is drawn 50.0 μ L and is added in 25ml measuring bottle, is diluted to scale with water for injection, mixes.With water level blank in 500nm wavelength Under, it detects it and absorbs angle value (A).50.0 μ L raw samples are taken to be placed in 25ml measuring bottle again, with water for injection constant volume, in Same Wavelength Place's detection its absorption value (A₀).By mathematical formulae K_E=(| A₀-A|/A₀) * 100%, calculate the stability parameter K of the emulsion_E。K_EMore Small emulsion is more stable.Experimental result is as shown in table 1:

1 stability experiment result of table

Sample	KEValue	Sample appearance
			Embodiment 1	0.271	Have no oil droplet
Embodiment 2	0.273	Have no oil droplet
			Embodiment 3	0.287	Have no oil droplet
Embodiment 4	0.292	Have no oil droplet
			Embodiment 5	0.294	Have no oil droplet

Experiment conclusion: injection stability of the present invention is good, can longer-term storage, be suitable for clinical application.

The experiment of 2 influence factor of experimental example

Sample obtained by embodiment 1-5 is subjected to illumination and high temperature experiment, detects the content in relation to substance in injection, As a result as follows:

2 light durability experimental result of table

3 high-temperature stability experimental result of table

By the illumination and high-temperature stability result of table 2 and table 3 can be seen that injection of the present invention under harsh storage condition according to Good stability can be so shown, wherein the stability result of embodiment 2 and embodiment 4 is especially prominent, has no any degradation Product.

Claims (9)

1. the derivative or its officinal salt injection of GLP-1 analog shown in a kind of formula (I), which is characterized in that every milliliter of note Penetrate the derivative or its considerable amount of officinal salt, 10mg hydroxypropyl containing GLP-1 analog shown in 20~40mg formula (I) in liquid Group-beta-cyclodextrin, 30~60mg NaCl and water for injection, using sodium hydroxide or salt acid for adjusting pH to 4~6,

His-Aib-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Glu-Glu-Ala- Ala-Lys-Glu-Phe-Ile-Ala-Trp-Leu-Val-Arg-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro- Pro-Ser-Lys

(Ⅰ)。

2. the derivative or its officinal salt injection of GLP-1 analog shown in formula (I) according to claim 1, special Sign is that the derivative or its officinal salt of GLP-1 analog shown in the formula (I) are 30mg.

3. the derivative or its officinal salt injection of GLP-1 analog shown in formula (I) according to claim 1, special Sign is that the sodium hydroxide is 5mg.

4. the derivative or its officinal salt injection of GLP-1 analog shown in formula (I) according to claim 1, special Sign is that the NaCl is 40~50mg.

5. the derivative or its officinal salt injection of GLP-1 analog shown in formula (I) according to claim 4, special Sign is that the NaCl is 45mg.

6. a kind of derivative for preparing GLP-1 analog shown in formula described in Claims 1 to 5 any one (I) or its is pharmaceutically acceptable The method of saline injection, includes the following steps:

1) derivative of GLP-1 analog shown in formula (I) or its officinal salt are dissolved with injection solvent；

2) sodium chloride and hydroxypropyl-β-cyclodextrin is added, dissolves by heating；

3) sodium hydroxide or hydrochloric acid is added, adjusting pH value is 4~6；

4) water for injection constant volume is used, is filtered, bottling.

7. the derivative of GLP-1 analog shown in preparation formula (I) according to claim 6 or its officinal salt injection Method, which is characterized in that the solvent for injection is selected from ethyl alcohol, propylene glycol, polyethylene glycol, benzyl alcohol and glycerol.

8. the derivative of GLP-1 analog shown in preparation formula (I) according to claim 6 or its officinal salt injection Method, which is characterized in that the pH value is 5.

9. according to claim 1~5 the derivative containing GLP-1 analog shown in formula (I) described in any one or its can medicine With saline injection in preparation for treating non-insulin-dependent or insulin-dependent diabetes mellitus or treating the drug of obesity In purposes.