TW492869B - Use of GIP-1 or analogs to abolish catabolic changes after surgery - Google Patents

Abstract

This invention provides a method of attenuating post-surgical catabolic changes and hormonal responses to stress. GLP-1, a GLP-1 analog, or a GLP-1 derivative, is administered at a dose effective to normalize blood glucose.

Description

492869 Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 5. Description of the invention Background of the invention 1. The scope of the invention The invention relates to an antique * β > method, which is used to prevent catabolic reactions caused by surgical trauma. M, 4, ^ t, human heart, "'Improved insulin resistance improved postoperative recovery. 2. Background Information Every year in the Western world, there are about 20-25 per million people, and there is no surgical treatment. Surgery, just like any trauma, has a marked change in all-start metabolism [Shaw, JHF, et al., Ann Surg, 2009: 63 72 (1989); Lmle, R. A., et al., PrOg Clln. Bl〇1 Res 263a: 463-475 (1987); Frayn, K.5 N. Clin. Endocrinol. 24: 577-599 (1986) 'Brandi, L., et al., Clin. Sci. 85: 525-35 (1 993)]. Accelerated synthesis of glucose, the main fuel for tissue repair, is an important metabolic change after surgery and costs protein and energy stocks in the body [Gump. FE, et al., J. Trauma, 14 ·· 378-88 (1974): Black, R · B ·, et al., Ann. Surg. 196: 420-35 (1982)]-Previously, we attributed these changes to glucose-regulated stress hormones released in response to trauma and others such as cytokines Catabolic factors. The more obvious changes in metabolic breakdown, the higher the pathogenicity, the slower the recovery of patients [Thorell, A ·, et al., Eur. J. Surg ·, 159: 593-99 (1993); Chernow, B ·, et al. ,, Arch. Intern · Med., 147 ·· 1 2 73 -8 (1 9 8 7)] 〇 Catabolic status after surgery may be treated with anabolic hormones, especially growth hormone and IGF-1 [Hammarkvist, F., etc. ·, Ann. Surg ·, 2 16 (2): 184-190 (1991); Ziegler, T ·, et al ·, Annu. Rev ·

Med., 45: 459-80 (1994): Ziegler, TR, et al. J. Parent, 4- This paper size applies to the Chinese National Standard (CNS) A4 specification (210X297 mm) (Please read the precautions on the back first ^ ^ This page) • Equipment · 492869 Α7 Β7 Printed by the Consumers' Cooperative of the Central Bureau of Standards and Economy V. Invention Description (2) Ent · Nutr · 14 (6): 5 74 _81 (1990)]. Some studies have shown significant benefits of treating insulin with catabolic trauma patients [Hinton, p., Et al., Lancet, 17 (April): 767. 69 (1971); Shizgal, H. ^. ^ Am. J. Chem. Nutr., 50: 1355-63 (1989); Woolfson, et al., N · Clin · Nuti ·, 50: i355_63 (1989); W〇lfsoon, AMJ ·, et al., N. Engl J. Med. 300: 14-17 (1979): Brooks, D., et al., J. Surg. Res. 40: 395-405 (1986); Sakurai, Y., et al., Ann. Surg. 222 : 283-97 (1995)] 0 However, there are other studies that show that the benefits of insulin on postoperative surgery are often compromised by insulin resistance. In the case of insulin resistance, normal insulin concentrations always cause sub-normal responses. Insulin resistance may be due to decreased binding of insulin to cell surface stylistics or to intracellular metabolic changes. The first type is characterized by decreased insulin sensitivity and can typically be overcome by increasing insulin concentration. The second type is characterized by a decrease in the reactivity to insulin and cannot be overcome with a large amount of insulin. f Insulin resistance after injury can be overcome by using a dose of Tengdaosu, which is proportional to the degree of insulin resistance, and the decrease in Tengdaosu sensitivity [Brandi, L S. ', etc., Chn. Science 79: 443-450 (1990); Henderson, A.A. ', et al., Chn Sci 80: 25-32 (i99〇)]. After selective abdominal surgery, the sensitivity of Tengdaosu decreased for at least five days, but not more than three weeks, and it was the most severe on the first day after surgery, and it may take three weeks to normalize [Th〇reU, A ·, Etc., (1993)]. The cause of the transient insulin resistance after trauma was not known by the two students. Corticosteroids and Glucagon: Both may be related to catabolism after trauma (please read the precautions on the back first \ ^^% this page) • Packing. Ordering

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492869 A7 B7 Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 5. Description of the invention (3) The response is related to [Alberti, KGMM, etc., J. Parent · Ent · Nutr. 4 (2): 141-46 (1980) Gelfend, RA, et al., J. Clin. Invest 74 (December issue): 2238-2248 (1984); Marliss, EB, et al. J Clin. Invest. 49: 2256-70 (1970)]. However, studies of postoperative insulin resistance in the opponent have failed to show any correlation between these catabolic hormone changes and postoperative insulin sensitivity changes [Thorell A., et al. (1993); Thorell A., Karolinska Hospital and Institute , 10401 Stockholm, Sweden (1993); Thorell A. et al., Br. J, Surg 81: 59-63 (1994)] 〇 Increased lipid utilization after trauma can blame insulin resistance via glucose-fatty acid cycle [ Randle, PJ, et al., Diab Metab. Rev. 4 (7): 623-38 (1988)]. Increased availability of free fatty acids (FFA) will flaunt insulin resistance and change the substrate oxidation from glucose to fat, even when insulin is infused at the same time [Ferrannini, E ·, et al.,]. (: 1111.11 ^ 51 72: 1737 -47 (198j) · Bevilacqua, S ·, et al., Metabolism 36: 502-6 (1987); Bevilacqua, S ·, et al., Diabetes 39: 383-89 (1990); Bonadonna, RC ·, et al · Am. J. Physiol. 259. E736-50 (1990); Bonadonna, RC, et al., Am. J. Physiol. 257: E49-56 (1989)]. Selective surgery is usually performed after overnight food. To reduce the risk of anesthesia. This established practice of fasting patients overnight (10-16 hours) before surgery can enhance the development of catabolic states and worsen insulin resistance. Studies on oppressed rats show that fasting time less than 24 hours will significantly affect the catabolic response to trauma [Alibeg〇vie -6-This paper size applies Chinese National Standard (CNS) A4 size (210X297 mm) (k first Read the back page page}. Binding · Order 492869 A7 B7 V. Invention (4 Printed by A., etc. of the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs, Circ. Shock, 39 .. 1-6 (1993); Eshali, A. H., etc .. Eur. J. Surg., 157: 85-89 (1991); Ljungqvist, 〇, et al. Am. J. Physiol., 22: E692-98 (1990)]. Even fasting for a short time before trauma in rats will significantly reduce sugar storage, deeply Change hormone environment, increase stress response and most importantly increase mortality [Alibegovie, A., et al., Circ. Shock, 39: 1-6 (1993)]. Glucose is administered before surgery, regardless of oral administration [Nygren, J., et al., Ann Surg. 222..728-34 (1995)] or reduced insulin resistance after surgery compared with fasting patients. Overnight glucose infusion (5 mg / Kg / min) patients lost an average of 32% of their insulin sensitivity after surgery, but patients who underwent routine overnight fasting lost an average of 55 ° / o [Ljungqvist, 0, etc.] Coll. Surg. 178: 329-36 (1994)] 〇 In addition to the adverse effects of fasting on postoperative recovery, the patient is undergoing surgery And thereafter the inactive and low-calorie nutrition also increase insulin resistance after the surgery. For healthy subjects, inactivity and low-calorie nutrition for 24 hours showed that peripheral insulin resistance could be increased by 20-30% in healthy volunteers. Therefore, the post-operative pancreatic [Ljungc ^ st, 〇., (1994)] reported by preoperative glucose infusion may be partly due to the addictive effect of post-operative beds: interest and low-calorie nutrition. In view of the prevalence of surgery, it is important to minimize negative side effects, such as catabolic reactions and insulin resistance, in order to improve recovery and reduce mortality. Insulin resistance after surgery enables the use of tossin to treat catabolic conditions ~ Treatment Text frustration. Determined fasting practices before surgery Aggravate differences after surgery

(Please read the back, v precautions 舄 this page first) • Binding line 492869 A7 Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 5. Description of invention (5 states and insulin resistance. Therefore, the need can be overcome at the same time Catabolism and insulin resistance treatment., As disclosed herein, 'can overcome this catabolic situation and insulin resistance (the treatment is administered before surgery, from time to time and after administration of glucose and insulin. However, insulin The possibility of hypoglycemia caused by perfusion. Hypoglycemia is defined as the concentration of blood glucose below 0.3 millimolar. Hypoglycemia increases the risk of arrhythmia and is a dangerous result of insulin perfusion. To prevent hypoglycemia people have developed diabetes perfusion insulin rules System [Hendra, Ding Yi, et al., Diabetes Res. Clin. Pract, 16: 213 22〇 (my]. However, under this rule system, 21% of patients still have hypoglycemia. Glucoside after another myocardial infarction_ In the glucose control study, hypoglycemia occurred in 18% of patients when perfused with tolin and glucose [Malmberg, κ A, et al.,

Diabetes Care, 17 · 1007-1014 (1994)]. Insulin administration > Wang Shi also needs to constantly monitor blood glucose levels so that hypoglycemia can be detected and treated as soon as possible. In the cited study, those who received Tengdaosu perfusion [Ma 丨 mberg, 1994] measured blood glucose at least every two hours and adjusted the perfusion rate accordingly. Therefore, the safety and efficiency of insulin-glucose infusion therapy for patients with myocardial infarction depends on the convenience and rapid availability of blood glucose data. This strong need for blood glucose monitoring has put pressure on medical staff and increased treatment = prescription and cost. As a result, the clinical care unit before the operation is usually not the same as that in the intravenous vein; the Wangshe insulin provides monitoring and optimization of blood glucose before the operation. Considering the dangers and burdens of the nature of insulin perfusion requires alternative anterior / posterior-surgical approaches to the degrading metabolic response to trauma. Insulin-stimulating insulin, a glucagon-like peptide}, abbreviated as (} Eb 1, the size of this paper is applicable to the Chinese National Standard (CNS) A4 specification (210X297 mm) (please first read the precautions on the back of the HP written version) (Page)-Outfit.-Ding 492869 A7

Glucagon plus 4 before the intestine is used to obtain and enhance nutrients. Induced insulin release [kymann B., et al., Lancet 2... 13] [03] (1987)]. The same type of truncated G L P 1 can stimulate the secretion of insulin (insulin-promoting effect) and CAMP form [see, for example, Mojsov s int;% other

Protem Research, 40: 333_343 (1992)]. Various different in vitro experiments and mammals have been established, especially humans—the relationship between the administration of exogenous ⑽ · 1, GLP-U7-36) amine and GLp] (7-37) acid-stimulating insulin response [for example, see Nauck, M.A., et al., Diabet 〇〇〇gia,%: 741-744 (1993); Gutniak, M, et al., New England] 〇f

Medicine, 326 (20): 13 16-1322 (1992); Nauck, MA, et al., J. Clin. Invest., 91: 301-307 (1993); and Thorens, B., et al., Diabetes, 42. 1219-1225 (1993)]. By stimulating tannin sensitivity and intensifying glucose-releasing insulin release at physiological concentrations, ^! ^! ^ 1 (7-3 6) amine test produces a significant anti-diabetic effect on patients with insulin-dependent diabetes [Gutniak M., et al., New England J. Med. 326: 1316-1322 (1992)]. When administered to non-insulin-dependent diabetic patients, GLP-1 (7-3 6) amidine stimulates insulin release, reduces glycocrin secretion, inhibits gastric emptying and enhances glucose utilization [Nauck. 1993; Gutniak, 1992; Nauck , 1993] 0 Long-term therapy with GLP-1 molecules is hindered by the short half-life in serum of GLP-1 peptides. For example, G L P-1 (7-3 7) has a serum half-life of 3 to 5 minutes. GLP-l (7-36) amidamine has a half-life of about 50 minutes when administered subcutaneously. Therefore, in order to achieve long-term effects, these molecules must be continuously infused [Gutniak M., etc., Diabetes Care 17: 103 9-1044 -9- This paper size applies to the Chinese National Standard (CNS) A4 specification (210X297 mm) (Please read the historical notes on the back of Sr first-r this page) Order Printed by the Consumer Standards Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 492869

V. Description of the invention Printed by the Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs (1994)]. In the present invention, the short half-life of GLp_i and therefore the need for continuous administration are not disadvantages, as patients are typically admitted to the hospital before surgery and fluid is continuously parenterally administered during and after surgery. SUMMARY OF THE INVENTION The present invention therefore presents, for the first time, a method for attenuating catabolic changes and insulin resistance after surgery, which comprises administering to a patient in need selected from the group consisting of GLpq, GLP-1 analogs, GLP-1 derivatives and pharmaceutically acceptable salts thereof . The figure is briefly explained. Figure 1 shows the postoperative (p0st〇p) control period of 7 patients () and 6 control patients (0) who received hyperinsulinemia and normal blood glucose perfusion before and during selective surgery. Graph of percentage change in glucose perfusion rate (GIR) in the pre (preop) control period. Figure 2 is a graph showing the effect of continuous G L P-1 (7-3 6) ammonium perfusion on mean blood glucose concentration (¾mol) (5) in 5 patients with non-insulin-dependent diabetes mellitus (NIDDM) at night. The figure also shows the effect of continuous insulin perfusion on the average blood glucose concentration of the same MAniddm patient on different nights (0--). Figure 3 is a graph showing the effect of GLP-1 (7-3 6) hydrazine perfusion on the mean blood glucose concentration (millimolars) (___) of 5 NIDDM patients during the day before the start of three meals per meal. The graph also shows the effect of subcutaneous injection of insulin on the mean blood glucose concentration (--0--) of the same 5 NIDDM patients on the other day using injections shortly before each meal. Detailed description of the invention G L P-1 '' means G L P-1 (7-3 7). According to the habit of this technique, _____________ ~ 10-This paper size is applicable to China National Standard (CNS) 8-4 specifications (210X297 meals) (Please read the back • notice beforehand ▲ • Installation-Shu write this page) Thread • is 1— 1— 111 mm ϋϋ 492869 A7 printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs _________B7___ V. Description of the invention (8) GLP-1 (7-37) amine end has been named No. 7, and the carboxyl group The end number is 3 7. The amino acid sequence of G L P-1 (7-3 7) is well known in the art, but is listed in the following table for the convenience of the reader:

NH2-His7-Ala-Glu-Gly10-Thr-Phe-Thr-Ser-Asp1: > -Val-Ser-Ser-Tyr-Leu20-Glu-Gly-Gln-Ala-Ala25-Lys-Glu.Phe-Ile -Ala30-Trp-Leu-Val-Lys-Gly3) -Arg-Gly37-COOH (SEQ ID NO: 1) "GLP-1 analog" is defined as having one or more amino acid substitutions compared to GLP-1, Molecules for deletion, inversion or addition. GLP-1 analogs known in the art include, for example, g LP-1 (7-3 4) and GLP-1 (7 -35), GLP-1 (7-36), Gln9-GLP-1 (7-37 ), D-Gln9-GLP-1 (7-37), Thr16-Lys18-GLP-1 (7-37), and Lys18-GLP-1 (7-37). Preferred GLP-1 analogs are GLP-1 (7-34) and GLP-1 (7-3 5), which are disclosed in U.S. Patent No. 5,1 18,666, which is incorporated herein by reference and also includes GLP-1 ( 7-3 6), which is a bioprocessed version of GLP-1 with insulinotropic properties. Other GLP-1 analogs are disclosed in U.S. Patent No. 5,545,618 and are incorporated herein by reference. "GLP-1 derivative" is defined as having an amino acid sequence of GLP-1 or GLP-1 analogues, but having one or more amino acid branches,-carbon atoms, terminal amine groups or terminal carboxylic acid groups Chemically modified molecules. Chemical modifications include, but are not limited to, adding chemical moieties, creating new bonds, and removing chemical moieties. Located at -11-This paper size applies the Chinese National Standard (CNS) Α4 specification (210 × 297 mm) 'I --------- see-(please read the notes on the back and write this page first) Order Line 492869 Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs A7 B7 V. Description of the invention (9) Modification of amino acid branching chain includes' but not limited to, brewing of ε-amino group of lysine, arginine, group N-firing of lysine or lysine, re-firing of weil acid or aspartic acid and de-amination of n-amine or aspartic acid. Modifications of the terminal amine group include, but are not limited to, deaminyl group, N-low post-burning group, ν-di-low-carbon fe group and N-SS: group modification. Modifications of the terminal groups include, but are not limited to, amidoamine, alkanoylamine: amine, dialkyl amine, and oligocarbonate ester modification. Lower alkyl is C! -C4 alkyl. In addition, one or more branched or terminal groups can be protected with protecting groups known in general protein chemistry. The α-carbon of the amino acid may be mono- or dimethylated. A preferred class of GL P-1 analogs and derivatives used in the present invention are composed of molecules of the following formula: Ri-X-Glu-Gly10-. Thr-Phe-Thr-Ser-Asp15-Val-Ser-Ser. Tyr-Leu20 «Y -Gly-Gln.Ala-Ala25-Lys« Z -Phe-Ile-Ala30. Trp-Leu-Val-Lys-Gly35-Arg-R2 (SEQ ID NO: 2) and its pharmaceutically acceptable Salt, wherein: R1 is selected from the group consisting of histidine, D-histamine parent, deamino-histamine § parent, 2-aminohistamine, y-light histamine, homohistamine, α_fluorofluorenylhistamine and α_fluorenylhistamine; x is selected from Ala, Gly, Val, Thr, lie and han-fluorenyl-Ala; γ is selected from Glu, Gin, Ala, Thr, Ser and Gly; Z is selected from Giu, Gin, Ala, Thr, Ser, and Gly; and R2 is selected from NH2 and Gly-OH; its restrictions are ___ 12-_ _ This paper size is suitable for financial standards CNS) A4 specification (2! GX297 public meal) " ----- ^ (Please read the precautions on the back ^^ [page])-binding * 492869 A7 B7 Staff Consumer Cooperatives, Central Standards Bureau, Ministry of Economic Affairs 5. The description of the invention (10) is that the isoelectric point range of the compound is from about 60 to about 9.0, and it is further < The limitation is that when R ^ His, x is Ala, Y is Glu, and z is Glu, then 112 must be nh2. Many GLP- 丨 analogs and derivatives with isoelectric points in this range have been shown and include, for example: GLP-1 (7-36) NH2 Gly8-GLP-1 (7-36) NH2 Gln9-GLP-1 (7-37) D-Gln9-GLP-1 (7-37) acetyl-Ly s9-GLP-1 (7-37) Thr9-GLP-1 (7-37) D-Thr9-GLP-1 (7- 37) Asn9-GLP-1 (7-37) D-Asn9-GLP-1 (7-37) Ser22-Arg23-Arg24-Gln26-GLP-1 (7-3 7) Thr16-Lys18-GLP-1 (7 -37) Lys18-GLP-1 (7-37) Arg23-GLP-1 (7-37) Arg24-GLP-1 (7-37), and similar [Ref., For example, x, W91 / 11457] 〇 Another preferred type of active compound used in the present invention is +, λ ^ W〇91 / 1 1457 and actually includes GLP-1 (7-34), GLP-1 (7_, < v ° 5), ____ -13- _ I Paper size applies Chinese National Standard (CNS) A4 specification (21 × 297 mm) '' ~~ ^ ----- (Please read the precautions on the back first v -install-f this page }, 11 line 492869 Α7 B7 V. Description of the invention (11) GLP-1 (7-36) or GLP-1 (7-37) or its amide type and its pharmaceutically acceptable salt, at least one selected from the group consisting of The following modifications: (a) The lysine at position 26 and / or 34 is replaced by the following: glycine, serine, cysteine Acid, threonine, asparagine, glutamine, tyrosine, alanine, valine, isoleucine, leucine, methionine, phenylalanine, arginine, or D- Lysine, or the 36-position arginine is replaced by glycine, serine, cysteine, threonine, asparagine, glutamine, tyrosine, alanine, Valine, isoleucine, leucine, methionine, phenylalanine, lysine or D-spermine; (b) Tryptophan at position 31 is oxidatively resistant amino acid Substitution; (c) at least one of the following substitutions: replacement of valine at position 16 with tyrosine, serine at position 18 with aspartic acid, and glutamic acid at position 21 with aspartic acid, Serine replaces glycine at position 22, arginine replaces glutamine at position 23, alanine at position 24, alanine at position 24, and lysine at position 2 6 with glutamine, and ( d) at least one of the following substitutions: glycine, serine, or cysteine to replace alanine at position 8, aspartic acid, glycine, serine, cysteine, threonine, Asparagus Amines, lysosamines, tyrosines, alanines, vals, amines, isoleucines, leucines, thiosamines, or phenylalanines replace position 9 glutamate, serine, cysteine Glycine, threonine, asparagine, glutamine, tyrosine, alanine, valine, isoleucine, leucine, methionine or phenylalanine at position 10 Glycine and glutamic acid replace aspartic acid at position 15: and (e) Glycine, serine, cysteine, threonine, asparagine, -14- This paper applies to China National Standard (CNS) Α4 ^ (210X297 mm) (Please read the v notes on the back first \ 1 ||| ^ this page) Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 492869 A7 B7 V. Description of the Invention (12 ) Wemiamine, lysine, alanine, valine, isoleucine, leucine, thiothiamine, or phenylalanine or D- or N-halide or alkylated histidine substitution positions When the substitution of histamine '7' in (a), (b), (d) and (e) is performed, the amino acid for substitution may be D-type as the case may be, and the amino acid for substitution at position 7 may be substituted. Depending on the situation It is N-halide or N-alkylation type. Because of the enzyme, dipeptidyl-peptidase IV (DPP IV) may be related to the rapid inactivation of GLP-1 administered in our eyes [see, for example, Men 11 ein, R., et al., Eur. J · Biochem., 2 14: 829-835 (1993)], administration of GLP-1 analogues and derivatives that prevent DPP IV action is preferred, administration of Gly8-GLP-1 (7-36) NH2, Val8-GLP-1 (7-37) OH, a-fluorenyl-Ala8-GLP-1 (7-36) NH2 and Gly8-Gln21-GLP-1 (7-3 7) 〇Η or a pharmaceutically acceptable salt thereof are more preferable. If the present invention uses U.S. Patent Nos. 5,8,8,6,6, it is better to apply for a molecule, which is clearly incorporated herein by reference. The molecule is selected from peptides with the following amino acid molecules: NH2-His7-Ala.Glu-Gly10-

Thr-Phe-Thr-Ser-Asp15-Val-Ser-Ser-Tyr-Leu20-

Glu-Gly-Gln-Ala-Ala25-Lys-Glu-Phe-Ile-Ala30-

Trp-Leu-Val-X (SEQ ID NO ·· 3) where X is selected from Ly s and Ly s-Gly, and derivatives of the peptide, wherein the peptide is from: a pharmaceutically acceptable acid of the peptide Into a salt, the medicine of this peptide can be accessed _____- 15- This paper size is applicable to China National Standard (CNS) A4 specification (2i0x297 mm)

(Please read the “Notes on the back side of the book I * install-π write this page first”) Print the A7 B7 printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs i. Description of the invention A pharmaceutically acceptable amidine that accepts a lower alkyl ester and the peptide selected from the group consisting of amidine, lower alkyl amidoamine and lower alkyl dialkyl amidoamine. Another preferred compound used in the present invention includes U.S. Patent No. 5,5 12,549, which is clearly incorporated herein by reference and has the following general formula: R ^ Ala-Glu-Gly10-

Thr-Phe-Thr-Ser-Asp1: > -Val-Ser-Ser-Tyr-Leu20-

Glu-Gly-Gln-Ala-Ala25-Xaa-Glu-Phe-Ile-Ala30-

Trp-Leu-Val-Lys-Gly35-Arg-R3

I R2 (SEQ ID NO: 4) and a pharmaceutically acceptable salt thereof, wherein R 1 is selected from the group consisting of 4-imidazolidine, 4-imidazolium or 4-imidazol-αα-diamidylacetamyl R2 is selected from c6_c10 unbranched fluorenyl groups or is absent, R3 is selected from Gly-OH or NH2, and Xaa is Lys or Arg, which can be used in the present invention. The more preferred compound of SEQ ID NO: 4 used in the present invention is one in which the parent "is Arg and R2 is a C6-Ci0 unbranched g helmet group. The particularly preferred compound of SEQ ID NO: 4 used in the present invention is one of xaa Is Arg and R2 is a C6-C1 () unbranched fluorenyl group and R3 is Gly-OH. The compounds of SEQ ID NO: 4 which are good for use in the present invention are those in which xaa is Arg and R2 is C6-C1 () Non-branched fluorenyl group, which is Giy_〇H and Ri is 4_Mi Jun propionyl group. -16- This paper size applies to China National Standard (CNS) A4 specification (210X297 mm) I ----- ---- 1-- (please read the notes on the back V for the first copy), and order it printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 492869 printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs A7 B7 14) The best SEQ ID NO: 4 compounds used in the present invention are those in which Xaa is Arg 'R is 0 8 unbranched fluorenyl group, r3 is Gly-OH, and R1 is 4-methylimidinopropyl group. The present invention The molecule patented in US Patent No. 5320, 7 12 is particularly preferred and is clearly incorporated herein by reference. The molecule is selected from peptides having the following amino acid sequence: NH2-His7-Ala-Glu-Gly10-Th r-Phe-Thr-Ser-Asp15-Val-Ser-Ser-Tyr-Leu20- Glu-Gly-Gln-Ala-Ala25-Lys-Glu-Phe-Ile-Ala30- Trp-Leu-Val-Lys-Gly35- Arg-Gly37-COOH (SEQ ID NO ·· 1) and a derivative of the peptide, wherein the peptide is selected from the group consisting of a pharmaceutically acceptable acid addition salt of the peptide, a pharmaceutically acceptable carboxylic acid salt of the peptide, the peptide The pharmaceutically acceptable low-carbon alkyl ester and the peptide are selected from the group consisting of amidine, low-carbon alkylamidine, and low-carbon dialkylamidine. The present invention uses GLP-1 (7- 36) Amidoamine or its pharmaceutically acceptable salt is the best. The amino acid sequence of GLP-1 (7 · 3 6) amidoamine is: NH2-His7-Ala-Glu-Gly10-Thr-Phe-Thr-Ser- Asp15-Val-Ser-Ser-Tyr-Leu20-Glu-Gly-Gln-Ala-Ala2: > -Lys-Glu-Phe-Ile-Ala30-Trp-Leu-Val-Lys-Gly3) -Arg-NH2 ( SEQ ID NO: 5) _-17-_ This paper size applies the Chinese National Standard (CNS) A4 specification (21 OX 297 mm) (please read the V notice on the back page first page) • Binding · Order 492869 Economy Printed by the Consumer Standards Cooperative of the Ministry of Standards of the People's Republic of China A7 _______B7 V. Description of the Invention (15) '' The active compound used in the present invention, that is, GL p _ i, GL p _ 丨 is similar Or GLP-1 bio-organic preparation methods are well known and described in U.S. Patent Nos. 5,1 18,666, 5,120,712 and 5,523,549, which are incorporated herein by reference. 2. The amino acid portion of the active compound used in the present invention or its precursor is borrowed from the company) Solid-phase synthetic chemistry, 2) Purified from natural sources GLp molecular formula 3) Recombination] 31 ^ 8 technology. Solid-phase chemical synthesis is well known in the art and may be found in general internal tables such as the following: 'Dugas, H. and Penney, C., Bioorganic Chemistry, Springer-Verlag, New York (1981), pages 54-92, Mernfield JM, Chem. Soc., 85: 2149 (1962), and Stewart and Young, Solid Phase Peptide Synthesis, Freeman, San Francisco (1969) pp. 24-66. For example, the amino acid portion can be synthesized by a solid-phase method using a 43A peptide synthesizer (PE-Applied Biosystems, Inc., 850 Lincoln Center Drive, Foster City, CA 94404) and a synthesis cycle provided by PE-Applied Bio systems. . B O C -amino acids and other reagents are available from PE-Applied Biosystems and other chemical suppliers. We applied the sequence Boc chemistry using the dual coupling method to the starting p-phenylenediphenylphenylamine resin for the production of C-terminal carboxamide. For the production of C-terminal acids, we use the corresponding PAM resin. A s η, G1 η and A r g are coupled using a preformed light phenyl hydrazone triazole ester. May use the following branched protective groups: .Arg, tosylsulfonyl Asp, cyclohexyl. __- 18 -___ This paper size applies to China National Standard (CNS) A4 (210X297 mm) ------ ----- 11 — (please read the 5 items on the back page and write this page first) Order A7 printed by the Consumers' Cooperatives of the Central Bureau of the Ministry of Economic Affairs ------------ B7 V. Description of the invention (16) G1 u, cyclohexyl Ser, fluorenyl T hr, benzyl Tyr, 4-bromocarbonylfluorenyloxy B oc can only be deprotected using trifluoroacetic acid in methane solution. After synthesis, The peptide can be deprotected and the resin can be cut off with anhydrous hydrofluoric acid (HF) containing 10% m-phenol. The removal of the branched protecting group and the peptide from the resin is performed in 5 C to 5 C to 60 minutes in ice is preferred. Remove, wash the peptide / resin with ether, and the peptide is extracted with glacial acetic acid and freeze-dried. Recombinant DNA technology well known to those skilled in the art can also be used to prepare the active compounds used in the present invention. In fact, it may be better because of the higher yield of the recombinant method. The basic steps of the recombinant production method are: a) Isolation of the natural D n A sequence encoding the GLP-1 molecule Or construct a synthetic or semi-synthetic molecule encoding GLP-1 0να, b) place the coding sequence on a expression vector in a manner suitable for expression alone or as a fusion protein, c) transform the expression vector into a suitable triton or Prokaryotic host cells, d) culturing the transformed host cells under conditions that allow the expression of the Olp-1 molecule, and e) recovering and purifying the recombinantly produced glP-1 molecule. As mentioned above, the coding sequence must be a fully synthesized or larger modified product of the primary glycated DNA. In Lund et al., Proc Natl. Acad. Sci USA 345-349 (1982), a D Ν Α sequence encoding a pre-glucagon can be seen and the desired result can be obtained by changing the nascent sequence as a hemi-combination (please first w Notes on the back of the reading * ml copy book education)-installed ·, ιτ 线 -19- 492869 Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs and Ministry of Education 5. Description of the invention (17 Starting materials for the compounds of the present invention. Click. People know to use this Known methods of constructing in vitro or in vivo transcription and TT # can cause synthetic genes produced by the GLP-1 molecule. Technicians who understand that the genetic code can be constructed to encode all GLP- 1 Large number of molecules but a fixed number of Dn A sequences. Methods for the construction of synthetic genes are well known to those skilled in the art. See Brown, et al. (1979) Methods in Enzymology, Academic Press, NY, 68% 1 5 1. DN A sequence system The genetic code is designed by the required amino acid sequence, and it is easy for ordinary biologists to recognize the genetic code. Once sighed, the sequence itself must use traditional DNA synthesis equipment such as 380A or 380B DNA Synthesizer (Manufactured by PE-APPlled Biosystems, Inc., 850 Lincoln Center Drive, Foster City, CA 94404). In order to express the amino acid portion of the compound used in the present invention, we will synthesize DNA by appropriately restricting the use of endonucleases The sequence is inserted into any of a number of suitable recombinant DNA expression vectors. Refer to General Maniatis et al. (1989) Molecular Cloning; A Laboratory Manual, Cold Springs Harbor Laboratory Press, NY ·, 1-3. We are encoding the GLP-1 molecule. One end of the DNA is designed to limit the cutting position of the endonuclease to facilitate its amplification and separation and integration of expression vectors that are well known in the art. The specific endonuclease used is controlled by the restriction endonuclease cutting pattern of the parental expression vector used. I choose a restricted position that can appropriately point to the coding sequence with the control sequence, so that I can read and express the desired protein properly in-frame. I should locate the coding sequence in the promoter and the ribosome of the expression vector. The proper reading frame of the binding position, the latter two are the host of the desired protein-20 This paper size applies Chinese National Standard (CNS) A4 regulations Grid (210X297 mm)

(Please read "Precautions on the back of I"-> Write this page first) Thread 492869 A7 B7 Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 5. Description of the invention (18) The cell has functions. For efficient translation of synthetic genes, they need to be operably linked to the promoter-operon region. Thus, the promoter-operon region of the synthetic gene is placed in the same sequence direction as the A T G start codon of the synthetic gene. A variety of expression vectors that can be used to transform prokaryotes and prion cells are well known in the art. See The Promega Biological Research Products Catalogue (1992) (Promega corp ·, 2800 Woods Hollow Road, Madison, WI, 5371 1-5399); and The Stratagene Cloning Systems Catalogue (1992) (Stratagene Corp., 11011 North Torrey Pines Road, La Jolla, CA, 92037) plus U.S. Patent No. 4,710,473 indicates that a circular O1 ^ octaplasma transforming vector that can express high amounts of foreign genes in E. coli can be used. These plastids can be used as transforming vectors in recombinant DNA methods, and a) give the plastids the ability to autonomously replicate in host cells; b) control the autonomous plastids to replicate at a temperature that maintains host cell culture; c) The maintenance of stable plastids in the host cell community; d) guides the synthesis of protein products that symbolize the plastids maintained in the host cell community; e) provides a unique array of restriction endonuclease recognition sites for plastids; and f) terminates mRNA transcription . These circular DNA plastids can be used as vectors for recombinant DNA methods in order to obtain the expression of a high amount of foreign genes. -The performance of the amino acid portion of the compound used in the construction of the present invention is shown in this paper ruler_Jiaxian ((Please read the note of Back® first-Install --- Write this page) 492869 A7 B7 Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs. 5. Description of the invention (19) 'The next step is to put the vector into a suitable cell and use it to construct a recombinant host cell that expresses the use of peptides. The cells are transformed with recombinant DNA vectors. The technology is well-known in this technology, and can also be found in, for example, Maniatis, etc., above the general literature. The host cells must be constructed from nucleus or prokaryotic cells. Prokaryotic host cells usually produce higher quality proteins than Easy to cultivate. Proteins expressed in high-volume protein expression systems have the characteristics of agglomerating into particles or inclusion bodies, and the inclusion bodies will contain a high amount of over-expressed proteins. The protein aggregates should typically be used in the art Method recovery, dissolution, denaturation and rewinding. Refer to Kreuger, et al. (199〇) h

Protein Folding, edited by Gierasch and King, pages i36-142,

American Association for the Advancement of Science Announcement Numbers 89-18S, Washington, D.C. and U.S. Patent Number 4,923,967. Some people also use well-known methods to change the amino acid sequence of GLP-1 precursor or g LP-1 analog to produce the required GLP-1 analog or GLP-1 derivative: including chemical modification of GLP-1 precursor, enzyme Modifications or a combination of chemical and enzymatic modifications. The GLP-1 molecules used in the present invention can also be produced by classical solution-phase methods and semi-synthetic methods. Methods for preparing the GLP-1 molecules of the present invention are well known to those skilled in peptide chemists. The fluorenyl group may be added using any of a variety of methods well known in the art; the epsilon-amine group of Lys. See, Bioconjugate Chem. &Quot; Chemical Modifications of Proteins: History and Applications, pp. 1, 2-12 (199〇) and Hashimoto et al., Pharmaceutical Res. 6 (2): 171-176 (1989 -22- This paper size is in accordance with Chinese National Standard (CNS) A4 specifications (21 〇X 297 male sauce) (Please read the precautions on the back β to write this page). Binding and ordering 492869 A7 B7 Printed by the Staff Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs V. Description of the invention (20) For example, the use of 50% acetonitrile in boric acid buffer solution can add the "hydroxy_ tiger ί subline" of octanoic acid to the amine group -ε amine. The skin can be added to the taste bite group < Alcoholization can be performed later. Furthermore, if the peptide is made by recombinant methods, it may be hydrolyzed before enzymatic cleavage. Furthermore, as described in WO 96-29342, the lysine production of GLP-1 bamboo organisms is feasible. The functions of various protected, unprotected and partially protected natural, unnatural GLP-1 (7-36) amidamine and GLP-1 (7-37) molecules have been explained in this art. Analogs and Derivatives [See, for example, U.S. Patent Nos. 5,120,7 12 and 5,1 18,666, which are incorporated herein by reference. Photo, with Orskov, C., et al., J. Biol. Chem., 264 (22) · 12826-12829 (1989) and WO 91/1145 7 (Buckley, DI, et al., August 1991 8th publication). The amine and carboxy-terminated amino acid residues of GLP-1 derivatives can be protected as appropriate, or only one end can be protected. The formation and removal reactions of this protecting group are described in "quasi-operations" Includes, for example, "Protective Groups in Organic ChemistryM, Plenum Press, London and New York (19 73) Green, TH" " Protective Groups in Organic Synthesis " Wiley, New York (1981); and The Peptides ,,, Volume 1 Schr & amp der and Lilbke, Academic Press London and New York (1965). Representative amine protecting groups include, for example, methylamido, ethylamido, isopropyl, butoxycarbonyl, carbamoylcarbonyl, carbonyloxy And similar. Representative carboxy protecting groups include, for example, benzyl ester, methyl ester, ethyl ester, tert-butyl ester, p-nitrophenyl ester and the like. The compound base used in the present invention, low post-burning S purpose, GLP-1 derivatives are based on the desired (CrC4) alkanol and the desired peptide under the conditions of a catalytic acid such as hydrochloric acid. 23 (Please read the back_ Precautions ^ .Installation-F this line thousand T " a {a public 7 9 2 492869 A7 B7 printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 5. Invention Description (21 anti-iC 'into 4 k base S Suitable conditions for the formation include a reaction temperature of about 5 ° C and a reaction time of about} hours to about 3 hours. Similarly, alkyl ester derivatives of a 邛 and / or Glu residues can also be formed. Compounds used in the present invention Carboxamide derivatives are prepared as described in, for example, Stewart, JMSolid Phase Peptide Synthesis, Pierce Chemical Company Press, 1984. GLP-1, GLP- 丨 analogs or GLpq derivatives can be used in the present invention. Pharmaceutically acceptable salts. The acids commonly used to form acid addition salts are inorganic parent materials such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid, and the like and organics such as p-toluene acid.曱 续 续 酸, oxalic acid, p-bromobenzilin, gray § & Citric acid, citric acid, acetic acid and the like. Examples of such salts include sulfate, pyrosulfate, double sulfate, sulfite, double sulfite, phosphate, monohydrogen phosphate, dihydrogen phosphate Salt, metaphosphate, pyrophosphate, chloride, bromide, salt, iodide, acetate, propionate, caprate, caprylate, acrylate, formate, isobutyrate, Acid salt, heptanoate salt, propargyl acid salt, oxalate salt, malonate salt, calamate salt, suberate salt, sebacate salt, fumarate salt, maleic acid salt, Butyl-1,4-diacid, hexadecyl-i, 6-diacid, dibasic acid salt, pirate, fluorenyl phosphonate, dinitrobenzate, hydroxyhenate, hydrazone Oxybenzoic acid salt, phthalic acid salt, sulfonate, xylene citric acid salt, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, r- Butyrate, gluconate, tartrate, formamate, propionate, a-1-acetate, bodamine-2, amylate and the like. Better Acid addition salt Such as hydrogen acid and hydrogen _____ 24 · This paper size is applicable to the Chinese National Standard (CNS) A4 specification (210X297 mm) '~ (please * read the notes on the back and write to this page first}. 11 492869 A7 B7 Printed by the Consumer Cooperatives of the Central Bureau of Standards, Ministry of Economic Affairs. 5. Description of the invention (22) Bromic acid, especially hydrogen acid. Test addition salts include those derived from inorganic testers, such as ammonium or gold or soil tests. Metal hydroxides, carbonates, bicarbonates, and the like. Therefore, the bases which can be used to prepare the salts of the present invention include sodium hydroxide, potassium hydroxide, ammonium hydroxide, potassium carbonate and the like. Salt type is especially good. The GLP-1, GLP-1 analog or GLP-1 derivative used in the present invention can be formulated with one or more excipients before being used in the present invention. For example, the active compound used in the present invention can form a complex with a divalent metal cation by a well-known method. The metal cation includes, for example, Zn ++, Mn ++, Fe ++, Co ++, CcT ~, Ni ++, and the like. Optionally, the active compound used in the present invention can be mixed with a pharmaceutically acceptable buffer and adjusted to the necessary stability and a degree suitable for parenteral administration. Optionally, one or more pharmaceutically acceptable antimicrobial agents may be added. The preferred pharmaceutically acceptable anti-biological agents are the meta-methanine and phenols. One or more pharmaceutically acceptable salts can also be added to adjust the ion concentration or tonicity. One or more excipients may be further added to further adjust the tonicity of the prescription. Examples of glycerin-based isotonic adjustment excipients. Application can be by any effective method known to a physician of ordinary skill. Parenteral administration is by injection into the body by a sterile syringe or some other mechanical device such as an infusion pump, which is commonly seen in the medical literature. Parenteral routes include intravenous, intramuscular, subcutaneous, intraperitoneal, intraspinal, intramembranous, intraventricular, intraarterial, subarachnoid and epidural. The intravenous, intramuscular and subcutaneous routes of administration of the compounds of the present invention are more comparable. The method of administering the compound of the present invention by intravenous and subcutaneous routes is advantageous. Right and wrong _______- 25 ^ -_ National standard of this paper (&) 8 4 specifications (21〇 '乂 297 mm)'-(Please read the precautions on the back i. Packing-1 (Write this page) Order line 492869 Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 5. Description of the invention (23, for enteric administration, the active compound of the present invention is preferably a basic distillation with an appropriate P Η Mix with water. ~ Know that other medicines can be used to control the sustained effect. Preparations for controlled release are known to be made of the active compound used in the present invention to form a complex or absorb it using a permanganate. Outside μ. ≪ Extended duration By selecting appropriate macromolecules, for example, "Amino Acid, Polyacetate Biha Bite Concentrate, Ethyl Acetate, Ethyl Acetate, Methyl Ester, Methionine, Methyl Cellulose, or Protamine Sulfate , And by selecting giant knives; Chen degree and combined methods to prolong the release. In addition, a possible method to extend the duration of the effect by using a k-release formulation is to combine the biochemical mouth and mouth of the present invention into polyester, Polyamino acid, hydrated gum, poly (lactic acid) or ethylene glycol Granules of polymer materials such as acetoacetate copolymers. Instead of incorporating chemical compounds into these polymer particles, it is possible to encapsulate the present invention in a microcapsule with a compound, the preparation of which is, for example, by the use of rubber particles Technology or interface U methods such as fluorenyl methyl cellulose or gelatin · microcapsules respectively: or using colloidal drug delivery systems such as liposomes, albumin microspheres, microemulsions, nanoparticle and nanocapsules or It is a giant emulsion. These experiences are disclosed in Remington, sphamaceuticai (1980). S According to the experience of the present invention, patients who need the compound used in the present invention receive the compound (the recipient is about 16 hours before surgery). No more than 5 days after surgery. Τ & The starting time of the compound administered by the mother before surgery is about 16 hours to 1 hour before surgery. The compound of the invention should be applied before surgery to reduce catabolic effect and insulin resistance. The length of time depends on the general technical paper is also applicable to the Chinese National Standard (CNS) A4 specifications (2ι〇χ297 public funding) (please first read the V notes on the back ^ • installed-I write (This page) Order

Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs: == What is the effect? Decision, which includes the most important ones: hard dog ^ w; or, supply of glucose infusion or drink, or-some aspects: the preparation period of the operation, and regardless of the patient's sex, weight and age to regulate the severity of blood sugar, Any rate that cannot adjust the blood sugar ratio: 2: the severity of the trauma caused, the route of administration and the bioavailability of the compound used. The best time to start-Jiashi & is cut by hand from 1 to 10 hours. She started with < Defective Period, 2 to 8 hours before surgery. Island interpreter, pancreas s ^ 'after specific type of surgery, selective abdominal surgery, is the most severe on the first day after surgery, lasts at least 5 days, and may be normalized for three weeks [Th⑽u, A, etc., ( 1993)]. Therefore, surgery 1 may require administration of the compound used in the present invention for a period of time, depending on factors known and determined by a general physician. These factors include whether the patient fasts or supplies glucose infusions or drinks or some other form of survival after surgery and is not limited to the patient's gender, weight and age, any inability to regulate blood glucose severity, any cause of inability to regulate blood glucose, trauma caused by surgery The severity is positive, the route of administration and bioavailability, the persistence in the body, the formulation, and the efficacy of the administered compound. The preferred period for administering the compound used in the present invention is not more than 5 days after the operation. The term "catabolic change after surgery" is generally used [Shaw,, Η.Ρ., ^ Λη, Sur, (1989); Ut ^ l ^ (1987); Frayn, KK · N. (1986); Brandi (L., et al., (1993)], and as defined in this paper as the metabolic state caused by surgical trauma, can be characterized by one or more of the following phenomena: negative nitrogen balance, body nitrogen loss. This paper is applicable to China. Standard (CNS) A4 specification (210XM? Mm)

• —1— ....... 1 ml. I i ----- pack --- (please read the notes on the back of the book first) HI ID · --- line 492869 A7 B7 V. Invention Explanation (25 Printed by Shao Central Standard Bureau Staff Consumer Cooperatives [Wernerman, J ·, et al., J. Parent. Enter. Nutr 1 η · * u * ^ / 8-82 (1986); Tashiro, T ·, Etc., J. Parent. Enter. Nutr, 9:, ^ -2-5 (19S5)], the use of fat around the perimeter is better than glucose plus respiratory quotient [Fiayn, κ.Ν ·, etc., Arch Emerg. Med, 4: 91-9 (1987) Stjernstrom, H., et al., Clin. Physiol. 1-. 59-72 (198 i)], and consumes body protein and energy despite hyperglycemia Storage for endogenous glucose production [Gump, FE, et al. (1974); Black, R. B. et al. (1982), Frayn, K · N ·, et al. (1987); Frayn, K · N · BΓ · Med Bull. 41 (3): 232-9 (1985)] 〇 The term "insulin resistance" is also well known to general physicians, and is defined herein as a normal concentration of insulin that causes a physiological condition that is less than the normal response. Resistance may be due to insulin and cell surface receptors Combined reduction, or changes in intracellular metabolism. The first type is characterized by decreased insulin sensitivity and can typically be overcome by increasing insulin concentration. The second type is characterized by decreased insulin reactivity that cannot be overcome even with large amounts of insulin. Post-traumatic Tengdaosu resistance can be overcome by adding only insulin proportional to insulin resistance '. Therefore, it is obviously caused by a decrease in insulin sensitivity [Brandi. LS ·, et al., Clin · Science 79: 443-450 ( 1990); Henderson, Α · Α ·, et al., Clin · Sci · 80: 25-32 (1990)]. Insulin sensitivity decreases after selective abdominal surgery for at least 5 days, but not more than 3 weeks, and the surgery The latter part is the most severe and may take three weeks to normalize. [ThorreU, A ·, et al. (1 993)]. The cause of insulin resistance seen in trauma is still unknown. Patients who want to normalize their blood glucose "GLp The effective dose of —i, glp-1 analogue or GLP-1 non-biological organism depends on a number of factors, including but not limited to, please read first.

492869 A7 B7 V. Description of the invention (26. The Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs printed the gender, weight and age of the person, unable to regulate the severity of blood glucose, the cause of unregulated blood glucose, whether to apply glucose or other sugar sources at the same time Route of administration and bioavailability, in vivo persistence, formulations and potency. For continuous administration, suitable dosage rates are 0.25 to 6 picomoles / kg weight / knife, 0.5 to about 1.2 picomoles / Kg / min is preferred. When intermittently administered, each application dose should consider the application interval, the bioavailability of GLP-1, GLp-1 analogs or GLP-1 derivatives and the amount required to achieve normal blood glucose. General doctor The technology of administering dosage and rate of Gpu, GLP-1 analogue or GLP-1 derivative which can achieve the required clinical result is developed by the handy kit. The present invention is easier to understand with reference to specific examples, but it is only for illustration and not limitation of the present invention. Examples 1 This study involved 13 patients who wanted to undergo selective orthoplasty. No one had a history or sign of metabolic disease, liver disease, or diabetes. Blood glucose during fasting in all people , C Rp, and liver tests (bilirubin, alkaline phosphatase, AST, and ALT) were normal. Seven patients were studied at 08:00: 00 in the morning after overnight fasting (insulin group, age, 56 ± 5 years old) , BMI 25 ± 1 kg / m 2). After the initial reference period, the blood glucose and hormones were sampled and the indirect energy was measured for 30 minutes, and insulin was injected at a fixed rate of 0.8 milliunits / kg / min (Actrapid ®, Novo, Copenhagen), while intravenously injecting a variable amount of glucose (200 mg / ml) to maintain blood glucose at a fixed level (4.5 mg mole concentration). One hour after this stable condition, all patients were standardized hand treatment (Hipartroplasty). Surgery is performed at 290 to 23 minutes after Takishima prime infusion. Please read it first. T & Note · This page binding clock 29- This paper size applies Chinese National Standard (CNS) A4g (210X297 mm) ) Printed and described by the Consumers' Cooperative of the China Prospective Bureau (27 Kai Shikou. Cone during surgery ^ Follow-up d Ϊ Return blood insulin, normal blood sugar control, and during surgery. Information It will be presented according to the following names: 30 minutes before insulin perfusion, surgery for 60 minutes of stable hyperglycemia, normal blood glucose control after the start of surgery, i to 40 minutes, the last 30 minutes of surgery, and 143 to 30 minutes of surgery. 60 minutes high _ Insulin, normal blood glucose control type 2 patients (for daily g 4 and & & 〆 ,, / 丁 ~ ,,, and n-6, age, 59 ± 3 years old, BMI 26: tl Control of the pre-operation period in the baseline period. Pre-operation control period. Pre-operation period and post-operation control items. 8 V 7 丄 j wind, JD iVi 丄 2 D 3 1 million meters. Regarding the age and the coordination with the Tengdaosu group. In hand: Accept the same in 7 days. Pre-operative methods (baseline and pre-operative controls). Dui Teng, did not receive baseline or pre-operative control during surgery. Although, in the immediate operation group, every patient in the group received insulin perfusion (.8 milliunits / kg / min) and started hyperglycemic, normal blood glucose control (4 $ millimolar control (after surgery). Indirect calories Assay (DeltatracVDansj00, ".η") KNJ Appl. Physiol. 55 (2): 628-34 (i983); Takala, J · 'et al., Crit. Care Med · 17⑽: 1041_47 (i989)] is The baseline period, two times during the operation (before and after the operation) were performed for 30 minutes each, and the last 30 minutes of the control before and after the operation. Urine sampling was also performed regularly to analyze the urea excretion. The urea concentration was corrected Changes [Tappy, L, etc .., Diabetes 37 · 1212-16 (1988) j, then calculate non-protein energy expenditure (EE), respiratory quotient (RQ), and substrate oxidation rate. Shu Zhaoding 30- this Paper size applies to Chinese National Standard (CNS) A4 (2I0X297 mm) 492869 A7 B7 V. Invention Description (28 Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs

Blood samples were collected repeatedly from heated palm veins during the baseline, pre-operative, pre-operative, post-operative, and post-operative periods. Immediately after collection, blood glucose was measured using the glucose oxidase method (Yellow Springs Instruments, Yellow Springs, Ohio) [Hugget, A.S., et al., Lancet 2: 368 Lai 70 (1957)]. Determination of serum insulin concentration using radioimmunoassay (RIA) [Grill, V., et al., Metabolism 39: 25 1 -5 8 (1 990)]; C-peptide (Novo Research, Bagsverd, Denmark); corticosteroids [ Harris, V., et al., In Jaffe, BM &amp; Behrman, HR, ed., Methods of hormone radioimmunoassay, Academic Press, New York and London (1979) pp. 043-50]; and glucagon ( Euro-Diagnostica AB5 Malmb, Sweden) [Faloona, GR, et al., Glucagon radioimmunoassay technique. Volume 1: Academic Press, New York (1974)] 〇 All are individual groups or mean ± SEM (standard deviation of the mean). The statistically significant acceptor was ρ <0.05, which used Wilcoon, s famous comparison test and Mann-Wlntney U-test on paired and unpaired data, respectively. Because the amount of serum vasopressin in the control period after surgery was lower than that in the control group (P = 0.06), we distinguished the gir stage from the 60-minute stable gir and the mean blood island level. GIR is corrected to a general insulin amount. The serum levels of the two groups were similar in both groups, regardless of whether they were controlled during the baseline period or before the operation. In the control group, the Tengdaosu amount remained unchanged from the baseline amount during surgery. The control group ’s post-operative control period was not significantly different from the control period, and the control period was not significantly different from that of the control group during the post-operative control period. It was also appropriate to control the Z 'paper size (CNS). (Please read the precautions on the back. ·-: Write this page) -Pack. Line -31-492869

7 B V. Invention Note (29) There is no difference. During pre- and post-operative control periods, the amount of c-peptide at baseline was similar between the two groups (Table I). In comparison to the control group, the osmoticin group showed a lower amount of peptide during surgery. After surgery, the levels of serum shizatin decreased significantly (p &lt; 0.05) (Table I). However, the relative change after surgery (%, before surgery) was higher in the insulin group (p &lt; 0.01 vs. control group). The amount of serum corticosteroids (Table I) decreased after surgery in the insulin group, while the amount tended to increase in the control group (ρ = 0.1). Compared with the control group, the insulin group had lower corticosteroids after surgery (ρ &lt; 0.05). (Please read the notes on ¾. Please write this page.) Binding and printing. Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs. I. Patients undergoing orthopedic surgery are fasted overnight (control group, η = 6), or _ After 4 hours, the hormone amount of physiological hyperinsulin (Tengdaosu, η = 7) was controlled before surgery. The pre-operative control was controlled by the C-peptide control group after the postoperative operation. 68 ± .08 .41 ± .09 .70. 11 .70 ± .13 .31 ± .09 Insulin 68 ± .09 .45 ± .05 • 42 ± .0 + amp; • 58 ± .12 J2 ± .11 Glycein control group 48 ± 2 42 ± 1 43 ± 3 41 soil 3 37 soil 2 * insulin 58 ± 7 52 ± 31 40 ± 3 35 ± 4 33 ± 4 * corticosteroid control group 229 ± 39 238 ± 21 154 soil 63 116 soil 43 366 ± 83 * 41 266 ± 35 234 ± 46 212 ± 44 172 Soil 83f * * ρ &lt; 0 · 0 5 using Wi i 1 c ο X ο η signature comparison test compared with before operation ten ρ &lt; 0.05 using Mann-Whitney U-test and Comparison line of the control group -32- This paper size is applicable to China National Standard (CNS) A4 specification (210X 297mm) 492869

Description of the invention (3Q: Glycogen levels in both groups decreased after surgery, and the larger group was found to be larger (p &lt; 0.01 vs. control group). Skin decline (P &lt; 0. G5 was controlled by insulin before surgery) In the group, "after surgery", Tengdao after surgery: / :, and the tendency to increase in the control group was significantly lower (p &lt; 0 05). There is no significant difference to (㈣). Compared with the control period before surgery ==: &lt; The control group during the second-hand postoperative control period has a decrease in mean GIR. The control group maintains GIR during surgery. 9Λ. The control period after surgery even tends to improve coffee (+ 16 ±, / °: P: 〇 · 2). The most significant and unexpected, the control group controls the hand after surgery: two: the control group has a significantly higher ratio (P &lt; ° · 05) (refer to Figure 1). All changes in the control period and the post-operative control period = P &lt; ... 5), regardless of whether the average serum Tengdaosu amount is stable in the stable period. The rate of glucose and fat oxidation was similar between groups. During surgery, the rate of glucose oxidation in the heart sentence of the prime group was significantly higher, and the lipid training was significantly lower (P &lt; 0 · 05 for the control group). During the post-operative control period, the plate = then the control period: no change in the pancreatic "oxidation rate of the pancreas" group. T = or there was no difference in the resting energy consumption (EE) between the two groups after the operation Compared with the control period before and after surgery, the results of the two groups were still the same. The glucose amount was similar between the insulin group and the control group when fasting. In the meantime, it is because maintaining the normal blood borrow concentration caused the individual changes in the control group. The coefficient is 4.6%, while the insulin group is 6.2%, and the forehand is -33- this paper CNS) A4 size γ ^ 297 public 492869 printed by the staff consumer cooperative of the Central Standards Bureau of the Ministry of Economy -34- A7 B7 5 Explanation of the invention (31) The conclusions obtained from these findings show that patients undergoing selective surgery in the fasting state will develop increased insulin resistance and fat oxidation after surgery. Moreover, these findings also show for the first time that if a patient is in a state of hyperinsulinism during surgery and is maintained during surgery, the alienation changes after surgery will be completely eliminated, and the hormonal response to compression will be completely weakened. Example 2 GLP-1 (7-3 6) Si amines were taken at 2 picomoles / kg / hour at a dose rate of 10 hours subcutaneously at night for five non-insulin-dependent diabetic patients (NIDDM). As a control, insulin was continuously infused into the same five patients, but on a different day from the infusion of GLP-1 (7.36) amidine. Adjust the rate of insulin perfusion every two hours so that hypoglycemia is optimally controlled and avoided. As shown in the data in Table II and Figure 2, subcutaneous perfusion of GLp_1 (7_36) 醯 Yuewen went to normalize blood glucose without defaming hypoglycemia in any patient. GLP-1 (7-3 6) amidamine was used for metabolic control better than insulin, and the average blood glucose of GLP-1 (7-36) amidamine was 23:00 compared with the control group. At 0:00 and 1:00, the statistics are significantly lower. ° This paper size is suitable for 囫 囫 囫 擭 quasi r mtc, Λ4 said Christine (210, 乂 297 / \ chrome

492869 A7 B7 Printed by the Consumer Cooperative of the Central Bureau of Standards, Ministry of Economic Affairs, and a description of the invention (32 Table π · The average blood glucose of 5 NIDDM patients at 10 hours of continuous perfusion with GLP-1 (7-3 6) amide for 10 hours. The same patient was performed on different days. ^ _ In the control group, insulin was administered by continuous infusion. _. GUM infusion insulin infusion _______ hours mean standard deviation of blood glucose ~~ Μψί ^ — one (millimolar concentration 1 (millimolar concentration (Concentration of ear) (concentration of mole) 21:00 7.5 0.45 6.9 0.68 22:00 5.4 0.76 6.6 0.55 23:00 4.1 0.16 5.9 0.98 0:00 4.4 0.23 5.6 0.90 1:00 4.4 0.29 5.1 0.58 2:00 4.8 0.34 5.2 0.58 3:00 5.2 0.41 5.4 0.30 4:00 5.4 0.41 5.7 0.25 5:00 5.8 0.41 6.0 0.30 6:00 6.0 0.45 6.1 0.38 7:00 6.2 0.45 6.1 0.33 Example 3 The day of perfusion was performed for five persons at three meals NIDDM patients were perfused for 3 hours. The perfusion time is shown in Figure 3 as 7: 3 0-1 0: 3 0 (breakfast), 10: 3 0 -1: 3〇 (lunch) and 4: 3〇_7: 3 〇 (dinner). Control group experiments on the same patient on different days' as shown in Figure 3, subcutaneous injection of insulin before three meals. Note GLP-1, glucose due to insulin injections after findings from the China National Standards dinner executive sentiment (492,869 patent application No. 86112441 Chinese manual correction pages (89 February) V. description of the invention (33 A7 Βί7

GLP-1 perfusion hour Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs' to reduce normal blood glucose levels. After termination of each G l p -1 (7-3 6) amidine injection, the blood glucose level increased significantly. No adverse effects of G l p-1 (7-3 6) amide were observed. These data show that GLP-1 (7-36) ammonium perfusion can more effectively control the amount of glucose after meals than insulin perfusion, and it is effective as long as the control of GLP-1 (7-3 6) perfusion continues. Table 111. The average blood glucose level of 5 NIDDM patients infused with g L P -1 (7-36) amines 3 hours before the start of each meal. In the control group performed on the same patient on different days, 'Insulin is injected subcutaneously before each meal' / _ Meal start time is 7: 3 0, 1 0 · 3 0 and 16: 3 0_ Tengdaosu subcutaneous injection Mean blood glucose standard deviation Mean blood glucose standard (millimolar concentration) (millimolar concentration) (millimolar concentration) (millimolar 7:00 5.4 0.35 6.1 0.41 8:00 4.9 038 7.0 0.51 9:00 5.7 0.59 9.1 0.74 10:00 5.8 L06 9.9 0.78 11:00 8.1 0.94 8.2 0.76 12:00 9.4 0.59 6.5 0.74 13:00 7.2 1.18 9.1 0.90 14:00 5.3 1.21 8.1 0.91 15:00 7.2 0.71 7.0 0.87 16:00 10.4 0.26 12 0.57 17 : 00 9.2 L06 6.5 0.59 -36- This paper size applies to China National Standard (CNS) A4 specification (210X297 mm) 492869 A7 B7 V. Description of the invention (34) 18:00 5.7 1.59 7.3 0.65 19:00 6.6 0.94 6.1 0.59 20:00 8.3 0.71 6.0 0.41 21:00 9.3 0.71 6.4 0.44 (Precautions for reading and reading on the back of the image ^ Write this page) Binding and threading Printed by the Staff Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs-37- Paper size applies to China National Standard (CNS) A4 (210X297 mm)

Claims (1)

Patent No. 086112441 Chinese Patent Application Amendment (April 91) 1. A pharmaceutical composition for attenuating catabolic changes and insulin resistance after surgery ', comprising a compound selected from GLP-1, GLP-1 analogs, GLP-1 derivatives, and pharmaceutically acceptable salts thereof. 2. The pharmaceutical composition according to item 丨 of the application, wherein the compound is administered intravenously. 3. The pharmaceutical composition according to item 1 of the application, wherein the compound is administered subcutaneously. 4. The pharmaceutical composition according to item 2 or 3 of the scope of patent application, wherein the administration method is continuous. 5. The pharmaceutical composition according to item 4 of the application, wherein the compound is applied at a rate of between 0.25 and 6 picomoles / kg / min. 6. The pharmaceutical composition according to item 5 of the application, wherein the compound is applied at a rate of 0.5 to 2.4 picomoles / kg / min. 7. The pharmaceutical composition according to item 5 of the application, wherein the rate is between about 0.5 to about 1.2 picomoles / kg / min. 8. The pharmaceutical composition according to item 2 of the application, wherein the intravenous administration is intermittent. 9. The pharmaceutical composition according to item 2 of the scope of patent application, wherein the compound is administered intravenously, but also by other parenteral routes. 10. The pharmaceutical composition according to item 9 of the application, wherein the other parenteral route is a subcutaneous route. 11. The pharmaceutical composition according to item 1 of the scope of patent application, wherein the compound to be administered is GLP (7-36) amidamine or a pharmaceutically acceptable salt thereof. This paper size applies to China National Standard (CNS) A4 (210 X 297 mm)