TW201318636A - GLP-1 agonist for use in the treatment of stenosis or/and obstruction in the biliary tract - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for use in the treatment of a disease or condition, wherein said disease or condition is associated (a) stenosis or/and obstruction located in the biliary tract, or/and (b) biliary dyskinesia, said composition comprising at least one GLP-1 agonist and optionally a pharmaceutically acceptable carrier, diluent or/and auxiliary substance.

Description

GLP-1 agonist for the treatment of biliary strictures and/or obstructions

The present invention relates to a pharmaceutical composition for treating a disease or condition, wherein the disease or condition is associated with (a) stenosis or/and obstruction in the biliary tract, or/and (b) biliary dyskinesia, The composition comprises at least one GLP-1 agonist and optionally a pharmaceutically acceptable carrier, diluent or/and auxiliary.

The biliary tract (also known as the biliary or bile duct system) introduces bile from the liver into the duodenum. The biliary tract includes ductus hepaticus (hepatic duct, common hepatic duct), ductus cysticus (cystic duct), gallbladder, ductus choledochus (common bile duct), hepatic pancreatic duct, papilla of Vater or / and Oddi's sphincter. The total hepatic duct leads to the common bile duct. The common bile duct is formed by the confluence of the total hepatic duct and the cystic duct. The cystic duct connects the gallbladder to the hepatic duct/general bile duct. The hepatopancreatic duct is formed by the confluence of the common bile duct and the large pancreatic duct, producing a pedicle of the genus of the duodenum (also known as the nipple or duodenal papilla). The lack of nipples includes the Odyssey sphincter. The Odyssey sphincter is composed of smooth muscle fibers surrounding a variable length hepatic pancreatic duct.

The biliary wall contains smooth muscle and the inner surface is covered with a columnar epithelium.

Smooth muscle contraction in the biliary tract (especially the gallbladder) can be caused by cholecystokinin (CCK). CCK is produced in the small intestinal mucosa. CCK reaches the gallbladder via circulation. CCK release may be caused by a fat meal. through After the gallbladder contracts, the bile is released into the duodenum that promotes fat emulsification. Another function of CCK is to induce the release of digestive enzymes in the stomach and pancreas. Stenosis or/and obstruction in the biliary tract may result in a decrease in bile flow, resulting in a condition associated with pain. Therefore, the inflammatory process continues the pathophysiological condition. For example, stenosis or/and occlusion in the biliary tract due to ingrowth of surrounding tissue (because of cancer or/and inflammatory processes) can lead to increased bile accumulation and increased tube volume, combined with wall dilation. An increase in volume combined with diastolic wall may result in severe acute or chronic pain. Acute pain is sometimes referred to as the "onset" of the gallbladder because of its sudden onset. Treatment of such pain includes treatment with an opioid analgesic or surgical removal of stenosis or/and occlusion. In addition, the surgery can be a "radical" surgery, that is, completely removing the rickets tissue (such as the gallbladder).

Blockage may be caused by stones such as cholesterol stones, bilirubin and calcium salt stones, or "mixed" stones containing cholesterol and bilirubin. These stones can also be called gallstones. The appearance of stones in the bile duct is called choledocholithiasis. The occurrence of stones in the gallbladder is called cholecystolithiasis.

Biliary stenosis may also be caused by cancer or other processes that cause the growth of the target tissue (such as exudative inflammatory processes).

Glucagon-like peptide-1 (GLP-1) is an endocrine hormone that increases insulin response after oral intake of glucose or fat. GLP-1 usually regulates glycemic concentration, slows gastric emptying, stimulates (original) insulin biosynthesis, increases insulin sensitivity, and stimulates insulin-dependent biosynthesis of glycogen (Holst (1999), Curr. Med.Chem 6:1005, Nauck et al. (1997) Exp Clin Endocrinol Diabetes 105: 187, Lopez-Delgado et al. (1998) Endocrinology 139:2811).

Human GLP-1 has 37 amino acid residues (Heinrich et al., Endocrinol. 115: 2176. (1984), Uttenthal et al., J Clin Endocrinol Metabol (1985) 61:472). Active fragments of GLP-1 include GLP-1 (7-36) guanamine and GLP-1 (7-37).

Exendin is a group of peptides that lower blood sugar levels. Exenatide has some similarity in sequence to GLP-1 (7-36) (53%, Goke et al. J. Biol Chem 268, 19650-55). Exenatide-3 and Exendin-4 stimulate the increase in cellular cAMP production by interacting with the Exenatide receptor in acinar cells of the guinea pig pancreas (Raufman, 1996, Reg. Peptides 61:1- 18). Exenatide-3 causes an increase in amylase release in acinar cells of the pancreas relative to exenatide-4.

Exenatide-3, Exendin-4 and Exenatide agonists have been proposed for the treatment of diabetes and prevention of hyperglycemia; they reduce gastric motility and gastric emptying (US 5,424,286 and WO98/05351) .

Exenatide analogs can be identified based on amino acid substitution and/or C-terminal truncation of the native Exendin-4 sequence. Such exenatide analogs are described in WO 99/07404, WO 99/25727, WO 99/25728. Exenatide-4 analogs include lixisenatide (also known as AVE0010, desPro ³⁶ - Exenatide-4-Lys ₆ -NH ₂ or H-desPro ³⁶ - Exenatide-4- Lys ₆ -NH ₂ ).

WO 2007/028394 discloses the use of GLP-1 molecules for the treatment of biliary dyskinesia and/or biliary pain/discomfort. According to WO 2007/028394, biliary tract The dyskinesia may be an increase in movement in the biliary region or a decrease in movement in the biliary region. The GLP-1 molecule is considered to be a peristaltic promoter. According to WO 2007/028394, GLP-1 molecules can be administered in combination with one or more other stimulating factors (etc.) capable of causing bile flow and/or treating biliary motility disorders. Alternatively, the GLP-1 molecule can be administered in combination with one or more inhibitory factors that reduce bile flow. No experimental validation of GLP-1 molecules affecting the physiology of gallbladder is presented in WO 2007/028394. Unknown GLP-1 has any effect on the biliary tract.

Gallbladder movement can be expressed as a gallbladder ejection fraction and can be assessed by cholangiography. Biliary scintigraphy is a nuclear imaging step to assess gallbladder function. For example, an imine diacetic acid derivative labeled with ^{99 m} quinone is injected iv and then allowed to circulate to the liver where it is secreted to the bile system and stored by the gallbladder. Biliary scintigraphy is used to diagnose biliary dyskinesia, which involves gallbladder dysfunction. Cases with a gallbladder excretion rate (GBEF) of less than 40% are considered to represent cholecystectomy (Behar J. et al., Gastroenterology 2006 130: 1498-1509).

Relaxation and contraction of the gallbladder is a type association that is mediated by smooth muscle cells and exhibits dynamic measurements via ^99m biliary scintigraphy.

Summary of invention

An embodiment of the invention is a randomized, double-blind, placebo-controlled, double-sequence, dual-treatment crossover study evaluating the efficacy of a single subcutaneous injection of 20 μg lixisenatide for gallbladder movement in healthy male and female subjects. Gallbladder movement has been analyzed by cholangiography. Cholecystokinin (CCK-8) has been administered 60 minutes after administration of a single dose of placebo or 20 μg of lixisenatide, followed by a single dose of ^99m Tc mebrofenin (a ^99m Tc marker) Imine diacetic acid derivative). The procedure can be summarized as follows. By placing a radiation sensitive camera on the subject's abdomen, an "image" of the liver, bile duct and gallbladder is obtained, which corresponds to where the radioactive bile moves. The liver frame map was obtained at 1 frame/minute for 60 minutes after injection of lixisenatide or placebo with ^99m Tc injection. After a gallbladder view at 60 minutes, 0.02 μg/kg CCK-8 was administered via a continuous infusion pump for 60 minutes. The image was captured for at least another 60 minutes after the CCK-8 infusion.

In the placebo group, CCK-8 caused a decrease in the count recorded by the gallbladder, indicating that bile was released from the gallbladder via the bile duct into the duodenum. Surprisingly, this effect of CCK-8 was greatly reduced by subcutaneous administration of a single dose of 20 μg lixisenatide in healthy subjects (see the exemplary filling curve and emptying curve in Figure 2). ). The ^99m Tc in the gallbladder is maintained at a high level, indicating that only a small portion of the bile is released. This utility has been quantified as follows. In the placebo group, CCK-8 caused a gallbladder excretion rate (GBEF, or excretion rate EF, which is the bile volume fraction excreted from the gallbladder) to increase by up to about 85%, as expected from the physiological effects of CCK. In the lixisenatide group, the excretion rate was maintained below 40% after CCK-8 administration (see the exemplary GBEF curve in Figure 3, the average is provided in Figures 7 and 9). Subject to lixisenatide, 13/24 (54%) subjects had a GBEF of less than 40% at 60 minutes compared to 1/24 (4%) of placebo. Subcutaneous administration of a single dose of 20 μg lixisenatide in healthy subjects significantly reduced gallbladder emptying (expressed as gallbladder excretion rate (GBEF)) caused by CCK-8 response compared to placebo at 60 45.8% in minutes.

As described herein, the reason for the utility of Lisila to GBEF is unknown. Without wishing to be bound by theory, the reduction of gallbladder emptying caused by CCK due to lixisenatide may be interpreted as lixisenatide to cause smooth muscle relaxation of the gallbladder. Therefore, Lisila comes to provide antispasmodic effects in the gallbladder. We can infer that lixisenatide can exhibit smooth muscle relaxation in the bile system. Based on the utility of lixisenatide for GBEF, it is expected that smooth muscle cells in the biliary system will become slack due to administration of lixisenatide or GLP-1 agonist. Therefore, GLP-1 agonists, including lixisenatide, can provide antispasmodic or antispasmodic effects in the bile system.

Stenosis or/and obstruction in the biliary tract can cause paralysis and usually pain. The antispasmodic utility of GLP-1 agonists suggests the therapeutic use of GLP-1 agonists in the treatment of diseases associated with stenosis or obstruction processes. GLP-1 agonists, such as lixisenatide, may exhibit therapeutic utility for stenosis or occlusion of the biliary tract. Smooth muscle relaxation increases the lumen and thus increases bile flow if stenosis or obstruction occurs. Because of its antispasmodic activity, GLP-1 agonists have become suitable for the treatment of pain associated with stenosis or/and occlusion in the bile system.

Accordingly, a first aspect of the invention is a pharmaceutical composition for treating a disease or condition, wherein the disease or condition is associated with (a) stenosis or/and obstruction in the biliary tract, or/and (b) biliary tract In connection with dyskinesia, the composition comprises at least one GLP-1 agonist and optionally a pharmaceutically acceptable carrier, diluent or/and auxiliary substance.

In the present invention, "biliary tract" includes, but is not limited to, hepatic duct (ductus) Hepaticus), gallbladder, ductus cysticus, ductus choledochus, hepatopancreatic duct, papilla of Vater or/and Oddi's sphincter.

As used herein, "hepatic duct" or "hepatic duct" includes the right hepatic duct and the left hepatic duct, as well as the total hepatic duct. The total hepatic duct is formed by the confluence of the right hepatic duct (the bile from the right functional leaf of the liver) and the left hepatic duct (the bile from the left functional lobe of the liver). In particular, "hepatic duct" or "hepatic duct" means those portions of the hepatic duct that are outside the liver tissue.

Stenosis or/and obstruction may be in the hepatic duct, in the gallbladder, in the cystic duct, in the common bile duct, in the hepatic pancreatic duct, in the nipple or in the eddy sphincter.

Blockage may be caused by stones, stone disease, and/or biliary mud in the biliary tract. Specifically, stone disease includes gallstones and cholelithiasis. Stones may be cholesterol stones, pigment stones or stones containing cholesterol and pigments ("hybrid" stones).

In the present invention, the cholesterol stone or the cholesterol stone may comprise at least 80% by weight of cholesterol, or at least 70% by weight of cholesterol.

In the present invention, the pigment stone or pigment stone may contain bilirubin or/and a calcium salt, wherein the calcium salt is found in the gall bladder (for example, calcium hydrogencarbonate). Pigment stones may comprise up to 20% by weight, or up to 30% by weight of cholesterol.

In the present invention, the mixed stone or mixed stone may contain 20 to 80% by weight of cholesterol or 30 to 70% by weight of cholesterol. Mixed stones may contain calcium carbonate, palmitate phosphate, bilirubin, Or / and other bilirubin.

"Bile mud" as used herein, refers to a mixture of microscopic particulate materials that appear in the gallbladder when the material particles are precipitated from the gallbladder. The particulate material may comprise the same ingredients as the stones described herein. The most common particle components of bile mud are cholesterol crystals and calcium salts.

Some diseases are associated with biliary pain and/or discomfort and/or dyskinesia (or "biliary dyskinesia"). A "dyskinesia" in the biliary tract is defined herein as an abnormality in movement of any part of the biliary tract, such as, for example, gallbladder, bile duct wall, cystic duct, Odyssey sphincter, and/or abnormal motion of the gallbladder sphincter. In particular, abnormal biliary motility causes pain and/or discomfort to the patient. This includes, but is not limited to, abnormal gallbladder function, biliary dysfunction, and dysfunction of the Odyssey sphincter. Abnormal biliary dysfunction may be an increase in movement of a certain area of the biliary tract, or a decrease in movement of a certain area of the biliary tract, or an dysregulation of movement such as, for example, biliary fistula.

In the present invention, abnormal gallbladder function may be an abnormal movement of the gallbladder, including abnormal gallbladder emptying, which causes gallbladder pain or discomfort.

In the present invention, Oddi's sphincter dysfunction is a term used to define the abnormality of the Odyssey sphincter.

Diseases associated with stenosis or/and obstruction of the biliary tract may be pain or discomfort and/or dyskinesia ("or biliary dyskinesia") in the biliary tract. A disease associated with stenosis or/and occlusion of the biliary tract is specifically the disease disclosed in WO 2007/028394, the disclosure of which is incorporated herein by reference. The disease may be biliary dyskinesia and/or pain or discomfort derived from the biliary tract, and has at least one pathological condition selected from the group consisting of inflammatory bowel disease, chronic acalculous cholecystitis (acute acalculous) Cholecystitis), acalculous gallbladder disease, cystic duct syndrome, post-cholecystectomy syndrome, treatment with gallbladder movement inhibitory drugs, intravenous nutrition, celiac disease, Down's syndrome, beta thalassemia, stomach surgery, very low calorie diet , extracorporeal shock wave lithotripsy, growth inhibitory sphincter, eddy sphincter stenosis, eddy sphincter dyskinesia, glandular hyperplasia, muscle hyperplasia, muscle hypertrophy, muscle fibrosis, eddy sphincter muscle dysmotility, audi Sphincter muscle hypertonia, Odyssey sphincter muscle spasm, hyperthyroidism, use of compounds known to interfere with bile passage (such as somatostatin, octreotide, sandostatin LAR, anticholinergic Treatment of drugs, nitric oxide, L-arginine, nitric oxide donors, calcium channel antagonists, biliary obstruction (partial or complete), biliary obstruction caused by biliary stones (partial or complete) ), cholangiocarcinoma, primary sclerosing cholangitis (PSC), pediatric ampullary tumor, other types of biliary tumors (including papilloma, adenoma, and cyst Tumor), Mirizzi's syndrome, AIDS cholangiopathies, infections (including Cryptosporidium, cytomegalovirus, microsporidia, and spore coccidia), parasitic infections (such as worms, mites, and liver lice, such as Chinese worms) And liver fluke), obstructive lesions in the large tube outside the liver, primary biliary cirrhosis, primary sclerosing cholangitis, obstructive biliary tract disease in infants (eg extrahepatic biliary atresia), Because of drugs such as phenoxaprofen, chlorpromazine, fluridol, imipramine, damage to the intrahepatic bile duct, bile and bile stasis.

In the present invention, a typical nitric oxide donor may be glyceryl trinitrate or nitroprusside. The calcium channel antagonist may be selected from the group consisting of nifedipine, verapamil, loperamide, and lutein. (progesterone), trimebutine maleate, loperamide and ondansetron.

Diseases associated with stenosis or/and obstruction of the biliary tract may be biliary dyskinesia and/or pain or discomfort resulting from the biliary tract associated with gallstones. Pain or discomfort and/or movement disorders may occur anywhere in the individual's biliary tract. The disease associated with stenosis or/and obstruction may be pain and/or discomfort and/or dyskinesia in one or more of the following: gallbladder, Oddi's sphincter, bile duct wall, and/or cystic duct.

A disease associated with stenosis or/and obstruction of the biliary tract may be a pathological condition that causes gallbladder dyskinesia and/or pain and/or discomfort, wherein the pathological condition is preferably selected from the group consisting of biliary dyskinesia, gallstone, and inflammatory bowel disease. Chronic acalculous cholecystitis, acalculous gallbladder disease, cystic duct syndrome and post-cholecystectomy syndrome.

A disease associated with stenosis or/and obstruction of the biliary tract may be a pathological condition associated with causing gallbladder dyskinesia and/or pain and/or discomfort, wherein the pathological condition is preferably selected from the group consisting of treatment with a gallbladder movement inhibitory drug, intravenous nutrition. , celiac disease, Down's syndrome, beta thalassemia, gastric surgery, very low calorie diet, extracorporeal shock wave lithotripsy and somatostatin to produce tumors.

A disease associated with stenosis or/and obstruction of the biliary tract may be a pathological condition that causes Odyssey sphincter dyskinesia and/or pain and/or discomfort. Thus, for example, the individual may be affected by "Oddi's sphincter stenosis" (eg, due to glandular hyperplasia, muscle hyperplasia, muscle hypertrophy, or muscle fibrosis), and/or "Oddi's sphincter dyskinesia" (eg, by muscle movement) Suffering from dysregulation, excessive muscle tension or paralysis.

A disease associated with stenosis or/and obstruction of the biliary tract may be an Odd's sphincter dyskinesia in which the individual suffering from it has an abnormal contraction of the Odyssey sphincter 7 or more times per minute. Additionally, Oddi's sphincter dyskinesia and/or pain and/or discomfort may involve abnormalities such as the sphincter, pancreatic sphincters, or both. Oddi's sphincter dyskinesia may be associated with hyperthyroidism.

A disease associated with stenosis or/and occlusion of the biliary tract may be a pathological condition resulting from the treatment with a therapeutic factor (such as a drug) known to inhibit biliary tract movement and/or reduce the underlying tension of the bile system. The inhibitory factor can be, but is not limited to, any of the following factors: somatostatin, octreotide, serotonin LAR, anticholinergic drugs, nitric oxide, L-arginine, nitric oxide donor, and calcium. Channel antagonist.

Other biliary tract disorders that may be treated in accordance with the present invention that cause biliary dyskinesia and/or pain or/and discomfort include, but are not limited to, biliary obstruction (partial or complete), bile caused by biliary stones Obstruction (partial or complete), cholangiocarcinoma, primary sclerosing cholangitis (PSC), pediatric ampullary tumor, other types of biliary tumors (including papilloma, adenoma and cystadenoma), Mirizzi's syndrome , AIDS cholangiopathies, infections (including Cryptosporidium, cytomegalovirus, microsporidia, and spore coccidia), parasitic infections (such as worms, mites, and liver lice, such as Chinese worms and liver flukes), Obstructive lesions of biliary trees in large tubes outside the liver, primary biliary cirrhosis, primary sclerosing cholangitis, obstructive biliary tract disease in infants (eg extrahepatic biliary atresia), intrahepatic bile ducts Injury (because, for example, drugs such as phenoxaprofen, chlorpromazine, fluridol, imipramine, etc.), bile and bile stasis.

Pregnancy may be associated with stenosis or/or obstruction of the biliary tract.

In particular, diseases associated with stenosis or/and obstruction in the biliary tract may be calculi associated with stone disease, gallstones, cholelithiasis, or/and formation of stones in the biliary tract.

A disease or condition associated with stenosis or/and obstruction in the biliary tract can be cancer. Specifically, cancer causes stenosis in the biliary tract.

The cancer can be in the hepatic duct, in the gallbladder, in the cystic duct, in the common bile duct, in the hepatic pancreatic duct, in the nipple or in the eddy sphincter. In particular, cancer includes tumors. Cancer can include metastasis, such as a metastasis derived from a metastatic cancer in another organ.

Stenosis or/and obstruction may cause pain.

Treatment can be a palliative treatment. Mitigation is necessary in patients with cancer, especially inoperable cancer, terminal cancer, cancer that does not respond to cancer treatment (eg due to radiation or/and chemotherapy), there is a shift from another organ Metastasis derived from sexual cancer, wherein metastasis results in stenosis or/and occlusion in the biliary tract, and wherein the cancer is specifically painful, in particular because of the formation of stenosis or/and occlusion. More specifically, cancer forms a stenosis.

Treatment, especially palliative treatment, can last for at least one month, at least two months, at least three months, at least four months, or at least six months.

Another aspect of the invention is a pharmaceutical composition for palliative treatment comprising at least one GLP-1 agonist, and optionally a pharmaceutically acceptable carrier, diluent or/and auxiliary substance. The palliative treatment is the palliative treatment described herein. In particular, palliative treatment includes treatment of pain caused by stenosis or/and obstruction in the biliary tract. pain. More specifically, palliative treatment includes treatment of stenosis of the biliary tract and/or pain of cancer caused by occlusion. The composition is a composition as described herein.

A further aspect of the invention is a pharmaceutical composition for the treatment of pain comprising at least one GLP-1 agonist, and optionally a pharmaceutically acceptable carrier, diluent or/and auxiliary substance. Pain is the pain described herein. In particular, pain may be associated with (a) stenosis or/and occlusion in the biliary tract, or/and (b) biliary dyskinesia. Treatment of pain can be a palliative treatment, specifically a palliative treatment as described herein. The composition is a composition as described herein.

Another aspect of the invention is a method of treating a disease or condition associated with (a) stenosis or/and occlusion in the biliary tract, or/and (b) biliary dyskinesia, the method comprising comprising at least one GLP-1 The agonist, and optionally the pharmaceutical composition of the pharmaceutically acceptable carrier, diluent or/and auxiliary substance, is administered to the individual in need thereof. In the method of the invention, the disease or condition may be any of the diseases as described herein, in particular, diseases associated with (a) stenosis or/and occlusion in the biliary tract, or/and (b) biliary dyskinesia. . The composition is a composition as described herein.

A patient or/and an individual treated by a pharmaceutical composition or/and a method as described herein can be a mammal, including humans and non-human mammals. Preferred patients or/and individuals are humans.

Specifically, the patient does not have diabetes, such as type 1 or type 2 diabetes.

Specifically, the patient is not obese. More specifically, the patient's body mass index is less than 30 kg/m ² or less than 27 kg/m ² .

Specifically, the patient does not have a CNS disorder, such as Alzheimer's or Parkinson's.

A further aspect of the invention is the use of a GLP-1 agonist for the manufacture of a medicament for treating a disease or condition associated with (a) stenosis or/or occlusion in the biliary tract, or/and (b) biliary dyskinesia The medicament comprises at least one GLP-1 agonist, and optionally a pharmaceutically acceptable carrier, diluent or/and auxiliary substance. In the use of the invention, the disease or condition may be any of the diseases as described herein, in particular associated with (a) stenosis or/and occlusion in the biliary tract, or/and (b) biliary dyskinesia. disease. The drug is a composition as described herein.

In the present invention, the "anticonvulsant activity" or "antimony activity" of the GLP-1 agonist means the degree of tension of the smooth muscle, specifically, in the biliary system, which is lowered by administration of the GLP-1 agonist. Specifically, lixisenatide provides anticonvulsant or antispasmodic activity.

In the present invention, the at least one GLP-1 agonist may be one, two, three, four, five or more GLP-1 agonists. In particular, the at least one GLP-1 agonist can be a GLP-1 agonist.

In the present invention, the term "GLP-1 agonist" includes GLP-1, analogs and derivatives thereof, exenatide-3, analogs and derivatives thereof, and exenatide-4, which are similar Things and derivatives. Also included are substances that exhibit the biological activity of GLP-1. The "GLP-1 agonist" is also known as the "GLP-1 receptor agonist."

The pharmaceutical composition of the present invention may comprise one or more selected from the group consisting of: glucagon-like peptide-1 (GLP-1), GLP-1, independently of each other. Analogs and derivatives, Exendin-3, analogs and derivatives of Exendin-3, Exendin-4, Analogs and derivatives of Exendin-4.

GLP-1, Exendin-3 or/and Exendin-4, as used herein, includes pharmaceutically acceptable GLP-1, Exendin-3 or/and Exendin-4 salt.

The GLP-1 agonist is preferably lixisenatide or/and a pharmaceutically acceptable salt thereof.

The GLP-1 agonist is also preferably liraglutide or/and its pharmaceutically acceptable salt.

For example, GLP-1 analogs and derivatives are described in WO 98/08871; Exendin-3, analogs and derivatives of Exendin-3, and Exendin-4 and Exenatide Analogues and derivatives of -4 can be found in WO 01/04156, WO 98/30231, US 5, 424, 286, EP Application No. 99 610 043.4, WO 2004/005342, and WO 04/035623. These documents are incorporated herein by reference. Exendin-3 and Exendin-4 described in these documents, and the analogs and derivatives thereof described therein, can be used in the compositions of the present invention as GLP-1 agonists. Any desired combination of Exendin-3 and Exendin-4, and the analogs and derivatives thereof described in these documents, can also be used as the GLP-1 agonist.

Preferably, the at least one GLP-1 agonist is independently selected from the group consisting of analogs and derivatives of Exendin-4, Exendin-4, and pharmaceutically acceptable salts thereof.

A preferred GLP-1 agonist is an Exenatide-4 analog selected from the group consisting of: H-desPro ³⁶ - Exenatide-4-Lys ₆ -NH ₂ (desPro ³⁶ - Esser exenatide _{-4-Lys 6 -NH 2, AVE0010} , ^{lixisenatide), H-des (Pro 36,37} ) - exendin _{-4-Lys 4 -NH 2, H} -des (Pro 36,37 ) - Exenatide-4-Lys ₆ -NH ₂ , and a pharmacologically acceptable salt thereof.

A more preferred GLP-1 agonist is an Exendin-4 analog selected from the group consisting of: desPro ³⁶ [Asp ²⁸ ] Exendin-4 (1-39), desPro ³⁶ [IsoAsp ²⁸ Exenatide-4 (1-39), desPro ³⁶ [Met(O) ¹⁴ , Asp ²⁸ ] Exendin-4 (1-39), desPro ³⁶ [Met(O) ¹⁴ , IsoAsp ²⁸ ] Senamin-4 (1-39), desPro ³⁶ [Trp(O ₂ ) ²⁵ , Asp ²⁸ ] Exendin-2 (1-39), desPro ³⁶ [Trp(O ₂ ) ²⁵ , IsoAsp ²⁸ ] Senapeptide-2 (1-39), desPro ³⁶ [Met(O) ¹⁴ Trp(O ₂ ) ²⁵ , Asp ²⁸ ] Exendin-4 (1-39), desPro ³⁶ [Met(O) ¹⁴ Trp (O ₂ ) ²⁵ , IsoAsp ²⁸ ] Exenatide-4 (1-39), and a pharmacologically acceptable salt thereof.

A more preferred GLP-1 agonist is an Exendin-4 analog selected from the group described in the above paragraph, wherein the peptide-(Lys) ₆ -NH _{2 is} attached to the Exendin-4 analog C-side.

A more preferred GLP-1 agonist is an Exenatide-4 analog selected from the group consisting of: H-(Lys) ₆ -desPro ³⁶ [Asp ²⁸ ] Exendin-4 (1-39) )-Lys ₆ -NH ₂ , desAsp ²⁸ Pro ³⁶ , Pro ³⁷ , Pro ³⁸ Exenatide-4(1-39)-NH ₂ , H-(Lys) ₆ desPro ³⁶ , Pro ³⁷ , Pro ³⁸ [Asp ²⁸ Exenatide-4(1-39)-NH ₂ , H-Asn-(Glu) ₅ desPro ³⁶ , Pro ³⁷ , Pro ³⁸ [Asp ²⁸ ] Exenatide-4(1-39)-NH ₂ , desPro ³⁶ , Pro ³⁷ , Pro ³⁸ [Asp ²⁸ ] Exenatide-4(1-39)-(Lys) ₆ -NH ₂ , H-(Lys) ₆ -desPro ³⁶ ,Pro ³⁷ ,Pro ³⁸ [Asp ²⁸ ] Exendin-4 (1-39)-(Lys) ₆ -NH ₂ , H-Asn-(Glu) ₅ -desPro ³⁶ , Pro ³⁷ , Pro ³⁸ [Asp ²⁸ ] Exenatide-4 ( 1-39)-(Lys) ₆ -NH ₂ , H-(Lys) ₆ -desPro ³⁶ [Trp(O ₂ ) ²⁵ , Asp ²⁸ ] Exenatide-4(1-39)-Lys ₆ -NH ₂ , H-desAsp ²⁸ Pro ³⁶ , Pro ³⁷ , Pro ³⁸ [Trp(O ₂ ) ²⁵ ] Exenatide-4(1-39)-NH ₂ , H-(Lys) ₆ -desPro ³⁶ , Pro ³⁷ , Pro ³⁸ [Trp(O ₂ ) ²⁵ , Asp ²⁸ ] Exenatide-4(1-39)-NH ₂ , H-Asn-(Glu) ₅ -desPro ³⁶ , Pro ³⁷ , Pro ³⁸ [Trp(O ₂ ) ²⁵ , Asp ²⁸ ] Exenatide-4 (1-39-NH ₂ , desPro ³⁶ , Pro ³⁷ , Pro ³⁸ [Trp(O ₂ ) ²⁵ , Asp ²⁸ Exenatide-4(1-39)-(Lys) ₆ -NH2, H-(Lys) ₆ -desPro ³⁶ , Pro ³⁷ , Pro ³⁸ [Trp(O ₂ ) ²⁵ , Asp ²⁸ ] Exenatide -4(1-39)-(Lys) ₆ -NH ₂ , H-Asn-(Glu) ₅ -desPro ³⁶ , Pro ³⁷ , Pro ³⁸ [Trp(O ₂ ) ²⁵ , Asp ²⁸ ] Exenatide-4 (1-39)-(Lys) ₆ -NH ₂ , H-(Lys) ₆ -desPro ³⁶ [Met(O) ¹⁴ , Asp ²⁸ ] Exenatide-4(1-39)-Lys ₆ -NH ₂ , des Met(O) ¹⁴ Asp ²⁸ Pro ³⁶ , Pro ³⁷ , Pro ³⁸ Exenatide-4(1-39)-NH ₂ , H-(Lys) ₆ -desPro ³⁶ , Pro ³⁷ , Pro ³⁸ [Met( O) ¹⁴ , Asp ²⁸ ] Exenatide-4(1-39)-NH ₂ , H-Asn-(Glu) ₅ -desPro ³⁶ , Pro ³⁷ , Pro ³⁸ [Met(O) ¹⁴ , Asp ²⁸ ] Senamin-4(1-39)-NH ₂ , desPro ³⁶ , Pro ³⁷ , Pro ³⁸ [Met(O) ¹⁴ , Asp ²⁸ ] Exenatide-4(1-39)-(Lys) ₆ -NH ₂ , H-(Lys) ₆ -desPro ³⁶ , Pro ³⁷ , Pro ³⁸ [Met(O) ¹⁴ , Asp ²⁸ ] Exenatide-4(1-39)-Lys ₆ -NH ₂ , H-Asn-( Glu) ₅ desPro ³⁶ , Pro ³⁷ , Pro ³⁸ [Met(O) ¹⁴ , Asp ²⁸ ] Exenatide-4(1-39)-(Lys) ₆ -NH ₂ , H-(Lys) ₆ -desPro ³⁶ [Met(O) ¹⁴ , Trp(O ₂ ) ²⁵ , Asp ²⁸ ] Exenatide-4(1-39)-Lys ₆ -NH ₂ , desAsp ²⁸ Pro ³⁶ , Pro ³⁷ , Pro ³⁸ [Met(O) ^{_{14, Trp (O 2) 25}} ] exenatide _{-4 (1-39) -NH 2, H-} (Lys) 6 -desPro 36, Pro 37, Pro 38 [Met (O) 14, Trp (O 2) 25, Asp 28] exendin-4 ( 1-39)-NH ₂ , H-Asn-(Glu) ₅ desPro ³⁶ , Pro ³⁷ , Pro ³⁸ [Met(O) ¹⁴ , Asp ²⁸ ] Exenatide-4(1-39)-NH ₂ , desPro ³⁶ , Pro ³⁷ , Pro ³⁸ [Met(O) ¹⁴ , Trp(O ₂ ) ²⁵ , Asp ²⁸ ] Exenatide-4(1-39)-(Lys) ₆ -NH ₂ , H-(Lys) ₆ -desPro ³⁶ , Pro ³⁷ , Pro ³⁸ [Met(O) ¹⁴ , Trp(O ₂ ) ²⁵ , Asp ²⁸ ] Exenatide-4(1-39)-(Lys) ₆ -NH ₂ , H-Asn- (Glu) ₅ -despro ³⁶ , Pro ³⁷ , Pro ³⁸ [Met(O) ¹⁴ , Trp(O ₂ ) ²⁵ , Asp ²⁸ ] Exenatide-4(1-39)-(Lys) ₆ -NH ₂ , And pharmaceutically acceptable salts of such compounds.

A better GLP-1 agonist is selected from Arg ³⁴ , Lys ²⁶ (N ^ε (γ- glutamine sulfhydryl (N ^α - hexadecanyl))) GLP-1 (7-37) [Lila Rupin] and its pharmacologically tolerable salt composition.

A better GLP-1 agonist is lixisenatide. The lixisenatide has the sequence of H-desPro ³⁶ - Exenatide-4-Lys ₆ -NH ₂ (desPro ³⁶ Exenatide-4-Lys ₆ -NH ₂ , AVE0010). This is disclosed in SEQ ID NO: 93 in WO 01/04156. Lisila is a derivative of Exendin-4:

SEQ ID NO: 1: Lisila (44 AS)

HGEGTFTSDLSKQMEEEAVRL-FIEWLKNGGPSSGAPPSKKKK-KK-NH ₂

SEQ ID NO: 2: Exenatide-4 (39 AS)

HGEGTFTSDLSKQMEEEAVRL-FIEWLKNGGPSSGAPPPS-NH ₂

It is also preferred to provide a pharmaceutically acceptable salt of lixisenatide.

The term "at least one GLP-1 agonist" includes combinations of GLP-1 agonists for use in the compositions of the invention described herein, examples of which are two or more selected from the GLP-1 agonists described herein. Any combination of GLP-1 agonists required.

More preferably, the at least one GLP-1 agonist is independently selected from exenatide-4, H-desPro ³⁶ -exenatide-4-Lys ₆ -NH ₂ and Arg ³⁴ , Lys ²⁶ (N ^ε (γ) - glutamine-based (N ^α -hexadecanyl))) GLP-1 (7-37) [lilastuide] and a pharmacologically acceptable salt thereof.

The compositions of the present invention may contain a GLP-1 agonist at a dose of from 10 μg/ml to 20 mg/ml, preferably from 25 μg/ml to 15 mg/ml. For acidic to neutral soluble GLP-1 agonists, the dosage is preferably from 20 μg/ml to 300 μg/ml, and for neutral to alkaline agonists, they are preferably 500 μg/ Ml to 10 mg/ml. In the case of Exendin-4 analog, 20 μg/ml to 150 μg/ml is preferred.

In the present invention, the at least one GLP-1 agonist, in particular desPro ³⁶ - Exendin-4 (1-39)-Lys ₆ -NH ₂ or / and a pharmaceutically acceptable salt thereof, Dosages sufficient to elicit therapeutic utility are administered to an individual in need thereof.

In the present invention, the at least one GLP-1 agonist, in particular desPro ³⁶ - Exendin-4 (1-39)-Lys ₆ -NH ₂ or / and a pharmaceutically acceptable salt thereof, Formulated with a suitable pharmaceutically acceptable carrier, adjuvant or/and auxiliary substance.

The at least one GLP-1 agonist, in particular desPro ³⁶ - exenatide-4(1-39)-Lys ₆ -NH ₂ or / and a pharmaceutically acceptable salt thereof, can be administered orally For example, by injection (such as by intramuscular or by subcutaneous injection). Suitable injection devices are known, for example, so-called "pens" containing the active ingredient, as well as injection needles. The at least one GLP-1 agonist, in particular desPro ³⁶ - Exendin-4 (1-39)-Lys ₆ -NH ₂ or / and a pharmaceutically acceptable salt thereof, may be, for example, in a dose range A suitable dose of 10 to 15 μg per dose or 15 to 20 μg per dose is administered.

In the present invention, the at least one GLP-1 agonist, in particular desPro ³⁶ exenatide-4(1-39)-Lys ₆ -NH ₂ or/and a pharmaceutically acceptable salt thereof, may be present A daily dose ranging from 10 to 20 μg, ranging from 10 to 15 μg or ranging from 15 to 20 μg is administered. The at least one GLP-1 agonist, in particular desPro ³⁶ exenatide-4(1-39)-Lys ₆ -NH ₂ or/and its pharmaceutically acceptable salt, can be injected once a day Be cast.

In the present invention, the at least one GLP-1 agonist, in particular desPro ³⁶ exenatide-4(1-39)-Lys ₆ -NH ₂ or/and a pharmaceutically acceptable salt thereof, is provided It is a liquid composition. Those skilled in the art are aware of liquid compositions suitable for use in non-oral administration of lixisenatide. The liquid composition of the present invention may have an acidic or physiological pH. The acidic pH is preferably in the range of pH 1-6.8, pH 3.5-6.8 or pH 3.5-5. Physiological pH is preferably in the range of pH 2.5-8.5, pH 4.0-8.5 or pH 6.0-8.5. The pH can be adjusted by a pharmaceutically acceptable dilute acid (usually HCl) or a pharmaceutically acceptable dilute base (usually NaOH).

A liquid composition comprising the at least one GLP-1 agonist, in particular desPro ³⁶ Exenatide-4(1-39)-Lys ₆ -NH ₂ or/and a pharmaceutically acceptable salt thereof, may comprise Preservatives. Suitable preservatives can be selected from the group consisting of phenol, m-cresol, benzyl alcohol and p-hydroxybenzoate. A preferred preservative is m-cresol.

A liquid composition comprising the at least one GLP-1 agonist, in particular desPro ³⁶ exenatide-4(1-39)-Lys ₆ -NH ₂ or/and a pharmaceutically acceptable salt thereof, may comprise Tonicity agent. Suitable osmotic agents can be selected from the group consisting of glycerin, lactose, sorbitol, mannitol, glucose, NaCl, calcium-containing or magnesium-containing compounds such as CaCl ₂ . The concentrations of glycerol, lactose, sorbitol, mannitol and glucose are in the range of 100-250 mM. The NaCl concentration is at most 150 mM. A preferred osmotic agent is glycerin.

A liquid composition comprising the at least one GLP-1 agonist, in particular desPro ³⁶ exenatide-4(1-39)-Lys ₆ -NH ₂ or/and a pharmaceutically acceptable salt thereof, may comprise 0.5 The methionine of μg/mL to 20 μg/mL is preferably 1 μg/mL to 5 μg/mL. Preferably, the liquid composition contains L-methionine.

The invention will be further illustrated by the following figures and examples.

Example Summary

An embodiment of the invention is a randomized, double-blind, placebo-controlled, double-sequence, dual-treatment crossover study evaluating the efficacy of a single subcutaneous injection of 20 μg lixisenatide for gallbladder movement in healthy male and female subjects. Gallbladder movement has been analyzed by cholangiography. Cholecystokinin (CCK-8) has been administered 60 minutes after administration of a single dose of placebo or 20 μg of lixisenatide, followed by a single dose of ^99m Tc meclofenin (a ^99m Tc-labeled imine) Diacetic acid derivative). The procedure can be summarized as follows. By placing a radiation sensitive camera on the subject's abdomen, an "image" of the liver, bile duct and gallbladder is obtained, which corresponds to where the radioactive bile moves. The liver frame map was obtained at 1 frame/minute for 60 minutes after injection of lixisenatide or placebo with ^99m Tc injection. After a gallbladder view at 60 minutes, 0.02 μg/kg CCK-8 was administered via a continuous infusion pump for 60 minutes. The image was captured for at least another 60 minutes after the CCK-8 infusion.

In the placebo group, CCK-8 caused a decrease in the ^99m Tc count recorded by the gallbladder, indicating that bile was released from the gallbladder via the bile duct into the duodenum. Surprisingly, this effect of CCK-8 was greatly reduced by subcutaneous administration of a single dose of 20 μg lixisenatide in healthy subjects (see the exemplary filling curve and emptying curve in Figure 2). ). The ^99m Tc in the gallbladder is maintained at a high level, indicating that only a small portion of the bile is released. This utility has been quantified as follows. In the placebo group, CCK-8 caused a gallbladder excretion rate (GBEF, or excretion rate EF, which is the bile volume fraction excreted from the gallbladder) to increase by up to about 85%, as expected from the physiological effects of CCK. In the lixisenatide group, the excretion rate was unexpectedly maintained below 40% after CCK-8 administration (see the exemplary GBEF curve in Figure 3, the average is provided in Figures 7 and 9) . Subject to lixisenatide, 13/24 (54%) subjects had a GBEF of less than 40% at 60 minutes, compared to 1/24 (4%) of the placebo group. Subcutaneous administration of a single dose of 20 μg lixisenatide in healthy subjects significantly reduced gallbladder emptying (expressed as gallbladder excretion rate (GBEF)) caused by CCK-8 response compared to placebo at 60 45.8% in minutes.

Outline

1 list of abbreviations and terms

Ab antibody

AE adverse events

AEPM has pre-specified monitoring of adverse events

ALT alanine transaminase

ARAC Allergy Response Evaluation Committee

Area under the AUC concentration time curve

CCK cholecystokinin

CCK-8 cholecystokinin terminal amino acid fragment

CI confidence interval

Maximum plasma concentration observed by C _max

CRF case report

CT computed tomography

CV coefficient of variation

D days

DBP diastolic blood pressure

ECG ECG

FDA US Food and Drug Administration

GB gallbladder

GBEF gallbladder discharge rate

GLP-like glycosidin peptide

HCL hydrochloric acid

HIV human immunodeficiency virus

LLOQ lower limit of quantitation

MedDRA Standard Medical Dictionary for Pharmaceutical Administration

MRI magnetic resonance imaging

NaOH sodium hydroxide

P period

Potential clinically significant abnormalities in PCSA

QC quality control

QRS complex on QRS electrocardiogram (ventricular depolarization)

QTc QT interval for automatic correction of heart rate

QTcF QT interval, Fridericia correction

SAE critical adverse events

SBP systolic pressure

SD standard deviation

Standard error of SEM mean

SOC system organ class

^99m Tc 鎝99m

TEAE handles sudden adverse events

Time when t _max reaches C _max

Upper limit of the ULN standard

2 ethical considerations 2.1 Independent Ethics Committee or Audit Committee

Submit the proposal to an independent ethics committee and/or audit committee for review and written approval.

2.2 Ethical behavior of research

The programme complies with the recommendations of the 18th World Health Congress (Helsinki, 1964) and all applicable amendments. The program also complies with the laws and regulations of the country in which the study is conducted (UK) and any applicable guidelines.

2.3 Subject information and consent form

A test consent is obtained prior to any research-related steps. Subject test consent is modified in accordance with local regulations and requirements.

3 Introduction

Lixilata is an exenatide analog of the glucagon-like peptide 1 (GLP-1) receptor agonist developed for the treatment of type 2 diabetes.

There have been reports of cases of acute pancreatitis in patients treated with GLP-1 agonists, and the cause is still unknown. Whether pancreatitis is a type of potential utility or over-emphasis for GLP1 treatment is still under discussion. One hypothesis is that GLP1 agonists may alter the movement of the sphincter of the sphincter due to gastric bloating, causing the patient to have gallbladder (GB) mud or gallstone formation and thus a tendency to develop pancreatitis.

Cholecystokinin-excited cholangiography has been first described more than 30 years ago and is commonly used to calculate gallbladder excretion rate (GBEF) in bile mechanics and gallbladder movement. Biliary scintigraphy was performed after administration of a 99m ( ^99m Tc) labeled imine diacetic acid analog. These compounds have high affinity for liver uptake and are immediately secreted into the biliary tract and concentrated in the gallbladder. Therefore, a fat meal or exogenous cholecystokinin is used as a stimulator to induce gallbladder emptying. Low GBEF has been identified as a dysfunction of gallbladder dysfunction in patients with primary gallbladder dysfunction and Oddi's sphincter obstruction in the absence of calculi. Because GBEF is a variable that varies with the dose and duration of cholecystokinin 8 (CCK8, the terminal cystic acid fragment of CCK), many different values have been used to define gallbladder dysfunction; for accurate quantification of GBEF More and more evidence suggests that longer infusions (30 to 60 minutes) are better than shorter infusions (1 to 3 minutes).

This study was designed to evaluate the utility of a single subcutaneous injection of lixisenatide for stimulating gallbladder emptying (as an indirect measure of the possible effects on the eddy sphincter), expressed as GBEF caused by CCK8.

4 Research purposes the main purpose

‧ To assess the efficacy of 20 μg lixisenatide for gallbladder emptying expressed as gallbladder (GB) excretion rate (GBEF) due to continuous infusion of 0.02 μg/kg.h cholecystokinin (CCK8) at 60 minutes.

Secondary purpose

‧ Evaluate GBEF at 30 minutes.

‧ To assess the pharmacokinetic profile of a single subcutaneous injection of 20 μg lixisenatide.

5 research projects 5.1 Description of the overall research design and plan

This was a single-center, double-blind, randomized, placebo-controlled, single-dose, dual-dose, double-sequence, double-treatment crossover study comparing lixisenatide and saline injections in healthy male and female subjects (soothing) Agent).

Subjects were allowed to enter the facility on the morning of the first day. On the first day, after fasting overnight but free to drink, they were ready for cholangiography. Subjects were randomized to receive subcutaneous administration of 20 μg lixisenatide or placebo followed by rapid intravenous infusion of radiolabel. After the injection of ^99m Tc, the image was taken for 60 minutes. After viewing the gallbladder at 60 minutes, CCK8 was infused over 60 minutes; GB images were obtained for at least another 60 minutes, followed by CCK8 infusion.

Blood samples for determination of lixisenatide concentrations were obtained up to 12 hours after lixisenatide injection.

Subjects were randomly assigned to either Sequence 1 or Sequence 2 in a double-blind manner (Figure 1):

Sequence 1

‧ Phase 1: Placebo

‧No. 2: 20 μg Lixiula

Sequence 2

‧ Phase 1: 20 μg Lixiula

‧ Phase 2: Placebo

The total duration of each subject's participation in the study is 7 to 42 days and consists of the following:

‧Screening: 2 to 28 days

‧ Processing period 1: 2 days (Day 1, Day 1) includes 1 processing day

‧ clearance: at least 48 hours between doses, up to 7 days (between the first day of the first phase and the first day of the second phase)

‧ Processing period 2: 2 days (Day 1 to Day 1) includes 1 processing day

‧End of study: 2 to 7 days after the last dose

No analysis of inferiority analysis. The program has not been modified.

The interview schedule is described in Table 1.

5.2 Study design and discussion of selection of control groups

This randomized, placebo-controlled, double-sequence, double-treatment crossover study was designed to evaluate the utility of a single dose of lixisenatide for gallbladder emptying. Crossover design was chosen to avoid the effects of variability between subjects.

The study included male and female subjects who matched the target population of type 2 diabetes. Although most patients with Odysium sphincter dysfunction and recurrence of acute pancreatitis have been found to be female (1), there is no difference in GBEF between men and women in healthy subjects. Only post-menopausal women are included to avoid exposing unwanted radiation to women who are at risk of becoming pregnant. Although significant impairment of GB emptying has been reported in obese patients with type 2 diabetes (2), no statistically significant correlation between body mass index and GBEF was observed in healthy subjects. Therefore, male and female subjects with a body mass index of up to 35 kg/m ² were included in the study.

In order to allow bile to enter the gallbladder as much as possible by maximal relaxation of the gallbladder and maximum contraction of the Odyssey sphincter (3), biliary scintigraphy was performed in the fasted state. The radiolabel was injected directly after administration because the lixisenatide _tmax was about 1.5 to 2 hours.

When the absorption is complete (average peak plasma concentration of about 100 pg/mL at about 1.5 hours after injection), lixisenatide disappears rapidly from the blood circulation so that the short-clearing period can be 2 days, and at the last administration The study ended the visit within 1 week.

Because lixisenatide is a potential peptide that causes allergic reactions, this study hired an allergic response assessment committee to assess allergic or allergic reactions that occurred during the study.

5.3 Selection of research groups

A total of 24 subjects were recruited to have 20 subjects to be assessed (the gender of each randomized group was at least 30%).

Subjects were included in the study according to the following criteria.

5.3.1 Inclusion criteria demography

I1. Inclusion of male or female subjects between the ages of 35 and 65.

I2. The included body mass index ranged from 18.0 to 35.0 kg/m ² .

health status

I3. Confirmed to be healthy according to a full clinical assessment (detailed medical history and complete physical examination).

I4. Normal signs of vitality after 10 minutes of rest in the supine position: -95 mmHg<systolic pressure <160 mmHg -45 mmHg<diastolic pressure<90 mmHg -40 bpm<heart rate <100 bpm

I5. The standard 12-lead electrocardiogram (ECG) is normal after 10 minutes of rest position; 120 ms<PR<220 ms, QRS<120 ms, QTc 430 ms (if male), QTc 450 ms (if female)

I6. Laboratory parameters fall within the normal range unless the investigator believes that the abnormality is clinically irrelevant for healthy subjects; but serum creatinine, alkaline phosphatase, liver enzyme (aspartate transaminase) , alanine transaminase, bilirubin) does not exceed the upper limit of the laboratory's normal value (unless the subject's written record is Gilbert's syndrome)

I7. If female abortion persists for more than 3 months or after menopause; menopause is defined as over 60 years of age, or between 45 and 60 years of amenorrhea for at least 2 years, plasma follicle stimulating hormone concentration >30 UI/L. It was confirmed to be healthy according to a full clinical evaluation (detailed medical history and complete physical examination).

Regulation

I8. A written test consent must be provided prior to any procedures related to the study.

19. Protected by the applicable health care system and/or by the recommendations of the national law applicable to relevant biomedical research

I10. Not subject to administrative or legal supervision

5.3.2 Exclusion criteria Medical history and clinical status

E1. Clinically any history or presence of cardiovascular, pulmonary, gastrointestinal, hepatic, renal, metabolic, hematological, neurological, musculoskeletal, joint, mental, systemic, ocular, gynecological (if female) or infectious diseases Or signs of acute disease

E2. Frequent headaches and / or migraine, recurrent nausea and / or vomiting (more than 2 times a month)

E3. Donate blood within the first 3 months of inclusion, no matter how much volume

E4. Symptomatic hypotension (all blood pressure reduction) or non-symptomatic hypotension, as defined by a decrease in systolic blood pressure ≧ 20 mmHg when changing from supine to standing position within 3 minutes

E5. The presence or history of a drug-allergic or clinically significant allergic disease diagnosed and treated by a clinician

E6. History or presence of drug or alcohol abuse (weekly alcoholic drinking male > 21 units and female > 14 units [1 unit = 1/2 pint of beer, 25 mL of 40% alcohol or 125 mL red wine cup] )

E7. Draw more than 5 cigarettes per day or equivalent, unable to quit during the study period

E8. Excessive consumption of beverages containing xanthine base (>4 cups or 4 bottles per day)

Interfering substance

E9. Any drug (including St. John's) within 14 days prior to inclusion Grass), or reduce the half-life or half-life of the drug within 5 times, vaccinate any vaccine within 28 days

General condition

E10. At the discretion of the investigator, any subject who may not be able to cooperate during the study or who is unable to cooperate because of language problems or poor mental development

E11. Any subject who was in the exclusion period of the previous study in accordance with the implementation regulations

E12. Any subject who cannot be contacted in an emergency

E13. Any subject directly involved in the programme or any associate researcher, research assistant, pharmacist, research coordinator, or other staff member

Biological state

E14. Positive for any of the following tests: hepatitis B surface antigen, anti-hepatitis C virus antibody, anti-human immunodeficiency virus 1 and 2 antibodies (anti-HIV1 and anti-HIV2 Ab)

E15. Urine drug screening is a positive result (amphetamine / methamphetamine, barbiturate, benzodiazepine, marijuana, cocaine, opiates)

E16. Alcohol test positive

Research characteristics

E17. Acute diarrhea or constipation within 7 days prior to the first study day. If the screening occurs >7 days before the first study day, the criteria are determined on the first study day. Diarrhea is defined as the passage of liquid feces and/or the frequency of bowel movements exceeds 3 per day. Times. Constipation is defined as the inability to defecate more than every other day.

E18. Radiation exposure from clinical trials, including from the investigator, excludes background radiation but includes diagnostic X-rays and other medical exposures, exceeding 5 mSv in the last 12 months or more than 10 mSv in the last 5 years. Workers without occupational exposure, as defined by the Ionizing Radiation Regulations 1999, are permitted to participate in the study.

E19. Lixisenatide previously used, exenatide (Byetta ^TM) or other non-oral GLP1 agonist

E20. History of pancreatitis, chronic pancreatitis, gallbladder disease, cholecystectomy, stomach/stomach surgery, and inflammatory bowel disease

E21. There is gallstone or clinically significant liver abnormality based on ultrasound scanning

5.3.3 Subject leaves therapy or assessment

If the subject decides to withdraw from the treatment at any time and regardless of the reason, they can do so, or the subject can withdraw because of the researcher's decision.

Subjects who are diagnosed with an allergic reaction and who are considered by the researcher to be extremely relevant to the study product are subject to additional treatment.

Specific suspension requirements are considered from screening to the end of the study. The study product was discontinued in the case of manual reading of QTcF (QTc, Federian correction) extension (automatic measurement: ≧ 500 ms) and in the case of pregnancy. In addition, in any case of diagnosis of acute pancreatitis, the use of the study will be discontinued with other suspected drugs and clinically targeted to the subject. Follow-up visits.

5.4 Processing 5.4.1 Research products 5.4.1.1 Lisila

‧Pharmaceutical form: a sterile aqueous solution in 3-mL glass vial for subcutaneous injection containing 300 μg of active ingredient (ie 100 μg/mL), glycerol, sodium acetate trihydrate, methionine, m-cresol, HCL/NaOH and water for injection.

‧ Routes and methods of administration: Subcutaneous administration using a pen-type syringe (OptiClik®).

‧ Drug dosage per administration: According to the random grouping schedule, 20 μg lixisenatide is given once in the morning of the first or second day of the second phase.

• The study product was administered by deep subcutaneous injection in the left or right anterior lateral abdominal wall.

‧ For proper dosing, the OptiClik pen unit is set as follows: 20 μg lixisenatide = 20 units (= 200 μL) on OptiClik.

‧ Lisila is provided by the sponsor.

5.4.1.2 Placebo

‧ Pharmaceutical form: A sterile aqueous solution in 3-mL glass vials is administered for subcutaneous injection containing sodium chloride, m-cresol, HCL/NaOH and water for injection.

‧ Routes and methods of administration: Subcutaneous administration using a pen-type syringe (OptiClik).

‧Dose dose per dose: According to the randomization schedule, 200 μL of placebo was given once in the morning of the first or second day of the second period.

• The study product was administered by deep subcutaneous injection in the left or right anterior lateral abdominal wall.

• For proper dosing, the OptiClik pen unit is set as follows: 200 μL placebo = 20 units on OptiClik.

‧ Placebo is provided by the sponsor.

The OptiClik pen injector is provided by the sponsor to each subject for injection of the research product. Needle ^{(Ypsomed Optifine TM 8,8 mm X 31G} , order number 3100564) was purchased by the clinical setting. Disposable materials are kept by the researcher until the end of the study, and the sponsors carry out all documentary reconciliation.

5.4.1.3 Non-research products 5.4.1.3.1 Radiopharmaceutical ( ^99m Tc Meclofenin/Cholediam)

The Cholediam® kit for the preparation of the ^99m Tc mebromophenin injection was supplied by Quotient Clinical and contained 40 mg of mebronfinin and 0.6 mg of anhydrous stannous chloride. The effective dose in this study did not exceed 60 MBq (1.44 mSv) per period and did not exceed 120 MBq (2.88 mSv) in total.

5.4.1.3.2 Cholecystokinin 8 (Kinevac / Xinkalite)

Cholecystokinin 8 (CCK8) is supplied by the sponsor with Xinkalite for injection (Kinevac®). The vial containing 5 μg of simikanit was reduced using 5 mL of sterile water. Next, 2.5 μg of cinkarite was diluted to 50 mL in a 50 mL syringe using saline and infused according to the following formula: infusion rate (mL/hr) = body weight (kg) x 0.02 (CCK8 dose) x 20 (containing 1 Gg CCK8 solution volume)

5.4.2 Studying the characteristics of the product 5.4.2.1 Research products

The study product was supplied by a sponsor in a sterile aqueous solution in 3-mL glass vial for subcutaneous injection containing 300 μg/mL lixisenatide (100 μg/mL), glycerol, sodium acetate trihydrate, methionine , m-cresol, HCL/NaOH and water for injection (batch number FRA01282/40C008/C1005517).

Placebo is provided as a sterile aqueous solution for subcutaneous injection containing sodium chloride, m-cresol, HCl/NaOH and water for injection (batch number FRA01419/40C006/C1005518).

5.4.2.2 Non-research products

鎝99m ( ^99m Tc) mebromophenone is supplied by Quotient Clinical for injection kits in Cholediam; each tube contains 40 mg of mebronfinin and 0.6 mg under sterile nitrogen in a sterile, pyrogen-free, lyophilized solution. Anhydrous stannous chloride (batch number FRA01419/40C006/C1005518).

Cholecystokinin-8 is supplied by the sponsor with Xinkalite for injection (Kinevac) and contains 5 μg of Xinkalite (batch C1008567) in the small tube.

5.4.3 Method of assigning subjects to treatment groups

The randomized processing set number table is mainly generated by sanofiaventis.

Subjects who met all inclusion/exclusion criteria were assigned before the study product was placed on the first day of the first study period:

‧Based on the inclusion of subject numbers with increasing chronological order.

‧The processing number is in the order of the pre-plan according to the subject number (corresponds to the number of the processing order). The processing number emulates the randomized processing set number table. Subjects received study medications based on their randomized treatment set number.

Subjects were randomized into either Sequence 1 or Sequence 2. The random grouping ratio was 1:1 and stratified by gender to ensure that at least 30% of each gender was treated in the study. See the research protocol for more details.

5.4.4 Dose selection in the study

Based on the Phase 2 dose range study, the 20 μg dose of lixisenatide used in this study was considered to provide the best benefit-risk ratio and was well tolerated in both non-diabetic and diabetic populations.

The risk associated with the maximum possible dose of radiation used is very small and considered acceptable. The effective ^99m Tc dose received by each subject did not exceed 60 MBq (1.44 mSv) for one administration and 120 MBq (2.88 mSv) for two administrations. This is in line with the management of the Radioactive Substance Advisory Committee, which recommends that the ^99m Tc dose for the gallbladder diagnostic procedure does not exceed 150 MBq and is only slightly higher than the average natural background radiation dose (2.7 mSv) accepted in the UK each year.

Xinkalite is the only CCK8 analogue approved by the US Food and Drug Administration (FDA). Although in the published study, CCD8 dose can be 0.01 to 0.5 μg/kg and rapid injection infusion duration is 1 to 60 minutes, short infusion causes abdominal cramps and nausea, and makes the normal injection range of GB injection rate It is difficult to achieve (4). In a study of 3 different CCK8 infusion methods, a dose of 0.02 μg/kg was administered to 60 subjects at 15 minutes, 30 minutes, and 60 minutes of infusion, showing a 60-minute infusion compared to healthy subjects. In 15 minutes and 30 The shorter infusion time in minutes has the lowest variability (5). Therefore, in the present study, an infusion of 0.02 μg/kg lasted for 60 minutes.

5.4.5 Screening and scheduling of doses for each subject 5.4.5.1 Research products 5.4.5.1.1 Lisila

A 20 μg dose of lixisenatide was administered once in the fasted state on the morning of day 1 of the first or second phase, according to a randomized schedule. A 20 μg dose of lixisenatide corresponds to 20 units (= 200 μL) on the OptiClik pen.

5.4.5.1.2 Placebo

Placebo (200 μL) was administered once in the fasted state on the first morning of the first or second phase according to a random grouping schedule (20 units on the OptiClik pen).

5.4.5.2 Non-research products 5.4.5.2.1 Radiopharmaceutical ( ^99m Tc Meclofenin/Cholediam)

Immediately after the administration of lixisenatide or placebo, a single dose of ^99m Tc meclofenin was intravenously infused on the first morning of each phase after at least 10 hours of overnight fasting.

5.4.5.2.2 Cholecystokinin 8 (Kinevac / Xinkalite)

A single dose of CCK8 (0.02 μg/kg) was infused 60 minutes after the radioactive label was administered on the first day of each period and 60 minutes later.

5.4.6 Blind method

Except for bioanalysts and pharmacokinetics responsible for sample analysis and pharmacokinetic assessment, all involved in the study were blinded until the database was locked.

The study product was indistinguishable from the placebo system and the injection volume was the same. The processing kits and corresponding medicines are marked with the number generated by the Sanofi Avantis computer program. Researchers cannot obtain randomization (processing) coding unless the knowledge of the research product is necessary to process the subject.

Review the ARAC number and determine the allergic or allergic reaction in a blinded manner.

5.4.7 Previous or combined therapy

The treatment of drugs that could not be used prior to inclusion is indicated in paragraph 5.3.2.

Combined drug treatment was not allowed during the study. However, if a particular treatment is necessary for any reason, an accurate record must be maintained on the appropriate record, including the name of the medical treatment (international non-proprietary name), the daily dose, and the duration of use.

5.4.8 Dealing with compliance

Subject to research by direct medical supervision. The actual dose and actual time of each dose are recorded in the case report. A one month check was performed to check the acceptance of the study product based on the subject.

5.5 Pharmacodynamics, safety and pharmacokinetic assessment

A summary of safety, pharmacokinetic, and pharmacodynamic assessments is presented in Table 1; detailed timelines for each period are provided in Table 2.

5.5.1 Pharmacodynamic evaluation

Gallbladder emptying after stimulation by CCK8 is expressed as GBEF, a percentage change in net GB count after administration of the stimulator. The GBEF was evaluated according to the following formula at 30 minutes and 60 minutes after the start of CCK8 administration:

5.5.1.1 Pharmacodynamic measurement and time selection

After overnight fasting, subjects received 20 μg of lixisenatide subcutaneously and immediately received an intravenous infusion of ^99m Tc-labeled mebromophenine, while lying upside down under a large-field gamma camera. Liver phase images were taken at 1 frame per minute for 60 minutes. After GB was visualized at 60 minutes, 0.02 μg/kg CCK8 was administered via a constant infusion pump for 60 minutes. Continue to capture images for 1 minute at the beginning of the CCK8 infusion for at least another 60 minutes until the CCK8 infusion is complete. A background-corrected time-activity curve is obtained by dragging the region of interest to the periphery of the gallbladder and the background (near the normal liver) according to the frame showing the clear image of the gallbladder. Because the duration of the evaluation period was short, no correction was made for radioactivity decay.

5.5.1.2 Main pharmacodynamic variables

The main pharmacodynamic variables were GBEF at 60 minutes after the start of CCK8 infusion on Day 1 of Phase 1 and Day 2 of Phase 2.

5.5.1.3 Secondary pharmacodynamic variables

The secondary pharmacodynamic variables were GBEF at 30 minutes after the start of CCK8 infusion on Day 1 of Day 1 and Day 2 of Phase 2. During the study period GBEF is also collected every 2 minutes.

5.5.2 Security variables and time selection for evaluation

Security is monitored via the following: ‧ spontaneous reporting of adverse events observed by the subject or researcher; ‧ Clinical laboratory tests (biochemistry, hematology, urine tests); ‧ Electrocardiogram (ECG) records (heart rate; PR, QRS, QT interval; QTc) ‧Measurement of vital signs (heart rate, systolic blood pressure and diastolic blood pressure; weight, height [used to calculate body mass index], body temperature); ‧Body checkup

Monitor clinically significant abnormalities, if any, until recovery or until clinically stable.

Serology (hepatitis B surface antigen, hepatitis C antibody, anti-HIV1 and anti-HIV2 antibodies) was performed only at the time of screening. Urine drug screening and alcohol breath tests were performed on the first day of screening and each period.

5.5.2.1 Adverse events

From signing the trial consent to the end of the study visit, record all adverse events, no seriousness or association with the research product. For each adverse event, the investigator should specify the date of occurrence, intensity, behavior to the study product, the improved treatment provided, and the outcome, and provide a reasonable likelihood of assessing whether the adverse event is related to the study product.

Critical Adverse Events (SAE) are any unsuitable medical events that occur at any dose: ‧ cause death; or ‧ life-threatening; or ‧ need to be hospitalized or extended to continue hospitalization; or ‧ cause persistent or apparent loss of power / disability; or ‧ congenital anomalies / birth defects; or ‧ medically important events

5.5.2.1.1 Adverse events with specific specified monitoring in advance

Adverse events that need to be specified in advance (AEPM) are defined as adverse events (critical or non-critical) that are monitored, documented, and processed in a manner specified in advance as specified in the protocol.

If you have any of the following, please notify the sponsor immediately:

‧ALT increases ≧2x standard value upper limit (ULN)

‧QTc≧500 ms

‧pregnancy

‧ Symptomatic dose is too high

‧ Pancreatitis and / or pancreatic enzymes (amylase, lipase) increase > 2x ULN AEPM without immediate notice is:

‧non-symptomatic dose is too high

‧Local tolerance

‧ If the scoring system described in Annex C of the programme is used, the skin response of ≧1 is scored. The protocol specifying the score ≧3 is reported as an adverse event, and if the stimuli score is 3 or 4, the dermatologist should be consulted. This information was corrected in the file annotation (November 2, 2009) to report the skin reaction of the score ≧1 and scored ≧4 to consult a dermatologist.

‧ Allergic reactions or allergic reactions.

5.5.2.2 Laboratory safety parameters

Standard clinical laboratory parameters (biochemistry, hematology, urine tests) were measured at screening, on day 1 of each phase, and at the end of the study (Table 1). Other tests may be performed during the study according to the medical judgment of the investigator. Blood samples were taken in the fasted state.

‧Biochemistry

- Plasma / serum electrolytes: sodium, potassium, calcium, chloride

- Liver function: ALT, aspartate transaminase, γ-glutamine invertase, alkaline phosphatase, total bilirubin and conjugated bilirubin

- Metabolism: total cholesterol, triglycerides, glucose, albumin, total protein

- Potential muscle toxicity: creatinine phosphokinase

- Renal function: urea, creatinine

- Pancreatic function: amylase, lipase

‧hematology

- red blood cell count, blood volume ratio, heme and platelets

- Differences (neutrophils, lymphocytes, basophilic balls, mononuclear balls and eosinophils) white blood cell count

- International normalized ratio and activated partial thrombin time

‧ urine test: measuring rod protein, glucose, red blood cells, white blood cells (if positive for quantitative test), ketone body, pH

Pre-specify laboratory safety parameters for monitoring

Monitor, document and handle the following laboratory anomalies.

‧ Neutrophic neutropenia

‧Thrombocytopenia

‧ acute renal insufficiency

‧ suspected rhabdomyolysis

5.5.2.3 Other security parameters Sign of life

Heart rate, blood pressure (systolic and diastolic blood pressure measurements), and body temperature were measured at screening, on the first day of each phase, and at the end of the study visit. Heart rate and blood pressure were obtained after 10 minutes in the standing position and 3 minutes in the standing position.

Electrocardiogram

A standard 12-lead ECG (recorded at 10 mm at 25 mm/s, 10 mm/mV) was recorded after at least 10 minutes of elevation. The ECG parameters derived from automatic measurements are HR, PR, QRS, QT, and QTc.

Body checkup

Physical examination includes cardiac and respiratory auscultation; peripheral arterial pulse; pupil, knee, Achilles, and Babinski reflex; peripheral lymph node examination; and abdominal examination.

5.5.3 Pharmacokinetic assessment and time selection 5.5.3.1 Pharmacokinetic measurement and time selection

The sampling time for blood collection can be found in the schedule (Table 2).

A bioassay for measuring lixisenatide in plasma samples is summarized in Table 3.

All plasma samples from subjects treated with lixisenatide were analyzed. The bioanalyst analyzed the samples taken by the placebo recipients only 1.5 hours after dosing (P03), which bioanalysts were not too early blind.

5.5.3.2 Pharmacokinetic variables

Table 4 lists the main pharmacokinetic parameters, which are determined based on the plasma concentration of lixisenatide. Part of the AUC during the emptying of the gallbladder was added to analyze the individual effects of the exposure.

5.5.4 Measuring fitness

Standard measurements suitable for evaluation purposes were used in this study. The primary assessment index, GBEF, is commonly used to assess gallbladder emptying, while cholecystokinin-induced cholangiography is used in the study of biliary mechanics and gallbladder movement. It is used to calculate GBEF.

5.6 Data Quality Assurance

Fixed site monitoring ensures the quality of the test. The processing of clinical trial data is based on the following regulations and procedures. Data entry, verification, and validation were performed using standard computer software (Oracle® Clinical version 4.5.1); data was stored on a digital VMS computer in an Oracle database. The double-entry method is used to ensure that data (except for comments) is accurately transferred from the CRF to the database. In addition, each database change can be tracked using an audit trail. Develop a data check method to limit all automatic validity checks and additional manual checks to ensure data quality. Explore all the differences until they are resolved.

Sanofi Avantis held a meeting of researchers and began a meeting at a number of locations to gradually develop a common understanding of clinical research protocols, case reports, and research procedures.

Pharmacokinetic data manipulation and data quality assurance

Demographic data, date and time of dosing, date and time of sampling, and concentration were electronically transmitted to the pharmacology database of the Oracle Clinical and Watson database. Quality control (QC) checks the concentration data transmitted to Watson; no difference is detected. The pharmacokinetic parameters provided in all tables are computer generated. When calculating the average value of the concentration, the concentration value below the lower limit of quantitation (LLOQ) is regarded as 0; in terms of calculating the pharmacokinetic parameters, if it appears before C _max is regarded as 0, otherwise it is "missing".

The mean calculation and its associated statistics are generated from values that are not rounded and may be slightly different from those measured using rounded values. For the final pharmacokinetic analysis, the drug will be administered The parameters were transmitted electronically to the Department of Biostatistics for further statistics. In all report forms, the concentration and pharmacokinetic parameter values are rounded off to 3 significant digits.

All raw data for the bioassay and pharmacokinetic segments of this study were kept in the appropriate file.

Examples of gallbladder filling and emptying curves and GBEF (%) in normal healthy subjects receiving placebo and lixisenatide are provided (Figures 2 and 3).

5.7 Statistical methods for planning and determining sample size 5.7.1 Statistics and analysis plans

The details of the statistical methods are summarized below. Clinical data analysis was performed by Sanofi Avantis Biostatistics and Programs using SAS® (SAS/Unix V9.2, SAS Institute, NC USA). Statistical analysis of pharmacokinetic parameters was performed by the Drug Handling Department using the Pharmacokinetic Data Management System (PKDMS) (Internal Software Version 2.0 and WinNonlin Professional® Version 5.2.1).

5.7.1.1 Analysis of pharmacodynamic variables

All pharmacodynamic analyses were performed using pharmacodynamic populations.

5.7.1.1.1 Description of pharmacodynamic variables

The pharmacodynamic evaluation index was GBEF caused by continuous infusion of 0.02 μg/kg CCK8 as measured by cholelithiasis at 60 minutes (major variable) and 30 minutes (minor variable). GBEF, provided by Quotient Clinical, is equivalent to a percentage change in the net GB count after administration of the stimulant.

5.7.1.1.2 Main analysis

The GBEF at 60 minutes was analyzed using a linear mixed utility model.

GBEF _{at 60 minutes} = order + period + gender + processing + error

Among them, fixed gender, order ("lixiela-placebo"-pair" placebo-lixilais), period (1 to 2) and treatment (lixiela vs placebo), and treatment The SAS PROC MIXED was used for the non-structural R matrix (i, j), the number of variances and the covariation of the subjects between the sequences.

Differences between treatment groups and corresponding 95% CI were estimated within the framework of a linear mixed utility model. If the upper limit of 95% CI on both sides of the GBEF absolute difference (placebo minus Lisila) between the two treatment groups was less than 0.20 (or 20%), then the inferiority was confirmed.

It is graphically explored whether the GBEF at 60 minutes is a normal distribution. If there is a significant deviation from the normal distribution, the plan uses the no-parent method.

5.7.1.1.3 Secondary analysis / secondary variable analysis Analysis of secondary assessment indicators

For the GBEF at 30 minutes, the same linear mixed-use utility model was performed as described above to estimate the difference between treatment groups and the corresponding 95% CI. Similarly, if you need to plan, use the no-parent method.

Narrative statistics and graphs

The GBEF at 30 minutes and 60 minutes was grouped according to the treatment group and listed by subject, sequence and interview. Provide the same narrative statistics by gender.

The box-and-whisker charts for the two assessment indicators are provided in accordance with the treatment group; individual maps are also provided.

In addition, GBEF data were drawn separately from the subjects every 2 minutes after the start of CCK8 infusion, summarized by median and averaged and ranked according to treatment groups.

Variance component

For the 30 minutes and 60 minutes of GBEF, intra-subject, inter-subject, and total standard deviation were estimated by observing the observed and expected mean squares within the framework of the following linear mixed utility model: GBEF _{At 30 or 60 minutes} = order + period + gender + treatment + subject (sequence) + error

Among them were fixed gender, order ("lixiela-placebo"-pair" placebo-lixilais), period (1 to 2), and treatment (lixiela vs. placebo), and between The random effects of the subjects used SAS PROC MIXED. 90% CI was calculated using the simple χ ² method (in the subject SD) and the Graybill Wang program (total SD) (6).

List

The list of GB biliary scintigraphy data (GBEF values at 30 minutes and 60 minutes, dates and times of biliary scintigraphy) is provided by subject and order.

5.7.1.2 Analysis of security data

Safety assessment is based on examining individual values (clinical significant abnormalities), narrative statistics (induction tables, graphs), and, if necessary, statistical analysis (appropriate estimates, confidence intervals). Safety analysis was performed based on the Sanofi Avantis document "induction and reporting of clinical pharmacology test data".

All security analyses are performed using a security community.

For all safety data, the observation period is divided into 3 sections:

‧ The pre-treatment period is defined as the time when the subject provides the trial consent and the first time the study product is administered in the first phase (excluded).

‧ For each treatment period, the processing period is defined as the time from the investment of the research product to the time of the investment of the research product.

‧ The post-processing period is defined as the time from the processing period to the next period (for the first period) to the study product or the end of the tracking period (for the second period).

All safety analysis is based on the processing period.

The list of potential clinically significant abnormalities (PCSA) used in the statistical analysis of laboratory parameters, vital signs, and ECG data is defined as version 2.0, dated September 14, 2009.

5.7.1.2.1 Adverse events 5.7.1.2.1.1 Definition

Adverse events were coded according to the standard medical dictionary of pharmacy management (MedDRA version 13.0). They are classified into pre-defined standard classes based on timing criteria:

‧Adverse events before treatment: Adverse events that develop or worsen in the early stages of treatment

‧ Handling of Unexpected Adverse Events (TEAE): Adverse events that occur or worsen during the processing period.

‧Adverse events after treatment: Adverse events that occurred later in the treatment.

TEAE is assigned to the accepted research product (lixila or placebo) at the time of the adverse event. If the TEAE develops at one processing and deteriorates at a later processing, it is considered that both processing are processing bursts.

If the start date (or time) of an adverse event is incomplete or missing, the adverse event is considered to be a TEAE in each period, unless part of the date (or time) or comment appears as a pre- or post-treatment event.

All adverse events reported in the study were listed on an individual basis, with flags indicating adverse event status. This list is sorted by subject, treatment, start date and time. However, the analysis of adverse events focuses on TEAE.

5.7.1.2.1.2 Handling unexpected incidents

The number and percentage of subjects with any TEAE, any critical TEAE, any severe TEAE, any TEAE that caused a permanent treatment discontinuation, or any TEAE that caused death (only when it occurred) were summarized by treatment group.

Subjects who presented TEAE were sorted by treatment group, major system organ class (SOC, classified according to MedDRA level), and preferred language (PT).

The TEAEs are summarized according to the treatment group. The list shows: ‧ the number and percentage of subjects with at least 1 TEAE in each SOC and total SOC; ‧ the number and percentage of subjects experiencing the preferred language in each SOC; The number of occurrences of all preferred words in each SOC and total SOC; ‧ the number of occurrences of each preferred language in each SOC.

5.7.1.2.1.3. Death, crisis and other significant adverse events

List any deaths, SAEs, or other significant adverse events by subject, date and time of onset.

5.7.1.2.1.4 Adverse events causing processing suspension

A list of adverse events that caused a permanent treatment to be discontinued, sorted by subject, date of occurrence, and time.

5.7.1.2.2 Allergic reactions 5.7.1.2.2.1 List of allergic reactions

Written any detailed incidents related to allergic reactions with adverse supplemental information as adverse events.

Regardless of the definition of the treatment period, a list of individual data is provided, sorted by subject, date of occurrence and time (as distinguished from all adverse event lists, see paragraph 5.7.1.2.1.1), including descriptions of specific adverse events, Symptoms, possible etiology, measures taken, measures of vital signs (when they occur, during and after recovery), and descriptions of allergic or allergic reactions.

The assessment of all such cases is also presented in accordance with the Allergic Response Assessment Committee (ARAC), including, in particular, whether the reported event constitutes an allergic reaction and, if so, its diagnosis and severity.

All cases are described in detail in the clinical study report.

5.7.1.2.2.2 Subject and familial allergy medical history

Subjects and familial allergic medical history were documented in writing for any subject with a potential allergic reaction, coded according to MedDRA Version 13.0 and listed by subject.

5.7.1.2.3 Suspected pancreatitis

A written case of any suspected pancreatitis was recorded in detail as an adverse event.

Regardless of the definition of the processing period, a list of individual data is provided, sorted by subject, date of occurrence, and time (different from all adverse events) For a list, see paragraph 5.7.1.2.1.1), including descriptions of specific adverse events, values of amylase and lipase, assessment by gastroenterologists, and possible causes of pancreatitis.

All cases are described in detail in the clinical study report.

5.7.1.2.4 Clinical laboratory evaluation 5.7.1.2.4.1 Hematology and biochemical data

Clinical laboratory safety (hematology, biochemistry, and urine testing) was assessed on Day 1 of Treatment Sessions 1 and 2 and at the end of the study visit. Based on the study schedule, it is not planned to evaluate these safety parameters during the processing period.

Baseline values are the values collected on day 1 of each treatment period. If any subject repeats any of the planned baseline tests, the last re-checked value is considered baseline, provided that the test is performed prior to the first study of the product and under the same conditions (eg, fasting glucose) get on.

The raw data for amylase and lipase were summarized in narrative statistics according to treatment group and time point.

Provide the following list:

‧ Individual data lists for subjects with post-baseline clinically significant abnormalities (PCSA), categorized by function and measurement time;

• A list of all individual data for hematology and biochemistry, including re-checking values and data for non-project laboratory tests, based on biological function and measurement time. In these lists, individual data is indicated if it is below or above the laboratory lower or upper limit and/or when the absolute boundary of the PCSA standard is reached (if any);

‧ Liver function data of subjects who have experienced at least one of the following conditions List of: - At least one of ALT > 3x ULN and total bilirubin > 2x ULN after initial administration during the study, regardless of the definition of the treatment period; - identical conjugated bilirubin after the first administration >35% total bilirubin and total bilirubin >1.5x ULN, regardless of the definition of the treatment period.

‧If available, increase with ALT 2x ULN-related list, including details of medication information, medical and surgical history, alcohol habits, predisposing factors, related signs, and symptoms;

‧ A list that is outside the definition of the normal range.

5.7.1.2.4.2 Urine test data

List all qualitative and quantitative urine test results (measuring rods), including values for re-checking.

5.7.1.2.5 Signs of life 5.7.1.2.5.1 Heart rate and blood pressure

At the first day of treatment periods 1 and 2 and the end of study visit, heart rate and systolic and diastolic blood pressure (SBP and DBP) were measured after 10 minutes in the resting position and 3 minutes after the standing position. Based on the study schedule, it is not planned to evaluate these safety parameters during the processing period.

The values to be used as baselines are the D1 evaluation values for each treatment period. If any of the subjects repeats any of the planned baseline tests, the final re-checked values are considered baselines, provided that the tests were performed prior to the first study product submission.

Regarding heart rate and blood pressure, raw data (upright position and standing posture) are summarized in narrative statistics according to measurement type (posture), parameters, and time points.

The following list is provided: • All individual data, including non-planned and re-checked values, are listed by measurement type (upright, standing, upright). In the list, if the absolute boundary of the PCSA standard is reached (if defined), individual values are indicated; ‧ separate lists of individual data for subjects with post-baseline PCSA.

5.7.1.2.5.2 Body temperature

List all individual data.

5.7.1.2.6 ECG

The ECG is automatically read on the first day of each treatment period and at the end of the study visit. According to the research schedule, there is no plan to evaluate the treatment period.

The heart rate and PR, QRS, QT and corrected QT interval (QTc) from the 12-lead ECG automatic reading were analyzed as raw parameter values; the baseline values were the values on the first day of each period. If any of the subjects repeat any of the planned baseline trials, the final rechecked values are considered baseline, provided that the tests are performed prior to the first drug administration.

For all parameters, the raw data was summarized in narrative statistics by treatment group and time point.

The following list is available: • All individual data, including re-checked values, according to the list of measurement types. In the list, if the boundary of the PCSA standard is reached (bounded Marked values; ‧ separate lists of individual data for subjects with post-baseline PCSA; ‧ use all post-dose time points with extended QTc (male > 450 ms, female > 470 ms) or A separate list of QTc's cardiac markers (if any) relative to baseline changes >60 ms (if any); ‧ at least one abnormality (ie abnormality) in quality assessment after initial dosing List of subjects for ECG).

5.7.1.3 Analysis of pharmacokinetic data

All pharmacokinetic analyses were performed using the pharmacokinetic population.

Day of dosing by the lixisenatide plasma concentration data measured at least the following pharmacokinetic parameters using non-compartmental _{methods: C max, t max, AUC} last, AUC and t _{1 / 2z.}

According to narrative statistics (number of observations (N), arithmetic mean and geometric mean, standard deviation (SD), standard deviation of mean (SEM), coefficient of variation (CV%), median, minimum and maximum The value, as well as the number of observations, summarizes the pharmacokinetic parameters.

To support PK/PD analysis, the early partial AUC (AUC _t1-t2 ) was calculated, where t1t2 was 02h, 1-2h and 1h30.

5.7.1.4 Pharmacokinetic/pharmacodynamic analysis

Not applicable.

5.7.2 Other statistical/analytical considerations

Not applicable.

5.7.3 Deciding the sample size

Using a 2x2 crossover design, a total of 20 completed subjects (each The order of 10) requires GBEF at 60 minutes to confirm that lixisenatide is no less than 20% absolute difference (no inferiority limit) for placebo, with a test power of 90%, if the actual SD in the subject is 0.10% And assume that the actual difference between placebo and lixisenatide is up to 0.09%. To allow withdrawal, 24 subjects participated in the study.

6 study subjects 6.1 Subject arrangement

A total of 24 subjects were enrolled, randomized, and exposed to study treatment (Table 5), and all subjects completed 2 study periods.

6.2 Program deviation (PROTOCOL DEVIATIONS) 6.2.1 Deviations associated with selected criteria and cause exclusion from pharmacodynamic analysis

There were no deviations from the protocol that led to exclusion from pharmacodynamic analysis. A deviation associated with non-compliance with inclusion criteria I05 (QRS = 122 msec) is not considered clinically relevant.

6.2.2 Random grouping and dosing irregularities

There were no random group irregularities during the study. All 24 subjects received the study product (lixilax, placebo) and were enrolled in the radiolabel and CCK-8.

According to the pharmacokinetic pattern of subject No. 826001012, it seems Not exposed to lixisenatide during any test period. Conduct some research to eliminate technical issues. These studies are described in the file notes.

‧ Exclude another subject receiving lixisenat to replace subject No. 826001012; no other subjects in this dosing group had unusual lixisenatide exposure.

‧ The site confirms in writing that the drug delivery system is being carried out correctly. Check all medications and complete the medication instructions before use.

‧ Re-analyze the drug 0000644 of batch IP0000988 and confirm the product characteristics of each drug.

‧Because no reasonable explanation was found for this finding, subjects were included in pharmacodynamics and pharmacokinetic analysis.

6.2.3 Other deviations

No other important deviations were observed.

6.3 Blindness

No bluntness during the study.

6.4 Analyzed data sets

The safety, pharmacodynamics, and pharmacokinetic groups included 24 subjects (Table 6).

6.5 Demographics and other baseline characteristics 6.5.1 Demography

Demographic data on safety populations are summarized in Table 7. There were 15 males and 9 female subjects aged 35 to 62 years (mean age ± SD: 47.8 ± 7.9 years) and an average body mass index of approximately 26 kg/m ² . All subjects were Caucasian except that one subject was black.

The demographic characteristics of male and female subjects are summarized in Table 8.

6.5.2 Medical history

The medical history obtained during screening is for inclusion only; no relevant medical history has been recorded.

6.5.3 Disease characteristics at baseline

Not applicable. The study recruited healthy subjects.

6.5.4 Other baseline characteristics

no

6.5.5 Prior and / or combined drug treatment

The previous medication was not stopped before the study began. No combined drug treatment was administered during the study.

6.6 Processing compliance measurements

During the two trial periods, 24 subjects were scheduled to receive radiolabeling, CCK8 infusion, and lixisenatide or placebo treatment for 1 day. For all subjects, the duration of the CCK-8 infusion corresponds to the 60 minute duration specified by the protocol.

7 Pharmacodynamic evaluation 7.1 Pharmacodynamic results 7.1.1 Main pharmacodynamic variables (variable) - Gallbladder excretion rate at 60 minutes

For the primary assessment (GBEF at 60 minutes), the mean difference between the placebo and lixisenatide was 45.80% (95% CI: 29.92; 61.68). The upper limit of the confidence interval is more than 20%, indicating that Lisila has not been confirmed to be inferior to placebo (Table 9).

According to the pharmacokinetic profile of a subject (No. 826001012), it does not appear to be exposed to lixisenatide during any of the trial periods. There may be no reasonable explanation, so the subject's data has been included in the pharmacodynamics community. However, for information purposes, the exclusion of the subject from the analysis had the least impact on the results.

GBEF evaluation time corresponds to the time after the start of CCK8 infusion

LSM = least square difference, CI = confidence interval

Number of subjects: N=24 received placebo, N=24 received lixisenatide 20 μg

^a average value provided by LSM

^b mean difference = LSM (placebo) - LSM (Lisila to 20 μg)

7.1.1.1 Secondary analysis 7.1.2 Secondary pharmacodynamic variables 7.1.2.1 Gallbladder discharge rate at 30 minutes

For the secondary assessment index (GBEF at 30 minutes), the parameterized mean difference between placebo and lixisenatide was 41.43% (95% CI: 28.64; 54.23) (Table 10).

Table 17 shows GBEF at 30 minutes with no data for subjects who did not appear to be exposed to lixisenatide during any of the trial periods (see paragraph 7.1.1).

GBEF evaluation time corresponds to the time after the start of CCK8 infusion

LSM = least square difference, CI = confidence interval

Number of subjects: N=24 received placebo, N=24 received lixisenatide 20 μg

^a average value provided by LSM

^b mean difference = LSM (placebo) - LSM (Lisila to 20 μg)

7.1.2.2 Narrative statistics at 30 minutes and 60 minutes

The narrative statistics for GBEF are summarized in Table 11.

The mean (SEM) GBEF (%) at 30 minutes and 60 minutes after placebo administration were 59.80 (5.67) and 84.95 (4.20), respectively. The average GBEF was 17.97 (3.35) and 39.01 (5.85) at 30 minutes and 60 minutes after a single dose of lixisenatide (Table 11).

The GBEF box whiskers at 30 minutes and 60 minutes after placebo and lixisenatide are presented in Figure 7.

GBEF = gallbladder discharge rate

GBEF evaluation time corresponds to the time after the start of CCK-8 infusion

7.1.2.3 Narrative statistics in 2 minutes

A graph of the mean of every two minutes of each treatment and the median GBEF during the CCK-8 infusion is provided in Figure 4. Leesila's GBEF (%) was lower than placebo at all time points and was more than placebo Slowly increase. Both curves show a gradual increase over time.

7.1.3 pharmacodynamic conclusion

Compared to placebo, a single administration of 20 μg of lixisenatide significantly reduced the GB emptying (expressed as GBEF (%)) due to CCK8 response by 45.8% at 45 minutes (absolute difference: 95% CI: 29.92; 61.68). It has not been confirmed that lixisela is not inferior to placebo.

The design of this study and the sample size calculations are based on the methodology reported in the open literature (5). Using the same CCK8 infusion dose and duration, this published study reported that in normal healthy individuals, the mean percentage of GBEF at 30 minutes and 60 minutes were 64% (±23%) and 84% (±16%), respectively. In the PDY11431 study, 23 of the 24 subjects receiving placebo had consistent GBEF values as expected and at normal concentrations (~40% at 60 minutes). The exception was Subject No. 826001001, which showed only 17% GBEF at 60 minutes after administration of placebo followed by CCK8 infusion. This does not seem to indicate a relevant functional disorder in this subject because the GBEF receiving lixisenatide was 75.1% at 60 minutes.

When receiving lixisenatide, 22 of 24 subjects had a lower GBEF at 60 minutes than a placebo at 60 minutes; for another, lisilla The GBEF is the same as the placebo. Of the 24 subjects, 13 had a GBEF below the lower limit of normal (~40%) at 60 minutes.

8 Safety assessment 8.1 Exposure

All 24 included subjects received a single period during each study period A second dose of 20 μg lixisenatide or placebo was completed as planned.

8.2 Adverse events 8.2.1 Brief summary of adverse events

TEAEs without severe TEAE, treatment of sudden SAE, or discontinuation of study treatment were reported during the study.

At the time of lixisenatide treatment, 4 of the 24 subjects (16.7%) experienced at least 1 TEAE compared to 1 of 24 subjects treated with placebo (4.2%). All TEAEs are mild or moderate.

There were no reports of allergic reactions requiring an allergic response assessment committee and no suspected pancreatitis.

8.2.2 Display of adverse events

The number and percentage of subjects with TEAE are summarized in Table 13 by treatment group, major system organ class, and preferred language. All adverse events are provided in accordance with the preferred language and the original language description used by the researcher.

Table 13 - Number of subjects (%) who deal with sudden adverse events by major systemic organ class and preferred language - Safety ethnic group

8.2.3 Analysis of adverse events

The TEAE of subjects receiving lixisenatide was mainly gastrointestinal disease SOC. The most frequently reported TEAE was nausea, which was reported in 3 subjects (Table 13). All TEAEs are mild or moderate.

One subject (No. 826001005) experienced a mild vomiting event that occurred approximately 5 hours after lixisenatide administration and 4 hours after the start of CCK infusion, and was assessed to be associated with lixisenatide and CCK8 treatment. Symptoms last about 5 minutes and are recovered without treatment.

One subject (No. 826001021) experienced a hematoma at the site of infection 1 day after the administration of Lisila, which was thought to be related to lixisenatide. The hematoma was mild and gradually recovered over the next 11 days.

One subject (No. 826001013) experienced abdominal pain 6 days after the administration of Lisila and was considered a post-treatment adverse event. Note that the subject had a deep breath at the right costal edge and a palpation at the time of the follow-up physical examination. Degree tenderness. Subjects were not found to have any abdominal discomfort except for deep palpation. The remaining abdominal examinations were evaluated as normal in the safety laboratory. After 2 days, the symptoms were completely restored without treatment. This incident was considered by the researchers to be related to Lisila and CCK-8.

Based on the assessment of skin reaction measures, no adverse events caused an overall score of ≧3.

A total of 8 subjects were written to record 10 adverse events during the study period: 1 pre-treatment adverse event, 6 treatment for sudden adverse events, and 3 post-treatment adverse events. The most common adverse events were gastrointestinal disorders (5 adverse events), followed by general illnesses (2 adverse events), injuries, poisoning and procedural disorders (2 adverse events), and skin and subcutaneous tissue disorders (1 adverse event) ).

A list of adverse events (preferred words) is provided below by subject (treatment group).

‧ Subject No. 826001002: Contusion (before treatment)

‧ Subject No. 826001005: nausea, vomiting (Lisila)

‧ Subject No. 826001007: Disgusting (Lisila)

‧ Subject No. 826001009: Swelling of the catheter site (placebo)

‧ Subject No. 826001013: Abdominal Pain (Lisila)

‧ Subject No. 826001015: Corneal abrasions (placebo)

‧ Subject No. 826001021: Hematoma at the injection site (Lisila)

‧ Subject No. 826001024: Blood blisters (placebo), nausea (Lisila)

8.3 Death, critical adverse events, and other significant adverse events 8.3.1 Death

No deaths were reported during the study period.

8.3.2 Critical adverse events

Critical adverse events were not reported during the study period.

8.3.3 Adverse events or other significant adverse events that caused withdrawal

Adverse events leading to withdrawal from the study, as well as significant adverse events, were not reported.

8.4 Clinical Laboratory Assessment

A list of subjects with all laboratory values and all possible clinically significant abnormalities (PCSA) laboratory values.

8.4.1 Laboratory values during the time

Analyze individual data for laboratory values only.

8.4.2 Individual subject changes in laboratory values

There were a few PCSAs in potassium, glucose, neutrophils, and eosinophils on day -1 of Day 2 and at the end of the study. Very few subjects have >2x ULN lipase values, but the re-checked values are within the normal range. PCSA reported without liver function, renal function, platelets or coagulation.

8.4.3 Individual clinically relevant abnormalities in laboratory data

Clinically relevant abnormalities of laboratory data were not reported during the study.

8.5 Vital signs, body surveys and other safety observations 8.5.1 Signs of life

A list of subjects providing all vital sign data.

8.5.1.1 Life signs in time

Provides narrative statistics of vital sign data.

8.5.1.2 Individual subject changes in vital signs

One subject randomized to placebo-Lisila (No. 826001004) experienced a decrease in the supineness of the SBP at the end of the study (standing-upper SBP ≦ 20 mm Hg). No other PCSA was observed during the study.

8.5.1.3 Individual clinically relevant abnormalities of vital signs

No clinically relevant abnormalities of vital signs were reported during the study.

8.5.2 ECG

A list of subjects providing all ECG data.

8.5.2.1 ECG values during the time

Provides narrative statistics of ECG data.

8.5.2.2 Changes in individual subjects of the ECG

None of the subjects had PCSA with QTc ≧ 500 ms (Table 14).

Subject No. 826001006, a 44-year-old male with a normal QTc of 418 ms at the baseline of Phase 2. Four (4) days later he received a single subcutaneous injection of 20 μg of lixisenatide, and he had a non-symptomatic PCSA with a prolonged QTc of 452 ms at the end of the study (re-check value 455 ms), corresponding to Increased by 34 ms relative to baseline (Table 14).

Subject No. 826001017, a 43-year-old male, received a placebo-lixilax, with a non-symptomatic PCSA with a prolonged QTc of 459 ms at baseline 2 (re-check value 436 ms). The QTc value at the end of the study was normal (409 ms).

8.5.2.3 Individual clinically relevant abnormalities of the electrocardiogram

No clinically relevant abnormalities in ECG values were reported during the study.

8.6 Security Conclusion

In summary, 20 μg of lixisenatide was administered tolerate well in healthy subjects. There were no critical adverse events and no study was withdrawn due to TEAE.

One of the 24 subjects (16.7%) experienced at least 1 TEAE when treated with lixisenatide compared to 1 of 24 subjects treated with placebo (4.2%). All TEAEs were mild and recovered without improved treatment. The TEAE of subjects administered to lixisenatide was primarily gastrointestinal SOC, such as nausea (3 out of 24 subjects) and vomiting (1 out of 24 subjects).

There were no clinically significant changes in laboratory parameters. No PCSA was observed in liver function and renal function.

There were no clinically significant findings in vital signs and ECG parameters. No subjects had QTc ≧ 500 ms. A male subject who received lixisenatide during the second period had an extended QTc of 452 ms at the end of the study, corresponding to an increase of 34 ms relative to baseline (4 days after Lisila was administered). Another subject who received a placebo during Phase 1 had a QTc of +459 ms on Day -1 of Day 2. The QTc values at the end of the study were normal (409 ms).

Adverse events related to allergic reactions or suspected pancreatitis have not been reported.

9 Pharmacokinetic assessment 9.1 Plasma concentration

Bioanalytical reports on the analysis of Lisila can be obtained at a later date. Unlike the original plan, all samples from the placebo group were incorrectly analyzed.

Individual lixisenatide plasma concentrations and narrative statistics were provided. Individual plasmas of Lisirasa versus time were calculated and a plot of plasma concentration of lixisenatide versus time was calculated.

All blood samples were collected within 15% of the planned sampling time.

Except for one case, the sample taken prior to administration in Lisila had a lower concentration of lixisenatide (12 pg/mL) than LLOQ (measured before the dose of subject No. 82600101> LLOQ (19.9 pg) /mL)). Two patients receiving placebo showed a detached value > LLOQ (subject No. 826001005 at time 1 hour 30 minutes, 12.1 pg/mL and subject 826001023 at time 10 hours) , 18.2 pg/mL), while other measurable forms The sample is <LLOQ.

For subjects No. 826001012 during the two trial periods (placebo and Lisila), all samples were below LLOQ and no reason could be found. Therefore, this subject was unable to calculate the pharmacokinetic parameters.

The average (SD) plasma lixisenatide concentration-time profile processed by lixisenatide is shown in Figure 5 (linear scale) and Figure 6 (semi-log scale).

9.2 Pharmacokinetic parameters

Individual pharmacokinetic analysis of the plasma Pharmacokinetic data obtained after a single administration of 20 μg of lixisenatide and corresponding narrative statistics were provided.

A narrative statistical summary of the pharmacological data of lixisenatide is provided in Table 15.

After subcutaneous administration of 20 μg of lixisenatide, the mean peak exposure (C _max ) was 104 pg/mL and appeared after 2 hours (median). The total exposure (AUC) was 634 pg*h/mL on average, and the mean exposure during the interval of primary evaluation of gallbladder emptying (AUC _12h ) was 87.3 pg*h/mL.

9.3 Pharmacokinetic/pharmacodynamic relationship

Not applicable

9.4 Pharmacokinetic conclusion

After subcutaneous administration of 20 μg of lixisenatide, the mean peak exposure (C _max ) was 104 pg/mL and appeared after 2 hours (median). The total exposure (AUC) was 634 pg*h/mL on average, and the mean exposure during the interval of primary evaluation of gallbladder emptying (AUC _12h ) was 87.3 pg*h/mL.

10 Discussion and general conclusions

In this placebo-control crossover study, subcutaneous administration of a single dose of lixisenatide 20 μg was significantly reduced in response to CCK-8 at 30 minutes and 60 minutes in healthy subjects and was expressed as GBEF (%) ) GB emptying. Lisila did not confirm inferiority after 60 minutes of infusion of CCK-8 on placebo.

Treatment with lixisenatide produced maximum plasma concentrations 2 hours after injection. The total exposure (AUC) was 634 pg*h/mL on average, and the mean exposure during the interval of primary evaluation of gallbladder emptying (AUC _12h ) was 87.3 pg*h/mL.

Lisila was generally well tolerated and evaluated as safe in 24 subjects. The most frequent adverse events are nausea. There are no serious or critical adverse events.

11 References

1. Kakhi VR, Zakavi SY, Davoudi Y. Normal values of gallbladder ejection fraction using 99mTc-sestamibi scintigraphy after a fatty meal formula. J Gastrointestin Liver Dis. 2007 Jun; 16(2): 157-61. Pubmed PMID: 17592562.

2. Kao CH, Tsou CT, Wang SJ, Yeh SH. The evaluation of gallbladder function by quantitative radionuclide cholescintigraphy in patients with noninsulin-dependent diabetes mellitus. Nucl Med Commun. 1993 Oct; 14(10): 868-72. PubMed PMID : 8233230.

3. Prandini N. Methods of measuring gallbladder motor functions-the need for standardization: scintigraphy. Dig Liver Dis. 2003 Jul; 35 Suppl 3: S62-6. PubMed PMID: 12974513.

4. Krishnamuithy GT, Krishnamurthy S, Brown PH. Constancy and variability of gallbladder ejection fraction: impact on diagnosis and therapy. J Nucl Med. 2004 Nov; 45(11): 1872-7. PubMed PMID: 15534057.

5. Ziessman HA, Tulchinsky M, Lavely WC, Gaughan JP, Allen TW, Maru A, et al. Sincalide-stimulated cholescintigraphy: a multicenter investigation to determine optimal infusion methodology and gallbladder ejection fraction normal values. J Nucl Med. 2010 Feb; 51(2): 277-81. Epub 2010 Jan 15. PubMed PMID: 20080900.

6. Burdick R, Graybill F. (1992). Confidence intervals on variance components. Marcel Dekker, NY. ISBN 0-8247-8644-0..

12 Supporting information

Figure 1: Schematic description of the study design. Refer to the detailed study flow chart for physical examination, vital signs, laboratory tests, ECG, and AE assessment. Abbreviations: CCK-8 = cholecystokinin-8; IP = research product; EOS = end of study.

Figure 2: Examples of gallbladder filling curves and emptying curves for normal healthy subjects receiving placebo and lixisenatide. Horizontal coordinate: time (minutes). Vertical coordinates: count per second (CPS).

Figure 3: Example of GBEF (%) of normal healthy subjects receiving placebo and lixisenatide. Horizontal coordinate: time (minutes). Vertical coordinates: GBEF (%).

Figure 4: Mean and median individual maps - pharmacodynamics population.

Figure 5: Mean (+SD) lixisenatide plasma concentration (linear scale).

Figure 6: Mean plasma concentration of Lisila (half log scale).

Figure 7: Boxer's chart of GBEF at 30 minutes and 60 minutes (placebo on the right and Lisila to 20 μg on the left). The evaluation time for GBEF corresponds to the time after the start of the CCK-8 infusion. Symbol: box = Q1-Q3 quartile; + = average; box line: median; required length = maximum ≦ 1.5 (Q3-Q1); star = deviation of the subject identifier. Abbreviations: GBEF = gallbladder discharge rate; min = minutes.

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Claims (22)

A pharmaceutical composition for treating a disease or condition, wherein the disease or condition is associated with (a) stenosis or/and occlusion in the biliary tract, or/and (b) biliary dyskinesia, the composition comprising at least one GLP -1 agonist and pharmaceutically acceptable carrier, diluent or/and auxiliary substance as appropriate. For example, the pharmaceutical composition of claim 1 wherein the stenosis or/and obstruction is located in the hepatic duct, in the gallbladder, in the cystic duct, in the common bile duct, in the hepatopancreatic duct, in the nipple or in the edi In the sphincter. The pharmaceutical composition according to any one of claims 1 to 2, wherein the obstruction is caused by stones, stone disease or/and biliary mud in the biliary tract. For example, the pharmaceutical composition of claim 3, wherein the stone disease is biliary stone or/and cholelithiasis. The composition of any one of claims 1 to 2, wherein the disease or condition is cancer. The composition of claim 5, wherein the cancer is located in the hepatic duct, in the gallbladder, in the cystic duct, in the common bile duct, in the hepatic pancreatic duct, in the nipple or in the eddy sphincter. The composition of claim 5 or 6, wherein the cancer comprises a tumor. A composition according to any one of the preceding claims, wherein the stenosis or/and occlusion causes pain. A composition according to any one of the preceding claims, wherein the treatment is a palliative treatment. The composition of any of the preceding claims, wherein the treatment is treatment for at least one month, at least two months, at least three months, at least four months, or at least six months. A composition according to any one of the preceding claims, wherein the GLP-1 agonist is selected from the group consisting of GLP-1, analogs and derivatives thereof, exenatide-3, analogs and derivatives thereof, A group consisting of cenapeptide-4, its analogs and derivatives. The composition of any one of claims 1 to 11, wherein the GLP-1 agonist is lixisenatide or/and a pharmaceutically acceptable salt thereof. The composition of any one of claims 1 to 11, wherein the GLP-1 agonist is liraglutide or/and a pharmaceutically acceptable salt thereof. A pharmaceutical composition for palliative treatment comprising at least one GLP-1 agonist and, optionally, a pharmaceutically acceptable carrier, diluent or/and auxiliary. The pharmaceutical composition of claim 14, wherein the palliative treatment comprises treating pain caused by (a) stenosis or/and occlusion in the biliary tract, or/and (b) biliary dyskinesia. A pharmaceutical composition for treating pain, the composition comprising at least one GLP-1 agonist and optionally a pharmaceutically acceptable carrier Agent, diluent or / and auxiliary substances. The pharmaceutical composition of claim 16 wherein the pain is associated with (a) stenosis or/and obstruction in the biliary tract, or/and (b) biliary dyskinesia. The pharmaceutical composition of claim 16 or 17, wherein the pain treatment is palliative treatment. A method of treating a disease or condition associated with (a) stenosis or/and occlusion in the biliary tract, or/and (b) biliary dyskinesia, the method comprising comprising at least one GLP-1 agonist, and optionally The pharmaceutical composition selected as a pharmaceutically acceptable carrier, diluent or/and auxiliary substance is administered to an individual in need thereof. The method of claim 19, wherein the disease or condition is defined in any one of claims 2 to 10. A GLP-1 agonist for the manufacture of a medicament for treating a disease or condition associated with (a) stenosis or/or occlusion in the biliary tract, or/and (b) biliary dyskinesia, the medicament comprising at least one GLP-1 agonist, and optionally a pharmaceutically acceptable carrier, diluent or/and auxiliary substance. The use of claim 20, wherein the disease or condition is defined in any one of claims 2 to 10.