WO2021136293A1 - 胰岛素衍生物 - Google Patents

胰岛素衍生物 Download PDF

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Publication number
WO2021136293A1
WO2021136293A1 PCT/CN2020/141018 CN2020141018W WO2021136293A1 WO 2021136293 A1 WO2021136293 A1 WO 2021136293A1 CN 2020141018 W CN2020141018 W CN 2020141018W WO 2021136293 A1 WO2021136293 A1 WO 2021136293A1
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Prior art keywords
γglu
human insulin
desb30 human
insulin
diacyl
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PCT/CN2020/141018
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English (en)
French (fr)
Chinese (zh)
Inventor
甘忠如
陈伟
张一宁
薛方凯
蔡玲玉
牛江红
穆彬
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Gan and Lee Pharmaceuticals Co Ltd
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Gan and Lee Pharmaceuticals Co Ltd
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Priority to KR1020227024018A priority Critical patent/KR20220121833A/ko
Priority to US17/758,101 priority patent/US20240239862A1/en
Priority to EP20910566.7A priority patent/EP4086279A4/en
Priority to CN202410312576.5A priority patent/CN118666989A/zh
Priority to CN202410312489.XA priority patent/CN118480114A/zh
Priority to CA3166495A priority patent/CA3166495A1/en
Priority to MX2022008174A priority patent/MX2022008174A/es
Priority to AU2020418205A priority patent/AU2020418205A1/en
Priority to CN202080091196.3A priority patent/CN114901682B/zh
Application filed by Gan and Lee Pharmaceuticals Co Ltd filed Critical Gan and Lee Pharmaceuticals Co Ltd
Priority to CN202410254905.5A priority patent/CN118420744A/zh
Priority to JP2022540767A priority patent/JP2023510206A/ja
Priority to BR112022013150A priority patent/BR112022013150A2/pt
Publication of WO2021136293A1 publication Critical patent/WO2021136293A1/zh
Anticipated expiration legal-status Critical
Priority to US19/052,262 priority patent/US20250228919A1/en
Priority to JP2025128114A priority patent/JP2025161829A/ja
Ceased legal-status Critical Current

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/22—Hormones
    • A61K38/28—Insulins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/02—Inorganic compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
    • A61K47/10—Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/575—Hormones
    • C07K14/62—Insulins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • the present invention relates to the field of therapeutic peptides, in particular to novel insulin derivatives, their pharmaceutical preparations, their pharmaceutical compositions with long-acting GLP-1 compounds, their pharmaceutical compositions with fast-acting insulin, and the insulin derivatives , Medicinal use of pharmaceutical preparations and pharmaceutical compositions.
  • Insulin is a polypeptide hormone secreted by the beta cells of the pancreas.
  • Insulin is composed of two polypeptide chains named A chain and B chain, which are connected by two interchain disulfide bonds.
  • a chain and B chain In human, porcine and bovine insulin, the A chain and B chain contain 21 and 30 amino acid residues, respectively. However, between different species, there are differences in the amino acid residues present in different positions of the two chains.
  • the widespread use of genetic engineering has made it possible to prepare analogs of naturally occurring insulin by replacing, deleting and adding one or more amino acid residues.
  • Insulin can be used to treat diabetes and the diseases associated with or caused by it, and is necessary to maintain normal metabolic regulation.
  • natural insulins such as human insulin have a short action time, which necessitates frequent injections of patients, causing many injection-related discomforts. Therefore, people have been striving to obtain insulin derivatives or analogs with good efficacy, longer action time, and lower injection frequency to improve the inconvenience and discomfort caused by higher frequency insulin injections.
  • WO1995007931A1 discloses the marketed long-acting insulin detemir. Its molecular structure is characterized by the removal of the threonine at position 30 of the B chain of human insulin, and a 14-carbon fat monomer attached to the lysine residue at position 29 of the B chain. acid.
  • WO2005012347A2 discloses another long-acting insulin degludec, which has been on the market. Insulin degludec is a new type of ultralong-acting insulin that has a longer action time than insulin detemir. Its molecular structure is characterized by the removal of the 30th position of the human insulin B chain.
  • Amino acid the 16-carbon fatty diacid side chain is connected to the lysine residue at position B29 through a glutamic acid molecule.
  • CN101573133B and WO2009/010428 disclose PEG (PEGylated) extended insulin, which has a longer action time than conventional unmodified insulin.
  • WO2013086927A1 and WO2018/024186 disclose a long-acting acylated derivative of a human insulin analog.
  • the present invention provides novel insulin derivatives (such as acylated insulin).
  • novel insulin derivatives such as acylated insulin.
  • the new insulin derivatives for example, acylated insulin
  • the marketed insulin degludec trade name "Novota”
  • certain other insulin derivatives have Unexpected significantly increased efficacy or efficacy, longer duration of action, longer half-life in vivo, good bioavailability, better safety, and more satisfactory physical and chemical stability Sex, and solubility.
  • the present invention provides an insulin derivative comprising an insulin parent, an albumin binding residue, and a linking group Lin, the insulin parent being a naturally-occurring insulin or an insulin analog, the insulin The protein binding residue is connected to the insulin parent via the linking group Lin, wherein,
  • the linking group Lin has at least 10, preferably at least 15, preferably at least 25, preferably at least 30, preferably at least 36, preferably 15-100, preferably 25-90, preferably 30-80, Preferably, a hydrophilic linking group of 30-59, preferably 30-54 carbon atoms; or, the linking group Lin contains at least 5 neutral amino acid residues containing alkylene glycol; The linking group Lin contains at least 6 neutral amino acid residues containing alkylene glycol; preferably, the linking group Lin contains 5-9 neutral amino acid residues containing alkylene glycol.
  • the linking group Lin comprises an alkylene glycol having at least 15, preferably at least 20, preferably at least 24, preferably 15-50, preferably 20-39 carbon atoms, and
  • the albumin binding residue contains 20-40 carbon atoms, preferably the albumin binding residue is a linear or branched lipophilic group containing 20-40 carbon atoms, preferably the albumin binding residue Is a fatty acid or aliphatic diacid containing 20-26 carbon atoms (more preferably a fatty acid or aliphatic diacid of 20-24 carbon atoms), in which the hydroxyl group has changed formally from the carboxyl group of the fatty acid and the aliphatic diacid One of the carboxyl groups of the diacid is removed.
  • the inventors unexpectedly discovered that the combination of a certain length of albumin binding residues and a certain length of hydrophilic linking groups in the insulin derivatives of the present invention makes the insulin derivatives of the present invention relative to the existing ones.
  • Insulin derivatives while having an equivalent or longer action time, also have unexpectedly significantly increased pharmacodynamics, and when albumin is present, the ability to bind insulin receptors is significantly reduced by the influence of albumin. It has significantly improved binding capacity to insulin receptor.
  • the insulin parent comprises at least one lysine residue
  • the albumin binding residue is connected to the amino group of the lysine residue or the N-terminal amino acid residue of the insulin parent via the linking group Lin. Phase connection.
  • the insulin derivative further comprises one or more linking group II, the linking group II is an acidic amino acid residue, and the linking group II is connected to the albumin binding residue And the linking group Lin, and/or between the linking group Lin and the insulin parent; preferably, the linking group II is connected between the albumin binding residue and the link Between the group Lin.
  • an insulin derivative which is an acylated insulin
  • the insulin parent of the acylated insulin is a naturally-occurring insulin or an insulin analogue, and contains at least one lysine residue
  • the acyl moiety of the acylated insulin is connected to the lysine residue of the insulin parent or the amino group of the N-terminal amino acid residue, and the acyl moiety is represented by formula (A):
  • n 0, 6, 7, 8 or 9;
  • I is a neutral amino acid residue containing alkylene glycol
  • III is a fatty acid or aliphatic diacid containing 20-26 (preferably 20-24) carbon atoms, wherein the hydroxyl group has been formally removed from one of the carboxyl group of the fatty acid and the carboxyl group of the aliphatic diacid;
  • the acyl moiety is represented by formula (A'):
  • n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and n'is an integer
  • I' is a neutral amino acid residue containing alkylene glycol
  • III is a fatty acid or aliphatic diacid containing 20-26 (preferably 20-24) carbon atoms, wherein the hydroxyl group has been formally removed from one of the carboxyl group of the fatty acid and the carboxyl group of the aliphatic diacid;
  • Number (I ') n' is from 15 to 100 total carbon atoms, preferably 20 to 100, preferably 25-90, preferably 30-80, preferably 30-59, preferably 30-54 months.
  • an insulin derivative which is an acylated insulin
  • the insulin parent of the acylated insulin is a naturally-occurring insulin or an insulin analogue, and contains at least one lysine residue
  • the acyl moiety of the acylated insulin is connected to the lysine residue of the insulin parent or the amino group of the N-terminal amino acid residue, and the acyl moiety is represented by formula (A):
  • n is 5, 6, 7, 8 or 9;
  • I is a neutral amino acid residue containing alkylene glycol
  • III is an aliphatic diacid containing 20-26 (preferably 20-24) carbon atoms, wherein the hydroxyl group has been formally removed from one of the carboxyl groups of the aliphatic diacid;
  • the acyl moiety is represented by formula (A'):
  • n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and n'is an integer
  • I' is a neutral amino acid residue containing alkylene glycol
  • III is an aliphatic diacid containing 20-26 (preferably 20-24) carbon atoms, wherein the hydroxyl group has been formally removed from one of the carboxyl groups of the aliphatic diacid;
  • Number (I ') n' is 25 to 90 total carbon atoms, preferably 30-80, preferably 30-59, preferably 30-54 months.
  • n is preferably, n is 5, 6, 7, or 8; and/or
  • n is an integer of 1-6, preferably m is 1, 2, 3, or 4, preferably m is 1 or 2, preferably m is 1;
  • III is an aliphatic diacid containing 20-26 (preferably 20-23) carbon atoms, preferably III is an aliphatic diacid containing 20, 21, or 22 carbon atoms, wherein the hydroxyl group has been formally removed from the One of the carboxyl groups of the aliphatic diacid; and/or
  • the insulin matrix contains a lysine residue.
  • I is: -HN-(CH 2 ) 2 -O-(CH 2 ) 2 -O-CH 2 -CO-, -HN-(CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -CO-, -HN-(CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -CO-, -HN-(CH 2 ) 2 -O-( CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -CO-, -HN-(CH 2 ) 2
  • II is an amino acid residue selected from: ⁇ Glu, ⁇ Glu, ⁇ Asp, ⁇ Asp, ⁇ -D-Glu, ⁇ -D-Glu, ⁇ -D-Asp or ⁇ -D-Asp, preferably, II is selected from ⁇ Glu Or ⁇ Asp; and/or
  • III is HOOC-(CH 2 ) 18 -CO-, HOOC-(CH 2 ) 19 -CO-, HOOC-(CH 2 ) 20 -CO-, HOOC-(CH 2 ) 21 -CO-, HOOC-(CH 2 ) 22 -CO-, or HOOC-(CH 2 ) 24 -CO-, preferably, III is HOOC-(CH 2 ) 18 -CO-, HOOC-(CH 2 ) 19 -CO-, HOOC-(CH 2 ) 20 -CO-, HOOC-(CH 2 ) 21 -CO- or HOOC-(CH 2 ) 22 -CO-, preferably III is HOOC-(CH 2 ) 18 -CO-, HOOC-(CH 2 ) 20 -CO- or HOOC-(CH 2 ) 22 -CO-.
  • the formula (A) is connected to the amino group of the lysine residue or the N-terminal amino acid residue of the insulin parent through the C-terminus of I or the formula (A') is connected to the C-terminus of I'through the The lysine residue of the insulin parent or the amino group of the N-terminal amino acid residue is connected.
  • the acyl moiety is linked to the epsilon amino group of the lysine residue of the insulin parent.
  • the lysine residue of the insulin parent is located at position B29.
  • the insulin parent is selected from the following insulins or insulin analogues: desB30 human insulin (SEQ ID NO: 1 and SEQ ID NO: 2, respectively representing the A chain and the B chain); A14E, B16H, B25H ,desB30 human insulin (SEQ ID NO: 3 and SEQ ID NO: 4, representing the A chain and B chain, respectively); A14E, B16E, B25H, desB30 human insulin (SEQ ID NO: 5 and SEQ ID NO: 6, respectively, representing A chain and B chain); human insulin (SEQ ID NO: 7 and SEQ ID NO: 8, respectively representing the A chain and B chain); A21G human insulin (SEQ ID NO: 9 and SEQ ID NO: 10, respectively representing A Chain and B chain); A21G, desB30 human insulin (SEQ ID NO: 11 and SEQ ID NO: 12, representing the A chain and B chain, respectively); or B28D human insulin (SEQ ID NO: 13 and SEQ ID NO: 14, Denote A chain and B chain respectively);
  • the insulin parent is selected from the following insulins
  • the acylated insulin is selected from the following insulins: B29K(N( ⁇ )-eicosandioyl- ⁇ Glu-5xOEG), desB30 human insulin; B29K(N( ⁇ )-eicosandi Acyl- ⁇ Glu-6xOEG), desB30 human insulin; B29K (N( ⁇ )-eicosanedioyl- ⁇ Glu- ⁇ Glu-5xOEG), desB30 human insulin; B29K(N( ⁇ )-eicosanedioyl- ⁇ Glu- ⁇ Glu-6xOEG), desB30 human insulin; B29K (N( ⁇ )-eicosane diacyl-5xOEG- ⁇ Glu), desB30 human insulin; B29K (N( ⁇ )-eicosane diacyl-6xOEG- ⁇ Glu), desB30 Human insulin; B29K (N( ⁇ )-eicosane diacyl-6xOEG- ⁇ Glu), desB30 Human insulin; B29K (N( ⁇ )-eicos
  • the acylated insulin is selected from the following insulins: B29K(N( ⁇ )-eicosandioyl- ⁇ Glu-5xOEG), desB30 human insulin; B29K(N( ⁇ )-eicosandi Acyl- ⁇ Glu-6xOEG), desB30 human insulin; B29K (N( ⁇ )-docosanedioyl- ⁇ Glu-5xOEG), desB30 human insulin; B29K(N( ⁇ )-docosanedioyl- ⁇ Glu- 6xOEG), desB30 human insulin; B29K(N( ⁇ )-eicosane diacyl- ⁇ Glu-7xOEG), desB30 human insulin; B29K(N( ⁇ )-eicosane diacyl- ⁇ Glu-8xOEG), desB30 human insulin ; B29K(N( ⁇ )-docosanedioyl- ⁇ Glu-7xOEG), desB30 human insulin; B29K(N( ⁇ )-docosaned
  • the acylated insulin is selected from the following insulins: B29K(N( ⁇ )-eicosandioyl- ⁇ Glu-5xOEG), desB30 human insulin; B29K(N( ⁇ )-eicosandi Acyl- ⁇ Glu-6xOEG), desB30 human insulin; B29K (N( ⁇ )-docosanedioyl- ⁇ Glu-5xOEG), desB30 human insulin; B29K(N( ⁇ )-docosanedioyl- ⁇ Glu- 6xOEG), desB30 human insulin; B29K (N( ⁇ )-eicosane diacyl- ⁇ Glu-8xOEG), desB30 human insulin; B29K (N( ⁇ )-docosane diacyl- ⁇ Glu-8xOEG), desB30 human Insulin; A14E, B16H, B25H, B29K (N( ⁇ )-eicosane diacyl- ⁇ Glu-6xOEG), desB30 human insulin; A14E, B
  • a pharmaceutical composition comprising the insulin derivative of the present invention as described above and one or more pharmaceutically acceptable excipients.
  • the pharmaceutical composition contains at least 1.5 moles of zinc ions/6 moles of insulin derivatives; preferably at least 2.2 moles of zinc ions/6 moles of insulin derivatives; preferably at least 3.5 moles of zinc ions/6 moles Insulin derivatives; preferably containing at least 4.5 moles of zinc ions/6 moles of insulin derivatives; preferably containing 4.5-12 moles of zinc ions/6 moles of insulin derivatives; more preferably containing 4.5-10 moles of zinc ions/6 moles of insulin Derivatives; more preferably containing 4.5-8 moles of zinc ions/6 moles of insulin derivatives; more preferably containing 4.5-7.5 moles of zinc ions/6 moles of insulin derivatives; more preferably containing 4.5-7.0 moles of zinc ions/6 moles Insulin derivatives; more preferably containing 4.5-6.5 moles of zinc ions/6 moles of insulin derivatives; and/or
  • the pharmaceutical composition has a pH of 6.5-8.5; preferably a pH of 6.8-8.2; preferably a pH of 7.0-8.2; preferably a pH of 7.2-7.6; more preferably a pH of 7.4 or 7.6.
  • the pharmaceutical composition further comprises glycerin, phenol, m-cresol, NaCl, and/or Na 2 HPO 4 ; preferably, the pharmaceutical composition further comprises glycerin, phenol, and NaCl; preferably, the The pharmaceutical composition further comprises glycerin, phenol, m-cresol, and NaCl; preferably, the pharmaceutical composition further comprises glycerin, phenol, NaCl and Na 2 HPO 4 ; more preferably, the pharmaceutical composition further comprises glycerin, phenol , M-cresol, NaCl and Na 2 HPO 4 .
  • the glycerin content does not exceed about 2.5% (weight/weight), preferably does not exceed about 2% (weight/weight), preferably about 0.3% to about 2% (weight/weight), preferably From about 0.5% to about 1.8% (weight/weight), preferably from about 0.7% to about 1.8% (weight/weight), more preferably from about 1% to about 1.8% (weight/weight); and/or
  • the content of the phenol is about 16 to 80 mM, preferably about 25-75 mM, preferably about 45-70 mM, preferably about 45-65 mM; preferably about 45 mM, about 46 mM, about 47 mM, about 48 mM, about 49 mM, about 50mM, about 51mM, about 52mM, about 53mM, about 54mM, about 55mM, about 56mM, about 57mM, about 58mM, about 59mM, about 60mM, about 61mM, about 62mM, about 63mM, about 64mM, or about 65mM; and/ or
  • the content of the m-cresol is about 0-35mM, preferably about 0-19mM, preferably about 0-15mM, preferably about 0mM, about 1mM, about 2mM, about 3mM, about 4mM, about 5mM, about 6mM, About 7mM, about 8mM, about 9mM, about 10mM, about 11mM, about 12mM, about 13mM, about 14mM, or about 15mM; and/or
  • the content of the NaCl is about 0-150 mM, preferably about 5-120 mM, preferably about 10-120 mM, preferably about 10-100 mM, more preferably about 10-75 mM, more preferably about 10-50 mM, more preferably Is about 10-30mM; and/or
  • the content of the Na 2 HPO 4 is about 0-75 mM, preferably about 5-60 mM, preferably less than about 50 mM, more preferably less than about 25 mM, more preferably less than about 15 mM; and/or
  • the content of the acylated insulin is higher than about 0.3 mM, preferably higher than about 0.6 mM, preferably about 0.3-12 mM, preferably about 0.6-9.0 mM, preferably about 0.6-8.4 mM, preferably about 0.6-7.2 mM, preferably about 0.6-6.0mM, preferably about 0.6-4.2mM, preferably about 0.6-3.6mM, preferably about 0.6-3.0mM, preferably about 0.6-2.4mM, preferably about 0.6-2.1mM, Preferably it is about 0.6-1.2 mM.
  • the insulin derivative is B29K(N( ⁇ )-eicosanedioyl- ⁇ Glu-5xOEG), desB30 human insulin; B29K(N( ⁇ )-eicosanedioyl- ⁇ Glu-6xOEG ), desB30 human insulin; B29K(N( ⁇ )-docosanedioyl- ⁇ Glu-5xOEG), desB30 human insulin; B29K(N( ⁇ )-docosanedioyl- ⁇ Glu-6xOEG), desB30 human Insulin; B29K (N( ⁇ )-eicosandioyl- ⁇ Glu-8xOEG), desB30 human insulin; B29K (N( ⁇ )-docosanedioyl- ⁇ Glu-8xOEG), desB30 human insulin; A14E, B16H , B25H, B29K (N( ⁇ )-eicosanedioyl- ⁇ Glu-6xOEG), desB30 human insulin; or A14E, B
  • a pharmaceutical composition which comprises about 0.6-4.2 mM of the insulin derivative of the present invention as described above, about 1% to about 1.8% (weight/weight) of glycerol, about 45- 65 mM phenol, about 4.5-6.5 moles of zinc ions/6 moles of insulin derivatives, about 10-120 mM sodium chloride, about 0-15 mM m-cresol, and have a pH of about 7.0-8.2, preferably, the Insulin derivatives are B29K (N( ⁇ )-eicosanedioyl- ⁇ Glu-5xOEG), desB30 human insulin; B29K (N( ⁇ )-eicosanedioyl- ⁇ Glu-6xOEG), desB30 human insulin; B29K( N( ⁇ )-docosanedioyl- ⁇ Glu-5xOEG), desB30 human insulin; B29K(N( ⁇ )-docosanedioyl- ⁇ Glu-6xOEG),
  • a pharmaceutical composition comprising about 0.6 mM or 1.2 mM of the insulin derivative of the present invention as described above, 1.7% (weight/weight) of glycerol, about 45 mM of phenol, about 10 mM Of m-cresol, about 6.5 moles of zinc ion/6 moles of insulin derivative, about 20 mM of sodium chloride, and a pH of about 7.0-8.0.
  • the insulin derivative is B29K(N( ⁇ )- Eicosane diacyl- ⁇ Glu-5xOEG), desB30 human insulin; B29K (N( ⁇ )-eicosane diacyl- ⁇ Glu-6xOEG), desB30 human insulin; B29K(N( ⁇ )-docosane diacyl - ⁇ Glu-5xOEG), desB30 human insulin; B29K(N( ⁇ )-docosanedioyl- ⁇ Glu-6xOEG), desB30 human insulin; B29K(N( ⁇ )-eicosanedioyl- ⁇ Glu-8xOEG) ,desB30 human insulin; B29K(N( ⁇ )-docosane diacyl- ⁇ Glu-8xOEG), desB30 human insulin; A14E, B16H, B25H, B29K(N( ⁇ )-eicosane diacyl- ⁇ Glu-6xOEG ), desB30 human insulin; B29K(N( ⁇ )-e
  • the insulin derivative is B29K(N( ⁇ )-eicosanedioyl- ⁇ Glu-6xOEG), desB30 human insulin; B29K(N( ⁇ )-docosanedioyl- ⁇ Glu-6xOEG), desB30 human insulin; A14E, B16H, B25H, B29K (N( ⁇ )-eicosanedioyl- ⁇ Glu-6xOEG), desB30 human insulin; B29K(N( ⁇ )-eicosanedioyl- ⁇ Glu-12xPEG), desB30 human insulin or A14E, B16H, B25H, B29K (N( ⁇ )-docosanedioyl- ⁇ Glu-6xOEG), desB30 human insulin.
  • a pharmaceutical composition comprising about 0.6-4.2mM of the insulin of the present invention as described above, about 1% to about 2% (preferably about 1.5%-1.7%) (weight/weight) ) Glycerol, about 15mM-60mM (preferably about 30mM-60mM, more preferably about 45mM-60mM) phenol, about 1.5-7.0 (preferably about 2.2-4.5) moles of zinc ions/6 moles of insulin derivatives, about 10- 120mM (preferably about 20-50mM) sodium chloride, about 0-25mM (preferably about 0-10mM) m-cresol, and having a pH of about 7.0-8.2, preferably, the insulin derivative is B29K(N( ⁇ )-Eicosane diacyl- ⁇ Glu-5xOEG), desB30 human insulin; B29K(N( ⁇ )-Eicosane diacyl- ⁇ Glu-6xOEG), desB30 human insulin; B29K(N( ⁇ )-22 Alkanedioyl
  • a pharmaceutical composition comprising about 0.6-4.2mM of the insulin of the present invention as described above, about 1.5%-1.7% (weight/weight) of glycerol, and about 45-60mM of phenol , About 2.2-4.5 moles of zinc ions/6 moles of insulin derivatives, about 20mM sodium chloride, about 0-10mM m-cresol, and have a pH of about 7.0-8.0, preferably, the insulin derivative is B29K (N( ⁇ )-eicosane diacyl- ⁇ Glu-5xOEG), desB30 human insulin; B29K(N( ⁇ )-eicosane diacyl- ⁇ Glu-6xOEG), desB30 human insulin; B29K(N( ⁇ )- Dodecanedioyl- ⁇ Glu-5xOEG), desB30 human insulin; B29K(N( ⁇ )-docosanedioyl- ⁇ Glu-6xOEG), desB30 human insulin; B29K(N( ⁇ )-docosanedi
  • the pharmaceutical composition further comprises an insulinotropic GLP-1 compound; preferably, the pharmaceutical composition further comprises an insulinotropic GLP-1 compound selected from: N- ⁇ 26 -(17- Carboxyhexadecanoylamino)-4(S)-carboxybutyryl-[Arg34]GLP-1-(7-37) peptide, N- ⁇ 26 -(17-carboxyheptadecanylamino)-4(S )-Carboxybutyryl-[Gly8,Arg34]GLP-1-(7-37) peptide, N- ⁇ 26 -[2-(2-[2-(2-[2-[2-[2-[2-[2-[4-(17 -Carboxyheptadecanylamino)-4(S)-carboxybutyrylamino]ethoxy)ethoxy)acetamino)ethoxy]ethoxy)acetyl][Aib8,Arg34]GLP-1- (7-37) peptide, N- ⁇ 26 -[2-
  • the pharmaceutical composition further comprises the insulinotropic GLP-1 compound represented by formula (B), or a pharmaceutically acceptable salt, amide or ester thereof:
  • G1 is a GLP-1 analogue with Arg at position 34 corresponding to GLP-1(7-37) (SEQ ID NO: 15) and Ala or Gly at position 8
  • [Acy-(L1) r -(L2) q ] is a substituent attached to the epsilon amino group of the Lys residue at position 26 of the GLP-1 analog, wherein
  • r is an integer of 1-10
  • q is an integer of 0 or 1-10
  • Acy is an aliphatic diacid containing 20-24 carbon atoms, in which the hydroxyl group has been formally removed from one of the carboxyl groups of the aliphatic diacid;
  • L1 is an amino acid residue selected from: ⁇ Glu, ⁇ Glu, ⁇ Asp, ⁇ Asp, ⁇ -D-Glu, ⁇ -D-Glu, ⁇ -D-Asp or ⁇ -D-Asp;
  • L2 is a neutral amino acid residue containing alkylene glycol
  • G1 is [Gly8, Arg34] GLP-1-(7-37) peptide (SEQ ID NO: 16) or [Arg34] GLP-1-(7-37) peptide (SEQ ID NO: 17), preferably [Gly8 ,Arg34]GLP-1-(7-37) peptide; and/or
  • r is 1, 2, 3, 4, 5 or 6, preferably r is 1, 2, 3 or 4, preferably r is 1 or 2, preferably r is 1;
  • q is 0, 1, 2, 3, 4, 5, 6, 7, or 8, preferably, q is 0, 1, 2, 3, or 4, more preferably, q is 0, 1, or 2;
  • Acy is an aliphatic diacid containing 20-23 carbon atoms, preferably Acy is an aliphatic diacid containing 20, 21, or 22 carbon atoms, in which the hydroxyl group has changed formally from among the carboxyl groups of the aliphatic diacid One is removed.
  • L2 is: -HN-(CH 2 ) 2 -O-(CH 2 ) 2 -O-CH 2 -CO-, -HN-(CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -CO-, -HN-(CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -CO-, -HN-(CH 2 ) 2 -O-( CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -CO-, -HN-(CH 2 )
  • L1 is selected from ⁇ Glu or ⁇ Asp, preferably L1 is ⁇ Glu; and/or
  • Acy is HOOC-(CH 2 ) 18 -CO-, HOOC-(CH 2 ) 19 -CO-, HOOC-(CH 2 ) 20 -CO-, HOOC-(CH 2 ) 21 -CO- or HOOC-(CH 2 ) 22 -CO-, preferably, Acy is HOOC-(CH 2 ) 18 -CO-, HOOC-(CH 2 ) 20 -CO- or HOOC-(CH 2 ) 22 -CO-.
  • Acy, L1, and L2 in formula (B) are sequentially connected by an amide bond, and the C-terminus of L2 is connected to the epsilon amino group of the Lys residue at position 26 of the GLP-1 analog.
  • the insulinotropic GLP-1 compound is selected from the following compounds:
  • the insulinotropic GLP-1 compound is selected from the following compounds:
  • the insulinotropic GLP-1 compound is:
  • the pharmaceutical composition or combined preparation of the insulin derivative (e.g., acylated insulin) and insulinotropic GLP-1 compound of the present invention not only does not weaken the insulin derivative (e.g., acylated insulin). ), and the combination preparation has better physical stability than the single-drug preparation.
  • the physical stability of the combined preparation of the present invention is unexpected.
  • the combined preparation also increases the chemical stability of the insulin derivative (for example, acylated insulin) compared to the single-drug preparation.
  • the pharmaceutical composition further comprises fast-acting insulin.
  • the fast-acting insulin is selected from one or more of Asp B28 human insulin, Lys B28 Pro B29 human insulin, Lys B3 Glu B29 human insulin, human insulin and desB30 human insulin; preferably, The fast-acting insulin is Asp B28 human insulin, Lys B28 Pro B29 human insulin, Lys B3 Glu B29 human insulin, human insulin or desB30 human insulin.
  • the molar ratio of the insulin derivative to the fast-acting insulin is about 60:3 to about 0.5:3, preferably about 57:3 to about 1:3, preferably about 55: 3 to about 1.2:3, preferably about 50:3 to about 1.5:3, preferably about 40:3 to about 1.5:3, preferably about 30:3 to about 1.5:3, preferably about 27:3 to About 1.5:3, preferably about 25:3 to about 1.5:3, preferably about 22:3 to about 1.5:3, preferably about 20:3 to about 1.5:3, preferably about 17:3 to about 1.5 :3, preferably about 15:3 to about 1.5:3, preferably about 12:3 to about 1.5:3, preferably about 10:3 to about 1.5:3, preferably about 9:3 to about 1.5:3 , Preferably from about 8:3 to about 1.5:3, preferably from about 7:3 to about 1.5:3, preferably from about 6.9:3 to about 1.5:3, preferably from about 6.8:3 to about 1.5:3, preferably From about 6.5:3 to about 1.5:3,
  • the inventors unexpectedly found that the pharmaceutical composition comprising the insulin derivative (for example, acylated insulin) of the present invention and the double insulin component of insulin aspart is compared with the double insulin comprising deglubber and insulin aspart after administration.
  • the pharmaceutical composition of the insulin component has an unexpectedly increased hypoglycemic effect.
  • the dosage ratio of the insulin derivative (such as acylated insulin) and insulin aspart of the present invention is much smaller than that of insulin deglubber and insulin aspart. When the dosage is compared, it can still achieve a better or equivalent hypoglycemic effect.
  • the insulin derivative is the insulin derivative of the present invention as described above; preferably, the insulin derivative is B29K (N( ⁇ )-eicosanedioyl- ⁇ Glu-5xOEG), desB30 Insulin; B29K (N( ⁇ )-eicosandioyl- ⁇ Glu-6xOEG), desB30 human insulin; B29K (N( ⁇ )-docosanedioyl- ⁇ Glu-5xOEG), desB30 human insulin; B29K(N ( ⁇ )-docosane diacyl- ⁇ Glu-6xOEG), desB30 human insulin; B29K(N( ⁇ )-eicosane diacyl- ⁇ Glu-7xOEG), desB30 human insulin; B29K(N( ⁇ )-two Decanediacyl- ⁇ Glu-8xOEG), desB30 human insulin; B29K(N( ⁇ )-docosanediacyl- ⁇ Glu-7xOEG), desB30 human insulin; B29K(N( ⁇ )-two Decan
  • the pharmaceutical composition comprises about 0.09-0.36 mM insulin derivative, about 0.18 mM Asp B28 human insulin, about 0.85% to about 2.0% (weight/weight) glycerol, about 15-70 mM Phenol, about 8-14 moles of zinc ions/6 moles of insulin derivatives, about 10-120 mM sodium chloride, about 0-15 mM m-cresol, and have a pH of about 7.0-8.2, wherein the insulin derivative
  • the substance is B29K (N( ⁇ )-eicosanedioyl- ⁇ Glu-5xOEG), desB30 human insulin; B29K (N( ⁇ )-eicosanedioyl- ⁇ Glu-6xOEG), desB30 human insulin; B29K(N( ⁇ )-docosane diacyl- ⁇ Glu-5xOEG), desB30 human insulin; B29K (N( ⁇ )-docosane diacyl- ⁇ Glu-6xOEG), desB30 human insulin; B29K (
  • the pharmaceutical composition comprises about 0.165-0.18mM insulin derivative, about 0.18mM Asp B28 human insulin, about 1.5%-1.7% (weight/weight) glycerol, about 20mM-30mM Phenol, about 9-12 moles of zinc ion/6 moles of insulin derivative, about 20mM-75mM sodium chloride, about 10mM-15mM m-cresol, and having a pH of about 7.0-8.2, wherein the insulin derivative B29K(N( ⁇ )-eicosanedioyl- ⁇ Glu-5xOEG), desB30 human insulin; B29K(N( ⁇ )-eicosanedioyl- ⁇ Glu-6xOEG), desB30 human insulin; B29K(N( ⁇ ) )-Docosane diacyl- ⁇ Glu-5xOEG), desB30 human insulin; B29K(N( ⁇ )-docosane diacyl- ⁇ Glu-6xOEG), desB30 human insulin; B29K(N( ⁇ )-
  • the insulin derivative or the pharmaceutical composition described above of the present invention is used as a medicine.
  • the insulin derivative or the pharmaceutical composition described above of the present invention is used as a medicament for treating or preventing diabetes, hyperglycemia, and/or impaired glucose tolerance.
  • the insulin derivative or the pharmaceutical composition described above of the present invention is used to treat or prevent diabetes, hyperglycemia, and/or impaired glucose tolerance.
  • the use of the aforementioned insulin derivative or the pharmaceutical composition of the present invention in the preparation of a medicine is provided; preferably, the medicine is used for the treatment or prevention of diabetes, hyperglycemia, and/or glucose Impaired tolerance.
  • the medicament is used to treat diabetes, and the insulin derivative is administered to the same patient every other day or less frequently, and on average, for a period of at least 1 month, 6 months, or 1 year , The insulin is not administered to the same patient more frequently.
  • the drug is used to treat diabetes, and the insulin derivative is administered twice a week or less frequently, and on average, during a period of at least 1 month, 6 months, or 1 year, The insulin derivatives are not administered to the same patient more frequently.
  • the drug is used to treat diabetes, and the insulin derivative is administered once a week or less frequently, and on average, during a period of at least 1 month, 6 months, or 1 year, The insulin derivatives are not administered to the same patient more frequently.
  • the insulin derivatives (for example, acylated insulin) of the present invention have long pharmacokinetics (hereinafter also referred to as PK) characteristics, making it possible to subcutaneously treat diabetic patients twice a week, once a week or less frequently.
  • PK pharmacokinetics
  • the present invention provides a method for treating or preventing diabetes, hyperglycemia, and/or impaired glucose tolerance, comprising administering a therapeutically effective amount of the insulin derivative of the present invention as described above or the pharmaceutical composition.
  • the present invention provides a method for improving insulin receptor binding ability of insulin derivatives in the presence of albumin, which includes:
  • the insulin derivative is obtained by linking the albumin binding residue with the naturally-occurring insulin or insulin analog via the linking group Lin, wherein the linking group Lin has at least 10, preferably at least 15, preferably at least 25 One, preferably at least 30, preferably at least 36, preferably 15-100, preferably 25-90, preferably 30-80, preferably 30-59, preferably 30-54 hydrophilic linking groups of carbon atoms
  • the albumin binding residue contains 20-40 carbon atoms, preferably the albumin binding residue contains a linear or branched lipophilic group with 20-40 carbon atoms, preferably the albumin binding residue It is a fatty acid or aliphatic diacid containing 20-26 carbon atoms (more preferably a fatty acid or aliphatic diacid of 20-24 carbon atoms), in which the hydroxyl group has changed formally from the carboxyl group of the fatty acid or the aliphatic diacid Removed from one of the carboxyl groups of the diacid, or
  • the derivative is obtained by modifying naturally-occurring insulin or insulin analogues with formula (A) or formula (A'),
  • n 0, 6, 7, 8 or 9;
  • I is a neutral amino acid residue containing alkylene glycol
  • III is an albumin binding residue containing a linear or branched lipophilic group with 20-40 carbon atoms, preferably III is a fatty acid or aliphatic diacid containing 20-26 carbon atoms (more preferably 20-24 Carbon-atom fatty acid or aliphatic diacid), wherein the hydroxyl group has been formally removed from one of the carboxyl group of the fatty acid and the carboxyl group of the aliphatic diacid;
  • (A') is III-(II) m -(I') n' -(A'),
  • n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and n'is an integer
  • I' is a neutral amino acid residue containing alkylene glycol
  • III is an albumin binding residue containing a linear or branched lipophilic group with 20-40 carbon atoms, preferably III is a fatty acid or aliphatic diacid containing 20-26 carbon atoms (more preferably 20-24 Carbon-atom fatty acid or aliphatic diacid), wherein the hydroxyl group has been formally removed from one of the carboxyl group of the fatty acid and the carboxyl group of the aliphatic diacid;
  • Number (I ') n' is from 15 to 100 total carbon atoms, preferably 20 to 100, preferably 25-90, preferably 30-80, preferably 30-59, preferably 30-54 months.
  • the present invention provides a method for improving the efficacy of insulin derivatives, which comprises: linking albumin binding residues with naturally-occurring insulin or insulin analogues via a linking group Lin to obtain the insulin derivatives.
  • the linking group Lin has at least 10, preferably at least 15, preferably at least 25, preferably at least 30, preferably at least 36, preferably 15-100, preferably 25-90, preferably 30- 80, preferably 30-59, preferably 30-54 hydrophilic linking groups;
  • the albumin binding residue contains 20-40 carbon atoms, preferably the albumin binding residue contains 20 A linear or branched lipophilic group with -40 carbon atoms, preferably the albumin binding residue is a fatty acid or aliphatic diacid containing 20-26 carbon atoms (more preferably a fatty acid with 20-24 carbon atoms or Aliphatic diacid), in which the hydroxyl group has been formally removed from one of the carboxyl group of the fatty acid or the carboxyl group of the aliphatic diacid, or
  • the derivative is obtained by modifying naturally-occurring insulin or insulin analogues with formula (A) or formula (A'),
  • n 0, 6, 7, 8 or 9;
  • I is a neutral amino acid residue containing alkylene glycol
  • III is an albumin binding residue containing a linear or branched lipophilic group with 20-40 carbon atoms, preferably III is a fatty acid or aliphatic diacid containing 20-26 carbon atoms (more preferably 20-24 Carbon-atom fatty acid or aliphatic diacid), wherein the hydroxyl group has been formally removed from one of the carboxyl group of the fatty acid and the carboxyl group of the aliphatic diacid;
  • (A') is III-(II) m -(I') n' -(A'),
  • n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, and n'is an integer
  • I' is a neutral amino acid residue containing alkylene glycol
  • III is an albumin binding residue containing a linear or branched lipophilic group with 20-40 carbon atoms, preferably III is a fatty acid or aliphatic diacid containing 20-26 carbon atoms (more preferably 20-24 Carbon-atom fatty acid or aliphatic diacid), wherein the hydroxyl group has been formally removed from one of the carboxyl group of the fatty acid and the carboxyl group of the aliphatic diacid;
  • Number (I ') n' is from 15 to 100 total carbon atoms, preferably 20 to 100, preferably 25-90, preferably 30-80, preferably 30-59, preferably 30-54 months.
  • n 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;
  • III is an aliphatic diacid containing 20-24 carbon atoms, in which the hydroxyl group has been formally removed from one of the carboxyl groups of the aliphatic diacid.
  • the naturally-occurring insulin or insulin analogue comprises at least one lysine residue, the linking group Lin, formula (A) or formula (A') and the lysine residue of the insulin parent The amino group or the amino group of the N-terminal amino acid residue is connected.
  • n is 5, 6, 7, or 8;
  • n 1, 2, 3, 4, 5 or 6, preferably m is 1, 2, 3, or 4, preferably m is 1 or 2, preferably m is 1;
  • III is an aliphatic diacid containing 20-26 (preferably 20-23) carbon atoms, preferably III is an aliphatic diacid containing 20, 21, or 22 carbon atoms, wherein the hydroxyl group has been formally removed from the One of the carboxyl groups of the aliphatic diacid; and/or
  • the insulin matrix contains a lysine residue.
  • I is: -HN-(CH 2 ) 2 -O-(CH 2 ) 2 -O-CH 2 -CO-, -HN-(CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -CO-, -HN-(CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -CO-, -HN-(CH 2 ) 2 -O-( CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -O-(CH 2 ) 2 -CO-, -HN-(CH 2 ) 2
  • II is an amino acid residue selected from: ⁇ Glu, ⁇ Glu, ⁇ Asp, ⁇ Asp, ⁇ -D-Glu, ⁇ -D-Glu, ⁇ -D-Asp or ⁇ -D-Asp, preferably, II is selected from ⁇ Glu Or ⁇ Asp; and/or
  • III is HOOC-(CH 2 ) 18 -CO-, HOOC-(CH 2 ) 19 -CO-, HOOC-(CH 2 ) 20 -CO-, HOOC-(CH 2 ) 21 -CO-, HOOC-(CH 2 ) 22 -CO-, or HOOC-(CH 2 ) 24 -CO-, preferably III is HOOC-(CH 2 ) 18 -CO-, HOOC-(CH 2 ) 20 -CO- or HOOC-(CH 2 ) 22 -CO-.
  • the formula (A) is connected to the amino group of the lysine residue or the N-terminal amino acid residue of the natural insulin or insulin analog through the C-terminus of I, or the formula (A') is connected through The C-terminus of I'is connected to the amino group of the lysine residue or the N-terminal amino acid residue of the natural insulin or insulin analog.
  • formula (A) or formula (A') is linked to the epsilon amino group of the lysine residue of the insulin parent.
  • the lysine residue of the natural insulin or insulin analog is located at position B29.
  • the naturally occurring insulin or insulin analogue is selected from the following insulin or insulin analogues: desB30 human insulin; A14E, B16H, B25H, desB30 human insulin; A14E, B16E, B25H, desB30 human insulin; Human insulin; A21G human insulin; A21G, desB30 human insulin; or B28D human insulin; preferably, the insulin parent is desB30 human insulin or A14E, B16H, B25H, desB30 human insulin.
  • Figure 1a shows the hypoglycemic effect of the compounds of Examples 1 and 2 of the present invention, insulin degludec, and vehicle on db/db mice.
  • Fig. 1b corresponding to Fig. 1a shows the AUC of the hypoglycemic effect of the compounds of Examples 1 and 2 of the present invention, insulin degludec and vehicle on db/db mice.
  • Figure 2a shows the hypoglycemic effects of the compounds of Examples 1 and 2 of the present invention, the compound of Comparative Example 2 and the vehicle on db/db mice.
  • Figure 2b corresponds to Figure 2a showing the AUC of the hypoglycemic effect of the compounds of Examples 1 and 2 of the present invention, the compound of Comparative Example 2 and the vehicle on db/db mice.
  • Figure 3a shows the hypoglycemic effect and action time of the compounds of Examples 1-3 of the present invention and the vehicle on db/db mice.
  • Figure 3b corresponds to Figure 3a showing the AUC of the hypoglycemic effect of the compounds of Examples 1-3 and the vehicle of the present invention on db/db mice.
  • Figure 4a shows the hypoglycemic effect and action time of the compound of Example 2 of the present invention, the compound of Comparative Example 3 and the vehicle on db/db mice.
  • Figure 4b corresponds to Figure 4a showing the AUC of the hypoglycemic effect of the compound of Example 2 of the present invention, the compound of Comparative Example 3 and the vehicle on db/db mice.
  • Figure 5a shows the hypoglycemic effect and action time of the compound of Comparative Example 3-4 of the present invention and the vehicle on db/db mice.
  • Figure 5b corresponds to Figure 5a showing the AUC of the hypoglycemic effect of the compound of Comparative Example 3-4 of the present invention and the vehicle on db/db mice.
  • Fig. 6a shows the hypoglycemic effect and action time of the compounds of Example 2 and Examples 4-5 of the present invention and the vehicle on db/db mice.
  • Figure 6b corresponds to Figure 6a showing the AUC of the hypoglycemic effect of the compounds of Examples 2 and 4-5 of the present invention and the vehicle on db/db mice.
  • Figure 7a shows the hypoglycemic effect of the compound of Example 1 of the present invention and the vehicle on streptozotocin (STZ)-induced type I diabetes (T1DM) rats.
  • Figure 7b corresponds to Figure 7a showing the AUC of the hypoglycemic effect of the compound of Example 1 of the present invention and the vehicle on STZ-induced type I diabetes (T1DM) rats.
  • Figure 8a shows the hypoglycemic effects of the title compounds of Comparative Example 5, Examples 15 and 16 of the present invention, and the vehicle on STZ-induced type I diabetes (T1DM) rats.
  • Figure 8b corresponds to Figure 8a showing the AUC of the hypoglycemic effect of Comparative Example 5, the title compounds of Examples 15 and 16, and the vehicle on STZ-induced type I diabetes (T1DM) rats.
  • Figure 9a shows the hypoglycemic effect of the compounds of Examples 2 and 4 of the present invention and the vehicle on STZ-induced type I diabetes (T1DM) female rats.
  • Figure 9b corresponds to Figure 9a showing the AUC of the hypoglycemic effect of the compounds of Examples 2 and 4 of the present invention and the vehicle on STZ-induced type I diabetes (T1DM) female rats.
  • Figure 10a shows the hypoglycemic effect of the title compounds of Comparative Example 5, Examples 15 and 16 and the vehicle of the present invention on db/db mice.
  • Figure 10b and Figure 10a correspondingly show the AUC of the hypoglycemic effect of Comparative Example 5, Examples 15 and 16 of the present invention, and the vehicle on the hypoglycemic effect of db/db mice.
  • Figure 11a shows the hypoglycemic effect of the title compound of Comparative Example 5 and Example 16 of the present invention and the vehicle on STZ-induced type I diabetes (T1DM) rats.
  • Fig. 11b and Fig. 11a correspondingly show the AUC of the hypoglycemic effect of the title compound of Comparative Example 5 and Example 16 of the present invention and the vehicle on STZ-induced type I diabetes (T1DM) rats.
  • Figure 12a shows insulin aspart, a pharmaceutical composition comprising insulin deglubber and insulin aspart dual insulin components, a pharmaceutical composition comprising the acylated insulin and insulin aspart dual insulin components of the present invention, and a vehicle pair STZ
  • T1DM induced type I diabetes
  • Figure 12b and Figure 12a correspondingly show insulin aspart, a pharmaceutical composition comprising insulin deglubber and insulin aspart dual insulin components, and a pharmaceutical composition comprising the acylated insulin and insulin aspart dual insulin components of the present invention
  • Figure 13a shows insulin aspart, a pharmaceutical composition comprising insulin deglubber and insulin aspart di-insulin components, a pharmaceutical composition comprising the acylated insulin and insulin aspart di-insulin components of the present invention, and a vehicle pair STZ
  • T1DM induced type I diabetes
  • Figure 13b and Figure 13a correspondingly show insulin aspart, a pharmaceutical composition containing deglubber and insulin aspart dual insulin components, and a pharmaceutical composition containing the acylated insulin and insulin aspart dual insulin components of the present invention
  • Figure 14a shows insulin aspart, a pharmaceutical composition comprising insulin deglubber and insulin aspart dual insulin components, a pharmaceutical composition comprising the acylated insulin and insulin aspart dual insulin components of the present invention, and the vehicle pair STZ Induced type I diabetes (T1DM) C57/6J mice before the fourth dose of mouse blood glucose.
  • T1DM vehicle pair STZ Induced type I diabetes
  • Figure 14b shows insulin aspart, a pharmaceutical composition comprising insulin deglubber and insulin aspart dual insulin components, a pharmaceutical composition comprising the acylated insulin and insulin aspart dual insulin components of the present invention, and the vehicle pair STZ Induced type I diabetes (T1DM) C57/6J mice before the eighth dose of mouse blood glucose.
  • T1DM vehicle pair STZ Induced type I diabetes
  • Figure 14c shows insulin aspart, a pharmaceutical composition comprising insulin deglubber and insulin aspart dual insulin components, a pharmaceutical composition comprising the acylated insulin and insulin aspart dual insulin components of the present invention, and a vehicle pair STZ Induced type I diabetes (T1DM) C57/6J mice before the tenth dose of mouse blood glucose.
  • T1DM STZ Induced type I diabetes
  • Figure 15a shows insulin aspart, a pharmaceutical composition comprising deglubber and insulin aspart dual insulin components, a pharmaceutical composition comprising the acylated insulin and insulin aspart dual insulin components of the present invention, and the vehicle pair STZ Induced type I diabetes (T1DM) C57/6J mice 1 hour after the fourth dose of mouse blood glucose.
  • T1DM vehicle pair STZ Induced type I diabetes
  • Figure 15b shows insulin aspart, a pharmaceutical composition comprising insulin deglubber and insulin aspart dual insulin components, a pharmaceutical composition comprising the acylated insulin and insulin aspart dual insulin components of the present invention, and the vehicle pair STZ Induced type I diabetes (T1DM) C57/6J mice with blood glucose 1 hour after the eighth dose.
  • T1DM vehicle pair STZ Induced type I diabetes
  • Figure 15c shows insulin aspart, a pharmaceutical composition comprising insulin deglubber and insulin aspart dual insulin components, a pharmaceutical composition comprising the acylated insulin and insulin aspart dual insulin components of the present invention, and a vehicle pair STZ Induced type I diabetes (T1DM) C57/6J mice 1 hour after the tenth dose of mouse blood glucose.
  • T1DM vehicle pair STZ Induced type I diabetes
  • Figure 16 shows insulin aspart, a pharmaceutical composition comprising insulin deglubber and insulin aspart dual insulin components, a pharmaceutical composition comprising the acylated insulin and insulin aspart dual insulin components of the present invention, and a vehicle pair STZ HbA1c-lowering effect in induced type I diabetes (T1DM) C57/6J mice.
  • Figure 17a shows the hypoglycemic effect of the compound of Example 4 of the present invention, insulin degludec and vehicle on STZ-induced type I diabetes (T1DM) rats.
  • Figure 17b corresponds to Figure 17a showing the AUC of the hypoglycemic effect of the compound of Example 4 of the present invention, insulin degludec and vehicle on STZ-induced type I diabetes (T1DM) rats.
  • Figure 18a shows a pharmaceutical composition comprising insulin deglubber and insulin aspart di-insulin components, a pharmaceutical composition comprising the acylated insulin and insulin aspart di-insulin components of the present invention, and a vehicle for type I induction of STZ
  • T1DM diabetic
  • Figure 18b and Figure 18a correspondingly show a pharmaceutical composition comprising insulin deglubber and insulin aspart dual insulin components, a pharmaceutical composition comprising the acylated insulin and insulin aspart dual insulin components of the present invention, and a vehicle pair AUC of the hypoglycemic effect of STZ-induced type I diabetes (T1DM) C57/6J mice.
  • T1DM STZ-induced type I diabetes
  • Figure 19 shows a pharmaceutical composition comprising insulin deglubber and insulin aspart dual insulin components, a pharmaceutical composition comprising the acylated insulin and insulin aspart dual insulin components of the present invention, and a vehicle for type I induction of STZ The effect of reducing HbA1c in diabetic (T1DM) C57/6J mice.
  • Figure 20a shows a pharmaceutical composition comprising insulin deglubber and insulin aspart dual insulin components, a pharmaceutical composition comprising the acylated insulin and insulin aspart dual insulin components of the present invention, and a vehicle for db/db mice The hypoglycemic effect.
  • Figure 20b and Figure 20a correspondingly show a pharmaceutical composition comprising insulin deglubber and insulin aspart double insulin components, a pharmaceutical composition comprising the acylated insulin and insulin aspart double insulin components of the present invention, and a vehicle pair AUC of hypoglycemic effect in db/db mice.
  • Figure 21a shows a pharmaceutical composition comprising insulin deglubber and insulin aspart di-insulin components, a pharmaceutical composition comprising the acylated insulin and insulin aspart di-insulin components of the present invention, and db/db small after the vehicle Random blood glucose of the rat.
  • Figures 21b and 21a correspondingly show after injection of a pharmaceutical composition containing deglubber and insulin aspart dual insulin components, a pharmaceutical composition containing the acylated insulin and insulin aspart dual insulin components of the present invention, and a vehicle AUC of random blood glucose in db/db mice.
  • Figure 21c shows a pharmaceutical composition comprising insulin deglubber and insulin aspart di-insulin components, a pharmaceutical composition comprising the acylated insulin and insulin aspart di-insulin components of the present invention, and db/db small after vehicle Fasting blood glucose of rats.
  • Figures 21d and 21c respectively show after injection of a pharmaceutical composition containing deglubber and insulin aspart dual insulin components, a pharmaceutical composition containing the acylated insulin and insulin aspart dual insulin components of the present invention, and a vehicle AUC of fasting blood glucose in db/db mice.
  • Figure 22 shows the receptor binding ability of the compound of Example 2 and the control compound 2 of the present invention in the presence of 2% HSA and 0% HSA, respectively.
  • insulin includes naturally-occurring insulin, such as human insulin, as well as its insulin analogs and insulin derivatives.
  • insulin analogue encompasses polypeptides that have one or more amino acid residues and/or addition of one or more amino acid residues that can be formally deleted and/or replaced (substitution) in natural insulin, and A molecular structure derived from the structure of a naturally occurring insulin, such as human insulin.
  • the added and/or substituted amino acid residues may be codable amino acid residues, or other naturally occurring amino acid residues, or purely synthetic amino acid residues.
  • the added and/or substituted amino acid residues are codable amino acid residues.
  • insulin derivative refers to a naturally-occurring insulin or insulin analogue that has been chemically modified.
  • the modification can be, for example, the introduction of a side chain or oxidation or reduction at one or more positions of the insulin backbone.
  • the groups of amino acid residues on insulin either convert free carboxyl groups into ester groups or acylate free amino groups or hydroxyl groups.
  • the acylated insulin of the present invention belongs to insulin derivatives.
  • insulin parent refers to an insulin derivative or the insulin part of an acylated insulin (also referred to herein as the parent insulin), for example, in the present invention refers to an insulin derivative or an acylated insulin that has no side chain attached or attached The acyl part.
  • the parent insulin may be a naturally occurring insulin, such as human insulin or porcine insulin.
  • the parent insulin may be an insulin analogue.
  • amino acid residue includes an amino acid from which a hydrogen atom has been removed from an amino group and/or a hydroxyl group has been removed from a carboxyl group and/or a hydrogen atom has been removed from a sulfhydryl group.
  • amino acid residues can be called amino acids.
  • amino acids mentioned herein are L-amino acids.
  • albumin binding residue refers to a residue capable of non-covalently binding to human serum albumin.
  • Albumin binding residues linked to insulin generally have a binding affinity for human serum albumin that is less than, for example, about 10 ⁇ M or even less than about 1 ⁇ M.
  • the albumin binding properties can be measured by the surface plasmon resonance described in the following literature: J. Biol. Chem. 277 (38), 35035-35042, (2002).
  • hydrophilic linking group refers to a linking group that contains at least 6 chemical moieties other than hydrogen atoms (moiety) that separates the insulin precursor from the albumin binding residues, and 30-50 of these non-hydrogen atoms % Is N or O.
  • Lipophilic refers to the ability of the group to dissolve in fats, oils, lipids, and lipophilic non-polar solvents such as hexane or toluene.
  • Lipophilic groups including but not limited to, for example, fats, fatty acids, fatty diacids, etc., usually have "lipid tails".
  • the lipid tails present in these lipophilic groups can be saturated or unsaturated, depending on Whether the lipid tail contains double bonds.
  • Lipid tails can also contain different lengths, such as tails having between 7-12 carbons (e.g., C 7-12 alkyl or C 7-12 alkenyl), tails having 13-22 carbons (e.g., C 13-12 alkyl or C 13-12 alkenyl), or a tail having 23-30 carbons (for example, C 23-30 alkyl or C 23-30 alkenyl).
  • tails having between 7-12 carbons e.g., C 7-12 alkyl or C 7-12 alkenyl
  • tails having 13-22 carbons e.g., C 13-12 alkyl or C 13-12 alkenyl
  • a tail having 23-30 carbons for example, C 23-30 alkyl or C 23-30 alkenyl.
  • alkylene glycol encompasses oligo- and polyalkylene glycol moieties as well as monoalkylene glycol moieties.
  • Monoalkylene glycols and polyalkylene glycols include, for example, chains based on mono-polyethylene glycol, mono-polypropylene glycol, and mono-polybutylene glycol, that is, based on the repeating unit -CH 2 CH 2 O-, -CH 2 CH 2 CH 2 O- or -CH 2 CH 2 CH 2 CH 2 O- chain.
  • the alkylene glycol moiety can be monodisperse (having a well-defined length/molecular weight) as well as polydisperse (having a less well-defined length/average molecular weight).
  • the monoalkylene glycol moiety includes -OCH 2 CH 2 O-, -OCH 2 CH 2 CH 2 O-, or -OCH 2 CH 2 CH 2 CH 2 O- containing different groups at each end.
  • fatty acid includes straight or branched chain aliphatic carboxylic acids, which have at least two carbon atoms and are saturated or unsaturated.
  • Non-limiting examples of fatty acids are, for example, myristic acid, palmitic acid, stearic acid, and arachidic acid.
  • aliphatic diacid includes linear or branched aliphatic dicarboxylic acids, which have at least two carbon atoms and are saturated or unsaturated.
  • Non-limiting examples of aliphatic diacids are adipic acid, suberic acid, sebacic acid, dodecanedioic acid, tetradecanedioic acid, hexadecanedioic acid, heptadecanedioic acid, octadecanedioic acid , Eicosandioic acid, Behenedioic acid, and tetracosanedioic acid.
  • fast-acting insulin includes fast-acting naturally-occurring insulin, insulin analogs, and insulin derivatives.
  • Fast-acting insulin usually starts to work in, for example, 1 to 20 minutes, reaches a peak in about an hour, and continues to work for three to five hours.
  • basic insulin means insulin that has a longer duration of action than normal or normal human insulin.
  • chemical stability means that, chemically, the insulin derivative of the present invention is sufficiently stable in the desired formulation. That is, only chemical degradation products are formed in an amount that does not impair the shelf life of the final drug product.
  • Chemical degradation products include deamidation products, isoaspartate formation, dimer formation, racemization products, products resulting from dehydration processes and the like. Chemical stability can be determined by HPLC analysis of aged samples or formulations.
  • binding ability to insulin receptor refers to the interaction between insulin and insulin receptor, and the magnitude or intensity of this interaction can be measured by, for example, surface plasmon resonance (SPR).
  • SPR surface plasmon resonance
  • when measuring with SPR when a solution containing insulin flows through a chip coated with insulin receptors, the interaction between insulin and insulin receptors will cause changes in the deflection angle of the SPR. This change is usually used as a relative response. Generally, the larger the relative response value, the higher the binding ability to insulin receptor.
  • High physical stability means that the tendency of fibrillation is less than 50% of that of human insulin. Fibrillation can be described by the lag time before fibril formation begins under given conditions.
  • Polypeptides with affinity for insulin receptor and IGF-1 receptor are polypeptides that can interact with insulin receptor and human IGF-1 receptor in a suitable binding assay. Such receptor assays are well known in the art.
  • pharmaceutical effect refers to the ability of a drug or active compound to produce a certain action or effect (for example, lowering blood sugar).
  • a drug or active compound for example, lowering blood sugar.
  • administering the same dose of the insulin derivative of the present invention will have a higher blood sugar lowering effect or effect than deglubber or other existing insulin derivatives.
  • diabetes includes type 1 diabetes, type 2 diabetes, gestational diabetes (during pregnancy) and other conditions that cause hyperglycemia.
  • the term is used for metabolic disorders, where the pancreas produces insufficient amounts of insulin, or where the cells of the body cannot respond appropriately to insulin, thereby preventing the cells from absorbing glucose. As a result, glucose accumulates in the blood.
  • Type 1 diabetes also known as insulin-dependent diabetes mellitus (IDDM) and juvenile diabetes
  • IDDM insulin-dependent diabetes mellitus
  • Type 2 diabetes also known as non-insulin-dependent diabetes mellitus (NIDDM) and adult-type diabetes, is associated with major insulin resistance and therefore relative insulin deficiency and/or major insulin secretion defects with insulin resistance.
  • GLP-1 analog refers to a peptide or compound that is a variant of human glucagon-like peptide-1 (GLP-1(7-37)), wherein One or more amino acid residues of GLP-1(7-37) are replaced, and/or one or more amino acid residues are deleted, and/or one or more amino acid residues are added.
  • GLP-1 (7-37) is shown in SEQ ID NO: 15 in the sequence list.
  • the peptide having the sequence shown in SEQ ID NO: 15 can also be referred to as "natural" GLP-1 or "natural” GLP-1 (7-37).
  • the amino acid residue numbering or position numbering of the GLP-1(7-37) sequence referred to herein is the sequence of His starting at position 7 and Gly ending at position 37.
  • GLP-1-(7-37) peptide which is a GLP with Gly and Arg at positions 8 and 34 corresponding to GLP-1(7-37) (SEQ ID NO: 15) -1 analogue.
  • [Arg34] GLP-1-(7-37) peptide is a GLP-1 analogue with Arg at the position corresponding to position 34 of GLP-1(7-37) (SEQ ID NO: 15).
  • the amino acid sequences of [Gly8,Arg34]GLP-1-(7-37) peptide and [Arg34]GLP-1-(7-37) peptide are as shown in SEQ ID NO: 16 and SEQ ID NO in the sequence list, respectively. : Shown at 17.
  • the term "derivative" as used herein refers to a chemically modified GLP-1 peptide or analogue in which one or more substituents have been covalently attached to the peptide .
  • Substituents can also be referred to as side chains.
  • the naming of insulin or GLP-1 compounds is based on the following principles: according to mutations and modifications (such as acylation) relative to human insulin, or mutations and modifications (such as acylation) of natural GLP-1 (7-37) ) Give a name.
  • mutations and modifications such as acylation
  • mutations and modifications such as acylation of natural GLP-1 (7-37)
  • the acyl moiety can be named according to the IUPAC nomenclature (OpenEye, IUPAC format). According to this nomenclature, the above-mentioned acyl moiety of the present invention is called the following name: [2-(2-[2-(2-[2-(2-[4-(19-carboxynonadenoylamino)-4 (S)-Carboxybutyrylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl], or [2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[
  • the insulin of Comparative Example 2 of the present invention is called "B29K (N( ⁇ )-eicosandioyl- ⁇ Glu-2xOEG), desB30 human insulin", “B29K(N ⁇ -eicosane diacyl- ⁇ Glu-2xOEG), desB30 human insulin", or "B29K (N ⁇ -eicosane diacyl-gGlu-2xOEG), desB30 human insulin” to indicate the amino acid K at position B29 in human insulin It has been modified by the acylation of the eicosandioyl-gGlu-2xOEG residue on the ⁇ nitrogen of the lysine residue of B29 (called N ⁇ or (N( ⁇ )), and the position B30 in human insulin T amino acid has been deleted.
  • insulin for Comparative example 5 is referred to as "A14E, B16H, B25H, B29K (N ⁇ eicosane diacyl -gGlu-2xOEG), desB30 Human insulin” or "A14E, B16H, B25H, B29K (N( ⁇ )-eicosanedioyl- ⁇ Glu-2xOEG), desB30 human insulin” means that the amino acid Y at position A14 in human insulin has been mutated to E, human The amino acid Y at position B16 in insulin has been mutated to H, the amino acid F at position B25 in human insulin has been mutated to H, and the amino acid K at position B29 in human insulin has passed the epsilon nitrogen (called It is modified by the acylation of eicosane diacyl-gGlu-2xOEG on N ⁇ ), and the amino acid T at position B30 in human insulin has been deleted.
  • A14E, B16H, B25H, B29K N ⁇
  • nxPEG represents the group -NH(CH 2 CH 2 O) n CH 2 CO-, where n is an integer.
  • 12xPEG represents the group -NH(CH 2 CH 2 O) 12 CH 2 CO-.
  • Insulin is a polypeptide hormone secreted by ⁇ -cells in the pancreas. It is composed of two polypeptide chains, A and B, which are connected by two interchain disulfide bonds. In addition, the A chain is characterized by an intra-chain disulfide bond.
  • the construction, expression, processing, and purification of the insulin analog vector can be carried out using techniques known to those skilled in the art.
  • the insulin analog can be prepared by a well-known technique disclosed in US Patent No. 6,500645 by expressing a DNA sequence encoding the target insulin analog in a suitable host cell.
  • insulin analogues can also be prepared by the method reported in the following literature: Glendorf T, AR, Nishimura E, Pettersson I, & Kjeldsen T: Importance of the Solvent-Exposed Residues of the Insulin B Chain ⁇ -Helix for Receptor Binding; Biochemistry 2008 47 4743-4751. This document uses overlap extension PCR to introduce mutations into the insulin encoding vector.
  • Insulin analogs are expressed in Saccharomyces cerevisiae strain MT663 as a pre-insulin-like fusion protein with Ala-Ala-Lys small C-peptide.
  • A. lyticus endoprotease the single-chain precursor is enzymatically converted into a double-chain desB30 analog.
  • the isolated insulin analogs can be acylated at desired positions by acylation methods known in the art. Examples of such insulin analogs have been described in, for example, Chinese patent applications with publication numbers CN1029977C, CN1043719A and CN1148984A.
  • each insulin analog polypeptide encoded can be synthesized by established standard methods, such as the method described in Beaucage et al. (1981) Tetrahedron Letters 22:1859-1869, or Matthes et al. (1984) EMBO Journal 3 : The method described in 801-805.
  • excipient in a broad sense refers to any component other than the active therapeutic ingredient.
  • the excipient may be an inert substance, an inactive substance and/or a non-pharmaceutically active substance.
  • Excipients can be used for various purposes, depending on the pharmaceutical composition, for example as a carrier, vehicle, diluent, tablet adjuvant, and/or for improving the administration and/or absorption of the active substance.
  • excipients include, but are not limited to, diluents, buffers, preservatives, tonicity regulators (also known as tonicity agents or isotonic agents), chelating agents, surfactants, protease inhibitors, wetting agents, emulsifiers , Antioxidants, fillers, metal ions, oily solvents, proteins and/or zwitterions and stabilizers.
  • compositions of pharmaceutically active ingredients and various excipients are known in the art, see, for example, Remington: The Science and Practice of Pharmacy (for example, the 19th edition (1995) and any later editions).
  • the patient prefers the time interval (time delay) from the administration of the acylated insulin of the present invention to the next administration of the acylated insulin of the present invention to have the same length or approximately the same length in days. It can even be expected that patients will prefer to administer acylated insulin once a week, that is, on the same day of the week, for example every Sunday. On average over a period of 1 month, 6 months, or 1 year, this would be acylated insulin administered every 6 days and not more frequently. For some patients, on average over a period of 1 month, 6 months, or 1 year, it may be necessary to administer acylated insulin every 5 days or approximately every 5 days and not more frequently.
  • acylated insulin For other patients, calculated on average over a period of 1 month, 6 months, or 1 year, it may be necessary to administer acylated insulin every 4 days or approximately every 4 days and not more frequently. For other patients, calculated on average over a period of 1 month, 6 months, or 1 year, it may be necessary to administer acylated insulin every 3 days or approximately every 3 days and not more frequently. Even other patients may find it advantageous to administer acyl groups on average over a period of 1 month, 6 months, or 1 year, twice a week, for example, at intervals of about 3-4 days between each administration. Insulin. For some patients, calculated on average over a period of 1 month, 6 months, or 1 year, it may be necessary to administer acylated insulin every 2 days or approximately every 2 days and not more frequently.
  • acylated insulin For other patients, calculated on average over a period of 1 month, 6 months, or 1 year, it may be necessary to administer acylated insulin every other day or approximately every other day and not more frequently. For some patients, calculated on average over a period of 1 month, 6 months, or 1 year, it may be necessary to administer acylated insulin every 7 days or approximately every 7 days and not more frequently. Even other patients may not administer acylated insulin at exactly the same length of time interval (in days) every week, month, or year. On average over a period of 1 month, 6 months, or 1 year, some patients may sometimes be given acylated insulin at intervals of 5 to 7 days and not at a higher frequency.
  • the diseases and conditions that are the main targets of the present invention are diabetes (type 1 or type 2) or other conditions characterized by hyperglycemia, but are generally metabolic diseases and conditions in which insulin metabolism is clinically relevant or beneficial , Such as pre-diabetes, impaired glucose tolerance, metabolic syndrome, obesity, cachexia, damage/death of ⁇ -cells in the body, excessive appetite, and inflammation. All these types of conditions are known or believed to benefit from the stable metabolic state of subjects suffering from the disease or condition.
  • any treatment regimen that includes administration of insulin can be modified by practicing the teachings of the present invention, meaning that such therapy will include administration of insulin according to the prolonged action provided herein.
  • OEG is the amino acid residue -NH(CH 2 ) 2 O(CH 2 ) 2 OCH 2 CO-
  • OSu is succinimidyl-1-yloxy-2,5-dioxo-pyrrolidin-1-yloxy
  • OtBu is oxy tert-butyl
  • HCl is hydrogen chloride
  • ⁇ Glu or gGlu is ⁇ L-glutamyl
  • NHS is N-hydroxysuccinimide
  • DCC is dicyclohexylcarbodiimide
  • AEEA is 2-(2-(2-aminoethoxy)ethoxy)acetic acid
  • CH 3 CN is acetonitrile
  • Gly is glycine
  • Arg is arginine
  • TFA is trifluoroacetic acid
  • HbA1c is glycosylated hemoglobin
  • AUC is the area under the curve of time-glucose curve
  • RU is the response unit (Response unit)
  • B29K N( ⁇ )-eicosandioyl- ⁇ Glu-5xOEG
  • desB30 human insulin compound 1
  • des(B30) human insulin was prepared according to the method described in Example 11 of Chinese Patent CN1056618C.
  • DesB30 human insulin (5 g, 0.876 mmol) was dissolved in 100 mM Na 2 HPO 4 aqueous solution (150 mL), and acetonitrile (100 mL) was added, and the pH was adjusted to pH 10-12.5 with 1 N NaOH.
  • Dissolve tert-butyl eicosane diacyl- ⁇ Glu-(5xOEG-OSu)-OtBu (1.36 g, 0.964 mmol) in acetonitrile (50 mL), and slowly add to the insulin solution. Maintain the pH at 10-12.5. After 120 minutes, the reaction mixture was added to water (150 mL), and the pH was adjusted to 5.0 with 1N aqueous HCl.
  • the precipitate was separated by centrifugation and lyophilized.
  • the crude product was added to a mixed solution of trifluoroacetic acid (60 mL) and dichloromethane (60 ml), and stirred at room temperature for 30 minutes.
  • the mixture was concentrated to about 30 ml, poured into ice-cold n-heptane (300 mL), and the precipitated product was separated by filtration and washed twice with n-heptane.
  • B29K N( ⁇ )-eicosandioyl- ⁇ Glu-6xOEG
  • desB30 human insulin compound 2
  • Compound 2 was prepared in a procedure similar to that of Example 1, Part 2.
  • B29K N( ⁇ )-eicosandioyl- ⁇ Glu-8xOEG
  • desB30 human insulin compound 3
  • B29K N( ⁇ )-docosanediacyl- ⁇ Glu-6xOEG
  • desB30 human insulin compound 4
  • Compound 4 was prepared in a procedure similar to that of Example 1, Part 2.
  • B29K N( ⁇ )-docosanedioyl- ⁇ Glu-8xOEG
  • desB30 human insulin compound 5
  • Compound 5 was prepared in a procedure similar to that of Example 1, Part 2.
  • the reference compound insulin degludec was prepared according to Example 4 of patent CN105820233A.
  • B29K N( ⁇ )-eicosandioyl- ⁇ Glu-2xOEG
  • desB30 human insulin control compound 2
  • control compound 2 was prepared in a procedure similar to that of Example 1 Part 2.
  • B29K N( ⁇ )-octadecane diacyl- ⁇ Glu-2xOEG
  • desB30 human insulin control compound 3
  • control compound 3 was prepared in a procedure similar to that of Example 1, Part 2.
  • B29K N( ⁇ )-octadecandioyl- ⁇ Glu-6xOEG
  • desB30 human insulin control compound 4
  • control compound 4 was prepared by a procedure similar to that of Example 1, Part 2.
  • the purpose of this study is to verify the acylated insulin of the present invention on the regulation of blood glucose (BG) in the case of diabetes.
  • acylated insulins of Examples 1-5 and the control compounds of Control Examples 1-4 were tested in a single-dose study.
  • the blood glucose lowering effect of the acylated insulin was tested at different doses of 9U/kg or 10U/kg.
  • mice 8-9 weeks old male db/db(BKS/Lepr) mice are raised in a barrier environment in a breeding box of suitable specifications, free access to standard food and purified water, and the environmental conditions are controlled at a relative humidity of 40%-60%.
  • the temperature is 22°C-24°C. After an adaptation period of 1-2 weeks, start to be used in experiments.
  • mice were weighed. According to random blood glucose and body weight, the mice were matched and assigned to the vehicle group or treatment group, and received the following treatment: subcutaneous injection of vehicle, or subcutaneous injection of acylated insulin 9U/kg or 10U/kg, where the vehicle contained: glycerol 19.6mg/ml, phenol 1.5mg/ml, m-cresol 1.72mg/ml, zinc ion concentration 55 ⁇ g/ml, pH value 7.6.
  • vehicle contained: glycerol 19.6mg/ml, phenol 1.5mg/ml, m-cresol 1.72mg/ml, zinc ion concentration 55 ⁇ g/ml, pH value 7.6.
  • the acylated insulin is dissolved in a solvent to an administration concentration of 1.8 or 2 U/ml, and the administration volume is 5 ml/kg (that is, 50 ⁇ l/10 g body weight).
  • Subcutaneous administration (S.C.) is adopted, and the drug is administered once subcutaneously on the back of the neck.
  • the acylated insulin was administered at approximately 10:30 in the morning (time 0).
  • time 0 time 0
  • the animals were fasted without water, and the blood glucose of the mice was evaluated 3, 6, 9, 12, and 15 hours after the administration.
  • the oral glucose tolerance test (OGTT) was started after the 15-hour blood glucose was detected in the test, and the blood glucose was measured 30min, 60min, 120min and 180min after intragastric administration of glucose solution (100mg/mL, 10mL/kg).
  • the OGTT experiment was carried out three times in a row. According to the results of the preliminary experiment, at the last OGTT experiment, the efficacy of the test compound was nearly disappeared, and it was terminated after 30 hours of blood glucose evaluation.
  • the area under the blood glucose-time curve (AUC) from 0 to the monitoring endpoint was calculated.
  • AUC blood glucose-time curve
  • Figures 1a and 1b show that the acylated insulins of the present invention, such as compound 1, and compound 2, have a significantly better hypoglycemic effect on db/db mice than insulin deglu, and the effective action time is prolonged compared with insulin deglu.
  • Figures 2a and 2b show that the acylated insulins of the present invention such as compound 1 and compound 2 have significantly better hypoglycemic effect on db/db mice than the control compound 2.
  • the compound 1 and compound 2 of the present invention are compared with the control compound 2.
  • the efficacy of the drug was increased by 39.5% and 45.1% in the interval of 0 to 16.5 hours of administration, as shown in Table 1:
  • Table 1 Increased efficacy of the acylated insulin of the present invention relative to the control compound 2
  • Percentage increase in efficacy relative to control compound 2 [(AUC(test compound)-AUC(vehicle))/((AUC(reference compound 2)-AUC(vehicle))-1]*100%, where test Compound refers to the acylated insulin of the present invention
  • Figures 3a-3b show that compound 1, compound 2 and compound 3 of the present invention have very good pharmacological effects, and in db/db mice, they are still effective after 30 hours of monitoring, and have a significantly prolonged hypoglycemic effect. .
  • Figures 4a-5b show that the acylated insulin of the present invention such as compound 2 has significantly better hypoglycemic effect on db/db mice than the control compound 3 and the control compound 4.
  • Figures 6a-6b show that compound 4, compound 5 and compound 2 of the present invention all have very good pharmacological effects, and in db/db mice, they are still effective after 41 hours of monitoring, and have a significantly prolonged hypoglycemic effect time .
  • mice Male wistar rats aged 8w weeks, weighing 180-220g. Raised in a barrier environment in a feeding box of suitable specifications (5 per box), free access to standard food and purified water, the environmental conditions are controlled at a relative humidity of 40% to 60%, and a temperature of 22°C to 24°C. After the 4-day adaptation period, fasting for 12 hours, rats were intraperitoneally injected with streptozotocin (sigma) solution (10 mg/mL in 0.1 M citrate buffer) at 60 mg/kg. After administration, to prevent sudden hypoglycemia in rats, appropriate supplementation of glucose (20%) in drinking water, and withdrawal of supplementation after 12 hours. After 4 days of administration of streptozotocin, random blood glucose detection was performed, and the rats with blood glucose values above 20 mmol/L were selected as T1DM model rats for subsequent experiments.
  • streptozotocin Sigma
  • the basal blood glucose was evaluated at time -1/1h (9:30 am), and the rats were weighed. According to random blood glucose and body weight, the rats were matched and assigned to the vehicle group or treatment group, and received the following treatment: subcutaneous injection of vehicle, or subcutaneous injection of acylated insulin, 3U/kg, where the vehicle contained: glycerol 19.6mg/ml, phenol 1.5mg/ ml, m-cresol 1.72 mg/ml, zinc ion concentration 55 ⁇ g/ml, and the pH of the solvent is 7.6.
  • vehicle contained: glycerol 19.6mg/ml, phenol 1.5mg/ ml, m-cresol 1.72 mg/ml, zinc ion concentration 55 ⁇ g/ml, and the pH of the solvent is 7.6.
  • the acylated insulin is dissolved in a solvent to an administration concentration of 1.5 U/ml, and the administration volume is 2 ml/kg (ie, 0.2 ml/100 g body weight). It is administered subcutaneously, once subcutaneously on the back of the neck.
  • the acylated insulin was administered at approximately 9:30 in the morning (time 0), and the blood glucose of the rats was evaluated at 2, 4 hours after the administration.
  • An oral glucose tolerance test (OGTT) test was performed at 4 hours and 7 hours respectively (see below for details).
  • Detection time At the specified time point, blood was collected from the tail tip to determine fasting blood glucose (0min), then glucose solution (100mg/mL or 200mg/mL, 10mL/kg) was given by gavage, and then the glucose was measured at 30min, 60min, 120min and 180min after the glucose load blood sugar.
  • a dose-response curve of blood glucose versus time is drawn.
  • AUC blood glucose-time curve
  • Figures 7a-7b show that the acylated insulin of the present invention also has a very good hypoglycemic effect in type I diabetes (T1DM) rats, that is, it has a very good pharmacological effect.
  • [Gly8, Arg34]GLP-1-(7-37) peptide is prepared by general protein recombinant expression method (for specific method, please refer to Molecular Cloning: A Laboratory Manual (Fourth Edition), Michael R. Green, Cold Spring Harbor Press, 2012) .
  • [Gly8,Arg34]GLP-1-(7-37) peptide (5g, 1.48mmol) was dissolved in 100mM Na 2 HPO 4 aqueous solution (150mL), and acetonitrile (100mL) was added, and the pH was adjusted to pH 10 with 1N NaOH -12.5.
  • the mixture was concentrated to about 30 ml, poured into ice-cold n-heptane (300 mL), the precipitated product was separated by filtration, and washed twice with n-heptane.
  • the product was purified by ion exchange chromatography (Ressource Q, 0.25%-1.25% ammonium acetate gradient in 42.5% ethanol, pH 7.5), reverse phase chromatography (acetonitrile, water, TFA) , The purified fractions were combined, the pH was adjusted to 5.2 with 1N HCl, the precipitate was separated, and lyophilized to obtain the title compound.
  • A14E, B16H, B25H, desB30 human insulin by conventional methods of preparing insulin analogs (see Glendorf T, AR, Nishimura E, Pettersson I, & Kjeldsen T: Importance of the Solvent-Exposed Residues of the Insulin B Chain ⁇ -Helix for Receptor Binding; Biochemistry 2008 47 4743-4751).
  • A14E, B16H, B25H, desB30 human insulin (5 g, 0.888 mmol) was dissolved in 100 mM Na 2 HPO 4 aqueous solution (150 mL), and acetonitrile (100 mL) was added, and the pH was adjusted to pH 10-12.5 with 1N NaOH.
  • the mixture was concentrated to about 30 ml, poured into ice-cold n-heptane (300 mL), and the precipitated product was separated by filtration and washed twice with n-heptane. After vacuum drying, it was purified by ion exchange chromatography ((Ressource Q, 0.25%-1.25% ammonium acetate gradient in 42.5% ethanol, pH 7.5), reverse phase chromatography (acetonitrile, water, TFA), and the purified The fractions were combined, the pH was adjusted to 5.2 with 1N HCl, and the precipitate was separated and lyophilized to obtain control compound 5.
  • ion exchange chromatography (Ressource Q, 0.25%-1.25% ammonium acetate gradient in 42.5% ethanol, pH 7.5), reverse phase chromatography (acetonitrile, water, TFA), and the purified
  • the fractions were combined, the pH was adjusted to 5.2 with 1N HCl, and the precipitate was separated and lyophilized to
  • A14E, B16H, B25H, B29K N( ⁇ )-eicosandioyl- ⁇ Glu-6xOEG), desB30 human insulin (compound 13)
  • A14E, B16H, B25H, B29K N( ⁇ )-docosane diacyl- ⁇ Glu-6xOEG), desB30 human insulin (compound 14)
  • the acylated insulin is dissolved in a solvent to a dosage of 33.5 U/ml, and the dosage is 1 ml/kg (that is, 0.1 ml/100 g body weight).
  • the acylated insulin was administered at approximately 9:30-10:00 (time 0) in the morning, and the blood glucose of rats was monitored at 3h, 6h, 9h, 24h, 48h, 72h, 96h, and 120h after administration.
  • acylated insulin for each single dose of acylated insulin (control compound 5, compound 14, compound 13), a dose response curve of blood glucose versus time was drawn. To illustrate the effect of acylated insulin on blood glucose, for each individual dose response curve, the area under the blood glucose-time curve (AUC) from 0 to the monitoring endpoint was calculated. Among them, the smaller the AUC value, the better the hypoglycemic effect and the better the drug effect.
  • Figures 8a-8b show that the acylated insulin of the present invention has unexpectedly increased pharmacodynamics.
  • the title compounds of Examples 15 and 16 of Compound 13 and Compound 14 induce STZ I Type 2 diabetes (T1DM) rats all have better hypoglycemic effect, that is, have better efficacy.
  • Example 17 With reference to the similar experimental procedure in Example 17, the pharmacodynamic study was carried out in streptozotocin (STZ)-induced type I diabetes (T1DM) female rats. The difference is that the acylated insulin is implemented
  • STZ streptozotocin
  • T1DM type I diabetes
  • Example 6 With reference to the similar experimental procedure of Example 6, on the obese diabetic mouse model (db/db mice), the title compound of Comparative Example 5, and the title compounds of Examples 15 and 16 (ie, the control compound) was tested in a single-dose study. 5. Compound 13 and Compound 14). The blood glucose lowering effect of the acylated insulin was tested at a dose of 9 U/kg.
  • mice 8-9 weeks old male db/db(BKS/Lepr) mice are raised in a barrier environment in a breeding box of suitable specifications, free access to standard food and purified water, and the environmental conditions are controlled at a relative humidity of 40%-60%.
  • the temperature is 22°C-24°C. After an adaptation period of 1-2 weeks, start to be used in experiments.
  • mice were weighed. According to random blood glucose and body weight, the mice were matched and assigned to the vehicle group or treatment group, and received the following treatment: subcutaneous injection of vehicle, or subcutaneous injection of acylated insulin 9U/kg, where the vehicle used contained: phenol 5.65mg/ml, glycerol 15mg/ml , Disodium hydrogen phosphate 0.708 mg/ml, sodium chloride 0.585 mg/ml, and the pH of the solvent is 7.6.
  • vehicle contained: phenol 5.65mg/ml, glycerol 15mg/ml , Disodium hydrogen phosphate 0.708 mg/ml, sodium chloride 0.585 mg/ml, and the pH of the solvent is 7.6.
  • the acylated insulin is dissolved in a solvent to an administration concentration of 1.8 U/ml, and the administration volume is 5 ml/kg (that is, 50 ⁇ l/10 g body weight).
  • Subcutaneous administration (S.C.) is adopted, and the drug is administered once subcutaneously on the back of the neck.
  • the acylated insulin was administered at approximately 10:30 in the morning (time 0).
  • time 0 time 0
  • the animals were fasted without water, and the blood glucose of the mice was evaluated 3h, 6h, 9h, and 21.5h after the administration.
  • the Oral Glucose Tolerance Test was started, and the blood glucose was measured 30min, 60min, 120min and 360min after intragastric administration of glucose solution (100mg/mL, 7.5mL/kg) ;
  • the second OGTT experiment was started after 330min of blood glucose was measured by the first OGTT, and the blood glucose was measured 30min, 90min, 210min and 360min after intragastric administration of glucose solution (50mg/mL, 10mL/kg).
  • the third OGTT experiment was started after the second OGTT measurement of blood glucose for 360 minutes.
  • the blood glucose was measured 30 minutes, 60 minutes, and 120 minutes after intragastric administration of glucose solution (50 mg/mL, 10 mL/kg).
  • glucose solution 50 mg/mL, 10 mL/kg.
  • the efficacy of the test compound Has not disappeared, the experiment was terminated after 36 hours of blood glucose assessment.
  • acylated insulin For each single dose of acylated insulin, a dose-response curve of blood glucose versus time is drawn. To illustrate the effect of acylated insulin on blood glucose, for each individual dose response curve, the area under the blood glucose-time curve (AUC) from 0 to the monitoring endpoint was calculated. Among them, the smaller the AUC value, the better the hypoglycemic effect and the better the drug effect.
  • Figures 10a-10b show that, compared to the control compound 5, the acylated insulin compound 14 and compound 13 of the present invention have significantly improved hypoglycemic effect in type 2 diabetic db/db mice.
  • the test was started 14 days after the model was built. Before the start of the experiment on the same day, the basal blood glucose was evaluated at time -1/1h (9:30 am), and the rats were weighed. According to random blood glucose and body weight, the rats were matched and assigned to the vehicle group or treatment group, and received the following treatment: subcutaneous injection of vehicle, or subcutaneous injection of the title compound of control example 5 and example 16 (ie control compound 5, compound 14), dose It is 25 U/kg, wherein the solvent contains: phenol 5.65 mg/ml, glycerol 15 mg/ml, disodium hydrogen phosphate 0.708 mg/ml, sodium chloride 0.585 mg/ml, and the pH of the solvent is 7.6.
  • the acylated insulin is dissolved in a solvent to an administration concentration of 25 U/ml, and the administration volume is 1 ml/kg (that is, 0.1 ml/100 g body weight).
  • Adopt subcutaneous administration method subcutaneous administration on the back of the neck, repeated administration 4 times, each time interval of 4 days, SD rats eat and drink freely during the experiment.
  • the acylated insulin was administered at approximately 9:30-10:00 (time 0) in the morning, and the blood glucose of rats was monitored after the first administration 3h, 6h, 9h, 24h, 48h, 72h, 96h, and 6h after each administration. The blood glucose of rats was monitored every 24h.
  • a dose-response curve of blood glucose versus time is drawn.
  • AUC blood glucose-time curve
  • the acylated insulin of the present invention has an unexpectedly increased hypoglycemic effect in type I diabetes (T1DM) rats after administration.
  • T1DM type I diabetes
  • the hypoglycemic effect of compound 14 The effect is significantly better than the control compound 5.
  • composition comprising the acylated insulin and insulin aspart of the present invention can regulate blood glucose (BG) in streptozotocin (STZ) induced type I diabetes (T1DM) C57/6J mice effect.
  • BG blood glucose
  • STZ streptozotocin
  • T1DM type I diabetes
  • mice 4-6 weeks old male C57/6J mice (purchased from Vitality) are raised in a barrier environment in a breeding box of appropriate specifications, free access to standard food and purified water, and the environmental conditions are controlled at a relative humidity of 40%- 60%, the temperature is 22°C-24°C. After an adaptation period of 1-2 weeks, start to be used in experiments.
  • mice were fasted for 12 hours, and the mice were intraperitoneally injected with streptozotocin (sigma) solution (10 mg/mL in 0.1 M citrate buffer) at 150 mg/kg. Three days after the administration of streptozotocin, random blood glucose testing was performed, and the T1DM model mice with blood glucose values above 20mmol/L were selected for follow-up experiments.
  • streptozotocin Sigma
  • mice were tested for random blood glucose and weighed. According to random blood glucose and body weight, mice were assigned to vehicle group or treatment group. There were 5 groups, 8 mice in each group. Each group received the following treatment: subcutaneous injection of vehicle, or subcutaneous injection of insulin aspart (0.36U/kg), or Subcutaneous injection of a pharmaceutical composition containing insulin deglubber and insulin aspart, wherein the injection doses of insulin deglubber and insulin aspart are respectively 0.84 U/kg and 0.36 U/kg, or subcutaneous injection containing the title compound of Example 4 of the present invention Two pharmaceutical compositions of compound 4 and insulin aspart, wherein when the two pharmaceutical compositions are injected, the injection doses of compound 4 are 0.82 U/kg and 0.64 U/kg, respectively, and the injection doses of insulin aspart are both 0.36 U/kg, wherein the solvent includes: glycerol 19.6 mg/ml, phenol 1.5 mg/ml, m-cresol 1.72 mg/ml, zinc ion concentration 55
  • the premix of compound 4 and insulin aspart was dissolved in the vehicle to an administration concentration of 0.072 U/mL (based on the concentration of insulin aspart in the premix), and the administration volume was 5 ml/kg (that is, 50 ⁇ l/10 g body weight).
  • Subcutaneous administration (S.C.) is adopted, and the drug is administered once subcutaneously on the back of the neck. The drug was administered at approximately 16:00 (time 0) in the afternoon.
  • the animals were fasted without water, and the blood glucose of the mice was evaluated at 0.5, 1, 2, 3, 6 and 15:00 after the administration.
  • Figures 12a-12b show that after administration of a pharmaceutical composition containing the acylated insulin and insulin aspart of the present invention, compared with the pharmaceutical composition containing deglubber and insulin aspart, in type I diabetes (T1DM) mice There is an unexpectedly increased hypoglycemic effect.
  • T1DM type I diabetes
  • composition containing the acylated insulin of the present invention and insulin aspart was tested against blood glucose in streptozotocin (STZ) induced type I diabetes (T1DM) C57/6J mice. BG)'s moderating effect.
  • STZ streptozotocin
  • T1DM induced type I diabetes
  • mice were tested for random blood glucose and weighed. According to random blood glucose and body weight, mice were assigned to vehicle group or treatment group. There were 7 groups, 8 mice in each group. Each group received the following treatment: subcutaneous injection of vehicle, or subcutaneous injection of insulin aspart (3U/kg), or subcutaneous Inject a pharmaceutical composition comprising insulin deglubber and insulin aspart, wherein the injection doses of insulin deglubber and insulin aspart are respectively 7 U/kg and 3 U/kg, or subcutaneously inject the title compound compound 4 and the title compound of Example 4 of the present invention.
  • subcutaneous injection of vehicle or subcutaneous injection of insulin aspart (3U/kg)
  • subcutaneous Inject a pharmaceutical composition comprising insulin deglubber and insulin aspart, wherein the injection doses of insulin deglubber and insulin aspart are respectively 7 U/kg and 3 U/kg, or subcutaneously inject the title compound compound 4 and the title compound of Example 4 of the present invention.
  • the injection doses of compound 4 are 6.79 U/kg, 5.34 U/kg, 3.84 U/kg and 2.39 U/kg, respectively.
  • the injection dose of winter insulin is 3U/kg
  • the solvent includes: glycerol 19.6mg/ml, phenol 1.5mg/ml, m-cresol 1.72mg/ml, zinc ion concentration 55 ⁇ g/ml, and the pH value of the solvent is 7.6.
  • the premix of compound 4 and insulin aspart was dissolved in a vehicle to a concentration of 0.6 U/mL (calculated as the concentration of insulin aspart in the premix), and the volume of administration was 5 ml/kg (that is, 50 ⁇ l/10 g body weight). It is administered by subcutaneous administration (S.C.) and subcutaneous injection on the back of the neck. The drug was administered at about 17:00 (time 0) every afternoon for 10 consecutive days. During the administration period, the mice were free to eat and drink. The blood glucose of the mice before the fourth, eighth, and tenth doses (0h) was evaluated, and the fourth, 8.
  • mice Random blood glucose of mice 1 hour after the tenth administration, random blood glucose before the eighth administration (0h) and 0.5, 1, 1.5, 2, 3, 4, 5, 6, 16, 24 hours after the administration Blood glucose is tested. After the last administration, the mice were fasted for 1 hour, and blood was taken from the orbit to detect the percentage of glycosylated hemoglobin (Hb1Ac) in the whole blood.
  • Hb1Ac glycosylated hemoglobin
  • Figures 13a-16 show that after administration of the composition containing the acylated insulin and insulin aspart of the present invention, compared with the pharmaceutical composition containing deglubber and insulin aspart, in type I diabetic (T1DM) mice It has an unexpectedly increased anti-diabetic effect, and has a more excellent anti-diabetic cumulative effect.
  • Figure 13a and Figure 13b show that a composition containing the acylated insulin and insulin aspart of the present invention, after administration, is in type I diabetes ( T1DM) mice have an unexpectedly increased hypoglycemic effect.
  • T1DM type I diabetes
  • Figures 14a-14c respectively show the blood glucose situation of mice in each administration group before the fourth, eighth, and tenth doses (0h), indicating that compared with the pharmaceutical composition containing deglubber and insulin aspart, the present invention is included
  • the pharmaceutical composition of acylated insulin and insulin aspart has better efficacy and a more excellent cumulative effect of lowering blood sugar.
  • Figures 15a-15c show the blood glucose situation of mice 1 hour after the fourth, eighth, and tenth doses, respectively, indicating that the acylated insulin of the present invention is contained relative to the pharmaceutical composition containing deglubber and insulin aspart
  • the pharmaceutical composition with insulin aspart has better medicinal effects and a more excellent cumulative effect of lowering blood sugar.
  • Figure 16 shows that the composition containing the acylated insulin and insulin aspart of the present invention has a better effect of reducing Hb1Ac after administration than the pharmaceutical composition containing insulin deglubber and insulin aspart.
  • the dosage ratio of insulin aspart is much smaller than the dosage ratio of insulin deglubber and insulin aspart, it can still achieve a better effect of reducing Hb1Ac.
  • the purpose of this experiment is to measure the chemical stability of the acylated insulin preparation of the present invention.
  • the title compound compound 4 of Example 4 was dissolved in 0.1% NaOH solution to a final concentration of 4.8 mM (pH value is about 10-11). According to the amount of each component in the table below, add phenol and m-cresol in sequence , Zinc acetate, glycerol and sodium chloride to produce an acylated insulin preparation with a final insulin concentration of 1.2mM (200U/ml or 8.46mg/ml), where the content of Zn is Zn/6 moles of acylated insulin (abbreviated as "Zn/6ins").
  • the chemical stability of the formulation in this example can be shown by the change in high molecular weight protein (HMWP) relative to day 0 after storage at 25°C and 37°C for 14 days and 20 days, and it can also be used at 25°C and 37°C.
  • HMWP high molecular weight protein
  • HMWP high molecular weight protein
  • High molecular weight protein content was determined by high performance liquid chromatography (HPLC), and the model specifications: Waters Xbride BEH 200A (7.8 * 300mm), 5 ⁇ m column, at a column temperature of 30 °C, temperature of the sample cell At 5°C, the test was performed with a mobile phase at a flow rate of 0.5 ml/min, where the mobile phase contained 600 ml of 0.1% arginine solution, 150 ml of glacial acetic acid and 250 ml of acetonitrile. The detection wavelength is 276nm, and the injection volume is 10ul. Table 2 shows the increase in HMWP on Day 14 and Day 20 relative to Day 0 at 25°C and 37°C.
  • the amount of HMWP in the above-mentioned acylated insulin preparations of the present invention increases very slowly with time, indicating that the above-mentioned acylated insulin preparations all have excellent chemical stability.
  • the Zn content is 6.5Zn/6ins
  • the amount of HMWP increases more slowly than when the Zn content is 5.5Zn/6ins.
  • the content of insulin-related substances was determined by high performance liquid chromatography (HPLC). On Waters Kromasil 300A-5 ⁇ m-C8 (4.6*250mm) column, when the column temperature is 40°C and the sample pool temperature is room temperature, the elution phase is used. The test was performed at a flow rate of 1.0 ml/min.
  • the elution phase consists of a mobile phase consisting of:
  • Phase A contains 0.1M anhydrous sodium sulfate, 0.1M sodium dihydrogen phosphate dihydrate, 10% acetonitrile (v/v), adjust the pH value to 5.0 with NaOH;
  • Phase B is 50% acetonitrile (v/v).
  • Table 3 shows the increase in related substances on the 14th and 20th day relative to the 0th day at 37°C.
  • the purpose of this experiment is to measure the chemical stability of the acylated insulin preparation of the present invention. According to the amounts of the components in the following Tables 4-6, following the steps similar to those in Example 23, the acylated insulin preparations in Table 4-6 were prepared. And following the procedure similar to that in Example 23, the changes of HMWP and related substances were measured. Tables 4-6 below show the changes of HMWP and related substances of acylated insulin preparations of different formulations.
  • the purpose of this experiment is to measure the chemical stability of the acylated insulin preparation of the present invention.
  • the acylated insulin preparations in Table 8 were prepared according to the same procedure as in Example 23.
  • the changes of HMWP and related substances were measured according to similar procedures as in Example 26.
  • the following table shows the changes of HMWP and related substances of acylated insulin preparations of different formulations.
  • the test was started 8 days after the model was built, and the basal blood glucose was monitored and the rats were weighed one day before the administration. According to random blood glucose and body weight, the rats were matched and assigned to the vehicle group or treatment group, and received the following treatment: subcutaneous injection of vehicle, or subcutaneous injection of insulin degludec (50U/kg), compound 4 (25U/kg or 40U/kg),
  • the solvent includes: phenol 60mM, glycerol 15mg/ml, m-cresol 10mM, sodium chloride 0.585mg/ml, and the pH value of the solvent is 7.4.
  • the acylated insulin is dissolved in a solvent to an administration concentration of 25 U/ml or 40 U/ml, and the administration volume is 1 ml/kg (ie, 0.1 ml/100 g body weight).
  • Adopt subcutaneous administration method subcutaneous administration on the back of the neck, once every other day, repeated administration 11 times, SD rats freely eat and drink during the experiment.
  • the acylated insulin was administered at approximately 9:30-10:30 in the morning.
  • the blood glucose of rats was monitored after the first administration of 3h, 4h, 5h, 6h, 24h, and 48h. After each administration, the rats’ blood glucose was monitored at 4h, 24h, and 48h. Rat blood sugar.
  • a dose-response curve of blood glucose versus time is drawn.
  • AUC blood glucose-time curve
  • the acylated insulin of the present invention has an unexpectedly increased hypoglycemic effect in type I diabetic (T1DM) rats after administration of insulin deglu, and the hypoglycemic effect of compound 4 Significantly better than insulin deglu.
  • composition containing the acylated insulin of the present invention and insulin aspart was tested against blood glucose in streptozotocin (STZ) induced type I diabetes (T1DM) C57/6J mice. BG)'s moderating effect.
  • STZ streptozotocin
  • T1DM induced type I diabetes
  • mice were weighed. According to random blood glucose and body weight, the mice were assigned to the vehicle group or treatment group. There were 8 groups, 9 mice in each group (5 males and 4 females).
  • a pharmaceutical composition of insulin glucomannan and insulin aspart wherein the injection doses of insulin degludec and insulin aspart are 7 U/kg and 3 U/kg, respectively, or subcutaneously injected containing the title compound compound 4 and insulin aspart of Example 4 of the present invention
  • the premix of compound 4 and insulin aspart was dissolved in a vehicle to a concentration of 0.6 U/mL (calculated as the concentration of insulin aspart in the premix), and the administration volume was 5 ml/kg (that is, 50 ⁇ l/10 g body weight). It is administered by subcutaneous administration (S.C.) and subcutaneous injection on the back of the neck. The drug was administered at about 16:00 (time 0) every afternoon for 15 consecutive days. During the administration period, the mice were free to eat and drink.
  • the first, second, fifth, eighth, and fifteenth doses were evaluated before (0h) and administration Random blood glucose of the mice after 1 hour, and the second, fifth, eighth, and fifteenth doses before (0h) blood glucose and 0.5, 1, 2, 4, 6, 16, 20, 24 hours after the administration of random blood glucose Detection. After the last administration, the mice were fasted for 2 hours, and blood was taken from the orbit to detect the percentage of glycosylated hemoglobin (Hb1Ac) in the whole blood.
  • Hb1Ac glycosylated hemoglobin
  • Figure 18a and Figure 18b show that the composition containing the acylated insulin and insulin aspart of the present invention, after administration, was compared with the pharmaceutical composition containing deglubber and insulin aspart in type I diabetic (T1DM) mice There is an unexpectedly increased hypoglycemic effect.
  • T1DM type I diabetic mice
  • Figure 19 shows that the composition comprising the acylated insulin and insulin aspart of the present invention has a better effect of reducing Hb1Ac compared to the pharmaceutical composition comprising insulin deglubber and insulin aspart after administration.
  • the dosage ratio of insulin aspart is much smaller than the dosage ratio of insulin deglubber and insulin aspart, it can still achieve a better effect of reducing Hb1Ac.
  • the purpose of this study is to verify the regulatory effect of the combination of the acylated insulin of the present invention and insulin aspart on blood glucose (BG) in an obese diabetic mouse model (db/db mice) in the case of diabetes.
  • mice 8-9 weeks old male db/db(BKS/Lepr) mice are raised in a barrier environment in a breeding box of suitable specifications, free access to standard food and purified water, and the environmental conditions are controlled at a relative humidity of 40%-60%.
  • the temperature is 22°C-24°C. After an adaptation period of 1-2 weeks, start to be used in experiments.
  • mice were tested for random blood glucose, and the mice were weighed. According to random blood glucose and body weight, mice were assigned to vehicle group or treatment group. There were 5 groups, 8 mice in each group. Each group received the following treatments: subcutaneous injection of vehicle or subcutaneous injection of a drug combination containing insulin degludec and insulin aspart Insulin, wherein the injection doses of insulin degludec and insulin aspart are respectively 7U/kg and 3U/kg, or subcutaneous injection of three pharmaceutical compositions containing the title compound compound 4 of Example 4 of the present invention and insulin aspart, wherein When the three pharmaceutical compositions were injected, the injection doses of compound 4 were 2.0 U/kg, 2.4 U/kg, 3.84 U/kg, and the injection doses of insulin aspart were all 3 U/kg, and the solvent contained: 60 mM phenol , 10mM m-cresol, 15mg/ml glycerol, 15mM Na 2 HPO 4 , pH value is 7.6.
  • the acylated insulin is dissolved in a solvent to an administration concentration of 0.6 U/ml, and the administration volume is 5 ml/kg (that is, 50 ⁇ l/10 g body weight).
  • Adopt subcutaneous administration (S.C.) method subcutaneous injection administration on the back of the neck four times.
  • the acylated insulin was administered at approximately 09:30 in the morning (time 0).
  • time 0 time 0
  • the animals were fasted without water, and the blood glucose of the mice was evaluated 0.5, 1, 2, 3, 4, 6, 8, 10, and 12 hours after the administration.
  • the area under the blood glucose-time curve (AUC) from 0 to the monitoring endpoint was calculated.
  • AUC blood glucose-time curve
  • Figures 20a and 20b show that the composition containing the acylated insulin and insulin aspart of the present invention, after administration, is compared with the pharmaceutical composition containing deglubber and insulin aspart in an obese diabetic mouse model ( db/db mice) has an unexpectedly increased hypoglycemic effect.
  • the dosage ratio of compound 4 to insulin aspart is much smaller than the dosage ratio of insulin degludec and insulin aspart, it can still achieve better lowering effects.
  • Sugar effect When the dosage ratio of compound 4 to insulin aspart is much smaller than the dosage ratio of insulin degludec and insulin aspart, it can still achieve better lowering effects.
  • Sugar effect When the dosage ratio of compound 4 to insulin aspart is much smaller than the dosage ratio of insulin degludec and insulin aspart, it can still achieve better lowering effects.
  • the purpose of this experiment is to measure the chemical stability of the acylated insulin preparation of the present invention.
  • the title compound compound 4 of Example 4 was dissolved in 0.03% NaOH solution to a concentration of 2.4 mM, and then the pH value was adjusted to 7.4 with 4% NaOH solution. According to the amount of each component in the following table, the phenol, m-methyl After mixing phenol, glycerin and sodium chloride, add to the compound 4 solution, adjust the pH to 7.4, and then add the zinc acetate to the compound 4 solution three times according to the amount of zinc acetate in the table below, and adjust the pH to the final value.
  • acylated insulin preparation with a final insulin concentration of 1.2 mM (200 U/ml or 8.46 mg/ml) was produced, where the content of Zn was expressed as Zn/6 moles of acylated insulin (abbreviated as "Zn/6ins").
  • the chemical stability of the formulation in this example can be shown by the change in high molecular weight protein (HMWP) relative to day 0 after storage at 25°C and 37°C for 14 days and 21 days, and it can also be used after storage at 37°C for 21 days.
  • HMWP high molecular weight protein
  • HMWP high molecular weight protein
  • HMWP high molecular weight protein
  • HPLC high performance liquid chromatography
  • the content of insulin-related substances was determined by high-performance liquid chromatography (HPLC). On a Waters Kromasil 100A-3.5 ⁇ m-C8 (4.6*250mm) column, the column temperature was 40°C and the sample pool temperature was 10°C. The dephasing is tested at a flow rate of 1.0 ml/min.
  • the elution phase consists of a mobile phase consisting of:
  • Phase A contains 0.1M anhydrous sodium sulfate, 0.1M sodium dihydrogen phosphate dihydrate, 10% acetonitrile (v/v), adjust the pH value to 3.0 with concentrated phosphoric acid;
  • Phase B is 60% acetonitrile (v/v).
  • the purpose of this experiment is to measure the chemical stability of the acylated insulin preparation of the present invention.
  • the acylated insulin preparations in Tables 10 and 11 were prepared according to the procedure similar to that in Example 29.
  • the changes of HMWP and related substances were measured according to similar procedures as in Example 29.
  • Tables 10 and 11 below show the changes of HMWP and related substances of acylated insulin preparations of different formulations.
  • the purpose of this experiment is to measure the chemical stability of the combined preparation of acylated insulin and insulin aspart of the present invention.
  • the chemical stability of the formulation in this example can be shown by the change of high molecular weight protein (HMWP) relative to day 0 after storage at 37°C for 14 days and 28 days.
  • HMWP high molecular weight protein
  • HMWP high molecular weight protein
  • HMWP high molecular weight protein
  • HPLC high performance liquid chromatography
  • the purpose of this experiment is to measure the chemical stability of the combined preparation of acylated insulin and insulin aspart of the present invention.
  • Combinations 6-10 were formulated according to the amounts of each component in Table 14 below. And the change of HMWP was measured according to the procedure similar to that in Example 31. Table 15 below shows the changes in HMWP of acylated insulin preparations of different formulations.
  • the purpose of this experiment is to measure the chemical stability of the combined preparation of acylated insulin and insulin aspart of the present invention.
  • Combinations 11 and 12 were formulated according to the amounts of each component in Table 16 below. And the change of HMWP was measured according to the procedure similar to that in Example 31. Table 17 below shows the changes in HMWP of acylated insulin preparations of different formulations.
  • the purpose of this study is to verify the effect of the composition comprising the acylated insulin and insulin aspart of the present invention on blood glucose (BG) in an obese diabetic mouse model (db/db mice) in the case of diabetes.
  • mice were tested for random blood glucose and the mice were weighed. According to random blood glucose and body weight, the mice were assigned to vehicle group or treatment group. There were 4 groups, 5 mice in each group.
  • the injections of the acylated insulin and insulin aspart are dissolved in a solvent to the corresponding administration concentration, and the administration volume is 5 ml/kg (that is, 50 ⁇ l/10 g body weight). It is administered subcutaneously (S.C.) once a day. During the administration period, the animals were free to eat and drink. The random blood glucose of mice was evaluated at 0.5, 1, 2, 3, 4, 6, and 8 hours after the 21st day of continuous administration and 0.5, 1, 2, after the 18th day of continuous administration. After 3, 4, 6, 8, and 10 hours, the mice were fasted for blood glucose.
  • Figures 21a to 21d show that the composition comprising the acylated insulin and insulin aspart of the present invention, after administration, is compared with the pharmaceutical composition comprising insulin deglubber and insulin aspart in an obese diabetic mouse model ( db/db mice) has an unexpectedly increased hypoglycemic effect.
  • the dosage ratio of compound 4 to insulin aspart is much smaller than the dosage ratio of insulin degludec and insulin aspart, it can still achieve better lowering effects. Glucose effect, and the hypoglycemic lasts longer.
  • C max peak concentration
  • T max peak time
  • T 1/2 terminal elimination half-life
  • AUC 0-t 0-t time-area under the blood glucose concentration time curve
  • AUC INF time from administration under the plasma concentration time curve Area to infinity
  • Vd apparent volume of distribution
  • Cl clearance rate
  • MRT average residence time
  • Compound 4 intravenous group and insulin degludec intravenous group were taken before administration and after administration 2min, 10min, 30min, 1h, 2h, 4h, 6h, 8h, 12h, 24h, 30h, 36h, 48h to determine the blood concentration of the compound.
  • the acylated insulin derivative compound 4 of the present invention exhibits a longer half-life and a more stable hypoglycemic effect in rats and beagle dogs.
  • B29K N( ⁇ )-docosanedioyl- ⁇ Glu-OEG), desB30 human insulin (compound 20)
  • the compound B29K (N( ⁇ )-docosanediacyl- ⁇ Glu-OEG), desB30 human insulin was prepared in a similar procedure to that of Example 1, Part 2.
  • the compound B29K (N( ⁇ )-docosane diacyl- ⁇ Glu-12xPEG), desB30 human insulin was prepared by the similar steps as in the second part of Example 1.
  • the purpose of this test is to prove the binding ability of the insulin derivative of the present invention to the insulin receptor.
  • the surface plasmon resonance (SPR) method was used to compare the compound 2 of the present invention and the control compound 2 in the absence of human serum albumin (HSA) and the presence of 2% HSA, respectively, with the extracellular domain of his-tagged insulin receptor A ( IRA, Sino Biological) binding ability test.
  • HSA human serum albumin
  • IRA his-tagged insulin receptor A
  • the samples were diluted with Running buffer (Cytiva) or 2.0% HSA-containing operating buffer, respectively, so that the injection concentration of compound 2 and control compound 2 samples were both 400 nM.
  • the NTA sensor chip (Cytiva) was selected, and the SPR analysis was performed on the Biacore T200 (Cytiva) at 25°C.
  • the insulin derivative sample to be tested was injected at a flow rate of 30 ⁇ L/min for 60 seconds and dissociated for 60 seconds.
  • 350mM EDTA (Cytiva) was injected at a flow rate of 10 ⁇ L/min for 60 seconds to regenerate the chip.
  • HBS-P buffer (Cytiva) was cleaned, the next sample could be tested.
  • the response value 4s before the dissociation of the sample is selected as the result of the binding force test to the receptor, and the test is repeated 3 times for each sample.
  • Figure 22 shows the receptor binding ability of Compound 2 and Control Compound 2 in the presence of 2% HSA (simulating physiological conditions) relative to 0% HSA. It can be seen from Figure 22 that in the presence of 2% HSA, compound 2 has a significantly improved receptor binding ability compared to the control compound 2. The effect of albumin on the receptor binding ability of the compound 2 of the present invention is significantly lower than that of the control. Compound 2.
  • the insulin derivatives of the present invention such as compound 2 have an unexpectedly significantly improved receptor binding capacity compared to the control compound 2, that is, the receptor binding ability of albumin to the insulin derivatives of the present invention
  • the effect of body binding capacity is significantly lower than that of the control compound 2.
  • A14E, B16H, B25H, desB30 human insulin B chain
  • A14E, B16E, B25H, desB30 human insulin B chain A14E, B16E, B25H, desB30 human insulin B chain:
  • A21G human insulin A chain A21G human insulin A chain:
  • A21G human insulin B chain A21G human insulin B chain:
  • A21G, desB30 human insulin A chain A21G, desB30 human insulin A chain:
  • A21G, desB30 human insulin B chain A21G, desB30 human insulin B chain:

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CN202080091196.3A CN114901682B (zh) 2019-12-30 2020-12-29 胰岛素衍生物
EP20910566.7A EP4086279A4 (en) 2019-12-30 2020-12-29 INSULIN DERIVATIVE
CN202410312576.5A CN118666989A (zh) 2019-12-30 2020-12-29 胰岛素衍生物
CN202410312489.XA CN118480114A (zh) 2019-12-30 2020-12-29 胰岛素衍生物
CA3166495A CA3166495A1 (en) 2019-12-30 2020-12-29 Insulin derivative
MX2022008174A MX2022008174A (es) 2019-12-30 2020-12-29 Derivado de insulina.
AU2020418205A AU2020418205A1 (en) 2019-12-30 2020-12-29 Insulin derivative
KR1020227024018A KR20220121833A (ko) 2019-12-30 2020-12-29 인슐린 유도체
CN202410254905.5A CN118420744A (zh) 2019-12-30 2020-12-29 胰岛素衍生物
US17/758,101 US20240239862A1 (en) 2019-12-30 2020-12-29 Insulin Derivative
JP2022540767A JP2023510206A (ja) 2019-12-30 2020-12-29 インスリン誘導体
BR112022013150A BR112022013150A2 (pt) 2019-12-30 2020-12-29 Derivado da insulina
US19/052,262 US20250228919A1 (en) 2019-12-30 2025-02-12 Insulin Derivative
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WO2022247773A1 (en) * 2021-05-24 2022-12-01 Sunshine Lake Pharma Co., Ltd. A novel acylated insulin analog
WO2022268208A1 (zh) * 2021-06-25 2022-12-29 甘李药业股份有限公司 含酰化胰岛素的药物组合物
WO2023093758A1 (zh) * 2021-11-24 2023-06-01 成都奥达生物科技有限公司 一种长效胰岛素类似物
CN116327890A (zh) * 2023-05-29 2023-06-27 北京先为达生物科技有限公司 口服递送的组合物及其应用
EP4086277A4 (en) * 2019-12-30 2024-02-14 Gan & Lee Pharmaceuticals Co., Ltd. Long-acting glp-1 compound
WO2024179606A1 (zh) * 2023-03-02 2024-09-06 甘李药业股份有限公司 一种glp-1化合物的医药用途
WO2025098502A1 (zh) * 2023-11-08 2025-05-15 甘李药业股份有限公司 包含胰岛素衍生物的药物组合物的治疗用途

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WO2022159395A1 (en) 2021-01-20 2022-07-28 Viking Therapeutics, Inc. Compositions and methods for the treatment of metabolic and liver disorders
MX2024003236A (es) 2021-09-15 2024-06-19 Viking Therapeutics Inc Composiciones y metodos para el tratamiento de trastornos metabolicos y hepaticos.
CN118574842A (zh) * 2022-01-28 2024-08-30 甘李药业股份有限公司 酰化胰岛素
EP4722236A1 (en) * 2023-05-24 2026-04-08 The United Bio-Technology (Hengqin) Co., Ltd. Insulin derivative and use thereof
CN116789801B (zh) * 2023-08-21 2023-11-14 南京赛诺生物技术有限公司 新型胰岛素衍生物及其用途
WO2025077751A1 (zh) * 2023-10-09 2025-04-17 甘李药业股份有限公司 Glp-1肽组合物及其用途
WO2025098457A1 (zh) * 2023-11-07 2025-05-15 甘李药业股份有限公司 用于降糖的酰化胰岛素
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EP4086277A4 (en) * 2019-12-30 2024-02-14 Gan & Lee Pharmaceuticals Co., Ltd. Long-acting glp-1 compound
WO2022247773A1 (en) * 2021-05-24 2022-12-01 Sunshine Lake Pharma Co., Ltd. A novel acylated insulin analog
WO2022268208A1 (zh) * 2021-06-25 2022-12-29 甘李药业股份有限公司 含酰化胰岛素的药物组合物
EP4361174A4 (en) * 2021-06-25 2025-11-19 Gan & Lee Pharmaceuticals Co Ltd ACYLATED PHARMACEUTICAL COMPOSITION CONTAINING INSULIN
WO2023093758A1 (zh) * 2021-11-24 2023-06-01 成都奥达生物科技有限公司 一种长效胰岛素类似物
WO2024179606A1 (zh) * 2023-03-02 2024-09-06 甘李药业股份有限公司 一种glp-1化合物的医药用途
CN116327890A (zh) * 2023-05-29 2023-06-27 北京先为达生物科技有限公司 口服递送的组合物及其应用
CN116327890B (zh) * 2023-05-29 2023-12-08 北京先为达生物科技有限公司 口服递送的组合物及其应用
WO2025098502A1 (zh) * 2023-11-08 2025-05-15 甘李药业股份有限公司 包含胰岛素衍生物的药物组合物的治疗用途

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