CN115337391A - Compound preparation of insulin compound and GLP-1 receptor agonist and medical application thereof - Google Patents

Compound preparation of insulin compound and GLP-1 receptor agonist and medical application thereof Download PDF

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CN115337391A
CN115337391A CN202210522343.9A CN202210522343A CN115337391A CN 115337391 A CN115337391 A CN 115337391A CN 202210522343 A CN202210522343 A CN 202210522343A CN 115337391 A CN115337391 A CN 115337391A
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insulin
pharmaceutical composition
lysine
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朱勇超
王淇
孙磊
张小洁
陈昊
杨晓容
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Jiangsu Hengrui Medicine Co Ltd
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Abstract

The disclosure relates to a compound preparation of an insulin compound and a GLP-1 receptor agonist and medical application thereof. Specifically, the present disclosure relates to a combination formulation comprising an insulin compound, a GLP-1 receptor agonist, and zinc ions, wherein: the content of zinc ions is 4-8 zinc ions per 6 molecules of the insulin compound. The disclosure also relates to a preparation method and medical application of the compound preparation.

Description

Compound preparation of insulin compound and GLP-1 receptor agonist and medical application thereof
Technical Field
The disclosure relates to the field of pharmaceutical compositions, and in particular relates to a compound preparation of an insulin compound and a GLP-1 receptor agonist and medical application thereof.
Background
The prevalence of diabetes is on the rapid rise and has become another important chronic non-infectious disease that seriously compromises resident health, following cardiovascular disease and tumors. Disability and lethality caused by diabetes and various acute and chronic complications thereof have become the third world diseases threatening human health, and seriously affecting the quality of life of patients.
Insulin is a polypeptide consisting of 51 amino acids, which are distributed over 2 amino acid chains: an A chain (with 21 amino acids) and a B chain (with 30 amino acids). The chains are connected together by 2 disulfide bridges. Insulin preparations have been used for the treatment of diabetes for many years. WO2018024186A1 describes an acylated derivative of a human insulin analogue, a basal insulin with a long duration of action and an initial effect that is a gentle hypoglycemic effect.
Glucagon-like peptide 1 (GLP-1) is an endocrine hormone that increases insulin response upon oral ingestion of glucose or fat. GLP-1 generally regulates glucagon concentration, slows gastric emptying, stimulates insulin (pro) biosynthesis, increases sensitivity to insulin, and stimulates insulin-independent glycogen biosynthesis. CN101987868B describes a short-acting GLP-1 analogue, which is an artificially synthesized GLP-1 derivative containing a palmitic acid structure, and a main peptide chain consists of 40 amino acids. Has high development value and has a structure of HOOC (CH) 2 ) 14 -the CO-lipophilic substituent and the alpha amino group or epsilon amino group of the C-terminal Lys of the following amino acid sequence are linked by means of an amide bond:
His-Aib-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Glu-Glu-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-Leu-Val-Arg-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-Lys。
CN101861333A describes a compound formulation of insulin analogue and liraglutide, which needs to increase the concentration of zinc ion and give insulin analogue and zinc ion sufficient action time during formulation process because liraglutide competes with insulin for zinc ion.
Due to the variety of stability and activity of different polypeptide analogs, there is still a need in the art to develop a combination formulation of insulin and a GLP-1 analog with superior stability and activity.
Disclosure of Invention
The present disclosure provides a pharmaceutical composition comprising an insulin compound, a GLP-1 receptor agonist and zinc ions, wherein the amount of zinc ions is 4-8 zinc ions per 6 molecules of the insulin compound, the GLP-1 receptor agonist having a structure as shown and having a general structural formula R 1 (CH 2 ) n -lipophilic substituent of CO-:
X 1 -X 2 -Glu-Gly-Thr-Phe-Thr-Ser-Asp-X 10 -Ser-X 12 -X 13 -X 14 -Glu-X 16 -X 17 -Ala-X 19 -X 20- X 21 -Phe-Ile-X 24 -Trp-Leu-X 27 -X 28 -X 29 -X 30 -X 31 -X 32 -X 33 -X 34 -X 35 -X 36 -X 37 -X 38 -X 39 -Lys wherein R 1 Is selected from CH 3 -or HOOC-, n is an integer between 8 and 25, X 1 、X 2 、X 10 、X 12 、X 13 、X 14 、X 16 、X 17 、X 19 、X 20 、X 21 、X 24 、X 27 、X 28 、X 29 、X 30 、X 31 、X 32 、X 33 、X 34 、X 35 、X 36 、X 37 、X 38 、X 39 Each independently is selected fromNatural amino acids or unnatural amino acids or peptide fragments composed thereof are meant.
In some embodiments, the general structural formula is R 1 (CH 2 ) n -the lipophilic substituent of CO-and the amino group of the amino acid residue of the GLP-1 analogue are linked by means of an amide bond, wherein R 1 Is selected from CH 3 -or HOOC-, n is an integer from 8 to 25.
In some embodiments, the general structural formula is R 1 (CH 2 ) n -the lipophilic substituent of CO-and the epsilon amino group of the C-terminal Lys of the GLP-1 analog are linked by means of an amide bond, wherein R is 1 Is selected from CH 3 -or HOOC-, n is an integer from 8 to 25.
In some embodiments, the general structural formula is R 1 (CH 2 ) n -the lipophilic substituent of CO-and the alpha amino group of the C-terminal Lys of the GLP-1 analogue are linked by means of an amide bond, wherein R is 1 Is selected from CH 3 -or HOOC-, n is an integer from 8 to 25, for example 14.
In some embodiments, X in the amino acid sequence of the GLP-1 analog 1 Selected from L-His, D-His; x 2 Selected from Ala, D-Ala, gly, val, leu, ile, lys, aib; x 10 Selected from Val, leu; x 12 Selected from Ser, lys, arg; x 13 Selected from Tyr, gln; x 14 Selected from Leu, met; x 16 Selected from Gly, glu and Aib; x 17 Selected from Gln, glu, lys, arg; x 19 Selected from Ala, val; x 20 Selected from Lys, glu, arg; x 21 Selected from Glu, leu; x 24 Selected from Val, lys; x 27 Selected from Val, lys; x 28 Selected from Lys, glu, asn, arg; x29 is selected from Gly, aib; x 30 Selected from Arg, gly, lys; x 31 Selected from Gly, ala, glu, pro, lys; x 32 Selected from Lys, ser; x 33 Selected from Lys, ser; x 34 Selected from Gly, ala and Sar; x 35 Selected from Gly, ala and Sar; x 36 Is selected from Pro, gly; x 37 Is selected from Pro, gly; x38 is selected from Pro, gly; x 39 Is selected from Ser and Tyr.
The present disclosure provides a pharmaceutical composition comprising an insulin compound, a GLP-1 receptor agonist and zinc ions, wherein the content of zinc ions is 4-8 zinc ions per 6 molecules of the insulin compound, the GLP-1 receptor agonist having a structure as shown in formula (I):
Figure BDA0003642159730000031
in some embodiments, the amount of zinc ions is 4, 5, 6, 7, or 8 zinc ions per 6 molecules of the insulin compound, for example 5 zinc ions per 6 molecules of the insulin compound.
In some embodiments, the amount of zinc ion is 4-7 zinc ions, 4-6 zinc ions, or 5-7 zinc ions per 6 molecules of the insulin compound.
<xnotran> , pH 6.5-8.5, 6.6-8.5, 6.7-8.5, 6.8-8.5, 6.9-8.5, 7.0-8.5, 7.1-8.5, 7.2-8.5, 7.3-8.5, 7.4-8.5, 7.5-8.5, 7.6-8.5, 7.7-8.5, 7.8-8.5, 7.9-8.5, 8.0-8.5, 8.1-8.5, 8.2-8.5, 8.3-8.5, 8.4-8.5, 6.5-8.4, 6.6-8.4, 6.7-8.4, 6.8-8.4, 6.9-8.4, 7.0-8.4, 7.1-8.4, 7.2-8.4, 7.3-8.4, 7.4-8.4, 7.5-8.4, 7.6-8.4, 7.7-8.4, 7.8-8.4, 7.9-8.4, 8.0-8.4, 8.1-8.4, 8.2-8.4, 8.3-8.4, 6.5-8.3, 6.6-8.3, 6.7-8.3, 6.8-8.3, 6.9-8.3, 7.0-8.3, 7.1-8.3, 7.2-8.3, 7.3-8.3, 7.4-8.3, 7.5-8.3, 7.6-8.3, 7.7-8.3, 7.8-8.3, 7.9-8.3, 8.0-8.3, 8.1-8.3, 8.2-8.3, 6.5-8.2, 6.6-8.2, 6.7-8.2, 6.8-8.2, 6.9-8.2, 7.0-8.2, 7.1-8.2, 7.2-8.2, 7.3-8.2, 7.4-8.2, 7.5-8.2, 7.6-8.2, 7.7-8.2, 7.8-8.2, 7.9-8.2, 8.0-8.2, 8.1-8.2, 6.5-8.1, 6.6-8.1, 6.7-8.1, 6.8-8.1, 6.9-8.1, 7.0-8.1, 7.1-8.1, 7.2-8.1, 7.3-8.1, </xnotran> 7.4-8.1, 7.5-8.1, 7.6-8.1, 7.7-8.1, 7.8-8.1, 7.9-8.1, 8.0-8.1, 6.5-8.0, 6.6-8.0, 6.7-8.0, 6.8-8.0, 6.9-8.0, 7.0-8.0, 7.1-8.0, 7.2-8.0, 7.3-8.0, 7.4-8.0, 7.5-8.0, 7.6-8.0, 7.7-8.0, 7.8-8.0, 7.9-8.0 6.5-7.9, 6.6-7.9, 6.7-7.9, 6.8-7.9, 6.9-7.9, 7.0-7.9, 7.1-7.9, 7.2-7.9, 7.3-7.9, 7.4-7.9, 7.5-7.9, 7.6-7.9, 7.7-7.9, 7.8-7.9, 6.5-7.8, 6.6-7.8, 6.7-7.8, 6.8-7.8, 6.9-7.8, 7.0-7.8, 7.1-7.8 7.2-7.8, 7.3-7.8, 7.4-7.8, 7.5-7.8, 7.6-7.8, 7.7-7.8, 6.5-7.7, 6.6-7.7, 6.7-7.7, 6.8-7.7, 6.9-7.7, 7.0-7.7, 7.1-7.7, 7.2-7.7, 7.3-7.8, 7.4-7.7, 7.5-7.7, 7.6-7.7, 6.5-7.6, 6.6-7.6, 6.7-7.6 6.8-7.6, 6.9-7.6, 7.0-7.6, 7.1-7.6, 7.2-7.6, 7.3-7.6, 7.4-7.6, 7.5-7.6, 6.5-7.5, 6.6-7.5, 6.7-7.5, 6.8-7.5, 6.9-7.5, 7.0-7.5, 7.1-7.5, 7.2-7.5, 7.3-7.5, 7.4-7.5, 6.5-7.4, 6.6-7.4, 6.7-7.4, 6.8-7.4, 6.9-7.4, 7.0-7.4, 7.1-7.4, 7.2-7.4, 7.3-7.4, 6.5-7.3, 6.6-7.3, 6.7-7.3, 6.8-7.3, 6.9-7.3, 7.0-7.3, 7.1-7.3, 7.2-7.3, 6.5-7.2, 6.6-7.2, 6.7-7.2, 6.8-7.2, 6.9-7.2, 7.0-7.2, 7.1-7.2, 6.5-7.1 6.6-7.1, 6.7-7.1, 6.8-7.1, 6.9-7.1, 7.0-7.1, 6.5-7.0, 6.6-7.0, 6.7-7.0, 6.8-7.0, 6.9-7.0, 6.5-6.9, 6.6-6.9, 6.7-6.9, 6.8-6.9, 6.5-6.8, 6.6-6.8, 6.7-6.8, 6.5-6.7, 6.6-6.7, or 6.5-6.6.
In some embodiments, the pH of the pharmaceutical composition is 7.0-8.0, e.g., 7.3-7.8.
In some embodiments, the pH of the pharmaceutical composition is about 6.5, about 6.6, about 6.7, about 6.8, about 6.9, about 7.0, about 7.1, about 7.2, about 7.3, about 7.4, about 7.5, about 7.6, about 7.7, about 7.8, about 7.9, about 8.0, about 8.1, about 8.2, about 8.3, about 8.4, about 8.5, e.g., about 7.5.
In some embodiments, the pharmaceutical composition further comprises: at least one of a stabilizer, an osmotic pressure regulator, a preservative and a pH regulator.
In some embodiments, the stabilizing agent is selected from one or more of zinc acetate, zinc chloride, zinc sulfate, and zinc gluconate.
In some embodiments, the tonicity modifier is selected from mannitol, sorbitol, lactose, dextrose, trehalose, sodium chloride, glycerol, or any mixture thereof.
In some embodiments, the preservative is selected from phenol, cresol, m-cresol, paraben, or any mixture thereof.
In some embodiments, the pH adjusting agent is selected from one or more of hydrochloric acid, acetic acid, sulfuric acid, phosphoric acid, sodium hydroxide, TRIS, carbonate, and phosphate.
In some embodiments, the pharmaceutical composition comprises an insulin compound, a GLP-1 receptor agonist, glycerol, phenol, zinc acetate.
In some embodiments, the insulin compound is selected from the group consisting of native insulin, insulin analogs, and insulin derivatives.
In some embodiments, the native insulin is selected from human insulin, bovine insulin, and porcine insulin.
In some embodiments, an insulin analogue comprises less than 10 amino acid modifications (substitutions, deletions, additions (including insertions), and any combination thereof) relative to human insulin, or less than 9, 8, 7, 6, 5, 4, 3, 2, or 1 modification relative to human insulin. Examples of insulin analogs include, but are not limited to: des (B30) -human insulin; asp (B28) -human insulin; asp (B28) des (B30) -human insulin; lys (B3) Glu (B29) -human insulin; lys (B28) Pro (B29) -human insulin; gly (a 21) Arg (B31) Arg (B32) -human insulin; glu (a 14) His (B25) -human insulin; his (a 14) His (B25) -human insulin; glu (A14) His (B25) des (B30) -human insulin; his (A14) His (B25) des (B30) -human insulin; glu (A14) His (B25) des (B27) des (B28) des (B29) des (B30) -human insulin; glu (a 14) His (B25) Glu (B27) des (B30) -human insulin; glu (a 14) His (B16) His (B25) des (B30) -human insulin; his (a 14) His (B16) His (B25) des (B30) -human insulin; his (A8) Glu (A14) His (B25) Glu (B27) des (B30) -human insulin; his (A8) Glu (a 14) Glu (B1) Glu (B16) His (B25) Glu (B27) des (B30) -human insulin; and His (A8) Glu (a 14) Glu (B16) His (B25) des (B30) -human insulin.
In some embodiments, the insulin analogue is selected from Gly (a 21) Arg (B31) Arg (B32) -human insulin, lys (B3) Glu (B29) -human insulin, asp (B28) -human insulin, lys (B28) Pro (B29) -human insulin, des (B30) -human insulin.
In some embodiments, the insulin derivative is selected from insulin deglutamide, insulin detemir and compounds having the structure shown in formula (II) below:
S-W-X-Y-Z
(II)
wherein S is human insulin with 30 th threonine deletion of B chain; -W-X-Y-Z is an acylated modification group of an insulin analogue;
wherein W is a compound having-OC (CH) 2 ) n A diacyl structure of CO-, wherein N is an integer between 2 and 10, which forms an amide bond with an α -amino group of an a-chain or B-chain N-terminal amino acid residue of the parent insulin or an analog thereof or an ∈ -amino group of a lysine residue present on the B-chain with one of its acyl groups;
x is a diamino compound containing a carboxylic acid group, which is connected with one of the amino groups of the compound and one of the acyl groups in W to form an amide bond;
y is-A (CH) 2 ) m -wherein m is an integer from 6 to 32, such as an integer from 10 to 16, such as an integer from 12 to 14, a is absent or is CO-;
z is-COOH.
In some embodiments, the insulin derivative has the structure shown in formula (II) below:
S-W-X-Y-Z
(II)
wherein S is human insulin with 30 th threonine deletion of B chain; -W-X-Y-Z is an acylated modification group of an insulin analogue;
w forms an amide bond with an epsilon-amino group of a lysine residue present on the B-chain;
n is an integer from 2 to 5, such as 2; and
x is-HN (CH) 2 ) pCH (COOH) NH-, p is an integer from 2 to 10, such as an integer from 2 to 6, such as an integer from 2 to 4, such as 4.
In some embodiments, the insulin derivative is lysine B29 (N) ε -(N α -hexadecane fatty diacid-L-lysine-N ε -oxobutanoyl)) des (B30) human insulin.
In some embodiments, the pharmaceutical composition comprises: lysine B29 (N) ε -(N α -hexadecane fatty diacid-L-lysine-N ε -oxobutanoyl)) des (B30) human insulin, GLP-1 receptor agonist and zinc ion, wherein the content of zinc ion is4-8 zinc ions per 6 insulin compound molecules.
In some embodiments, the pharmaceutical composition comprises: lysine B29 (N) ε -(N α -hexadecane fatty diacid-L-lysine-N ε -oxobutanoyl)) des (B30) human insulin, a GLP-1 receptor agonist and zinc ions, wherein the content of zinc ions is 5-7 zinc ions per 6 molecules of the insulin compound.
In some embodiments, the pharmaceutical composition comprises: lysine B29 (N) ε -(N α -hexadecane fatty diacid-L-lysine-N ε -oxobutanoyl)) des (B30) human insulin, GLP-1 receptor agonist and zinc ion, wherein the content of zinc ion is 5 zinc ions per 6 insulin compound molecules.
In some embodiments, the pharmaceutical composition comprises: lysine B29 (N) ε -(N α -hexadecane fatty diacid-L-lysine-N ε -oxobutanoyl)) des (B30) human insulin, a GLP-1 receptor agonist, zinc ions, glycerol and phenol, wherein the content of zinc ions is 4-8 zinc ions per 6 molecules of the insulin compound, such as 5-7 zinc ions per 6 molecules of the insulin compound, such as 5 zinc ions per 6 molecules of the insulin compound.
The present disclosure provides a pharmaceutical composition comprising: human insulin, GLP-1 receptor agonist, and zinc acetate; optionally, further comprising glycerol and/or phenol. The GLP-1 receptor agonist has a structure shown in a formula (I). Such as lysine B29 (N) ε -(N α -hexadecane fatty diacid-L-lysine-N ε -oxobutanoyl)) des (B30) human insulin.
In some embodiments, the pharmaceutical composition comprises:
0.3-1.2mmol/L lysine B29 (N) ε -(N α -hexadecane fatty diacid-L-lysine-N ε -oxobutanoyl)) des (B30) human insulin,
0.1-1mg/mLGLP-1 receptor agonist, and
0.1-1.2mmol/L zinc acetate;
optionally, further comprising 10-30mg/mL glycerol, and/or 1-10mg/mL phenol;
in some embodiments, the pharmaceutical composition comprises:
0.4-0.8mmol/L lysine B29 (N) ε -(N α -hexadecane fatty diacid-L-lysine-N ε -oxobutanoyl)) des (B30) human insulin,
0.24-0.4mg/mLGLP-1 receptor agonist, and
0.4-0.8mmol/L zinc acetate;
optionally, further comprising 10-30mg/mL glycerol, and/or 1-10mg/mL phenol;
the pH of the pharmaceutical composition is 7.0-8.0, preferably 7.3-7.8.
In some embodiments, the pharmaceutical composition comprises:
0.6mmol/L lysine B29 (N) ε -(N α -hexadecane fatty diacid-L-lysine-N ε -oxobutanoyl)) des (B30) human insulin,
0.24-0.4mg/mLGLP-1 receptor agonist, and
0.5mmol/L of zinc acetate,
optionally, further comprising 19.7mg/mL of glycerol, and/or 5.7mg/mL of phenol,
the pH of the pharmaceutical composition is 7.0-8.0, preferably 7.3-7.8.
In some embodiments, the pharmaceutical composition comprises:
0.6mmol/L lysine B29 (N) ε -(N α -hexadecane fatty diacid-L-lysine-N ε -oxobutanoyl)) des (B30) human insulin,
0.24mg/mL GLP-1 receptor agonist,
0.5mmol/L of zinc acetate,
19.7mg/mL of glycerol, and
5.7mg/mL of phenol,
the pH of the pharmaceutical composition is 7.0-8.0, preferably 7.3-7.8.
In some embodiments, the pharmaceutical composition comprises:
0.6mmol/L lysine B29 (N) ε -(N α -hexadecane fatty diacid-L-lysine-N ε -oxobutanoyl)) des (B30) human insulin,
0.32mg/mL GLP-1 receptor agonist,
0.5mmol/L of zinc acetate,
19.7mg/mL of glycerol, and
5.7mg/mL of phenol,
the pH of the pharmaceutical composition is 7.0-8.0, preferably 7.3-7.8.
In some embodiments, the pharmaceutical composition comprises:
0.6mmol/L lysine B29 (N) ε -(N α -hexadecane fatty diacid-L-lysine-N ε -oxobutanoyl)) des (B30) human insulin,
0.4mg/mL GLP-1 receptor agonist,
0.5mmol/L of zinc acetate,
19.7mg/mL of glycerol, and
5.7mg/mL of phenol,
the pH of the pharmaceutical composition is 7.0-8.0, preferably 7.3-7.8.
In some embodiments, the pH of the above pharmaceutical composition may be about 7.5.
In some embodiments, the pharmaceutical composition is a soluble pharmaceutical composition for parenteral administration, for example, as an injection, a lyophilized formulation, or a solution resulting from reconstitution of the lyophilized formulation. In some embodiments, the pharmaceutical composition is an injection.
The present disclosure also provides a pharmaceutical composition comprising an insulin compound that is lysine B29 (N) and a GLP-1 receptor agonist ε -(N α -hexadecane fatty diacid-L-lysine-N ε -oxobutanoyl)) des (B30) human insulin, said GLP-1 receptor agonist having a structure according to formula (I):
Figure BDA0003642159730000071
optionally, the pharmaceutical composition further comprises zinc acetate.
In some embodiments, the GLP-1 receptor agonist does not compete with the insulin compound for zinc ions in the pharmaceutical compositions of the present disclosure.
In some embodiments, the pharmaceutical compositions of the present disclosure are stable at 2-8 ℃ for at least 3 months, at least 6 months, at least 12 months, at least 18 months, or at least 24 months.
In some embodiments, the pharmaceutical compositions of the present disclosure are stable for at least 7 days, at least 14 days, at least 28 days, at least 30 days, at least 1 month, at least 2 months, at least 3 months, at least 6 months, at least 12 months, at least 18 months, or at least 24 months at 25 ℃ or room temperature.
In some embodiments, the pharmaceutical composition of the present disclosure has a maximum single impurity of less than 0.90%, 0.85%, 0.80%, 0.75%, 0.70%, 0.65%, 0.60%, 0.55%, or 0.50% by mass of the insulin compound under 30-day standing conditions at 40 ℃; a21 deamidation is less than 1.5%, 1.4%, 1.3%, 1.2%, 1.1%, 1.0%, 0.9%, 0.8%, 0.7%, 0.6%, 0.5%, 0.4%, or 0.3%; b3 deamination is less than 0.5%, 0.45%, 0.4%, 0.35%, 0.3%, 0.25%, 0.2%, 0.15%, or 0.1%; and/or less than 5.5%, 5.4%, 5.3%, 5.2%, 5.1%, 5.0%, 4.9%, 4.8%, 4.7%, 4.6%, 4.5%, 4.4%, 4.3%, 4.2%, 4.1%, 4.0%, 3.9%, 3.8%, 3.7%, 3.6%, 3.5%, 3.4%, 3.3%, 3.2%, 3.1%, or 3.0% total impurities.
In some embodiments, the pharmaceutical composition of the present disclosure has a maximum single impurity of the GLP-1 receptor agonist of less than 1.5%, 1.4%, 1.3%, 1.2%, 1.1%, 1.0%, 0.9%, or 0.8% by mass under 30 day standing conditions at 40 ℃; the total impurities are less than 6.5%, 6.0%, 5.5%, 5.0%, 4.9%, 4.8%, 4.7%, 4.6%, 4.5%, 4.4%, 4.3%, 4.2%, 4.1%, or 4.0%.
In some embodiments, the pharmaceutical composition of the present disclosure does not exceed 0.6%, 0.55%, 0.5%, 0.45%, 0.4%, 0.35%, or 0.3% by mass of the high molecular protein under 30-day standing conditions at 40 ℃.
In some embodiments, the pharmaceutical composition of the present disclosure has a maximum single impurity of less than 0.80%, 0.75%, 0.70%, 0.65%, 0.60%, 0.55%, or 0.50% by mass of the insulin compound under 30-day standing conditions at 25 ℃; a21 deamidation is less than 0.5%, 0.4%, 0.3%, 0.2%, or 0.1%; b3 deamination is less than 0.4%, 0.35%, 0.3%, 0.25%, 0.2%, 0.15%, or 0.1%; and/or less than 3.0%, 2.5%, 2.0%, 1.5%, or 1.0% total impurities.
In some embodiments, the pharmaceutical composition of the present disclosure has a maximum single impurity of the GLP-1 receptor agonist of less than 0.8%, 0.7%, 0.6%, 0.5%, or 0.4% by mass under 30 day standing conditions at 25 ℃; the total impurities are less than 3.5%, 3.4%, 3.3%, 3.2%, 3.1%, or 3.0%.
In some embodiments, the pharmaceutical composition of the present disclosure does not exceed 0.35%, 0.3%, 0.25%, or 0.2% by mass of the high molecular protein under the condition of 30 days standing at 25 ℃.
The present disclosure also provides a method for preparing the above pharmaceutical composition, which comprises:
(1) Dissolving insulin compound in 5-15mM NaOH solution, adding a stabilizer for providing zinc ions, and adjusting pH to obtain insulin compound mother liquor;
(2) Adding a GLP-1 receptor agonist into a 5-15mM NaOH solution, and adjusting the pH value to obtain a GLP-1 receptor agonist mother solution;
(3) The insulin compound mother liquor is mixed with the GLP-1 receptor agonist mother liquor, and the pH is adjusted.
In some embodiments, there need not be a time interval, e.g., 48 hours, 24 hours, 12 hours, 6 hours, 4 hours, or 2 hours, before the mother liquor of insulin compound is mixed with the mother liquor of GLP-1 receptor agonist.
In some embodiments, no additional zinc ions, i.e., zinc ions other than the prescribed amount, need to be added during the formulation of the insulin compound master solution.
The present disclosure also provides a combination or product comprising a GLP-1 receptor agonist of formula (I) and lysine B29 (N) ε -(N α -hexadecane fatty diacid-L-lysine-N ε -oxobutanoyl)) des (B30) humanInsulin, optionally, the combination or product further comprises zinc ions, such as zinc acetate.
The present disclosure also provides a kit comprising a first pharmaceutical composition comprising a GLP-1 receptor agonist of formula (I) and a second pharmaceutical composition comprising lysine B29 (N) ε -(N α -hexadecane fatty diacid-L-lysine-N ε -oxobutanoyl)) des (B30) human insulin, preferably said second pharmaceutical composition further comprises zinc ions, wherein the content of zinc ions is 4-8 zinc ions per 6 molecules of insulin compound.
The present disclosure also provides the use of a pharmaceutical composition, product, combination, kit of any of the foregoing disclosure in the manufacture of a medicament for the treatment of diabetes, such as type 2 diabetes.
The present disclosure also provides pharmaceutical compositions, products, combinations, kits of any of the foregoing for use in treating diabetes, such as type 2 diabetes.
The present disclosure also provides a method of treating diabetes, such as type 2 diabetes, with any of the aforementioned pharmaceutical compositions, products, combinations, kits of the present disclosure comprising administering to a subject in need thereof a therapeutically effective amount of the pharmaceutical composition described above.
The disclosure also provides the use of a GLP-1 receptor agonist of formula (I) in combination with an insulin compound, such as lysine B29 (N) ε -(N α -hexadecane fatty diacid-L-lysine-N ε -oxobutanoyl)) des (B30) human insulin or a zinc complex thereof.
Drawings
FIG. 1: and (5) screening and shaking the 20D thioflavin T test result by using a zinc ion screen.
Detailed Description
Term(s)
In order that the disclosure may be more readily understood, certain technical and scientific terms are specifically defined below. Unless clearly defined otherwise herein, all other technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.
In the present disclosure, the amount of zinc ion in the composition can be expressed in terms of conventional molar concentrations (mol/L) or in terms of several zinc ions per 6 molecules of the insulin compound, illustratively 4-8 zinc ions per 6 molecules of the insulin compound, or 4-8Zn 2+ /6INS。
In the present disclosure, the term "insulin compound" refers to a peptide of native insulin or an analog or derivative thereof having insulin activity.
In the present disclosure, "native insulin" refers to naturally occurring insulin, such as human insulin, bovine insulin, or porcine insulin.
In the present disclosure, the term "insulin analogue" means a modified human insulin in which one or more amino acid residues of the insulin have been substituted by other amino acid residues and/or in which one or more amino acid residues have been deleted from the insulin and/or in which one or more amino acid residues have been added and/or inserted into the insulin.
In the present disclosure, "des (B30)" or "B (1-29)" refers to a native insulin B chain or an analog thereof lacking B30 amino acids; "A (1-21)" refers to the natural insulin A chain. For example, a21Gly, B28Asp, desB30 human insulin is an analog of human insulin in which the amino acid at position 21 in the a chain is replaced with glycine, the amino acid at position 28 in the B chain is replaced with aspartic acid, and the amino acid at position 30 in the B chain is deleted.
In the present disclosure, the term "insulin derivative" refers to a chemically modified parent insulin or an analogue thereof, wherein the modification is in the form of a linked amide, carbohydrate, alkyl, acyl, ester, pegylation, etc. For example, one example of an insulin derivative of the present disclosure is lysine B29 (N) ε -(N α -hexadecane fatty diacid-L-lysine-N ε -oxobutanoyl group) Des (B30) human insulin.
The term "receptor agonist" refers to a compound that binds to a receptor and elicits the response typical of a natural ligand. The term "GLP-1 receptor agonist" refers to compounds capable of binding to and activating the GLP-1 receptor, such as GLP-1 analogs. The term "GLP-1 analog" refers to a compound obtained by substituting a part of amino acids or extending the part of amino acids at the C-terminus to obtain a novel GLP-1 peptide comprising a human GLP-1 (7-37) peptide as a parent substance, including GLP-1 (7-36) amides and GLP-1 (7-37), and optionally chemically modifying the amino acid residue of the GLP-1 peptide by a lipophilic substituent, wherein the GLP-1 peptide has the function of human GLP-1.
The term "diabetes" includes type 1 diabetes, type 2 diabetes, gestational diabetes (during pregnancy) and other conditions that cause hyperglycemia. This term can be used for the following metabolic disorders: where the pancreas produces insufficient amounts of insulin or where the cells of the body fail to respond appropriately to insulin and thus prevent the cells from absorbing glucose, resulting in the accumulation of glucose in the blood. Type 1 diabetes, also known as insulin-dependent diabetes mellitus (IDDM) and juvenile onset diabetes, is caused by β -cell destruction, often resulting in absolute insulin deficiency. Type 2 diabetes, also known as non-insulin dependent diabetes mellitus (NIDDM) and adult onset diabetes, is associated with major insulin resistance and thus a relative insulin deficiency and/or a major insulin secretion deficiency with insulin resistance.
In the solution form of the pharmaceutical composition described in the present disclosure, the solvent is water unless otherwise specified.
"lyophilized formulation" means a pharmaceutical composition in the form of a liquid or solution or a formulation or pharmaceutical composition obtained after a vacuum freeze-drying step of a liquid or solution formulation.
By "pharmaceutical composition" or "pharmaceutical formulation" is meant a mixture containing one or more compounds described in the present disclosure, or a physiologically/pharmaceutically acceptable salt or prodrug thereof, and other chemical components, such as physiologically/pharmaceutically acceptable carriers and excipients. The purpose of the pharmaceutical composition is to maintain the stability of the polypeptide active ingredient, promote the administration to the organism, facilitate the absorption of the active ingredient and further exert the biological activity. In the present disclosure, "pharmaceutical formulation" and "pharmaceutical composition" are not mutually exclusive.
The term "about" as used in this disclosure means that the numerical value is within an acceptable error range for the particular value determined by one of ordinary skill in the art, which numerical value depends in part on how the value is measured or determined (i.e., the limits of the measurement system). For example, "about" in each practice in the art may mean within 1 or a standard deviation of more than 1. Alternatively, "about" may mean a range of up to 20%. Furthermore, particularly for biological systems or processes, the term may mean at most an order of magnitude or at most 5 times the value. Unless otherwise indicated, when a particular value appears in the application and claims, the meaning of "about" should be assumed to be within an acceptable error range for that particular value.
The pharmaceutical composition of the present disclosure can achieve a stable effect: a pharmaceutical composition wherein the protein or polypeptide substantially retains its physical and/or chemical stability and/or biological activity upon storage, preferably wherein the pharmaceutical composition substantially retains its physical and chemical stability and its biological activity upon storage. The shelf life is generally selected based on a predetermined shelf life of the pharmaceutical composition. There are a number of analytical techniques currently available for measuring protein stability, which can measure stability after storage at a selected temperature for a selected period of time.
A stable pharmaceutical polypeptide formulation is one in which no significant change is observed under the following conditions: stored at refrigeration temperatures (2-8 ℃) for at least 3 months, preferably 6 months, more preferably 1 year, and even more preferably up to 2 years. In addition, stable liquid formulations include liquid formulations that: which exhibits desirable characteristics after storage at temperatures including 25 ℃ and 40 ℃ for periods of time including 1 month, 3 months, 6 months. Typical acceptable criteria for stability are as follows: the pharmaceutical polypeptide formulation was colorless, or clear to slightly opalescent, by visual analysis. The concentration, pH and osmolality of the formulation have no more than ± 10% variation. Typically, no more than about 10%, preferably no more than about 5% truncation is observed. Typically no more than about 10%, preferably no more than about 5% aggregates are formed.
A protein or polypeptide "retains its physical stability" in a pharmaceutical formulation if it does not exhibit significant increase in aggregation, precipitation and/or denaturation as measured by visual inspection of color and/or clarity, or by UV light scattering, size Exclusion Chromatography (SEC) and Dynamic Light Scattering (DLS). Changes in protein conformation can be assessed by fluorescence spectroscopy (which determines protein tertiary structure) and by FTIR spectroscopy (which determines protein secondary structure).
A protein or polypeptide "retains its chemical stability" in a pharmaceutical formulation if it does not exhibit significant chemical alteration. Chemical stability can be assessed by detecting and quantifying the chemically altered form of the protein. Degradation processes that often alter the chemical structure of proteins include hydrolysis or truncation (assessed by methods such as size exclusion chromatography and SDS-PAGE), oxidation (assessed by methods such as peptide spectroscopy coupled to mass spectrometry or MALDI/TOF/MS), deamidation (assessed by methods such as ion exchange chromatography, capillary isoelectric focusing, peptide spectroscopy, isoaspartic acid measurement) and isomerization (assessed by measuring isoaspartic acid content, peptide spectroscopy, etc.).
A protein or polypeptide "retains its biological activity" in a pharmaceutical formulation if the biological activity of the protein or polypeptide at a given time is within a predetermined range of the biological activity exhibited at the time of preparation of the pharmaceutical formulation.
Examples
The present disclosure is further described below with reference to examples, but these examples do not limit the scope of the present disclosure. The experimental methods of the present disclosure, in which specific conditions are not specified, are generally performed according to conventional conditions, such as the antibody technical laboratory manual of cold spring harbor, molecular cloning manual; or according to the conditions recommended by the manufacturer of the raw material or the goods. Reagents of specific sources are not indicated, and conventional reagents are purchased in the market.
EXAMPLE 1 examination of preservatives and osmo-regulators
Hair brushThe insulin compound used in the examples is lysine B29 (N) ε -(N α -hexadecane fatty diacid-L-lysine-N ε -oxobutanoyl)) des (B30) human insulin (hereinafter abbreviated as INS-A), the structure of which is shown below:
Figure BDA0003642159730000121
the GLP-1 receptor agonist used has the structure shown below:
Figure BDA0003642159730000122
firstly, preparing a compound preparation according to the following process:
(1) Respectively preparing mother liquor of each auxiliary material for later use;
(2) Dissolving INS-A with se:Sup>A prescription amount in se:Sup>A proper amount of 10mM NaOH solution, adding auxiliary material mother solution with the prescription amount into INS-A main medicine solution, adding se:Sup>A proper amount of injection water, and adjusting the pH value to obtain INS-A mother solution;
(3) Adding a prescribed amount of GLP-1 receptor agonist bulk drug into 7.5mM NaOH solution, stirring, dispersing and dissolving, adjusting pH, and fixing the volume to a target volume to obtain GLP-1 receptor agonist mother liquor;
(4) Mixing INS-A mother liquor and GLP-1 receptor agonist mother liquor, uniformly stirring, adjusting pH, and fixing volume to se:Sup>A target volume of se:Sup>A preparation solution;
(5) Controlling, filtering, filling, plugging and packaging.
In order to screen preservatives and stabilizers suitable for the compound formulation, the following formulations were designed in this example: in the preparation 1, the preservative is phenol and the osmotic pressure regulator is glycerol; in the preparation 2, the preservative is phenol and m-cresol, and the osmotic pressure regulator is glycerol; in the preparation 3, the preservative is phenol, and the osmotic pressure regulator is glycerol and sodium chloride; the stability of the preparation was examined at 25 ℃ and 40 ℃ for 5 days, 10 days and 30 days (5 d, 10d and 30 d). The formulation composition is shown in table 1 below, and the results are shown in tables 2-3 below:
TABLE 1 formulation screening for preservatives and stabilizers
Figure BDA0003642159730000123
Figure BDA0003642159730000131
TABLE 2 formulation screening of preservatives and stabilizers
Figure BDA0003642159730000132
TABLE 3 formulation screening of preservatives and stabilizers
Figure BDA0003642159730000133
Figure BDA0003642159730000141
The experimental results show that the maximum single impurity, total impurity and high molecular protein of the preparation 3 (containing sodium chloride) and the preparation 2 (containing m-cresol) are increased faster compared with the preparation 1 (containing no sodium chloride and m-cresol), and the preparation 1 without the sodium chloride and the m-cresol is more excellent in stability.
EXAMPLE 2 investigation of the pH of the formulation
Because the two active ingredients have poor solubility in water and are easy to dissolve in alkaline solution, the low-concentration sodium hydroxide solution is selected as a solvent, and the pH screening is carried out on the basis of the low-concentration sodium hydroxide solution. Firstly, the pH values of the solution are selected to be 6.5, 7.0 and 8.0 for preliminary screening, and then four pH values of 7.0, 7.5, 8.0 and 8.5 are examined in detail according to the experimental results. Formulation composition at pH6.5, 7.0, 8.0 as in table 4, physicochemical results as in table 5, liquid phase results as in table 6; the formulations at pH 7.0, 7.5, 8.0, 8.5 are in table 7 and the results are shown in tables 8 and 9:
TABLE 4 pH Screen (6.5, 7.0, 8.0) formulation composition
Figure BDA0003642159730000142
TABLE 5 pH screening (6.5, 7.0, 8.0) results
Figure BDA0003642159730000143
Figure BDA0003642159730000151
TABLE 6 pH screening (6.5, 7.0, 8.0) results of investigation (continue)
Figure BDA0003642159730000152
Remarking:
[1] in the experimental process, opalescence occurs in the process of adjusting the pH to 6.0, and the solution returns to normal when the pH is adjusted to 6.5. The pH of formulation 4, formulation 5, and formulation 6 were 6.5, 7.0, and 8.0, respectively.
Test results show that the physical and chemical properties of samples with different pH values have no obvious change. The results at 40 ℃ for 30d show that with the increase of pH, the macromolecular protein, the maximum single impurity of the sample and the deamination of B3 slightly increase, the single impurity, the total impurity and the deamination of A21 obviously increase more slowly, and the INS-A content is obviously reduced when the pH is 6.5. The examination result shows that the quality of the preparation is relatively good at 7.0-8.0, so the pH values of 7.0, 7.5, 8.0 and 8.5 are selected for the next examination.
Based on the above results, solutions were screened for pH 7.0, 7.5, 8.0 and 8.5 and their stability was examined.
TABLE 7 pH Screen (7.0, 7.5, 8.0, 8.5) formulation compositions
Figure BDA0003642159730000161
TABLE 8pH screening (7.0, 7.5, 8.0, 8.5) results
Figure BDA0003642159730000162
Figure BDA0003642159730000171
TABLE 9pH screening (7.0, 7.5, 8.0, 8.5) results of investigation (continuation)
Figure BDA0003642159730000172
Test results show that the pH is between 7.0 and 8.0, and the content of the maximum single impurity, total impurity, macromolecular protein and main drug is not greatly different; the pH = 8.0-8.5, the deamination of B3, the growth rate of macromolecular protein, the maximum single impurity and total impurity are higher, and when the pH is 8.5, the GLP-1 receptor agonist content is obviously reduced; when the pH =7.3-7.8, the total impurities are minimal, the growth of each impurity and high molecular protein is moderate, and the solution is stable.
EXAMPLE 3 examination of Zinc ion concentration
The zinc ions participate in the formation of the di-and polyhexammers in the formulation. The added amount is too small, and the insulin compound molecules cannot be fully combined with the insulin compound molecules, so that the sample is easy to be unstable, and the long-acting mechanism of the medicament is influenced; the excessive addition of the zinc ions can easily cause protein aggregation, so the zinc ion concentration is screened. Under the condition of keeping INS-A, GLP-1 receptor agonist, 99.7% glycerol, phenol and pH unchanged, respectively react on 5.0Zn 2+ /6INS、6.5Zn 2+ /6INS、8.0Zn 2+ /6INS、8.5Zn 2+ /6INS(“xx Zn 2+ Xx INS "refers to the molar ratio of zinc ions to INS-se:Sup>A molecules) concentration of zinc ions, the formulation composition of which is shown in table 10, and the results are shown in table 11 below:
TABLE 10 Zinc ion screening formulation compositions
Figure BDA0003642159730000181
Note:
[1]formulation 11 was 6.5Zn 2+ 6INS; formulation 12 is 5.0Zn 2+ 6INS; formulation 13 is 8.0Zn 2+ 6INS; formulation 14 is 8.5Zn 2+ /6INS。“xx Zn 2+ The/xx INS "refers to the molar ratio of zinc ions to INS-A molecules.
[2] Formulation 14 was discarded after zinc addition, adjusting the pH to 7.5 to appear as a distinct white opalescence, turbidity, and protein aggregation associated with too high a concentration of zinc ions.
[3] The mass volume concentration of the zinc acetate is calculated by zinc acetate dihydrate.
TABLE 11 examination of Zinc ion sieves
Figure BDA0003642159730000182
The test result shows that 5.0Zn 2+ /6INS、6.5Zn 2+ /6INS、8.0Zn 2+ Under the condition of 6INS concentration zinc ions, the total impurity and the maximum single impurity of the GLP-1 receptor agonist have no obvious change. Under the standing conditions of 40 ℃ and 25 ℃, the INS-A maximum single impurity, A21 deamination and B3 deamination of each preparation and total impurity and high molecular protein have no obvious difference. The results of the thioflavin T test (figure 1) show that 6.5Zn is generated in the shaking process 2+ 6INS (preparation 11) and 8.0Zn 2+ The fibrosis degree of protein of the 6INS (preparation 13) preparation is obviously increased, which shows that the protein aggregation is easily caused by too high concentration of zinc ions, and the preparation stability is not facilitated.
The screening result of the concentration of the zinc ions shows that the zinc ions contain 5.0Zn 2+ And 6INS is more excellent, and a polymer research is developed for further verifying the result.
EXAMPLE 4 Polymer research experiment
The literature reports that the liraglutide in the similar Xultophy compound preparation can be competitively combined with insulin deguelin to form a bipeptomer with zinc ions, so that the zinc adding amount in the preparation is 8.5Zn 2+ /6INS (deglutition insulin molecule with 5 Zn) 2+ Combining; the remaining 3.5Zn 2+ For competitive binding of liraglutide) and deglu insulinAfter the preparation of the intermediate is finished, the mixture needs to be placed for 48 hours for full balance, and then the liraglutide solution is added. This example investigated whether GLP-1 receptor agonists compete for zinc ion, and the formulations used for the dimer study are shown in Table 12, and the results are shown in Table 13.
TABLE 12GLP-1 receptor agonist injection multimers study formulation composition
Figure BDA0003642159730000191
[1] The mass volume concentration of the zinc acetate is calculated by zinc acetate dihydrate.
TABLE 13 results of GLP-1 receptor agonist injection multimer study
Formulation number 15 16 17 18 19 20
Av Mw(kDa) 17.86 17.3 17.13 17.21 17.71 17.69
The research result of the polymer shows that in the GLP-1 receptor stimulant injections of different preparations, the molecular size of the GLP-1 receptor stimulant is about 17kDa, and the GLP-1 receptor stimulant exists in the form of tetramer without obvious difference. In summary, GLP-1 receptor agonists do not bind to zinc ions, so there is no need to space the INS-se:Sup>A intermediate for 48h after formulation is complete, nor is there se:Sup>A need to provide additional zinc ions.
Example 5 Long-term accelerated stability test
The recipe of table 14 was prepared as described in example 1, and left for 1, 2, 3 and 6 months at 25 ℃. + -. 2 ℃ and RH 60%. + -. 5%, and the results of accelerated condition experiments are shown in tables 15-16.
TABLE 14 formulation
Figure BDA0003642159730000201
[1] The mass volume concentration of the zinc acetate is calculated by zinc acetate dihydrate.
Watch 15
Figure BDA0003642159730000202
TABLE 16 stability of acceleration
Figure BDA0003642159730000203
The formulations in Table 14 were prepared according to the method described in example 1, and left for 3, 6, 9 and 12 months at 5 ℃. + -. 3 ℃ and RH 60%. + -. 5%, and the results of the long-term stability tests are shown in tables 17 to 18.
TABLE 17
Figure BDA0003642159730000211
TABLE 18 Long term stability
Figure BDA0003642159730000212
The long term stability test results show that the formulations of table 14 are stable.
Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, the description and examples should not be construed as limiting the scope of the disclosure. The disclosures of all patent and scientific literature cited herein are expressly incorporated by reference in their entirety.

Claims (18)

1. A pharmaceutical composition comprising an insulin compound, a GLP-1 receptor agonist and zinc ions, wherein the amount of zinc ions is between 4 and 8 zinc ions per 6 molecules of the insulin compound, said GLP-1 receptor agonist having a structure according to formula (I):
Figure FDA0003642159720000011
2. the pharmaceutical composition according to claim 1, wherein the amount of zinc ions is 5-7 zinc ions per 6 molecules of the insulin compound, preferably 5 zinc ions per 6 molecules of the insulin compound.
3. The pharmaceutical composition according to claim 1, wherein the pH of the pharmaceutical composition is 6.5-8.5, preferably 7.0-8.0, more preferably 7.3-7.8.
4. The pharmaceutical composition of claim 1, further comprising: at least one of a stabilizer, an osmotic pressure regulator, a preservative and a pH regulator;
preferably, the stabilizer is selected from one or more of zinc acetate, zinc chloride, zinc sulfate and zinc gluconate;
preferably, the tonicity modifier is selected from mannitol, sorbitol, lactose, dextrose, trehalose, sodium chloride, glycerol or any mixture thereof;
preferably, the preservative is selected from phenol, cresol, m-cresol, paraben or any mixture thereof;
preferably, the pH adjusting agent is selected from one or more of hydrochloric acid, acetic acid, sulfuric acid, phosphoric acid, sodium hydroxide, TRIS, carbonate, and phosphate.
5. The pharmaceutical composition of claim 1, comprising an insulin compound, a GLP-1 receptor agonist, glycerol, phenol, zinc acetate.
6. The pharmaceutical composition according to any one of claims 1-5, the insulin compound is selected from the group consisting of natural insulin, insulin analogs and insulin derivatives;
preferably, the natural insulin is selected from human insulin, bovine insulin and porcine insulin;
preferably, the insulin analogue is selected from Gly (a 21) Arg (B31) Arg (B32) -human insulin, lys (B3) Glu (B29) -human insulin, asp (B28) -human insulin, lys (B28) Pro (B29) -human insulin, des (B30) -human insulin;
preferably, the insulin derivative is selected from insulin deglutamide, insulin detemir and insulin derivatives having the structure shown in the following formula (II):
S-W-X-Y-Z
(II)
wherein S is human insulin with threonine at 30 th position of B chain deleted; -W-X-Y-Z is an acylated modification group of an insulin analogue;
wherein W is a compound having-OC (CH) 2 ) n A diacyl structure of CO-, wherein N is an integer between 2 and 10, which forms an amide bond with the α -amino group of the N-terminal amino acid residue of the a-chain or B-chain of the parent insulin or analog thereof or the ∈ -amino group of a lysine residue present on the B-chain with one of its acyl groups;
x is a diamino compound containing a carboxylic acid group, which is connected with one of the amino groups of the compound and one of the acyl groups in W to form an amide bond;
y is-A (CH) 2 ) m -, where m is an integer from 6 to 32, preferably an integer from 10 to 16, particularly preferably an integer from 12 to 14, A is absent or is CO-;
z is-COOH.
7. The pharmaceutical composition of claim 6, wherein the insulin derivative has the structure shown in formula (II) below:
S-W-X-Y-Z
(II)
wherein S is human insulin with threonine at 30 th position of B chain deleted; -W-X-Y-Z is an acylated modification group of an insulin analogue;
w forms an amide bond with the epsilon-amino group of a lysine residue present on the B-chain;
n is an integer between 2 and 5, preferably 2; and
x is-HN (CH) 2 ) pCH (COOH) NH-, p is an integer from 2 to 10, preferably an integer from 2 to 6, particularly preferably an integer from 2 to 4, most preferably 4.
8. The pharmaceutical composition of claim 6, wherein the insulin derivative is lysine B29 (N) ε -(N α -hexadecane fatty diacid-L-lysine-N ε -oxobutanoyl)) des (B30) human insulin.
9. The pharmaceutical composition of claim 1, comprising: lysine B29 (N) ε -(N α -hexadecane fatty diacid-L-lysine-N ε -oxobutanoyl)) des (B30) human insulin, a GLP-1 receptor agonist and zinc ions, wherein the content of zinc ions is 4-8 zinc ions per 6 molecules of the insulin compound, preferably 5-7 zinc ions per 6 molecules of the insulin compound, preferably 5 zinc ions per 6 molecules of the insulin compound.
10. The pharmaceutical composition of claim 9, further comprising glycerol and phenol.
11. A pharmaceutical composition comprising:
0.3mmol/L-1.2mmol/L lysine B29 (N) ε -(N α -hexadecane fatty diacid-L-lysine-N ε -oxobutanoyl)) des (B30) human insulin,
0.1-1mg/mLGLP-1 receptor agonist,
0.1-1.2mmol/L zinc acetate,
10-30mg/mL of glycerol, and
1-10mg/mL phenol;
preferably, the pharmaceutical composition comprises:
0.6mmol/L lysine B29 (N) ε -(N α -hexadecane fatty diacid-L-lysine-N ε -oxobutanoyl)) des (B30) human insulin,
0.24-0.4mg/mLGLP-1 receptor agonist,
0.4-0.8mmol/L zinc acetate,
10-30mg/mL of glycerol, and
1-10mg/mL phenol;
more preferably, the pharmaceutical composition comprises:
0.6mmol/L lysine B29 (N) ε -(N α -hexadecane fatty diacid-L-lysine-N ε -oxobutanoyl)) des (B30) human insulin,
0.24mg/mL to 0.4mg/mLGLP-1 receptor agonist,
0.5mmol/L of zinc acetate,
19.7mg/mL of glycerol, and
5.7mg/mL phenol.
12. The pharmaceutical composition according to claim 11, wherein the pH of the pharmaceutical composition is 7.0-8.0, preferably 7.3-7.8.
13. The pharmaceutical composition according to any one of claims 1 to 12, which is a soluble pharmaceutical composition for parenteral administration, preferably an injection, a lyophilized formulation or a solution resulting from reconstitution of a lyophilized formulation, more preferably an injection.
14. A method of preparing a pharmaceutical composition according to any one of claims 1-13, comprising:
(1) Dissolving insulin compound in 5-15mM NaOH solution, adding a stabilizer for providing zinc ions, and adjusting pH to obtain insulin compound mother liquor;
(2) Adding a GLP-1 receptor agonist into a 5-15mM NaOH solution, and adjusting the pH value to obtain a GLP-1 receptor agonist mother solution;
(3) Mixing the insulin compound mother liquor with GLP-1 receptor agonist mother liquor, and adjusting the pH;
wherein the content of zinc ions is 4-8 zinc ions per 6 insulin compound molecules, and the GLP-1 receptor agonist has a structure shown in a formula (I):
Figure FDA0003642159720000041
15. use of a pharmaceutical composition according to any one of claims 1-13 or prepared according to claim 14 for the manufacture of a medicament for the treatment of diabetes, preferably said diabetes is type 2 diabetes.
16. A pharmaceutical composition comprising an insulin compound and a GLP-1 receptor agonist, said insulin compound being lysine B29 (N) ε -(N α -hexadecane fatty diacid-L-lysine-N ε -oxobutanoyl)) des (B30) human insulin, said GLP-1 receptor agonist having a structure according to formula (I):
Figure FDA0003642159720000042
optionally, the pharmaceutical composition further comprises zinc acetate.
17. A kit comprising a first pharmaceutical composition and a second pharmaceutical composition, wherein the first pharmaceutical composition comprises a GLP-1 receptor agonist of formula (I), aThe two medicine compositions comprise lysine B29 (N) ε -(N α -hexadecane fatty diacid-L-lysine-N ε -oxobutanoyl)) des (B30) human insulin, preferably, further comprising zinc ions, wherein the content of zinc ions is 4-8 zinc ions per 6 molecules of the insulin compound.
18. Use of a GLP-1 receptor agonist of formula (I) in combination with an insulin compound, lysine B29 (N) ε -(N α -hexadecane fatty diacid-L-lysine-N ε -oxobutanoyl)) des (B30) human insulin or a zinc complex thereof.
CN202210522343.9A 2021-05-14 2022-05-13 Compound preparation of insulin compound and GLP-1 receptor agonist and medical application thereof Pending CN115337391A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115887351A (en) * 2023-01-03 2023-04-04 中科微针（北京）科技有限公司 GLP-1 receptor agonistic drug microneedle composition, microneedle prepared from same, and preparation method and application of microneedle

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115887351A (en) * 2023-01-03 2023-04-04 中科微针（北京）科技有限公司 GLP-1 receptor agonistic drug microneedle composition, microneedle prepared from same, and preparation method and application of microneedle
CN115887351B (en) * 2023-01-03 2023-05-05 中科微针（北京）科技有限公司 GLP-1 receptor agonist drug microneedle composition, microneedle prepared from same, and preparation method and application thereof

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