Fc-acylated polypeptide conjugate

Abstract

This provides compounds for managing type 2 diabetes. [Solution] A compound is provided comprising a therapeutic polypeptide Fc conjugate having activity in one or more of the following: glucose-dependent insulinotropic polypeptide (GIP), glucagon-like peptide-1 (GLP-1), and / or glucagon (GCG) receptors. These compounds have structural features that induce activity in one or more of these receptors and extend the duration of action. Methods for treating diseases and / or conditions such as obesity, chronic weight management, and type 2 diabetes are also provided.

Description

[Technical Field]

[0001] This disclosure includes, for example, glucose-dependent insulin secretion stimulating polypeptides. ent insulinotropic polypeptide (GIP), glucagon-like peptide-1 (glucagon-lik Long-term effects on peptide-1 (GLP-1), and / or glucagon (GCG) receptors Novel compounds designed to exert therapeutic effects, specifically therapeutic polypeptide F This concerns C-conjugates. These conjugates optimize receptor activity and maintain their action. By incorporating structural modifications that enhance duration, an improved pharmacological profile for therapeutic use is provided. To provide. [Background technology]

[0002] The prevalence of diabetes continues to increase, leading to the destruction of insulin secretion, insulin function, or both. This describes a chronic condition characterized by persistent hyperglycemia. (Type 2 diabetes mellitus) T2DM (us, T2DM) is the most common form, accounting for about 90% of cases. In T2DM, blood glucose levels are high. Elevated insulin levels result from a combination of impaired insulin secretion and insulin resistance. ru.

[0003] Uncontrolled diabetes in individuals affects both quality of life and overall survival. It can contribute to various health complications. Excess weight is a significant risk factor for T2DM, and T The majority of individuals with 2DM, approximately 90%, are classified as living with an overweight or obese lifestyle. Research suggests that reducing body fat may contribute to improving obesity-related health conditions. This suggests that...

[0004] The standard approach to managing T2DM is oral and injectable incretin therapy. Along with pharmacological interventions such as insulin therapy and other therapies, lifestyle changes including diet and exercise are also important. Changes will be made. Currently available injectable incretin-based therapies include dulaglutide and ce GLP-1 receptor agonists such as maglutide, as well as dual GIP and GLP-1 receptor It contains the agonist tilzepatide. A therapeutic agent containing semaglutide and tilzepatide. This involves weight loss and long-term weight management in individuals who are overweight or living with obesity. It has also been approved to support it. It has dual agonist activity at GIP and GLP-1 receptors. Further compounds are mentioned, for example, U.S. Patent Application Publication No. 20200024322 As described in issue number, as well as in GIP, GLP-1 and glucagon receptors, triple Compounds that are described as having agonist activity, for example, U.S. Patent Application Publication No. 20240 Research is being conducted on the subject described in issue 270821. Furthermore, insulin therapy The law still applies, especially to individuals with advanced disease or severe insulin deficiency. This is the foundation of diabetes management.

[0005] However, currently available injectable incretin-based therapies and insulin therapies are Generally, at least weekly administration is required. Many individuals require fewer injections. Some may prefer the option of expanding the availability of treatment to include a lower frequency of administration. Doing so has the potential to improve patient adherence, acceptance, and overall treatment success. There is.

[0006] Several approaches to enhance the duration of action of protein or peptide therapeutics This has been proposed. This approach involves conjugating the Fc region of the antibody with a therapeutic agent. (See, for example, International Publication No. 2016 / 131893) and fatty acid portion This includes lipidization (see, for example, U.S. Patent Application Publication No. 2020 / 0283492). Furthermore, A. Zaykov, et al (2024), RSC Chem. Biol. states that Lipidization and Fc conjugation affect the pharmacokinetic and pharmacodynamic profiles of insulin molecules. This section describes the effects of combining it with other products.

[0007] However, compared to currently available therapies, it reduces the frequency of administration while also increasing the duration of therapeutic effect. Innovative treatment options that offer an extension of this need are still required. This will increase patient adherence, improve treatment outcomes, and make disease management easier. We can offer a beneficial approach. [Overview of the project] [Means for solving the problem]

[0008] In the first aspect, this disclosure is based on the formula:

[0009] [ka] The present invention provides a compound of or a pharmaceutically acceptable salt thereof, wherein the formula is X TPP is a therapeutic polypeptide conjugated to a fatty acid portion, and Z is O , C(O), NH, C1~C 30 Alkyl, (OCH2CH2) m [C(O)NH-C H2CH2OCH2] m , amino acids, peptides, or combinations thereof, where m is 1 is an integer of ~30, U is C1~C5 alkyl, N, phenyl, or phenylcar bonyl, or a combination thereof, R 1 and R 2 are each independently absent, a cova lent bond, or C1~C5 alkyl, R 3 and R 4 are each independently absent or an amide, R 5 and R 6 are independen tly absent, or selected from the group consisting of C1~C5 alkyl or phenyl , which is optionally substituted with one or more of NH2, CH2NH2, and CH2CH2NH2, CH2O(CH2CH2O ) n n is an integer of 1 to 5, R 7 and R 8 are each independently absent, or

[0010]

Chemical Formula

[0011] In a second aspect, the present disclosure provides a compound of formula:

[0012]

Chemical Formula

[0013] In one embodiment, the therapeutic polypeptide acts at the GIP receptor and the GLP-1 receptor has dual agonist activity.

[0014] In a third aspect, the present disclosure provides a compound of formula:

[0015]

Chemical Structure

[0016] In one embodiment, the therapeutic polypeptide is used with GIP, GLP-1, and glucagon receptors. It possesses triple agonist activity.

[0017] In a fourth aspect, this disclosure relates to the compounds described herein, or their pharmaceutically acceptable properties. A pharmaceutical comprising a salt and at least one pharmaceutically acceptable carrier, diluent, or excipient. A composition is provided.

[0018] In a fifth embodiment, the Disclosure provides a method for treating a disease or condition, the method being effective A quantity of the compound described herein or a pharmaceutically acceptable salt thereof, as needed. This includes the process of administering the substance to the body. [Brief explanation of the drawing]

[0019] [Figure 1] The molecular structure of compound 164 (sequence number 622) is shown. [Figure 2] The molecular structure of compound 1 (sequence number 459) is shown. [Figure 3] The molecular structure of compound 32 (SEQ ID NO: 490) is shown. [Figure 4] The molecular structure of compound 47 (SEQ ID NO: 505) is shown. [Figure 5] The molecular structure of compound 127 (sequence number 585) is shown. [Figure 6] The molecular structure of compound 129 (sequence number 587) is shown. [Figure 7] The molecular structure of compound 136 (sequence number 594) is shown. [Figure 8] The molecular structure of compound 158 (sequence number 616) is shown. [Figure 9] The molecular structure of compound 168 (sequence number 626) is shown. [Figure 10] The molecular structure of compound 169 (sequence number 627) is shown. [Figure 11] The molecular structure of compound 170 (sequence number 628) is shown. [Figure 12] The molecular structure of compound 172 (sequence number 630) is shown. [Figure 13]The molecular structure of compound 173 (sequence number 631) is shown. [Figure 14] The molecular structure of compound 181 (sequence number 639) is shown. [Figure 15] The molecular structure of compound 183 (sequence number 641) is shown. [Figure 16] The molecular structure of compound 184 (sequence number 642) is shown. [Figure 17] The molecular structure of compound 185 (sequence number 643) is shown. [Figure 18] The molecular structure of compound 187 (sequence number 645) is shown. [Figure 19] The molecular structure of compound 188 (sequence number 646) is shown. [Figure 20] The molecular structure of compound 204 (sequence number 662) is shown. [Figure 21] The molecular structure of compound 479 (sequence number 1099) is shown. [Figure 22] The molecular structure of compound 480 (SEQ ID NO: 698) is shown. [Figure 23] The molecular structure of compound 481 (sequence number 699) is shown. [Figure 24] The molecular structure of compound 482 (SEQ ID NO: 700) is shown. [Figure 25] The molecular structure of compound 336 (sequence number 792) is shown. [Figure 26] The molecular structure of compound 387 (SEQ ID NO: 843) is shown. [Figure 27] The molecular structure of compound 403 (SEQ ID NO: 859) is shown. [Figure 28] The molecular structure of compound 413 (sequence number 869) is shown. [Figure 29] The molecular structure of compound 433 (sequence number 889) is shown. [Figure 30] The molecular structure of compound 435 (sequence number 891) is shown. [Figure 31] The molecular structure of compound 444 (SEQ ID NO: 900) is shown. [Figure 32] The molecular structure of compound 483 (SEQ ID NO: 1100) is shown. [Figure 33]The molecular structure of compound 484 (SEQ ID NO: 1101) is shown. [Figure 34] The molecular structure of compound 485 (SEQ ID NO: 1102) is shown. [Figure 35] The molecular structure of compound 456 (sequence number 912) is shown. [Figure 36] The molecular structure of compound 486 (sequence number 1103) is shown. [Figure 37] The molecular structure of compound 487 (SEQ ID NO: 1104) is shown. [Figure 38] The molecular structure of compound 459 (sequence number 915) is shown. [Figure 39] The molecular structure of compound 493 (SEQ ID NO: 1110) is shown. [Figure 40] The molecular structure of compound 555 (sequence number 1172) is shown. [Figure 41] The molecular structure of compound 556 (sequence number 1173) is shown. [Figure 42] The molecular structure of compound 557 (sequence number 1174) is shown. [Figure 43] The molecular structure of compound 558 (SEQ ID NO: 1175) is shown. [Figure 44] This study demonstrates that weekly administration of compound 170 (SEQ ID NO: 628) leads to a dose-dependent reduction in body weight in diet-induced obese mice. [Figure 45] This study demonstrates that weekly administration of compound 172 (SEQ ID NO: 630) leads to a dose-dependent reduction in body weight in diet-induced obese mice. [Figure 46] This study demonstrates that weekly administration of compound 387 (SEQ ID NO: 843) leads to a dose-dependent reduction in body weight in diet-induced obese mice. [Figure 47] This study demonstrates that weekly administration of compound 456 (SEQ ID NO: 912) leads to a dose-dependent reduction in body weight in diet-induced obese mice. [Modes for carrying out the invention]

[0020] This disclosure relates to a method for facilitating the delivery of therapeutic polypeptides to patients. This relates to a method and composition comprising a ptide Fc conjugate. These conjugates are A therapeutic polypeptide covalently bound to an antibody or fragment thereof, such as an Fc region, via a linker. This includes, in particular, the relationship between a therapeutic polypeptide, the disclosed linker, and an antibody or fragment thereof. The conjugation reaction occurs under relatively mild conditions, and protective measures are taken during the reaction process. Eliminate the need for steps.

[0021] Surprisingly, acylated therapeutic polypeptides are linked to the linkers disclosed herein. By using this method to bind an antibody or a fragment thereof, for example, to the Fc region, therapeutic poly The duration of action of the peptide depends on either acylation or conjugation to the Fc region. The linker provides significantly greater improvement and / or extension than would be expected when used alone. - And the binding of therapeutic polypeptides to the Fc region, the hydrodynamic size of the entire portion, FcR To increase n-bonding and / or reuse, and to prevent the degradation of therapeutic polypeptides. One or more of these factors may increase the duration of action of therapeutic polypeptides. Conjugate between the linker and antibody or antibody fragment, e.g., Fc region, as disclosed in the specification. Because the reaction conditions for the reaction are relatively mild, therapeutic polypeptides can be retained during the reaction. It does not need to be modified to protect it. Acylation of therapeutic polypeptides is performed on serum albumin By promoting binding to the peptide, one or more of these mechanisms can prolong the action of the therapeutic polypeptide. The interval can be further increased. The duration of action of therapeutic polypeptides, in certain embodiments, By including structural features that improve the proteolytic stability of therapeutic polypeptides, This could increase even further.

[0022] Therapeutic polypeptide Fc conjugate This disclosure describes several different embodiments of compounds that include features resulting in increased duration of action. provide.

[0023] In certain embodiments, this disclosure relates to formula I:

[0024] [ka] The present invention provides a compound of or a pharmaceutically acceptable salt thereof, wherein, Therapeutic Polypeptide X TPP It is conjugated in the fatty acid portion, and Z is a linker - and U is C1-C5 alkyl, N, phenyl, or phenylcarbonyl, These are combinations, R 1 and R 2 They either exist independently or are covalently bonded. Or C1-C5 alkyl, R 3 and R 4 It is either independent, nonexistent, or an amide. And R 5 and R 6 These are either not present independently, or C1-C5 alkyl or flu Selected from the group consisting of enyls, which include NH2, CH2NH2, and CH2CH2NH 2. CH2O(CH2CH2O) n One or more of these can be arbitrarily selected to replace n, where n is 1 to 5 It is an integer of R 7 and R 8 It is either independent, does not exist, or

[0025] [ka] Selected from the group consisting of, In the formula, ( * ) is R 9 and R 10 It is a connection point, (** ) is R 5 or R 6 Connection point And R 9 and R 10 Each of these is an antibody or a fragment thereof.

[0026] In certain embodiments, Z is O, C(O), NH, C1~C 30 Alkyl, (OCH2 CH2) m [C(O)NH-CH2CH2OCH2] m , (2-[2-(2-amino- Ethoxy)-Ethoxy]-Acetyl) m , amino acids, peptides, or combinations thereof It must contain one or more of the following, and m is an integer between 1 and 30.

[0027] In a particular embodiment, R 1 and R 2 They are the same. In a particular embodiment, R 3 and R 4 They are the same. In a particular embodiment, R 5 and R 6 They are the same. In a particular embodiment, R 7 and R 8 They are the same. In a particular embodiment, R 9 and R 10 This is the Fc region. In a specific embodiment, R 9 and R 10 They are the same.

[0028] In certain embodiments, the therapeutic polypeptide is GIP, GLP-1 and / or glucago It possesses agonist activity in one or more of the receptors.

[0029] In some embodiments, the present disclosure provides compounds of formula I, where R 7 and R 8 teeth,

[0030] [ka] That is the case.

[0031] In some embodiments, R 5 and R 6 These are each C2 alkyl groups substituted with NH2. In some embodiments, R 3 and R 4 These are all amides. In this embodiment, R 5 and R 6 Each of these is a C2 alkyl group. In some embodiments, U is either N or phenyl.

[0032] In some embodiments, this disclosure relates to formula IA:

[0033] [ka] The present invention provides compounds or pharmaceutically acceptable salts thereof.

[0034] In some embodiments, the present disclosure provides compounds of formula I, where R 7 and R 8 teeth,

[0035] [ka] That is the case.

[0036] In some embodiments, R 5 and R 6 These are (OCH2CH2) m Replaced by It is phenyl, and m is 3. In some embodiments, R 3 and R 4 These are A It is mid. In some embodiments, R 1 and R2 These are each C2 alkyl groups. In some embodiments, U is N or phenyl.

[0037] In some embodiments, this disclosure uses formula IB:

[0038] [ka] The present invention provides compounds or pharmaceutically acceptable salts thereof.

[0039] In some embodiments, the present disclosure provides compounds of formula I, where R 7 and R 8 teeth,

[0040] [ka] That is the case.

[0041] In some embodiments, R 5 and R 6 These are (OCH2CH2) m Replaced by It is phenyl, and m is 3. In some embodiments, R 3 and R 4 These are A It is mid. In some embodiments, R 1 and R 2 These are each C2 alkyl groups. In some embodiments, U is N or phenyl.

[0042] In some embodiments, the present disclosure relates to formula IC:

[0043] [ka] The present invention provides compounds or pharmaceutically acceptable salts thereof.

[0044] In some embodiments of the compound of formula I, R 7 and R 8 None of them exist. How many? In that embodiment, R 5 and R 6 Each of these is a C1 alkyl group. In some embodiments is, R 3 and R 4 Each of these is an amide. In some embodiments, R 1 and R 2 haso Each is a C2 alkyl group. In some embodiments, U is nitrogen or phenyl.

[0045] In some embodiments, this disclosure refers to formula ID:

[0046] [ka] The present invention provides compounds or pharmaceutically acceptable salts thereof.

[0047] In some embodiments, this disclosure relates to formula IE:

[0048] [ka] The present invention provides compounds or pharmaceutically acceptable salts thereof.

[0049] In certain embodiments, this disclosure relates to Formula II:

[0050] [ka] The present invention provides a compound of or a pharmaceutically acceptable salt thereof, wherein, X TPP is a therapeutic polypeptide, and L is conjugated to the therapeutic polypeptide. It is a linker, R 9 and R 10 , and R9 and R 10 Each is either an antibody or its antibody It is a fragment. In a particular embodiment, R 9 and R 10 These are each Fc regions.

[0051] In some embodiments, therapeutic polypeptides are used with GIP receptors and GLP-1 receptors. It has dual agonist activity. In some embodiments, the therapeutic polypeptide is It possesses triagonist activity at GIP, GLP-1, and glucagon receptors.

[0052] In certain embodiments, this disclosure relates to Formula III:

[0053] [ka] The present invention provides a compound of or a pharmaceutically acceptable salt thereof, wherein, Fc1 and Fc2 are either non-existent or are Fc regions, respectively, and L is in the linker. Yes, Z is a functional group of a small molecule, amino acid polymer, or combination thereof, and X is It is a therapeutic polypeptide, and the therapeutic polypeptide is conjugated with fatty acid Y at an optional rate. It is being treated as such.

[0054] In certain embodiments, this disclosure relates to formula IV:

[0055] [ka] The present invention provides a compound of or a pharmaceutically acceptable salt thereof, wherein, R 1 and R 2 It does not exist, or independently,

[0056] [ka] Selected from the group consisting of, in the formula, ( * ) is a connection point to the sulfur atom in the cysteine ​​residue of Fc1 or Fc2, ( ** ) is R 3 or R 4 It is a connection point to R 3 and R 4 These are independently C1-C5 alkyl groups. This is an arbitrary selection of one or more of NH2, CH2NH2, and CH2CH2NH2. Replaced by a choice, R 5 and R 6 These are independently covalent or C1-C5 alkyl groups. U is a tertiary amine or a 1,3,5-substituted phenyl, and Z is O, NH, C1-C 30 Alkyl, (OCH2CH2) m , amino acid polymers, or combinations thereof m is an integer between 1 and 30, X is a therapeutic polypeptide, and Y is either non-existent or non-existent. Alternatively, it is a fatty acid, and Fc1 and Fc2 are independently Fc regions.

[0057] In some embodiments, this disclosure relates to formula IV-A:

[0058] [ka] The present invention provides a compound of or a pharmaceutically acceptable salt thereof, wherein, R 3 and R 4 These are independently C1-C5 alkyl groups, which are NH2, CH2NH 2, and one or more of CH2CH2NH2 are optionally substituted, R 5 and R 6 teeth, Independently, the bonds are covalent or C1-C5 alkyl, and U is a tertiary amine or 1,3 It is a ,5-substituted phenyl, where Z is O, NH, C1~C 30 Alkyl, (OCH2CH2 ) m , amino acid polymers, or combinations thereof, where m is an integer from 1 to 30. X is a therapeutic polypeptide, Y is absent or a fatty acid, and Fc1 and F c2 is an independent Fc region.

[0059] In another embodiment, this disclosure is,

[0060] [ka] The present invention provides a compound of formula IV-A containing or a pharmaceutically acceptable salt thereof, wherein, R3 and R4 are independently C1-C5 alkyl groups, which are NH2 and CH2NH2. 2, and one or more of CH2CH2NH2 are optionally substituted, R 5 and R 6 teeth, Independently, they are covalent or C1-C5 alkyl, and Z is O, NH, C1-C 30 Alkyl, (OCH2CH2) m , amino acid polymers, or combinations thereof, m is an integer between 1 and 30, X is a therapeutic polypeptide, and Y is either nonexistent or These are fatty acids, and Fc1 and Fc2 are independently Fc regions.

[0061] In yet another embodiment, the Disclosure

[0062] [ka] The present invention provides a compound of formula IV-A containing or a pharmaceutically acceptable salt thereof, wherein, R3 and R4 are independently C1-C5 alkyl groups, which are NH2 and CH2NH2. 2, and one or more of CH2CH2NH2 are optionally substituted, and Z is O, NH, C1-C30 alkyl groups, (OCH2CH2) m , amino acid polymers, or combinations thereof It is a combination of values, where m is an integer from 1 to 30, X is a therapeutic polypeptide, and Y is present. It is either absent or a fatty acid, and Fc1 and Fc2 are independently Fc regions.

[0063] In some embodiments, this disclosure relates to formula IV-B:

[0064] [ka] The present invention provides a compound of or a pharmaceutically acceptable salt thereof, wherein, R 3 and R 4 These are independently C1-C5 alkyl groups, which are NH2, CH2NH 2, and one or more of CH2CH2NH2 are optionally substituted, R 5 and R 6 teeth, Independently, the bonds are covalent or C1-C5 alkyl, and U is a tertiary amine or 1,3 It is a ,5-substituted phenyl, where Z is O, NH, C1~C 30 Alkyl, (OCH2CH2 ) m , amino acid polymers, or combinations thereof, where m is an integer from 1 to 30. X is a therapeutic polypeptide, Y is absent or a fatty acid, and Fc1 and F c2 is an independent Fc region.

[0065] In another embodiment, this disclosure is,

[0066] [ka] The present invention provides a compound of formula IV-B containing or a pharmaceutically acceptable salt thereof, wherein, R 3 and R 4 These are independently C1-C5 alkyl groups, which are NH2, CH2NH 2, and one or more of CH2CH2NH2 are optionally substituted, R 5 and R 6 teeth, Independently, they are covalent or C1-C5 alkyl, and Z is O, NH, C1-C 30 Alkyl, (OCH2CH2) m , amino acid polymers, or combinations thereof, m is an integer between 1 and 30, X is a therapeutic polypeptide, and Y is either nonexistent or These are fatty acids, and Fc1 and Fc2 are independently Fc regions.

[0067] In yet another embodiment, the Disclosure

[0068] [ka] The present invention provides a compound of formula IV-B containing or a pharmaceutically acceptable salt thereof, wherein, R 3 and R 4 These are independently C1-C5 alkyl groups, which are NH2, CH2NH 2, and one or more of CH2CH2NH2 are optionally substituted, and Z is O, NH, C1~C 30 Alkyl, (OCH2CH2) m , amino acid polymers, or combinations thereof It is a combination of values, where m is an integer from 1 to 30, X is a therapeutic polypeptide, and Y is present. It is either absent or a fatty acid, and Fc1 and Fc2 are independently Fc regions.

[0069] In some embodiments, this disclosure relates to formulas IV-C:

[0070] [ka] The present invention provides a compound of or a pharmaceutically acceptable salt thereof, wherein, R 3 and R 4 These are independently C1-C5 alkyl groups, which are NH2, CH2NH 2, and one or more of CH2CH2NH2 are optionally substituted, R 5 and R 6 teeth, Independently, the bonds are covalent or C1-C5 alkyl, and U is a tertiary amine or 1,3 It is a ,5-substituted phenyl, where Z is O, NH, C1~C 30 Alkyl, (OCH2CH2 ) m , amino acid polymers, or combinations thereof, where m is an integer from 1 to 30. X is a therapeutic polypeptide, Y is absent or a fatty acid, and Fc1 and F c2 is an independent Fc region.

[0071] In another embodiment, this disclosure is,

[0072] [ka] The present invention provides a compound of formula IV-C containing or a pharmaceutically acceptable salt thereof, wherein, R 3 and R 4 These are independently C1-C5 alkyl groups, which are NH2, CH2NH 2, and one or more of CH2CH2NH2 are optionally substituted, R 5 and R 6 teeth, Independently, they are covalent or C1-C5 alkyl, and Z is O, NH, C1-C 30 Alkyl, (OCH2CH2) m , amino acid polymers, or combinations thereof, m is an integer between 1 and 30, X is a therapeutic polypeptide, and Y is either nonexistent or These are fatty acids, and Fc1 and Fc2 are independently Fc regions.

[0073] In yet another embodiment, the Disclosure

[0074] [ka] The present invention provides a compound of formula IV-C containing or a pharmaceutically acceptable salt thereof, wherein, R 3 and R 4 These are independently C1-C5 alkyl groups, which are NH2, CH2NH 2, and one or more of CH2CH2NH2 are optionally substituted, and Z is O, NH, C1~C 30 Alkyl, (OCH2CH2) m , amino acid polymers, or combinations thereof It is a combination of values, where m is an integer from 1 to 30, X is a therapeutic polypeptide, and Y is present. It is either absent or a fatty acid, and Fc1 and Fc2 are independently Fc regions.

[0075] In some embodiments, this disclosure relates to formula IV-D:

[0076] [ka] The present invention provides a compound of or a pharmaceutically acceptable salt thereof, wherein, R 3 and R 4 These are independently C1-C5 alkyl groups, which are NH2, CH2NH 2, and one or more of CH2CH2NH2 are optionally substituted, R 5 and R 6 teeth, Independently, the bonds are covalent or C1-C5 alkyl, and U is a tertiary amine or 1,3 It is a ,5-substituted phenyl, where Z is O, NH, C1~C 30 Alkyl, (OCH2CH2 ) m , amino acid polymers, or combinations thereof, where m is an integer from 1 to 30. X is a therapeutic polypeptide, Y is absent or a fatty acid, and Fc1 and F c2 is an independent Fc region.

[0077] In another embodiment, this disclosure is,

[0078] [ka] The present invention provides a compound of formula IV-D containing or a pharmaceutically acceptable salt thereof, wherein, R 3 and R 4 These are independently C1-C5 alkyl groups, which are NH2, CH2NH 2, and one or more of CH2CH2NH2 are optionally substituted, R 5 and R 6 teeth, Independently, they are covalent or C1-C5 alkyl, and Z is O, NH, C1-C 30 Alkyl, (OCH2CH2) m , amino acid polymers, or combinations thereof, m is an integer between 1 and 30, X is a therapeutic polypeptide, and Y is either nonexistent or These are fatty acids, and Fc1 and Fc2 are independently Fc regions.

[0079] In yet another embodiment, the Disclosure

[0080] [ka] The present invention provides a compound of formula IV-D containing or a pharmaceutically acceptable salt thereof, wherein, R3 and R 4 These are independently C1-C5 alkyl groups, which are NH2, CH2NH 2, and one or more of CH2CH2NH2 are optionally substituted, and Z is O, NH, C1~C 30 Alkyl, (OCH2CH2) m , amino acid polymers, or combinations thereof It is a combination of values, where m is an integer from 1 to 30, X is a therapeutic polypeptide, and Y is present. It is either absent or a fatty acid, and Fc1 and Fc2 are independently Fc regions.

[0081] Polypeptide conjugate for intermediate therapy This disclosure provides intermediate compounds comprising therapeutic polypeptides, the therapeutic polypeptides being The signal is conjugated, followed by an antibody or an antibody fragment, for example, conjugated to the Fc region. Gate. Non-restrictive examples of such intermediate compounds are provided below.

[0082] Maleimide intermediate compound In some embodiments, this disclosure is,

[0083] [ka] The present invention provides a maleimide intermediate compound of formula IV-A containing or a pharmaceutically acceptable salt thereof, and formula Medium, R 3 and R 4 These are independently C1-C5 alkyl groups, which are NH2, CH2N H2 and one or more of CH2CH2NH2 are optionally substituted, R 5 and R 6 teeth Independently, the bonds are covalent or C1-C5 alkyl, and U is a tertiary amine or 1. It is a 3,5-substituted phenyl compound, where Z is O, NH, C1~C 30 Alkyl, (OCH2CH 2) m , amino acid polymers, or combinations thereof, where m is an integer from 1 to 30. X is a therapeutic polypeptide, and Y is either absent or a fatty acid.

[0084] In another embodiment, this disclosure is,

[0085] [ka] The present invention provides a maleimide intermediate compound of formula IV-A containing or a pharmaceutically acceptable salt thereof, and formula Medium, R 3 and R 4 These are independently C1-C5 alkyl groups, which are NH2, CH2N H2 and one or more of CH2CH2NH2 are optionally substituted, R 5 and R 6 teeth Independently, the bonds are covalent or C1-C5 alkyl, and Z is O, NH, C1-C3 0 alkyl, (OCH2CH2) m , amino acid polymers, or combinations thereof m is an integer from 1 to 30, X is a therapeutic polypeptide, and Y is either nonexistent or It is a fatty acid.

[0086] In yet another embodiment, the Disclosure

[0087] [ka] The present invention provides a maleimide intermediate compound of formula IV-A containing or a pharmaceutically acceptable salt thereof, and formula Medium, R 3 and R 4 These are independently C1-C5 alkyl groups, which are NH2, CH2N H2 and one or more of CH2CH2NH2 are optionally substituted, and Z is O, NH , C1~C 30 Alkyl, (OCH2CH2) m , amino acid polymers, or combinations thereof It is a combination where m is an integer from 1 to 30, X is a therapeutic polypeptide, and Y is present. It is either absent or contains fatty acids.

[0088] [ka]

[0089] Furthermore, this specification discloses compounds having the above structure or pharmaceutically acceptable salts thereof, wherein, R 3 and R 4 These are independently C1-C5 alkyl groups, which are NH2, CH2NH2 , and one or more of CH2CH2NH2 are optionally substituted, R 5 and R 6 German It is either covalently bonded or C1-C5 alkyl, and U is a tertiary amine or 1,3. It is a 5-substituted phenyl compound, where Z is O, NH, C1~C 30 Alkyl, (OCH2CH2) m , amino acid polymers, or combinations thereof, where m is an integer from 1 to 30, X is a therapeutic polypeptide, Y is absent or is a fatty acid, and D1 and D2 are Independently, amides, amines, imines, sulfonamides, thioureas, ureas, thioethers, Or a combination of these.

[0090] [ka]

[0091] Furthermore, this specification discloses compounds having the above structure or pharmaceutically acceptable salts thereof, wherein, R 3 and R 4These are independently C1-C5 alkyl groups, which are NH2, CH2NH2 , and one or more of CH2CH2NH2 are optionally substituted, R 5 and R 6 German It is either covalently bonded or C1-C5 alkyl, and U is a tertiary amine or 1,3. It is a 5-substituted phenyl compound, where Z is O, NH, C1~C 30 Alkyl, (OCH2CH2) m , amino acid polymers, or combinations thereof, where m is an integer from 1 to 30, X Y is a therapeutic polypeptide, and Y is either absent or a fatty acid.

[0092] [ka]

[0093] Furthermore, this specification discloses compounds having the above structure or pharmaceutically acceptable salts thereof, wherein, R 3 and R 4 These are independently C1-C5 alkyl groups, which are NH2, CH2NH2 , and one or more of CH2CH2NH2 are optionally substituted, R 5 and R 6 German It is either a covalent bond or a C1-C5 alkyl group, and Z is O, NH, C1-C 30 a Lukil, (OCH2CH2) m , amino acid polymers, or combinations thereof, m X is an integer between 1 and 30, X is a therapeutic polypeptide, and Y is either absent or a fat. It is an acid.

[0094] [ka]

[0095] Furthermore, this specification discloses compounds having the above structure or pharmaceutically acceptable salts thereof, wherein, R 3 and R 4 These are independently C1-C5 alkyl groups, which are NH2, CH2NH2 , and one or more of CH2CH2NH2 are optionally substituted, and Z is O, NH, C 1~C 30 Alkyl, (OCH2CH2) m , amino acid polymers, or combinations thereof The formula is as follows: m is an integer between 1 and 30, X is a therapeutic polypeptide, and Y does not exist. Or, it is a fatty acid.

[0096] Tetrazole intermediate compounds In some embodiments, this disclosure is,

[0097] [ka] The present invention provides a tetrazole intermediate compound of formula IV-B containing or a pharmaceutically acceptable salt thereof, In the formula, R 3 and R 4 These are independently C1-C5 alkyl groups, which are NH2, CH2 NH2 and one or more CH2CH2NH2 are optionally substituted, R 5 and R 6 The atoms are independently covalent or C1-C5 alkyl, and U is a tertiary amine or 1 It is a ,3,5-substituted phenyl, and Z is O, NH, C1~C 30 Alkyl, (OCH2C H2) m It is an amino acid polymer, or a combination thereof, where m is an integer from 1 to 30. X is a therapeutic polypeptide, and Y is either absent or a fatty acid.

[0098] In another embodiment, this disclosure is,

[0099] [ka] The present invention provides a tetrazole intermediate compound of formula IV-B containing or a pharmaceutically acceptable salt thereof, In the formula, R 3 and R 4 These are independently C1-C5 alkyl groups, which are NH2, CH2 NH2 and one or more CH2CH2NH2 are optionally substituted, R 5 and R 6 The elements are independently covalent or C1-C5 alkyl, and Z is O, NH, C1-C 30 Alkyl, (OCH2CH2) m , amino acid polymers, or combinations thereof m is an integer between 1 and 30, X is a therapeutic polypeptide, and Y is either nonexistent or It is a fatty acid.

[0100] In yet another embodiment, the Disclosure

[0101] [ka] The present invention provides a tetrazole intermediate compound of formula IV-B containing or a pharmaceutically acceptable salt thereof, In the formula, R 3 and R 4 These are independently C1-C5 alkyl groups, which are NH2, CH2 NH2 and CH2CH2NH2 are optionally substituted, and Z is O, N H, C1~C 30 Alkyl, (OCH2CH2) m , amino acid polymers, or combinations thereof It is a combination, m is an integer from 1 to 30, X is a therapeutic polypeptide, and Y is present. Either it is not, or it is a fatty acid.

[0102] [ka]

[0103] Furthermore, this specification discloses compounds having the above structure or pharmaceutically acceptable salts thereof, wherein, R 3 and R 4 These are independently C1-C5 alkyl groups, which are NH2, CH2NH2 , and one or more of CH2CH2NH2 are optionally substituted, R 5 and R 6 German It is either covalently bonded or C1-C5 alkyl, and U is a tertiary amine or 1,3. It is a 5-substituted phenyl compound, where Z is O, NH, C1~C 30 Alkyl, (OCH2CH2) m , amino acid polymers, or combinations thereof, where m is an integer from 1 to 30, X is a therapeutic polypeptide, Y is absent or is a fatty acid, and D1 and D2 are Independently, amides, amines, imines, sulfonamides, thioureas, ureas, thioethers, Or a combination of these.

[0104] [ka]

[0105] Furthermore, this specification discloses compounds having the above structure or pharmaceutically acceptable salts thereof, wherein, R 3 and R 4 These are independently C1-C5 alkyl groups, which are NH2, CH2NH2 , and one or more of CH2CH2NH2 are optionally substituted, R 5 and R 6 German It is either covalently bonded or C1-C5 alkyl, and U is a tertiary amine or 1,3. It is a 5-substituted phenyl compound, where Z is O, NH, C1~C 30 Alkyl, (OCH2CH2) m , amino acid polymers, or combinations thereof, where m is an integer from 1 to 30, X Y is a therapeutic polypeptide, and Y is either absent or a fatty acid.

[0106] [ka]

[0107] Furthermore, this specification discloses compounds having the above structure or pharmaceutically acceptable salts thereof, wherein, R 3 and R 4 These are independently C1-C5 alkyl groups, which are NH2, CH2NH2 , and one or more of CH2CH2NH2 are optionally substituted, R 5 and R 6 German It is either covalently bonded or C1-C5 alkyl, and Z is O, NH, C1-C 30 a Lukil, (OCH2CH2) m , amino acid polymers, or combinations thereof, m X is an integer between 1 and 30, X is a therapeutic polypeptide, and Y is either absent or a fat. It is an acid.

[0108] [ka]

[0109] Furthermore, this specification discloses compounds having the above structure or pharmaceutically acceptable salts thereof, wherein, R 3 and R 4 These are independently C1-C5 alkyl groups, which are NH2, CH2NH2 , and one or more of CH2CH2NH2 are optionally substituted, and Z is O, NH, C 1~C 30 Alkyl, (OCH2CH2) m , amino acid polymers, or combinations thereof The formula is as follows: m is an integer between 1 and 30, X is a therapeutic polypeptide, and Y does not exist. Or, it is a fatty acid.

[0110] Oxydisol intermediate compounds In some embodiments, this disclosure is,

[0111] [ka] The present invention provides a tetrazole intermediate compound of formula IV-C containing or a pharmaceutically acceptable salt thereof, In the formula, R 3 and R 4 These are independently C1-C5 alkyl groups, which are NH2, CH2 NH2 and one or more CH2CH2NH2 are optionally substituted, R 5 and R 6 The atoms are independently covalent or C1-C5 alkyl, and U is a tertiary amine or 1 It is a ,3,5-substituted phenyl, and Z is O, NH, C1~C 30 Alkyl, (OCH2C H2) m It is an amino acid polymer, or a combination thereof, where m is an integer from 1 to 30. X is a therapeutic polypeptide, and Y is either absent or a fatty acid.

[0112] In another embodiment, this disclosure is,

[0113] [ka] The present invention provides a tetrazole intermediate compound of formula IV-C containing or a pharmaceutically acceptable salt thereof, In the formula, R 3 and R4 These are independently C1-C5 alkyl groups, which are NH2, CH2 NH2 and one or more CH2CH2NH2 are optionally substituted, R 5 and R 6 The elements are independently covalent or C1-C5 alkyl, and Z is O, NH, C1-C 30 Alkyl, (OCH2CH2) m , amino acid polymers, or combinations thereof m is an integer between 1 and 30, X is a therapeutic polypeptide, and Y is either nonexistent or It is a fatty acid.

[0114] In yet another embodiment, the Disclosure

[0115] [ka] The present invention provides a tetrazole intermediate compound of formula IV-C containing or a pharmaceutically acceptable salt thereof, In the formula, R 3 and R 4 These are independently C1-C5 alkyl groups, which are NH2, CH2 NH2 and CH2CH2NH2 are optionally substituted, and Z is O, N H, C1~C 30 Alkyl, (OCH2CH2) m , amino acid polymers, or combinations thereof It is a combination, m is an integer from 1 to 30, X is a therapeutic polypeptide, and Y is present. Either it is not, or it is a fatty acid.

[0116] [ka]

[0117] Furthermore, this specification discloses compounds having the above structure or pharmaceutically acceptable salts thereof, wherein, R3 and R 4 These are independently C1-C5 alkyl groups, which are NH2, CH2NH2 , and one or more of CH2CH2NH2 are optionally substituted, R 5 and R 6 German It is either covalently bonded or C1-C5 alkyl, and U is a tertiary amine or 1,3. It is a 5-substituted phenyl compound, where Z is O, NH, C1~C 30 Alkyl, (OCH2CH2) m , amino acid polymers, or combinations thereof, where m is an integer from 1 to 30, X is a therapeutic polypeptide, Y is absent or is a fatty acid, and D1 and D2 are Independently, amides, amines, imines, sulfonamides, thioureas, ureas, thioethers, Or a combination of these.

[0118] [ka]

[0119] Furthermore, this specification discloses compounds having the above structure or pharmaceutically acceptable salts thereof, wherein, R 3 and R 4 These are independently C1-C5 alkyl groups, which are NH2, CH2NH2 , and one or more of CH2CH2NH2 are optionally substituted, R 5 and R 6 German It is either covalently bonded or C1-C5 alkyl, and U is a tertiary amine or 1,3. It is a 5-substituted phenyl compound, where Z is O, NH, C1~C 30 Alkyl, (OCH2CH2) m , amino acid polymers, or combinations thereof, where m is an integer from 1 to 30, X Y is a therapeutic polypeptide, and Y is either absent or a fatty acid.

[0120] [ka]

[0121] Furthermore, this specification discloses compounds having the above structure or pharmaceutically acceptable salts thereof, wherein, R 3 and R 4 These are independently C1-C5 alkyl groups, which are NH2, CH2NH2 , and one or more of CH2CH2NH2 are optionally substituted, R 5 and R 6 German It is either covalently bonded or C1-C5 alkyl, and Z is O, NH, C1-C 30 a Lukil, (OCH2CH2) m , amino acid polymers, or combinations thereof, m X is an integer between 1 and 30, X is a therapeutic polypeptide, and Y is either absent or a fat. It is an acid.

[0122] [ka]

[0123] Furthermore, this specification discloses compounds having the above structure or pharmaceutically acceptable salts thereof, wherein, R 3 and R 4 These are independently C1-C5 alkyl groups, which are NH2, CH2NH2 , and one or more of CH2CH2NH2 are optionally substituted, and Z is O, NH, C 1~C 30 Alkyl, (OCH2CH2) m , amino acid polymers, or combinations thereof The formula is as follows: m is an integer between 1 and 30, X is a therapeutic polypeptide, and Y does not exist. Or, it is a fatty acid.

[0124] In some embodiments described herein, the therapeutic polypeptide is conjugated to Z. Z is composed of O, C(O), NH, C1-C30 alkyl, (OCH2CH2) m [C(O)NH-CH2CH2OCH2] m , (2-[2-(2-amino-ethoxy) (-ethoxy]-acetyl) m , amino acids, peptides, or one of a combination thereof It includes one or more, and m is an integer from 1 to 30. In some embodiments, Z is one of the following Contains peptides that include one or more sequences.

[0125] [Table 1]

[0126] Linker In some embodiments, formula II represents a therapeutic polypeptide as an antibody or a fragment thereof, for example. , including a linker L for connecting to the Fc region. In certain embodiments, L in formula II is

[0127] [ka] Including, in the formula, ( *** ) includes a connection point to a therapeutic polypeptide, ( * ) is R 9 or R 10 Includes a connection point to [the destination].

[0128] In some other embodiments, L in formula II is

[0129] [ka] Including, in the formula, (*** ) includes a connection point to a therapeutic polypeptide, ( * ) is R 9 or R 10 Includes a connection point to [the destination].

[0130] In some embodiments, the present disclosure connects Z to another part via a nucleophilic conjugate addition reaction. It contains linker L. In some embodiments, the portion is in the Fc region of the antibody or fragment thereof. It is one or more cysteine ​​amino acids in the Fc region. In some embodiments, the portion is the Fc region. This is a sulfur atom from a cysteine ​​residue in the region. In some embodiments, Z is as follows: As shown in some of the reactions illustrated and described, amides, amines, and imines , sulfonamide, thiourea, urea, thioether, thiol, cysteine, or these This occurs via nucleophilic conjugate addition reactions to other nucleophiles, such as combinations of these agents.

[0131] In some embodiments, the linker is configured in the above formulas IV-A, IV-B, and IV-C. As shown, it may include a central trivalent linking unit U. In some embodiments, one of U The portion can be bonded to Z. In some embodiments, the other two portions of U are maleimide. A functional group, for example, an electrophile, is used to add two Zs in a nucleophilic conjugate addition reaction. It can selectively bind to the nucleophile.

[0132] Therefore, the linker of the present invention has two or more maleimide tubes before the nucleophilic conjugate addition reaction. The functional group, preferably two maleimide functional groups, is provided. Alternatively, the linker of the present invention is aromatic Before the nucleophilic substitution reaction, two or more methylsulfonyltetrazole functional groups, or preferably, It contains two methylsulfonyltetrazole functional groups. Alternatively, the linker of the present invention is Before the aromatic nucleophilic substitution reaction, two or more methylsulfonyloxadiazole functional groups, or Preferably, it contains two methylsulfonyloxadiazole functional groups. Alternatively, the present invention The linker is formed by adding two or more bromoacetyl functional groups, or preferably two, before the nucleophilic substitution reaction. It contains two bromoacetyl functional groups.

[0133] Suitable trivalent linkage units are trisubstituted C5-C8 aryls, with 1-3 heteroatoms in the ring system. Trisubstituted C5-C8 heteroaryl, trisubstituted C5-C8 cycloalkyl, 1 in the ring system Trisubstituted C5-C8 heterocycles with ~3 heteroatoms, tertiary amines, tertiary phosphines It may contain any atom or molecule that is a trisubstituted, such as a combination of these. In other words, U is a 1,3,5-trisubstituted phenyl or tertiary amine.

[0134] In some embodiments, the structures bonded to U are shown below.

[0135] [ka]

[0136] In these structures above, R3 can be a C1-C5 alkyl, which is -NH2. -CH2NH2 and -CH2CH2NH2 are optionally substituted, R 5 may or may not be a C1-C5 alkyl group. R4 is a C1-C5 alkyl group. Possible, -NH 2、 -CH2NH 2、 And one or more of the following: -CH2CH2NH2 Selective substitution occurs, and R6 may be a C1-C5 alkyl group or it may not exist.

[0137] R3 and / or R4 are preferably C2 alkyl, -NH2, -CH2NH 2, or one or more of -CH2CH2NH2 are optionally substituted. R5 and / or R6 is preferably a C2 alkyl when U is a tertiary amine, and U is 1 It does not exist in the case of ,3,5-trisubstituted phenyl.

[0138] If both of the above structures are bonded to U, R3 and R4 may be the same, or Independently, these may be C1-C5 alkyl groups, such as -NH2, -CH2NH2, and One or more of the following structures are optionally substituted: When coupled to U, R5 and R6 may be the same, or independently, C1 to C5 It may be alkyl, or it may not be present. If R5 and / or R6 are not present In addition, the amide nitrogen in the structure is directly connected to U, for example, when U is a 1,3,5-trisubstituted phenyl. They are connected.

[0139] The third arm of the trivalent linked unit bonded to U may include Z. Z is O, NH, C1~ C 30 Alkyl, (OCH2CH2) m , amino acid polymers, or combinations thereof Including m, where m is an integer from 1 to 30, X is a therapeutic polypeptide, and Y is either nonexistent or non-existent. , or fatty acids, and Fc1 and Fc2 are independently Fc regions. Z is U or ka It can be directly connected to a carbonyl functional group. For example, the carbon atoms of the carbonyl functional group are U and Z. They can be directly joined.

[0140] Alternatively, each arm bonded to the U and tetrazole functional groups can be represented by the following structure: can.

[0141] [ka]

[0142] Alternatively, each arm bonded to the U and oxadiazole functional groups may be represented by the following formula: It is possible.

[0143] [ka]

[0144] In one embodiment, the first arm of the two arms represented by the above structure is R3 is phenyl-(OCH2CH2) m OCH2 is possible, and R5 is C1~C5 It may or may not be a kill. In another embodiment, 2 represented by the above structure R4, the second of the two arms, is phenyl-(OCH2CH2) m OCH It can be 2, and R6 can be a C1-C5 alkyl group, or it may not exist.

[0145] R3 and / or R4 may preferably be phenyl-(OCH2CH2)2OCH2. R5 and / or R6 are preferably C2 alkyl when U is a tertiary amine. This is possible, but it does not exist when U is a 1,3,5-trisubstituted phenyl compound.

[0146] When both arms are coupled to U, R3 and R4 may be the same or independent. It may also be phenyl-(OCH2CH2)2OCH2. Both arms are connected to U. If they are combined, R5 and R6 may be the same, or independently, C1~C5 are all-in-one. It may be a ru, or it may not exist. If R5 and / or R6 do not exist, above In the structure described above, the amide nitrogen is directly attached to U, for example, when U is a 1,3,5-trisubstituted phenyl compound. Connecting.

[0147] The third of the three arms connected to U includes Z. Z is H, OH, NH2, NH, therapeutic drugs, C1~C 30 Alkyl, (OCH2CH2) m , or these combinations It may include combinations, and m is an integer from 1 to 30. Z can be directly connected to U, or Z can be attached to a carbonyl functional group. The carbon atoms of the carbonyl functional group are U and It can be directly coupled to Z.

[0148] Alternatively, each arm bonded to the U and acetyl functional groups can be represented by the following structure: ru.

[0149] [ka]

[0150] In one embodiment, the first arm of the two arms represented by the above structure is R3 can be a C1-C5 alkyl group, and R5 can be a C1-C5 alkyl group, or It does not exist. In another embodiment, of the two arms represented by the above structure, Arm 2, R4, can be C1-C5 alkyl, and R6 can be C1-C5 alkyl. It may or may not exist.

[0151] R3 and / or R4 may preferably be C2 alkyl. R5 and / or R6 are Preferably, if U is a tertiary amine, it can be a C2 alkyl group, and U is 1,3,5- This does not exist in the case of trisubstituted phenyl compounds.

[0152] When both arms are coupled to U, R3 and R4 may be the same or independent. And it may be a C1-C5 alkyl group. If both arms are bonded to U, R5 And R6 may be the same, or independently may be C1-C5 alkyl groups, It does not have to exist. If R5 and / or R6 do not exist, then expression VB or expression VC The amide nitrogen is directly attached to U, for example, when U is a 1,3,5-trisubstituted phenyl compound. ru.

[0153] The third of the three arms connected to U includes Z. Z is H, OH, NH2, NH, therapeutic drugs, C1~C 30 Alkyl, (OCH2CH2) m , or these combinations It may include combinations, and m is an integer from 1 to 30. Z can be directly connected to U, and Z can be attached to a carbonyl functional group. The carbon atom of the carbonyl functional group is given by formula As shown in II-IV, it can be directly coupled to U and Z.

[0154] The reaction conditions for conjugation are relatively mild, therefore the payload, i.e. The therapeutic agent does not need to be modified to protect during the reaction. Disclosed in the art. Other linkers that are used include therapeutic polypeptides, such as insulin, in the recombinant extension. Modification is required. The conjugated compounds disclosed herein have a longer duration of action. It is long-lasting, does not require modification of the therapeutic polypeptide, and is effective.

[0155] [ka]

[0156] In some embodiments disclosed herein, compounds of the above structure and their pharmaceutically acceptable properties are described. The salt is acceptable, and in the formula, R3 and R4 are independently selected from C1-C5 alkyl groups. R5 and R6 are independently C1-C5 alkyl groups or are absent, and U is third It is a quaternary amine or a 1,3,5-substituted phenyl, where Z is O, NH, C1-C 30 Alkyl (OCH2CH2) m X contains amino acid polymers, or combinations thereof, and is used for treatment. It contains polypeptides, Y contains fatty acids, m is an integer from 1 to 30, and Fc1 and Fc Each of the 2s includes an Fc region.

[0157] [ka]

[0158] In some embodiments disclosed herein, compounds of the above structure or pharmaceutically acceptable compounds thereof The salt is acceptable, and in the formula, R3 and R4 are independently selected from C1-C5 alkyl groups. R5 and R6 are independently C1-C5 alkyl groups, and Z is O, NH, C1-C3 0 alkyl, (OCH2CH2) m , amino acid polymers, or combinations thereof X contains therapeutic polypeptides, Y contains fatty acids, m is an integer from 1 to 30, and Fc 1 and Fc2 each contain an Fc region.

[0159] [ka]

[0160] In some embodiments disclosed herein, compounds of the above structure or pharmaceutically acceptable compounds thereof The salt is acceptable, and in the formula, R3 and R4 are independently selected from C1-C5 alkyl groups. Z is O, NH, C1~C 30 Alkyl, (OCH2CH2) m , amino acid polymer, Or a combination thereof, where X contains a therapeutic polypeptide, Y contains a fatty acid, m is an integer between 1 and 30, and Fc1 and Fc2 each contain the Fc region.

[0161] [ka]

[0162] In some embodiments disclosed herein, compounds of the above structure or pharmaceutically acceptable compounds thereof The salt is acceptable, and in the formula, R3 and R4 are independently selected C1-C5 alkyl groups. Yes, R5 and R6 are independently C1-C5 alkyl or absent, and U is It is a tertiary amine or a 1,3,5-substituted phenyl, where Z is O, NH, C1-C 30 Al Kill, (OCH2CH2) m X includes amino acid polymers, or combinations thereof. Contains therapeutic polypeptides, Y contains fatty acids, m is an integer from 1 to 30, and D1 and D 2 independently includes amides, amines, imines, sulfonamides, thioureas, ureas, and thio- Tel, or combinations thereof.

[0163] [ka]

[0164] In some embodiments disclosed herein, compounds of the above structure or pharmaceutically acceptable compounds thereof The salt is acceptable, and in the formula, R3 and R4 are independently selected from C1-C5 alkyl groups. R5 and R6 are independently C1-C5 alkyl groups or are absent, and U is third It is a quaternary amine or a 1,3,5-substituted phenyl, where Z is O, NH, C1-C 30 Alkyl (OCH2CH2) m X contains amino acid polymers, or combinations thereof, and is used for treatment. It contains polypeptides, Y contains fatty acids, and m is an integer between 1 and 30.

[0165] [ka]

[0166] In some embodiments disclosed herein, compounds of the above structure or pharmaceutically acceptable compounds thereof The salt is acceptable, and in the formula, R3 and R4 are independently selected C1-C5 alkyl groups. Yes, R5 and R6 are C1-C5 alkyl, and Z is O, NH, C1-C 30 Al Kill, (OCH2CH2) m X includes amino acid polymers, or combinations thereof. It contains therapeutic polypeptides, Y contains fatty acids, and m is an integer between 1 and 30.

[0167] [ka]

[0168] In some embodiments disclosed herein, compounds of the above structure or pharmaceutically acceptable compounds thereof The salt is acceptable, and in the formula, R3 and R4 are independently selected from C1-C5 alkyl groups. Z is O, NH, C1~C 30 Alkyl, (OCH2CH2) m , amino acid polymer, Or a combination thereof, where X contains a therapeutic polypeptide, Y contains a fatty acid, m The integer is between 1 and 30.

[0169] In some embodiments, the compounds of the present disclosure are antibodies or as shown in Formula II. In the Fc region of that fragment, one or more cysteine ​​amino acids or other parts, or this As shown in the structure disclosed in the specification, amides, amines, imines, sulfonamides, Thiourea, urea, thioether, thiol, cysteine, or combinations thereof, etc. Another suitable nucleophile contains a linker L that connects Z via a nucleophilic conjugate addition reaction.

[0170] In some embodiments, the linker is as shown in the structures disclosed herein. It may contain a central trivalent linking unit U. One of the linker parts may bond to Z. Yes, it is possible. The other two parts of the linker contain a maleimide functional group, an electrophile, and a nucleophilic conjugate. In additive reactions, Z can be used to selectively attach to two nucleophiles.

[0171] Nucleophilic conjugate addition reaction with maleimide functional group As disclosed herein, linker L is coupled to Z and linked payload This may result in Z further including therapeutic agents X, for example, linked therapeutic agents. Several implementations In this state, linker L contains two maleimide functional groups. Maleimide functional groups are nucleophilically conjugated. To bind Z to different parts of the antibody or its fragment, such as the Fc region, via an addition reaction. It can be used for this purpose.

[0172] In nucleophilic conjugate addition reactions, maleimide is an electrophile and reacts with the nucleophile to form an electrophilic carbon atom. A new covalent bond is formed between the atom and the nucleophilic atom, which is the maleimide bond as shown below. This could lead to ring opening of the active group.

[0173] [ka]

[0174] Various nucleophiles can be used to react with electrophilic maleimide functional groups. Examples of nucleophiles include moieties containing one or more nitrogen atoms and / or sulfur atoms in the functional group. Therefore, this nitrogen atom and / or sulfur atom can function as an electron-rich nucleophile. Nucleophile Appropriate examples include amides, amines, imines, sulfonamides, thiols, thioureas, Examples include parts containing one or more functional groups of urea or a combination thereof, and these This is shown in the following structure.

[0175] [ka]

[0176] In particular, the amino acid cysteine ​​is found in many peptides, antibodies, and proteins, as shown below. These are found in various substances, and contain thiol functional groups, which are used in peptides, antibodies, and tannins. Between the protein and the linker, the linked payload, and / or the linked therapeutic agent It can be used as a nucleophile for forming thioethers.

[0177] [ka]

[0178] The conditions for nucleophilic conjugate addition reactions are well known to those skilled in the art. The conditions used for a particular reaction are selected. The nucleophilic and electrophilic compounds may vary depending on the selected nucleophile and electrophile. As disclosed herein, the nucleophilic compounds Compounds and / or compounds containing a linked payload attached to another part via an addition reaction There are conjugated compounds.

[0179] Methylsulfonyl-tetrazole functional group or methylsulfonyl-oxadiazole functional group Nucleophilic aromatic substitution (SNAr) reactions with groups As disclosed herein, linker L is coupled to Z and linked payload This may result in Z further including therapeutic agents X, for example, linked therapeutic agents. Several implementations In this state, linker L consists of two methylsulfonyl-tetrazole functional groups or two methyl Contains a sulfonyl-oxadiazole functional group. Also contains a methylsulfonyl-tetrazole functional group. The methylsulfonyl-oxadiazole functional group is nucleophilic aromatic substitution, which can lead to the formation of antibodies or This can be used to combine Z with different parts of the fragment, such as the Fc region.

[0180] In nucleophilic aromatic substitution reactions, methylsulfonyl or oxadiazole of tetrazoles. The methylsulfonyl is substituted with a nucleophile, and as shown below, it is replaced by a nucleophile atom and an electrophilic carbon atom. It acts as a leaving group, forming a new covalent bond with its parent.

[0181] [ka]

[0182] Various nucleophiles have an electrophilic methylsulfonyltetrazole functional group or methylsulfonyl It can be used to react with oxadiazole functional groups. Suitable nucleophiles include functional groups. Examples include a portion containing one or more nitrogen atoms and / or sulfur atoms in the base, and this nitrogen atom and / or sulfur atoms can function as electron-rich nucleophiles. A suitable example of a nucleophile is: The following are examples of thiol-containing portions.

[0183] [ka]

[0184] In particular, the amino acid cysteine ​​is found in many peptides, antibodies, and proteins, as shown below. These are found in various substances, and contain thiol functional groups, which are used in peptides, antibodies, and tannins. Between the protein and the linker, the linked payload, and / or the linked therapeutic agent It can be used as a nucleophile for forming thioethers.

[0185] [ka]

[0186] The conditions for aromatic nucleophilic substitution reactions are well known to those skilled in the art. The conditions used for specific reactions are... The flavor may vary depending on the selected nucleophile and electrophile. As disclosed herein, the aroma Compounds containing linked payloads attached to another part via a nucleophilic substitution reaction and There are bi / or conjugated compounds.

[0187] Nucleophilic substitution with bromoacetyl functional group (S N 2) Reaction As disclosed herein, linker L is coupled to Z and linked payload This may result in, or if Z contains a therapeutic agent, it may result in a linked therapeutic agent. The linkers disclosed in the details are two or more, or preferably two, as shown in formula VD. It contains two bromoacetyl functional groups. The bromoacetyl functional group undergoes a nucleophilic substitution reaction, which is equivalent to F It can be used to combine Z into different regions, such as the c region.

[0188] In nucleophilic substitution reactions, the bromide of bromoacetyl is substituted with a nucleophile, as shown below. It acts as a leaving group that forms a new covalent bond between a nucleophilic atom and an electrophilic carbon atom. ru.

[0189] [ka]

[0190] Various nucleophiles can be used to react with electrophilic bromoacetyl functional groups. Suitable nucleophiles include those containing one or more nitrogen and / or sulfur atoms in their functional group. A portion of this is the nitrogen atom and / or sulfur atom, which function as an electron-rich nucleophile. To obtain. Suitable examples of nucleophiles include amides, amines, imines, sulfonamides, and thiols. Examples include a portion containing one or more functional groups of thiourea, urea, or a combination thereof. These are shown in the following structure.

[0191] [ka]

[0192] In particular, the amino acid cysteine ​​is found in many peptides, antibodies, and proteins, as shown below. These are found in various substances, and contain thiol functional groups, which are used in peptides, antibodies, and tannins. Between the protein and the linker, the linked payload, and / or the linked therapeutic agent It can be used as a nucleophile for forming thioethers.

[0193] [ka]

[0194] The conditions for nucleophilic substitution reactions are well known to those skilled in the art. The conditions are selected for use in specific reactions. The reaction may vary depending on the nucleophile and electrophile used. Disclosed herein are nucleophilic substitution reactions. Compounds and / or conjugated compounds containing a linked payload attached to another part via a linkage That is the case.

[0195] Duration of action The compounds described herein have the characteristic of extending the duration of action of therapeutic polypeptides. These features include the conjugation of antibodies or fragments thereof, for example, to the Fc region. Albumin-binding moieties, e.g., acylation of therapeutic polypeptides with fatty acids, and / or Incorporation of structural features into the therapeutic polypeptide itself to enhance the protein degradation stability. This includes. In certain embodiments, compounds containing each or all of these features are extended It has an extended duration of action and requires less frequent administration compared to currently available therapies. To enable, for example, one or more of GIP, GLP-1, and / or glucagon receptors. In the context of therapeutic polypeptides having binding and activity, as described herein Certain compounds can be used in treatment methods involving infrequent medication, such as once a month. It has a long duration of effect.

[0196] Furthermore, this specification also discloses a method for increasing the duration of action of a therapeutic agent. This may include a step of binding a therapeutic agent to a trivalent linker to obtain a linker therapeutic agent.

[0197] This method involves a nucleophilic addition or substitution reaction to convert a linked therapeutic agent into an antibody or its cleavage. This may further include a step of conjugating at least a portion of the piece, for example, the Fc region. These reactions involve a linked therapeutic agent and two nucleophiles present in the Fc region (sulfur atom and / This occurs with (or anions, etc.), leading to the formation of conjugated compounds.

[0198] The resulting conjugated compound comprises a therapeutic molecule covalently bonded to the disclosed linker, which is, Next, it is conjugated to an Fc fragment. Conjugation enhances the action of the unbound therapeutic agent. It extends the duration of action beyond its normal duration.

[0199] In certain embodiments, the linker disclosed herein is a linker whose reaction conditions are such that other linkers It is noteworthy because it is milder compared to other nucleophilic addition or substitution reactions disclosed using this method. It offers advantages that should be met. For example, the disclosed linker can break down antibodies or their anastomoses with minimal heating. It can be combined with one Fc region. Specifically, the disclosed linker is 30°C At a temperature not exceeding, or possibly 25°C, the linked therapeutic agent and the reduced Fc region By mixing the region for 2 hours or less, the Fc region of the antibody or its fragment is conjugated. It is possible.

[0200] Linker preparation Preparation 1 4-(bis(2-((tert-butoxycarbonyl)amino)ethyl)amino)-4 -Oxobutanoic acid

[0201] [ka] A solution of succinic anhydride (0.990 g, 9.88 mmol) in DCM (40 mL) , 10-oxa-2,5,8-triazadodecanoic acid, 11,11 in DCM (40 mL) -Dimethyl-9-oxo-,1,1-dimethylethyl ester (3.0g, 9.88mm) The mixture was added to the solution of (ol) under N2 conditions at ambient temperature for 30 minutes. The reactants were then heated at ambient temperature for 48 minutes. The mixture was stirred for a certain amount of time, diluted with H2O (50 mL), and extracted with DCM (3 × 40 mL). The organic layer was then separated. The mixture was combined and washed with saturated NH4Cl aqueous solution, NaHCO3 aqueous solution, and brine. The layers were dried with MgSO4, filtered, and concentrated in a vacuum. The material was then analyzed using DCM to extract siliceous gel. The material was adsorbed onto silica, then concentrated under reduced pressure to obtain a free-flowing residue. The residue was then collected from normal-phase silica. Purify by gel chromatography, then DCM (5 min), followed by DCM / MeOH (9:1). Elution was performed to obtain the title compound (2.84 g, 70%) as a white powder. ES / MS (m / z):402(M-).

[0202] Preparation 2 1-(4-(5-(methylsulfonyl)-1H-tetrazole-1-yl)phenoxy )-12-(2-(2-(2-(2-(4-(5-(methylsulfonyl)-1H-tetra Zol-1-yl)phenoxy)ethoxy)ethoxy)acetamido)ethyl)-8,1 3-Dioxo-3,6-Dioxa-9,12-Diazahexadecane-16-Eucic Acid

[0203] [ka] 4-(bis(2-((tert-butoxycarbonyl)amino)ethyl)amino)-4 - Dissolve oxobutanoic acid (0.47g, 1.05 mmol) in DCM (3mL), then... TFA (2 mL) was then added. The reaction mixture was stirred at ambient temperature for 3 hours, and then concentrated in a vacuum. Toluene (10 mL) was added to the residue, and the mixture was concentrated in a vacuum. The process was repeated once more. The residue was dried under vacuum for 2 hours, and then added to DMF (5 mL). It dissolved. DIEA (0.41g, 3.15 mmol) was added, followed by 2,5-dioxypropyl alcohol. Sopyrroridine-1-yl2-(2-(2-(4-(5-(methylsulfonyl)-1H-te Trazol-1-yl phenoxyethoxyethoxyacetate (1.07g, 2 A 0.10 mmol (4 mL) solution of DMF was added. The reaction mixture was stirred at ambient temperature for 1 hour. Next, it was diluted with DCM (100 mL), washed with brine, and then with H2O. The organic layer The solution was dried with Na2SO4, filtered, and concentrated in vacuum. The residue was then processed using normal-phase silica gel chromatography. Purification was performed by tography. The column was run with DCM for 5 minutes, followed by 0% to 20% methanol. Elution was performed over 20 minutes using 2L / DCM to obtain the title compound (0.86g, 82%) as a white solid. The result was obtained as follows: ES / MS (m / z): 940 (M+H).

[0204] Preparation 3 (S)-2-((tert-butoxycarbonyl)amino)-4-(2,5-dioxo -2,5-dihydro-1H-pyrrole-1-yl)butanoic acid

[0205] [ka]

[0206] Add triethylamine (1 mL, 7.17 mmol) to 1,4-dioxane (10 mL) , (2S)-4-amino-2-(tert-butyrate) in THF (5 mL) and water (5 mL) Added to a mixture of xycarbonylaminobutanoic acid (500 mg, 2.29 mmol) Stir until a homogeneous mixture is obtained, then add methyl 2,5-dioxo-2,5-dihydroxy Add dro-1H-pyrrole-1-carboxylate (370 mg, 2.31 mmol). Mixed at ambient temperature for 1 hour. Dilute with 50 mL of water and add 5N HCl. The pH was adjusted to approximately 6. The aqueous solution was mixed with HCl (30 mL) and chloroform / Extraction was performed with sopropanol (3 x 30 mL). The organic phases were combined and dried on sodium sulfate. The mixture was filtered and concentrated under reduced pressure. The resulting residue was then mixed with 0-30% MeOH in DCM. Purified by eluting silica gel chromatography, the title compound (580 mg, 8 A score of 1% was obtained. ES / MS m / z: 199 (M-tBu).

[0207] Preparation 4 2,5-Dioxopyrrolidine-1-yl(S)-2-((tert-butoxycarbonyl (Lu)amino)-4-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl) ) Butanoate

[0208] [ka] Dicyclohexylcarbodiimide (320 mg, 1.53 mmol) is used in THF (6 m (S)-2-((tert-butoxycarbonyl)amino)-4-(2,5-di) Oxo-2,5-dihydro-1H-pyrrole-1-yl)butanoic acid (400 mg, 1.2 Dissolve 7 mmol) and N-hydroxysuccinimide (180 mg, 1.53 mmol) It was added to the liquid. The mixture was stirred at ambient temperature for 3 hours. The solid was removed by filtration, and the filtrate was then reduced under reduced pressure. The compound was concentrated to obtain the title compound (605 mg, 84%), which was then used without purification. . ES / MS m / z:296(M-tBu).

[0209] Preparation 5 (S)-2-((tert-butoxycarbonyl)amino)-3-(2,5-dioxo -2,5-dihydro-1H-pyrrole-1-yl)propanoic acid

[0210] [ka] (2S)-3-amino-2-(tert-butoxycarbonylamino)propanoic acid (5 (0.00g, 24.5 mmol) in a 1M sodium bicarbonate aqueous solution (130 mL, 130 mL) Mixed with (mol). Stirred at ambient temperature for 10 minutes until a clear solution was formed. Cool it in an ice bath and methyl 2,5-dioxo-2,5-dihydro-1H-pyrrole-1 - Add carboxylate (4.00g, 25.0 mmol) in three separate additions over 15 minutes. The mixture was stirred continuously at 0°C for 3 hours. 100 mL of toluene was added and stirred under cooling conditions. Next, concentrated HCl was added to adjust the pH to 1. The layers were separated and measured in a DCM (4 × 50 mL) The aqueous solution was extracted. The organic layers were combined, washed with saturated NaCl solution, and dried with sodium sulfate. The material was dried. The solvent was removed under reduced pressure. Silica gel eluted in 0-40% MeOH in DCM. The title compound (5.70 g, 66%) was purified by chromatography. / MS m / z:185(M-tBu).

[0211] Preparation 6 2,5-Dioxopyrrolidine-1-yl(S)-2-((tert-butoxycarbonyl (Lu)amino)-3-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl) )propanoate

[0212] [ka] Dicyclohexylcarbodiimide (484 mg, 2.32 mmol) is used in THF (5 m (S)-2-((tert-butoxycarbonyl)amino)-3-(2,5-di) Oxo-2,5-dihydro-1H-pyrrole-1-yl)propanoic acid (550 mg, 1. (54 mmol) and N-hydroxysuccinimide (272 mg, 2.31 mmol) It was added to the mixture. The mixture was stirred at ambient temperature until a precipitate appeared. The solid was removed by filtration. The sample was washed with DCM. The filtrate was concentrated under reduced pressure to obtain the title compound (1.18 g, 85%). ES / MS m / z: 282 (M+H-Boc).

[0213] Fc area In certain embodiments, a nucleophile that can react with two or more functional groups on a linker is This is the Fc region, also known as the fragment crystallizable region. The Fc region consists of CH2 and CH3 The lower constant domain consists of a complete or partial hinge together with the domain or a portion thereof. This Fc fragment has an effector function by completely binding to the Fc receptor. This may include mutations that either obtain or restrict or disable Fc receptor binding. In particular, IgA, F is a specific antibody class, such as IgD, IgE, IgG, IgM, or combinations thereof. The c region consists of two or more disulfide bridges that form within the N-terminal portion of the molecule or hinge region. It may contain cysteine ​​residues.

[0214] In some embodiments, the Fc region of the antibody is linked to a linker via a nucleophilic conjugate addition reaction. Modifications may occur before binding. Such modifications can lead to unintended reactivity between the antibody and the linker. It can be designed to minimize noise and ensure that the desired nucleophilic coupling reaction proceeds efficiently. I'll do that.

[0215] In certain embodiments, the Fc region is used for smoothing specific segments such as hinge regions or This could be a modified version of the antibody's Fc domain, which may have changes including removal.

[0216] The appropriate disulfide bridges within the Fc region are reduced to the linker described herein. Two nucleophiles that can undergo nucleophilic conjugate addition reactions with maleimide functional groups. This can result in a thiol group.

[0217] Alternatively, reduced disulfide crosslinks in the appropriate Fc region are as described herein. The linker has a methylsulfonyltetrazole functional group or methylsulfonyl oxadiazole functional group. This may provide a thiol nucleophile that can participate in aromatic nucleophilic addition reactions with thiol functional groups. .

[0218] Exemplary Fc region In this specification, the term "Fc region" includes at least a portion of the steady region. This term is used to define the C-terminal region of immunoglobulin heavy chains. Includes the c region and the variant Fc region.

[0219] As used herein, "Variant Fc region", "Fc region variant", or The term "Fc region mutation" refers to a mutation in the parent Fc region caused by at least one amino acid substitution (or This refers to the amino acid sequence of the Fc region that is different from the sequence of that fragment.

[0220] As used herein, “amino acid substitution” means at least one amino acid in a given amino acid sequence. This refers to replacing one existing amino acid residue with another different "substitute" amino acid residue.

[0221] Furthermore, in this specification, substitution is defined as the amino acid in the parent Fc, followed by the position where the substitution occurs. The name is derived from the number, followed by the amino acid that substituted for the amino acid in the parent Fc region at the same position. For example, the human IgG1 Fc region variant P247I is a variant of the parent human IgG1 F This shows that the proline residue at position 247 of the c region is substituted with an isoleucine residue. ).

[0222] In one embodiment, the human IgG heavy chain Fc region is derived from Cys229, and the heavy chain carboxyl It extends to the terminal. However, the C-terminal lysine (Lys447) of the Fc region is present even if It is not necessary. Unless otherwise specified herein, in the Fc region or steady region The numbering of amino acid residues is based on Kabat et al., Sequences of P roteins of Immunological Interest,5th Ed .Public Health Service,National Institute EU number listed in es of Health, Bethesda, MD, 1991 It follows a metric index (also known as the EU index).

[0223] In certain embodiments, the human IgG4 Fc amino acid sequence (hIgG4) is recrosslinked. It is used for judging.

[0224] [Table 2]

[0225] The Fc region of sequence number 935 is an additional N composed of alanine and glycine (AG). A blunt hinge having a terminal dipeptide, starting at position 229 (EU index numbering) It contains natural IgG4. To minimize the effector function, Fc is bold and As indicated by the underline, the CH2 domain contains F234A and L235A (EU index It contains two mutations (numbered as 'X'). Unless otherwise specified, the following are listed in Table 3. The reference to "Fc" in the compound nomenclature described in the specification refers to Sequence ID No. 935. To point.

[0226] IgG4 Fc region containing mutations in Q274K, Q355R, and E419Q (SEQ ID NO: 936) Biophysics when more neutral peptides are used in conjugation To enhance its properties, it can be used to increase the overall pI of Fc.

[0227] [Table 3]

[0228] Unless otherwise specified, the compound nomenclature described in Table 3 below and as used herein The reference to "Fc KRQ" in this context refers to sequence number 936.

[0229] To mitigate incorrect processing of the N-terminus in the CHO expression system, high titer and 1 Several specific signaling peptides provide minimal N-terminal clipping of alanine residues. I identified Do.

[0230] [Table 4]

[0231] Instead of altering the signal peptide, replace the N-terminal alanine with cysteine. C), arginine (R), isoleucine (I), leucine (L), tyrosine (Y), and We searched for six alternative residues containing rheonine (T), but all of them prevented clipping. Alternatively, any of the 20 amino acids excluding alanine, serine, and glycine The N-terminal residue can be replaced using any of the following. For example, the N-terminal residue of isoleucine. Terminal Fc(Fc IG KRQ) and cysteine ​​N-terminal KRQ Fc(Fc N-Cys KRQ) is one example, and these are F234A, L, and so on, as shown in the following sequences. Includes the 235A, Q274K, Q355R, and E419Q mutations.

[0232] [Table 5]

[0233] Another example is the cysteine ​​N-terminal Fc (Fc N-Cys), which is... The sequence below includes the F234A and L235A mutations.

[0234] [Table 6]

[0235] Unless otherwise specified, the nomenclature of compounds shown in Table 3 below is "Fc IG K The reference to "RQ" refers to Sequence ID No. 941, and in the nomenclature of compounds shown in Table 3 below... The reference to "Fc N-Cys KRQ" refers to sequence number 942, which is shown in Table 3 below. The reference to "Fc N-Cys" in the nomenclature of the compound refers to Sequence ID No. 1178. .

[0236] In addition to the IgG4 subclass, IgG1 Fc also has an effector knockout mutation F. IgG1 with 234A, L235A, and P329A (EU index numbering) We used [this method] to search.

[0237] [Table 7]

[0238] In some embodiments, wild-type IgG1 Fc having full effector functionality is used. It can be used as a substitute for the effects knockout version.

[0239] [Table 8]

[0240] As an alternative to the above recrosslinking approach, the manipulated cysteine ​​was initiated at position 216, IgG4 Fc 358, 398, and IgG4 Fc 358, 398, which have a complete hinge containing the S228P mutation. It was introduced at position 415 to enhance hinge stability. The Fc fragment was used in two operated systems. It contains a symmetric molecule having half or two peptides per Fc conjugate. To drip.

[0241] [Table 9]

[0242] Fc having a single peptide conjugated via manipulated cysteine To achieve this, the Fc has a complete hinge starting at position 216 and lacks the eCys part, C Generated using mutations F405L and R409K in the H3 domain. Fc exchange approach. Using this method, a half-Fc lacking the eCys site is paired with a half-Fc containing the desired eCys site. Then, conjugation was performed.

[0243] [Table 10]

[0244] In certain embodiments, the compounds of the present disclosure are SEQ ID NOs: 935-936, 941-948, At least 90% sequence identity with any of 1178-1181, or at least 95% sequence identity, or at least 97% sequence identity, or at least 98% sequence identity It includes an Fc region that is identical or has at least 99% sequence identity.

[0245] General methodology and conditions for FC addition Scheme 1

[0246] [ka] Scheme 1, process A starts with RAPP AM-Rink Amide resin, C Using the Fmoc / t-Bu strategy with the lassic Symphony automated peptide synthesizer H2N-PEG produced by solid-phase peptide synthesis 24 - Through peptide synthesis, This shows how to obtain compound (2). Step B involves PEGylated peptide (2) and bis-Fmoc. The trivalent core (U is either a 1,3,5-trisubstituted phenyl or a tertiary amine (Preparation 1)) This shows that compound (3) is provided by the reaction with ( ). In step C, Fmoc The group is removed, and the resulting primary amine is converted into the NHS ester MalDap (commercial product), MalD Compound (4) is produced by reacting ab (preparation 15) or β-MalDap (preparation 17) with ab. Obtained. In step D, the peptide was cleaved from the resin. The bismaleimide functional group is in the Fc region. The two reducing thiol functional groups react with a nucleophilic conjugate addition reaction to form conjugate (5). I got it.

[0247] Scheme 2

[0248] [ka] Scheme 2, Step A is the production of H2N-PEG by solid-phase peptide synthesis. 24 -pe This shows that compound (1) is obtained by the synthesis of butyl. Step B is the PEGylated peptide (2 1) Coupling of bis-(methylsulfonyl)tetrazole analog (Preparation 12) This demonstrates the provision of compound (2). In step C, the peptide is cleaved from the resin. The bis-(methylsulfonyl)tetrazole functional group was added via a nucleophilic conjugate addition reaction. The two reducing thiol functional groups in the c region were reacted to obtain conjugate (3).

[0249] Scheme 3

[0250] [ka] Scheme 3, Step A is the production of H2N-PEG by solid-phase peptide synthesis. 24 -pe This shows that compound (1) is obtained by the synthesis of butyl. Step B is the PEGylated peptide (2 ) and bis-Fmoc protected trivalent core (U is a 1,3,5-trisubstituted phenyl or tertiary amine (conditioning) This demonstrates that compound (2) is provided by coupling with any of the products (1). In step C, the Fmoc group is removed, and the resulting primary amine is converted to 2,5-dioxopyrrolic acid. Compound (3) is provided by reacting it with din-1-yl-2-bromoacetate. 3) The functional group is combined with two reducing thiol functional groups in the Fc region in a nucleophilic conjugate addition reaction. The reaction was carried out to obtain conjugate (4).

[0251] Therapeutic polypeptides The term "therapeutic polypeptide" refers to the use in the treatment and / or prevention of a disease or condition. This refers to any amino acid polymer having suitable activity for use. Conjugate structure as used herein It is used to provide an extended duration of action for various therapeutic polypeptides. Such drugs may include, but are not limited to, the following, or their variants. Examples include: GIP, GLP-1, GLP-2, GIP, and oxytomodulin (OXM). ), amylin, apelin, urocortin (UCN), ANP, BNP, CNP, parathyroid gland Hormones (parathyroid hormone, PTH), MC4 agonists, somatostatin, GD F15, Peptide YY (PYY), Pancreatic polypeptide, ACTH, VIP, Oxytocin ghrelin, gastrin, insulin, human growth hormone (hG) H), fibroblast growth factor 21 (FGF21), inter Leukin-2 (interleukin 2, IL2), IL10, IL15, IL22, IL7, Peptide Statins, calcitonin, erythropoietin (EPO), thrombopoietin and receptors Agonist, angiotensin, vasopressin, thymosin β-4, guanylate cyclase -C agonist, epidermal growth factor (EGF), α-1-anti Trypsin, corticotropin-releasing hormone, CRH, GnRH, Thymosin α1, Follicle-stimulating hormone , FSH), thyroid stimulating hormone (TSH), Thyroid-releasing hormone (TRH), melanocortin-stimulating hormone (MEL) Anacortin stimulating hormone (MSH), apo-CII mimetic peptide, factor IX, Factor VIII, Factor X, Granulocyte Colony-Stimulating Factor, Interferon-α, β-Amygdala Lloyd's inhibitors, CD40 ligand, CD137 agonist CD24, CTLA4, TNF R, ENPP1, SIRP-α, and urinstatin.

[0252] In certain embodiments, the therapeutic polypeptide is a GIP, GLP-1, and glucagon receptor It contains an agonist in one or more parts of the body. GIP is a peptide consisting of 42 amino acids. It is a thiocyanide that stimulates insulin secretion from pancreatic beta cells in the presence of glucose. It is an incretin that plays a physiological role in glucose homeostasis. GLP-1 It is a peptide consisting of 36 amino acids that stimulates glucose-dependent insulin secretion. It is an incretin that has been shown to prevent hyperglycemia in diabetic patients. GLP-1 The main biologically active fragment is a 30-amino acid C-terminal amidated peptide (GLP). -1 7-36 It is produced as a peptide consisting of 29 amino acids. It binds to and activates glucagon receptors on liver cells, triggering a process called glycogenolysis. By releasing glucose stored in the form of glycogen into the liver through the sac, It helps maintain blood glucose levels.

[0253] In certain embodiments, the exemplary compounds of this disclosure are GIP receptors and GLP-1 receptors. This includes a dual agonist in some other embodiments of the present disclosure. The compound contains a triple agonist for GIP, GLP-1, and the glucagon receptor.

[0254] This opening has activity in one or more of the following: GIP, GLP-1, and / or glucagon receptor. The compounds shown are associated with T2DM, obesity, dyslipidemia, and cardiovascular disease (C). VD), major adverse cardiovascular events (MACE), chronic kidney Diseases (chronic kidney disease, CKD), heart failure (HF), hypertension (hypertension, HT), peripheral artery disease (PAD), closure Obstructive sleep apnea (OSA), metabolic syndrome, 2 Type 2 diabetes mellitus (T2DM), blood glucose control, osteoarthritis (OA), chronic lower back pain ( Chronic lower back pain (CLBP), neurodegenerative disorders and / or cognitive impairment, e.g., Al Alzheimer's disease (AD), Parkinson's disease, metabolic steatohepatitis (ME) Tabolic steatohepatitis (MASH), cirrhosis, fatty liver disease (FASH) LD), polycystic ovary syndrome and / or alcohol abuse disorder, NAFLD and NASH, Many conditions and diseases, including but not limited to metabolic syndrome and bone-related disorders. It may have therapeutic value in the disease or disorder. For example, Jall et al. (201 7) Mol.Metab.6:440-446, Carbone et al. (201 6) J. Gastroenterol. Hepatol. 31:23-31, Finan et al.(2016)Trends Mol.Med.22:359-376, C hoi et al. (2017) Potent body weight loss and efficacy in a NASH animal model by a novel long-acting GLP-1 / Glucagon / GIP tr iple-agonist(HM15211),ADA Poster 1139-P, Ding (2008) J. Bone Miner. Res. 23:536-543, Ta i et al. (2018) Brain Res. 1678:64-74, Mulle r et al. (2017)Physiol.Rev.97:721-766, Fin an et al.(2013)Sci.Transl.Med.5:209, Hols Refer to cher (2014) Biochem. Soc. Trans. 42:593-600. I want to be illuminated.

[0255] In certain embodiments, the therapeutic polypeptide is X1X2X3X4TX6TSDX 10 X 11 X 12 X 13 LX 15 X 16 KAX 19 X 20 X 21 FIX 24 X 25 LX 27 X 28 X 29 X 30 X 31 X 32 X 33 X34 X3 5X 36 X 37 X 38 X 39 X 40 X 41 (Sequence ID 1218) X1 is H, NMeY, or Y. X2 is Ac4c, Aib, αMeS, Iva, or D-Ala. X3 is Q, E, or H. X4 is G or D-Ala, X6 is αMeF(2F), F, or αMeF. X 10 is 4-Pal, Y, or V, X 11 is Aib, S, or αMeS, X 12 is I or S, X 13 It is L or αMeL, X 15 It is either D or E, X 16 is K, E, or Orn, X 19 is A or Q, X 20 These are Aib, αMeL, Iva, or αMe4Pal. X 21 is E, Q, D, or Orn, X 24 These are K, Q, E, D-Glu, or D-Gln. X 25 is αMeY, W, or Y, X 27 is I, L, or V, X 28 is K, E, or A, X 29 is Aib, G, or Q, X 30 is G or S, X 31 is P, G, E, or Orn, X 32It either does not exist, or it is S or P. X 32 If it is S or P, then X 33 is S, X 33 If X is S, 34 is G or Aib, X 34 If X is G or Aib, 35 It does not exist, or it is A or Orn. the law of nature, X 35 If X is A or Orn, 36 It does not exist, or it is P. X 36 If P, then X 37 It does not exist, or it is P. X 37 If P, then X 38 It does not exist, or it is P. X 38 If P, then X 39 It does not exist, or it is S, Orn, or G. X 39 If is S, Orn, or G, then X 40 It does not exist, or it is K or G. X 40 If it is K or G, then X 41 It does not exist, or it is S or G. X 32 If X does not exist, 33 ~X 41 It does not exist, X 35 If X does not exist, 36 ~X 41 It does not exist, X 36 If X does not exist, 37 ~X 41 It does not exist, X 37 If X does not exist, 38 ~X 41 It does not exist, X 38 If X does not exist, 39 ~X 41 It does not exist, X 39 If X does not exist, 40 and X 41 It does not exist, X 40 If X does not exist, 41 It does not exist.

[0256] In certain embodiments, the therapeutic polypeptide is sequence number 1 to sequence number 186 or sequence number Contains 90% sequence identity with any of sequence numbers 996 to 1038. Specific implementation In terms of form, therapeutic polypeptides are sequence numbers 1 to 186 or sequence numbers 996 to Contains one of the column numbers 1038.

[0257] In certain embodiments, the therapeutic polypeptide is YAibEGTX6TSDX 10 X 11 X 12 X 13 LX 15 X 16 KAQX 20 X2 1FIX 24 X 25 LX 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 X 40 X 41 (Sequence ID 1219) X6 is αMeF(2F), F, or αMeF. X 10 is 4-Pal, Y, or V, X 11 is Aib or S, X 12 is I or S, X 13It is L or αMeL, X 15 It is either D or E, X 16 is E or Orn, X 20 These are Aib, αMeL, or Iva, X 21 is E, Q, D, or Orn, X 24 These are K, Q, E, or D-Glu, X 25 is αMeY or Y, X 27 is I, L, or V, X 28 is K, E, or A, X 29 is Aib, G, or Q, X 30 is G or S, X 31 is P, G, E, or Orn, X 32 It either does not exist, or it is S or P. X 32 If it is S or P, then X 33 is S, X 33 If X is S, 34 is G or Aib, X 34 If X is G or Aib, 35 It does not exist, or it is A or Orn. X 35 If X is A or Orn, 36 It does not exist, or it is P. X 36 If P, then X 37 It does not exist, or it is P. X 37 If P, then X 38 It does not exist, or it is P. X 38 If P, then X 39 It does not exist, or it is S, Orn, or G. X 39If is S, Orn, or G, then X 40 It does not exist, or it is K or G. X 40 If it is K or G, then X 41 It does not exist, or it is S or G. X 32 If X does not exist, 33 ~X 41 It does not exist, X 35 If X does not exist, 36 ~X 41 It does not exist, X 36 If X does not exist, 37 ~X 41 It does not exist, X 37 If X does not exist, 38 ~X 41 It does not exist, X 38 If X does not exist, 39 ~X 41 It does not exist, X 39 If X does not exist, 40 and X 41 It does not exist, X 40 If X does not exist, 41 It does not exist.

[0258] In some embodiments, the polypeptide of SEQ ID NO: 1219 is used for GIP receptors and GL receptors. It has dual agonist activity at the P-1 receptor. In some embodiments, polypept Chido includes at least one of the following: X 10 Y is X 11 It is Aib ri, X 24 E is X 30 is S. In some embodiments, polypeptide It includes at least two of the following: X 10 Y is X 11 Aib is, X 24 E is X 30is S. In some embodiments, the polypeptide is Includes each of the following: X 10 Y is X 11 Aib and X 24 is E Yes, X 30 S is.

[0259] In some embodiments, the polypeptide further comprises at least one of the following: X6 is αMeF(2F), and X 12 is I and X 13 αMeL and X 15 D is X 16 Orn and X 20 Aib and X 21 E or Q is X 25 αMeY and X 27 is I and X 28 is K or E ri, X 29 is Aib or G, and X 31 is P or G, and X 32 It does not exist. or S, X 40 is K. In some embodiments, the polypeptide is This includes each of the following: X6 is αMeF(2F), and X 12 is I and X 13 teeth , αMeL, X 15 D is X 16 Orn and X 20 In Aib Yes, X 21 is E or Q, and X 25 αMeY and X 27 is I and X 28 is K or E, and X 29 is Aib or G, and X 31 is P or G, X 32It does not exist, or it is S, X 40 is K. In some embodiments, , X 21 E is X 28 E is X 29 G is X 31 P is , X 32 S is.

[0260] In a particular embodiment, the therapeutic polypeptide comprises each of the following: X1 is Y X2 is Aib, X3 is E, X4 is G, and X6 is αMeF( 2F) and X 10 is 4-Pal or Y, and X 11 is Aib or S, and X 12 is I and X 13 αMeL and X 15 D is X 16 is E or Orn is, X1 is Q, and X 20 Aib and X 21 is E, and X2 4 is Q, E, or D-Glu, and X 25 αMeY and X 27 is I , X 28 E is X 29 is Aib or G, and X 30 is G or S, X 31 P is X 32 is G or S, X 32 If X is S, 33 is S ri, X 34 G is X 35 A is X 36 P is X 37 is P ri, X 38 P is X 39 S is X 40 is K, and X41 It does not exist. , X 32 If X does not exist, 33 ~X 41 It does not exist. In certain embodiments, therapeutic The polypeptide contains at least one of the following: X 10 is Y or 4-Pal ri, X 11 is Aib or S, and X 24 is Q, E, or d-Glu, and X 30 is G or S. In certain embodiments, the therapeutic polypeptide comprises each of the following: Hmm. X 10 is Y or 4-Pal, and X 11 Aib and X 24 E is , X 30 S is X 40 It is K. In some embodiments, the above amino acids One or more of the modifications result in a therapeutic polypeptide with improved proteolytic stability. In some embodiments, one or more of the above amino acid modifications are GIP and / Alternatively, it may result in a therapeutic polypeptide with increased activity at the GLP-1 receptor.

[0261] In some embodiments, the therapeutic polypeptide is sequence number 1 to sequence number 92 or sequence number Contains 90% sequence identity with any of sequence numbers 996 to 999. In this embodiment, the therapeutic polypeptide is SEQ ID NOs: 1 to 92 or SEQ ID NOs: 996 Contains one or more conservative amino acid substitutions for any of the sequence numbers 999. In that embodiment, the therapeutic polypeptide is SEQ ID NOs: 1 to 92 or SEQ ID NOs: 99 For any of the sequences 6 through 999, 5 or fewer, 4 or fewer, 3 or fewer, or 2 or fewer. Includes one or fewer conservative amino acid substitutions. In some embodiments, therapeutic polypeptide Tide is one of the following: SEQ ID NOs: 1-92 or SEQ ID NOs: 996-999 This includes. In some embodiments, the therapeutic polypeptide is SEQ ID NOs: 1, 2, 22, 4 Any of 3, 45, 48, 55, 56, 58, 59, 61, 66, 68, and 47 It includes ka.

[0262] In other embodiments, the therapeutic polypeptide is X1X2X3X4TX6TSDX 10 X 11 X 12 X 13 LX 15 X 16 KAQX 20 X 21 FIX 24 X 25 LX 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 X 40 X 41 (Sequence ID 1220) X1 is H, NMeY, or Y. X2 is Ac4c, Aib, αMeS, Iva, or D-Ala. X3 is Q, E, or H. X4 is G or D-Ala, X6 is αMeF(2F), F, or αMeF. X 10 is 4-Pal, Y, or V, X 11 is Aib, S, or αMeS, X 12 is I or S, X 13 It is L or αMeL, X 15 It is either D or E, X 16 is E or Orn, X 20 These are Aib, αMeL, Iva, or αMe4Pal. X 21 is E, Q, D, or Orn, X 24 These are K, Q, E, D-Glu, or D-Gln. X 25 is αMeY or Y, X 27 is I, L, or V, X 28 is K, E, or A, X 29 is Aib, G, or Q, X 30 is G or S, X 31 is P, G, E, or Orn, X 32 It either does not exist, or it is S or P. X 32 If it is S or P, then X 33 is S, X 33 If X is S, 34 is G or Aib, X 34 If X is G or Aib, 35 It does not exist, or it is A or Orn. the law of nature, X 35 If X is A or Orn, 36 It does not exist, or it is P. X 36 If P, then X 37 It does not exist, or it is P. X 37 If P, then X 38 It does not exist, or it is P. X 38 If P, then X39 It does not exist, or it is S, Orn, or G. X 39 If is S, Orn, or G, then X 40 It does not exist, or it is K or G. X 40 If it is K or G, then X 41 It does not exist, or it is S or G. X 32 If X does not exist, 33 ~X 41 It does not exist, X 35 If X does not exist, 36 ~X 41 It does not exist, X 36 If X does not exist, 37 ~X 41 It does not exist, X 37 If X does not exist, 38 ~X 41 It does not exist, X 38 If X does not exist, 39 ~X 41 It does not exist, X 39 If X does not exist, 40 and X 41 It does not exist, X 40 If X does not exist, 41 It does not exist.

[0263] In certain embodiments, the therapeutic polypeptide of SEQ ID NO: 1220 is GIP, GLP-1 and has triagonist activity at the glucagon receptor. In some embodiments, Therapeutic polypeptides are X 10 Y is X 24 E is X 30 S is X4 The condition includes at least one of which is 0 = K. In some embodiments, therapeutic poly Peptides are X 10 Y is X 24 E is X30 S is X 40 It is K This includes each of the things mentioned above.

[0264] In some embodiments, the therapeutic polypeptide comprises at least one of the following: X1 is H or Y, X2 is Ac4c or Aib, X3 is Q, X 4 is G, X6 is αMeF(2F), and X 11 It is αMeS, and X 12 is I and X 13 αMeL and X 15 D is X 16 It is Orn ri, X 20 It is αMe4Pal, and X 21 is E or Orn, and X 25 , αM eY and X 27 is I and X 28 E is X 29 is Aib or G , X 31 P is X 32 S is X 33 S is X 34 G is , X 35 A is X 36 P is X 37 P is X 38 P is , X 39 is S. In some embodiments, the therapeutic polypeptide is one of the following This includes X1 being H or Y, X2 being Ac4c or Aib, and X3 being Q. Yes, X4 is G, X6 is αMeF(2F), X 11 It is αMeS. , X 12 is I and X 13 αMeL and X 15 D is X 16 is, O rn and X 20 It is αMe4Pal, and X 21 is E or Orn, and X 25 αMeY and X 27 is I and X 28 E is X 29 is Aib or G and X 31 P is X 32 S is X 33 S is X 34 teeth, G and X 35 A is X 36 P is X 37 P is X 38 teeth, P is X 39 is S. In some embodiments, X1 is Y and X2 is Aib is Q, X3 is Q, X 21 E is X 29 It is G.

[0265] In some embodiments, the therapeutic polypeptide comprises each of the following: X1 is H Or Y, X2 is Aib, X3 is Q, X4 is G, X6 is αMeF(2F), X 10 is Y or 4-Pal, and X 11 is S or αMe S and X 12 is I and X 13 αMeL and X 15 D is X2 0 is Aib or αMe4Pal, and X 21 is E or Orn, and X 24 is, E or d-Glu, X 25 αMeY and X 27 is L or I, and X 28 E is X 29 ~X 39is GSPSSGAPPPS, and X 40 is K In some embodiments, the therapeutic polypeptide comprises at least one of the following: . X 10 is Y or 4-Pal, and X 24 is E or d-Glu, and X 30 teeth, S. In some embodiments, the therapeutic polypeptide comprises each of the following: X 10 is Y or 4-Pal, and X 24 is E or d-Glu, and X 30 is S Yes, X 40 It is K, and it is conjugated to the linker.

[0266] In some embodiments, one or more of the above amino acid modifications improve protein degradation stability. This yields improved therapeutic polypeptides. In some embodiments, the above amino acid modification One or more of the ornaments are therapeutic polypeptides with increased activity at GIP and / or GLP-1 receptors. It brings about chid.

[0267] In some embodiments, the therapeutic polypeptide is SEQ ID NOs: 93-186 or SEQ ID NOs: Includes 90% sequence identity with any of 1000-1038. Several embodiments So, therapeutic polypeptides are sequence numbers 93-186 or 1000-1038. This includes one or more conservative amino acid substitutions for any of the following embodiments. In some embodiments, The therapeutic polypeptide is one of the following: SEQ ID NOs: 93-186 or 1000-1038 For any of the following, 5 or fewer, 4 or fewer, 3 or fewer, 2 or fewer, or 1 or fewer conservative A Includes mino acid substitution. In some embodiments, the therapeutic polypeptide is SEQ ID NOs: 93-1 Includes either 86 or one of sequence numbers 1000-1038. In some embodiments, Therapeutic polypeptides include SEQ ID NOs: 97, 113, 128, 130, 144, 171, 1 Includes one of the following 83.

[0268] The structural features of the compounds described in specific embodiments herein relate to the activity at each receptor. To obtain the desired effect, each of the odors of GIP, GLP-1, and glucagon receptors is treated. It has appropriate activity (i.e., triple agonist activity), but activity at the other two receptors In a situation where it is administered in a dose that overwhelms or is sufficient to produce activity in all three receptors In any one receptor, either to cause undesirable side effects. The goal is to limit its activity to a moderate level.

[0269] The polypeptides described herein to GIP, GLP-1, and glucagon receptors The affinity of the cydide is determined using techniques known in the art to measure receptor binding levels. It can be measured and is generally expressed as an inhibition constant (Ki) value. The activity of the polypeptides described in this specification is also known in the art, for example, It can be measured using techniques including the in vitro activity assays described below, and generally, effective concentration effective concentration 50, EC 50 This is expressed as a value, which is the dose-response curve. This is the concentration of the compound that causes the simulation to reach a maximum of half the amount shown by the line.

[0270] In some embodiments, the polypeptides described herein are GLP-1 receptors. It is a partial agonist in the body, which is the HEK293 cell GL described herein. If demonstrated by the P-1 receptor internal distribution assay, natural GLP-1 7~36 (array It shows agonism of 80% or less compared to (number 1221). In other embodiments, the present The polypeptides described in the document are complete agonists of the GLP-1 receptor, and This was determined by the HEK293 cell GLP-1 receptor internal delivery assay described herein. If proven, natural GLP-1 7~36 Compared to (Sequence ID 1221), it is more than 80% It exhibits agonism. In some embodiments, the polypeptides described herein are Natural glucagon (SEQ ID NO: 1222), GIP (SEQ ID NO: 1223), and GLP-17 -36 Compared to (SEQ ID NO: 1221), glucagon, GIP and GLP-1 receptor vesicles It has a greater effect in one or more of the following ways.

[0271] In addition to the sequences described herein, one or more polypeptides described herein The polypeptide may include the conservative amino acid substitutions of GIP, GLP-1, and glucans. Provided that it remains possible to bind to and activate GON receptors. .

[0272] Due to the structural features of specific embodiments of the compounds described herein, the compounds are neutral p Improved solubility of analogs in aqueous solutions near H, improved chemical and physical formulation stability, immersion Improved peptide membrane permeability in the presence of permeability enhancers, expanded pharmacokinetic profile, and injection This relates to the potential for development as a therapeutic treatment, including minimizing the possibility of site reactions or immunogenicity. It will also possess many other beneficial attributes.

[0273] The beneficial combinations of features of the exemplary analogues described herein are not applicable to any single individual. Rather than being the result of a single modification, it is a novel combination of structural features described herein. Please note that this is achieved through [method / method].

[0274] In some embodiments, the polypeptides described herein are amidated In some embodiments, the polypeptides described herein have a C-terminal group It has a modification, and the modification is either NH2 or absent. In some embodiments, Polypeptides described herein have an OH group at the C-terminus. Exemplary therapeutic polypeptides A non-restrictive list of petit des is provided in Table 1 below.

[0275] [Table 11-1]

[0276] [Table 11-2]

[0277] [Table 11-3]

[0278] [Table 11-4]

[0279] [Table 11-5]

[0280] [Table 11-6]

[0281] [Table 11-7]

[0282] [Table 11-8]

[0283] [Table 11-9]

[0284] Acylation of therapeutic polypeptides As previously mentioned, in some embodiments, certain polypeptides described herein The 'd' is a natural or non-natural amino acid that has a functional group that can be used for conjugation, for example. For example, albumin such as fatty acid portions that are directly bound or conjugated by linkers. It includes the bonding portion. Such conjugations are sometimes called acylation. Depending on the combination, amino acids that have functional groups usable for conjugation are K, C, and E. , and D. A specific example is having a functional group that can be used for conjugation. The amino acid is K, and the conjugation is directed to the epsilon-amino group of the K side chain. That is the case.

[0285] In some embodiments, the polypeptides described herein are functionalized with amino acids. C is chemically conjugated to the base either via a direct bond or via a linker. 16 ~C2 2. Fatty acids are used. The length and composition of fatty acids are used in relation to the half-life of polypeptides and animal models. This affects their efficacy in the body, as well as their solubility and stability. 16 ~C 22 Conjugation to a saturated mono- or di-fatty acid results in a polypeptide that exhibits a desirable half-life, desirable in vivo efficacy in animal models and desirable solubility and stability characteristics in vivo.

[0286] Examples of saturated C 16 ~C 22 fatty acids for use herein include, but are not limited to, palmitic acid (hexa decanoic acid) (C 16 monoacid), hexadecanedioic acid (C 16 diacid), margaric acid (hepta decanoic acid) (C 17 monoacid), heptadecanedioic acid (C 17 diacid), stearic acid (C 18 monoacid), octadecanedioic acid (C 18 diacid), nonadecylic acid (nonadecanoic acid) (C 19 monoacid ), nonadecanedioic acid (C 19 diacid), arachidic acid (eicosanoic acid) (C 20 monoacid), eic osanedioic acid (C 20 diacid), heneicosylic acid (heneicosanoic acid) (C 21 monoacid), heneicosanedioic acid (C 21 diacid), behenic acid (docosanoic acid) (C 22 monoacid), docosan edioic acid (C 22 diacid), and include, but are not limited to, branched and substituted derivatives thereof .

[0287] In some cases, the C 16 ~C 22 fatty acid is a saturated C 18 monoacid, a saturated C 18 diacid, a saturated C 19 monoacid, a saturated C 19 diacid, a saturated C 20 monoacid, a saturated C 20 diacid, and branched and may be a substituted derivative. In a more specific example, C 16 ~C 22 fatty acid is octadecaned ioic acid (C 18 diacid) or eicosanedioic acid (C 20 diacid).

[0288] In some embodiments, the lysine at position 17 is conjugated to a fatty acid. In some embodiments, the fatty acid is C 16 ~C 22 fatty acid.

[0289] In some embodiments, the therapeutic polypeptide is conjugated to the fatty acid via a linker . In some embodiments, the linker comprises 1 to 5 amino acids. In an exam ple where the linker comprises at least one amino acid, the amino acid may be 1 to 5 Glu or γGlu amino acid residues. In some examples, the linker can comprise 1, 2 or 3 or 4 or 5 Glu or γGlu amino acid residues, and the Glu or γGl u amino acid residues include the D form thereof. For example, the linker comprises any one of 1, 2, 3 or 4 γGlu amino acid residues. Alternatively, the linker may comprise 1 ~5 (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl) ("AEEA ") and / or 1 to 5 amino acid residues used in combination with 1 to 5 εK moieties (e.g., Glu or γGlu amino acids, etc.). Specifically, the link er may be a combination of 1 to 5 Glu or γGlu amino acids and 1 to 5 (2-[2-(2-amin oethoxy)-ethoxy]-acetyl) moieties, or a combination of 1 to 5 Glu or γGlu amino acids and 1 to 5 εK moieties. In some examples, The linker consists of one, two, or three γGlu amino acids and one or two (2-[2- It is a combination of (2-amino-ethoxy)-ethoxy]-acetyl) or the εK moiety. obtain.

[0290] For example, in some embodiments, the polypeptides described herein are as follows: formula: (γGlu) a -(2-[2-(2-amino-ethoxy)-ethoxy]-acetyl) b -(γGlu) c -CO-(CH2) p Linkers and fatty acid formation having the -CO2H structure The formula has minutes, where a is 0, 1, or 2, b is 0, 1, or 2, and c is 0 The integer is 1, 2, or 3, and p is an integer between 14 and 20.

[0291] In some preferred embodiments, a is 0 or 1, and b is 0, 1, or 2 The equation is such that c is 1, 2, or 3, and p is an integer between 14 and 20.

[0292] In some embodiments, a is 0, b is 1, and c is 1 or 2. Therefore, p is either 16 or 18.

[0293] For example, in some embodiments, a is 0, b is 1, and c is 1. Yes, p is 16, and its structure is shown below.

[0294] [ka]

[0295] For example, in some embodiments, a is 0, b is 1, and c is 1. Yes, p is 18, and its structure is shown below.

[0296] [ka]

[0297] In some embodiments, a is 0, b is 1, c is 2, and p It is 16, and its structure is shown below.

[0298] [ka]

[0299] In some embodiments, a is 0, b is 1, c is 2, and p It is 18, and its structure is shown below.

[0300] [ka]

[0301] In some embodiments, a is 0, b is 2, c is 1, and p It is 16 or 18.

[0302] For example, in some embodiments, a is 0, b is 2, and c is 1. Yes, p is 16, and its structure is shown below.

[0303] [ka]

[0304] In some embodiments, a is 0, b is 2, c is 1, and p It is 18, and its structure is shown below.

[0305] [ka]

[0306] In some embodiments, a is 0, b is 0, c is 2, and p It is 16 or 18.

[0307] For example, in some embodiments, a is 0, b is 0, and c is 2. Yes, p is 16, and its structure is shown below.

[0308] [ka]

[0309] In some embodiments, a is 0, b is 0, c is 2, and p It is 18, and its structure is shown below.

[0310] [ka]

[0311] In some embodiments, a is 0, b is 0, c is 3, and p It is 16 or 18.

[0312] For example, in some embodiments, a is 0, b is 0, and c is 3. Yes, p is 16, and its structure is shown below.

[0313] [ka]

[0314] In some embodiments, a is 0, b is 0, c is 3, and p It is 18, and its structure is shown below.

[0315] [ka]

[0316] In some embodiments, a is 1, b is 1, c is 1, and p It is 16 or 18.

[0317] For example, in some embodiments, a is 1, b is 1, and c is 1. Yes, p is 16, and its structure is shown below.

[0318] [ka]

[0319] For example, in some embodiments, a is 1, b is 1, and c is 1. Yes, p is 18, and its structure is shown below.

[0320] [ka]

[0321] In some embodiments, the polypeptide described herein is of the following formula: (γGlu) d -(εK) e -(γGlu) f -CO-(CH2) q -CO2H structure It has a linker and a fatty acid component, where d is 0, 1 or 2, and e is 0, 1 , or 2, f is 0, 1, 2 or 3, and q is an integer between 14 and 20.

[0322] For example, in one embodiment, d is 0, e is 2, f is 1, and q is an integer between 14 and 20. In some embodiments, d is 0 and e is 2 Therefore, f is 1, and q is 16 or 18.

[0323] For example, in some embodiments, d is 0, e is 2, and f is 1. Yes, q is 16, and its structure is shown below.

[0324] [ka]

[0325] For example, in some embodiments, d is 0, e is 2, and f is 1. Yes, q is 18, and its structure is shown below.

[0326] [ka]

[0327] A non-exclusive list of exemplary acylation therapeutic polypeptides is provided in Table 2 below.

[0328] [Table 12-1]

[0329] [Table 12-2]

[0330] [Table 12-3]

[0331] [Table 12-4]

[0332] Table 12-5

[0333] Table 12-6

[0334] Table 12-7

[0335] Table 12-8

[0336] Table 12-9

[0337] Table 12-10

[0338] Table 12-11

[0339] Table 12-12

[0340] Table 12-13

[0341] Table 12-14

[0342] [Table 12-15]

[0343] [Table 12-16]

[0344] [Table 12-17]

[0345] [Table 12-18]

[0346] [Table 12-19]

[0347] [Table 12-20]

[0348] [Table 12-21]

[0349] Other definitions and abbreviations The amino acid sequences of therapeutic polypeptides described herein are typically standard one-sentence sequences. Naturally occurring mycelium, as shown herein using the letter code (e.g., L = leucine). α-methyl substituted residues of natural amino acids (e.g., α-methyl-leucine (αM) (eL), and certain other non-natural amino acids such as alpha-aminoisobutyric acid (Aib) are incorporated. The structures of these amino acids are shown below.

[0350] [ka]

[0351] [ka]

[0352] As used herein, "Orn" means L-ornithine. When used, "4-Pal" or "4Pal" refers to 3-(4-pyridyl)-L-alani This refers to (S)-2-amino-3-(pyridine-4-yl)propanoic acid. When used in writing, "3-Pal" or "3Pal" means 3-(3-pyridyl)-L- This refers to alanine or (S)-2-amino-3-(pyridine-3-yl)propanoic acid. As used herein, "αMe-4-Pal" or "αMe4Pal" means alpha This refers to α-methyl-3-(4-pyridyl)-L-alanine. In this specification, "αMeY" stands for alpha-methyl-L-tyrosine. In this context, "αMeL" means alpha-methyl-leucine. In this specification, "Ac3c" means 1-aminocyclopropanecarboxylic acid. When used, "Ac4c" means 1-aminocyclobutane-1-carboxylic acid. As used herein, "D-Ala" and "a" refer to D-alanine, respectively. Tastes. As used herein, "D-Glu" and "e" respectively refer to D-glutamic acid. It means amino acid. As used herein, "Aib" means 2-aminoisobutyric acid. Tastes good. As used herein, "NMeY" means N-methyl-tyrosine. As used herein, "Dap" refers to (S)-2,3-diaminopropanoic acid. Tastes good. As used herein, "Dab" refers to (S)-2,4-diaminobutanoic acid. This means. As used herein, "Hyp" means hydroxy-L-proline. To taste. Where used herein, "K(Ac)" means N 6 -acetyl-L-lysine Meaning. As used herein, "γGlu" means gamma-L-glutamic acid. In this specification, "Aad" refers to (S)-2-aminohexanediic acid. Tastes. As used herein, "F(4CN)" refers to 4-cyano-L-phenyl This refers to ranine or (S)-2-amino-3-(4-cyanophenyl)propanoic acid. When used in a specification, "F(4NO2)" means 4-nitro-L-phenylalanine or (S)-2-amino-3-(4-nitrophenyl)propanoic acid. In this specification Where used, "αMeS" means alpha-methyl-L-serine. When used in this context, "αMeF" means alpha-methyl-L-phenylalanine. As used herein, "αMeF(2F)" means alpha-methyl-2-full. Oro-L-phenylalanine or (S)-2-amino-3-(2-fluorophenyl)- This refers to 2-methylpropanoic acid. When used herein, "L-Iva" and "I "va" means L-isovalin. When used herein, "D-Gln" and Each "q" represents D-glutamine.

[0353] The specific abbreviations used herein are defined as follows: "AcOH" stands for acetic acid. "ACN" refers to acetonitrile, and "BEA" refers to 4-(bis(2-((( (9H-Fluoren-9-yl)methoxy)carbonyl)amino)ethyl)amino)-4 - refers to oxobutanoic acid, and "Boc" refers to tert-butoxycarbonyl, and "t- "Bu" refers to tert-butyl, and "DABA" refers to 3,5-diaminobenzoic acid. "DCM" refers to dichloromethane, and "DMAP" refers to 4-dimethylaminopyridine. "DIC" refers to diisopropylcarbodiimide, and "Et2O" refers to diethyl "DIEA" refers to diisopropylethylamine, and "DMF" refers to diisopropyl ether. , refers to dimethylformamide, and "DMAP" refers to 4-dimethylaminopyridine, "ivDde" is 1-(4,4-dimethyl-2,6-dioxocyclohex-1-ylide It refers to (n)-3-methylbutyl, "EtOH" refers to ethanol, and "Â" is "Ethyl acetate" refers to ethyl acetate, "eq" refers to equivalent weight, and "Fmoc" refers to fluorenylmethyl ethyl acetate. "Xycarbonyl" refers to the xycarbonyl molecule, while "FPLC" refers to high-performance protein liquid chromatography. "HFIP" refers to hexafluoroisopropanol, and "IPA" refers to isopropyl It refers to an alcohol, and "MalDab" is (S)-2-((tert-butoxycarb (Nyl)amino)-4-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-i (S)-3-((tert-butoxycarb (Nyl)amino)-2-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-i (L) refers to propanoic acid, and β-MalDap is (S)-2-((tert-butoxycal Bonyl)amino)-3-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1- yl)propanoic acid, "MeOH" refers to methanol, "min" refers to minute(s) / minute(s), "MTT" refers to methylthiazolium tetrazolium, "M tt" refers to 4-methyltrityl, "Oxyma" refers to ethyl cyanohydroxyiminoacetate yl, "PEG" refers to polyethylene glycol, "PyBOP" refers to benzo triazol-1-yl-oxy-tris-pyrrolidino-phosphonium hexafluoro phosphate, "RT (room temperature)" refers to room temperature, "t-Bu" refers to tert-butyl, "SPPS (solid-phase peptide synthesis)" refers to solid-phase peptide synthesis, "soln" refers to solution, "TFA" refers to trifluoroacetic acid, "TIPS" refers to triisopropylsilane, "TCEP" refers to tris(2-carboxy ethyl)phosphine hydrochloride, and "THF" refers to tetrahydrofuran.

[0354] As used herein, "BEA-MalDap2" refers to the structure below.

[0355]

Chemical Formula

[0356] As used herein, "dCAP" refers to the structure below.

[0357]

Chemical Formula

[0358] As used herein, "BEA-MSTP2" refers to the structure below.

[0359] [ka]

[0360] As used herein, "BEA-acetyl-2" refers to the following structure:

[0361] [ka]

[0362] As used herein, "BEA-OD2" refers to the following structure:

[0363] [ka]

[0364] As used herein, "BEA-β-MalDap2" refers to the following structure:

[0365] [ka]

[0366] As used herein, "about" means, for example, the stated concentration, length, molecular weight, p Within a statistically significant range of one or more values ​​such as H, sequence identity, time frame, temperature, or volume. This means that such a value or range is typically within 20% of a given value or range, or more accurately, within 20% of the given value or range. In terms of type, it can be within 10%, and more typically within 5%. The permissible variations included depend on the specific system in the study and are easily understood by those skilled in the art. It is possible.

[0367] When used herein, one of GIP, GLP-1, or glucagon receptor In reference to the above, terms such as "activate," "to activate," and "being activated" refer to the technology in question. Measurements were taken using assays known in the field, for example, the in vitro assays described below. In this case, the ability of a compound to bind to a receptor and induce a response at the receptor, for example, this specification This refers to the capabilities of the polypeptide as described in the book.

[0368] As used herein, "amino having functional groups available for conjugation" "Acid" has a functional group that can be directly attached to a fatty acid or conjugated by a linker, for example. This refers to any natural (encoded) or non-natural (non-encoded) amino acid. Examples of functional groups include alkynyl, alkenyl, amino, azide, bromo, and carboxyl. Examples include, but are not limited to, chloro, iodine, and thiol groups. Examples of natural amino acids containing functional groups include K (amino), C (thiol), and E (carbococcus). Examples include sil (sil) and D (carboxyl).

[0369] As used herein, "conservative amino acid substitution" refers to a property (e.g., charge, side chain) The properties (such as hydrophobicity / hydrophilicity, skeletal structure, and rigidity) are similar, and the resulting substitutions are similar. Amino acids that have minimal impact on the biological activity of peptides or polypeptides This refers to acid substitution. Conservative substitutions of functionally similar amino acids are well known in the relevant field. Therefore, it is not necessary to explain it comprehensively in this specification.

[0370] When used herein, "C 16 ~C 22 "Fatty acids" are composed of 16 to 22 carbon atoms. This means a carboxylic acid containing. C is suitable for use in this specification. 16 ~C 22 Fatty acids are saturated It may be a monoacid or a saturated diacid. As used herein, “saturated” means that the fatty acid is carbon - This means that the carbon double or triple bond is not present. Unless otherwise specified, this specification When referred to as such, fatty acids may also contain other acidic groups such as phosphonic acids or sulfonic acids.

[0371] As used herein, “effective amount” means a single or multiple doses to an individual requiring it. This specification provides the desired effect to such individuals under diagnosis or treatment after administration of a single dose. The amount, concentration, or one or more polypeptides or pharmaceutically acceptable salts thereof described in the book. This refers to the dosage. The effective dose is the result obtained by using known techniques and under similar circumstances. The effective amount for an individual can be easily determined by observing the results. When determining the species of mammal; its size, age, and overall health; and the specific disease involved. Affected or disabled; degree or involvement or severity of the disease or disability; individual patient response; specific treatment administered Specific polypeptide; mode of administration; bioavailability characteristics of the administered preparation; selected dose regimen This includes, but is not limited to, several, menstruation; concomitant use of medications; and other related circumstances. Those factors will be taken into consideration.

[0372] As used herein, “extended duration of action” refers to the binding parent of the polypeptide. Its compatibility and activity persist for a longer period than natural human GIP, GLP-1, and glucagon. At least once a day, three times a week, twice a week, once a week, every other week (every two weeks), or once a month (low frequency) This means it can be administered at a dose. The polypeptide's time-acting profile is These can be measured using known pharmacokinetic testing methods, such as those used in the following examples.

[0373] As used herein, “polypeptide” or “peptide” refers to a combination of elements in a specific sequence. This refers to a polymer composed of multiple bonded amino acid residues. These amino acids are They are linked via an amide (peptide) bond, which means that the carboxyl group of one amino acid is connected to the amide (peptide) bond. It is formed when it reacts with the amino group of another amino acid and releases a water molecule. This term is derived from the word "heaven." Polymers containing naturally occurring amino acids, and polymers containing one or more amino acids not found in nature. It is applied to polymers.

[0374] As used herein, “the individual requiring it” means, for example, those listed herein. Humans who have a medical condition, disease, disorder, or symptom that requires treatment or therapy, including those who are affected by This refers to mammals such as [specific mammals].

[0375] As used herein, this includes "to treat," "treating," and "for the purpose of treating." This involves suppressing, slowing, stopping, or reversing the progression or severity of existing conditions, diseases, disabilities, or symptoms. It means.

[0376] As used herein, "double agonist activity" with respect to a compound means GIP and This refers to compounds that have activity in each of the GLP-1 receptors, and in particular, compounds related to excessive activity. To provide the benefits of receptor agonism while avoiding the associated undesirable side effects. Therefore, it refers to polypeptides that have sufficient activity at each receptor. Furthermore, a double agonist Polypeptides possessing activating properties were extended in the GIP and GLP-1 receptors, respectively. It has a duration of action, and is administered at a low frequency of once a week, twice a month, once a month, or once a quarter. To make it easier to administer medication.

[0377] As used herein, "triple agonist activity" with respect to a compound means GIP, This refers to compounds that have activity in both GLP-1 and the glucagon receptor, and in particular, While avoiding undesirable side effects associated with excessive activity, the benefits of receptor agonism can be leveraged. This refers to polypeptides that have sufficient activity at each receptor to provide a point. Polypeptides having triple agonist activity are GIP, GLP-1, and glucagon receptors. It has an extended duration of action in terms of its dosage, whether once a week, twice a month, once a month, or four times a week. This makes it advantageous to administer medication at a low frequency, such as once every six months.

[0378] As used herein, the term “sequence identity” refers to the degree of similarity between two sequences. This indicates the degree. The degree of sequence identity between two polypeptides is expressed as a percentage, and is as follows: It can be calculated as follows: %array identity=100% * (Number of identical amino acids) / (Shortest) Length of the common array).

[0379] In a particular embodiment of any polypeptide among the formulas described herein, The polypeptide is derived from any one of the polypeptides described herein using an isotope. It is a body or a pharmaceutically acceptable salt thereof. Various technologies recognized in the art It is understood that isotopic derivatives can be prepared using any of these. Isotope derivatives are generally obtained by substituting an isotope-labeled reagent with a non-isotope-labeled reagent. They can be prepared by carrying out the procedures disclosed in the examples described herein. Any polypeptide or pharmaceutically acceptable salt thereof of any of the formulas described herein. In this embodiment, the polypeptide is one of the polypeptides described herein. It is one of the deuterated derivatives.

[0380] In the polypeptide of the present invention, any atom not specifically designated as a particular isotope This is intended to represent any stable isotope of that atom. Unless otherwise specified, And when an atom is specifically designated as "H" or "hydrogen", that atom is in its natural state It is understood that the atom contains hydrogen in its current isotopic composition. Also, unless otherwise specified, the atom is When specifically designated as "D" or "deuterium," that atom is 0.015% deuterium. It is understood that it contains deuterium at a substantially higher abundance than its natural abundance.

[0381] The polypeptides described herein react with some inorganic acids and organic acids / bases. , which can form pharmaceutically acceptable acid / base addition salts. pharmaceutically acceptable salts and they The general techniques for preparation are well known in the art (e.g., Stahl et al.) al., “Handbook of Pharmaceutical Salts:P roperties, Selection and Use,”2 nd Revised See Edition (Wiley-VCH, 2011). For use in this specification Pharmaceutically acceptable salts include sodium salt, potassium salt, trifluoroacetate salt, and salt Examples include salts and / or acetates. Therefore, in some embodiments, polypept pharmaceutically acceptable salt forms of tide are provided herein. In some embodiments, the drug Scientifically acceptable forms are selected from sodium salts or potassium salts. Several implementations In terms of form, pharmaceutically acceptable forms are selected from the group consisting of sodium salts and potassium salts. In some embodiments, the pharmaceutically acceptable salt is the sodium salt.

[0382] In another embodiment, the polypeptide described herein or a pharmaceutically acceptable salt thereof Pharmaceutical compositions comprising a pharmaceutically acceptable carrier, diluent, or excipient are provided herein. Several pharmaceutical compositions and techniques for preparing them are available in the field. It is knowledge. For example, Remington: The Science and Practice Ice of Pharmacy (Troy, Ed., 21 st Edition, Li See ppincott, Williams & Wilkins, 2006. .

[0383] In some embodiments, the pharmaceutical composition is delivered via parenteral routes (e.g., subcutaneous, intravenous, abdominal). It is suitable for administration via intracavitary, intramuscular, or transdermal means. In some embodiments, the pharmaceutical The composition is suitable for oral administration (e.g., tablets, capsules). In some embodiments... The pharmaceutical composition is administered parenterally. In some embodiments, the pharmaceutical composition is It is administered orally.

[0384] This disclosure also includes the combination of polypeptides described herein or pharmaceutically acceptable salts thereof. This specification provides and includes novel intermediates and methods for forming. The polypeptides can be prepared by various techniques known in the art. For example, methods using chemical synthesis are illustrated in the following examples, or biological expression It is to be used. The specific synthesis steps for each of the routes described herein are described herein. The listed polypeptides can be combined with different methods for preparation. Reagents and The starting materials are readily available to those skilled in the art.

[0385] For chemical synthesis, standard manual or automated solid-phase synthesis procedures can be used. For example, an automated peptide synthesizer is, for example, CEM (Charlotte, North Carolina), CSBio (Menlo Park, California), and and Gyros Protein Technologies Inc. (Tucson, It is commercially available from AZ. Reagents for solid-phase synthesis are readily available from commercial suppliers. This involves blocking interfering groups, protecting amino acids during reactions, coupling, deprotection, and unreacted reactions. For amino acid capping, use a solid-phase synthesizer according to the manufacturer's instructions. It is possible.

[0386] The compounds disclosed herein, unless otherwise specified, include all possible stereoisomers and geometric compounds. This includes isomers (e.g., cis / trans) and structural isomers. Such isomers are molecular These may be present due to chiral centers, double bonds, or other stereoelements within the structure. Unless otherwise specified. To the extent that, the nomenclature used and the structure represented are racemic mixtures, individual enantiomers, This includes diastereomers, positional isomers, and regioisomers, as well as mixtures thereof. This is intended. Regardless of their isomer composition, the present invention relates to salts, solvates, and polymorphs. This includes any pharmaceutically acceptable form of these compounds.

[0387] Regarding biological expression, standard recombinant techniques are used to express all or part of the polypeptide. A polynucleotide having a nucleic acid sequence encoding the amino acid sequence is constructed, and this polynucleotide The rheotide is incorporated into a recombinant expression vector, and the vector is expressed in host cells, such as bacteria and yeast. , and can be introduced into mammalian cells to produce polypeptides. For example, Gre en & Sambrook, “Molecular Cloning: A Labor "Cold Spring Harbor Laboratories Manual" See Ory Press, 4th ed., 2012. Polypeptides are mammal Mammal cells, e.g., CHO, NSO, 20 HEK293, BHK, or COS cells In bacterial cells, for example, Escherichia coli (E. coli), Bacillus subtilis (Bacillus subtilis) us subtilis), or Pseudomonas fluorescens ) in; in insect cells; or in fungal or yeast cells (these are, It can be easily produced (cultivated using techniques known in the relevant art). Vectors containing a renucleotide sequence are processed in different, well-known ways depending on the type of cell host. Therefore, it can be transferred into host cells. Various methods of protein purification can be used. Such methods are known in the relevant technical field.

[0388] The compounds described herein are intended to treat various pathological conditions, disorders, diseases, or symptoms. It can be used. Specifically, a method for treating obesity in individuals is provided, and so Such a method involves administering an effective amount of the following substance to an individual requiring such treatment: The process includes at least the step of administering jugate or a pharmaceutically acceptable salt thereof.

[0389] In addition, methods for chronic weight management in individuals are provided, and such methods are For individuals requiring such treatment, an effective amount of the conjugate described herein, This includes at least the step of administering the pharmaceutically acceptable salt.

[0390] In addition, methods for treating type 2 diabetes (T2DM) in individuals are provided, Methods such as those described herein can be used to deliver an effective amount of the treatment to an individual requiring such treatment. The process includes at least the step of administering a conjugate or a pharmaceutically acceptable salt thereof.

[0391] In addition, non-alcoholic fatty liver disease in individuals Methods are provided to treat such cases (NAFLD), and such methods are such treatments To an individual in need of treatment, an effective amount of the conjugate described herein, or its pharmaceutical The process includes at least the step of administering a moderately acceptable salt.

[0392] Additionally, non-alcoholic steatohepatitis in individuals Methods are provided to treat NASH, and such methods do not require such treatment. To the target individual, an effective amount of the conjugate described herein, or its pharmaceutically acceptable... The process includes at least the step of administering the salt to be used.

[0393] In addition, methods for treating dyslipidemia in individuals are provided, and such methods This involves providing an effective amount of the polypeptide described herein to an individual requiring such treatment. Or, the process may include at least the step of administering a pharmaceutically acceptable salt thereof.

[0394] Additionally, methods for treating metabolic syndrome in individuals are provided. Such methods involve administering an effective amount of the method described herein to an individual requiring such treatment. The process includes at least the step of administering a conjugate or a pharmaceutically acceptable salt thereof.

[0395] In addition, methods for treating osteoarthritis (OA) in individuals are provided, Such methods involve administering an effective amount of the C to an individual requiring such treatment, as described herein. The process includes at least the step of administering a jugate or a pharmaceutically acceptable salt thereof.

[0396] Additionally, methods for treating obesity-related sleep apnea (OSA) in individuals are provided. And such a method is to administer an effective amount, as described herein, to an individual requiring such treatment. The process includes at least the step of administering the listed conjugate or a pharmaceutically acceptable salt thereof. nothing.

[0397] Additionally, methods for treating polycystic ovary syndrome (PCOS) in individuals are provided. And such a method is to administer an effective amount, as described herein, to an individual requiring such treatment. The process includes at least the step of administering the listed conjugate or a pharmaceutically acceptable salt thereof. nothing.

[0398] Additionally, methods are provided for inducing non-therapeutic weight loss in individuals, and such individuals The law provides an effective amount of the compound described herein, or the same, to an individual in need of such treatment. The process includes at least the step of administering a pharmaceutically acceptable salt of the substance.

[0399] In these methods, the effectiveness of the compound is demonstrated, for example, by observing a significant decrease in blood glucose levels. Observe a significant increase in insulin levels and a significant decrease in HbA1c. This can be evaluated by observing a significant decrease in body weight.

[0400] Alternatively, the compounds described herein or their pharmaceutically acceptable salts may be used if necessary. It can be used to improve bone strength in individuals. In some cases, it is not necessary. Individuals that exhibit this condition have hypoossification or hypoossoid tissue formation, or have fractures, related Joint muscle strengthening surgery, prosthetic implants, dental implants, and / or spinal fusion surgery It is healing. The polypeptides described herein can also be used for other disorders, such as Parkinson's disease. It may be used to treat Sonnson's disease or Alzheimer's disease.

[0401] In addition, the compounds described herein, or their pharmaceutically acceptable substances, for use in the treatment of therapies. A suitable salt is provided. In some embodiments, as specified herein, obesity, chronic weight management , type 2 diabetes, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis ( NASH, metabolic dyslipidemia-associated steatohepatitis (MASH), dyslipidemia, metabolic syndrome Osteoarthritis, osteoarthritis (OA), obesity-related sleep apnea (OSA), and polycystic ovary syndrome (PCOS) Polypeptides described herein or their use for the treatment of focal syndrome (PCOS) A pharmaceutically acceptable salt is provided. Furthermore, this specification is intended to induce non-therapeutic weight loss. The use of polypeptides described in the book or their pharmaceutically acceptable salts is also provided.

[0402] Additionally, obesity, chronic weight management, type 2 diabetes, non-alcoholic fatty liver disease (NAFLD) ), non-alcoholic steatohepatitis (NASH), dyslipidemia, metabolic syndrome, Osteoarthritis (OA), obesity-related sleep apnea (OSA), and polycystic ovary syndrome (PC) The compound described herein, or the drug, in the manufacture of a pharmaceutical product for treating OS) The use of scientifically acceptable salts is provided. Also, medications to induce non-therapeutic weight loss are provided. In the manufacture of [the specified product], use of the polypeptide described herein or its pharmaceutically acceptable salt. It is also provided for use.

[0403] Polypeptides, compounds, or pharmaceutical compositions described herein are provided as part of a kit. It may be provided. In some examples, the kit provides an individual with at least one polypeptide ( And optionally, a device for administering at least one additional therapeutic agent, for example, Includes syringes, automatic injectors, or pumps. Unless otherwise defined herein, All technical and scientific terms used herein are understood by those skilled in the art to which this disclosure pertains. It has the same meaning as generally understood. Similar or equivalent to those described herein. The intended method and materials may be used in the implementation or testing of polypeptides, pharmaceutical compositions, and methods. This can be done, but preferred methods and materials are described herein.

[0404] The indefinite article "a" or "an" is used to refer to an element when the context indicates that there is only one element. Unless explicitly stated otherwise, the possibility of two or more elements being present is not ruled out. Therefore, The indefinite articles "a" or "an" usually mean "at least one".

[0405] An additional, non-limiting list of embodiments is provided below. Embodiment 1. Formula:

[0406] [ka] A compound of, or a pharmaceutically acceptable salt thereof, wherein, R1 and R2 are independent of each other.

[0407] [ka] Selected from the group consisting of, R3 and R4 independently contain C1-C5 alkyl groups, which are NH2, CH2NH2. 2, and one or more of CH2CH2NH2 are optionally substituted, * This is a connection point to the sulfur atom in the cysteine ​​residue in Fc1 or Fc2, ** This is a connection point to R5 or R6, R5 and R6 are independently selected from covalent bonds or C1-C5 alkyl groups. U comprises a tertiary amine or a 1,3,5-substituted phenyl. Z is O, NH, C1~C 30 Alkyl, (OCH2CH2) m , amino acid polymer, or a combination thereof, X contains a therapeutic polypeptide, Y contains fatty acids, m is an integer between 1 and 30. Fc1 and Fc2 are compounds containing an Fc region, or pharmaceutically acceptable salts thereof. .

[0408] Embodiment 2. The compound is

[0409] [ka] The compound according to Embodiment 1, or comprising a pharmaceutically acceptable salt thereof.

[0410] Embodiment 3. The compound is

[0411] [ka] The compound according to Embodiment 1, or comprising a pharmaceutically acceptable salt thereof.

[0412] Embodiment 4. The compound is

[0413] [ka] The compound according to Embodiment 1, or comprising a pharmaceutically acceptable salt thereof.

[0414] Embodiment 5. The compound is

[0415] [ka] The compound according to Embodiment 1, or comprising a pharmaceutically acceptable salt thereof.

[0416] Embodiment 6. Polypeptide is X1X2X3X4TX6TSDX 10 X 11 X 12 X 13 LX 15 X 16 KAQX 20 X 21 FIX 24 X 25 LX 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 X 40 X 41 , comprising X1 is H, NMeY or Y, X2 is Ac4c, Aib, αMeS, Iva or D-Ala, X3 is Q, E or H, X4 is G or D-Ala, X6 is αMeF(2F), F or αMeF, X 10 is 4-Pal, Y or V, X 11 is Aib, S or αMeS, X 12 is I or S, X 13 is L or αMeL, X 15 is D or E, X 16 is E or Orn, X 20 is Aib, Iva or αMe4Pal, X 21 is E, Q, D or Orn X 24 is K, Q, E, D-Glu or D-Gln, X 25 is αMeY or Y, X 27 is L or V, X 28 is K, E or A, X 29 is Aib or G, X 30 is G or S, X 31 is P, G, E or Orn X 32 is absent, or S or P, X 32 is S or P, X 33 is S, X33 If X is S, 34 is G or Aib, X 34 If X is G or Aib, 35 It does not exist, or it is A or Orn. the law of nature, X 35 If X is A or Orn, 36 It does not exist, or it is P. X 36 If P, then X 37 It does not exist, or it is P. X 37 If P, then X 38 It does not exist, or it is P. X 38 If P, then X 39 It does not exist, or it is S, Orn, or G. X 39 If is S, Orn, or G, then X 40 It does not exist, or it is K or G. X 40 If it is K or G, then X 41 It does not exist, or it is S or G. X 32 If X does not exist, 33 ~X 41 It does not exist, X 35 If X does not exist, 36 ~X 41 It does not exist, X 36 If X does not exist, 37 ~X 41 It does not exist, X 37 If X does not exist, 38 ~X 41 It does not exist, X 38 If X does not exist, 39 ~X 41 It does not exist, X 39 If X does not exist, 40 and X 41 It does not exist, X 40If X does not exist, 41 It does not exist. The C-terminal amino acid is optionally amidated, or is a pharmaceutically acceptable salt thereof. A compound described in any one of the application methods 1 to 5.

[0417] Embodiment 7.Z is one of the positions 24, 28, 31, or 40 of the polypeptide The compound according to Embodiment 6, which is conjugated to an amino acid.

[0418] Embodiment 8.Z is a compound comprising an amino acid polymer as described in any one of Embodiments 1 to 7. Compounds, or pharmaceutically acceptable salts thereof.

[0419] Embodiment 9. The amino acid polymer is (GGGGS)n, (SGGGG)n, (GGGG Q)n, (EAAAK)n, (KAAAE)n, (AEEA)n, G(PA)n, G(P A)n, G(PE)n, G(PK)n, (AP)n, (AP)n, or G(EP)n A compound according to Embodiment 8, which contains any of the above, where n is an integer from 1 to 10, or the same Pharmaceutically acceptable salts.

[0420] Embodiment 10. Lysine at position 17 is an amino acid and C 16 ~C 22 Linker between fatty acids via C 16 ~C 22 Conjugated to a fatty acid, in any of Embodiments 1 to 9 The compound described, or a pharmaceutically acceptable salt thereof.

[0421] Embodiment 11.C 16 ~C 22 The fatty acid is conjugated to lysine at position 17 via the linker. A gated compound, or a pharmaceutically acceptable salt thereof, as described in Embodiment 10.

[0422] Embodiment 12. The compound according to Embodiment 11, wherein the linker contains 1 to 4 amino acids. or a pharmaceutically acceptable salt thereof.

[0423] Embodiment 13. The amino acids contained in the linker are Glu, γGlu, or a combination thereof. A compound according to Embodiment 12, or a pharmaceutically acceptable salt thereof.

[0424] Embodiment 14. The linker has 1 to 4 (2-[2-(2-amino-ethoxy)-ethoxy A compound according to any of Embodiments 11 to 13, including the xy-acetyl moiety, or the A pharmaceutically acceptable salt of [the substance].

[0425] Embodiment 15. The linker is (γGlu) a -(2-[2-(2-amino-ethoxy) (-ethoxy]-acetyl) b -(γGlu) c -CO-(CH2) p -The structure of CO2H Including, in the formula, a is 0 or 1, b is 0, 1, or 2, c is 1, 2, or 3, p is an integer between 14 and 20, the compound described in Embodiment 14, or its pharmaceutically acceptable Salt.

[0426] Embodiment 16.a is 0, b is 1, c is 1, and the chemical formula described in Embodiment 15 Compounds, or pharmaceutically acceptable salts thereof.

[0427] Embodiment 17. A polypeptide is found to be in a ratio of more than 90% with any of SEQ ID NOs: 1 to 482. A compound according to any one of Embodiments 1 to 16, including serial identity, or a pharmaceutically acceptable compound thereof. Salt is acceptable.

[0428] Embodiment 18. The polypeptide comprises one of the sequences from SEQ ID NOs: 1 to 482. The compound described in Embodiment 17, or a pharmaceutically acceptable salt thereof.

[0429] Embodiment 19. Fc1 and Fc2 are, a.AGCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVV VDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVV SVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQP REPQVYTLPSQEEMTKNQVSLTCLVKGFYPSDIAVEWES NGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVF SCSVMHEALHNHYTQKSLSLSLG (Sequence ID 935), b.AGCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVV VDVSQEDPEVKFNWYVDGVEVHNAKTKPREEQFNSTYRVV SVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQP REPQVYTLPSREEMTKNQVSLTCLVKGFYPSDIAVEWES NGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSLG (Sequence ID 936), c.IGCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVV VDVSQEDPEVKFNWYVDGVEVHNAKTKPREEQFNSTYRVV SVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQP REPQVYTLPSREEMTKNQVSLTCLVKGFYPSDIAVEWES NGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSLG (Sequence ID 941), d.IGCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVV VDVSQEDPEVKFNWYVDGVEVHNAKTKPREEQFNSTYRVV SVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQP REPQVYTLPSREEMTKNQVSLTCLVKGFYPSDIAVEWES NGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSLG (Sequence ID 941), e.AGCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVV VDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV SVLTVLHQDWLNGKEYKCKVSNKALAAPIEKTISKAKGQP REPQVYTLPSREEMTKNQVSLTCLVKGFYPSDIAVEWES NGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGK (Sequence ID 943), f.AGCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVV VDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQP REPQVYTLPSREEMTKNQVSLTCLVKGFYPSDIAVEWES NGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGK (Sequence ID 944), g.ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSQEDPEVKFNWYVDGVEVHNAKTKPREEQ FNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKT ISKAKGQPREPQVYTLPSREECTKNQVSLTCLVKGFYPS DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKS RWQQGNVFSCSVMHEALHNHYTQKSLSLSLG(Sequence ID 1179 ), h.ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSQEDPEVKFNWYVDGVEVHNAKTKPREEQ FNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKT ISKAKGQPREPQVYTLPSREEMTKNQVSLTCLVKGFYPS DIAVEWESNGQPENNYKTTPPVCDSDGSFFLYSRLTVDKS RWQQGNVFSCSVMHEALHNHYTQKSLSLSLG(Sequence ID 1180 ), i.ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSQEDPEVKFNWYVDGVEVHNAKTKPREEQ FNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKT ISKAKGQPREPQVYTLPSREEMTKNQVSLTCLVKGFYPS DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKC RWQQGNVFSCSVMHEALHNHYTQKSLSLSLG(Sequence ID 1181) ) j.ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISR TPEVTCVVVDVSQEDPEVKFNWYVDGVEVHNAKTKPREEQ FNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKT ISKAKGQPREPQVYTLPSREEMTKNQVSLTCLVKGFYPS DIAVEWESNGQPENNYKTTPPVLDSDGSFLLYSKLTVDKS RWQQGNVFSCSVMHEALHNHYTQKSLSLSLG(Sequence ID 948) A compound selected from the group consisting of, any one of embodiments 1 to 18, or the compound described therein Pharmaceutically acceptable salts.

[0430] Embodiment 20. The pharmaceutically acceptable salt is sodium, potassium, trifluoroacetate A compound selected from hydrochloride and acetate, as described in any one of Embodiments 1 to 19. , or a pharmaceutically acceptable salt thereof.

[0431] Embodiment 21. A compound comprising any of the following Fc-acylated polypeptides.

[0432] [Table 13-1]

[0433] [Table 13-2]

[0434] [Table 13-3]

[0435] Table 13-4

[0436] Table 13-5

[0437] Table 13-6

[0438] Table 13-7

[0439] Table 13-8

[0440] Table 13-9

[0441] Table 13-10

[0442] Table 13-11

[0443] Table 13-12

[0444] Table 13-13

[0445] Table 13-14

[0446] Table 13-15

[0447] Table 13-16

[0448] Table 13-17

[0449] Table 13-18

[0450] Table 13-19

[0451] Table 13-20

[0452] Table 13-21

[0453] Table 13-22

[0454] Table 13-23

[0455] [Table 13-24]

[0456] [Table 13-25]

[0457] [Table 13-26]

[0458] [Table 13-27]

[0459] [Table 13-28]

[0460] [Table 13-29]

[0461] [Table 13-30]

[0462] [Table 13-31]

[0463] Embodiment 22. A compound comprising any of the following Fc-acylated polypeptides.

[0464] [ka]

[0465] [ka]

[0466]

change

[0467]

change

[0468]

change

[0469]

change

[0470]

change

[0471]

change

[0472]

change

[0473]

change

[0474]

change

[0475]

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[0476]

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[0490] Embodiment 22. The compound is natural GIP (SEQ ID NO: 1223) and GLP-17-36 ( Compared to SEQ ID NO: 1221, the results for both GIP and GLP-1 receptors are larger. A compound described in any one of Embodiments 1 to 22 that has strong efficacy, or a compound thereof pharmaceutically Allowable salt.

[0491] Embodiment 23. The compound is natural glucagon (SEQ ID NO: 1222), GIP (SEQ ID NO: 1222). 223) and GLP-1 7-36 Compared to (Sequence ID 1221), glucagon, GIP The embodiments 1 to 22, which have greater efficacy in each of the GLP-1 receptors. Any one of the compounds listed, or a pharmaceutically acceptable salt thereof.

[0492] Embodiment 24. The compound is sufficiently extended to enable low-frequency dosing, such as once a month. A compound according to any one of Embodiments 1 to 23, or a drug having a duration of action. A scientifically acceptable salt.

[0493] Embodiment 25. The compound described in any one of Embodiments 1 to 24 or a pharmaceutically acceptable compound thereof. A salt comprising, and at least one pharmaceutically acceptable carrier, diluent, or excipient, Pharmaceutical composition.

[0494] Embodiment 26. The composition described in Embodiment 25, wherein the composition is formulated for subcutaneous administration. Pharmaceutical composition.

[0495] Embodiment 27. Diabetes, obesity, chronic weight management, non-alcoholic fatty liver disease (NAFLD) ), non-alcoholic steatohepatitis (NASH), dyslipidemia, metabolic syndrome, Chronic kidney disease (CKD), osteoarthritis (OA), obesity-related sleep apnea (OSA), polycystic sacral ventricles Select from the group consisting of follicular ovary syndrome (PCOS), Parkinson's disease, and Alzheimer's disease. A method for treating a selected disease or condition, which involves administering an effective amount to an individual in need. Administer a compound described in any one of Forms 1 to 26, or a pharmaceutically acceptable salt thereof. A method that includes a process.

[0496] The present invention will be further described by the following embodiments, but these should be construed as limiting. It shouldn't be done. [Examples]

[0497] Preparation of therapeutic polypeptides The processes for producing the exemplary compounds of this disclosure are described below. These processes are extremely efficient. Rather than being prepared using other processes known to those skilled in the art, the compounds of this disclosure can be prepared using other processes. It will become clear that this is also possible.

[0498] Peptide intermediates intended for Fc conjugation are conventional fluorenylmethyl oxy Sicarbonyl (Fmoc) / tert-butyl (t-Bu) solid-phase peptide synthesis (SPPS) The entire process is synthesized according to the protocol. This process is called Symphony 12-C. hannel Multiplex Peptide Synthesizer(Pro This was carried out using Tein Technologies, Inc. (Tucson, AZ). It can be done.

[0499] A lysine residue (4-methyltrityl (Mtt)) is orthogonally protected at the specified sequence position. First, a linear peptide skeleton having (and ivDde) ​​is synthesized. This is followed by a continuous The deprotection and coupling process separates the "linker-fatty acid" portion, followed by the "linker-F" portion. The "c conjugation" part is introduced. These modifications result in the following: Then, the synthesis of the peptide intermediates is completed, they are cleaved from the solid support, and then purified. ru.

[0500] Procedure for peptide synthesis Solid-phase peptide synthesis (SPPS) is performed in a substitution range of 0.3–0.6 mmol / g with 1% DV B Cross-linked polystyrene resin (Fmoc-Rink-MBHA, 100-200 mesh, C This is done using hem-Impex International. The peptide backbone is, The residue at position 1 (for example, Boc-L-Tyr(OtBu)-OH or Boc-L-His( Boc)-OH), lysine residue at position 17 (Fmoc-L-Lys(Mtt)-OH), The lysine residue at positions 24, 28, 32, or 40 (Fmoc-L-Lys(ivDde) Assembled with N-Fmoc protected amino acids containing standard side-chain protecting groups, except for the -OH group. Before each coupling step, the Fmoc group is introduced using 20% ​​piperidine in DMF. Remove (2 / 8 treatments each). To initiate coupling, equimolar amounts of Fm oc amino acids (0.3M in DMF), diisopropylcarbodiimide (0.9M in DCM) ), and Oxyma (0.9M in DMF) are added in a 9-fold molar excess relative to the theoretical peptide packing. Then, the reaction is carried out while maintaining the temperature at 60°C. Coupling times vary, and standard F moc-amino acids are coupled for 40 minutes, but Fmoc-α-methylated amino acids This process takes 3 hours, similar to how Fmoc-amino acids are coupled to α-methylated residues. The extended coupling time of 6 hours is due to the Fmoc-amino acids at positions 5 and 6. The peptide skeleton synthesis is performed, while the residues at positions 1-4 undergo coupling for 3 hours. After completion, the resin is thoroughly cleaned with DCM to remove any remaining DMF.

[0501] Linker - Procedure for assembling the fatty acid portion After peptide skeleton synthesis, the Mtt protecting group on the lysine at position 17 is removed using 30% hexaphosphate in DCM. Treatment with ruoroisopropanol (four 30-minute treatments), followed by thorough treatment with DCM. Selectively removed by washing. The linker-fatty acid portion is removed as needed by selecting N- Using an Fmoc-protected amino acid derivative, the same coupling approach as in the previous step was used. These are assembled as follows: 2-[2-(2-Fmoc-amino-ethoxy)-ethoxy [Fmoc-AEEA-OH]-acetic acid, Fmoc-α-t-butyl ester glutamate Ter (Fmoc-Glu(OH)-OtBu), Nα-tert-butoxycarbonyl- Nε-Fmoc-lysine (Boc-L-Lys(Fmoc)-OH), and 18-(te It may contain rt-butoxy)-18-oxo-octadecanoic acid. Each derivative may contain diisopropyl Pyrcarbodiimide (9 equivalents, 0.9M in DCM) and Oxyma (9 equivalents, 0 in DMF) In the presence of 0.9M, 4 to 9 equivalents of resin filling (0.15 to 0.3M in DMF) Coupling occurs according to the amount in each cup. The coupling reaction is carried out at 60°C for 6 hours. Before the ringing process, the Fmoc group is removed using 20% ​​piperidine in DMF (each (Processing 2 out of 8 times).

[0502] Procedure for assembling the linker-Fc conjugation section After assembling the linker-fatty acid moiety, on the lysine residue at position 24, 28, 32, or 40 The ivDde protecting group is treated with 3% hydrazine in DMF (each treatment being 4 / 3 of a treatment). The resin is selectively removed by DMF, DCM, and isopropyl alcohol. Wash thoroughly to completely remove any residual hydrazine. If a linker is present, Using an N-Fmoc-protected amino acid derivative selected as needed, the previous coupling These are constructed using a similar approach to the process. These are Fmoc-N-amide-PEG. 24 -Acid, 2-[2-(2-Fmoc-amino-ethoxy)-ethoxy]-acetic acid (Fmo It may contain c-AEEA-OH, or Fmoc-amino acids having a standard protecting group. The derivatives are diisopropylcarbodiimide (9 equivalents, 0.9 M in DCM) and Oxyma In the presence of (9 equivalents, 0.9 M in DMF), resin filling (0.15~0.3 M in DMF) In contrast, coupling occurs in amounts ranging from 2 to 9 equivalents. The coupling reaction is carried out as needed. The process is carried out at 60°C for 2 to 6 hours. Before each coupling step, the Fmoc group is added to the DMF. Remove using 20% ​​piperidine (two 8-minute treatments each).

[0503] BEA coupling After the desired linker is assembled, the terminal Fmoc group is added to DMF with 20% piperidine. Remove using (2 / 8 treatments each). Then, 4-(screw(2-((((9H-full Oren-9-yl)methoxy)carbonyl)amino)ethyl)amino)-4-oxobuta Acid (Fmoc2-BEA-OH, 6 equivalents in DMF), diisopropylcarbodiimide ( Add 9 equivalents (0.9 M in DCM) and Oxyma (9 equivalents (0.9 M in DMF)). This is used to perform coupling to "BEA". The reaction is maintained at 60°C for 4 hours. Once completed, lead the desired thiol-reactive conjugation portion as detailed below. Before insertion, remove both Fmoc groups with 20% piperidine in DMF (2 / 8 of each). process).

[0504] In the case of a peptide intermediate containing BEA-(MalDap)2 (S)-3-((tert-butoxycarbonyl)amino)-2 is added to the resin-bonded diamine. -(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl)propanoic acid (N -Mal-L-Dap(Boc)-OH (3.6 equivalents in DMF), diisopropylcarb Diimide (9 equivalents, 0.9M in DCM), and Oxyma (9 equivalents, 0.9M in DMF) Introduce the solution and allow the reaction to proceed at room temperature for 3 hours.

[0505] In the case of a peptide intermediate containing BEA-(MSPT)2 The resin-bound diamine contains N,N-diisopropylethylamine (1.7 equivalents) and 2,5 -Dioxopyrrolidine-1-yl2-(2-(2-(4-(5-(methylsulfonyl)- 1H-Tetrazole-1-yl Phenoxy Ethoxy Ethoxy Acetate (MSP) Add T-NHS ester (3 equivalents in DMF) and allow the reaction to proceed at room temperature for 2.5 hours.

[0506] In the case of a peptide intermediate containing BEA-(AcBr)2 N,N-diisopropylethylamine (1.7 equivalents) and 2,5-dioxopyrrolidine -1-yl-2-bromoacetate (4.1 equivalents in DMF) is added to the resin-bound diamine. Allow the reaction to proceed at room temperature for 2.5 hours.

[0507] In the case of a peptide intermediate containing BEA-(OD)2 N,N-diisopropylethylamine (1.7 equivalents) and 2,5-dioxopyrrolidine -1-yl2-(2-(2-(4-(5-(methylsulfonyl)-1,3,4-oxadi Azole-2-yl Phenoxy Ethoxy Ethoxy Acetate (OD-NHS S) Add tel (3 equivalents in DMF) to the resin-bonded diamine and allow the reaction to proceed at room temperature for 2.5 hours. ru.

[0508] In the case of a peptide intermediate containing BEA-(β-MalDap)2 The resin-bound diamine contains N,N-diisopropylethylamine (1.7 equivalents) and 2,5 -Dioxopyrrolidine-1-yl(S)-2-((tert-butoxycarbonyl)amine (no)-3-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1-yl)propane Add noate (Boc-L-Dap(Mal)-NHS ester, 3 equivalents in DMF) The mixture was allowed to progress overnight at room temperature.

[0509] In the case of a peptide intermediate containing a single MalDap (conjugate to Fc-eCys) (For the purpose of) (S)-3-((tert-butoxyca Rubonyl)amino)-2-(2,5-dioxo-2,5-dihydro-1H-pyrrole-1 -yl)propanoic acid (N-Mal-L-Dap(Boc)-OH, 1.8 equivalents in DMF) , diisopropylcarbodiimide (9 equivalents, 0.9M in DCM), and Oxyma (9 equivalents Add 0.9 M of DMF and allow the reaction to proceed at room temperature for 3 hours.

[0510] In the case of a peptide intermediate containing a single MSPT (conjugation to Fc-eCys) (For the sake of) Resin-bound amine (BEA is not present), 2-(2-(2-(4-(5-(methylsulfide Rufonyl)-1H-tetrazol-1-yl)phenoxy)ethoxy)ethoxy)acetic acid MSPT-acid (1.8 equivalents in DMF), diisopropylcarbodiimide (9 equivalents, DCM) Add 0.9M of ethanol and Oxyma (9 equivalents, 0.9M in DMF), and carry out the reaction at room temperature for 3 minutes. Advance time.

[0511] Cleavage and purification of peptide intermediates After the resin synthesis process described in detail above is completed, the peptide resin is washed with DCM, and then , allow to air dry completely. Cut the resin into 10 mL of the cutting cocktail (trifluoroacetic acid:water:tri Treat with isopropylsilane (85:5:10 v / v) at room temperature for 2.5 hours. Filter the resin. Separate the samples, wash each twice with 2 mL of undiluted TFA, and combine the filtrates. Dilute the filtrate five times (by volume). Treat with cold diethyl ether (-20°C) to precipitate the crude peptide. Then, The butyl / ether suspension is centrifuged at 5500 rpm for 2 minutes to form a solid pellet. The supernatant is drained, the solid pellets are further ground twice with ether, and dried under nitrogen. Crude Peptide Dissolve the cydorhizon in 20% acetonitrile / 20% acetic acid / 60% water, and buffer with 0.1% TFA / water. Linear gradient of 100% acetonitrile in a liquid system (e.g., 30-50% acetonitrile over 75 minutes) Luna 5μm phenylhexyl preparative column (21×250mm, P) using tonitrile The product is purified by RP-HPLC using henomenex. The purity of the product is measured using analytical RP-HPLC. Evaluated using PLC, the pooling criterion is over 95%. The pooled fractions are frozen. The peptide intermediate is concentrated by freeze-drying to obtain a TFA salt.

[0512] 4-(bis(2-((((9H-fluoren-9-yl)methoxy)carbonyl)amide Synthesis of ethylamino-4-oxobutanoic acid (Fmoc2-BEA-OH)

[0513] [ka] N-(9-Fluorenylmethoxycarbonyloxy) in dichloromethane (80 mL) A solution of succinimide (19.4 g, 56.4 mmol) was cooled to -78°C and mixed with dichloroethylene. In a solution of diethylenetriamine (3.1 mL, 28 mmol) in lomethane (30 mL) The mixture was added over 45 minutes. After 2 hours, the mixture was warmed to ambient temperature, and succinic anhydride (9.91%) was added. (g, 98 mmol) and DMAP (691 mg, 5.60 mmol) were added. Stir the mixture at ambient temperature for 15 hours, then slowly add 1N HCl (approximately 20 mL). The pH was adjusted to 5. The phases were separated, and the aqueous layer was extracted using DCM (2 × 100 mL). The combined organic layers were washed with saturated NaCl aqueous solution and then dried with magnesium sulfate. The solvent was removed under reduced pressure, and silica gel chromatography (660g, DCM(5)) was performed. (minutes), then 5% MeOH / DCM (15 minutes), then 10% MeOH / DCM (25 minutes) The product was purified to obtain the title product (911.84 g, 65%) as a white powder. ES / MS m / z:648(M+1).

[0514] 2,5-Dioxopyrrolidine-1-yl 2-(2-(2-(4-(5-(methylsulfon Nyl)-1H-tetrazol-1-yl)phenoxy)ethoxy)ethoxy)acetate Synthesis of (MSPT-NHS ester) Process 1

[0515] [ka] 4-(5-mercapto-1H-tetrazole-1-yl)phenol (4.00g, 2 Dissolve 0.6 mmol) in tetrahydrofuran (50 mL), and cool the mixture in an ice bath. Then, N,N-diisopropylethylamine (4.31 g, 33.3 mmol) was added. After stirring for 10 minutes, a suspension was formed. Iodomethane (1.54 mL, 24.7 mmol was added dropwise through a syringe over 1 minute. The reaction mixture was cooled in an ice bath. The mixture was stirred for 20 minutes, then at room temperature for 12 hours. The mixture was diluted with acetaldehyde (100 mL). The mixture was washed with saturated NH4Cl aqueous solution (2 × 50 mL). The organic layer was separated, and then sodium sulfate was added. Dry with alium, filter, and concentrate in vacuum to obtain 4-(5-(methylthio)-1H-teto Razole-1-yl)phenol (4.2g, 93% yield) was obtained. This was further purified. It can be used directly in the next process without any issues. LCMS-mz=209(M+1).

[0516] Process 2

[0517] [ka] In a 200 mL pressure vessel, add 4-(5-(methylthio)-1H-tetrazole-1-yl )Phenol (2.50g, 11.4mmol), tert-butyl 2-(2-(2-butyl Romoethoxy(ethoxy)acetate (4.33g, 14.8mmol) and acetone ( I added 60 mL. I added potassium carbonate (3.15 g, 22.8 mmol) and closed the container. The mixture was sealed and heated at 80°C for 8 hours with vigorous stirring. The reaction mixture was cooled to room temperature and filtered. Remove potassium carbonate, then wash with acetone / DCM / siRNA (30 mL each). The filtrate was concentrated in a vacuum to obtain the crude substance, which was then subjected to flash chromatography (8 (0g, 100% DCM for 5 minutes, then a gradient to 100% siRNA over 20 minutes) The product was purified. The product is tert-butyl 2-(2-(2-(4-(5-(methylthio)-1 H-tetrazole-1-yl phenoxy ethoxy ethoxy acetate (3.98 g, yield 85%) was isolated as a white powder. LCMS-mz=411(M+1).

[0518] Process 3

[0519] [ka] tert-butyl 2-(2-(2-(4-(5-(methylthio)-1H-tetrazol -1-yl(phenoxy(ethoxy)(ethoxy)(ethoxy)acetate (3.98g, 9.21mm) Dissolve ol) in ethanol (100 mL), cool in an ice bath to 5-10°C, then 30 % hydrogen peroxide aqueous solution (19.0 mL, 184 mmol), followed by molybdic acid (VI) Ammonia tetrahydrate (1.14 g, 0.921 mmol) was added. The reaction mixture was prepared at room temperature. The mixture was stirred in an ice bath for 4 hours, then stirred at room temperature for 12 hours. The mixture was then mixed in DCM (150 mL). Diluted with ) and then washed with brine. The organic phase was separated and dried with sodium sulfate. Concentrated to dryness in a vacuum. Flash column chromatography (80g silica, 100%). Purify using a gradient to 100% siRNA for 5 minutes in DCM, followed by 20 minutes. -(2-(2-(4-(5-(methylsulfonyl)-1H-tetrazole-1-yl) Ethoxy(ethoxy)(ethoxy)acetic acid (MSPT acid, 3.00g, yield 80%) concentrated It was obtained as an oily substance. LCMS-mz=385(M-1)

[0520] Process 4

[0521] [ka] 1-Hydroxypyrrolidine-2,5-dione (1.33g, 1.6 equivalents, 11.6mm) ol) in DCM (50 mL) and THF (70 mL) 2-(2-(2-(4-(5 -(methylsulfonyl)-1H-tetrazole-1-yl)phenoxy)ethoxy)eth It was added to a solution of xy)acetic acid (2.80 g, 7.25 mmol). EDCI (1.60 g) (10.3 mmol) was added all at once, and at that point the solution became a cloudy mixture. Add 20 mL of DCM to the mixture to restore it to a solution, and then stir at room temperature for 12 hours. The solvent was removed in a vacuum, and the crude substance was obtained as a white foam. Flash column chromatography Graph (80g, 100% DCM for 5 minutes, followed by 100% toluene for 20 minutes) Purification by gradient to 2,5-dioxopyrrolidine-1-yl2-(2-(2 -(4-(5-(methylsulfonyl)-1H-tetrazole-1-yl)phenoxy) Toxy(ethoxy)acetate (MSPT-NHS ester, 2.61g, yield 65%) It was obtained as a low-melting-point solid. LCMS-mz=484(M+1)

[0522] 2,5-Dioxopyrrolidine-1-yl 2-(2-(2-(4-(5-(methylsulfon Nyl)-1,3,4-Oxadiazole-2-yl)phenoxy)ethoxy)ethoxy) Synthesis of acetate (OD-NHS ester) Process 1

[0523] [ka] 4-(5-mercapto-1,3,4-oxadiazole-2-yl)phenol (3. (00g, 15.4 mmol) was dissolved in THF (50mL) and cooled to 0°C in an ice bath. N,N-diisopropylethylamine (3.46 mL, 2.60 g, 20.1 mmol) ) was added to obtain a turbid solution. The mixture was stirred in an ice bath for 5 minutes, and then iodomethane ( 2.85 g (20.1 mmol) was added dropwise through a syringe over 1 minute. The mixture became clear after 5 minutes. Remove from the cooling bath and stir the mixture at room temperature for 2 hours. Then, it was diluted with dichloromethane (100 mL) and washed with saturated NH4Cl aqueous solution ( The pH was adjusted to approximately 5 by adding 2 x 50 mL of enoic acid solution. Separate, dry with sodium sulfate, concentrate to dryness in vacuum, and obtain 4-(5-methylsulfate Nyl-1,3,4-oxadiazole-2-yl)phenol (520 mg, yield 97%) ) was obtained as a pale yellow solid and used in the next step without further purification. LC-MS- mz = 209(M+1).

[0524] Process 2

[0525] [ka] 4-(5-(methylthio)-1,3,4-oxadiazole-2-yl)phenol ( Add 3.3g (1 equivalent, 15 mmol) and acetone (60 mL) to a 200 mL pressure vessel. Added to this solution: tert-butyl 2-(2-(2-bromoethoxy)ethoxy) Acetate (5.5g, 20 mmol) and potassium carbonate (4.2g, 30 mmol) The substance was added. The pressure vessel was sealed and heated at 70°C for 5 hours with vigorous stirring. It was then cooled to room temperature. Afterward, the mixture was filtered to remove solid potassium carbonate and washed with siRNA / DCM. The filtrate was concentrated to dryness under vacuum, and normal-phase flash column chromatography was performed (80g silica gel). (Gradient, 100% DCM for 5 minutes, then a gradient to 100% siRNA over 20 minutes) Further purification was performed. The product-containing fraction was concentrated in vacuum and tert-butyl 2-(2-(2- (4-(5-(methylthio)-1,3,4-oxadiazole-2-yl)phenoxy) Ethoxy(ethoxy)acetate (4.8g, 74% yield) was obtained as a white solid. LC -MS-mz=411(M+1).

[0526] Process 3

[0527] [ka] tert-butyl2-(2-(2-(4-(5-(methylthio)-1,3,4-oxa Diazole-2-yl phenoxyethoxyethoxyacetate (5.20g, 1 Dissolve 2.7 mmol) in 100 mL of ethanol and cool to 5-10°C in an ice bath. Next, 30% hydrogen peroxide (10 mL, 97 mmol), followed by ammonium molybdate (VI) 501 mg of nium tetrahydrate (0.405 mmol) was added. The mixture was vigorously stirred for 2 hours. Then, an additional 15 mL of 30% hydrogen peroxide and 1 g of ammonium(VI) molybdate IV Hydrate was added. The reaction mixture was stirred for a further 6 hours, and then diluted with 150 mL of DCM. The mixture was washed with brine. The organic phase was separated, dried with sodium sulfate, and concentrated to dryness in a vacuum. The residue was then pulverized with methanol to obtain the first part of the product. The solvent was then reduced under reduced pressure. Removed from the mother liquor below. Flash column chromatography (40g, 100% DCM) Purification by a gradient (to 100% siRNA) for 3 minutes, then 20 minutes, is performed. The product was obtained as a white solid. Combining both product moieties, tert-butyl 2-( 2-(2-(4-(5-(methylsulfonyl)-1,3,4-oxadiazole-2-i Phenoxyethanol acetate (5.3g, 90% yield) is a white solid. The result was obtained as follows: LC-MS-mz = 387(M-tBu).

[0528] Process 4

[0529] [ka] tert-butyl 2-(2-(2-(4-(5-(methylsulfur in DCM (60 mL)) Honyl)-1,3,4-Oxadiazole-2-yl)phenoxy)ethoxy)ethoxy ) In a solution of acetate (5.60 g, 12.0 mmol), add 2,2,2-trifluorovinegar Acid (20 mL, 12.0 mmol) was added. The reaction mixture was stirred at room temperature for 2 hours, and then... The solution is then concentrated under vacuum to obtain a concentrated residue, which is then subjected to normal-phase flash column chromatography. (80g silica gold column, 100% DCM for 3 minutes, then 20 minutes for 10 The product was purified by a gradient to 0% siRNA. The product-containing fractions were combined and concentrated in a vacuum. , 2-(2-(2-(4-(5-(methylsulfonyl)-1,3,4-oxadiazole -2-yl)phenoxy)ethoxy)ethoxy)acetic acid (4.12 g, yield 82%) was obtained. LCMS-mz=387(M+1).

[0530] Process 5

[0531] [ka] 2-(2-(2-(4-(5-(methylsulfonyl)-1,3,4-oxadiazole -2-yl(phenoxy(ethoxy)(ethoxy)acetic acid (3.00g, 7.38mmol) and 1-hydroxypyrrolidine-2,5-dione (1.19 g, 10.3 mmol) D It was dissolved in CM (50 mL) and THF (70 mL). To this solution, 3-(((ethyl ethyl Mino(methylene)amino)-N,N-dimethylpropane-1-amine(EDCI, 1.6 0g (10.3 mmol) was added. Upon addition, the solution became cloudy, and additional DCM (20m) was added. L) was added to the mixture to restore it to a solution, and then the mixture was stirred at room temperature for 12 hours. The solution was concentrated under reduced pressure, the resulting residue was dissolved in DCM, and normal-phase flash column chromatography was performed. Fee (80g Silica Gold, 100% DCM for 5 minutes, then 100% for 20 minutes) The product was purified by a gradient to siRNA. The product-containing fraction was concentrated to extract 2,5-dioxopy Roridine-1-yl-2-(2-(2-(4-(5-(methylsulfonyl)-1,3,4 -Oxadiazole-2-yl)phenoxy)ethoxy)ethoxy)acetate (2.6 A 1g sample was obtained with a yield of 65%. LCMS-mz = 484(M+1).

[0532] Fc recrosslinking Fc is obtained by adding 1.5 to 2 equivalents of newly prepared TCEP stock aqueous solution. This reduces the solution. 258 μL of 7 mM TCEP solution is added to 9 mL of 111 μM Fc(pH) Add to 7.5). Then, incubate the solution at 37°C for 1 hour and analyze by LC-MS. The reduction is confirmed by analysis. Next, the reduced Fc is conjugated with the peptide. Use directly for conjugation. For efficient conjugation, the reaction is appropriate. Using an excess amount (approximately 1.4 equivalents) of peptide, at an Fc concentration of 21.2 μM, the following was performed: The procedure is carried out as follows: Transfer the reduced Fc (approximately 9 mL) to a 50 mL vial, and add 50 mM to it. Add approximately 29 mL of acetate buffer (pH 5.6) and 6 mL of ACN. Gently stir the solution. After mixing and standing at 4°C, add a 0.5 mM stock solution in 30% acetonitrile water. Then the peptide is added. The progress of the reaction is monitored by LC-MS, and typically 30 Completed within minutes. To promote the hydrolysis of the maleimide ring, use 1M tonic water at pH 8.0. The reaction mixture is adjusted to pH 8.0 by adding buffer solution. The desired product is then obtained from I It is isolated after purification using EX.

[0533] eCys conjugation for alternative peptide binding sites First, add 8-10 molar equivalents of dithiothre to Fc, which has been properly manipulated to contain cysteine. The sample was reduced at 37°C for 2 hours in the presence of yitol (DTT). After reduction, the sample was treated with Zeba Spin desalination column (Thermo Fisher Scientific) or alternatively The cells were subjected to size exclusion chromatography to bind the reducing agent and the cysteine ​​manipulated during expression. The combined cysteine ​​or glutathione cap was removed. Then, 1 to 5 molar equivalents of decyl phosphate were added. Add droascorbic acid (DHAA) and maintain this sample at room temperature for 0.5 to 1 hour. Then, the hinged disulfide was reformed. After oxidation, the sample was again subjected to Zeba spin column ( (Surface Fisher Scientific), or alternatively, size exclusion chromatographs The sample is passed through a tography column to remove the oxidizing agent and then used for conjugation. The buffer solution was replaced with the desired buffer solution.

[0534] eCys conjugation for single peptides To generate an Fc having a single eCys site, a 1:1 ratio of Fcs having the desired site is used. The mixture was prepared in the presence of 8-10 molar equivalents of dithiothreitol (DTT) in F405L and Fc containing the R409K mutation was mixed and incubated at 37°C for 2 hours. After reduction, The agent and the cysteine ​​or glutathione caps bound to the cysteine ​​that were manipulated during expression Then, it was removed using either a desalting column or size exclusion chromatography. Add 1-5 molar equivalents of dehydroascorbic acid (DHAA) and let the sample cool to room temperature. Maintain for 0.5 to 1 hour to change the F405L and R409K in one Fc molecule in the mixture. Hinged disulfide was reformed preferentially by hetero-driven hetero-Fc formation. Next, the sample was placed back into the Zeba spin column (Thermo Fisher Scient). (Issue) or, alternatively, pass through a size exclusion chromatography column to remove the oxidizing agent. The sample was then buffer-changed to the desired buffer for conjugation.

[0535] Conjugation to single or dual eCys Fc constructs For efficient conjugation to the eCys variant, the reaction is carried out in appropriate excess amounts. (Approximately 1.4 equivalents for single eCys, approximately 2.8 equivalents for double eCys) Using this method, the procedure is carried out at an Fc concentration of 21.2 μM as follows: Fc (approximately 9 mL) is used in 5 Transfer to a 0 mL vial and add 50 mM acetate buffer (approximately 29 mL) pH 5.6 and 6 mL. Add L of ACN. Gently mix the solution and let it stand at 4°C, then add 30% acetonite in water. The peptide is added as a 0.5 mM stock solution in Toll. The reaction is monitored by LC-M. It is monitored by S and typically completed within 30 minutes. It promotes hydrolysis of the maleimide ring. To proceed, add 1M Tris buffer solution at pH 8.0 to the reactants. Adjust to 8.0. Isolate the desired product after purification by IEX.

[0536] [Table 14-1]

[0537] [Table 14-2]

[0538] [Table 14-3]

[0539] Table 14-4

[0540] Table 14-5

[0541] Table 14-6

[0542] Table 14-7

[0543] Table 14-8

[0544] Table 14-9

[0545] Table 14-10

[0546] Table 14-11

[0547] Table 14-12

[0548] Table 14-13

[0549] Table 14-14

[0550] Table 14-15

[0551] Table 14-16

[0552] Table 14-17

[0553] Table 14-18

[0554] Table 14-19

[0555] Table 14-20

[0556] Table 14-21

[0557] Table 14-22

[0558] Table 14-23

[0559] Table 14-24

[0560] Table 14-25

[0561] Table 14-26

[0562] Table 14-27

[0563] Table 14-28

[0564] Table 14-29

[0565] Table 14-30

[0566] Table 14-31

[0567] Table 14-32

[0568] Table 14-33

[0569] Table 14-34

[0570] Table 14-35

[0571] Table 14-36

[0572] Table 14-37

[0573] Table 14-38

[0574] Table 14-39

[0575] Table 14-40

[0576] Table 14-41

[0577] Table 14-42

[0578] Table 14-43

[0579] Table 14-44

[0580] Table 14-45

[0581] Table 14-46

[0582] Table 14-47

[0583] Table 14-48

[0584]

Table 14-49

[0585] Table 14-50

[0586] Table 14-51

[0587] Table 14-52

[0588] Table 14-53

[0589] [Table 14-54]

[0590] [Table 14-55]

[0591] [Table 14-56]

[0592] [Table 14-57]

[0593] [Table 14-58]

[0594] [Table 14-59]

[0595] [Table 14-60]

[0596] As shown in Table 3 above, the molecular structures of specific exemplary compounds are shown in Figures 1 to 43. It will be served.

[0597] In vitro functional activity Functional activity is measured in GIP-R, GLP-1R, and GcgR-expressing HEK-293 clones. The determination is made in the cell line. Each receptor cell line is subjected to 1×Gluta in a 20 μL assay volume. MAX (trademark) (L-alanyl-L-glutamine dipeptide, Gibco catalog number 3) 5050), 0.1% casein (Sigma catalog number C4765), 1% HSA (Hydrochloride) Serum albumin (Sigma catalog number A3782), 500 μM IBMX (3- Isobutyl-1-methylxanthine, and 20 mM HEPES (4-(2-hydroxy DMEM (Gibco catalog) supplemented with ethyl(1-piperazine ethanesulfonic acid) Example compound (Labcyte Echo Acoustic Liquid) in (G-number 31053) (20-point concentration-response curves prepared with a 2.75-fold serial dilution using a peddler) It will be processed.

[0598] After incubation at 37°C for 30 minutes, the resulting increase in intracellular cAMP was observed in Ci Using sBio's cAMP Dynamic 2 HTRF Assay Kit (62AM4PEJ) It is determined quantitatively using [a specific method]. In short, the cAMP-d2 conjugate in the cell lysis buffer... Next, similarly, the antibody in the cell lysis buffer, anti-cAMP-cAMP-Eu3+-crypto By adding tate, intracellular cAMP levels are detected. The obtained competitive up Incubate the sieve at room temperature for at least 60 minutes, then excite it at 320 nm. The Pherastar instrument (BMG Lab) has emission at 665nm and 620nm. Detected using (tech). Raw data value (665nm / 620nm × 10,000) The emission of cAMP is inversely proportional to the amount of cAMP present, and the raw data values ​​are obtained using the cAMP standard curve. This was then converted to cAMP(nM) per well.

[0599] The amount of cAMP (nM) generated in each well is equivalent to human GLP-1 (7-36) NH2( SEQ ID NO: 1221), Human Glucagon (hGcg) (SEQ ID NO: 1222), or Human GI P (1-42) Percentage of maximum reactions observed with any of NH2 (SEQ ID NO: 1223) It is converted to the maximum reaction percentage fitted to the 4-parameter logistic equation. Using the concentrations of the added peptides against each other, nonlinear regression analysis was performed to determine the relative EC 50 Derive the value ru.

[0600] Exemplary dual agonist conjugates: hGcg, hGIP (1-42) NH2, and hGLP-1 (7-36) Relative EC for NH2 50 The geometric mean of the data is as follows: Table 4 shows examples of triple agonist conjugates, hGcg, and hGIP. (1-42) NH2 and hGLP-1 (7-36) Relative EC for NH2 50 Geometric plane of data The average is shown in Table 5 below.

[0601] [Table 15-1]

[0602] [Table 15-2]

[0603] [Table 15-3]

[0604] [Table 15-4]

[0605] Table 15-5

[0606] Table 15-6

[0607] Table 15-7

[0608] Table 15-8

[0609] Table 15-9

[0610] Table 15-10

[0611] Table 15-11

[0612] Table 15-12

[0613] Table 16-1

[0614] Table 16-2

[0615] [Table 16-3]

[0616] [Table 16-4]

[0617] [Table 16-5]

[0618] [Table 16-6]

[0619] [Table 16-7]

[0620] [Table 16-8]

[0621] Example Compounds in Rats: Plasma Levels The plasma levels of the example compounds were determined in male Sprague Dawley rats. We purchased 8-9 week old animals from Envigo (Indiana, USA). Next, the rats were randomized. Each group (n=5) received 10 nmol via subcutaneous (sc) injection. One compound was administered per kg. Blood samples were taken via the tail vein at the time indicated in each table. The samples were collected continuously. Animal research was conducted at Lilly Institutional Animal Research Center. Approved by the Care and Use Committee (IACUC) .

[0622] Plasma was prepared from collected blood samples. Each plasma sample was prepared using a compound dilution medium. The solution is then serially diluted and subsequently used to express human GLP1R and cAMP-driven luciferase. The cells were incubated with HEK293 cells in a 96-well plate for 5 hours. After dissolving and adding Bio-Glo(Promega), the luminescence signal intensity was measured.

[0623] For each test compound, a mathematical equation is used to calculate the corresponding concentration of the luminescence signal value. Nonlinear regression analysis of quasi-dose-luminescence signal curve data (Graphpad Prism) The dosage in each set of serially diluted samples was established using mathematical equations. The luminescence data showing the dependency was converted to concentration, and the values ​​were averaged as the plasma concentration of the sample. Table 6 The data presented is the mean ± SE of the average values ​​of 5 rats. As can be seen in Table 6. The example compounds tested were, in some cases, subjected to an extended period including at least 28 days. It remained in the plasma for an extended period.

[0624] [Table 17-1]

[0625] [Table 17-2]

[0626] Pharmacokinetic (PK) studies We will conduct PK studies of specific exemplary compounds in cynomolgus monkeys and DIO mice. The compound plasma concentrations for these studies were determined by the ELISA method or as described in more detail below. Determined using the LC / MS method.

[0627] Bioanalytical Methods - ELISA The plasma concentrations of specific compounds were determined by the ELISA method. eptavidin coated plate (Thermo Scientific catalog) (G number 15500) Biotinylated ELI299 2G5 antibody (Lilly-generated antibody lock It was coated using (T number 2G5-22AUG2023). This coating is anti- The body is treated with a pH 9.6 bicarbonate buffer (Thermo Scientific catalog number 2 Dilute to a final concentration of 1 μg / mL (8382) and add to the plate at a rate of 100 μL / well. Then, it was stored overnight at 2-8°C. After overnight incubation, the plate was then placed in a TB (Turban Tablet). Wash using S-Tween washing buffer (TEKNOVA catalog number T0310). All washing steps used the same washing buffer, with a shaking step between each wash, for 30 minutes. This is performed by washing the plate four times with 0 μL / well of washing buffer. After each washing step, gently tap the plate on a paper towel to remove excess washing buffer. After the washing process, the coated and washed plates are used for this assay. Locking and Assay Buffer (Cold in PBS buffer with additional Tween-20) Block using casein buffer - Thermo Scientific Catalog number 37528 was supplemented by adding Tween-20, and 0.05 The final concentration was set to %. This buffer was used as the blocking buffer for the assay and the assay itself. It was used as a buffer. Blocking was performed using 200 μL / well of blocking buffer. The procedure was performed, and the samples were stored at ambient temperature for approximately 1 hour while shaking at 500 RPM. During blocking... Standards, quality control samples, and research samples are mixed in assay buffer in a 1:20 ratio (monkey). Or 1:50 (mouse) MRD (Minimum Required Dilution) The samples were treated through n (minimum required dilution) and any additional sample dilutions as needed. After packing, wash the plate, then fill it with the processed sample in 100 μL / well. It was added to the plate. Then, the plate containing the sample was shaken at 500 RPM, Store at ambient temperature for approximately 1 hour. The detection reagent is mouse anti-human IgG4 Fc-HRP antibody ( This is Southern Biotech catalog number 9040-05. The detection reagent is A Dilute to a final concentration of 0.017 μg / mL in Szey buffer. Incubate the sample. After cleaning, wash the plate and incubate it with 100 μL / well of detection solution. Next, the plate containing the detection reagent is shaken at 500 RPM and left to cool to ambient temperature for approximately 1 minute. Store for a period of time. During detection incubation, the detection substrate solution is treated with KPL TMB peroxide. KPL peroxidase substrate (KPL catalog number 5120-0048) and KPL peroxidase substrate Prepared by mixing an equal volume of solution B (KPL catalog number 5120-0037). After incubation of the detection reagent, wash the plate and add 100 μL / of the detection substrate. Add the solution in the well volume and incubate for 7-8 minutes until color develops. Then stop the TMB. The reaction is stopped using the solution (KPL catalog number 5150-0019). Then, Using a plate reader, read the plate at a wavelength of 450 nm, with a 630 nm correction. .

[0628] Bioanalytical methods - LC / MS The plasma concentrations of the compounds were determined by LC / MS. The incretins and F of the analytes were also analyzed. A substitute peptide for the c subunit was used as a measure of plasma concentration. For each assay, Diluted research samples, blanks, and standards in 100% cynomolgus monkey plasma. Place the sample (95 μL) into a PCR plate (Thermo Scientific 0.2 m³). The sample was transferred to a skirtless 96-well PCR plate (Catalog No. AB-0600). The analyte was pre-conjugated onto streptavidin-coated magnetic beads (100 μL). Biotin-labeled anti-human IgG antibody (Southern Biotech's Goa t Anti-Human IgG, Monkey ads-BIOT, Catalog No. 2 Immunoprecipitation was performed using 049-08). Samples, blanks, and standards were prepared using 1×TBST. After washing once with 1×TBS, the sample was eluted in 0.2% formic acid (100 μL). Elution (95 μL) is used to form an internal structure similar to incretin and Fc subunit (5 μL each). Transfer to a new plate containing the standard material and store under nitrogen at 60°C on an SPE Dual Dry. They were dried. Once dry, the samples, blanks, and standards were subjected to 500 mM ammonium carbonate. Reconstitute in nium (30 μL), then add 2% iodoethanol and 1% triethylphosphine. In acetonitrile (30 μL) containing [the substance], shake at 600 rpm at 40 degrees Celsius. The samples, blanks, and standards were reduced and alkylated for 1 hour. The samples, blanks, and standards were then stored under nitrogen using SPE. After drying twice on a Dual Dry at 60°C, 10% methanol and Glu-C were added. Reconstitution in 50 mM ammonium bicarbonate (95 μL) containing endoproteinase. The mixture was then allowed to digest overnight (approximately 18 hours) at 37°C while being shaken at 600 rpm. Quench with 88% formic acid (5 μL) and use for analysis. Injected into a Sprite Armor C18 column (2.1 × 80 mm, 5 μm) (2 (0 μL). The analyte and internal standard substitute peptide are placed in 0.1% formic acid and acetonitrile water. Thermo Fisher Orbitrap E in 0.1% formic acid at 400 μL / min The substitute peptide was delivered to a Clipse Tribrid mass spectrometer for quantification. Measured using a positive-mode full-scan FT-MS.

[0629] Pharmacokinetics in cynomolgus monkeys Male cynomolgus monkeys were given a compound in 40 mM Tris pH8 buffer at a rate of 0.5 mL / kg. A single subcutaneous dose (10 nmol / kg) was administered. For pharmacokinetic characterization, the drug was administered. From 6, 24, 48, 72, 96, 120, 168, 240, 288, 336, 408, Blood samples were taken from each animal at 576, 672, 2352, 3024, and 4200 hours. Ta.

[0630] Male cynomolgus monkeys were given a compound in 40 mM Tris pH8 buffer at a rate of 0.5 mL / kg. The substance was administered as a single intravenous or subcutaneous dose (10 nmol / kg). Pharmacokinetic characterization was evaluated. Therefore, from administration: 6, 24, 48, 72, 96, 168, 240, 288, 336, 40 8, 480, 576, 672, 840, 1008, 1176, 1344, and 1512 After a short time, blood samples were taken from each animal.

[0631] Female cynomolgus monkeys were given a compound in 40 mM Tris pH8 buffer at a rate of 0.5 mL / kg. The substance was administered as a single intravenous or subcutaneous dose (10 nmol / kg). Pharmacokinetic characterization was evaluated. Therefore, the doses administered were 0.25 (intravenous only), 6, 24, 48, 72, 96, 168, and 24. 0, 288, 336, 408, 480, 576, 672, 840, 1008, 1176, Blood was collected from each animal at 1344, 1512, 1680, 1848, and 2016 hours later. I took it.

[0632] [Table 18] Abbreviation: AUC 0-inf = Area under the curve from 0 time to infinity. CL / F = Clearance S / Bioavailability, T max = Time to reach maximum concentration, C max = Most observed Large plasma concentration, T 1 / 2 =Half-life, NC=Not calculated

[0633] [Table 19] Abbreviation: AUC 0-inf = Area under the curve from 0 time to infinity, CL = clearance, C max = Observed maximum plasma concentration, T 1 / 2 = Half-life

[0634] As can be seen in Tables 7-8, the PK profiles of the example compounds are currently available. This supports the idea that it has a longer elimination half-life and lower clearance compared to cretin therapy. These data suggest that the compounds mentioned above have pharmacokinetic profiles that are potentially suitable for monthly administration. It demonstrates that they possess it.

[0635] Pharmacodynamics in DIO mice The in vivo efficacy of specific example compounds on body weight was observed in a diet-induced obesity mouse model (diet -Induced obesity (DIO) was determined. Male C57BL / 6DIO mice (The Jackson La) maintained in (contains) The boratory (Bar Harbor, ME) was used in the following study. The mice were: Individually placed in a temperature-controlled facility (24°C) with a 12-hour light / dark cycle (lights on at 22:00). They were confined and had free access to food and water. To investigate the effect on body weight, obese Mau In the vehicle (40 mM Tris-HCl at pH 8.0) or test substance (3, 10 and A single subcutaneous dose of 30 nmol / kg was administered. Body weight and food intake were monitored for the first 14 days. The desired long duration of action and one When each example compound is administered as a single dose to treat obese animals, sustained weight loss is observed. I was made to do it.

[0636] [Table 20-1]

[0637] [Table 20-2]

[0638] [Table 20-3]

[0639] [Table 20-4]

[0640] Body weight dose-response study in obese mice The in vivo effects of the example compounds on body weight and food intake were observed in a diet-induced obese mouse model. It was decided by DIO. A high-calorie diet (containing 40% kcal from fat) is used for maintenance. The male C57BL / 6 obese mice were subjected to a 12-hour light / dark cycle (lights turned on at 22:00). They were housed in a temperature-controlled facility (24°C) and had free access to food and water. Compound 1 To investigate the dose-dependent effects on body weight in 70, 172, 387, and 456 individuals with obesity, The animals were given either a vehicle (40 mM Tris-HCl at pH 8.0) or a test substance (0. The test substance was administered subcutaneously to obese mice at concentrations of 3, 1, 3, 10, and 10 nmol / kg. Treatment resulted in a dose-dependent reduction in body weight compared to control animals (Figures 44-47). The results are presented as mean ± SEM, with n=6 per group. Statistical analysis is performed using two-way ANOVA. Next, Dunnett's method for multiple comparisons was used. A statistically significant difference was confirmed with p<0.05. It was confirmed. Overall, these findings indicate that the example compounds result in a definite reduction in weight. This indicates that.

Claims

1. formula: 【Chemistry 1】 A compound of or a pharmaceutically acceptable salt thereof, wherein the formula is X ＴＰＰ It is a therapeutic polypeptide conjugated to a fatty acid portion, Z is O, C(O), NH, C １ ~C ３０ Alkyl, (OCH ２ CH ２ ) ｍ [C(O )NH-CH ２ CH ２ OCH ２ ｍ , (2-[2-(2-amino-ethoxy)-ethoxy​ ]-acetyl) ｍ It comprises amino acids, peptides, or combinations thereof, and m is 1 to 30 It is an integer, U is C １ ~C ５ Alkyl, N, phenyl, or phenylcarbonyl, or these Includes combinations, R １ and R ２ It is either independent, nonexistent, covalently bonded, or C １ ~C ５ Alki It is R ３ and R ４ It is either independent, nonexistent, or an amide. R ５ and R ６ It is either independent, does not exist, or R ５ and R ６ Independently, C １ ~C ５ Alkyl, phenyl, CH ２ O(CH ２ CH ２ O) ｎ , or a combination of these Including n, n is an integer from 1 to 5, C １ ~C ５ Alkyl is NH ２、 CH ２ NH ２ , and C H ２ CH ２ NH ２ Replaced by any one of the following: R ７ and R ８ It is either independent, does not exist, or 【Chemistry 2】 Selected from the group consisting of, in the formula, ( ＊ ) is R ９ or R １０ Includes a connection point, ( ＊＊ ) is R ５ or R ６ Includes connection points, R ９ and R １０ This is independently an antibody fragment containing an Fc region, a compound or its pharmaceutical Salts that are permissible.

2. R ７ and R ８ but, 【Transformation 3】 The compound according to claim 1 or a pharmaceutically acceptable salt thereof.

3. R ５ and R ６ However, each of them is NH ２ C replaced by ２ It is alkyl, as described in claim 2. A compound of or a pharmaceutically acceptable salt thereof.

4. R ３ and R ４ However, each of them is an amide, according to any one of claims 1 to 3 A substance or its pharmaceutically acceptable salt.

5. R １ and R ２ However, each of them is C ２ Alkyl, as described in any one of claims 1 to 4 A compound of or a pharmaceutically acceptable salt thereof.

6. A compound or drug according to any one of claims 1 to 5, wherein U is N or phenyl. A scientifically acceptable salt.

7. The aforementioned compound, 【Chemistry 4】 A compound according to claim 6 or a pharmaceutically acceptable salt thereof, comprising the above.

8. R ７ and R ８ but, 【Transformation 5】 The compound according to claim 1 or a pharmaceutically acceptable salt thereof.

9. R ５ and R ６ However, phenyl and CH are respectively ２ O(CH ２ CH ２ O) ｎ Including n is The compound according to claim 8 or a pharmaceutically acceptable salt thereof, which is 2.

10. R ３ and R ４ However, each is an amide, as described in any one of claims 8 to 9 A substance or its pharmaceutically acceptable salt.

11. R １ and R ２ However, each of them is C ２ Alkyl, as described in any one of claims 8 to 10 The listed compounds or their pharmaceutically acceptable salts.

12. A compound according to any one of claims 8 to 11, wherein U is N or phenyl, or the Pharmaceutically acceptable salts.

13. The aforementioned compound, 【Transformation 6】 A compound according to claim 12 or a pharmaceutically acceptable salt thereof, comprising the above.

14. R ７ and R ８ However, each of them, 【Transformation 7】 The compound according to claim 1 or a pharmaceutically acceptable salt thereof.

15. R ５ and R ６ However, phenyl and CH are respectively ２ O(CH ２ CH ２ O) ｎ Including n is The compound according to claim 14 or a pharmaceutically acceptable salt thereof, which is 2.

16. R ３ and R ４ However, each is an amide, as described in any one of claims 14 to 15. A compound or a pharmaceutically acceptable salt thereof.

17. R １ and R ２ However, each of them is C ２ Alkyl, according to any one of claims 14 to 16 The compound or a pharmaceutically acceptable salt thereof.

18. The compound according to any one of claims 14 to 17, wherein U is N or phenyl. A pharmaceutically acceptable salt of [the substance].

19. The aforementioned compound, 【Transformation 8】 A compound according to claim 18 or a pharmaceutically acceptable salt thereof, comprising the above.

20. R ７ and R ８ However, each of the compounds described in claim 1 or their pharmaceutically acceptable compounds does not exist. Salt is acceptable.

21. R ５ and R ６ However, each of them is C １ The compound according to claim 1 or 20, which is alkyl or This is the pharmaceutically acceptable salt.

22. R ３ and R ４ are each amido, the compound according to claim 21 or a pharmaceutically Salts that are permissible.

23. R １ and R ２ However, each of them is C ２ Alkyl, according to any one of claims 20 to 22 The compound or a pharmaceutically acceptable salt thereof.

24. The compound according to any one of claims 20 to 23, wherein U is N or phenyl. A pharmaceutically acceptable salt of [the substance].

25. The aforementioned compound, 【Chemistry 9】 A compound according to claim 24 or a pharmaceutically acceptable salt thereof, comprising the above.

26. The aforementioned compound, 【Chemistry 10】 A compound according to claim 1 or a pharmaceutically acceptable salt thereof, comprising the above.

27. Z is (OCH ２ CH ２ ) ２４ -NH-C(O)CH ２ CH ２ , according to claims 1 to 26 A compound or a pharmaceutically acceptable salt thereof as described in any one of the items.

28. Z has 1 to 8 (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl) moieties. A compound according to any one of claims 1 to 27, or a pharmaceutically acceptable salt thereof, comprising [a specific component]. 。

29. Claim 1, wherein Z comprises a peptide containing any of the sequence numbers 1182 to 1217. A compound described in any one of items 28 or a pharmaceutically acceptable salt thereof.

30. The therapeutic polypeptide is one of GIP, GLP-1, and glucagon receptor The compound or the compound according to any one of claims 1 to 29, which includes an agonist as described above. Pharmaceutically acceptable salts.

31. The aforementioned therapeutic polypeptide X １ X ２ X ３ X ４ TX ６ TSDX １０ X １１ X １２ X １３ LX １５ X １６ KAX １９ X ２０ X ２１ FIX ２４ X ２５ LX 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 X ４０ X ４１ (Sequence ID 1218) X １ is H, NMey, or Y, X ２ These are Ac4c, Aib, αMeS, Iva, or D-Ala. X ３ is Q, E, or H, X ４ It is G or D-Ala. X ６ is αMeF(2F), F, or αMeF, X １０ is 4-Pale, Y, or V, X １１ is Aib, S, or αMeS, X １２ is either I or S, X １３ is L or αMeL, X １５ is either D or E, X １６ is K, E, or Orn, X １９ is A or Q, X ２０ These are Aib, αMeL, Iva, or αMe4Paal. X ２１ is E, Q, D, or Orn, X ２４ These are K, Q, E, D-Glu, or D-Glun. X ２５ is αMeY, W, or Y, X ２７ is I, L, or V, X ２８ is K, E, or A, X ２９ is Aib, G, S, or Q, X ３０ is G, H, or S, X ３１ is P, G, E, or Orn, X ３２ It does not exist, or it is K, S, or P. X ３２ If S or P, then X ３３ is S, X ３３ If S, then X ３４ is G or Aib, X ３４ If is G or Aib, then X ３５ It does not exist, or it is A or Orn. the law of nature, X ３５ If is A or Orn, then X ３６ It does not exist, or it is P. X ３６ If P, then X ３７ It does not exist, or it is P. X ３７ If P, then X ３８ It does not exist, or it is P or Ac3c. X ３８ If is P or Ac3c, then X ３９ It does not exist, or S, Orn, or It is G, X ３９ If is S, Orn, or G, then X ４０ It does not exist, or it is K or G. X ４０ If is K or G, then X ４１ It does not exist, or it is S or G. X ３２ If X does not exist or is K, ３３ ~X ４１ It does not exist X ３５ If X does not exist, ３６ ~X ４１ It does not exist, X ３６ If X does not exist, ３７ ~X ４１ It does not exist, X ３７ If X does not exist, ３８ ~X ４１ It does not exist, X ３８ If X does not exist, ３９ ~X ４１ It does not exist, X ３９ If X does not exist, ４０ and X ４１ It does not exist, X ４０ If X does not exist, ４１ It does not exist, as described in any one of claims 1 to 30. A compound of or a pharmaceutically acceptable salt thereof.

32. X ２４ , X ２８ , X ３２ or X ４０ One of them is conjugated to Z, please The compound described in item 31 or a pharmaceutically acceptable salt thereof.

33. The therapeutic polypeptide is one of the following: SEQ ID NOs: 1-186 or 996-1038 A compound according to any one of claims 1 to 31, comprising 90% sequence identity with any of the above. or a pharmaceutically acceptable salt thereof.

34. The therapeutic polypeptide is one of the following: SEQ ID NOs: 1-186 or 996-1038 A compound according to any one of claims 1 to 32, or a pharmaceutically acceptable compound thereof, comprising any one of the above. Salt.

35. The aforementioned fatty acid portion is C １６ ~C ２２ A fatty acid as described in any one of claims 1 to 32. The listed compounds or their pharmaceutically acceptable salts.

36. The aforementioned fatty acid is conjugated to the therapeutic polypeptide at the K17 position via a linker. The compound according to claim 35 or a pharmaceutically acceptable salt thereof.

37. The linker comprises 1 to 4 amino acids, the compound according to claim 36 or the pharmaceutical thereof. A generally acceptable salt.

38. The linker contains one or more of the amino acids Lys, εLys, Glu, and γGlu. , or a combination thereof, the compound according to claim 37 or a pharmaceutically acceptable thereof Salt.

39. The linker has 1 to 8 (2-[2-(2-amino-ethoxy)-ethoxy]- A compound according to any one of claims 35 to 38, including the cetyl portion, or pharmaceutically thereof Allowable salt.

40. The linker and the fatty acid are (εLys) ａ (γGlu) ｂ - (2 - [2 - (2 - (amino-ethoxy)-ethoxy)-acetyl) ｃ -CO-(CH ２ ) ｐ -CO ２ Structure of H The formula includes, where a is an integer of 0, 1, or 2, b is an integer of 0 or 1, and c is an integer of 1 to 8. The integer, where p is an integer between 14 and 18, according to any one of claims 35 to 39. A compound or a pharmaceutically acceptable salt thereof.

41. The therapeutic polypeptide conjugated to the fatty acid portion corresponds to Sequence IDs 187-458. or a claim comprising 90% sequence identity with any of sequence numbers 1039 to 1098. A compound described in any one of items 35 to 40, or a pharmaceutically acceptable salt thereof.

42. The therapeutic polypeptide conjugated to the fatty acid portion corresponds to Sequence IDs 187-458. Or any one of claims 1 to 40, including any one of sequence numbers 1039 to 1098. The compounds listed in the section or their pharmaceutically acceptable salts.

43. The Fc region is the sequence numbers 935-936, 941-948, 1178-1181 A compound or drug according to any one of claims 1 to 42, selected from any one of the above. A scientifically acceptable salt.

44. The compound is derived from any of the sequence numbers 459-934 or 1099-1177. A selected compound according to claim 1 or a pharmaceutically acceptable salt thereof.

45. formula: 【Chemistry 11】 A compound of or a pharmaceutically acceptable salt thereof, wherein the formula is X ＴＰＰ It is a therapeutic polypeptide, YAibEGTX ６ TSDX １０ X １１ X １２ X １３ LX １５ X １６ KAQX ２０ X ２ １ FIX ２４ X ２５ LX 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 X ４０ X ４１ (Sequence ID 1219) X ６ is αMeF(2F), F, or αMeF, X １０ is 4-Pale, Y, or V, X １１ is Aib or S, X １２ is either I or S, X １３ is L or αMeL, X １５ is either D or E, X １６ is E or Orn, X ２０ These are Aib, αMeL, or Iva, X ２１ is E, Q, D, or Orn, X ２４ is K, Q, E, or D-Glu, X ２５ is αMeY or Y, X ２７ is I, L, or V, X ２８ is K, E, or A, X ２９ is Aib, G, or Q, X ３０ is G or S, X ３１ is P, G, E, or Orn, X ３２ It does not exist, or it is K, S, or P. X ３２ If S or P, then X ３３ is S, X ３３ If S, then X ３４ is G or Aib, X ３４ If is G or Aib, then X ３５ It does not exist, or it is A or Orn. X ３５ If is A or Orn, then X ３６ It does not exist, or it is P. X ３６ If P, then X ３７ It does not exist, or it is P. X ３７ If P, then X ３８ It does not exist, or it is P. X ３８ If P, then X ３９ It does not exist, or it is S, Orn, or G. X ３９ If is S, Orn, or G, then X ４０ It does not exist, or it is K or G. X ４０ If is K or G, then X ４１ It does not exist, or it is S or G. X ３２ If X does not exist or is K, ３３ ~X ４１ It does not exist X ３５ If X does not exist, ３６ ~X ４１ It does not exist, X ３６ If X does not exist, ３７ ~X ４１ It does not exist, X ３７ If X does not exist, ３８ ~X ４１ It does not exist, X ３８ If X does not exist, ３９ ~X ４１ It does not exist, X ３９ If X does not exist, ４０ and X ４１ It does not exist, X ４０ If X does not exist, ４１ It does not exist. L is R ９ , R １０ and phosphorus conjugated to each of the therapeutic polypeptides. It is a car, R ９ and R １０ This is an antibody fragment containing an Fc region, independently of the others. X ２４ , X ２８ , X ３２ or X ４０ One of them is conjugated to the linker A compound or a pharmaceutically acceptable salt thereof.

46. The therapeutic polypeptide is a dual agonist at the GIP receptor and the GLP-1 receptor. A compound according to claim 45 or a pharmaceutically acceptable salt thereof that is active.

47. X １ Y is X ２ is Aib, and X ３ is E and X ４ G is X ６ αMeF(2F) is X １０ is 4-Pal or Y, and X １１ is Aib or S, X １２ is I and X １３ αMeL and X １５ It is D, X １６ is E or Orn, and X １ Q is X ２０ is Aib, and X ２１ teeth , E and X ２４ is Q, E, or D-Glu, and X ２５ αMeY and X ２７ is I and X ２８ is E and X ２９ is Aib or G, and X ３０ G or S, X ３１ P is X ３２ is G or S, X ３２ If S, then X ３３ is S, X ３４ G is X ３５ A is X ３６ P is X ３７ P is X ３８ P is X ３９ is S, X ４０ is K X ４１ It does not exist, and the linker is X ４０ Conjugated to, X ３２ If G, then X ３３ ~X ４１ It does not exist, and the linker is X ３２ ni Konju The compounds according to any one of claims 45 to 46, each comprising being gated or The pharmaceutically acceptable salt.

48. X １０ is Y or 4-Pal, and X １１ is Aib or S, and X ２４ Q, E, or D-Glu, and X ３０ is at least one of G or S. , the compound according to claim 47 or a pharmaceutically acceptable salt thereof.

49. X １０ is Y or 4-Pal, and X １１ is Aib, and X ２４ E is, X ３０ This includes S and X ４０ is K, and the linker has a conjugate The compound according to claim 48 or a pharmaceutically acceptable salt thereof.

50. The therapeutic polypeptide is one of SEQ ID NOs: 1 to 92 or 996 to 999. A compound according to any one of claims 45 to 49, which includes 90% sequence identity. This is the pharmaceutically acceptable salt.

51. The therapeutic polypeptide is any of numbers 1 to 92 or sequence numbers 996 to 999. A compound according to any one of claims 45 to 50, including or the pharmaceutically acceptable thereof salt.

52. The aforementioned therapeutic polypeptide is sequence numbers 1, 2, 22, 43, 45, 48, 55, 56, Claims 45 to 51 include any one of 58, 59, 61, 66, 68, and 47. A compound listed in any one of the items, or a pharmaceutically acceptable salt thereof.

53. The therapeutic polypeptide is linked at position 17 via a second linker. １６ ~C ２２ fatty acid part The compound according to any one of claims 45 to 51 or the compound conjugated in minutes Pharmaceutically acceptable salts.

54. The compound according to claim 53, wherein the second linker comprises 1 to 4 amino acids or the same Pharmaceutically acceptable salts.

55. The second linker is the amino acids Lys, ε-Lys, Glu, γ-Glu or these A compound according to claim 54, including combinations thereof, or a pharmaceutically acceptable salt thereof.

56. The aforementioned second linker has 1 to 8 (2-[2-(2-amino-ethoxy)-ethoxy] A compound or pharmaceutically thereof according to any one of claims 53 to 55, comprising the -acetyl) moiety. A generally acceptable salt.

57. The second linker and the fatty acid are (ε-Lys) ａ (γGlu) ｂ - (2 - [2 - (2-amino-ethoxy)-ethoxy-acetyl) ｃ -CO-(CH ２ ) ｐ -CO ２ H The structure includes, where a is an integer 0, 1 or 2, b is an integer 0 or 1, and c is The integer is between 1 and 8, and p is an integer between 14 and 18, according to any one of claims 53 to 56. The compound or a pharmaceutically acceptable salt thereof.

58. The therapeutic polypeptide conjugated to the fatty acid portion via the second linker Chido is paired with one of the sequence numbers 187-332 or 1039-1042. A compound or pharmaceutically thereof according to any one of claims 53 to 57, including 0% sequence identity A generally acceptable salt.

59. The therapeutic polypeptide conjugated to the fatty acid portion via the second linker The cido contains one of the sequence numbers 187-332 or 1039-1042. The compound according to any one of claims 53 to 58 or a pharmaceutically acceptable salt thereof.

60. The therapeutic polypeptide conjugated to the fatty acid portion is sequence number 187, 2 ０９、２３５、２３７、２４１、２４３、２５７、２５８、２６０、２６２、２６３、２ Claims 53 to 59 include any one of 64, 266, 267, 275, or 277. A compound or a pharmaceutically acceptable salt thereof as described in any one of the items.

61. The Fc region is the one corresponding to sequence numbers 935-936, 941-948, or 1178-1181. A compound or so selected from any one of claims 45 to 60. A pharmaceutically acceptable salt of [the substance].

62. The linker, 【Chemistry 12】 Including, in the formula, Z is O, C(O), NH, C １ ~C ３０ Alkyl, (OCH ２ CH ２ ) ｍ [C(O ) NH-CH ２ CH ２ OCH ２ ] ｍ This includes peptides, or combinations thereof, where m is 1 It is an integer up to 30, ( ＊＊＊ ) includes a connection point to the therapeutic polypeptide, U is C １ ~C ５ Alkyl, N, phenyl, or phenylcarbonyl, or these Includes combinations, R １ and R ２ These can be independent, covalently bonded, or C １ ~C ５ It is alkyl, R ３ and R ４ It does not exist, or it is an amide. R ５ and R ６ It is either independent, does not exist, or R ５ and R ６ Independently, C １ ~C ５ Alkyl, phenyl, CH ２ O(CH ２ CH ２ O) ｎ , or a combination of these Including n, n is an integer from 1 to 5, C １ ~C ５ Alkyl is NH ２、 CH ２ NH2, and C H ２ CH ２ NH ２ Replaced by any one of the following: R ７ and R ８ It does not exist, or independently, 【Chemistry 13】 Selected from the group consisting of, in the formula, ( ＊ ) is R ９ or R １０ Includes a connection point, ( ＊＊ ) is R ５ or R ６ Includes connection point , the compound according to any one of claims 45 to 61 or a pharmaceutically acceptable salt thereof.

63. The compound according to claim 62, or the pharmaceutically acceptable thereof, wherein U is N or phenyl. salt.

64. R ７ or R ８ but, 【Chemistry 14】 The compound according to claim 62 or 63, or a pharmaceutically acceptable salt thereof.

65. The linker, 【Chemistry 15】 Includes, In the formula, ( ＊ ) is R ９ or R １０ A compound according to claim 64 or the same, including a connection point Pharmaceutically acceptable salts.

66. R ７ and R ８ but, 【Chemistry 16】 The compound according to claim 62 or a pharmaceutically acceptable salt thereof.

67. The linker, 【Chemistry 17】 Includes, In the formula, ( ＊ ) is R ９ or R １０ The compound according to claim 66 or the connection point thereof Pharmaceutically acceptable salts.

68. Z is (OCH ２ CH ２ ) ２４ -NH-C(O)CH ２ CH ２ Claims 62 to 6, including A compound as described in any one of item 7.

69. Z has 1 to 8 (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl) moieties. A compound according to any one of claims 65 to 68, including a portion thereof, or a pharmaceutically acceptable compound thereof. salt.

70. Claim 6, wherein Z comprises a peptide containing any of the sequence numbers 1182 to 1217. A compound described in any one of items 5 to 69, or a pharmaceutically acceptable salt thereof.

71. The compound is one of the sequence numbers 459-787 or 1099-1100 The compound described in any one of claims 45 to 70 or the pharmaceutically acceptable compound thereof. Salt.

72. The aforementioned compounds are sequence numbers 459, 490, 505, 585, 587, 594, 616, ６２２、６２６、６２７、６２８、６３０、６３１、６３９、６４１、６４２、６４３、 It is one of the following: 645, 646, 662, 698, 699, 700, or 1100. , the compound according to any one of claims 45 to 71 or a pharmaceutically acceptable salt thereof.

73. formula: [Chemistry 18] A compound of or a pharmaceutically acceptable salt thereof, wherein the formula is X ＴＰＰ It is a therapeutic polypeptide, X １ X ２ X ３ X ４ TX ６ TSDX １０ X １１ X １２ X １３ LX １５ X １６ KAQX ２０ X ２１ FIX ２４ X ２５ LX 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 X ４０ X ４１ (Sequence ID 1220) X １ is H, NMey, or Y, X ２ These are Ac4c, Aib, αMeS, Iva, or D-Ala. X ３ is Q, E, or H, X ４ It is G or D-Ala. X ６ is αMeF(2F), F, or αMeF, X １０ is 4-Pale, Y, or V, X １１ is Aib, S, or αMeS, X １２ is either I or S, X １３ is L or αMeL, X １５ is either D or E, X １６ is E or Orn, X ２０ These are Aib, αMeL, Iva, or αMe4Paal. X ２１ is E, Q, D, or Orn, X ２４ These are K, Q, E, D-Glu, or D-Glun. X ２５ is αMeY or Y, X ２７ is I, L, or V, X ２８ is K, E, or A, X ２９ is Aib, S, G, or Q, X ３０ is G, S, or H, X ３１ is P, G, E, or Orn, X ３２ It does not exist, or it is K, S, or P. X ３２ If S or P, then X ３３ is S, X ３３ If S, then X ３４ is G or Aib, X ３４ If is G or Aib, then X ３５ It does not exist, or it is A or Orn. the law of nature, X ３５ If is A or Orn, then X ３６ It does not exist, or it is P. X ３６ If P, then X ３７ It does not exist, or it is P. X ３７ If P, then X ３８ It does not exist, or it is P. X ３８ If P, then X ３９ It does not exist, or it is S, Orn, or G. X ３９ If is S, Orn, or G, then X ４０ It does not exist, or it is K or G. X ４０ If is K or G, then X ４１ It does not exist, or it is S or G. X ３２ Or, if K does not exist, X ３３ ~X ４１ It does not exist, X ３５ If X does not exist, ３６ ~X ４１ It does not exist, X ３６ If X does not exist, ３７ ~X ４１ It does not exist, X ３７ If X does not exist, ３８ ~X ４１ It does not exist, X ３８ If X does not exist, ３９ ~X ４１ It does not exist, X ３９ If X does not exist, ４０ and X ４１ It does not exist, X ４０ If X does not exist, ４１ It does not exist. L is R ９ , R １０ , and the ri conjugated to each of the therapeutic polypeptides It is a nker, R ９ and R １０ These are antibody fragments containing an Fc region, X ２４ , X ３２ and X ４０ One of them is conjugated to the linker, A compound or a pharmaceutically acceptable salt thereof.

74. The therapeutic polypeptide triples in GIP, GLP-1, and glucagon receptors. A compound according to claim 73 or a pharmaceutically acceptable salt thereof having agonist activity.

75. X １ is H or Y, and X ２ is Aib, and X ３ Q is X ４ It is G ri, X ６ αMeF(2F) is X １０ is Y or 4-Pal, and X １１ teeth, S or αMeS, X １２ is I and X １３ αMeL and X １５ D X ２０ is either Aib or αMe4Paal, and X ２１ is E or Orn, X ２４ is E or d-Glu, and X ２５ αMeY and X ２７ is L or I Yes, X ２８ is E and X ２９ ~X ３９ is GSPSGAPPPS, X ４０ The compound according to claim 73 or 74, or the pharmaceutically equivalent thereof, comprising K Allowable salt.

76. X １０ is Y or 4-Pal, and X ２４ is E or d-Glu, and X ３０ teeth, The compound according to claim 75, or the pharmaceutically equivalent thereof, comprising at least one of S Allowable salt.

77. X １０ is Y or 4-Pal, and X ２４ is E or d-Glu, and X ３０ teeth, S is X ４０ Each of the following is K, which is conjugated to the linker. Including the compound described in claim 76 or a pharmaceutically acceptable salt thereof.

78. The therapeutic polypeptide is the one specified in SEQ ID NOs. 93-186 or SEQ ID NOs. 1000-1038. The following is a description of any one of claims 73 to 77, which includes 90% sequence identity with any one of the above. A compound or a pharmaceutically acceptable salt thereof.

79. The therapeutic polypeptide is the one specified in SEQ ID NOs. 93-186 or SEQ ID NOs. 1000-1038. A compound or its pharmaceutically acceptable chemical, including any of the compounds described in any one of claims 73 to 78. Salts that are permissible.

80. The aforementioned therapeutic polypeptides are sequence numbers 97, 113, 128, 130, 144, and 171. , a compound according to any one of claims 73 to 79, comprising any of 183 or The pharmaceutically acceptable salt.

81. The therapeutic polypeptide, via a second linker, １６ ~C ２２ fatty acid A compound according to any one of claims 73 to 80, which is partially conjugated or The pharmaceutically acceptable salt.

82. The compound according to claim 81 or the second linker comprising 1 to 4 amino acids Pharmaceutically acceptable salts.

83. The second linker is one of the amino acids Lys, εLys, Glu, and γGlu. The compounds described in claim 82, or any combination thereof, including the above, or any combination thereof, or pharmaceutically acceptable compounds thereof. Salt.

84. The aforementioned second linker has 1 to 8 (2-[2-(2-amino-ethoxy)-ethoxy] A compound or pharmaceutically thereof according to any one of claims 81 to 83, comprising the acetyl moiety. A generally acceptable salt.

85. The second linker and the fatty acid are (ε-Lys) ａ (γGlu) ｂ - (2 - [2 - (2-amino-ethoxy)-ethoxy-acetyl) ｃ -CO-(CH ２ ) ｐ -CO ２ H The structure includes, where a is an integer 0, 1 or 2, b is an integer 0 or 1, and c is The integer is between 1 and 8, and p is an integer between 14 and 18, according to any one of claims 81 to 84. The compound or a pharmaceutically acceptable salt thereof.

86. The therapeutic polypeptide conjugated to the fatty acid portion via the second linker Chido is paired with one of the sequence numbers 333-458 or 1043-1098. A compound or pharmaceutically thereof according to any one of claims 81 to 85, including 0% sequence identity A generally acceptable salt.

87. The therapeutic polypeptide conjugated to the fatty acid portion via the second linker The cydo contains one of the sequence numbers 333-458 or 1043-1098. The compound according to any one of claims 81 to 86 or a pharmaceutically acceptable salt thereof.

88. The therapeutic polypeptide conjugated to the fatty acid portion via the second linker Chido, sequence numbers 337, 380, 395, 404, 423, 424, 426, 435, Claims 81 to 87 include any one of 447, 450, 467, 490, or 493. A compound or a pharmaceutically acceptable salt thereof as described in any one of the items.

89. The Fc region is the one corresponding to sequence numbers 935-936, 941-948, or 1178-1181. A compound or the compound according to any one of claims 73 to 88, selected from any one of the above. A pharmaceutically acceptable salt of [the substance].

90. The linker, 【Chemistry 19】 Including, in the formula, Z is O, C(O), NH, C １ ~C ３０ Alkyl, (OCH ２ CH ２ ) ｍ [C(O ) NH-CH ２ CH ２ OCH ２ ] ｍ This includes peptides, or combinations thereof, where m is 1 It is an integer up to 30, ( ＊＊＊ ) includes a connection point to the therapeutic polypeptide, U is C １ ~C ５ Alkyl, N, phenyl, or phenylcarbonyl, or these Includes combinations, R １ and R ２ These can be independent, covalently bonded, or C １ ~C ５ It is alkyl, R ３ and R ４ It does not exist, or it is an amide. R ５ and R ６ It is either independent, does not exist, or R ５ and R ６ Independently, C １ ~C ５ Alkyl, phenyl, CH ２ O(CH ２ CH ２ O) ｎ , or a combination of these Including n, n is an integer from 1 to 5, C １ ~C ５ Alkyl is NH ２ ,CH ２ NH2, and C H ２ CH ２ NH ２ Replaced by any one of the following: R ７ and R ８ It does not exist, or independently, 【Chemistry 20】 Selected from the group consisting of, in the formula, ( ＊ ) is R ９ or R １０ Includes a connection point, ( ＊＊ ) is R ５ or R ６ Includes connection point , the compound according to any one of claims 73 to 89 or a pharmaceutically acceptable salt thereof.

91. The compound according to claim 90 or the pharmaceutically acceptable thereof, wherein U is N or phenyl. salt.

92. R ７ or R ８ but, 【Chemistry 21】 The compound according to claim 90 or 91 or a pharmaceutically acceptable salt thereof.

93. The linker, 【Chemistry 22】 Includes, in the formula, ( ＊ ) is R ９ and R １０ Any of claims 90 to 92, including a connection point to The compounds described in item 1 or their pharmaceutically acceptable salts.

94. R ７ and R ８ but, 【Chemistry 23】 The compound according to claim 90 or a pharmaceutically acceptable salt thereof.

95. The linker, 【Chemistry 24】 Includes, In the formula, ( ＊ ) is R ９ or R １０ A compound according to claim 90 or the same, including a connection point Pharmaceutically acceptable salts.

96. Z is (OCH ２ CH ２ ) ２４ -NH-C(O)CH ２ CH ２ Claims 90 to 9, including A compound or a pharmaceutically acceptable salt thereof as described in any one of item 5.

97. Z has 1 to 8 (2-[2-(2-amino-ethoxy)-ethoxy]-acetyl) moieties. A compound according to any one of claims 90 to 96, including a portion thereof, or a pharmaceutically acceptable compound thereof. salt.

98. Claim 9, wherein Z comprises a peptide containing any of the sequence numbers 1182 to 1217. A compound listed in any one of items 0 to 96.

99. The compound is one of the sequence numbers 788-934 or 1101-1177 The compound according to any one of claims 73 to 98 or the pharmaceutically acceptable compound thereof. Salt.

100. The aforementioned compounds are sequence numbers 792, 843, 859, 869, 889, 891, 900, ９１２、９１５、１１０１、１１０２、１１０３、１１０４、１１１０、１１７２、１１ Any one of claims 73 to 99, which is 73, 1174, or 1175. The compounds described or their pharmaceutically acceptable salts.

101. The compound has a sufficiently extended duration of action to allow for administration once a month. , the compound according to any one of claims 1 to 100 or a pharmaceutically acceptable salt thereof.

102. A compound according to any one of claims 1 to 101 or a pharmaceutically acceptable salt thereof, and A pharmaceutical composition comprising at least one pharmaceutically acceptable carrier, diluent, or excipient.

103. The pharmaceutical composition according to claim 102, wherein the composition is formulated for subcutaneous administration.

104. A method for treating a disease or condition, wherein an effective amount is described in any one of claims 1 to 101. The compound listed or a pharmaceutically acceptable salt thereof is used to treat the disease or condition described above. A method comprising the step of administering a substance to an individual once a month.

105. The aforementioned disease or condition is diabetes, obesity, chronic weight management, non-alcoholic fatty liver disease ( NAFLD, non-alcoholic steatohepatitis (NASH), dyslipidemia, metabolic syndrome Drome, chronic kidney disease (CKD), osteoarthritis (OA), obesity-related sleep apnea (OS) A) Polycystic ovary syndrome (PCOS), Parkinson's disease, Alzheimer's disease, heart failure ( High heart rate (HF), hypertension (HT), peripheral artery disease (PAD), metabolic syndrome, chronic lumbar spine Comprising of pain (CLBP), cirrhosis, polycystic ovary syndrome, and / or alcohol abuse disorder. The method according to claim 104, selected from the group.

106. The aforementioned pharmaceutically acceptable salts are sodium, potassium, trifluoroacetate, hydrochloride, A compound or drug according to any one of claims 1 to 101, selected from or an acetate salt. A scientifically acceptable salt.

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