Macrocycle-containing glucose sensitive peptides

Abstract

A molecular conjugate comprising a glucose binding macrocycle as represented below that is covalently bonded directly or via linkers to a polypeptide such as insulin to form a glucose sensing insulin (GSI) are described herein. The disclosure includes pharmaceutical compositions, use and method of treatments with such GSIs. Substituents and the various moieties of the structures below are further specified herein. (II) or (III)

Description

[0001] MACROCYCLE-CONTAINING GLUCOSE SENSITIVE PEPTIDES

[0002] BACKGROUND

[0003] Glucose-sensitive insulin (“GSF’) is less active at low blood glucose and more active at high glucose. Various concepts for glucose-sensitive insulin have been pursued. One idea involves insulin conjugated with both a glucose binder plus a binding partner. At low glucose concentration, the two motifs will bind across insulin resulting in reduced insulin bioactivity, while at higher glucose concentration, the binding partner is competed out by glucose, leading to higher insulin bioactivity at higher glucose levels. (For a comprehensive review see Thomas Hoeg-Jensen Glucose-sensitive insulin, April 2021, Molecular Metabolism 46 pp. 101-107 available at https: / / doi.org / 10.1016 / j.molmet.2020.101107). This idea has been pursued mainly by using boronates as glucose binders.

[0004] Another idea for a glucose binder is a specific glucose binding non-boronate macrocycle. For example, a glucose binder macrocycle was previously conjugated to B29 of insulin and a glucoside to Bl, via specific linkers. (Thomas Hoeg-Jensen, Rita Slaaby, Thomas Kruse et al. Glucose-sensitive insulin with attenuation of hypoglycaemia, 09 May 2023, PREPRINT (Version 1) available at Research Square [https: / / doi.org / 10.21203 / rs.3.rs-2882397 / vl]). Notably, the given macrocycle is relatively elaborate to synthesize, and it binds moderately strongly to albumin, which may compete with glucose binding and affect insulin bioactivity. Also, for a meal-type glucose sensitive insulin (i.e. prandial insulin), fast insulin absorption from the injection site is desired, but binding to albumin at the injection site can slow the absorption. Therefore, there is a desire to develop other approaches for GSIs with smaller macrocycles and possibly without boronates to provide sufficient glucose sensitivity and desirable speed of action.

[0005] Another macrocyle with glucose binding activity was recently presented (Dr. Canjia Zhai, Chengkai Xu, Wenqi Liu et. al. Dynamic Approach to Synthetic Lectin for Glucose with Boosted Binding Affinity through C-H Hydrogen Bonds, March 13 2023, Chemistry A European Journal available at https: / / doi.org / 10.1002 / chem.202300524) but these type of macrocycles have not been investigated for GSIs and chemical strategies for the use and chemical linkages of this type of macrocycles in peptides such as insulin have not been developed

[0006] Accordingly, there is a need for improved glucose binding macrocyles as conjugates for insulin to provide novel GSIs. SUMMARY

[0007] In a first aspect, a molecular conjugate is represented by Formula I, a stereoisomer, a mixture of stereoisomers, or pharmaceutically acceptable salt thereof:

[0008] Formula I

[0009] The moiety PPI can comprises a polypeptide or a polypeptide drug substance comprising an amine. The moiety A can independently for each occurrence be selected from A1 or A2. A1 can be selected from formulas II or III with substituents further described below.

[0010]

[0011] III.

[0012] In a second aspect, the present disclosure includes a pharmaceutical composition comprising at least one compound or a pharmaceutically acceptable salt thereof according to Formula I and pharmaceutically acceptable carrier.

[0013] In a third aspect, the present disclosure includes a compound according to formula I for use as a medicament.

[0014] In a fourth aspect, the present disclosure includes a method of treatment or prevention of diabetes, impaired glucose tolerance, hyperglycemia or metabolic syndrome. The method comprises administering to a subject in need thereof a therapeutically effective amount of a compound according to formula I, or a pharmaceutical composition according to the second aspect.

[0015] In some aspects, a pyridinium-based glucose binding macrocycle is conjugated to insulin along with a binding partner in pursuit of glucose- sensitive insulin (GSI), specifically, a macrocycle at B29 and glucoside at Bl via different linkers. The compounds preferably show good glucose-sensitive bioactivity and weak albumin binding. The given molecules can be useful as meal GSI. Because some glucose binders are known to bind relatively strongly to albumin, and this can lead to delayed absorption from the injection site, a prandial insulin should ideally be absorbed fast. The macrocycles disclosed herein preferably are polar and do not contain multiple carboxylates or boronates, which are known to promote strong albumin binding. DETAILED DESCRIPTION

[0016] As described above, in a first aspect, a molecular conjugate is represented by Formula I, a stereoisomer, a mixture of stereoisomers, or pharmaceutically acceptable salt thereof:

[0017] Formula I The moiety PPI can comprise a polypeptide, a polypeptide drug substance, an insulin, an incretin, such as GLP-1 or GIP, a glucagon. In one embodiment, PPI comprises an amine to which A directly or via B and / or C binds. In one embodiment, the amine can be on an amino acid residue, e.g., lysine or can be the N-terminus of a polypeptide chain in PPI . In another embodiment, PPI comprises a free thiol. Moiety A can independently for each occurrence be selected from A1 or A2. Moiety A is covalently conjugated either directly or via B or C to PPI. In one embodiment, the molecular conjugate comprises at least one A1. Moreover, A1 can be selected from the group of the formulas II and III

[0018]

[0019] III

[0020] In formulas II and III, Zx— is a handle at a vacancy of formulas II or III and conjugated either directly or via B and / or C to PPI. In one embodiment, Zx— is independently for each occasion selected from , wherein p is an integer from 0 to 3, and — denotes a covalent bond either directly, or via B and / or C to PPI.

[0021] Bonds bi, bi, bi, b4, b% and be are independently for each occasion selected from a single bond or a double bond. Substituents R1, R2, R3, R4, R5and R6can independently be selected from hydrogen, carbonyl, C1-C6alkyl, C3-C8 cycloalkyl, aryl, heteroaryl, heterocyclyl, C1-C4alkoxy, C1-C4alkylamino, C1-C4alkyl hydroxy, C1-C4alkyl carboxylate, C1-C4alkyl carbamide. Each of the foregoing substituents R1through R6can optionally have one or more additional substituents selected from C1-C4alkyl, C1-C4alkoxy, C1-C4alkylamino, C1-C4alkylcarboxylate, C1-C4alkylcarbamide, halo, C1-C4haloalkyl, C1-C4haloalkoxy, C1-C4haloalkylamino, amino, cyano, hydroxyl, carboxy, carbamoyl, sulfamoyl, or mercapto.

[0022] In one embodiment, Zx— can be bonded at rings G or H as shown in formulas Ila, lib, III a, or Illb.

[0023]

[0024] Illb

[0025] In one embodiment, Zx— can be part of to R1, R2, R3, R4, R5, or R6and is absent in rings as shown in formulas lib and IIIc.

[0026]

[0027] IIIc, wherein for formulas lie and IIIc, Zx— binds to a vacancy of R1, R2, R3, R4, R5, or R6.

[0028] In one embodiment, R1and R2are linked to form a group of the formula

[0029]

[0030] In the foregoing depictions of rings J, K, and L, the wavy lines " denote the point of attachment to X1, X2, X3, X4, X5, or X6, respectively. In one embodiment, rings J, K and L can independently be selected from arylene, heteroarylene, heterocylylene, cyclylene, heterocycloalkylenylene, and cycloalkylenylene. In one further embodiment, ring substituents R9, R10, and Rn can independently for each occasion be selected from C1-C4alkyl, C1-C4alkoxy, C1-C4alkylamino, C1-C4alkyl hydroxy, C1-C4alkyl carboxylate, C1-C4alkyl carbamide, halo, C1-C4haloalkyl, C1-C4haloalkoxy, C1-C4haloalkylamino, amino, cyano, hydroxyl, carboxyl, carbamoyl, sulfamoyl, and mercapto. The integers c, d, and e can be selected from 0 to 4.

[0031] In one embodiment, rings G and H can independently be selected from arylene or heteroarylene. The ring substituents R7 and Rs can independently for each occasion be selected from C1-C4alkyl, C1-C4alkoxy, C1-C4alkylamino, C1-C4alkylhydroxy, carboxy, C1-C4alkylcarboxylate, C1-C4alkylcarbamide, halo, C1-C4haloalkyl, C1-C4haloalkoxy, C1-C4haloalkylamino, amino, cyano, hydroxyl, carboxy, carbamoyl, sulfamoyl, and mercapto. The integers a and b are selected from 0 to 4. In one embodiment, rings G and H can be a alkyl substituted phenylene, such as 2,3,5,6-tetraalkyl phenylene for macrocyclic moieties according to formula II and 2,4,6-tri-alkyl phenylene for macrocyclic moieties according to formula III.

[0032] The moieties W1, W2, W3, and W4 can independently be selected from CRhR1wherein Rhand R1are independently selected from hydrogen or C1-C3alkyl. The moieties X1, X2, X3, X4, X5and X6can independently be selected from a group of

[0033] In the foregoing groups, the wavy line denotes the point of attachment. In one instant, the left denotes the point of attachment to moiety comprising R1and R2, or the moiety comprising R3and R4, or the moiety comprising R5and R6. In another instant the left denotes the bond between rings J, K, or L, and X1through X6. Accordingly, the right «w denotes the bond between Wxand any of Wi through We. In one instant, Wxis selected from 0 or NH. In one embodiment, the moieties X1, X2, X3, X4, X5and X6can independently be selected from .

[0034] The group of A2 comprises a glycoside, a glycoside derivative, an inositol, a sugar alcohol, or a peptide. For example, for glycosides and derivatives thereof, A2 can include a glucoside, a 6-deoxy-glucoside, a glucuronic acid glucoside, or a xyloside.

[0035] The moiety B of formula I represents a linker and can independently for each occasion be selected from Bi or B2, wherein Bi is a peptide linker and B2 is a chemical linker. The moiety C may be a spacer. The integers of formula I are as follows. Integer a’ is independently for each occasion an integer equal to or greater than 1; b’ is independently for each occasion 0 or an integer; c’ is independently for each occasion 0 or an integer; m’ is independently for each occasion an integer greater than or equal to 1; and n’ is an integer greater than or equal to 1.

[0036] In one embodiment, the molecular conjugate according to the first aspect comprises at least one A1 and at least one A2. According to the present disclosure, molecular conjugates comprising one A1 and one A2 can each be covalently bond either directly or via B or C to PPI. Each of A1 and A2 can be bound at different locations on PPI. For example, if PPI is an insulin, A1 can be covalently bonded either directly or via B or C to an amino acid near or at the C- terminus of the B-chain while A2 can be covalently bonded either directly or via another B or C to an amino acid near or at the N-terminus of the B-chain or the N-terminus of the A-chain of the insulin. Likewise, the reverse configuration is contemplated within the scope of the present disclosure, where A2 can be covalently bonded either directly or via B or C to an amino acid near or at the C -terminus of the B -chain while A1 can be covalently bonded either directly or via another B or C to an amino acid near or at the N-terminus of the B-chain or the N-terminus of the A-chain of the insulin

[0037] In one further embodiment, the molecular conjugate according to the first aspect of the disclosure comprises at least one of rings J, K, or L and each ring independently for each occasion can include a ring according to the following structure:

[0038] In the foregoing structure, Xaindependently for each occasion can be selected from CH, CRa, N, or N+Ra. Substituent Racan independently for each occasion be selected from Zx— , C1-C4alkylcarboxylate, C1-C4alkylcarbamide, C1-C4alkylamine, or C1-C4alkylhydroxy. In one Racan be selected from C1-C4alkyl, wherein integer q is 1, 2, or 3 and — denotes the covalent bond either directly, or via B and / or C to PPI.

[0039] In one further embodiment, rings J, K, or L can independently for each occasion comprise

[0040] Yet, in one particular embodiment, the molecular conjugate according to first aspect includes at least one of rings J, K, or L independently for each occasion which comprises

[0041] In the foregoing pyridinium moiety, the wavy lines denote the point of attachment to one of X1, X2, or X3, X4, or X5, X6, respectively. The dashed line represents the bond directly to or via B or C to PPI.

[0042] In yet another embodiment, the molecular conjugate according to formula I includes roof and floor rings G and H for A1 in formula II. Rings G and H of formula II can be

[0043] As for rings G and H of formula III, they can be selected from

[0044] In the foregoing structures, Rb can be any of substituents R7 and Rs. For example, Rb can independently for each occasion b selected from hydrogen, C1-C4alkyl, C1-C4haloalkyl, C1-C4hydroxyalkyl, C1-C4amino-alkyl, carboxy or C1-C4alkylcarboxylate, or C1-C4alkylcarboxamide, and Wu, Wu’, and Wu” are selected from Wj, W2, W3, W4, W5, or We of formula II and III, respectively.

[0045] In one embodiment, Ai is selected from

[0046]

[0047] The counterion X|,®is a pharmaceutically acceptable counterion. For example, Xb® can be chloride. The dashed line — denotes the covalent bond to B, C, or PPI.

[0048] Addressing the moiety A2, this moiety includes glycosides or glycoside derivatives. The glycoside can be D- or L-glycosides and D- or L-glycoside derivatives. In embodiments, moiety A2 can include alpha-D-glucopyranoside, alpha-D-mannopyranoside, alpha-D-galactopyranoside, beta-D-glucopyranoside, beta-D-mannopyranoside, beta-D-galactopyranoside, alpha-L- glucopyranoside, alpha-L-mannopyranoside, alpha- L-galactopyranoside beta-L-glucopyranoside, beta-L-mannopyranoside, beta-L-galactopyranoside, and derivatives thereof. Derivatives can include D- or L-forms of 6-deoxy glycosides, 6-carboxy glycoside, or 6-deoxy-6-desmethyl glycosides. In one embodiment, A2 can be selected from wherein — denotes a covalent bond to B, C, or directly PPI. In another embodiment A2 can be a peptide. For example, A2 can be peptide that binds to Ai in similar manner as a glycoside, i.e., an Ai -binding peptide.

[0049] In some embodiments, the A2 is a peptide that binds to Ai in a glucose-dependent manner. For example, the affinity of the A2 as a peptide binding to Ai is affected by the concentration of glucose.

[0050] In another embodiment, the molecular conjugate according to the first aspect includes moiety B, wherein B can be selected from Bi or B2. Bi can be absent or independently for each occasion selected from the group of an amino acid and a peptide comprising two to eight amino acids. Likewise, B2 can be absent or independently selected from

[0051] In the foregoing structures, Sw represents CH2— - or (C=O)— -, wherein — - is a covalent bond to moiety C or PPI of formula I. Each Sv is independently selected from — 0—*, NH— *, CH2— *, and (C=0)— * and each Svi is N— *, wherein — * is a covalent bond to A1 or A2, integer r is selected from 0 to 8, and integer t is selected from 0 to 6. In one embodiment, B2 can be selected from wherein integer u is selected from 1 to 6, — denotes a covalent bond to C, or directly PPI, and *- - denotes a covalent bond to A.

[0052] In another embodiment, the molecular conjugate according to the first aspect includes moiety C. Moiety C can be absent or independently selected for each occasion from the group of an amino acid or a peptide comprising two to eight amino acids.

[0053] In another embodiment, the molecular conjugate according to the first aspect includes moiety PPI. Group PPI can include a polypeptide human hormone, an endocrine hormone, insulin, human insulin, glucagon, amylin, relaxin, GLP-1, oxyntomodulin, somatostatin, gastric inhibitory polypeptide, glucose-dependent insulinotropic polypeptide, a hybrid peptide comprising sequences from two or more human polypeptide hormones, or an analogue of any thereof. In one embodiment, PPI is human insulin. In one particular embodiment, PPI is insulin and has an A-chain having the amino acid sequence GIVEQCCTSICSLYQLENYCN, and a B- chain having the amino acid sequence FVNQHLCGSHLVEALYLVCGERGFFYTPK. Moreover, in one embodiment PPI is human insulin comprising at least one of the following mutations: A14E, A21G, B3E, B16H, B25H, B28D, B28K, B29P, or desB30.

[0054] Having described the moieties of formula I above, the following are examples of the molecular conjugates:

[0055]

[0056]

[0057] In a second aspect, the present disclosure includes a pharmaceutical composition comprising at least one compound or a pharmaceutically acceptable salt thereof according to Formula I and pharmaceutically acceptable carrier. In a third aspect, the present disclosure includes a compound according to formula I for use as a medicament.

[0058] In a fourth aspect, the present disclosure includes a method of treatment or prevention of diabetes, impaired glucose tolerance, hyperglycemia or metabolic syndrome. The method comprises administering to a subject in need thereof a therapeutically effective amount of a compound according to formula I, or a pharmaceutical composition according to the second aspect.

[0059] Without limiting the full scope of the present disclosure, the following item list describe embodiments of the present disclosure:

[0060] Item 1. A molecular conjugate represented by Formula I, a stereoisomer, a mixture of stereoisomers, or pharmaceutically acceptable salt thereof:

[0061] Formula I wherein

[0062] PPI comprises a polypeptide drug substance,

[0063] A is independently for each occurrence selected from A1 or A2 and is covalently conjugated either directly or via B and / or C to PPI, wherein the molecular conjugate comprises at least one A1 and the A1 is selected from the group of the formulas II and III

[0064]

[0065] III wherein Zx— is a handle at a vacancy of formulas II or III and conjugated either directly or via B and / or C to PPI and Zx— is selected from , , wherein p is an integer from 0 to 3, — denotes a covalent bond either directly, or via B and / or C to PPI; bi, b2, ba, b4, bs, and be are independently for each occasion selected from a single bond or a double bond. R1, R2, R3, R4, R5and R6can independently be selected from hydrogen, carbonyl, C1-C6alkyl, C3-C8 cycloalkyl, aryl, heteroaryl, heterocyclyl, C1-C4alkoxy, C1-C4alkylamino, C1-C4alkylhydroxy, C1-C4alkylcarboxylate, C1-C4alkylcarbamide, which is optionally substituted by one or more substituents selected from C1-C4alkyl, C1-C4alkoxy, C1-C4alkylamino, C1-C4alkylcarboxylate, C1-C4alkylcarbamide, halo, C1-C4haloalkyl, C1-C4haloalkoxy, C1-C4haloalkylamino, amino, cyano, hydroxyl, carboxy, carbamoyl, sulfamoyl, and mercapto; or R1and R2are linked to form a group of the formula or R3and R4are linked to form a group of the formula or R5and R6are linked to form a group of the formula wherein denotes the point of attachment to X1, X2, X3, X4, X5, and X6, respectively;

[0066] Rings J, K and L are independently selected from arylene, heteroarylene, heterocylylene, cyclylene, heterocycloalkylenylene, and cycloalkylenylene,

[0067] R9, R10and Rn are independently for each occasion selected from C1-C4alkyl, C1- C4alkoxy, C1-C4alkylamino, C1-C4alkylhydroxy, C1-C4alkylcarboxylate, carboxylate, C1-C4alkylcarbamide, halo, C1-C4haloalkyl, C1-C4haloalkoxy, C1-C4haloalkylamino, amino, cyano, hydroxyl, carboxyl, carbamoyl, sulfamoyl, and mercapto, integers c, d, and e are selected from 0 to 4;

[0068] Rings G and H are independently selected from arylene or heteroarylene,

[0069] R7 and Rs are independently for each occasion selected from C1-C4alkyl, C1-C4alkoxy, C1-C4hydroxy alkyl, C1-C4amino-alkyl, C1-C4alkylhydroxy, C1-C4alkylcarbamide, C1-C4alkylcarboxylate, C1-C4alkylamino, halo, C1-C4haloalkyl, C1-C4haloalkoxy, C1-C4haloalkylamino, amino, cyano, hydroxyl, carboxy, carbamoyl, sulfamoyl, and mercapto, integers a and b are selected from 0 to 4;

[0070] Wi, W2, W3, and W4 are independently selected from C RhRiwherein Rhand Riare independently selected from hydrogen or C1-C3alkyl X1, X2, X3, and X4are independently selected from a group of

[0071] denotes the point of attachment to the rest of the macrocycle, Wxis selected from 0 or NH; wherein A2 comprises a peptide, a polyol containing moiety, a glycoside, a glycoside derivative, an inositol, or a sugar alcohol; wherein B is independently for each occasion is a spacer selected from Bi or B2, wherein Bi is a peptidic linker and B2 is a chemical linker; wherein C is a spacer; and a’ is independently for each occasion an integer equal to or greater than 1 ; b’ is independently for each occasion 0 or an integer; c’ is independently for each occasion 0 or an integer; m’ is independently for each occasion an integer greater than or equal to 1 ; n’ is an integer greater than or equal to 1.

[0072] Item 2. The molecular conjugate according to item 1 , wherein Ai is selected from

[0073]

[0074] lie

[0075] IIIc, wherein for formulas lie and IIIc, Zx— binds to a vacancy of R1, R2, R3, R4, Rs, or R6.

[0076] Item 3. The molecular conjugate according to any one of the preceding items further comprising at least one A2.

[0077] Item 4. The molecular conjugate according to any one of the preceding items, wherein rings J,

[0078] K, or L independently for each occasion comprises wherein Xaindependently for each occasion is selected from CH, CRa, N, or N+Ra; wherein Raindependently for each occasion is selected from C1-C4alkyl, 0 , wherein integer q is 1 , 2, or 3 and — denotes the covalent bond either directly, or via B and / or C to PPI . Item 5. The molecular conjugate according to item 4, wherein rings J, K, or L independently for each occasion comprises

[0079] Item 6. The molecular conjugate according to any one of the preceding items, wherein at least one of rings J, K, or L independently for each occasion comprises

[0080] Item 7. The molecular conjugate according to any one of the preceding items, wherein rings

[0081] G and H of formula II are selected from rings G and H of formula III are selected from haloalkyl, C1-C4hydroxyalkyl, C1-C4amino-alkyl, carboxy, or C1-C4alkylcarboxylate, or C1-C4 alkylcarboxamide, and Wu, WU’, and WU” are selected from Wi, W2, W3, W4, W5, or We, respectively.

[0082] Item 8. The molecular conjugate according to any one of the preceding items, wherein Ai is selected from wherein X6Qis a pharmaceutically acceptable counterion and — denotes the covalent bond to B, C, or PPI.

[0083] Item 9. The molecular conjugate according to any one of the preceding items, wherein A2 is a D-glycoside, a D-glycoside derivative, an L- glycoside, an L- glycoside derivative, or an Ai- binding peptide.

[0084] Item 10. The molecular conjugate according to any one of the preceding items, wherein A2 is selected from wherein — denotes a covalent bond to B, C, or directly PPI. Item 11. The molecular conjugate according to any one of the preceding items, wherein Bi is absent or independently for each occasion selected from the group of an amino acid and a peptide comprising two to eight amino acids.

[0085] Item 12. The molecular conjugate according to any one of the preceding items, wherein Bi is absent or independently selected from

[0086] Sw represents CH2— - or (C=O)— -, wherein — - is a covalent bond to C or PPI; and each Sv is independently selected from 0— *, NH— *, CH2— *, and (C=0)— * and each Svi is N— *, wherein — * is a covalent bond to A1 or A2, integer r is selected from 0 to 8, and integer t is selected from 0 to 6.

[0087] Item 13. The molecular conjugate according to any one of the preceding items, wherein B2 is selected from wherein integer u is selected from 1 to 6, — denotes a covalent bond to C, or directly PPI, and *- - denotes a covalent bond to A.

[0088] Item 14. The molecular conjugate according to any one of the preceding items, wherein C is absent or independently selected for each occasion from the group of an amino acid or a peptide comprising two to eight amino acids.

[0089] Item 15. The molecular conjugate according to any one of the preceding items, wherein PPI comprises a polypeptide human hormone, an endocrine hormone, insulin, human insulin, glucagon, amylin, relaxin, GLP-1, oxyntomodulin, somatostatin, gastric inhibitory polypeptide, glucose-dependent insulinotropic polypeptide, a hybrid peptide comprising sequences from two or more human polypeptide hormones, or an analogue of any thereof. Item 16. The molecular conjugate according to any one of the preceding items, wherein PPI is human insulin or an analogue thereof.

[0090] Item 17. The molecular conjugate of item 16, wherein the insulin has 1, 2, 3, 4, 5, or 6 mutations, deletions or amino acid additions.

[0091] Item 18. The molecular conjugate according to item 16, wherein the human insulin has an A- chain having the amino acid sequence GIVEQCCTSICSLYQLENYCN, and a B-chain having the amino acid sequence FVNQHLCGSHLVEALYLVCGERGFFYTPK.

[0092] Item 19. The molecular conjugate according to any one of the preceding items, wherein PPI is human insulin comprising at least one of the following mutations: A14E, A21G, B3E, B16H, B25H, B28D, B28K, B29P, or desB30. Item 20. The molecular conjugate according to any one of the preceding items selected from

[0093]

[0094]

[0095]

[0096]

[0097] Item 21. A pharmaceutical composition comprising at least one compound or a pharmaceutically acceptable salt thereof according to any one of items 1-20 and a pharmaceutically acceptable carrier.

[0098] Item 22. A compound according to any one of items 1-20, for use as a medicament. Item 23. A method of treatment or prevention of diabetes, impaired glucose tolerance, hyperglycemia or metabolic syndrome, wherein the method comprises administering to a subject in need thereof a therapeutically effective amount of the compound of any one of items 1-20, or a pharmaceutical composition according to item 21.

Claims

WHAT IS CLAIMED IS:

1. A molecular conjugate represented by Formula I, a stereoisomer, a mixture of stereoisomers, or pharmaceutically acceptable salt thereof:Formula I whereinPPI comprises a polypeptide drug substance,A is independently for each occurrence selected from A1 or A2 and is covalently conjugated either directly or via B and / or C to PPI, wherein the molecular conjugate comprises at least one A1 and the A1 is selected from the group of the formulas II and IIIwherein Zs— is a handle at a vacancy of formulas II or III and conjugated either directly or via B and / or C to PPI and Zx— is selected from, , wherein p is an integer from 0 to 3, — denotes a covalent bond either directly, or via B and / or C to PPI ; bi, bi, hi, b4, bs, and be are independently for each occasion selected from a single bond or a double bond. R1, Ri, R3, R4, Rs and R6can independently be selected from hydrogen, carbonyl, Ci-Ce alkyl, C3-C8 cycloalkyl, aryl, heteroaryl, heterocyclyl, C1-C4alkoxy, C1-C4alkylamino, C1-C4alkylhydroxy, C1-C4alkylcarboxylate, C1-C4alkylcarbamide, which is optionally substituted by one or more substituents selected from C1-C4alkyl, C1-C4alkoxy, C1-C4alkylamino, C1-C4alkylcarboxylate, C1-C4alkylcarbamide, halo, C1-C4haloalkyl, Ci- C4 haloalkoxy, C1-C4haloalkylamino, amino, cyano, hydroxyl, carboxy, carbamoyl, sulfamoyl, and mercapto; or R1and R2are linked to form a group of the formulaor R3and R4are linked to form a group of the formulawherein■~w denotes the point of attachment to X1, X2, X3, X4, X5, and X6, respectively;Rings J, K and L are independently selected from arylene, heteroarylene, heterocylylene, cyclylene, heterocycloalkylenylene, and cycloalkylenylene,R9, R1o and Rn are independently for each occasion selected from C1-C4alkyl, C1-C4alkoxy, C1-C4alkylamino, C1-C4alkylhydroxy, C1-C4alkylcarboxylate, carboxylate, C1-C4alkylcarbamide, halo, C1-C4haloalkyl, C1-C4haloalkoxy, C1-C4haloalkylamino, amino, cyano, hydroxyl, carboxyl, carbamoyl, sulfamoyl, and mercapto, integers c, d, and e are selected from 0 to 4;Rings G and H are independently selected from arylene or heteroarylene,R7 and Rs are independently for each occasion selected from C1-C4alkyl, C1-C4alkoxy, Ci- C4 hydroxyalkyl, C1-C4amino-alkyl, C1-C4alkylhydroxy, C1-C4alkylcarbamide,C1-C4alkylcarboxylate, C1-C4alkylamino, halo, C1-C4haloalkyl, C1-C4haloalkoxy, C1-C4haloalkylamino, amino, cyano, hydroxyl, carboxy, carbamoyl, sulfamoyl, and mercapto, integers a and b are selected from 0 to 4;Wi, W2, W3, and W4 are independently selected from CRhR1wherein Rhand R1are independently selected from hydrogen or C1-C3 alkyl X1, X2, X3, and X4are independently selected from a group ofdenotes the point of attachment to the rest of the macrocycle,Wx is selected from 0 or NH; wherein A 2 comprises a peptide, a polyol containing moiety, a glycoside, a glycoside derivative, an inositol, or a sugar alcohol; wherein B is independently for each occasion is a spacer selected from Bi or Bi, wherein Bi is a peptidic linker and B2 is a chemical linker; wherein C is a spacer; and a’ is independently for each occasion an integer equal to or greater than 1 ; b’ is independently for each occasion 0 or an integer; c’ is independently for each occasion 0 or an integer; m’ is independently for each occasion an integer greater than or equal to 1; n’ is an integer greater than or equal to 1.

2. The molecular conjugate according to claim 1, wherein Ai is selected fromliblieIIIc, wherein for formulas lie and IIIc, Zx— binds to a vacancy of R1, R2, R3, R4, Rs, or R6.

3. The molecular conjugate according to claim 1 further comprising at least one A2.

4. The molecular conjugate according to claim 1 , wherein rings J, K, or L independently for each occasion compriseswherein Xaindependently for each occasion is selected from CH, CRa, N, or N+Ra;% wherein Raindependently for each occasion is selected from C1-C4alkyl,0 0Hwherein integer q is 1, 2, or 3 and — denotes the covalent bond either directly, or via B and / or C to PPI.

5. The molecular conjugate according to claim 1, wherein rings J, K, or L independently for each occasion comprises6. The molecular conjugate according to claim 1, wherein at least one of rings J, K, or L independently for each occasion comprises7. The molecular conjugate according to claim 1, wherein rings G and H of formula II are selected fromrings G and H of formula III are selected from, whereinRb independently for each occasion is selected from hydrogen, C1-C4alkyl, C1-C4haloalkyl, C1-C4hydroxyalkyl, C1-C4amino-alkyl, carboxy, or C1-C4alkylcarboxylate, or C1-C4alkylcarboxamide, and Wu, Wu’, and Wu” are selected from Wi, W2, W3, W4, W5, or We, respectively.

8. The molecular conjugate according to claim 1, wherein Ai is selected fromwherein Xb® is a pharmaceutically acceptable counterion and — denotes the covalent bond to B, C, or PPI.

9. The molecular conjugate according to claim 1 wherein A2 is a D-glycoside, a D-glycoside derivative, an L-glycoside, an L-glycoside derivative, or an Ai -binding peptide.

10. The molecular conjugate according to claim 1 wherein A2 is selected fromwherein — denotes a covalent bond to B, C, or directly PPI.

11. The molecular conjugate according to claim 1 wherein Bi is absent or independently for each occasion selected from the group of an amino acid and a peptide comprising two to eight amino acids.

12. The molecular conjugate according to claims 1 wherein B2 is absent or independently selected fromSw represents CH2— - or (C=0)— wherein — - is a covalent bond to C or PPI ; and each Sv is independently selected from —0—*, NH— *, CH2— *, and (C=0)— * and each Svi is N— *, wherein — * is a covalent bond to A1 or A2, integer r is selected from 0 to 8, and integer t is selected from 0 to 6.

13. The molecular conjugate according to claim 1 wherein B2 is selected fromwherein integer u is selected from 1 to 6, — denotes a covalent bond to C, or directly PPI, and *— denotes a covalent bond to A.

14. The molecular conjugate according to claim 1 wherein C is absent or independently selected for each occasion from the group of an amino acid or a peptide comprising two to eight amino acids.

15. The molecular conjugate according to claim 1 wherein PPI comprises a polypeptide human hormone, an endocrine hormone, insulin, human insulin, glucagon, amylin, relaxin, GLP-1, oxyntomodulin, somatostatin, gastric inhibitory polypeptide, glucose-dependent insulinotropic polypeptide, a hybrid peptide comprising sequences from two or more human polypeptide hormones, or an analogue of any thereof.

16. The molecular conjugate according to claim 1, wherein PPI is human insulin or an analogue thereof.

17. The molecular conjugate of claim 15 wherein the insulin has 1, 2, 3, 4, 5, or 6 mutations, deletions or amino acid additions.

18. The molecular conjugate according to claim 16, wherein the human insulin has an A-chain having the amino acid sequence GIVEQCCTSICSLYQLENYCN, and a B-chain having the amino acid sequence FVNQHLCGSHLVEALYLVCGERGFFYTPK.

19. The molecular conjugate according to claim 1, wherein PPlis human insulin comprising at least one of the following mutations: A14E, A21G, B3E, B16H, B25H, B28D, B28K, B29P, or desB30.

20. The molecular conjugate according to claim 1 selected from21. A pharmaceutical composition comprising at least one compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-20 and a pharmaceutically acceptable carrier.

22. A compound according to any one of claims 1-20, for use as a medicament.

23. A method of treatment or prevention of diabetes, impaired glucose tolerance, hyperglycemia or metabolic syndrome, wherein the method comprises administering to a subject in need5 thereof a therapeutically effective amount of the compound of any one of claims 1-20, or a pharmaceutical composition according to claim 21.

Images