Peptides for the treatment and prevention of diabetes and related disorders

Abstract

To provide a pharmaceutical composition for treating diabetes mellitus and related disorders.SOLUTION: A pharmaceutical composition includes peptide, and is used for treating diabetes mellitus and related disorders selected from the group consisting of type I diabetes mellitus, type II diabetes mellitus, insulin resistance, diabetic retinopathy, diabetic neuropathy, diabetic nephropathy, hyperglycemia, obesity, hyperinsulinemia and Bardet-Biedl syndrome, where the peptide can reduce an expression of FATP2 in an adipose tissue, does not simultaneously contain a single methionine, a single proline and a single arginine, adopts a secondary structure being helix, preferably alpha-helix, has the length of 5 to 60 amino acid, and includes a specific sequence.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to the field of medicine, and more particularly to the treatment of diabetes and related disorders. [Background technology]

[0002] Diabetes mellitus, or diabetes, is a metabolic disease in which patients suffer from hyperglycemia because the pancreas does not produce enough insulin or because cells do not respond to the insulin that is secreted.

[0003] There are three main types of diabetes. - Type 1 is caused by the body not producing insulin, and currently patients must inject insulin or wear an insulin pump. - Type 2 is caused by insulin resistance, a condition in which cells are unable to use insulin properly. - The third type is called gestational diabetes, which occurs in women who are pregnant.

[0004] The incidence of type 2 diabetes has increased significantly since 1960, in parallel with obesity. As of 2010, there are approximately 285 million people with type 2 diabetes, compared to approximately 30 million in 1985. Long-term complications of hyperglycemia include heart disease, stroke, diabetic retinopathy, chronic kidney failure requiring dialysis, and poor circulation in the hands and feet leading to amputation. Nonketotic hyperosmolar coma may also develop.

[0005] It has been reported that hyperglycemia is involved in the onset and progressive functional disorders of diabetes mellitus, i.e., the glucose toxicity hypothesis. That is, chronic hyperglycemia reduces insulin secretion and further reduces insulin sensitivity, resulting in an increase in blood glucose concentration and self-aggravation of diabetes mellitus. Therefore, by treating hyperglycemia, the aforementioned self-aggravation cycle can be broken, making it possible to prevent or treat diabetes mellitus.

[0006] Unfortunately, existing treatments are unable to restore normal blood glucose levels in the long term because beta-cell function declines over time, and there is currently no single drug that can reverse all aspects of the disease.

[0007] Due to the progressive nature of type 2 diabetes, many patients will eventually need to use oral hypoglycemic agents in combination with possible insulin and / or exenatide injections. Antidiabetic drugs have been developed to combat the main mechanisms involved in type 2 diabetes: insulin resistance (biguanides, thiazolidinediones) and insulin secretion (sulfonylureas, glinides, dipeptidyl peptidase-4 inhibitors, glucagon-like peptide 1 receptor agonists). New drugs have been developed to slow the absorption of glucose in the gastrointestinal tract, promote weight loss, and promote renal glucose excretion. However, many of these drugs have been found to have adverse side effects, such as weight gain, peripheral edema, and congestive heart failure, and there is a major problem of diminishing efficacy with long-term use. Thus, despite the increasing number of therapeutic options available to control blood glucose levels, alternative and improved drugs are needed to treat diabetes and its related diseases. [Prior art documents] [Patent documents]

[0008] [Patent Document 1] WO10033617 [Patent Document 2] WO10011313 [Patent Document 3] WO2005 / 117830 [Patent Document 4] WO2006 / 075124 [Patent Document 5] WO2015 / 114062 [Patent Document 6] WO2004 / 103995 [Patent Document 7] WO03 / 043985 [Non-patent literature]

[0009] [Non-Patent Document 1] Frishman D., Argos P., Proteins, Vol. 23, No. 4, 1995, pp. 566-579 [Non-Patent Document 2] Richards FM, Kundrot CE, Proteins, vol. 3, no. 2, 1988, pp. 71-84 [Non-Patent Document 3] Touw et al., Nucleic Acids Research 2015, 43:D364~D368 [Non-Patent Document 4] Kabsch and Sander, Biopolymers., 1983, 22, 2577-2637 [Non-Patent Document 5] Verdine and Hilinski (2012, Methods Enzymol, 503, 3~33) [Non-Patent Document 6] Matsushita and Matsui, (2005), J Mol Med 83, 324-328 [Non-Patent Document 7] Vives et al., Biochimic et Biophysica Acta, 2008, 1786, 126–138 [Non-Patent Document 8] Expert Opin Investig Drugs 2003, 12(4):623~633 [Non-Patent Document 9] Lippincott Williams & Wilkins, 2000 and Encyclopedia of Pharmaceutical Technology, edited by J. Swarbrick and JC Boylan, 1988-1999, Marcel Dekker, New York. Summary of the Invention [Problem to be solved by the invention]

[0010] Surprisingly, the present inventors provide a peptide and its derivatives derived from the kinase domain of PKCα that specifically improves glucose tolerance in diet-induced obese mice. The peptide can reduce the expression of Solute Carrier Family 27 Member 2 (SLC27A2), commonly known as FATP2 (Fatty acid transport protein 2), in adipose tissue. The peptide can reduce glycated albumin in plasma of animal models, which is a well-known biomarker for diabetes. [Means for solving the problem]

[0011] Accordingly, the present invention provides a peptide for use in the treatment of diabetes and related disorders, comprising: - the peptide is capable of specifically reducing the expression of FATP2 in adipose tissue, in particular in mammalian, in particular human, adipose tissue, the peptide does not simultaneously contain one methionine, one proline and one arginine; - the peptide adopts a secondary structure which is a helix, preferably an alpha-helical - the peptide comprises, consists essentially of, or consists of a sequence derived from a segment of at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 25 consecutive residues of the kinase domain of PKC (protein kinase C), or a segment of 5 to 40 consecutive residues of the kinase domain of PKC (protein kinase C), the peptide has a length of 5 to 80 amino acids, or 5 to 60 amino acids, or 5 to 40 amino acids, - the peptide sequence relates to a peptide which may contain within said sequence of a segment of the kinase domain of PKC 1, 2, 3, 4 or 5 modifications of amino acids selected from substitutions, deletions, additions and combinations thereof.

[0012] Preferably, the peptides are modified by a chemical cross-linking process such as stapling.

[0013] Preferably, the peptide has a length of at least 5 amino acids and less than 40 amino acids, preferably at least 5 amino acids and less than 30 amino acids, more preferably at least 5 amino acids and less than 25 amino acids.

[0014] Preferably, the peptide is capable of reducing or preventing the interaction between ALMS1 and αPKC.

[0015] Optionally, the peptide sequence has the following sequence: VECTMVEKRVLA (SEQ ID NO: 3), optionally with 1, 2, 3, 4, or 5 amino acid modifications selected from substitution, deletion, addition, and combinations thereof, more preferably with 1, 2, or 3 amino acid modifications selected from substitution, deletion, addition, and combinations thereof; VECTXVEKRVLA (SEQ ID NO: 9), optionally with 1, 2, or 3 amino acid modifications selected from substitution, deletion, addition, and combinations thereof, more preferably with 1, 2, or 3 amino acid modifications selected from substitution, deletion, addition, and combinations thereof; VECTMVEKXVLA (SEQ ID NO: 10), optionally with 1, 2, 3, 4, or 5 amino acid modifications selected from substitution, deletion, addition, and combinations thereof, more preferably with 1, 2, or 3 amino acid modifications selected from substitution, deletion, addition, and combinations thereof; VECTXVEKXVLA (SEQ ID NO:11), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof; LMYHIQQV (SEQ ID NO:4), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof; LXYHIQQV (SEQ ID NO:12), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof; LDN; SVDWWAYGVLLYEMLA (SEQ ID NO: 6), optionally having 1, 2, 3, 4, or 5 amino acid modifications selected from substitution, deletion, addition, and combinations thereof, more preferably having 1, 2, or 3 amino acid modifications selected from substitution, deletion, addition, and combinations thereof; SVDWWAYGVLLYEXLA (SEQ ID NO: 13), optionally having 1, 2, 3, 4, or 5 amino acid modifications selected from substitution, deletion, addition, and combinations thereof, more preferably having 1, 2, or 3 amino acid modifications selected from substitution, deletion, addition, and combinations thereof;SVXWWAYGLLYEMLA (SEQ ID NO:52), optionally comprising 1 to 5 modifications of amino acids selected from substitution, deletion, addition, and combinations thereof; EDEDELFQSIME (SEQ ID NO:7), optionally having 1, 2, 3, 4, or 5 modifications of amino acids selected from substitution, deletion, addition, and combinations thereof, more preferably having 1, 2, or 3 modifications of amino acids selected from substitution, deletion, addition, and combinations thereof; EDEDELFQSIXE (SEQ ID NO:14), optionally having 1, 2, 3, 4, or 5 modifications of amino acids selected from substitution, deletion, addition, and combinations thereof, more preferably having 1, 2, or 3 modifications of amino acids selected from substitution, deletion, addition, and combinations thereof; GERDVRE (SEQ ID NO:8), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof; GEXDVRE (SEQ ID NO:15), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof; GERDVXE (SEQ ID NO:16), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof; GEXDVXE (SEQ ID NO:17), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof; LDN; AFF; PDY; XDY; PEII (SEQ ID NO:5); XEII (SEQ ID NO:18); PAK; XAK, where X is any amino acid except M, P, and R;

[0016] Optionally, the peptide sequence has the following sequence: VECTMVEKRVLA (SEQ ID NO:3), optionally with one, two, or three amino acid modifications selected from substitutions, deletions, additions, and combinations thereof; VECTXVEKRVLA (SEQ ID NO:9), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof; VECTMVEKXVLA (SEQ ID NO:10), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof; VECTXVEKXVLA (SEQ ID NO:11), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof; LMYHIQQV (SEQ ID NO:4), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof; LXYHIQQV (SEQ ID NO:12), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof; SVDWWAYGVLLYEMLA (SEQ ID NO:13), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof. 6); SVDWWAYGVLLYEXLA (SEQ ID NO: 13), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof; EDEDELFQSIME (SEQ ID NO: 7), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof; EDEDELFQSIXE (SEQ ID NO: 14), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof; GERDVRE (SEQ ID NO: 8), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof; GEXDVRE (SEQ ID NO: 15), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof; GERDVXE (SEQ ID NO: 16), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof;comprising, consisting essentially of, or consisting of at least one of GEXDVXE (SEQ ID NO: 17), optionally with one, two, or three amino acid modifications selected from substitutions, deletions, additions, and combinations thereof, where X is any amino acid except M, P, and R;

[0017] Optionally, the peptide sequence is the following sequence: a) VECTXVEKXVLALLDKXXFLTQLHS (SEQ ID NO: 20), optionally with 1, 2, 3, 4 or 5 amino acid modifications selected from substitution, deletion, addition and combinations thereof, more preferably with 1, 2 or 3 amino acid modifications selected from substitution, deletion, addition and combinations thereof, where X is any amino acid except M, P and R, preferably an amino acid that favors an alpha helical secondary structure, more preferably an amino acid selected from the group consisting of A, D, N, C, G, Q, E, H, L, K, F, S, W and Y, even more preferably A, D, N, G, Q, E, H, L, K, F, S, W and Y; b) VECTMVEKRVLALLDKXXFLTQLHS (SEQ ID NO: 21), optionally with 1, 2, 3, 4 or 5 amino acid modifications selected from substitution, deletion, addition and combinations thereof, more preferably with 1, 2 or 3 amino acid modifications selected from substitution, deletion, addition and combinations thereof, where X is any amino acid except M, P and R, preferably an amino acid that favors an alpha helical secondary structure, more preferably an amino acid selected from the group consisting of A, D, N, C, G, Q, E, H, L, K, F, S, W and Y, even more preferably A, D, N, G, Q, E, H, L, K, F, S, W and Y; c) VECTXVEKRVLALLDKPPFLTQLHS (SEQ ID NO: 22), optionally with 1, 2, 3, 4, or 5 amino acid modifications selected from substitution, deletion, addition, and combinations thereof, more preferably with 1, 2, or 3 amino acid modifications selected from substitution, deletion, addition, and combinations thereof, where X is any amino acid except M, P, and R, preferably an amino acid that favors an alpha-helical secondary structure, more preferably an amino acid selected from the group consisting of A, D, N, C, G, Q, E, H, L, K, F, S, W, and Y, even more preferably an amino acid selected from the group consisting of A, D, N, G, Q, E, H, L, K, F, S, W, and Y; d) VECTMVEKXVLALLDKPPFLTQLHS (SEQ ID NO: 23), optionally with 1, 2, 3, 4 or 5 amino acid modifications selected from substitution, deletion, addition and combinations thereof, more preferably with 1, 2 or 3 amino acid modifications selected from substitution, deletion, addition and combinations thereof, where X is any amino acid except M, P and R, preferably an amino acid that favors an alpha helical secondary structure, more preferably an amino acid selected from the group consisting of A, D, N, C, G, Q, E, H, L, K, F, S, W and Y, even more preferably A, D, N, G, Q, E, H, L, K, F, S, W and Y; and a sequence of any segment of at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 25 consecutive residues from any of the sequences a) to d). Comprising, consisting essentially of, or consisting of at least one of:

[0018] Optionally, the peptide sequence is the following sequence:

[0019] [ka]

[0020] comprising, consisting essentially of, or consisting of at least one of In the formula, the bold and underlined

[0021] [ka]

[0022] the residues of which comprise the staple and are any amino acid derivative suitable for stapling; X is any amino acid except M, P, and R; The sequence optionally has 1, 2, 3, 4, or 5 amino acid modifications selected from substitution, deletion, addition, and combinations thereof, and more preferably 1, 2, or 3 amino acid modifications selected from substitution, deletion, addition, and combinations thereof.

[0023] Preferably, the PKC is selected from the group consisting of alpha PKC (αPKC), beta PKC (βPKC), including βI and βII PKC, delta PKC, theta PKC, eta PKC and epsilon PKC. More preferably, the PKC is an αPKC of SEQ ID NO:1.

[0024] In certain embodiments, the peptide sequence is

[0025] [ka]

[0026] comprising, consisting essentially of, or consisting of, During the ceremony,

[0027] [ka]

[0028] have staples, preferably 2-(7-octenyl)arginine and 2-(4-pentenyl)serine, respectively; The sequence optionally has 1, 2, 3, 4, or 5 amino acid modifications selected from substitution, deletion, addition, and combinations thereof, and more preferably 1, 2, or 3 amino acid modifications selected from substitution, deletion, addition, and combinations thereof.

[0029] The present invention also relates to a pharmaceutical composition comprising a peptide according to the present disclosure for use in the treatment of diabetes and related disorders, and further to the use of a peptide according to the present disclosure for the manufacture of a medicament for the treatment of diabetes and related disorders.

[0030] Optionally, the diabetes and related disorders are selected from the group consisting of type I diabetes, type II diabetes, insulin resistance, diabetic retinopathy, diabetic neuropathy, diabetic nephropathy, hyperglycemia, obesity, hyperinsulinemia and Bardet-Biedl syndrome. Optionally, the peptide is used in combination with one or more additional agents, preferably selected from the group consisting of antidiabetic agents, lipid lowering agents, antiobesity agents, antihypertensive agents, antilipidemic agents, antiinflammatory agents, and agonists of peroxisome proliferator-activated receptors. [Brief description of the drawings]

[0031] [Figure 1] Figure 1. PKC kinase activity in human adipocytes induced by peptides derived from alpha PKC. Human primary adipocytes were cultured overnight without insulin, and the next day, 25 μg of peptide ECTMVEKKVLALL or 25 μg of peptide SVEWWAYGLLYEMLA were added to each well using different conditions for each peptide, and PKC activity was measured after 30 minutes of incubation. Saline vehicle was used as negative control, and 10 mM insulin was used as positive control. [Diagram 2]Figure 1 shows the expression levels of FATP isoforms in human adipocytes treated with peptides derived from human alpha PKC. After 48 hours of incubation with different peptides derived from alpha PKC (i.e., ECTMVEKKVLALL and SVEWWAYGLLYEMLA), the expression levels of the six isoforms of FATP were measured by real-time PCR. Normalized expression levels of the six isoforms of FATP (Fatp1-6) in human adipocytes. GAPDH was used as the reference gene. [Diagram 3] In vitro dose response of PATAS to induce PKC activity in human adipocytes. In vitro dose response of PATAS to induce PKC activity in human adipocytes after 30 min incubation of primary human adipocytes with PATAS in culture medium. n=5 / group, amount of PATAS is expressed in μg per well. [Figure 4] Glucose uptake in human primary mature adipocytes. N=8 / group. ADPIF peptide induces glucose uptake in human primary mature adipocytes with efficacy comparable to that of insulin. [Figure 5A] Figure 1. ADPIF peptide is more active than PATAD peptide in improving glucose intolerance in mice. Glucose tolerance: Mice were injected with either vehicle (saline), vehicle + PATAD417, or vehicle + ADPIF (CPC peptide A-MRO) on day 0. Six days later (D6), mice were fasted for 4 hours and ipGTT was performed with glucose subcutaneously injected at 0 min. Glucose concentrations were measured from tail blood every 30 min. Control mice were kept on a chow diet (CTL Chow diet) and all other mice were kept on a high fat / high glucose diet. [Figure 5B]Figure 1 shows that ADPIF peptide is more active than PATAD peptide in improving glucose intolerance in mice. The corresponding area under the curve (AUC) of the glucose tolerance test shown in (A) shows that AUC decreased upon injection of PATAD (PATAD417) and ADPIF (PATAD417-MRP), indicating that ADPIF is more effective at decreasing AUC than PATAD. This indicates that ADPIF is more effective at improving glucose intolerance than PATAD. [Figure 6A] Figure 1. PATAS has antidiabetic effects in an established mouse model of the disease (db / db; BKS from Jax lab) along with decreased expression levels of FATP2 in adipose tissue. Blood glucose excursion curves in ipGTT on day 4 after subcutaneous PATAS injection (2mg / kg BW PATAS dose) in 6-week-old db / db male mice on a chow diet after 8 hours of fasting. Glucose bolus (2g / kg body weight) administered subcutaneously at T=0 min and associated AUC histograms (n=10 mice / group, significance set at *p-value ≤ 0.05, **p-value ≤ 0.01). [Figure 6B] Figure 1. PATAS has antidiabetic effects in an established disease mouse model (db / db; BKS from Jax lab) with reduced expression levels of FATP2 in adipose tissue. db / db mice sWAT after 4 weeks of PATAS injections and after an additional 4 weeks of no treatment. Normalized expression levels of the six FATP isoforms (FATP1-6) in subcutaneous white adipose tissue (sWAT) were determined. GAPDH was used as the reference gene, n=6 / group. [Figure 7A]Figure 7A shows that PATAS is effective in improving glucose intolerance in a genetic mouse model of Bardet-Biedl syndrome (BBS), a rare disease associated with obesity and type 2 diabetes. BBS mice were fed a chow diet ad libitum. n=4 mice were used in a crossover experimental setup and ipGTT was performed before and 3 days after injection. BBS10 gene knockout model mice (Bbs10- / -) were generated and allowed to become naturally obese as described in the literature. At 4 months of age, intraperitoneal glucose tolerance test (ipGTT) was performed before PATAS administration in Bbs10- / - mice, which showed that Bbs10- / - mice were glucose intolerant (Figure 7A). The same mice were administered PATAS at 2 mg / kg body weight into the subcutaneous adipose tissue and 3 days later, ipGTT showed that Bbs10- / - mice had improved glucose tolerance (Figure 7A). [Figure 7B] The area under the curve (AUC) corresponding to Figure 7A was ~30000 mg / dL min (Figure 7B). We found that the corresponding AUC decreased to ~16000 mg / dL min (Figure 7B). [Figure 8] Figure 1. Effect of ADPIF on glycated albumin in plasma in the STAM mouse model. Plasma from treated mice in the Japanese STAMT model was used to measure circulating glycated albumin levels using a commercial kit. Results are shown as the average of six mice per group, and error bars indicate the standard error of the mean in this figure. After injecting the corresponding mice with ADPIF once a week for 5 weeks, it was measured that glycated albumin levels were significantly reduced compared to the vehicle-injected mouse group. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0032] Surprisingly, the present inventors have provided a peptide and its derivatives derived from the kinase domain of PKCα that specifically reduces the expression of FATP2 (Fatty acid transport protein 2) in adipose tissue. Finally, the peptide can reduce glycated albumin in plasma, which is a biomarker for diabetes.

[0033] Thus, the peptides described herein are useful in the treatment of diabetes and related disorders.

[0034] Diabetes mellitus is characterized by hyperglycemia. More specifically, type 2 diabetes is characterized by hyperglycemia and insulin resistance. Obesity is believed to be the main cause of type 2 diabetes in people who are genetically predisposed to it. Also, diabetic retinopathy, diabetic neuropathy, and diabetic nephropathy are well-known disorders associated with diabetes and insulin resistance. Increasing glucose uptake and lowering blood glucose can then treat or slow the progression or onset of these diseases.

[0035] Thus, the present invention provides - a peptide as defined herein for use in the treatment of diabetes and related disorders; - a pharmaceutical composition comprising a peptide as defined herein for use in the treatment of diabetes and related disorders; - the use of a peptide or a pharmaceutical composition as defined herein for the manufacture of a medicament for the treatment of diabetes and related disorders; - a method of treating diabetes and related disorders in a subject, comprising administering a therapeutically effective amount of a peptide as defined herein. Regarding.

[0036] definition ALMS1 (Alström syndrome protein 1) is a protein encoded by the ALMS1 gene. Mutations in the ALMS1 gene are known to cause Alström syndrome. It is described in several databases, namely UniProt ID number Q8TCU4, Gene ID number 7840, and HGNG ID number 428. The reference sequences are disclosed in Genbank under NM_015120.4 for mRNA and NP_055935.4 for protein.

[0037] The terms "protein kinase C" and "PKC" (EC 2.7.11.13) are equivalent and refer to a family of protein kinase enzymes involved in regulating the function of other proteins by phosphorylating hydroxyl groups on serine and threonine amino acid residues on those proteins. PKC is commonly activated by signals such as increased concentrations of diacylglycerol (DAG) or calcium ions (Ca2+). PKC plays an important role in several signaling cascades.

[0038] The PKC family contains at least 15 isozymes in humans and is divided into three major subfamilies: conventional (or classical) PKC, novel PKC, and atypical PKC.

[0039] Conventional (c) PKCs include the isoforms α, βI, βII, and γ. Activation of these PKCs requires Ca 2+ Phospholipids such as DAG and phosphatidylserine are required.

[0040] Novel (n)PKCs include the isoforms δ, ε, η, and θ. These PKCs require DAG but do not require Ca2+ for activation.

[0041] Atypical (a) PKCs include the zeta, iota, and lambda isoforms. These PKCs do not require Ca2+ or diacylglycerol for activation.

[0042] Protein kinase C alpha type, also called αPKC, PKC-A or PKC-alpha, belongs to a family of serine and threonine specific protein kinases that are activated by calcium and the second messenger diacylglycerol. It is described in several databases, namely UniProt ID number P17252, Gene ID number 9393, HGNG ID number 5578, etc. Reference sequences are disclosed in Genbank under NM_02737.2 for mRNA and NP_002728.1 for protein. The protein sequence of human αPKC is disclosed in SEQ ID NO:1.

[0043] The kinase domain of αPKC is from position 339 to position 595 as disclosed in SEQ ID NO:1, and is shown in SEQ ID NO:2.

[0044] "Consists of," "consists essentially of," or "substantially comprises": The description herein of any aspect or embodiment of the invention using such terms with reference to one or more elements is intended to provide support for similar aspects or embodiments of the invention, unless otherwise indicated or clearly contradicted by context, to the extent that the particular element or elements "consists of," "consists essentially of," or "substantially comprises." For example, a peptide or protein described herein as comprising a particular sequence should be understood to also describe a peptide or protein consisting of that sequence, unless otherwise indicated or clearly contradicted by context. By "consists essentially of" it is intended that the peptide or protein consists of the sequence, but may contain 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 substitutions, additions, deletions, or combinations thereof, preferably 1, 2, 3, 4, or 5 substitutions, additions, deletions, or combinations thereof. In particular, by "essentially consist in" it may be intended that the peptide may contain 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 additional amino acids at the N-terminus and / or C-terminus, preferably 1, 2, 3, 4, or 5 additional amino acids, and / or 1, 2, or 3 substitutions, deletions, additions, or combinations thereof. Preferably, the number of substitutions, additions, deletions, or combinations thereof depends on the length of the sequence. For example, the percentage of substitutions, deletions, additions, or combinations thereof may be at most 30%, preferably at most 25%.

[0045] As used herein, the term "substitution" refers to the replacement of a single amino acid with another amino acid in a peptide sequence.

[0046] As used herein, the term "deletion" refers to the removal of a single amino acid in a peptide sequence.

[0047] As used herein, the terms "insertion" or "addition" are equivalent and refer to the addition of a single amino acid in a peptide sequence.

[0048] "Substitution, addition, deletion" refers to the substitution, addition, or deletion of one amino acid. Then, when referring to "1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 substitutions, additions, deletions, or a combination thereof," "1, 2, 3, 4, or 5 substitutions, additions, deletions, or a combination thereof," or "1, 2, or 3 substitutions, deletions, additions, or a combination thereof," it means "1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 modifications of amino acids selected from substitutions, additions, deletions, and combinations thereof," "1, 2, 3, 4, or 5 modifications of amino acids selected from substitutions, additions, deletions, or combinations thereof," or "1, 2, or 3 modifications of amino acids selected from substitutions, deletions, additions, or combinations thereof." Also, "1, 2, 3, 4, or 5 substitutions, additions, deletions, or a combination thereof" means "1 to 5 substitutions, additions, deletions, or a combination thereof." Furthermore, "one, two, or three substitutions, additions, deletions, or a combination thereof" means "one to three substitutions, additions, deletions, or a combination thereof."

[0049] In the peptide sequences disclosed herein, amino acids are represented by their one-letter codes according to the following nomenclature: A: alanine, C: cysteine, D: aspartic acid, E: glutamic acid, F: phenylalanine, G: glycine, H: histidine, I: isoleucine, K: lysine, L: leucine, M: methionine, N: asparagine, P: proline, Q: glutamine, R: arginine, S: serine, T: threonine, V: valine, W: tryptophan, Y: tyrosine.

[0050] As used herein, the term "sequence identity" or "identity" refers to the exact correspondence between the amino acids of two peptides. The percentage of identity can be determined by a direct comparison of the sequence information between two molecules by aligning the sequences, counting the exact number of matches between the two aligned sequences, dividing by the length of the shorter sequence, and multiplying the result by 100.

[0051] Sequence identity can be determined by aligning two peptide sequences using a global or local alignment algorithm, depending on the length of the two sequences. Sequences of similar length are preferably aligned using a global alignment algorithm (e.g., Needleman Wunsch) that optimally aligns the sequences over their entire length, while sequences of substantially different lengths are preferably aligned using a local alignment algorithm (e.g., Smith Waterman). Sequences may then be called "substantially identical" or "essentially similar" if they share at least a certain minimum percentage of sequence identity (e.g., when optimally aligned using default parameters in programs such as GAP or BESTFIT). GAP uses the Needleman and Wunsch global alignment algorithm to align two sequences over their entire length (full length), maximizing the number of matches and minimizing the number of gaps. Global alignment is suitable for use in determining sequence identity when the lengths of two sequences are similar.

[0052] "Increased", "increase" or "enhance" is intended to refer to a measurement that is increased by at least 10, 20, 30, 40, 50, 60, 70, 80 or 90% compared to a measurement measured in the absence of the test molecule under the same conditions. "Decreased" or "reduction" is intended to refer to a measurement that is decreased by at least 10, 20, 30, 40, 50, 60, 70, 80 or 90% compared to a measurement measured in the absence of the test molecule under the same conditions.

[0053] As used herein, the terms "treatment," "treat," or "treating" refer to any action intended to improve the health status of a patient, such as curing, mitigating, or delaying disease, and include not only curative but also prophylactic treatment.

[0054] The term treatment specifically designates the correction, delay or reduction of impaired glucose homeostasis. The term "treatment" also designates the improvement of glucose uptake (e.g., glucose sequestration by adipocytes). In the context of the present invention, the term "controlling blood glucose levels" or "control of blood glucose levels" refers to the normalization or regulation of blood or plasma glucose levels in a mammalian subject having abnormal levels (i.e., levels below or above a known baseline, median or mean value of a corresponding mammalian subject having normal glucose homeostasis).

[0055] As used herein, the term "effective amount" refers to an amount of a peptide of the present disclosure or a pharmaceutical composition of the present disclosure that treats or delays the progression or onset of diabetes or a related disorder. It can also refer to an amount of a peptide of the present disclosure or a pharmaceutical composition of the present disclosure that treats or delays diabetes or a related disorder.

[0056] As used herein, the terms "active principle", "active ingredient" and "active pharmaceutical ingredient" are equivalent and refer to the component of a pharmaceutical composition that has a therapeutic effect.

[0057] As used herein, the term "therapeutic effect" refers to an effect induced by an active ingredient, such as a peptide of the present disclosure, or a pharmaceutical composition according to the present disclosure, that can treat or delay the progression or onset of diabetes or related disorders.

[0058] As used herein, the term "excipient or pharma- ceutically acceptable carrier" refers to any ingredient, other than the active ingredient, present in a pharmaceutical composition. Its addition may be intended to impart a particular consistency or other physical or taste characteristics to the final product. An excipient or pharma-ceutically acceptable carrier must be free of interactions, especially chemical interactions, with the active ingredient.

[0059] As used herein, the terms "subject," "individual," or "patient" are used interchangeably to refer to an animal, preferably a mammal, and more preferably a human, including adults, children, newborns, and prenatal humans.

[0060] As used herein, the term "about" refers to values ​​within a range of ±10% of the stated value. For example, "about 50" includes values ​​within ±10% of 50, i.e., values ​​ranging from 45 to 55. Preferably, the term "about" refers to values ​​within a range of ±5% of the stated value.

[0061] peptide The peptides according to the present disclosure have the following characteristics: - does not contain one methionine, one proline and one arginine at the same time; - preferably adopts a secondary structure which is a helix, preferably an alpha helix; - comprising, consisting essentially of, or consisting of a sequence derived from a segment of the kinase domain of PKC (protein kinase C), preferably a segment of at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 25 consecutive residues of the kinase domain of PKC (protein kinase C), or a segment of 5 to 40 consecutive residues of the kinase domain of PKC (protein kinase C); and - the peptide sequence may contain 1, 2, 3, 4 or 5 amino acid modifications selected from substitutions, deletions, additions and combinations thereof within said sequence of a segment of the kinase domain of PKC; Shows.

[0062] The peptides further have the following characteristics: - having a length of less than 80 amino acids, more preferably less than 60 amino acids, even more preferably less than 40 amino acids, even more preferably less than 30 amino acids; - having a length of at least 5 amino acids and less than 40 amino acids, preferably having a length of at least 5, 6, 7, 8 or 9 amino acids and less than 30 amino acids, more preferably having a length of at least 5, 6, 7, 8 or 9 amino acids and less than 25 amino acids; - modified by cross-linking; - being able to inhibit the ALMS1-PKC interaction, in particular being able to reduce or prevent the interaction between ALMS1 and αPKC; or being unable to inhibit the ALMS1-PKC interaction, in particular being able to reduce or prevent the interaction between ALMS1 and αPKC; - Altering the expression levels of FATP in adipose tissue and preferentially decreasing the expression of FATP2 in adipose tissue may represent one or more of:

[0063] The peptides further have the following characteristics: - having a length of less than 80 amino acids, more preferably less than 60 amino acids, even more preferably less than 40 amino acids, even more preferably less than 30 amino acids; - have a length of at least 5, 6, 7, 8 or 9 amino acids and less than 40 amino acids, preferably have a length of at least 5, 6, 7, 8 or 9 amino acids and less than 30 amino acids, more preferably have a length of at least 5, 6, 7, 8 or 9 amino acids and less than 25 amino acids; - modified by cross-linking; - inability to inhibit the ALMS1-PKC interaction, in particular to reduce or prevent the interaction between ALMS1 and αPKC may represent one or more of:

[0064] In one aspect, the peptide of the present disclosure comprises, consists essentially of, or consists of a sequence derived from a segment of the kinase domain of PKC (protein kinase C). The PKC can be selected from conventional PKC, novel PKC, and atypical PKC. In particular, the PKC can be selected from conventional PKC. Preferably, the PKC can be selected from the group consisting of α, βI, βII, and γPKC. More preferably, the PKC can be selected from the group consisting of α, βI, and βIIPKC. Even more preferably, the PKC is αPKC, preferably human αPKC, more preferably human αPKC of SEQ ID NO: 1. The kinase domain of human αPKC is disclosed in SEQ ID NO: 2.

[0065] A segment of a kinase domain of PKC comprises at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 25 consecutive residues of a kinase domain of PKC. In one embodiment, a segment of a kinase domain of PKC comprises 5-40 consecutive residues (optionally 5-30, or 5-25, or 7-25, or 8-25, or 9-25, or 10 to 25, or 11-25, or 12-25) of a kinase domain of PKC.

[0066] The kinase domain of the PKC from which the segment is selected preferably has at least 40% identity with the sequence of SEQ ID NO:2, more preferably at least 45, 50, 55, 60, 65, 70, 75, 80, 85, 90 or 95% identity with the sequence of SEQ ID NO:2.

[0067] Preferably, said sequence of a segment of the kinase domain of PKC represents at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99% of the peptide sequence. In a particular embodiment, the peptide sequence according to the present disclosure consists of the sequence of a segment of SEQ ID NO:1.

[0068] If the segment of the kinase domain of PKC contains one methionine and / or one proline and / or one arginine, then the sequence can be modified (i.e., by introducing substitutions) to remove all proline residues and / or all methionine residues and / or all arginine residues. For example, the sequence can be modified (i.e., by introducing substitutions) to remove all proline residues. Alternatively, the sequence can be modified (i.e., by introducing substitutions) to remove all methionine residues. Otherwise, the sequence can be modified (i.e., by introducing substitutions) to remove all arginine residues. In one embodiment, the sequence can be modified (i.e., by introducing substitutions) to remove all proline and methionine residues. In another embodiment, the sequence can be modified (i.e., by introducing substitutions) to remove all proline and arginine residues. In an additional embodiment, the sequence can be modified (i.e., by introducing substitutions) to remove all methionine and arginine residues. More preferably, the sequence can be modified (ie, by introducing substitutions) to remove all proline residues, all methionine residues, and all arginine residues.

[0069] Preferably, the peptide comprises no more than 20, preferably no more than 15, more preferably no more than 10 amino acid modifications selected from substitutions, deletions, additions, and combinations thereof. In particularly preferred embodiments, the peptide may comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid modifications selected from substitutions, deletions, additions, and combinations thereof, preferably 1, 2, 3, 4, or 5, more preferably 1, 2, or 3.

[0070] For example, the peptide has at least 70%, 75%, 80%, 85%, 90%, 95%, 99% identity with the sequence of a segment of the kinase domain PKC, preferably with the sequence of SEQ ID NO: 2. In one embodiment, a portion of the sequence of the peptide corresponding to SEQ ID NO: 2 has at least 70%, 75%, 80%, 85%, 90%, 95% identity with the sequence of the segment of SEQ ID NO: 2.

[0071] In a particular embodiment, the peptide has at least one amino acid substituted, deleted or added compared to the sequence of a segment of the kinase domain PKC, preferably the sequence of SEQ ID NO:2.

[0072] For example, the sequence of a segment of the kinase domain of PKC may belong to the sequence between positions 339 and 432 of SEQ ID NO:1, between positions 434 and 544 of SEQ ID NO:1, between positions 546 and 561 of SEQ ID NO:1, between positions 563 and 565 of SEQ ID NO:1, or between positions 568 and 595 of SEQ ID NO:1.

[0073] In one embodiment, the sequence of a segment of the kinase domain of PKC may exclude the following residues: G433, E545, S562, S566 of SEQ ID NO:1.

[0074] In one embodiment, the peptides of the present disclosure have an alpha-helical structure. As used herein, the terms "alpha-helix", "α-helix", "classical Pauling-Corey-Branson α-helix" and "3.613-helix" are equivalent and refer to each other. The term "alpha-helix" refers to a common motif in protein secondary structure, a right-handed or helical structure (helix) in which all backbone NH groups provide hydrogen bonds to the C=O group of the amino acid backbone located 3 or 4 residues earlier in the protein sequence. An alpha-helix has an average of about 3.6 residues per helix turn, and 13 atoms are involved in the ring formed by hydrogen bonds.

[0075] In certain embodiments, the peptides of the present disclosure have an alpha-helical structure and / or have a sequence that predicts an alpha-helical structure. Methods for determining the structure of a peptide are well known to those of skill in the art, such as circular dichroism and NMR. Similarly, methods for predicting the alpha-helical structure of a peptide are well known to those of skill in the art, such as STRIDE (Frishman D., Argos P., Proteins, Vol. 23, No. 4, 1995, pp. 566-579), DEFINE (Richards FM, Kundrot CE, Proteins, Vol. 3, No. 2, 1988, pp. 71-84), DSSP (Touw et al., Nucleic Acids Research 2015, 43:D364-D368; Kabsch and Sander., Biopolymers., 1983, 22, 2577-2637), and the like.

[0076] The alpha helices are at the following positions in the kinase domain: 372-377, 381-392, 425-432, 437-456, 466-468, 502-504, 507-510, 518-533, 543-552, 563-572, 577-579, 587-593 and 595-597 of SEQ ID NO:1.

[0077] Thus, the peptide has the following sequence: - VECTMVEKRVLA (SEQ ID NO: 3); - LMYHIQQV (SEQ ID NO: 4); - LDN; - PDY; - PEII (SEQ ID NO:5); - SVDWWAYGVLLYEMLA (SEQ ID NO: 6); - EDEDELFQSIME(sequence number 7); - PAK; - GERDVRE (SEQ ID NO: 8); - AFF It may comprise, consist essentially of, or consist of at least one of:

[0078] In certain embodiments, the peptide has the following sequence: - VECTMVEKRVLA (SEQ ID NO: 3), and - GERDVRE (SEQ ID NO: 8) It may comprise, consist essentially of, or consist of at least one of:

[0079] Optionally, the peptides have the following sequences: VECTMVEKRVLA (SEQ ID NO:3); VECTXVEKRVLA (SEQ ID NO:9); VECTMVEKXVLA (SEQ ID NO:10); VECTXVEKXVLA (SEQ ID NO:11); LMYHIQQV (SEQ ID NO:4); LXYHIQQV (SEQ ID NO:12); LDN; SVDWWAYGVLLYEMLA (SEQ ID NO:6); SVDWWAYGVLLYEXLA (SEQ ID NO:13); SVXWWAYGLLYEMLA (SEQ ID NO:52); EDE The peptide may comprise, consist essentially of, or consist of at least one of: DELFQSIME (SEQ ID NO: 7); EDEDELFQSIXE (SEQ ID NO: 14); GERDVRE (SEQ ID NO: 8); GEXDVRE (SEQ ID NO: 15); GERDVXE (SEQ ID NO: 16); GEXDVXE (SEQ ID NO: 17); LDN; AFF; PDY; XDY; PEII (SEQ ID NO: 5); XEII (SEQ ID NO: 18); PAK; XAK, where X is any amino acid except M, P, and R. Preferably, X is an amino acid that favors an alpha-helical secondary structure. For example, X may be selected from the group consisting of A, D, N, C, G, Q, E, H, L, K, F, S, W, and Y, more preferably A, D, N, G, Q, E, H, L, K, F, S, W, and Y.

[0080] In one aspect, the peptide may comprise, consist essentially of, or consist of at least one of the following sequences: VECTMVEKRVLA (SEQ ID NO:3); VECTXVEKRVLA (SEQ ID NO:9); VECTMVEKXVLA (SEQ ID NO:10); VECTXVEKXVLA (SEQ ID NO:11); LMYHIQQV (SEQ ID NO:4); LXYHIQQV (SEQ ID NO:12); SVDWWAYGVLLYEMLA (SEQ ID NO:6); SVDWWAYGVLLYEXLA (SEQ ID NO:13); SVXWWAYGLLYEMLA (SEQ ID NO:52); EDEDELFQSIME (SEQ ID NO:7); EDEDELFQSIXE (SEQ ID NO:14); GERDVRE (SEQ ID NO:8); GEXDVRE (SEQ ID NO:15); GERDVXE (SEQ ID NO:16); GEXDVXE (SEQ ID NO:17), where X is any amino acid except M, P, and R. In certain embodiments, the peptide may comprise, consist essentially of, or consist of VECTMVEKXVLA (SEQ ID NO: 10), where X is K. In certain embodiments, the peptide may comprise, consist essentially of, or consist of SVXWWAYGLLYEMLA (SEQ ID NO: 52), where X is E.

[0081] In particular, the peptide may comprise, consist essentially of, or consist of at least one of the following sequences: VECTMVEKRVLA (SEQ ID NO:3); VECTXVEKRVLA (SEQ ID NO:9); VECTMVEKXVLA (SEQ ID NO:10); VECTXVEKXVLA (SEQ ID NO:11); LXYHIQQV (SEQ ID NO:12); SVDWWAYGVLLYEXLA (SEQ ID NO:13); EDEDELFQSIXE (SEQ ID NO:14); GERDVRE (SEQ ID NO:8); GEXDVRE (SEQ ID NO:15); GERDVXE (SEQ ID NO:16); GEXDVXE (SEQ ID NO:17), where X is any amino acid except M, P, and R. For example, the peptide may comprise at least one of the following sequences: VECTMVEKRVLA or VECTTVEKEVLA (SEQ ID NO:19).

[0082] Optionally, the peptide comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 substitutions, deletions, additions, or combinations thereof, preferably 1, 2, 3, 4, or 5 substitutions, deletions, additions, or combinations thereof, more preferably 1, 2, or 3 substitutions.

[0083] Optionally, the peptide has the following sequence: VECTMVEKRVLA (SEQ ID NO:3), optionally with 1, 2, 3, 4, or 5 modifications of amino acids selected from substitution, deletion, addition, and combinations thereof, more preferably with 1, 2, or 3 modifications of amino acids selected from substitution, deletion, addition, and combinations thereof; VECTXVEKRVLA (SEQ ID NO:9), optionally with 1, 2, 3, 4, or 5 amino acid modifications selected from substitution, deletion, addition, and combinations thereof, more preferably with 1, 2, or 3 amino acid modifications selected from substitution, deletion, addition, and combinations thereof; VECTMVEKXVLA (SEQ ID NO:10), optionally with 1, 2, 3, 4, or 5 amino acid modifications selected from substitution, deletion, addition, and combinations thereof, more preferably with 1, 2, 3, 4, or 5 amino acid modifications selected from substitution, deletion, addition, and combinations thereof. VECTXVEKXVLA (SEQ ID NO:11), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof; LMYHIQQV (SEQ ID NO:4), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof; LXYHIQQV (SEQ ID NO:12), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof; LDN; optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof. SVDWWAYGVLLYEMLA (SEQ ID NO: 6), optionally having 1, 2, 3, 4, or 5 amino acid modifications selected from substitution, deletion, addition, and combinations thereof, more preferably having 1, 2, or 3 amino acid modifications selected from substitution, deletion, addition, and combinations thereof; SVDWWAYGVLLYEXLA (SEQ ID NO: 13), optionally having 1, 2, 3, 4, or 5 amino acid modifications selected from substitution, deletion, addition, and combinations thereof, more preferably having 1, 2, or 3 amino acid modifications selected from substitution, deletion, addition, and combinations thereof;SVXWWAYGLLYEMLA (SEQ ID NO:52), optionally with 1, 2, 3, 4, or 5 amino acid modifications selected from substitution, deletion, addition, and combinations thereof, more preferably with 1, 2, or 3 amino acid modifications selected from substitution, deletion, addition, and combinations thereof; EDEDELFQSIME (SEQ ID NO:7), optionally with 1, 2, 3, 4, or 5 amino acid modifications selected from substitution, deletion, addition, and combinations thereof, more preferably with 1, 2, or 3 amino acid modifications selected from substitution, deletion, addition, and combinations thereof; EDEDELFQSIXE (SEQ ID NO:8), optionally with 1, 2, 3, 4, or 5 amino acid modifications selected from substitution, deletion, addition, and combinations thereof, more preferably with 1, 2, or 3 amino acid modifications selected from substitution, deletion, addition, and combinations thereof; SEQ ID NO:14); GERDVRE (SEQ ID NO:8), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof; GEXDVRE (SEQ ID NO:15), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof; GERDVXE (SEQ ID NO:16), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof; GEXDVXE (SEQ ID NO:17), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof; LDN; AFF; PDY; XDY; PEII (SEQ ID NO:5); XEII (SEQ ID NO:18); PAK; XAK, where X is any amino acid except M, P and R. Preferably, X is an amino acid that favors an alpha-helical secondary structure. For example, X may be selected from the group consisting of A, D, N, C, G, Q, E, H, L, K, F, S, W and Y, more preferably A, D, N, G, Q, E, H, L, K, F, S, W and Y;

[0084] In one aspect, the peptide has the following sequence: VECTMVEKRVLA (SEQ ID NO:3), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition, and combinations thereof; VECTXVEKRVLA (SEQ ID NO:9), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition, and combinations thereof; VECTMVEKXVLA (SEQ ID NO:10), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition, and combinations thereof. row number 10); VECTXVEKXVLA (SEQ ID NO:11), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof; LMYHIQQV (SEQ ID NO:4), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof; LXYHIQQV (SEQ ID NO:12), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof; optionally with substitution, deletion, addition and combinations thereof. SVDWWAYGVLLYEMLA (SEQ ID NO: 6), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof; SVDWWAYGVLLYEXLA (SEQ ID NO: 13), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof; SVXWWAYGLLYEMLA (SEQ ID NO: 52), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof; EDEDELFQSIME (SEQ ID NO: 7), optionally with 1, 2, or 3 amino acid modifications selected from substitution, deletion, addition, and combinations thereof; EDEDELFQSIXE (SEQ ID NO: 14), optionally with 1, 2, or 3 amino acid modifications selected from substitution, deletion, addition, and combinations thereof; GERDVRE (SEQ ID NO: 8), optionally with 1, 2, or 3 amino acid modifications selected from substitution, deletion, addition, and combinations thereof; GEXDVRE (SEQ ID NO: 15), optionally with 1, 2, or 3 amino acid modifications selected from substitution, deletion, addition, and combinations thereof;It may comprise, consist essentially of, or consist of at least one of: GERDVXE (SEQ ID NO: 16), optionally with one, two, or three amino acid modifications selected from substitution, deletion, addition, and combinations thereof; GEXDVXE (SEQ ID NO: 17), optionally with one, two, or three amino acid modifications selected from substitution, deletion, addition, and combinations thereof, where X is any amino acid except M, P, and R. Preferably, X is an amino acid that favors an alpha-helical secondary structure. For example, X may be selected from the group consisting of A, D, N, C, G, Q, E, H, L, K, F, S, W, and Y, more preferably, A, D, N, G, Q, E, H, L, K, F, S, W, and Y;

[0085] In particular, the peptides have the following sequences: VECTMVEKRVLA (SEQ ID NO:3), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof; VECTXVEKRVLA (SEQ ID NO:9), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof; VECTMVEKXVLA (SEQ ID NO:10), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof; VECTXVEKXVLA (SEQ ID NO:11), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition and combinations thereof; The polypeptide may comprise, consist essentially of, or consist of at least one of: GERDVRE (SEQ ID NO:8), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition, and combinations thereof; GEXDVRE (SEQ ID NO:15), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition, and combinations thereof; GERDVXE (SEQ ID NO:16), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition, and combinations thereof; GEXDVXE (SEQ ID NO:17), optionally with one, two or three amino acid modifications selected from substitution, deletion, addition, and combinations thereof, where X is any amino acid except M, P, and R. Preferably, X is an amino acid that favors an alpha-helical secondary structure. For example, X may be selected from the group consisting of A, D, N, C, G, Q, E, H, L, K, F, S, W and Y, more preferably A, D, N, G, Q, E, H, L, K, F, S, W and Y.

[0086] For example, the peptide may include at least one of the following sequences: VECTMVEKRVLA or VECTTVEKEVLA (SEQ ID NO: 19).

[0087] In one embodiment, the peptide has the following sequence: a) VECTXVEKXVLALLDKXXFLTQLHS (SEQ ID NO: 20), optionally with 1, 2, 3, 4 or 5 amino acid modifications selected from substitution, deletion, addition and combinations thereof, more preferably with 1, 2 or 3 amino acid modifications selected from substitution, deletion, addition and combinations thereof, where X is any amino acid except M, P and R, preferably an amino acid that favors an alpha helical secondary structure, more preferably an amino acid selected from the group consisting of A, D, N, C, G, Q, E, H, L, K, F, S, W and Y, even more preferably A, D, N, G, Q, E, H, L, K, F, S, W and Y; b) VECTMVEKRVLALLDKXXFLTQLHS (SEQ ID NO: 21), optionally with 1, 2, 3, 4 or 5 amino acid modifications selected from substitution, deletion, addition and combinations thereof, more preferably with 1, 2 or 3 amino acid modifications selected from substitution, deletion, addition and combinations thereof, where X is any amino acid except M, P and R, preferably an amino acid that favors an alpha helical secondary structure, more preferably an amino acid selected from the group consisting of A, D, N, C, G, Q, E, H, L, K, F, S, W and Y, even more preferably A, D, N, G, Q, E, H, L, K, F, S, W and Y; c) VECTXVEKRVLALLDKPPFLTQLHS (SEQ ID NO: 22), optionally with 1, 2, 3, 4, or 5 amino acid modifications selected from substitution, deletion, addition, and combinations thereof, more preferably with 1, 2, or 3 amino acid modifications selected from substitution, deletion, addition, and combinations thereof, where X is any amino acid except M, P, and R, preferably an amino acid that favors an alpha-helical secondary structure, more preferably an amino acid selected from the group consisting of A, D, N, C, G, Q, E, H, L, K, F, S, W, and Y, even more preferably an amino acid selected from the group consisting of A, D, N, G, Q, E, H, L, K, F, S, W, and Y; d) VECTMVEKXVLALLDKPPFLTQLHS (SEQ ID NO: 23), optionally with 1, 2, 3, 4 or 5 amino acid modifications selected from substitution, deletion, addition and combinations thereof, more preferably with 1, 2 or 3 amino acid modifications selected from substitution, deletion, addition and combinations thereof, where X is any amino acid except M, P and R, preferably an amino acid that favors an alpha helical secondary structure, more preferably an amino acid selected from the group consisting of A, D, N, C, G, Q, E, H, L, K, F, S, W and Y, even more preferably A, D, N, G, Q, E, H, L, K, F, S, W and Y; and a sequence of any segment of at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 25 consecutive residues from any of the sequences a) to d). It may comprise, consist essentially of, or consist of at least one of:

[0088] In another particular embodiment, a peptide according to the present disclosure is designed or modified to maintain it in an alpha-helical conformation. As known in the art, this can be achieved through a variety of methods, including modifying the amino acid sequence with substitutions of amino acids that are not important for the biological effect, using non-natural amino acids, cyclizing the peptide, and modifying the peptide backbone or adding chemical bonds between amino acids in the peptide chain. Such modifications can be made, for example, to increase the thermostability or protease stability of the peptide.

[0089] In particular, the peptides of the present disclosure are modified by chemical cross-linking. For example, the peptides can be stapled peptides. In one embodiment, the peptides of the present disclosure are stapled. As used herein, the term "stapled peptide" or "stitched peptide" refers to an artificially modified peptide in which the secondary structure of the peptide is stabilized with one or more artificial molecular bridges that connect adjacent turns of the α-helix of the peptide. Methods for preparing stapled peptides are well known in the art and are described, for example, in Verdine and Hilinski (2012, Methods Enzymol, 503, 3-33), WO10033617 and WO10011313, the disclosures of which are incorporated herein by reference.

[0090] In one embodiment, the crosslinks of the stapled peptides of the present disclosure are i+3 and / or i+4 and / or i+7 crosslinks. In a peptide, an "i+3 crosslink" is a crosslink between an "i" amino acid and another amino acid that is 3 amino acid residues away from the i amino acid. In a peptide, an "i+4 crosslink" is a crosslink between an "i" amino acid and another amino acid that is 4 amino acid residues away from the i amino acid. In a peptide, an "i+7 crosslink" is a crosslink between an "i" amino acid and another amino acid that is 7 amino acid residues away from the i amino acid. In a preferred embodiment, the peptide has an "i+7 crosslink".

[0091] For the shortest sequences, especially those containing 3-4 residues, bridges are i+3 and i+4, introduced between residues outside this sequence. If the sequence is long enough, bridge i+7 is preferred.

[0092] To illustrate this embodiment with one particular peptide, the peptide has the following sequence:

[0093] [ka]

[0094] may comprise, consist essentially of, or consist of one of In the formula, the bold and underlined

[0095] [ka]

[0096] the residues of which comprise the staple and are any amino acid derivative suitable for stapling; X is any amino acid except M, P, and R, preferably an amino acid that favors an alpha helical secondary structure, more preferably selected from the group consisting of A, D, N, C, G, Q, E, H, L, K, F, S, W, and Y, even more preferably A, D, N, G, Q, E, H, L, K, F, S, W, and Y; The sequence optionally has 1, 2, 3, 4, or 5 amino acid modifications selected from substitution, deletion, addition, and combinations thereof, and more preferably 1, 2, or 3 amino acid modifications selected from substitution, deletion, addition, and combinations thereof.

[0097] For example, for staple i+7, the first

[0098] [ka]

[0099] is a 2-(7-octenyl)amino acid (e.g., 2-(7-octenyl)alanine or 2-(7-octenyl)arginine), and the second

[0100] [ka]

[0101] is a 2-(4-pentenyl)amino acid (e.g., 2-(4-pentenyl)alanine or 2-(4-pentenyl)serine). Specific combinations can include 2-(7-octenyl)alanine and 2-(4-pentenyl)alanine; 2-(7-octenyl)alanine and 2-(4-pentenyl)serine; 2-(7-octenyl)arginine and 2-(4-pentenyl)alanine; or 2-(7-octenyl)arginine and 2-(4-pentenyl)serine.

[0102] In certain embodiments, the peptide is

[0103] [ka]

[0104] wherein

[0105] [ka]

[0106] have staples, preferably 2-(7-octenyl)arginine and 2-(4-pentenyl)serine, respectively; The sequence optionally has 1, 2, 3, 4, or 5 amino acid modifications selected from substitution, deletion, addition, and combinations thereof, more preferably 1, 2, or 3 amino acid modifications selected from substitution, deletion, addition, and combinations thereof.

[0107] In certain embodiments, the peptide according to the present disclosure is a cyclic peptide. As used herein, the terms "cyclic peptide" or "circular peptide" are equivalent and refer to a peptide in which the N-terminus and C-terminus, or the N-terminus and a side chain of another amino acid, preferably the C-terminal amino acid, or the C-terminus and a side chain of another amino acid, preferably the N-terminal amino acid, or the side chain of an amino acid and a side chain of another amino acid, preferably the N-terminal amino acid and the C-terminal amino acid, are linked by a covalent bond that creates a ring structure. As used herein, the terms "N-terminus", "amino-terminus", "NH2-terminus", "N-terminal end" and "amine-terminus" are equivalent and refer to the free amine group (-NH2) present on the first amino acid of the peptide. As used herein, the terms "C-terminus," "carboxyl-terminus," "carboxy-terminus," "C-terminal end," and "COOH-terminus" are equivalent and refer to the free carboxyl group (-COOH) present on the last amino acid of a peptide.

[0108] In one embodiment, the peptide according to the disclosure has a length of less than 80 amino acids, more preferably less than 60 amino acids, even more preferably less than 40 amino acids, and even more preferably less than 30 amino acids. In a particular embodiment, the peptide according to the disclosure has a length of less than 25 amino acids. In another particular embodiment, the peptide according to the disclosure has a length of less than 20 amino acids, preferably less than 15 amino acids. Preferably, the peptide has a minimum length of more than 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acids. Preferably, the peptide has a length of at least 4 amino acids and less than 40 amino acids, preferably at least 4 amino acids and less than 30 amino acids, and more preferably at least 6 amino acids and less than 25 amino acids.

[0109] In one embodiment, the peptide according to the present disclosure can inhibit the interaction between ALMS1 and PKC, and in particular can reduce or prevent the interaction between ALMS1 and αPKC. That is, the peptide according to the present disclosure can block the interaction between ALMS1 and αPKC. Alternatively, the peptide according to the present disclosure cannot inhibit the interaction between ALMS1 and PKC, and in particular can reduce or prevent the interaction between ALMS1 and αPKC. That is, the peptide according to the present disclosure cannot block the interaction between ALMS1 and αPKC.

[0110] To determine the effect of peptides on the binding of αPKC to ALMS1, any technique known to those skilled in the art can be carried out, in particular any method suitable for determining protein interactions. For example, recombinant or purified native ALMS1 or αPKC can be bound to a surface plasmon resonance chip, and the other molecule can be flowed over the chip to assess binding affinity, for example with a Biacore (General Electric, USA) machine.

[0111] The effect of a peptide on the binding of αPKC to ALMS1 is determined by measuring the binding of αPKC to ALMS1 in the absence and presence of the tested peptide and comparing the binding of αPKC to ALMS1.

[0112] In particular, immunoprecipitation assays can be carried out using ALMS1 as bait. The assays can be carried out using cells, particularly adipocytes, cultured in the absence and / or presence of insulin, preferably in the absence of insulin. The peptide to be tested is added to the culture medium. Then, αPKC is immunodetected.

[0113] "Reduced," "reduction," or "prevention" is intended to refer to binding that is reduced by at least 10, 20, 30, 40, 50, 60, 70, 80, or 90% compared to binding measured in the absence of the test molecule under the same conditions.

[0114] In one embodiment, the peptides according to the present disclosure are capable of reducing the expression of FATP2 in adipose tissue.

[0115] FATP2 is also called Solute Carrier Family 27 Member 2 (SLC27A2). The protein is disclosed in the database UniProtKB under O14975. The gene is listed in the UniGene database under Hs.11729. The reference sequences are listed in NCBI under NP_003636.2 and NM_003645.3 for isoform 1, and NP_001153101.1 and NM_001159629.1 for isoform 2.

[0116] "Decreased" or "reduction" is intended to refer to expression that is decreased by at least 10, 20, 30, 40, 50, 60, 70, 80, or 90% compared to expression measured in the absence of the peptide under the same conditions. Expression can be measured at the protein level (e.g., using an antibody) or at the mRNA level.

[0117] Expression can be measured at the protein level by any available method, such as immunohistochemistry, semi-quantitative Western blot, protein or antibody arrays, etc. Antibodies against FATP2 are commercially available, for example, from Origene (TA350424 or TA333990), Santa Cruz Biotechnology (sc-393906), etc.

[0118] Expression can also be measured at the mRNA level by any available method. Preferably, the expression level of FATP2 is determined by measuring the amount of mRNA transcripts by quantitative RT-PCR, real-time quantitative RT-PCR, Nanostring technology PCR, or by high-throughput sequencing techniques such as RNA-Seq or sequencing techniques using microfluidic systems. More specifically, expression is measured by the methods defined in the Examples section.

[0119] In a particular embodiment, the effect of the peptide on the expression of FATP2 in adipose tissue is preferably specific for FATP2. In this embodiment, the peptide is unable or barely able to affect the expression of other FATPs, namely FATP1, FATP3, FATP4, FATP5 and FATP6, especially in mammalian adipose tissue.

[0120] In certain embodiments, the peptides according to the present disclosure have the following characteristics: - does not contain one methionine, one proline and one arginine at the same time; - adopts a secondary structure which is a helix, preferably an alpha helix; and - comprising, consisting essentially of, or consisting of a sequence derived from a segment of the kinase domain of PKC (protein kinase C); indicates, In addition, it has the following features: - altering the expression levels of FATP in adipose tissue and preferentially decreasing the expression of FATP2 in adipose tissue; - having a length of at least 4 amino acids and less than 40 amino acids, preferably having a length of at least 4 amino acids and less than 30 amino acids, more preferably having a length of at least 4 amino acids and less than 25 amino acids; - adopting a secondary structure which is a helix, preferably an alpha helix; - Modified by cross-linking Indicates one, two, three, four or all of the following:

[0121] In more particular embodiments, the peptides according to the present disclosure have the following characteristics: - does not contain one methionine, one proline and one arginine at the same time; - having a length of at least 4 amino acids and less than 40 amino acids, preferably having a length of at least 4 amino acids and less than 30 amino acids, more preferably having a length of at least 4 amino acids and less than 25 amino acids; - adopt a secondary structure which is a helix, preferably an alpha helix; Shows.

[0122] In another more particular embodiment, the peptide according to the present disclosure has the following characteristics: - Reducing the expression of FATP2 in adipose tissue; - does not contain one methionine, one proline and one arginine at the same time; - having a length of at least 4 amino acids and less than 40 amino acids, preferably having a length of at least 4 amino acids and less than 30 amino acids, more preferably having a length of at least 4 amino acids and less than 25 amino acids; - adopt a secondary structure which is a helix, preferably an alpha helix; Shows.

[0123] The peptide according to the present disclosure may further comprise a moiety that facilitates its uptake or entry into a cell, in particular a PTD (protein transduction domain). A PTD generally comprises a specific amino acid sequence of 10-20 amino acids (Matsushita and Matsui, (2005), J Mol Med 83, 324-328; Vives et al., Biochimic et Biophysica Acta, 2008, 1786, 126-138). A PTD is mainly composed of basic amino acids such as arginine or lysine, and representative examples of PTDs include arginine-rich peptides such as polyR8 (RRRRRRRR (SEQ ID NO: 33)) or (RRPRRPRRPRRPRRP (SEQ ID NO: 34)), antennapedia or penetratin peptides such as (RQIKIWFQNRRMKWKK (SEQ ID NO: 35)) or HIV-Tat (YGRKKRRQRRR (SEQ ID NO: 36)).

[0124] Peptides according to the present disclosure may be composed of natural and / or unnatural amino acids. The term "unnatural amino acid" is defined as an analog or derivative of a natural amino acid (i.e., alanine, valine, glycine, leucine, isoleucine, lysine, arginine, glutamic acid, glutamine, aspartic acid, asparagine, histidine, tyrosine, phenylalanine, tryptophan, serine, proline, threonine, cysteine, methionine), which has been modified, such as by shortening or lengthening the side chain or by carrying a different functional group. In particular, isomers D and L are contemplated, since isomer D is not reactive to proteases. Furthermore, modifications at some or all of the peptide bonds, particularly with (-CO-NH-), (-CH-NH-), (-NH-CO-), (-CH-O-), (-CH-S-), (-CH-CH-), (-CO-CH-), (-CHOH-CH-), (-N=N-), and / or (-CH=CH-), are contemplated to enhance resistance to proteolysis. -) and an amide group (-CONH2). The peptide may also be a D-retro-inverso sequence of a peptide as disclosed herein. The N-terminus may be modified, in particular with an acetyl group.

[0125] Optionally, the peptides can be PEGylated to enhance stability. Optionally, the peptides can be formulated in non-aqueous protic solvent solutions such as propylene glycol and polyethylene glycol. The peptides can also be packaged in polylactide-co-glycolide microsphere depot formulations. Many sustained release delivery systems exist, many of which are suitable for use in the present disclosure. For example, polymer-based sustained release compositions based on degradable polymers such as PLGA, polylactate, or polyglycolate are suitable, as are lipid-based depot compositions, which are described in WO2005 / 117830 and / or WO2006 / 075124, the disclosures of which are incorporated by reference in their entirety. Formulation of active agents into biodegradable polymer depot formulations is well established and well known in the art, and the peptides of the present disclosure can therefore be formulated therewith using known methods. Preferably, the compositions of the present disclosure can release peptides at functional concentrations for at least one month.

[0126] "Peptide" is intended to refer to a peptide as disclosed above, or a combination of different peptides as disclosed above. For example, 2, 3, 4, 5 or 6 different peptides can be used, preferably 2 or 3, more preferably 2.

[0127] combination The peptides according to the present disclosure can be used in combination with one or more additional active agents, such as, for example, antidiabetic agents, lipid-lowering agents, antiobesity agents, antihypertensive agents, antilipidemic agents, anti-inflammatory agents, and agonists of peroxisome proliferator-activated receptors.

[0128] Thus, the present invention provides - peptides or pharmaceutical compositions comprising the peptides disclosed herein for use in the treatment of diabetes and related disorders, particularly in combination with one or more additional agents disclosed herein; - a pharmaceutical composition comprising a peptide disclosed herein and one or more additional active agents for use in the treatment of diabetes and related disorders; - an article of manufacture, combination preparation or kit comprising a peptide according to the present disclosure and one or more additional agents, particularly as disclosed herein, for simultaneous, separate or sequential use in the treatment of diabetes and related disorders; - use of the peptide in combination with one or more additional active agents to manufacture a medicament for the treatment of diabetes and related disorders; - the use of a peptide as disclosed herein, and in particular one or more additional agents as disclosed herein, for the manufacture of a medicament for the treatment of diabetes and related disorders; - a method of treating diabetes and related disorders in a subject, comprising administering a therapeutically effective amount of a peptide disclosed herein and a therapeutically effective amount of one or more additional active agents; - a method for treating diabetes and related disorders in a subject, comprising administering a therapeutically effective amount of a pharmaceutical composition comprising a peptide as disclosed herein and one or more additional agents, particularly as disclosed herein. Regarding.

[0129] In particular, therapeutically effective or sub-therapeutically effective amounts of one or more additional active agents can be used. "Sub-therapeutic" is intended to refer to, for example, an amount that is 90, 80, 70, 60, 50, 40, 30, 20, or 10% of the conventional therapeutic dose (particularly for the same indication and / or the same route and / or frequency of administration).

[0130] Antidiabetic drugs include, for example, insulin, insulin derivatives and mimetics, sulfonylureas (e.g., chlorpropamide, tolazamide, acetohexamide, tolbutamide, glyburide, glimepiride, glipizide); gliflozins such as empagliflozin and dapagliflozin; glyburide and amaryl; liraglutide (NN2211); insulinotropic sulfonylurea receptor ligands such as meglitinides, e.g., nateglinide and repaglinide; thiazolidinediones (e.g., rosiglitazone (AVANDIA), troglitazone (REZULIN), pioglitazone (ACTOS), balaglitazone, rivoglitazone, netoglitazone, troglitazone, englitazone, enhance insulin action (e.g., by insulin sensitization) and thus promote glucose utilization in peripheral tissues, such as ciglitazone, adaglitazone, darglitazone; protein tyrosine phosphatase-IB (PTP-1B) inhibitors, such as PTP-112; cholesteryl ester transfer protein (CETP) inhibitors, such as torcetrapib, GSK3 (glycogen synthase kinase-3) inhibitors, such as SB-517955, SB-4195052, SB-216763, NN-57-05441 and NN-57-05445; RXR ligands, such as GW-0791 and AGN-194204; sodium-dependent glucose cotransporter inhibitors, such as T-1095 and canagliflozin; Glycogen phosphorylase A inhibitors such as R3401; biguanides such as metformin and other drugs that promote glucose utilization, inhibit glucose production in the liver, and reduce glucose excretion in the intestine; alpha-glucosidase inhibitors such as acarbose and migiitoi, and other drugs that delay carbohydrate digestion and the resulting absorption from the intestine and suppress postprandial hyperglycemia; GLP-1 (glucagon-like peptide-1), exendin-4 and other GLP-1 analogs and GLP-1 mimetics; and DPPIV (dipeptidyl peptidase IV) inhibitors such as vildagliptin. Also, it may be an antidiabetic drug described in Expert Opin Investig Drugs 2003, 12(4):623-633, Figures 1-7.Antidiabetic agents may also include molecules that prevent the binding of αPKC to ALMS1, such as those disclosed in WO2015 / 114062, the disclosure of which is incorporated herein by reference.

[0131] Lipid-lowering agents include, for example, 3-hydroxy-3-methyl-glutaryl coenzyme A (HMG-CoA) reductase inhibitors, such as lovastatin, pitavastatin, simvastatin, pravastatin, cerivastatin, mevastatin, velostatin, fluvastatin, dalvastatin, atorvastatin, rosuvastatin, and rivastatin; squalene synthase inhibitors; FXR (farnesoid X receptor) and LXR (liver X receptor) ligands, such as obeticholic acid; bile acid sequestrants, such as cholestyramine and colesevelam; fibrates; nicotinic acid and aspirin; and the transmembrane G protein-coupled receptor (TGR) 5 agonist aramchol.

[0132] Anti-obesity agents include, for example, orlistat, rimonabant, phentermine, topiramate, quinexa, and lorcaserin.

[0133] Examples of antihypertensive agents include loop diuretics such as ethaclinic acid, furosemide, and torsemide; angiotensin-converting enzyme (ACE) inhibitors such as benazepril, captopril, enalapril, fosinopril, lisinopril, moexipril, perinodopril, quinapril, ramipril, and trandolapril; Na-K-ATPase membrane pump inhibitors such as digoxin; neutral endopeptidase (NEP) inhibitors such as sacubitril; ACE / NEP inhibitors such as omapatrilat, sampatrilat, and fasidotril; angiotensin II antagonists such as candesartan, eprosartan, irbesartan, losartan, telmisartan, and valsartan, especially valsartan, NEP inhibitors such as sacubitril and valsartan, and and angiotensin II antagonist combinations (i.e., Entresto); renin inhibitors such as ditekeren, zankiren, terlaskiren, aliskiren, RO-66-1132 and RO-66-1168; beta-adrenergic receptor blockers such as acebutolol, atenolol, betaxolol, bisoprolol, metoprolol, nadolol, propranolol, sotalol, and timolol; inotropes such as digoxin, dobutamine, and milrinone, calcium channel blockers such as amlodipine, bepridil, diltiazem, felodipine, nicardipine, nimodipine, nifedipine, nisoldipine, and verapamil; aldosterone receptor antagonists, and aldosterone synthase inhibitors, and the like.

[0134] Examples of agonists of peroxisome proliferator activator receptors include fenofibrate, pioglitazone, rosiglitazone, tesaglitazar, BMS-298585, L-796449, compounds specifically described in patent application WO2004 / 103995, i.e., the compounds of Examples 1 to 35 or the compounds specifically described in claim 21, or compounds specifically described in patent application WO03 / 043985, i.e., the compounds of Examples 1 to 7 or the compounds specifically described in claim 19, in particular (R)-1-{4-[5-methyl-2-(4-trifluoromethyl-phenyl)-oxazol-4-ylmethoxy]-benzenesulfonyl}-2,3-dihydro-1H-indole-2-carboxyl or a salt thereof.

[0135] Other drugs of interest include, for example, cenicriviroc, simtuzumab, selonsertib, emricasan, and the like.

[0136] In certain embodiments, the one or more additional active agents used in combination with the peptide may be selected from among GLP-1 analogues such as liraglutide, obeticholic acid, gliflozin, simtuzumab (GS6624), cenicriviroc, aramchol, galectin 3 inhibitors such as GR-MD-02, TGR5 agonists and dual FXR / TGR5 agonists such as INT-777 or INT-767, and emricasan.

[0137] The anti-inflammatory agent can be any agent known to those of skill in the art, such as salicylates, nonsteroidal anti-inflammatory drugs (NSAIDs) such as various forms of ibuprofen, various forms of naproxen, steroidal anti-inflammatory drugs such as corticosteroids, anti-inflammatory anti-TNF alpha antibodies, and combinations thereof.

[0138] The form of the pharmaceutical composition, the route of administration, the dosage, the regimen will of course depend on the condition to be treated, the severity of the disease, the age, weight, and sex of the patient, etc.

[0139] Pharmaceutical or therapeutic compositions of the present disclosure may be formulated for topical, oral, parenteral, intranasal, intravenous, intramuscular, subcutaneous or intraocular administration, and the like.

[0140] The peptides used in the pharmaceutical compositions of the present disclosure are present in a therapeutically effective amount.

[0141] Pharmaceutical compositions containing the peptides are formulated according to standard pharmaceutical practice known to those skilled in the art (Lippincott Williams & Wilkins, 2000 and Encyclopedia of Pharmaceutical Technology, edited by J. Swarbrick and JC Boylan, 1988-1999, Marcel Dekker, New York).

[0142] In one aspect, the present invention provides a stable formulation for parenteral injection of a pharmaceutical composition according to the present disclosure comprising a peptide or a salt thereof, the formulation being dried and then reconstituted in a solvent prior to use. The peptide (or each of the peptides, in embodiments where the formulation comprises two or more peptides) is mixed with a non-volatile buffer and dried to a dry peptide powder. Suitable buffers include, but are not limited to, glycine buffer, citrate buffer, phosphate buffer, and mixtures thereof. In one embodiment, the buffer is a glycine buffer. In another embodiment, the buffer is a mixture of citrate buffer and phosphate buffer. In some embodiments, when the formulation comprises two or more peptides, the first and second buffers are the same. In some embodiments, when the formulation comprises two or more peptides, the first and second buffers are different. Alternatively, the pharmaceutical composition according to the present disclosure may be stored in an aqueous state. The solution may contain further additives or excipients, as appropriate, which must be compatible with the active ingredient and, if not removed during the lyophilization step, also compatible with the route of administration. When administered parenterally, the composition may be injected intradermally, subcutaneously, intramuscularly, or intravenously. Preferably, the composition or peptide is or should be injected subcutaneously, especially into fatty tissue.

[0143] Preferably, it may be placed in a mini-osmotic pump or other controlled delivery device implanted in the body. Preferably, it may be mixed with other compounds to form a depot-type sustained release formulation. The preferred route of administration is subcutaneous injection, for example, using a disposable or multi-unit dispensing device similar to an insulin pen. The peptide may also be administered with a device that allows subcutaneous administration without the use of a needle, a non-invasive system.

[0144] Additionally, the peptides can be administered using any available drug delivery system, and in particular, it is contemplated that the recombinant human hyaluronidase enzyme, rHuPH20, can be used to enable and optimize subcutaneous drug delivery for appropriate co-administration therapy.

[0145] With this technology, some biologics and compounds that are administered intravenously may be administered subcutaneously, or just under the skin, potentially providing a better patient experience and increasing efficiency in the healthcare system by reducing administration times, injection pain, and injection-site reactions.

[0146] In one embodiment, the peptides of the present disclosure may be mixed with other compounds to create a depot-type sustained release formulation, which may then be injected subcutaneously to form a sustained release depot.

[0147] For oral administration, the tablets can be formulated into conventional dosage forms suitable for oral administration, such as tablets, capsules, powders, granules, and liquid preparations such as syrups, elixirs, and concentrated drops. Non-toxic solid carriers and diluents can be used, including, for example, pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharin, talcum, cellulose, glucose, sucrose, magnesium, carbonates, and the like. Compressed tablets also require binders, which are agents that impart adhesion to powder materials. For example, starch, gelatin, sugars such as lactose or dextrose, and natural or synthetic gums can be used as binders. Tablets also require disintegrants to facilitate disintegration of the tablets. Disintegrants include starch, clay, cellulose, algin, gum, and cross-linked polymers. Furthermore, lubricants and glidants are also included in tablets to prevent adhesion of tablet materials to the surface during the manufacturing process and to improve the flow properties of powder materials during manufacturing. Colloidal silicon dioxide is the most commonly used lubricant, and compounds such as talc and stearic acid are the most commonly used lubricants.

[0148] For transdermal administration, the composition may be formulated in a form such as an ointment, cream, or gel, and a suitable penetrant or surfactant to promote penetration, such as dimethylsulfoxide, dimethylacetamide, and dimethylformamide, may be used.

[0149] Transmucosal administration may be by nasal sprays, pulmonary inhalation, or rectal or vaginal suppositories. In one embodiment, the present invention may be administered by the pulmonary route, using either a dry powder or liquid formulation administered using a pulmonary drug delivery device according to methods known in the art. The active compound may be incorporated into any of the known suppository bases, using methods known in the art. Examples of such bases include cocoa butter, polyethylene glycol (carbowax), polyethylene sorbitan monostearate, and mixtures of these with other suitable materials to modify the melting point or dissolution rate.

[0150] Pharmaceutical compositions according to the present disclosure may be formulated to release the active agent substantially immediately upon administration, or at any predetermined time or period after administration.

[0151] Pharmaceutical compositions according to the present disclosure can include one or more peptides of the present disclosure in association with pharma- ceutically acceptable excipients and / or carriers, which are selected according to the mode of administration, as described above.

[0152] In certain embodiments, a pharmaceutical composition according to the present disclosure comprises between 0.01 ng and 10 g of a peptide of the present disclosure. In one embodiment, a pharmaceutical composition according to the present disclosure comprises between 0.1 ng and 1 g of a peptide of the present disclosure.

[0153] All documents cited in this application, including scientific articles and abstracts, published patent applications, issued patents, or other documents, are incorporated herein by reference, including all results, tables, figures, and text from these documents.

[0154] The terms "comprising," "having," "consisting in," and "containing" have different meanings but may be substituted for other terms throughout this specification.

[0155] Further aspects and advantages of the present disclosure are illustrated in the following examples, which should be considered as illustrative and not limiting. EXAMPLES

[0156] Example 1 Effect of ADPIF / PATAS peptide on glucose uptake in human primary mature adipocytes Human primary preadipocytes were cultured in 96-well plates and differentiated into human mature adipocytes. After 3 weeks of post-adipogenic culture, adipocytes were incubated in insulin-free medium for 2 hours. Following this 2-hour insulin fast, the medium was changed back to either insulin-free medium (-INS), insulin-containing medium (+INS), or ADPIF-containing medium (PATAS) and incubated for 30 minutes with 2-DG, a glucose analog used to measure glucose uptake in Abcam kits. Glucose uptake was then measured in eight wells per condition according to the manufacturer's protocol, and the average values ​​were calculated and plotted in a histogram, shown in Figure 1, with error bars representing the standard error of the mean.

[0157] Example 2 The ADPIF / PATAS peptide has a stronger antihyperglycemic effect than the PATAD peptide. Mice were injected with a single dose (25 micrograms per mouse) of either scrambled peptide or PATAD or ADPIF / PATAS. Results from a series of glucose tolerance tests (ipGTT) in DIO glucose-intolerant male mice (Figures 5A and 5B).

[0158] Example 3 ADPIF / PATAS peptide increases PKC kinase activity in human adipocytes with a dose-response effect As shown in FIG. 3, it can be observed that PKC activity increases with increasing amount of ADPIF / PATAS, and reaches the same level of PKC activity as insulin at 25 μg of PATAS per well.

[0159] We also tested two other peptides derived from PKC alpha, namely ECTMVEKKVLALL and SVEWWAYGLLYEMLA, for their effect on PKC kinase activity. As shown in Figure 1, both peptides are able to increase PKC kinase activity in human adipocytes.

[0160] Furthermore, we determined two peptides that were effective against the expression of FATP1 to 6, and found that both peptides reduced the expression of FATP2 in adipocytes (Figure 2).

[0161] Example 4 Effects of ADPIF / PATAS peptide in established diabetic mouse model (db / db; BKS male mice) As shown in FIG. 6A, the ADPIF / PATAS peptide can prevent hyperglycemia.

[0162] Furthermore, in this model, we determined the effect of the ADPIF / PATAS peptide on the expression of FATP1-6 in adipocytes. The ADPIF / PATAS peptide can reduce the expression of FATP2 (Figure 6B).

[0163] The activity of PATAS / ADPIF was tested in a diabetic mouse model, db / db, with a BKS genetic background. Six-week-old mice fed a chow diet were subcutaneously injected with saline vehicle or with PATAS at 2 mg / kg body weight, and 4 days later, ipGTT was performed, showing significant improvement of glucose intolerance in response to PATAS / ADPIF treatment (Figure 6A). The mice were then injected with PATAS / ADPIF or vehicle once a week for an additional 3 weeks, and then left untreated for an additional 4 weeks with constant monitoring. db / db mice were then euthanized and the expression levels of fatty acid transport protein isoforms (Fatps) in adipose tissue were measured, showing that the expression levels of Fatp2 were specifically and significantly reduced in PATAS-treated mice compared to control mice (Figure 6B).

[0164] Example 5 Effect of ADPIF / PATAS peptide in Bardet-Biedl syndrome (BBS) model BBS10 gene knockout model mouse (Bbs10 - / - ) were generated and naturally made obese as described in the literature. At 4 months of age, Bbs10 - / -Before administration of PATAS / ADPIF, intraperitoneal glucose tolerance tests (ipGTTs) were performed. - / - The mice were glucose intolerant (Figure 7) with a corresponding area under the curve (AUC) of ~30,000 mg / dL min. The same mice were administered PATAS at 2 mg / kg body weight into the subcutaneous adipose tissue and then subjected to an ipGTT 3 days later. - / - Mice were found to have improved glucose tolerance (Figure 7) with a corresponding decrease in AUC to ∼16000 mg / dL min−1.

[0165] Example 6 ADPIF / PATAS is effective in reducing glycated albumin in the STAMt mouse model after 5 weeks of weekly injections of 25 μg ADPIF / PATAS per mouse Glycated albumin is well known as a robust parameter for determining the onset of hyperglycemic symptoms in vivo. Therefore, plasma from STAMT mice that were injected with 25 μg of ADPIF / PATAS once a week for 5 weeks was used compared to the saline-injected control group. Glycated albumin was significantly reduced in ADPIF / PATAS-treated mice, indicating that ADPIF / PATAS can ameliorate hyperglycemia (Figure 8).

[0166] Materials and Methods Peptide sequencing, synthesis, and solution preparation PATAD stapled peptide sequence: VE CTM-[2-(4-pentenyl)alanine]-EK RVL A-[2-(4-pentenyl)alanine]-L DKP PFL TQL HS (SEQ ID NO: 49) ADPIF / PATAS stapled peptide sequence:

[0167] [ka]

[0168] During the ceremony,

[0169] [ka]

[0170] have staples, preferably 2-(7-octenyl)arginine and 2-(4-pentenyl)serine, respectively.

[0171] Has staples

[0172] [ka]

[0173] have

[0174] [ka]

[0175] Has staples

[0176] [ka]

[0177] have

[0178] [ka]

[0179] Stapled peptides were used at 95% purity.

[0180] All peptides were dissolved and diluted in sterile saline.

[0181] Glucose uptake assay in human mature adipocytes Human white visceral preadipocytes (Cat. No.: C-12732; PromoCell) were purchased. Preadipocytes were seeded according to the manufacturer's protocol and cultured in Preadipocyte Growth Medium (Cat. No.: C-27410; PromoCell) until confluence. The medium was changed to Preadipocyte Differentiation Medium (Cat. No.: C-27436; PromoCell) and cultured for 2 days to induce adipogenic differentiation. After the differentiation phase, the medium was finally changed to Adipocyte Nutrition medium (Cat. No.: C-27438; PromoCell). For insulin-free cultures, insulin-free Adipocyte Basal Medium (Cat. No.: C-2431; PromoCell) was supplemented with 5 g / L deoxyglucose, 8 μg / mL d-biotin, and 400 ng / mL dexamethasone. After 3 weeks of adipogenic differentiation, cultured mature adipocytes were incubated for 2 hours without insulin, after which the medium was replaced with fresh culture medium containing glucose analogue (2-DG) without insulin (-INS), with insulin (+INS), or without insulin + 2.5 μL of test peptide at a concentration of 10 μg / L in a final volume of 200 μL. After 30 minutes of incubation, cells were processed according to the standard protocol of the commercially available glucose uptake assay kit (colorimetric) from Abcam (catalog number: ab136955).

[0182] For PKC kinase activity testing, ready-to-use mature human primary adipocytes from omental adipose tissue were used in 96-well or 6-well plate format purchased from Zenbio (Cat. No.: OA-1096-3 or OA-1006-3). The PKC kinase activity assay kit (Cat. No.: 139437) and procedure were used according to the manufacturer's protocol.

[0183] In vivo mouse study For the obese mouse model that developed type 2 diabetes, BKS (D)-Leprdb / J, stock number 010803, was purchased from Jax Labs. Bbs10 knockout mice were generated at the Institute Mouse Clinic (ICS) in Strasbourg and have been described in the literature. All mice were on a C57 / BL6 genetic background. All animals were housed in a temperature- and humidity-controlled facility with a 12-h light / 12-h dark cycle. BKS (D)-Leprdb / J mice were fed a chow diet (LM-485; Harlan Teklad Premier Laboratory Diets), while Bbs knockout mice had free access to a high-fat / glucose diet and tap water. For ipGTT and ipITT, mice were fasted for 6 h prior to the start of the experiment. Insulin 0.75 U / kg was injected intravenously via the tail vein. Blood glucose and samples were collected from the tail. Mice were sacrificed by cervical dislocation.

[0184] For the non-obese STAM mouse model study, C57BL / 6 mice (14-day-old pregnant females) were obtained from Japan SLC, Inc. (Japan). All animals used in this study were housed and managed in accordance with the Japanese Pharmacological Society's Guidelines for Animal Use. The animals were housed in an SPF facility under controlled conditions of temperature (23 ± 2°C), humidity (45 ± 10%), lighting (12-h artificial light-dark cycle, light period 8:00–20:00), and air exchange. The animals were housed in TPX cages (CLEA Japan) with a maximum of three mice per cage. Sterilized Paper-Clean (Japan SLC) bedding was used and changed once a week. The cages were covered with a metal lid and the mice were given a sterilized 60% high-fat diet ad libitum. Pure water was provided ad libitum from a water bottle equipped with a rubber stopper and a straw tube. The water bottle was changed once a week, washed, sterilized in an autoclave, and reused.

[0185] Mice were identified by ear punching. Each cage was given a specific identification code. NASH was induced in 12 male mice by a single subcutaneous injection of 200 μg of streptozotocin (STZ, Sigma-Aldrich, USA) solution on day 2 of life and feeding them a high-fat diet (HFD, 57 kcal% fat, Cat. No. HFD32, CLEA Japan, Inc., Japan) from 4 weeks of age onwards.

[0186] Peptide dosing schedule for BKS (D)-Leprdb / J, stock number 010803 and Bbs10 knockout mice Day 0: All mice were fasted for 4 hours in the morning. At 1 pm, control mice were injected once (retroperitoneal fat / subcutaneous injection) with 10 milliliters of 22% glucose solution (grams of body weight). Treated mice were injected with either 25 μg of test peptide (2.5 μL of mother solution) dissolved in 10 milliliters of 22% glucose solution (grams of body weight). Blood glucose was measured in tail vein blood every 30 minutes and plotted to determine the effect of each treatment on the area under the curve.

[0187] Peptide dosing regimen in the STAM mouse model. The test peptide was the ADPIF / PATAS peptide as described above.

[0188] Drug Administration Route The peptides were administered subcutaneously into the adipose tissue in a volume of 100 mL per mouse.

[0189] Experimental design and processing Study Group Group 1: ADPIF / PATAS peptides Six NASH mice were administered vehicle containing ADPIF / PATAS peptide at a dose of 25 μg per mouse subcutaneously into the adipose tissue once a week from 4 to 9 weeks of age.

[0190] Group 2: Vehicle Six NASH mice were administered vehicle (DMSO dissolved in saline) subcutaneously into the adipose tissue once a week from 4 to 9 weeks of age at a volume of 100 mL per mouse.

[0191] The table below summarizes the processing schedule.

[0192] [Table 1]

[0193] FATP expression levels Total RNA was prepared from different tissues and cells using the RiboPure™ kit (catalog number: AM1924; Ambion) followed by DNAse treatment using TURBO DNA-free™ (catalog number: AM1907; Ambion). RNA integrity was assessed by gel electrophoresis and RNA concentration was assessed using an Eppendorf Biophotometer Plus and a Hellma® Tray Cell (catalog number: 105.810-uvs; Hellma). Reverse transcription of 1 μg of total RNA to cDNA was performed using the BioRadiScript™ cDNA Synthesis Kit (catalog number: 170-8891; BioRad). Real-time quantitative polymerase chain reaction amplification was performed in a BioRad CFX96™ Real-Time System using iQ™ SYBR® Green Supermix (Cat. No. 170-8886; BioRad) and target-optimized primer sets validated for SYBR Green-based real-time PCR for real-time PCR. For human primers, all qPCR primers used were purchased from BioRad's validated MIQE primer sets.

[0194] [Table 2]

[0195] Plasma biochemistry measurements Sample collection Plasma samples were collected and stored at -80°C for analysis. The plasma from these mice was then used to measure glycated albumin levels using a commercially available kit from LS Bio, Mouse Glycated Albumin ELISA Kit (Sandwich ELISA)-LS-F28697, Strasbourg.

[0196] Statistical Tests Statistical analysis was performed using GraphPad Prism 6 (GraphPad Software Inc., USA) with Student's t-test. A P value of <0.05 was considered statistically significant. A one-tailed t-test with a P value of <0.1 was considered to indicate a trend. Results were expressed as mean ± SD.

Claims

1. A pharmaceutical composition for the treatment of diabetes and related disorders selected from the group consisting of type I diabetes, type II diabetes, insulin resistance, diabetic retinopathy, diabetic neuropathy, diabetic nephropathy, hyperglycemia, hyperinsulinemia and Bardet-Biedl syndrome, comprising a peptide, - the peptide can reduce the expression of FATP2 in adipose tissue, the peptide does not simultaneously contain one methionine, one proline and one arginine; - the peptide adopts a secondary structure which is a helix, preferably an alpha-helical - the peptide has a length of 12 to 60 amino acids, - the peptide is modified by chemical cross-linking; - the peptide sequence is the following sequence: 【Chemistry 1】 including one of In the formula, the bold and underlined 【Chemistry 2】 the residues of which comprise the staple and are any amino acid derivative suitable for stapling; X is any amino acid except M, P and R.

2. The pharmaceutical composition of claim 1 , wherein the peptide has a length of less than 30 amino acids.

3. The peptide sequence is: 【Chemistry 3】 Containing or consisting solely of In the formula, the bold and underlined 【Chemistry 4】 the residues of which comprise the staple and are any amino acid derivative suitable for stapling; 2. The pharmaceutical composition of claim 1, wherein X is any amino acid except M, P, and R.

4. 4. The pharmaceutical composition according to any one of claims 1 to 3, wherein X is an amino acid which favours an alpha helical secondary structure, or an amino acid selected from the group consisting of A, D, N, C, G, Q, E, H, L, K, F, S, W and Y, or an amino acid selected from the group consisting of A, D, N, G, Q, E, H, L, K, F, S, W and Y.

5. The first bolded and underlined X is R and the second bolded and underlined X is S; 【Chemistry 5】 The pharmaceutical composition according to claim 1 , wherein the pharmaceutical composition comprises a staple.

6. The peptide sequence is 【Chemistry 6】 Including, During the ceremony, 【Chemistry 7】 has a staple, 2. The pharmaceutical composition of claim 1, wherein the peptide optionally further comprises 1 to 5 or 1 to 3 amino acid modifications selected from substitutions, deletions, additions, and combinations thereof.

7. 6. The pharmaceutical composition of claim 5, wherein the peptide comprises 2-(7-octenyl)arginine and 2-(4-pentenyl)serine.

8. 2. The pharmaceutical composition of claim 1, wherein the peptide is used in combination with one or more additional active agents, preferably selected from the group consisting of antidiabetic agents, lipid lowering agents, antiobesity agents, antihypertensive agents, antilipidemic agents, anti-inflammatory agents, and agonists of peroxisome proliferator-activated receptors.

9. 2. The pharmaceutical composition of claim 1, wherein the associated disorder is selected from the group consisting of insulin resistance and Bardet-Biedl syndrome.

Images