IC50: 19.59 nM/L (insulin receptor)[2]

With an IC50 value of 19.59 nM, insulin degludec (0.001-1000 nM; 12 h) binds to the insulin receptor[2]. In HL-1 cells, insulin degludec (200 nM; 10 min) boosts glucose uptake[2].

Insulin degludec (5 U/kg; sc once daily for 30 days) affects glucose homeostasis and liver metabolism in diabetic mice undergoing insulin-induced hypoglycemia[1].

Western Blot Analysis [2]
Cell Types: HL-1 cardiomyocytes
Tested Concentrations: 200 nM
Incubation Duration: 0-60 min
Experimental Results: diminished the level of Akt phosphorylation after 5 and 10 min treatment.

Animal/Disease Models: Male Swiss mice with diabetes[1]
Doses: 5 U/kg
Route of Administration: subcutaneous (sc) injection; 5 U/kg one time/day for 30 days
Experimental Results: demonstrated a fast response to insulin-induced hypoglycemia with a glycemic level at or slightly under 100 mg/dl after 60 min and this response effect can be abolished by cortisol. Diminished rates of glucose production and demonstrated a low lactate production in livers. Increased the number of hepatocytes.

Absorption, Distribution and Excretion
In patients with type 1 diabetes, after 8 consecutive days of once-daily subcutaneous injection of 0.4 U/kg insulin, the median insulin degludec concentration reached a peak of 4472 pmol/L at 9 hours. The median time to onset of action after the first dose was approximately 1 hour. The hypoglycemic effect lasted at least 42 hours after 8 once-daily injections. Steady-state insulin degludec concentrations were reached after 3–4 days. 30% to 80% of circulating insulin is cleared by the kidneys. Using a one-compartment pharmacokinetic model, the apparent volume of distribution was estimated to be 10.6 L in children and 13.9 L in adults. The mean apparent clearance of insulin degludec after a single subcutaneous injection of 0.4 U/kg insulin was 0.03 L/kg (2.1 L/h for a 70 kg individual).

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Metabolism / Metabolites
All metabolites of insulin degludec are inactive. The liver and kidneys play major roles in insulin metabolism. However, although the liver primarily metabolizes endogenous insulin, exogenous insulin is mainly metabolized by the kidneys because it does not directly enter the portal venous system.

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Biological Half-Life
The half-life after subcutaneous administration depends primarily on the absorption rate of the subcutaneous tissue. On average, the steady-state half-life is approximately 25 hours, regardless of the dose.

Protein Binding
Insulin degludec's affinity for serum albumin results in a protein binding rate of over 99% in human plasma. In vitro protein binding studies have shown no clinically significant interactions between insulin degludec and other protein-bound drugs.

[1]. Insulin degludec and glutamine dipeptide modify glucose homeostasis and liver metabolism in diabetic mice undergoing insulin-induced hypoglycemia. J Appl Biomed. 2021 Dec;19(4):210-219.

[2]. Effect of the long-acting insulin analogues glargine and degludec on cardiomyocyte cell signalling and function. Cardiovasc Diabetol. 2016 Jul.

Insulin Degludec is an ultra-long-acting insulin used to treat hyperglycemia caused by type 1 and type 2 diabetes. Insulin is commonly used to treat diabetes, and its mechanism of action mimics the activity of endogenous human insulin. Human insulin is a peptide hormone secreted by pancreatic beta cells that promotes glucose metabolism. After a meal, insulin is released by the pancreas, promoting the absorption of glucose in the blood by visceral organs and tissues such as the liver, fat cells, and skeletal muscle. After being absorbed by cells, glucose is converted into glycogen or fat and stored. In addition, insulin has multiple functions, including inhibiting hepatic glucose production, promoting protein synthesis, and inhibiting fat and protein breakdown. Insulin is a key drug in the treatment of type 1 diabetes (T1D). Type 1 diabetes is caused by an autoimmune response that damages pancreatic beta cells, preventing the body from producing or synthesizing the insulin needed to maintain blood glucose levels. Therefore, patients with type 1 diabetes mainly rely on exogenous insulin, such as degludec, to lower blood glucose levels. Insulin is also used to treat type 2 diabetes (T2D). Type 2 diabetes is another type of diabetes, a slowly progressing metabolic disorder caused by a combination of genetic and lifestyle factors that lead to persistently elevated blood sugar levels. Without treatment or non-pharmacological interventions such as diet and exercise to lower blood sugar, high blood sugar eventually causes cells to become resistant to endogenous insulin, damaging pancreatic cells in the long term. Insulin is usually prescribed later in the course of type 2 diabetes, after trying multiple oral medications (such as [DB00331], [DB01120], or [DB01261]). By this time, pancreatic cells have been sufficiently damaged to prevent the body from producing insulin on its own. Insulin degludec (brand name Tresiba) has a duration of action of up to 42 hours, allowing for once-daily administration, usually injected at bedtime. Due to its long duration of action, Tresiba is considered "basal insulin" because it provides a low concentration of basal insulin, thus maintaining stable blood sugar levels between meals or at night. Basal insulin is typically used in combination with short-acting "pre-meal insulin" (e.g., [DB00046], [DB01309], or [DB01306]) to provide the high dose of insulin required after meals. The combined use of basal and pre-meal insulin is designed to mimic the pancreas's own secretion of endogenous insulin to avoid hypoglycemia. Compared to endogenous insulin, degludec insulin has a hexadecanoic acid group added to the lysine residue at position B29, enabling it to form polymers. After subcutaneous injection, these polymers form a drug reservoir from which monomers are slowly and continuously absorbed into the bloodstream. Therefore, degludec insulin has a longer duration of action due to the slower absorption of the drug from the subcutaneous tissue reservoir into the systemic circulation. Compared to existing long-acting insulin analogs with a duration of action of 20–24 hours (e.g., [DB00047] and [DB01307]), degludec insulin provides stable basal insulin levels over 42 hours with a low peak-to-trough ratio. Limitations of short-acting insulin analogs include more frequent dosing and poorer pharmacokinetic stability, which can negatively impact patient adherence and glycemic control, especially at night. Without sufficient insulin to promote glucose uptake in the blood, blood glucose levels can rise dangerously high, leading to symptoms such as fatigue, headache, blurred vision, and increased thirst. If left untreated, the body will begin breaking down fat instead of glucose for energy, causing ketoacidosis, a life-threatening medical emergency. In the long term, high blood sugar increases the risk of heart attack, stroke, and diabetic neuropathy. Insulin degludec, marketed as Tresiba, was approved by the U.S. Food and Drug Administration (FDA) in September 2015 for the treatment of glycemic control in adults with diabetes. Insulin degludec is an insulin analog. It is a recombinant long-acting human insulin analog with glycemic-lowering properties. After subcutaneous injection, insulin degludec forms a hexamethylenetetramer reservoir from which insulin monomers are slowly released into the systemic circulation. Insulin regulates glucose metabolism by binding to insulin receptors on muscle and fat cells, thereby stimulating cellular glucose uptake and lowering blood sugar levels. Insulin inhibits the liver's conversion of stored glycogen into glucose, which also helps lower blood sugar levels. Furthermore, insulin inhibits lipolysis in adipose tissue, inhibits protein hydrolysis, and promotes protein synthesis. Insulin degludec does not contain threonine (the amino acid at position B30) of human insulin; in addition, lysine (position B29) binds to hexadecanoic acid, which allows for the formation of a multimeric hexamer after subcutaneous injection. Phenol and zinc present in the formulation promote the formation of both dimers and hexamers in insulin degludec.
See also: Insulin degludec; Liraglutide (ingredient); Insulin aspart; Insulin degludec (ingredient).

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Drug Indication
Insulin degludec is indicated for improving glycemic control in patients with diabetes aged 1 year and older.
FDA label

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Mechanism of Action
Insulin Degludec is a heterotetrameric protein composed of two extracellular α subunits and two transmembrane β subunits. Upon binding to the α subunit of the insulin receptor (IR), it stimulates the intrinsic tyrosine kinase activity of the receptor's β subunit. The bound receptor is autophosphorylated, leading to phosphorylation of various intracellular substrates, such as insulin receptor substrate (IRS) proteins, Cbl, APS, Shc, and Gab 1. Activation of these proteins results in the activation of downstream signaling molecules, including PI3 kinase and Akt. Akt regulates the activity of glucose transporter 4 (GLUT4) and protein kinase C (PKC), both of which play crucial roles in both metabolism and catabolism.

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Pharmacodynamics
Insulin Degludec is a natural hormone secreted by the beta cells of the pancreas. In non-diabetic individuals, the pancreas continuously secretes low levels of basal insulin, with a postprandial insulin spike. This increased postprandial insulin secretion is due to the metabolic changes that occur as the body transitions from a postabsorbent to an absorptive state. Insulin promotes cellular glucose uptake, particularly in muscle and adipose tissue; promotes energy storage through glycogen synthesis; inhibits catabolism of energy reserves; increases DNA replication and protein synthesis by stimulating amino acid uptake in the liver, muscle, and adipose tissue; and regulates the activity of various enzymes involved in glycogen synthesis and glycolysis. Insulin also promotes growth and is essential for the functioning of growth hormone (e.g., protein synthesis, cell division, DNA synthesis). Detemir insulin is a long-acting insulin analog with a smooth and predictable action curve. It mimics the basal insulin levels of diabetic patients. Detemir insulin has an onset time of 1 to 2 hours and a duration of action of up to 24 hours. Notably, its affinity for the insulin receptor (30%) is lower than that of human insulin.

6101.843

844439-96-9

118984462

White to off-white solid powder

79

92

197

426

15300

51

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FYZPCMFQCNBYCY-WIWKJPBBSA-N

InChI=1S/C274H411N65O81S6/c1-28-146(23)223(331-210(357)119-275)268(412)335-222(145(21)22)264(408)305-172(89-97-218(369)370)234(378)300-169(84-92-204(279)351)238(382)326-198-131-424-425-132-199-259(403)323-193(126-341)256(400)312-177(103-138(7)8)241(385)313-183(110-154-66-74-160(345)75-67-154)244(388)301-167(82-90-202(277)349)235(379)308-176(102-137(5)6)239(383)303-171(88-96-217(367)368)237(381)320-189(116-205(280)352)251(395)316-185(112-156-70-78-162(347)79-71-156)247(391)327-197(258(402)322-191(273(419)420)118-207(282)354)130-423-422-129-196(231(375)290-122-211(358)295-166(86-94-215(363)364)233(377)299-165(62-53-99-288-274(283)284)229(373)289-123-212(359)297-181(108-152-57-45-41-46-58-152)243(387)315-182(109-153-59-47-42-48-60-153)246(390)317-186(113-157-72-80-163(348)81-73-157)254(398)338-226(150(27)344)270(414)339-100-54-63-201(339)262(406)306-173(271(415)416)61-51-52-98-287-208(355)93-85-174(272(417)418)296-209(356)64-49-38-36-34-32-30-31-33-35-37-39-50-65-214(361)362)329-266(410)221(144(19)20)334-252(396)179(105-140(11)12)310-245(389)184(111-155-68-76-161(346)77-69-155)314-240(384)175(101-136(3)4)307-227(371)148(25)294-232(376)170(87-95-216(365)366)304-263(407)220(143(17)18)333-253(397)180(106-141(13)14)311-249(393)188(115-159-121-286-135-293-159)319-255(399)192(125-340)298-213(360)124-291-230(374)195(128-421-426-133-200(328-260(198)404)261(405)337-225(149(26)343)269(413)324-194(127-342)257(401)336-224(147(24)29-2)267(411)330-199)325-242(386)178(104-139(9)10)309-248(392)187(114-158-120-285-134-292-158)318-236(380)168(83-91-203(278)350)302-250(394)190(117-206(281)353)321-265(409)219(142(15)16)332-228(372)164(276)107-151-55-43-40-44-56-151/h40-48,55-60,66-81,120-121,134-150,164-201,219-226,340-348H,28-39,49-54,61-65,82-119,122-133,275-276H2,1-27H3,(H2,277,349)(H2,278,350)(H2,279,351)(H2,280,352)(H2,281,353)(H2,282,354)(H,285,292)(H,286,293)(H,287,355)(H,289,373)(H,290,375)(H,291,374)(H,294,376)(H,295,358)(H,296,356)(H,297,359)(H,298,360)(H,299,377)(H,300,378)(H,301,388)(H,302,394)(H,303,383)(H,304,407)(H,305,408)(H,306,406)(H,307,371)(H,308,379)(H,309,392)(H,310,389)(H,311,393)(H,312,400)(H,313,385)(H,314,384)(H,315,387)(H,316,395)(H,317,390)(H,318,380)(H,319,399)(H,320,381)(H,321,409)(H,322,402)(H,323,403)(H,324,413)(H,325,386)(H,326,382)(H,327,391)(H,328,404)(H,329,410)(H,330,411)(H,331,357)(H,332,372)(H,333,397)(H,334,396)(H,335,412)(H,336,401)(H,337,405)(H,338,398)(H,361,362)(H,363,364)(H,365,366)(H,367,368)(H,369,370)(H,415,416)(H,417,418)(H,419,420)(H4,283,284,288)/t146-,147-,148-,149+,150+,164-,165-,166-,167-,168-,169-,170-,171-,172-,173-,174-,175-,176-,177-,178-,179-,180-,181-,182-,183-,184-,185-,186-,187-,188-,189-,190-,191-,192-,193-,194-,195-,196-,197-,198-,199-,200-,201-,219-,220-,221-,222-,223-,224-,225-,226-/m0/s1

16-[[(1S)-4-[[(5S)-5-[[(2S)-1-[(2S,3R)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[2-[[(2S)-2-[[(2S)-2-[[2-[[(1R,6R,12S,15S,18S,21S,24S,27S,30S,33S,36S,39S,42R,47R,50S,53S,56S,59S,62S,65S,68S,71S,74R,77S,80S,83S,88R)-88-[[(2S)-5-amino-2-[[(2S)-2-[[(2S)-2-[[(2S,3S)-2-[(2-aminoacetyl)amino]-3-methylpentanoyl]amino]-3-methylbutanoyl]amino]-4-carboxybutanoyl]amino]-5-oxopentanoyl]amino]-6-[[(2S)-2-[[(2S)-2-[[(2S)-5-amino-2-[[(2S)-4-amino-2-[[(2S)-2-[[(2S)-2-amino-3-phenylpropanoyl]amino]-3-methylbutanoyl]amino]-4-oxobutanoyl]amino]-5-oxopentanoyl]amino]-3-(1H-imidazol-4-yl)propanoyl]amino]-4-methylpentanoyl]amino]-47-[[(1S)-3-amino-1-carboxy-3-oxopropyl]carbamoyl]-53-(2-amino-2-oxoethyl)-62-(3-amino-3-oxopropyl)-77-[(2S)-butan-2-yl]-24,56-bis(2-carboxyethyl)-83-[(1R)-1-hydroxyethyl]-12,71,80-tris(hydroxymethyl)-33,50,65-tris[(4-hydroxyphenyl)methyl]-15-(1H-imidazol-4-ylmethyl)-27-methyl-18,30,36,59,68-pentakis(2-methylpropyl)-7,10,13,16,19,22,25,28,31,34,37,40,49,52,55,58,61,64,67,70,73,76,79,82,85,87-hexacosaoxo-21,39-di(propan-2-yl)-3,4,44,45,90,91-hexathia-8,11,14,17,20,23,26,29,32,35,38,41,48,51,54,57,60,63,66,69,72,75,78,81,84,86-hexacosazabicyclo[72.11.7]dononacontane-42-carbonyl]amino]acetyl]amino]-4-carboxybutanoyl]amino]-5-carbamimidamidopentanoyl]amino]acetyl]amino]-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-3-(4-hydroxyphenyl)propanoyl]amino]-3-hydroxybutanoyl]pyrrolidine-2-carbonyl]amino]-5-carboxypentyl]amino]-1-carboxy-4-oxobutyl]amino]-16-oxohexadecanoic acid

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

0.1 M HCL: 50 mg/mL

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)