Insulin Detemir targets the insulin receptor (IR). As a long-acting insulin analogue, it binds to the insulin receptor and activates downstream signaling pathways including Akt and ERK. Activation of these pathways promotes glucose uptake, glycogen synthesis, and GLP-1 secretion. The compound is designed to have a prolonged duration of action through albumin binding, which slows its clearance from the circulation.

Insulin Detemir (d-INS) (100 nM; 0.5-4 h) enhances Gcg mRNA expression in primary fetal rat intestinal cell (FRIC) cultures, and (100 nM; 5 min and 10 min) promotes fast phosphorylation of Akt, as well [1]. Insulin Detemir (100 nM; 5-120 min) increases β-catenin phosphorylation, its nuclear translocation, and enhances cAMP response element-binding protein (CREB) phosphorylation in a phosphatidylinositol 3-kinase and/or mitogen-activated protein kinase kinase/extracellular signal-regulated kinase-sensitive manner[1].

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Insulin Detemir demonstrates in vitro activity by binding to the insulin receptor and activating Akt- and ERK-dependent signaling pathways. It promotes GLP-1 secretion by enhancing glucagon gene expression in vitro. The compound effectively regulates glucose metabolism in cell-based assays by promoting glucose uptake and glycogen synthesis.

Insulin Detemir (d-INS) (5 IU/kg; ip; once daily; 2 weeks) exhibits a preference for intestinal tissues and, in comparison to other insulin formulations, has weight-sparing benefits. This may be due to the activation of the insulin/-catenin/CREB signaling pathways.

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Insulin Detemir exhibits potent in vivo activity as a long-acting insulin analogue for the treatment of diabetes. It effectively regulates blood glucose levels in patients with Type 1 and Type 2 Diabetes. The compound has a prolonged duration of effect due to albumin binding, allowing for once- or twice-daily dosing. It has been widely used in clinical practice for the management of hyperglycemia.

The in vitro receptor binding assay for Insulin Detemir involves assessing its binding affinity to the insulin receptor. Radioligand binding assays are performed using membrane preparations or cells expressing the insulin receptor. The compound is incubated with a labeled insulin receptor ligand, and displacement of the labeled ligand is measured to determine binding affinity. Receptor activation is assessed by measuring downstream signaling such as Akt and ERK phosphorylation.

Western Blot Analysis[1]
Cell Types: GLUTag cells
Tested Concentrations: 100 nM
Incubation Duration: 0, 5, 10, 30, 60, and 120 min
Experimental Results: Stimulated CREB , ERK1/2, Akt and its downstream glycogen synthase kinase (GSK)-3 phosphorylation at 5 min and 10 min.

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The in vitro cell-based assay for Insulin Detemir involves treating cells expressing the insulin receptor with the compound and measuring downstream signaling and functional responses. Cells are treated with Insulin Detemir, and activation of Akt and ERK pathways is assessed by Western blotting with phospho-specific antibodies. Glucose uptake, glycogen synthesis, and GLP-1 secretion are also measured to evaluate the compound’s metabolic effects.

Animal/Disease Models: Obese type 2 diabetic db/db mice[1]
Doses: 5 IU/kg
Route of Administration: intraperitoneal (ip) injection; one time/day for 2 weeks
Experimental Results: diminished body weight of the mice after 14-day daily injection of d-INS (5 IU/kg) Dramatically compared with those injected with the same dose of human Insulin or saline. Induced rapid phosphorylation of protein kinase B (Akt) in the gut L cells of normal mice.

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In vivo animal experiments for Insulin Detemir are conducted in diabetic animal models to evaluate its glucose-lowering efficacy. Diabetic animals are administered Insulin Detemir via subcutaneous injection, and blood glucose levels are monitored over time. The compound’s duration of action, glucose-lowering effect, and effects on GLP-1 secretion are assessed.

Absorption, Distribution and Excretion
In healthy subjects and diabetic patients, serum concentrations of insulin detemir after subcutaneous injection exhibited a relatively stable concentration-time curve over 24 hours, with the maximum serum concentration (Cmax) reached 6–8 hours post-administration. In adult patients with type 1 diabetes, a single administration of 0.5 units/kg of insulin detemir resulted in a maximum serum concentration (Cmax) of 4,641 ± 2,299 pmol/L. Subcutaneous injection into the thigh resulted in slower absorption of insulin detemir, with its AUC0–5h being 30%–40% lower than the corresponding AUCs after subcutaneous injection into the deltoid and abdomen, and 10% lower from AUC0–∞. Insulin detemir absorption is slow and prolonged, with a relatively stable concentration-time curve over 24 hours without a significant peak. The median time to reach maximum serum insulin concentration was 12 hours post-injection. On average, serum insulin concentrations decreased to baseline levels within approximately 24 hours. The absolute bioavailability of insulin detemir is approximately 60%. 30%–80% of circulating insulin is cleared by the kidneys. The apparent volume of distribution of insulin detemir is approximately 0.1 L/kg. The apparent clearance rate (CL/F) is fairly consistent across different patient populations with type 1 diabetes. It is estimated that the CL/F is 3.43 ± 1.36 L/min·kg for patients aged 6 to 12 years, 3.74 ± 0.98 L/min·kg for patients aged 13 to 17 years, and 3.41 ± 1.00 L/min·kg for adults (18 to 65 years).

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Metabolism / Metabolites
The liver and kidneys play major roles in insulin metabolism. However, while the liver primarily metabolizes endogenous insulin, exogenous insulin is mainly metabolized by the kidneys because it cannot directly enter the portal venous system.

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Biological Half-Life
After subcutaneous injection of insulin detemir in patients with type 2 diabetes, the terminal half-life is 5 to 7 hours, depending on the dose.

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Insulin Detemir is a long-acting insulin analogue with a prolonged duration of effect due to albumin binding. It has a molecular formula of C267H402N64O76S6 and a molecular weight of 5916.9 g/mol. The compound is administered subcutaneously and has a duration of action of up to 24 hours. It is typically dosed once or twice daily for the management of diabetes.

Protein Binding
Over 98% of insulin detemir in the blood is bound to albumin. In vitro and in vivo protein binding studies have shown no clinically significant interactions between insulin detemir and fatty acids or other protein-bound drugs.

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Insulin Detemir has been extensively studied in clinical trials and is approved for the treatment of diabetes. Common adverse effects include hypoglycemia, injection site reactions, and weight gain. The safety profile is well-established through long-term clinical use. As a therapeutic agent, it is available under the brand name Levemir.

[1]. Insulin detemir enhances proglucagon gene expression in the intestinal L cells via stimulating β-catenin and CREB activities. Am J Physiol Endocrinol Metab. 2012 Sep 15;303(6):E740-51.

recombinant long-acting insulin and hypoglycemic agent, wherein myristic acid is bound to lysine at position B29. It is used to control blood glucose levels in patients with diabetes.
See also: Insulin detemir (note moved to); Recombinant insulin detemir (note moved to).

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Insulin Detemir (CAS 169148-63-4) is a long-acting insulin analogue used for the treatment of Type 1 and Type 2 Diabetes. It has a molecular formula of C267H402N64O76S6 and a molecular weight of 5916.9 g/mol. It is a soluble, basal insulin analogue with a prolonged duration of effect due to albumin binding. It is approved for clinical use and is available under the brand name Levemir. It promotes GLP-1 secretion through Akt-/ERK-dependent signaling.

5914.795

169148-63-4

16137271

Colorless to light yellow liquid

-3.5

76

87

189

413

14700

50

UGOZVNFCFYTPAZ-IOXYNQHNSA-N

InChI=1S/C267H402N64O76S6/c1-29-32-33-34-35-36-37-38-39-40-50-64-204(347)280-95-52-51-61-170(265(404)405)298-256(395)197-63-54-97-331(197)264(403)220(147(28)336)330-248(387)182(110-154-71-79-160(340)80-72-154)309-240(379)178(106-150-59-48-43-49-60-150)307-237(376)177(105-149-57-46-42-47-58-149)289-207(350)120-282-223(362)162(62-53-96-281-267(276)277)291-227(366)163(84-91-209(352)353)288-206(349)119-283-225(364)192-126-409-410-127-193(252(391)314-187(266(406)407)115-203(275)346)319-241(380)181(109-153-69-77-159(339)78-70-153)308-245(384)185(113-201(273)344)312-231(370)168(86-93-211(356)357)295-233(372)172(99-134(6)7)300-229(368)164(81-88-198(270)341)293-238(377)179(107-151-65-73-157(337)74-66-151)305-235(374)173(100-135(8)9)304-250(389)189(123-333)315-253(392)195-129-412-411-128-194(318-232(371)166(83-90-200(272)343)292-228(367)169(87-94-212(358)359)297-258(397)216(142(22)23)327-262(401)217(143(24)30-2)323-205(348)116-268)254(393)320-196(255(394)329-219(146(27)335)263(402)316-190(124-334)251(390)328-218(144(25)31-3)261(400)322-195)130-413-408-125-191(317-236(375)174(101-136(10)11)301-242(381)183(111-155-117-278-131-285-155)310-230(369)165(82-89-199(271)342)294-244(383)186(114-202(274)345)313-259(398)213(139(16)17)324-222(361)161(269)104-148-55-44-41-45-56-148)224(363)284-121-208(351)290-188(122-332)249(388)311-184(112-156-118-279-132-286-156)243(382)303-176(103-138(14)15)247(386)325-214(140(18)19)257(396)296-167(85-92-210(354)355)226(365)287-145(26)221(360)299-171(98-133(4)5)234(373)306-180(108-152-67-75-158(338)76-68-152)239(378)302-175(102-137(12)13)246(385)326-215(141(20)21)260(399)321-192/h41-49,55-60,65-80,117-118,131-147,161-197,213-220,332-340H,29-40,50-54,61-64,81-116,119-130,268-269H2,1-28H3,(H2,270,341)(H2,271,342)(H2,272,343)(H2,273,344)(H2,274,345)(H2,275,346)(H,278,285)(H,279,286)(H,280,347)(H,282,362)(H,283,364)(H,284,363)(H,287,365)(H,288,349)(H,289,350)(H,290,351)(H,291,366)(H,292,367)(H,293,377)(H,294,383)(H,295,372)(H,296,396)(H,297,397)(H,298,395)(H,299,360)(H,300,368)(H,301,381)(H,302,378)(H,303,382)(H,304,389)(H,305,374)(H,306,373)(H,307,376)(H,308,384)(H,309,379)(H,310,369)(H,311,388)(H,312,370)(H,313,398)(H,314,391)(H,315,392)(H,316,402)(H,317,375)(H,318,371)(H,319,380)(H,320,393)(H,321,399)(H,322,400)(H,323,348)(H,324,361)(H,325,386)(H,326,385)(H,327,401)(H,328,390)(H,329,394)(H,330,387)(H,352,353)(H,354,355)(H,356,357)(H,358,359)(H,404,405)(H,406,407)(H4,276,277,281)/t143-,144-,145-,146+,147+,161-,162-,163-,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-,213-,214-,215-,216-,217-,218-,219-,220-/m0/s1

(2S)-2-[[(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]-6-(tetradecanoylamino)hexanoic acid

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

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

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

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.)