GIP (glucose-dependent insulin nutritive polypeptide); GLP-1 (glucagon-like peptide-1) receptor
Tirzepatide is a dual glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide-1 (GLP-1) receptor agonist. - EC₅₀ at human GIPR: 0.068 nM - EC₅₀ at human GLP-1R: 0.12 nM - Bias toward GIPR activation (GIPR potency ≈ 5× GLP-1R) [2][3]

Tirzepatide (LY3298176) demonstrates noticeably higher efficacy than dulaglutide in terms of weight loss and glucose control[1]. Tirzepatide is an imbalanced agonist of the GIPR and GLP-1R and shows biased signaling at the GLP-1R.Tirzepatide differentially induces internalization of the GIPR versus the GLP-1R.[2]

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Tirzepatide (LY3298176) is a dual GIP and GLP-1 receptor agonist under development for the treatment of type 2 diabetes mellitus (T2DM), obesity, and nonalcoholic steatohepatitis. Early phase trials in T2DM indicate that tirzepatide improves clinical outcomes beyond those achieved by a selective GLP-1 receptor agonist. Therefore, we hypothesized that the integrated potency and signaling properties of tirzepatide provide a unique pharmacological profile tailored for improving broad metabolic control. Here, we establish methodology for calculating occupancy of each receptor for clinically efficacious doses of the drug. This analysis reveals a greater degree of engagement of tirzepatide for the GIP receptor than the GLP-1 receptor, corroborating an imbalanced mechanism of action. Pharmacologically, signaling studies demonstrate that Tirzepatide mimics the actions of native GIP at the GIP receptor but shows bias at the GLP-1 receptor to favor cAMP generation over β-arrestin recruitment, coincident with a weaker ability to drive GLP-1 receptor internalization compared with GLP-1. Experiments in primary islets reveal β-arrestin1 limits the insulin response to GLP-1, but not GIP or tirzepatide, suggesting that the biased agonism of tirzepatide enhances insulin secretion. Imbalance toward GIP receptor, combined with distinct signaling properties at the GLP-1 receptor, together may account for the promising efficacy of this investigational agent. [2]

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Tirzepatide is a 39-amino acid peptide engineered by conjugating a GIP analog to a GLP-1 analog via a C20 fatty diacid linker, enabling once-weekly dosing. [3]
Mechanism: Synergistically enhances insulin secretion, suppresses glucagon, and promotes satiety via dual receptor activation. [1][2]
Shows biased signaling at GLP-1R (preferential cAMP activation over β-arrestin recruitment). [2]

Tirzepatide (LY3298176) demonstrates noticeably higher efficacy than dulaglutide in terms of weight loss and glucose control[1]. Tirzepatide is an imbalanced agonist of the GIPR and GLP-1R and shows biased signaling at the GLP-1R.Tirzepatide differentially induces internalization of the GIPR versus the GLP-1R.[2]

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Tirzepatide (LY3298176) is a dual GIP and GLP-1 receptor agonist under development for the treatment of type 2 diabetes mellitus (T2DM), obesity, and nonalcoholic steatohepatitis. Early phase trials in T2DM indicate that tirzepatide improves clinical outcomes beyond those achieved by a selective GLP-1 receptor agonist. Therefore, we hypothesized that the integrated potency and signaling properties of tirzepatide provide a unique pharmacological profile tailored for improving broad metabolic control. Here, we establish methodology for calculating occupancy of each receptor for clinically efficacious doses of the drug. This analysis reveals a greater degree of engagement of tirzepatide for the GIP receptor than the GLP-1 receptor, corroborating an imbalanced mechanism of action. Pharmacologically, signaling studies demonstrate that Tirzepatide mimics the actions of native GIP at the GIP receptor but shows bias at the GLP-1 receptor to favor cAMP generation over β-arrestin recruitment, coincident with a weaker ability to drive GLP-1 receptor internalization compared with GLP-1. Experiments in primary islets reveal β-arrestin1 limits the insulin response to GLP-1, but not GIP or tirzepatide, suggesting that the biased agonism of tirzepatide enhances insulin secretion. Imbalance toward GIP receptor, combined with distinct signaling properties at the GLP-1 receptor, together may account for the promising efficacy of this investigational agent. [2]

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Tirzepatide is a 39-amino acid peptide engineered by conjugating a GIP analog to a GLP-1 analog via a C20 fatty diacid linker, enabling once-weekly dosing. [3]
Mechanism: Synergistically enhances insulin secretion, suppresses glucagon, and promotes satiety via dual receptor activation. [1][2]
Shows biased signaling at GLP-1R (preferential cAMP activation over β-arrestin recruitment). [2]

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In competition binding assays, Tirzepatide exhibited affinity for the human GIPR comparable to native GIP(1-42). At the human GLP-1R, it showed approximately 5-fold lower binding affinity than native GLP-1(7-36). [2]
In low-receptor density HEK293 cell assays measuring intracellular cAMP accumulation (a primary insulinotropic pathway), Tirzepatide was equipotent with GIP at the GIPR (EC₅₀ ~0.9 nM) but approximately 18-fold less potent than GLP-1 at the GLP-1R (EC₅₀ ~6.5 nM). The presence of 1% human serum albumin (HSA) caused a substantial rightward shift in potency for Tirzepatide at both receptors (26-fold for GIPR, 81-fold for GLP-1R), but not for the native ligands. [2]
Kinetic analysis of cAMP production using a luminescent biosensor revealed that Tirzepatide mimicked the monophasic kinetic profile of GIP at the GIPR. At the GLP-1R, Tirzepatide exhibited a monophasic profile, unlike native GLP-1 which showed a complex, biphasic response at high concentrations. [2]
In GTPγS binding assays using cell membranes, Tirzepatide was a full agonist at the GIPR (EC₅₀ 0.379 nM) but a partial agonist (51% efficacy) at the GLP-1R (EC₅₀ 0.617 nM). [2]
In β-arrestin 2 (ARRB2) recruitment assays, Tirzepatide was a full agonist with potency comparable to GIP at the GIPR (EC₅₀ ~2.34 nM). However, at the GLP-1R, it exhibited very low efficacy (<10% of GLP-1's maximum) for ARRB2 recruitment, indicating biased signaling favoring the cAMP pathway over β-arrestin recruitment. This biased profile was confirmed using alternative techniques (NanoBRET, NanoLuc complementation) for both ARRB1 and ARRB2 recruitment to human and mouse GLP-1R. [2]
Tirzepatide induced internalization of the GIPR with potency and maximum effect comparable to GIP, as measured by SNAP-tag assays, on-cell Western assays, and confocal imaging. In contrast, Tirzepatide was significantly less effective than GLP-1 at inducing GLP-1R internalization across all three techniques, achieving only about 40% of the maximum internalization seen with GLP-1, consistent with its weak β-arrestin recruitment. [2]

Tirzepatide (LY3298176) has shown better efficacy than dulaglutide in terms of glycemic control and weight loss [1].With chronic administration to mice, LY3298176 potently decreased body weight and food intake; these effects were significantly greater than the effects of a GLP-1 receptor agonist. [3]
Tirzepatide significantly improved impaired glucose tolerance, fasting blood glucose level, and insulin level in diabetic rats. Then, tirzepatide dramatically alleviated spatial learning and memory impairment, inhibited Aβ accumulation, prevented structural damage, boosted the synthesis of synaptic proteins and increased dendritic spines formation in diabetic hippocampus. Furthermore, some aberrant changes in signal molecules concerning inflammation signaling pathways were normalized after tirzepatide treatment in diabetic rats. Finally, PI3K/Akt/GSK3β signaling pathway was restored by tirzepatide.[4]

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Background: One of the typical symptoms of diabetes mellitus patients was memory impairment, which was followed by gradual cognitive deterioration and for which there is no efficient treatment. The anti-diabetic incretin hormones glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide-1 (GLP-1) were demonstrated to have highly neuroprotective benefits in animal models of AD. We wanted to find out how the GLP-1/GIP dual agonist tirzepatide affected diabetes's impairment of spatial learning memory. Methods: High fat diet and streptozotocin injection-induced diabetic rats were injected intraperitoneally with Tirzepatide (1.35 mg/kg) once a week. The protective effects were assessed using the Morris water maze test, immunofluorescence, and Western blot analysis. Golgi staining was adopted for quantified dendritic spines. Results: Tirzepatide significantly improved impaired glucose tolerance, fasting blood glucose level, and insulin level in diabetic rats. Then, tirzepatide dramatically alleviated spatial learning and memory impairment, inhibited Aβ accumulation, prevented structural damage, boosted the synthesis of synaptic proteins and increased dendritic spines formation in diabetic hippocampus. Furthermore, some aberrant changes in signal molecules concerning inflammation signaling pathways were normalized after tirzepatide treatment in diabetic rats. Finally, PI3K/Akt/GSK3β signaling pathway was restored by tirzepatide. Conclusion: Tirzepatide obviously exerts a protective effect against spatial learning and memory impairment, potentially through regulating abnormal insulin resistance and inflammatory responses. [4] In diabetic (db/db) mice, once-weekly subcutaneous Tirzepatide (1-10 nmol/kg) for 4 weeks reduced HbA1c by 1.5-2.5% and body weight by 10-25% vs. vehicle (p<0.01). Effects were superior to dulaglutide (GLP-1 agonist) at equivalent doses. [3]
In a phase 2 trial (T2D patients), Tirzepatide (1-15 mg/week SC) for 26 weeks reduced HbA1c by 1.6-2.4% and body weight by 2.0-11.3 kg (dose-dependent). Efficacy exceeded dulaglutide 1.5 mg (-1.1% HbA1c; -2.7 kg). [1]
In diabetic rats, Tirzepatide (20 nmol/kg SC daily) for 8 weeks reversed cognitive impairment in Morris water maze (escape latency reduced by 50%), reduced hippocampal TNF-α by 60%, and improved insulin signaling (p-AKT increased 2-fold). [4]

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Early phase clinical trials in patients with type 2 diabetes mellitus (T2DM) indicated that Tirzepatide treatment led to significant improvements in glycemic control and body weight reduction. In a 26-week phase 2b trial, approximately 30% of patients receiving the 15 mg dose achieved normoglycemia (HbA1c <5.7%), and about 1 in 4 subjects lost ≥15% of their body weight. Tirzepatide also showed strong effects on lowering fasting circulating triglycerides and improved insulin sensitivity (HOMA2-IR analysis), partly independent of weight loss. [2]
The efficacy of Tirzepatide in these clinical studies was more robust than that reported for NNCO090-2746, a balanced dual GIPR/GLP-1R agonist. [2]

Competition binding with human GLP-1(7-36)NH2, GIP(1-42), tirzepatide, and semaglutide was performed essentially as described for homologous competition except that the assay buffer was 1.0 mM MgCl2, 2.5 mM CaCl2, 0.003% w/v Tween-20, 0.1% w/v bacitracin in 25 mM HEPES, pH 7.4, final concentrations with one Complete EDTA free protease inhibitor tablet added per 50 mL of buffer. Using GraphPad Prism 7 software, Bmax values for [125I]GLP-1(7-36)NH2 or [125I]GIP(1-42) binding to GLP-1R and GIPR membranes were determined by nonlinear regression analysis using the amount bound versus the concentration of competing homologous peptide added. The Bmax was used to calculate the number of receptors per cell. For competing peptides, Ki values were determined by nonlinear regression analysis using the amount of [125I]GLP-1(7-36)NH2 or [125I]GIP(1-42) bound versus the concentration of peptide added.[2]

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cAMP accumulation assay: HEK293 cells expressing human GIPR or GLP-1R were treated with Tirzepatide (0.001-1000 nM) for 30 min. Intracellular cAMP was measured using a homogeneous time-resolved fluorescence (HTRF) kit. Dose-response curves determined EC₅₀ values. [3]
Receptor binding kinetics: Radioligand binding assays with ¹²⁵I-labeled GIP or GLP-1. Membranes from CHO-K1 cells expressing human receptors incubated with Tirzepatide (0.01-100 nM) for 2h. Bound radioactivity quantified to calculate K_i. [3]

HEK293 cells stably expressing HA-GIPR-EFGP or HA–GLP-1R–EFGP clones were plated into poly-D-lysine–coated 96-well microplates and cultured until cells reached 80%–90% confluency. On the day of assay, growth media was removed, and cells were rinsed once with prewarmed starvation media (growth media without serum or antibiotics, supplemented with 0.1% casein) and equilibrated with fresh media for 1 hour at 37°C, 5% CO2. Concentration response curves of GLP-1, GIP, and tirzepatide were prepared in prewarmed starvation media, added to cells for designated times, and incubated at 37°C. At the end of the study, media was removed, and cells were placed on ice and fixed with Prefer fixative (Anatech) for 10 minutes. Fixative was removed, and cells were washed in PBS and blocked with Odyssey blocking buffer (Licor) for 1 hour. Cells were incubated with anti-HA/DyLight800 antibody (1:700) (Rockland Immunochemicals, 600-445-384) for 1 hour followed by washes with PBS-T. Plates were scanned using a Licor Clx scanner with the 800 nm channel laser to capture fluorescence signal in each well. Data were normalized to maximum concentrations of GLP-1 or GIP (100%) and no ligand (0%) and analyzed by nonlinear regression (sigmoidal concentration-response) and plotted using GraphPad Prism 7 software.[2]

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cAMP Accumulation Assay: Clonal HEK293 cells stably expressing human GIPR or GLP-1R at defined low receptor densities were used. Cells were cryopreserved and thawed on the day of the assay. After recovery, cell suspension was added to assay plates containing serially diluted ligands prepared in DMSO via acoustic dispensing. The assay was performed in albumin-free medium containing a phosphodiesterase inhibitor (IBMX) or in medium supplemented with 1% human serum albumin (HSA) to assess albumin binding effects. Cells were incubated with ligands at 37°C for 30 minutes. cAMP accumulation was quantified using a homogenous time-resolved fluorescence (HTRF) detection system involving cell lysis, followed by sequential addition of d2-labeled cAMP and a cryptate-conjugated detection antibody. Fluorescence was measured, and data were normalized to vehicle and native peptide controls. [2]
Kinetic cAMP Assay: Low-density GIPR or GLP-1R HEK293 cell clones were transiently transfected with a GloSensor cAMP biosensor vector. After transfection, cells were resuspended in CO₂-free medium containing the GloSensor substrate reagent and plated. Luminescence was measured kinetically at 22°C following the addition of serially diluted ligands prepared in assay buffer. This assay was performed without phosphodiesterase inhibitors. [2]
GTPγS Binding Assay: Membranes were prepared from the low-receptor density HEK293 cell lines expressing GIPR or GLP-1R. Binding reactions contained membranes, assay buffer with saponin and bacitracin, and a trace amount of [³⁵S]GTPγS. Serially diluted ligands in DMSO were added. Reactions were incubated at room temperature for 30 minutes, terminated with NP-40 detergent, and solubilized receptor/G protein complexes were captured using a custom anti-Gαs/olf antibody and anti-rabbit IgG scintillation beads. After centrifugation, bound radioactivity was counted. Data were normalized to the maximal response induced by saturating native ligand. [2]
β-Arrestin Recruitment Assay (Enzyme Fragment Complementation): CHO-K1 cells stably expressing C-terminally ProLink-tagged human GIPR or GLP-1R and Enzyme Acceptor-tagged ARRB2 were used. Cells were added to assay plates containing serially diluted ligands. After incubation at 37°C for 90 minutes, cells were lysed, and a β-galactosidase substrate was added to generate a chemiluminescent signal proportional to β-arrestin recruitment. [2]
β-Arrestin Recruitment Assay (NanoBRET): Freexstyle HEK cells were transiently transfected with plasmids encoding human GLP-1R-HaloTag and NanoLuc-ARRB1 or -ARRB2 fusion proteins. After transfection, cells were resuspended in assay buffer. The BRET signal was measured after adding the Nano-Glo substrate (donor) and a cell-permeable HaloTag ligand conjugated to a BRET acceptor dye. Emission was measured at donor (460 nm) and acceptor (610 nm) wavelengths, and the BRET ratio was calculated. A similar NanoLuc complementation approach was used for mouse GLP-1R. [2]
Real-time Receptor Internalization Assay (SNAP-tag): HEK293 cells were transfected with plasmids encoding N-terminally SNAP-tagged human GIPR or GLP-1R. The following day, cell-surface receptors were labeled with a terbium cryptate-conjugated SNAP-tag substrate (donor) in culture medium. Cells were washed and then incubated with a fluorescein-conjugated cell-impermeable substrate (acceptor) in internalization buffer. After temperature equilibration, pre-warmed ligands were added, and time-resolved fluorescence resonance energy transfer (TR-FRET) between the donor and acceptor was measured kinetically every 3 minutes for 60 minutes at 37°C. A decrease in TR-FRET signal indicates receptor internalization. [2]
Receptor Internalization Assay (On-cell Western): HEK293 cells stably expressing HA-epitope tagged and EGFP-fused GIPR or GLP-1R were plated. After serum starvation, cells were treated with ligands at 37°C for specified times (60 min for GIPR, 30 min for GLP-1R). Cells were then fixed, blocked, and incubated with an anti-HA antibody conjugated to a fluorescent dye (DyLight800). Fluorescence signal from the cell surface was quantified using a plate scanner. Loss of surface fluorescence indicates receptor internalization. [2]
Confocal Imaging for Receptor Trafficking: HEK293 cells expressing HA- and EGFP-dual-tagged receptors were plated on imaging plates. After ligand treatment and fixation, nuclei were stained with Hoechst. Cells were imaged using a spinning-disk confocal microscope with appropriate channels for GFP (receptor localization) and Hoechst (nuclei). [2]
Ex Vivo Pancreatic Islet Perfusion: Islets were isolated from wild-type (WT) mice and mice with β-cell specific deletion of β-arrestin1 (Arrb1βcell-/-). For perfusion, 75 handpicked islets were loaded into a chamber and perfused with buffer containing low glucose (2.7 mM) for equilibration. Subsequently, the perfusate was switched to buffer containing stimulatory glucose (16 mM) alone or in combination with peptides (GLP-1, GIP, or Tirzepatide). Perfusate was collected every minute, and insulin concentration was measured by AlphaLISA. In some experiments, the GLP-1R antagonist exendin-4(9-39) or a GIPR antagonist antibody was included. [2]

High fat diet and streptozotocin injection-induced diabetic rats were injected intraperitoneally with Tirzepatide (1.35 mg/kg) once a week. The protective effects were assessed using the Morris water maze test, immunofluorescence, and Western blot analysis. Golgi staining was adopted for quantified dendritic spines.[4]
Male Sprague Dawley rats weighing between 180 and 200 g (aged 7–8 weeks) were raised in Specific Pathogen Free (SPF) conditions with a light/dark cycle of 12 h/12 h and temperature–humidity (22°C ± 1°C, 50% ± 10%) controlled. All procedures were approved by the Animal Care and Use Committee of Hubei University of Science and Technology, Xianning, China (IACUC Number: 2021-03-003). Animal care and handling were performed according to the Declaration of management of laboratory animals regarding the care and use of laboratory animals. After 2 weeks adaptation with normal diet, a total of 32 rats were fed with HF diet (67.5% standard laboratory rat chow, 20% sugar, 10% lard, 2% cholesterol and 0.5% bile salts), while 24 rats were raised by standard chow. According to our previous study, 35 mg/kg STZ was injected by intraperitoneal injection in the rats of HF diet group, whereas normal group were injected with citrate buffer only. After 2 weeks feeding, 31 rats with a fasting blood glucose levels reaching 11.0 mmol/L were randomly divided into two experimental groups as follows: diabetes mellitus group (DM), DM + Tirzepatide group (Tirzepatide, 1.35 mg/kg, once a week). At the same time, 24 rats of standard chow group were randomly divided into control group (Con) and Con + Tirzepatide group (Tirzepatide, 1.35 mg/kg, once a week). All drugs were prepared preserving more than 1 year under given conditions avoiding degradation. Oral glucose tolerance test (OGTT) was performed on the 13th week. Behavioral test was conducted before the sacrificed week. Fasting blood glucose and body weight were measured weekly until the sacrificed week. In the 15th week, all rats were sacrificed and collected samples which were executed follow-up experiments. A timeline of experimental procedure is presented in Figure 1A.[4]

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db/db mice: Weekly SC injections of Tirzepatide (1, 3, 10 nmol/kg) or vehicle for 4 weeks. HbA1c and body weight monitored weekly. [3]
Diabetic rat cognitive study: Streptozotocin-induced diabetic rats received daily SC Tirzepatide (20 nmol/kg) or saline for 8 weeks. Morris water maze tests performed at weeks 6-8; brains harvested for cytokine/AKT analysis. [4]

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This study does not describe original in vivo animal efficacy or pharmacokinetic experiments with Tirzepatide. The animal work described involves the generation and use of Arrb1βcell-/- mice for ex vivo islet perfusion experiments to investigate mechanism. Mice (Arrb1fl/fl crossed with MIP-CreERT mice) were treated with tamoxifen at 8 weeks of age (4 mg daily, orally, for 5 days) to induce β-cell specific recombination. Islets were isolated from these mice at 15-18 weeks of age for perfusion studies. [2]

Absorption, Distribution and Excretion
In the dose range of 1–5 mg, the Cmax of tezapate is 108–397 ng/mL. The mean absolute bioavailability of tezapate after subcutaneous injection is 80%. The Tmax after subcutaneous injection is 8–72 hours. Steady-state plasma concentrations are reached after weekly subcutaneous injection for 4 weeks. Because tezapate delays gastric emptying, it may affect the absorption of concurrently administered oral medications. US prescribing information recommends caution when using tezapate concomitantly with other oral medications. Tezopate is primarily excreted in urine and feces, mostly as metabolites. No altered parent drug was detected in urine or feces. The mean steady-state volume of distribution after subcutaneous administration is 9.5 L. In patients with type 2 diabetes, the mean apparent steady-state volume of distribution after subcutaneous injection of tezapate is approximately 10.3 L.
The apparent population mean clearance of tirzepatide is 0.061 L/h. The mean steady-state apparent clearance of tirzepatide is 0.056 L/h.

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Metabolism/Metabolites
Tirzepatide is metabolized by proteolysis of the peptide backbone, β-oxidation of the C20 fatty acid moiety, and amide hydrolysis.

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Biological Half-Life
The half-life is approximately five days. In humans, the half-life of subcutaneously injected tirzepatide is approximately 5 days. Steady-state plasma concentrations are reached after 4 weeks of weekly administration. [1]
The plasma clearance in patients with type 2 diabetes is 0.061 L/h. [3]

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Tirzepatide contains a C20 unsaturated diacid acyl chain that mediates its binding to albumin, thereby prolonging its half-life and enabling weekly administration in humans. [2]
Using a modified Schild regression analysis based on cell cAMP assays, which shifts the potency to the right in the presence of human serum albumin (HSA), the dissociation constant (K_A) of the interaction between tezopatide and HSA was calculated to be 1.86 μM. [2]
Based on the pharmacokinetic models of Phase I and Phase II data, the predicted steady-state total plasma concentrations (C_ss) for the clinically effective dose were: 80 nM for a 5 mg dose, 161 nM for a 10 mg dose, and 241 nM for a 15 mg dose (model predicted 15 mg dose). Using the albumin binding constant K_A and the assumed plasma albumin concentration of 640 μM, the corresponding predicted steady-state free (unbound) drug concentrations were calculated to be: 4.7 nM for a 5 mg dose, 7.0 nM for a 10 mg dose, and 10.4 nM for a 15 mg dose. [2]
At these doses, the predicted receptor occupancy (pRO) of GLP-1R, calculated using the free drug concentration and the EC₅₀ value (EC₅₀ = 533 nM) determined by low-density GLP-1R cAMP in the presence of albumin, was estimated to be 3% (5 mg), 7% (10 mg), and 10% (15 mg). For GIPR, using the corresponding albumin-biased EC₅₀ value (26.4 nM), the pRO was estimated to be 19% (5 mg), 32% (10 mg), and 41% (15 mg), reflecting its unbalanced design favoring GIPR binding. [2]

Hepatotoxicity
In pre-registration clinical trials, the incidence of serum transaminase elevations exceeding 3 times the upper limit of normal (ULN) was less than 1% in patients treated with tezapatide, with similar rates in the placebo and control groups. In studies involving over 5000 patients, no serious liver dysfunction or clinically significant liver injury caused by tezapatide has been reported. However, tezapatide was slightly associated with a higher incidence of acute gallbladder disease (gallstones, biliary cholangitis, and cholecystectomy), with an incidence of 0.6% in the treatment group and none in the placebo group. Gallbladder disease is mentioned in the warnings section of the tezapatide product label. Probability Score: E (Unlikely to cause clinically significant liver injury). Pregnancy and Lactation Effects ◉ Overview of Use During Lactation There is currently no information on the clinical use of tezapatide during lactation. Because tezoparatide is a large peptide molecule with a molecular weight of 4814 Da, its content in breast milk may be very low, and it is unlikely to be absorbed because it may be partially destroyed in the infant's gastrointestinal tract. Until more data are available, breastfeeding women should use tezoparatide with caution, especially when breastfeeding newborns or premature infants.
◉ Effects on breastfed infants
No published information found as of the revision date.
◉ Effects on lactation and breast milk
No published information found as of the revision date.
◈ What is tezoparatide?
Tezoparatide is a medication used to improve glycemic control in adults with type 2 diabetes. It is available as an injection (administered by injection). The injection is marketed under the brand name Mounjaro®. Tezoparatide is also used for the treatment of obesity by injection. Tezoparatide for weight management is marketed under the brand name Zepbound®. Weight loss is not recommended during pregnancy. If you are using Zepbound®, consult your healthcare provider before changing your medication. Your healthcare provider can discuss with you the benefits of treating your condition and the risks of not treating it during pregnancy. Obesity and high blood sugar can make pregnancy more difficult and increase the risk of miscarriage, birth defects, or other pregnancy complications. Information on diabetes (https://mothertobaby.org/fact-sheets/type-1-and-type-2-diabetes/) and obesity (https://mothertobaby.org/fact-sheets/obesity-pregnancy/) is available on the MotherToBaby website. The product label for telapatide states that using this medication may alter how oral contraceptives (pills used to prevent pregnancy) are absorbed in the body. This can increase the risk of pregnancy even if oral contraceptives are taken correctly and consistently. The product label recommends that people taking oral contraceptives switch to a non-oral contraceptive or add a barrier method of contraception (such as condoms) for 4 weeks after starting the medication and for 4 weeks after each dose increase. If you are taking this medication, discuss non-oral contraceptive methods and all options for preventing pregnancy with your healthcare provider.
◈ I am taking tinzopratide, but I want to stop taking it before I get pregnant. How long will this drug stay in my body?
The time it takes for everyone to metabolize (break down) the drug is different. For healthy adults, it takes an average of up to 30 days for most of the tinzopratide to be eliminated from the body.
◈ I am taking tinzopratide. Will it make it harder for me to get pregnant?
It is currently unclear whether tinzopratide makes it harder to get pregnant.
◈ Does taking tinzopratide increase the risk of miscarriage?
Miscarriage is common and can occur in any pregnancy for many reasons. There are currently no human studies to confirm that tinzopratide increases the risk of miscarriage.
◈ Does taking tinzopratide increase the risk of birth defects?
There is a 3-5% risk of birth defects in each pregnancy, known as background risk. There are currently no studies to confirm that tinzopratide increases the risk of birth defects in human fetuses. Animal studies have found that tinzopratide increases the risk of certain birth defects. However, it is unclear whether these birth defects are caused by the drug itself or by other factors in the studies (such as weight loss). Poorly controlled gestational diabetes during pregnancy may increase the risk of birth defects in the fetus. Controlling diabetes during pregnancy is crucial, and blood sugar levels should be kept within the target range throughout the pregnancy.
◈ Does taking tinzopratide during pregnancy increase the risk of other pregnancy-related problems?
Currently, no human studies have confirmed that tinzopratide increases the risk of pregnancy-related problems such as preterm birth (delivery before 37 weeks of gestation) or low birth weight (birth weight less than 5 pounds 8 ounces [2500 grams]). Animal studies have reported a decrease in offspring weight after exposure to tinzopratide during pregnancy. It is unclear whether this is due to the drug itself, maternal weight loss, or other factors. Poorly controlled gestational diabetes increases the risk of pregnancy complications.
◈ Will taking tinzopratide during pregnancy affect the child's future behavior or learning abilities?
Currently, no studies have assessed whether tinzopratide increases the risk of behavioral or learning problems in children.
◈ Breastfeeding while taking tinzopratide:
Currently, there is no information regarding the relationship between tinzopratide and breast milk. Because tezopratide has a large molecule size, it is not expected to enter breast milk in large quantities. Furthermore, the drug is likely to break down in the infant's gastrointestinal tract and be poorly absorbed. Please consult your healthcare provider with any questions regarding breastfeeding.
◈ Will tezopratide affect fertility or increase the risk of birth defects if the man takes it?
Currently, no human studies have evaluated whether tezopratide affects male fertility (the ability to impregnate a partner) or increases the risk of birth defects (above background risk). One animal study reported no change in male fertility. Generally, exposure to tezopratide by the father or sperm donor is unlikely to increase the risk of pregnancy. For more information, please refer to MotherToBaby's "Father Exposure" information sheet at https://mothertobaby.org/fact-sheets/paternal-exposures-pregnancy/.

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Protein Binding
Tezopratide binds to plasma albumin in 99% of its volume.

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Common adverse reactions: - Nausea (12-24%), diarrhea (12-18%) and vomiting (6-10%) (mild to moderate) occurred at doses of 5-15 mg.[1]
No drug-related serious adverse events or deaths occurred in the Phase II trial.[1]
No behavioral abnormalities or organ toxicity were observed at doses of 20 nmol/kg/day in diabetic rats.[4]

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Specific in vitro or in vivo toxicity data (e.g., LD₅₀, organ toxicity) of tezetin were not provided in this study. The study noted that dose escalation of selective GLP-1R agonists may be limited by gastrointestinal side effects such as nausea and vomiting, and GIPR activation has not yet been found to be associated with similar adverse events. Based on this characteristic, tezetin was designed with an unbalanced approach to maximize the GIPR effect while minimizing GLP-1R-related tolerability issues.[2]

[1]. Efficacy and safety of LY3298176, a novel dual GIP and GLP-1 receptor agonist, in patients with type 2 diabetes: a randomised, placebo-controlled and active comparator-controlled phase 2 trial. Lancet. 2018 Nov 17;392(10160):2180-2193.
[2]. Tirzepatide is an imbalanced and biased dual GIP and GLP-1 receptor agonist. JCI Insight. 2020 Sep 3; 5(17): e140532.
[3]. LY3298176, a novel dual GIP and GLP-1 receptor agonist for the treatment of type 2 diabetes mellitus: From discovery to clinical proof of concept. Mol Metab. 2018 Dec:18:3-14.
[4]. Tirzepatide ameliorates spatial learning and memory impairment through modulation of aberrant insulin resistance and inflammation response in diabetic rats. Front Pharmacol. 2023 Aug 28;14:1146960.

Pharmacodynamics
Tilatide is a synthetic peptide with hypoglycemic effects. It exerts its glucose-dependent effects by stimulating insulin secretion in both the first and second phases and reducing glucagon levels. Tilatide has also been shown to delay gastric emptying, reduce fasting and postprandial blood glucose concentrations, decrease food intake, and reduce weight in patients with type 2 diabetes. Tilatide can improve insulin sensitivity. Because the peptide is coupled to the C20 fatty acid moiety at the 20th lysine residue via a hydrophilic linker, the drug is highly bound to albumin in plasma, thus prolonging its half-life. Background: LY3298176 is a novel dual glucose-dependent insulinotropic peptide (GIP) and glucagon-like peptide-1 (GLP-1) receptor agonist currently under development for the treatment of type 2 diabetes. This study aimed to investigate the efficacy and safety of LY3298176 in patients with poorly controlled type 2 diabetes, compared with placebo or dulaglutide, which selectively stimulates the GLP-1 receptor. Methods: In this double-blind, randomized, phase II study, patients with type 2 diabetes were randomized in a 1:1:1:1:1 ratio to receive once-weekly subcutaneous injections of LY3298176 (1 mg, 5 mg, 10 mg, or 15 mg), dulaglutide (1.5 mg), or placebo for 26 weeks. Grouping was stratified based on baseline glycated hemoglobin A1c (HbA1c), metformin use, and body mass index (BMI). Eligible participants (aged 18–75 years) with type 2 diabetes for at least 6 months (HbA1c 7.0–10.5%, inclusive 7.0% and 10.5%), whose glycemic control was not adequately achieved by diet and exercise or stable metformin therapy alone, and whose body mass index (BMI) was 23–50 kg/m2. The primary efficacy endpoint was the change in HbA1c from baseline to 26 weeks in the modified intention-to-treat (mITT) population (all patients who had received at least one treatment with the study drug and had at least one post-baseline measurement of any outcome measure). Secondary endpoints were measured in the mITT treatment dataset and included changes in HbA1c from baseline to week 12; changes in mean weight, fasting blood glucose, waist circumference, total cholesterol, LDL cholesterol, HDL cholesterol, and triglycerides; and changes in the proportion of patients achieving HbA1c targets (≤6.5% and <7.0%) from baseline to week 12 and week 26. The proportion of patients achieving at least 5% and 10% weight loss from baseline to week 26 were also included. This study was registered at ClinicalTrials.gov under accession number NCT03131687. Results: Eligibility for enrollment was assessed for 555 participants between May 24, 2017, and March 28, 2018, of whom 318 were randomized to one of the six treatment groups. Due to two untreated participants, the modified intention-to-treat and safety analysis populations comprised a total of 316 participants. 258 (81.7%) participants completed 26 weeks of treatment, and 283 (89.6%) completed the study. At baseline, the mean age was 57 years (standard deviation 9), the BMI was 32.6 kg/m² (5.9), the duration of diabetes diagnosis was 9 years (6), the HbA1c was 8.1% (1.0), 53% of patients were male and 47% were female. At week 26, the effect of LY3298176 on HbA1c changes was dose-dependent and did not reach a plateau. Compared with placebo, the mean changes in HbA1c from baseline after treatment with LY3298176 were: -1.06% in the 1 mg group, -1.73% in the 5 mg group, -1.89% in the 10 mg group, and -1.94% in the 15 mg group (compared to -0.06% in the placebo group) (posterior mean differences [80% confidence interval] compared with placebo: -1.00% [-1.22 to -0.79] in the 1 mg group, -1.67% [-1.88 to -1.46] in the 5 mg group, -1.83% [-2.04 to -1.61] in the 10 mg group, and -1.89% [-2.11 to -1.67] in the 15 mg group). Compared with dulaglutide (-1.21%), the posterior mean differences (80% confidence set) in HbA1c changes from baseline to 26 weeks with LY3298176 dose were: 0.15% (-0.08 to 0.38) in the 1 mg dose group, -0.52% (-0.72 to -0.31) in the 5 mg dose group, -0.67% (-0.89 to -0.46) in the 10 mg dose group, and -0.73% (-0.95 to -0.52) in the 15 mg dose group. At week 26, among patients treated with LY3298176, 33% to 90% achieved the target HbA1c level below 7.0% (52% in the dulaglutide group and 12% in the placebo group), and 15% to 82% achieved the target HbA1c level of at least 6.5% (39% in the dulaglutide group and 2% in the placebo group). Fasting blood glucose levels in the LY3298176 group ranged from -0.4 mmol/L to -3.4 mmol/L (0.9 mmol/L in the placebo group and -1.2 mmol/L in the dulaglutide group). Mean weight loss in the LY3298176 group ranged from -0.9 kg to -11.3 kg (0.4 kg in the placebo group and -2.7 kg in the dulaglutide group). At week 26, among patients treated with LY3298176, 14% to 71% achieved at least 5% of their weight loss target (22% in the dulaglutide group, 0% in the placebo group), and 6% to 39% achieved at least 10% of their weight loss target (9% in the dulaglutide group, 0% in the placebo group). Waist circumference changes in the LY3298176 group ranged from -2.1 cm to -10.2 cm (-1.3 cm in the placebo group, -2.5 cm in the dulaglutide group). Total cholesterol changes in the LY3298176 group ranged from 0.2 mmol/L to -0.3 mmol/L (0.3 mmol/L in the placebo group, -0.2 mmol/L in the dulaglutide group). There were no significant differences in HDL or LDL cholesterol changes between the LY3298176 and placebo groups. Triglyceride concentrations in the LY3298176 group ranged from 0 mmol/L to -0.8 mmol/L (0.3 mmol/L in the placebo group and -0.3 mmol/L in the dulaglutide group). The 12-week and 26-week results for all secondary endpoints were similar. Of the 316 subjects across the six treatment groups, 13 (4%) experienced 23 serious adverse events. Gastrointestinal events (nausea, diarrhea, and vomiting) were the most common adverse events occurring during treatment. The incidence of gastrointestinal adverse events was dose-related (23.1% in the 1 mg LY3298176 group, 32.7% in the 5 mg LY3298176 group, 51.0% in the 10 mg LY3298176 group, 66.0% in the 15 mg LY3298176 group, 42.6% in the dulaglutide group, and 9.8% in the placebo group); most adverse events were mild to moderate and transient. Decreased appetite was the second most common adverse event (3.8% in the 1 mg LY3298176 group, 20.0% in the 5 mg LY3298176 group, 25.5% in the 10 mg LY3298176 group, 18.9% in the 15 mg LY3298176 group, 5.6% in the dulaglutide group, and 2.0% in the placebo group). No serious hypoglycemic events were reported. One patient in the placebo group died of stage IV lung adenocarcinoma, unrelated to the study treatment. Conclusion: The dual GIP and GLP-1 receptor agonist LY3298176 showed more significant efficacy than dulaglutide in glycemic control and weight loss, with acceptable safety and tolerability. Combined stimulation of GIP and GLP-1 receptors may provide a new treatment option for type 2 diabetes. [1]

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Teratriide (LY3298176) is a dual GIP and GLP-1 receptor agonist being developed for the treatment of type 2 diabetes (T2DM), obesity, and non-alcoholic steatohepatitis. Early clinical trials of T2DM have shown that terazetide is superior to selective GLP-1 receptor agonists in improving clinical efficacy. Therefore, we hypothesize that the combined potency and signal transduction properties of terazetide give it unique pharmacological characteristics that can effectively improve a wide range of metabolic controls. This paper establishes a method for calculating the occupancy of each receptor at the clinically effective dose of this drug. Analysis results showed that tezepatide binds to the GIP receptor more strongly than to the GLP-1 receptor, confirming the imbalance of its mechanism of action. Pharmacological signal transduction studies showed that tezepatide mimics the action of natural GIP on the GIP receptor, but exhibits bias on the GLP-1 receptor, tending to promote cAMP production rather than β-arrestin recruitment, and its ability to drive GLP-1 receptor internalization is also weaker than that of GLP-1. Primary islet experiments showed that β-arrestin1 limited the insulin response of GLP-1 rather than GIP or tezepatide, suggesting that the biased agonist effect of tezepatide enhances insulin secretion. The GIP receptor imbalance, coupled with the unique signal transduction properties of the GLP-1 receptor, may jointly explain the good efficacy of the study drug. [2]

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Objective: To develop a novel dual GIP and GLP-1 receptor agonist, LY3298176, to determine whether the metabolic effects of GIP can enhance the established clinical benefits of selective GLP-1 receptor agonists in type 2 diabetes mellitus (T2DM). Methods: LY3298176 is a fatty acid-modified peptide with dual GIP and GLP-1 receptor agonist activity, designed for once-weekly subcutaneous injection. In vitro, LY3298176 was characterized using cell lines expressing recombinant or endogenous incretin receptors for signal transduction and functional analysis. In vivo, LY3298176 was characterized by mouse body weight, food intake, insulin secretion, and blood glucose profile. A phase I randomized, placebo-controlled, double-blind study was conducted in three parts: first, a single-dose escalation (SAD; dose 0.25–8 mg) study and a 4-week multiple-dose escalation (MAD; dose 0.5–10 mg) study in healthy subjects (HS); followed by a 4-week phase Ib multiple-dose proof-of-concept (POC; dose 0.5–15 mg) study in patients with type 2 diabetes mellitus (T2DM) (ClinicalTrials.gov registration number: NCT02759107). Doses higher than 5 mg were obtained by titration, with dulaglutide (DU) used as a positive control. The primary objective of this study was to investigate the safety and tolerability of LY3298176. Results: LY3298176 activated the GIP and GLP-1 receptor signaling pathways in vitro and, in mice, demonstrated glucose-dependent insulin secretion and improved glucose tolerance by acting on GIP and GLP-1 receptors. Long-term administration of LY3298176 significantly reduced body weight and food intake in mice; these effects were significantly stronger than those of GLP-1 receptor agonists. A total of 142 subjects received at least one dose of LY3298176, dulaglutide, or placebo. Pharmacokinetic studies of LY3298176 were conducted over a wide dose range (0.25–15 mg), and the results supported a once-weekly dosing regimen. In a phase 1b trial in diabetic patients, LY3298176 at doses of 10 mg and 15 mg significantly reduced fasting blood glucose compared to placebo (least square mean [LSM] difference [95% CI]: -49.12 mg/dL [-78.14, -20.12] and -43.15 mg/dL [-73.06, -13.21], respectively). In patients with MAD HS, the LY3298176 1.5 mg, 4.5 mg, and 10 mg dose groups showed significantly greater weight loss than the placebo group (least square mean difference [95% CI]: -1.75 kg [-3.38, -0.12], -5.09 kg [-6.72, -3.46], and -4.61 kg [-6.21, -3.01], respectively). The 10 mg and 15 mg dose groups also showed significant efficacy in patients with type 2 diabetes mellitus (least square mean difference [95% CI]: -2.62 kg [-3.79, -1.45] and -2.07 kg [-3.25, -0.88], respectively). The most common adverse reactions to LY3298176 were gastrointestinal reactions (vomiting, nausea, decreased appetite, diarrhea, and abdominal distension), which occurred in both patients with hepatitis B (HS) and type 2 diabetes mellitus. All adverse reactions were dose-dependent and mild to moderate in severity. Conclusion: Based on these results, the pharmacological properties of LY3298176 have been translated from preclinical studies to clinical studies. LY3298176 has the potential to provide clinically meaningful improvements in glycemic control and weight. These data support further clinical evaluation of LY3298176 for the treatment of type 2 diabetes mellitus and potential obesity. [3]

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Tilatin is a single peptide constructed by integrating GLP-1 activity into the GIP sequence. It is described as an “unbalanced and biased” dual agonist. Unbalanced refers to its higher affinity and potency for GIPR than for GLP-1R. Biased refers to its signal transduction properties on GLP-1R, which, unlike the natural ligand GLP-1, preferentially activates the cAMP pathway rather than recruiting β-arrestin. [2]
Biased agonist effect: Compared to GLP-1, the internalization effect of GLP-1R is weakened. In vitro perfusion experiments of Arrb1βcell-/- mouse islets showed that the absence of β-arrestin1 enhanced the insulin secretion response of GLP-1, but had no effect on GIP or tirzepatide, suggesting that the biased signaling of tirzepatide on GLP-1R may enhance its insulin secretion-promoting effect by avoiding β-arrestin-mediated restriction. [2] The good clinical efficacy of tirzepatide is thought to be due to the following: 1) complete and potent GLP-1R agonism; 2) unbalanced binding that favors GLP-1R, thus allowing for higher doses and potentially better tolerability; 3) biased signaling on GLP-1R may lead to enhanced insulin secretion. [2]
Tirzepatide is under development for the treatment of type 2 diabetes (T2DM), obesity, and non-alcoholic steatohepatitis (NASH). [2]

C225H348N48O68

4813.45147800446

4810.52

C, 56.14; H, 7.29; N, 13.97; O, 22.60

2023788-19-2

Tirzepatide hydrochloride; Tirzepatide TFA;13C,15N Tirzepatide;Tirzepatide TFA (LY3298176 TFA);Tirzepatide hydrochloride (LY3298176 hydrochloride); 2933217-72-0 (sodium salt); 2931515-08-9 (acetate)

168009818

Tyr-{Aib}-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Ile-{Aib}-Leu-Asp-Lys-Ile-Ala-Gln-{C20 diacid-gamma-Glu-(AEEA)2-Lys}-Ala-Phe-Val-Gln-Trp-Leu-Ile-Ala-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-NH2

Y-{Aib}-EGTFTSDYSI-{Aib}-LDKIAQ-{C20 diacid-gamma-Glu-(AEEA)2-Lys}-AFVQWLIAGGPSSGAPPPS-NH2; or YXEGTFTSDY SIXLDKIAQK AFVQWLIAGG PSSGAPPPS

White to off-white solid powder

95.0~105.0%

-6.8

58

70

163

341

11700

0

CC[C@H](C)[C@@H](C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CCCCNC(=O)COCCOCCNC(=O)COCCOCCNC(=O)CC[C@H](C(=O)O)NC(=O)CCCCCCCCCCCCCCCCCCC(=O)O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CC2=CNC3=CC=CC=C32)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(=O)NCC(=O)NCC(=O)N4CCC[C@H]4C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](C)C(=O)N5CCC[C@H]5C(=O)N6CCC[C@H]6C(=O)N7CCC[C@H]7C(=O)N[C@@H](CO)C(=O)N)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CC(C)C)NC(=O)C(C)(C)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@H](CO)NC(=O)[C@H](CC8=CC=C(C=C8)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CC9=CC=CC=C9)NC(=O)[C@H]([C@@H](C)O)NC(=O)CNC(=O)[C@H](CCC(=O)O)NC(=O)C(C)(C)NC(=O)[C@H](CC1=CC=C(C=C1)O)N.CC(=O)O

BTSOGEDATSQOAF-SMAAHMJQSA-N

InChI=1S/C225H348N48O68/c1-23-126(10)183(264-198(311)146(64-50-52-88-226)246-202(315)157(109-180(297)298)252-199(312)152(103-124(6)7)261-223(337)225(21,22)269-217(330)185(128(12)25-3)266-209(322)163(120-278)257-200(313)153(107-138-74-78-141(282)79-75-138)250-203(316)158(110-181(299)300)253-207(320)162(119-277)259-216(329)187(134(18)280)267-206(319)155(106-136-60-44-41-45-61-136)254-215(328)186(133(17)279)262-174(289)114-237-193(306)147(83-87-179(295)296)260-222(336)224(19,20)268-192(305)143(227)104-137-72-76-140(281)77-73-137)214(327)242-131(15)190(303)244-148(80-84-168(228)283)196(309)245-145(65-51-53-89-231-175(290)121-340-100-99-339-97-91-233-176(291)122-341-101-98-338-96-90-232-170(285)86-82-150(221(334)335)243-171(286)70-46-38-36-34-32-30-28-26-27-29-31-33-35-37-39-47-71-178(293)294)195(308)240-130(14)191(304)248-154(105-135-58-42-40-43-59-135)205(318)263-182(125(8)9)212(325)247-149(81-85-169(229)284)197(310)251-156(108-139-111-234-144-63-49-48-62-142(139)144)201(314)249-151(102-123(4)5)204(317)265-184(127(11)24-2)213(326)241-129(13)189(302)236-112-172(287)235-115-177(292)270-92-54-66-164(270)210(323)258-161(118-276)208(321)256-160(117-275)194(307)238-113-173(288)239-132(16)218(331)272-94-56-68-166(272)220(333)273-95-57-69-167(273)219(332)271-93-55-67-165(271)211(324)255-159(116-274)188(230)301/h40-45,48-49,58-63,72-79,111,123-134,143,145-167,182-187,234,274-282H,23-39,46-47,50-57,64-71,80-110,112-122,226-227H2,1-22H3,(H2,228,283)(H2,229,284)(H2,230,301)(H,231,290)(H,232,285)(H,233,291)(H,235,287)(H,236,302)(H,237,306)(H,238,307)(H,239,288)(H,240,308)(H,241,326)(H,242,327)(H,243,286)(H,244,303)(H,245,309)(H,246,315)(H,247,325)(H,248,304)(H,249,314)(H,250,316)(H,251,310)(H,252,312)(H,253,320)(H,254,328)(H,255,324)(H,256,321)(H,257,313)(H,258,323)(H,259,329)(H,260,336)(H,261,337)(H,262,289)(H,263,318)(H,264,311)(H,265,317)(H,266,322)(H,267,319)(H,268,305)(H,269,330)(H,293,294)(H,295,296)(H,297,298)(H,299,300)(H,334,335)/t126-,127-,128-,129-,130-,131-,132-,133+,134+,143-,145-,146-,147-,148-,149-,150+,151-,152-,153-,154-,155-,156-,157-,158-,159-,160-,161-,162-,163-,164-,165-,166-,167-,182-,183-,184-,185-,186-,187-/m0/s1

20-[[(1R)-4-[2-[2-[2-[2-[2-[2-[[(5S)-5-[[(2S)-5-amino-2-[[(2S)-2-[[(2S,3S)-2-[[(2S)-6-amino-2-[[(2S)-2-[[(2S)-2-[[2-[[(2S,3S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S,3R)-2-[[(2S)-2-[[(2S,3R)-2-[[2-[[(2S)-2-[[2-[[(2S)-2-amino-3-(4-hydroxyphenyl)propanoyl]amino]-2-methylpropanoyl]amino]-4-carboxybutanoyl]amino]acetyl]amino]-3-hydroxybutanoyl]amino]-3-phenylpropanoyl]amino]-3-hydroxybutanoyl]amino]-3-hydroxypropanoyl]amino]-3-carboxypropanoyl]amino]-3-(4-hydroxyphenyl)propanoyl]amino]-3-hydroxypropanoyl]amino]-3-methylpentanoyl]amino]-2-methylpropanoyl]amino]-4-methylpentanoyl]amino]-3-carboxypropanoyl]amino]hexanoyl]amino]-3-methylpentanoyl]amino]propanoyl]amino]-5-oxopentanoyl]amino]-6-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-5-amino-1-[[(2S)-1-[[(2S)-1-[[(2S,3S)-1-[[(2S)-1-[[2-[[2-[(2S)-2-[[(2S)-1-[[(2S)-1-[[2-[[(2S)-1-[(2S)-2-[(2S)-2-[(2S)-2-[[(2S)-1-amino-3-hydroxy-1-oxopropan-2-yl]carbamoyl]pyrrolidine-1-carbonyl]pyrrolidine-1-carbonyl]pyrrolidin-1-yl]-1-oxopropan-2-yl]amino]-2-oxoethyl]amino]-3-hydroxy-1-oxopropan-2-yl]amino]-3-hydroxy-1-oxopropan-2-yl]carbamoyl]pyrrolidin-1-yl]-2-oxoethyl]amino]-2-oxoethyl]amino]-1-oxopropan-2-yl]amino]-3-methyl-1-oxopentan-2-yl]amino]-4-methyl-1-oxopentan-2-yl]amino]-3-(1H-indol-3-yl)-1-oxopropan-2-yl]amino]-1,5-dioxopentan-2-yl]amino]-3-methyl-1-oxobutan-2-yl]amino]-1-oxo-3-phenylpropan-2-yl]amino]-1-oxopropan-2-yl]amino]-6-oxohexyl]amino]-2-oxoethoxy]ethoxy]ethylamino]-2-oxoethoxy]ethoxy]ethylamino]-1-carboxy-4-oxobutyl]amino]-20-oxoicosanoic acid

LY-3298176; LY 3298176; tirzepatide; LY3298176; BG 121; BG121; BG-121

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: 1) Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. 2) This product is not stable in solution, please use freshly prepared working solution for optimal results.

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

DMSO: ~50 mg/mL (~10.4 mM)
Ethanol : 8~9 mg/mL
Water : Insoluble

Note: Please refer to the "Guidelines for Dissolving Peptides" section in the 4th page of the "Instructions for use" file (upper-right section of this webpage) for how to dissolve peptides.

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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.)