GLP-1 receptor
Glucagon-like peptide 1 (GLP-1) receptor agonist. [1]
Glucagon-like peptide-1 receptor (GLP-1R). Semaglutide is a long-acting GLP-1 analogue that activates GLP-1 receptors. [1, 2]
The target of semaglutide is the glucagon-like peptide-1 receptor (GLP-1R), a G protein-coupled receptor widely distributed across pancreatic β-cells, α-cells, heart, vascular smooth muscle cells, central nervous system (hypothalamus and brainstem), kidneys, gastrointestinal tract, and other tissues. By specifically binding to GLP-1R, semaglutide activates downstream signaling pathways, promoting insulin secretion in a glucose-dependent manner, suppressing glucagon release, delaying gastric emptying, and inhibiting appetite centers, thereby achieving dual effects of glucose lowering and weight reduction.

Semaglutide is derivatized at lysine 26 and differs from human GLP-1 in two amino acid substitutions (Aib⁸, Arg³⁴). Semaglutide has an affinity for GLP-1R of 0.38±0.06 nM[1]. With a 94% sequence homology to human GLP-1, semaglutide is an analogue of GLP-1[3]. In vitro studies demonstrate that semaglutide exhibits high affinity and potent agonistic activity at the human GLP-1 receptor. Radioligand displacement assays show an IC50 value of 0.38 nM (pIC50 = 9.42) for semaglutide. In a luciferase reporter assay using BHK cells expressing the human GLP-1 receptor, semaglutide achieved an EC50 value of 0.01 nM (pEC50 = 11.21), indicating significant GLP-1 receptor activation at nanomolar to picomolar concentrations. This high potency positions semaglutide as one of the most active GLP-1 analogues available.

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Semaglutide targets the GLP-1 receptor (GLP-1R), a G protein-coupled receptor primarily expressed in pancreatic β-cells, as well as in the brain, gastrointestinal tract, and other tissues . Activation of GLP-1R stimulates adenylate cyclase, increasing intracellular cyclic AMP (cAMP), which promotes glucose-dependent insulin secretion and inhibits glucagon release .
Binding affinity data :
- IC50 (membrane radioligand displacement assay, ¹²⁵I-GLP-1 as tracer): 0.38 nM (pIC50 = 9.42)

cAMP accumulation assay (CHO cells expressing human GLP-1R) :
- EC50 (0.1% BSA): 12 pM (pEC50 = 11.21)
- Maximal effect: 100.7% relative to GLP-1

Effect of human serum albumin on potency :
- EC50 in 4.4% HSA (physiological albumin concentration): 2,630 pM
- This ~220-fold shift in EC50 reflects semaglutide's high albumin binding affinity, which is responsible for its extended half-life

Activity in human pancreatic β-cells (EndoC-βH1) :
- EC50 (0.1% BSA): 9,748 pM (approximately 12,000-fold higher than GLP-1)
- Maximal effect: 128.3% (superior to GLP-1)

Semaglutide has an MRT of 63.6 hours after s.c. dosing to mini-pigs and a plasma half-life of 46 hours in mini-pigs after intravenous administration[1]. Motor impairments caused by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) are ameliorated by semaglutide. Semaglutide also protects dopaminergic neurons in the substantia nigra and striatum by rescuing the decrease in tyrosine hydroxylase (TH) levels, reducing lipid peroxidation, alleviating inflammation, inhibiting the apoptosis pathway, and increasing the expression of autophagy-related proteins. Furthermore, semaglutide, the long-acting GLP-1 analogue, outperforms NN-2211 in the majority of parameters[2].

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In a randomized, double-blind, placebo-controlled cardiovascular outcomes trial (SUSTAIN-6), 3297 patients with type 2 diabetes at high cardiovascular risk received once-weekly subcutaneous semaglutide (0.5 mg or 1.0 mg) or placebo for 104 weeks. The primary composite outcome (cardiovascular death, nonfatal myocardial infarction, or nonfatal stroke) occurred in 6.6% of patients in the semaglutide group vs. 8.9% in the placebo group (hazard ratio, 0.74; 95% CI, 0.58 to 0.95; P<0.001 for noninferiority; P=0.02 for superiority). [1]
Nonfatal stroke occurred in 1.6% vs. 2.7% (hazard ratio, 0.61; 95% CI, 0.38 to 0.99; P=0.04). Nonfatal myocardial infarction occurred in 2.9% vs. 3.9% (hazard ratio, 0.74; 95% CI, 0.51 to 1.08; P=0.12). Cardiovascular death occurred in 2.7% vs. 2.8% (hazard ratio, 0.98; 95% CI, 0.65 to 1.48; P=0.92). [1]
At week 104, glycated hemoglobin levels decreased by 1.1% (0.5 mg dose) and 1.4% (1.0 mg dose) from a baseline of 8.7% in the semaglutide groups, compared to a decrease of 0.4% in the placebo groups (P<0.001 for both doses). [1]
Body weight decreased by 3.6 kg (0.5 mg dose) and 4.9 kg (1.0 mg dose) from a baseline of 92.1 kg, compared to decreases of 0.7 kg and 0.5 kg in the placebo groups (P<0.001 for both doses). [1]
New or worsening nephropathy occurred in 3.8% of the semaglutide group vs. 6.1% of the placebo group (hazard ratio, 0.64; 95% CI, 0.46 to 0.88; P=0.005). [1]
Diabetic retinopathy complications occurred in 3.0% of the semaglutide group vs. 1.8% of the placebo group (hazard ratio, 1.76; 95% CI, 1.11 to 2.78; P=0.02). [1]

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Cardiovascular Outcomes in Type 2 Diabetes (SUSTAIN-6 Trial): In a randomized, double-blind, placebo-controlled trial involving 3297 patients with type 2 diabetes at high cardiovascular risk, once-weekly semaglutide (0.5 mg or 1.0 mg) for 104 weeks significantly reduced the primary composite outcome of cardiovascular death, nonfatal myocardial infarction, or nonfatal stroke (6.6% in semaglutide group vs. 8.9% in placebo group; hazard ratio, 0.74; 95% CI, 0.58 to 0.95; P<0.001 for noninferiority; P=0.02 for superiority). Nonfatal stroke occurred in 1.6% of semaglutide-treated patients vs. 2.7% in placebo group (hazard ratio, 0.61; 95% CI, 0.38 to 0.99; P=0.04). Nonfatal myocardial infarction occurred in 2.9% vs. 3.9% (hazard ratio, 0.74; 95% CI, 0.51 to 1.08; P=0.12). Cardiovascular death was similar between groups (2.7% vs. 2.8%; hazard ratio, 0.98; 95% CI, 0.65 to 1.48; P=0.92). New or worsening nephropathy was lower in the semaglutide group (3.8% vs. 6.1%; hazard ratio, 0.64; 95% CI, 0.46 to 0.88; P=0.005). [1]
Glycemic Control: At week 104, semaglutide reduced mean glycated hemoglobin from 8.7% at baseline to 7.6% (0.5 mg) and 7.3% (1.0 mg), representing changes of -1.1% and -1.4%, respectively, compared to -0.4% in placebo groups. [1]
Body Weight: Semaglutide reduced mean body weight by -3.6 kg (0.5 mg) and -4.9 kg (1.0 mg) at week 104, compared to -0.7 kg and -0.5 kg in placebo groups. [1]
Blood Pressure: Semaglutide (1.0 mg) reduced mean systolic blood pressure by 5.4 mm Hg at week 104, compared to 2.8 mm Hg in placebo group (estimated treatment difference -2.6 mm Hg; P<0.001). [1]
Heart Rate: Semaglutide increased mean pulse rate by 2.4 bpm (1.0 mg) at week 104, compared to a decrease of 0.1 bpm in placebo group (estimated treatment difference +2.5 bpm; P<0.001). [1]
Neuroprotective Effects in MPTP Mouse Model of Parkinson's Disease: In male C57BL/6 mice, once-daily intraperitoneal injection of semaglutide (25 nmol/kg) for 7 days significantly improved MPTP-induced motor impairments. Semaglutide increased the distance traveled in open-field test, increased time on rotarod, and improved stride length abnormalities in footprint test compared to MPTP-only group (P<0.001 for all). Semaglutide restored tyrosine hydroxylase (TH)-positive dopaminergic neuron numbers in substantia nigra (to 88.56% of control vs. 59.18% in MPTP group) and TH expression in striatum (to 72.43% of control vs. 57.00% in MPTP group). Semaglutide also reduced astrogliosis (GFAP) and microgliosis (IBA-1), decreased lipid peroxidation (4-HNE), reduced apoptosis (increased Bcl-2, decreased Bax, reduced Bax/Bcl-2 ratio), and increased autophagy-related proteins (beclin1, Atg7, LC3, P62) compared to MPTP-only group. Semaglutide was more effective than liraglutide at the same dose in most parameters measured. [2]

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In vivo, semaglutide exerts multiple pharmacological effects through GLP-1 receptor activation. In glycemic control, it promotes insulin secretion in a glucose-dependent manner while suppressing glucagon release, effectively lowering both fasting and postprandial blood glucose. In weight management, semaglutide reduces appetite and increases satiety by acting on the satiety center in the hypothalamus, while also delaying gastric emptying, thereby significantly reducing caloric intake. Clinical studies have demonstrated that once-weekly subcutaneous administration of 2.4 mg semaglutide achieves approximately 15% body weight reduction. Additionally, semaglutide exhibits cardiovascular protective effects, reducing the risk of major adverse cardiovascular events in patients with type 2 diabetes.

HEK293‐SNAP‐GLP‐1R cells were labelled in suspension with SNAP‐Lumi4‐Tb (40 nM, Cisbio, Codelet, France) for 1 hour at room temperature in complete medium. After washing and resuspension in hanks' balanced salt solution containing 0.1% bovine serum albumin and metabolic inhibitors (20 mmol/L 2‐deoxygucose and 10 mmol/L NaN3) to prevent GLP‐1R internalization, binding experiments were performed by time‐resolved förster resonance energy transfer (FRET) using exendin (9‐39) with fluorescein isothiocyanate (FITC) installed at position K12 as previously described. [4]

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DPP-4 resistance :
- Native GLP-1 is rapidly degraded by dipeptidyl peptidase-4 (DPP-4) within 2-3 minutes
- Semaglutide is fully stable against DPP-4 due to the substitution of alanine at position 8 with α-aminoisobutyric acid (Aib), a non-natural amino acid that prevents enzyme recognition and cleavage

In vitro gastrointestinal enzyme stability :
Semaglutide half-life (T1/2) in various enzyme systems:
- Chymotrypsin: <5.0 minutes
- Trypsin: 9.3 minutes
- Pancreatin: 7.9 minutes
- Pepsin: 60.1 minutes
- Elastase: 2.4 minutes

Plasma stability :
- Semaglutide demonstrates significantly enhanced stability in plasma compared to native GLP-1, attributable to both DPP-4 resistance and albumin binding that protects the peptide from proteolytic degradation

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1. Membrane Preparation: Extract cell membranes from cells overexpressing the human GLP-1 receptor, and purify membrane proteins using homogenization and differential centrifugation. 2. Radioligand Binding Assay: Incubate membrane proteins with radiolabeled tracer (e.g., ¹²⁵I-GLP-1) and various concentrations of semaglutide in binding buffer at room temperature or 37°C for 60-120 minutes. 3. Separation and Counting: Terminate the reaction by rapid filtration using glass fiber filters, wash filters 3-5 times with wash buffer to remove unbound ligands, dry, and count radioactivity using a gamma counter. 4. Data Analysis: Calculate IC50 values by fitting competition curves and convert to Ki values using the Cheng-Prusoff equation to evaluate the binding affinity of semaglutide to the GLP-1 receptor.

Semaglutide activates the GLP-1 receptor in pancreatic beta cells leading to glucose-dependent insulin release. It also decreases glucagon secretion, slows gastric emptying, and promotes satiety.
Hepa 1-6 hepatocyte model (NAFLD study) :
- Semaglutide treatment (in vivo and in vitro) significantly reduced ABHD6 expression and increased miR-5120 expression in high glucose + free fatty acid-stimulated Hepa 1-6 cells
- The GLP-1R was confirmed to be widely expressed in Hepa 1-6 cells and mouse liver tissue
- Semaglutide regulated the miR-5120/ABHD6 axis through GLP-1R activation, alleviating hepatic steatosis

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1. Cell Culture: Culture BHK cells (or CHO cells) expressing the human GLP-1 receptor in DMEM medium containing 10% fetal bovine serum at 37°C in 5% CO₂ until 80-90% confluence. 2. Cell Seeding: After trypsinization, seed cells at appropriate density (e.g., 2×10⁴/well) into 96-well white plates and culture overnight. 3. Compound Treatment: Remove medium, add stimulation buffer containing various concentrations of semaglutide (ranging from 0.001 nM to 100 nM, 10-fold serial dilutions) and 0.1% BSA, and incubate at 37°C for 30-60 minutes. 4. Reporter Assay: Add luciferase detection reagent, incubate at room temperature protected from light for 5-10 minutes, and measure chemiluminescence signal using a microplate reader. 5. Data Analysis: Generate concentration-response curves and calculate EC50 values.

Mice: Male C57BL/6 mice 10 weeks old (20-25 g) are used throughout the study. Six groups of mice are randomly assigned (n = 12 per group). The treatments were as follows: (i) saline alone was given to the control group; (ii) NN-2211 group received saline and NN-2211 (25 nmol/kg ip. once daily for 7 days); (iii) Semaglutide group received saline and Semaglutide (25 nmol/kg ip. once daily for 7 days); (iv) MPTP group received MPTP alone (once daily 20 mg/kg ip. for 7 days); (v) MPTP (once daily 20 mg/kg ip. for 7 days) was immediately followed by NN-2211 treated group (25 nmol/kg ip. once daily for 7 days). (vi) MPTP (20 mg/kg i.p. once daily for 7 days), which was immediately followed by the group treated with semaglutide (25 nmol/kg i.p. once daily for 7 days). Measure behavioral changes, neuronal damage, inflammatory markers, and other biomarkers at the conclusion of drug treatments[2].

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Parkinson's disease was modeled in C57BL/6 mice using 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Mice received MPTP hydrochloride (30 mg/kg, ip.) once daily for 5 consecutive days. Control groups received saline. [2]
Drug treatment groups received either semaglutide or liraglutide at a dose of 25 mg/kg (ip.), once daily for 7 days, starting from the first day of MPTP injection. Control groups for the drugs received saline injections. [2]
Motor functions were assessed using behavioral tests: 1) Open-field test to evaluate spontaneous locomotor and exploratory activity. 2) Rotarod test to assess bradykinesia and balance. 3) Footprint test to analyze gait and posture. [2]
Following behavioral tests, mice were perfused, and brains were collected. Brain sections containing the substantia nigra and striatum were prepared for immunohistochemical analysis. Tyrosine hydroxylase (TH) staining was performed to quantify dopaminergic neurons and terminals. Immunohistochemistry was also conducted for GFAP, IBA-1, 4-HNE, Bcl-2, and Bax. Stained sections were visualized under a light microscope, and quantitative analysis (cell counts or optical density) was performed using image analysis software. [2]
For Western blot analysis, substantia nigra tissue was homogenized in lysis buffer. Protein concentration was determined, and samples were separated by SDS-PAGE, transferred to PVDF membranes, and probed with primary antibodies against Bcl-2, Bax, Beclin1, Atg7, LC3, P62, and β-Actin (loading control). After incubation with HRP-conjugated secondary antibodies, protein bands were visualized using enhanced chemiluminescence and quantified. [2]

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1. Animal Models: Use diet-induced obese (DIO) mice or high-fat diet-fed rodent models, weighing approximately 30-40 g. 2. Dosing Regimen: Administer semaglutide via subcutaneous injection at doses typically ranging from 3-100 nmol/kg, once or twice weekly; administer vehicle control (e.g., PBS) to the control group. 3. Body Weight and Food Intake Monitoring: Measure body weight 2-3 times daily or weekly, and record daily food and water intake. 4. Blood Glucose Measurement: Measure blood glucose via tail-tip blood collection at various time points post-dose (e.g., 0, 2, 4, 8, 12, 24 hours), or perform intraperitoneal glucose tolerance testing (IPGTT) to assess glucose regulation. 5. Metabolic Parameter Assessment: Evaluate energy expenditure, respiratory quotient, and other parameters using metabolic cage systems, and analyze body composition (fat mass/lean mass) by NMR. 6. Tissue Collection: Euthanize animals at the experimental endpoint and collect tissues such as pancreas, hypothalamus, and liver for further analysis.

Absorption, Distribution and Excretion
In a clinical trial, semaglutide's Cmax was 10.9 nmol/L, AUC was 3123.4 nmol·h/L, and Tmax was 56 h, reached within 1–3 days. Absolute bioavailability was 89%. Steady-state concentrations of oral tablets were reached within 4–5 weeks. The mean steady-state concentration of semaglutide refers to the mean steady-state concentration after administration of a dose from 0.5 mg to 1 mg, ranging from 16 nmol/L to 30 nmol/L. The drug is primarily cleared by the kidneys and can be excreted in urine and feces. The primary route of clearance is urine, equivalent to 53% of the ingested radiolabeled dose, followed by feces, approximately 18.6%. A small amount (3.2%) is excreted via exhalation. Hepatic impairment does not appear to affect the clearance of the drug, and no dose adjustment is required for patients with impaired hepatic function. The volume of distribution of semaglutide is 8 L to 9.4 L. It crosses the rat placenta.
According to a clinical study, the clearance rate of semaglutide is 0.039 L/h. The FDA label indicates that the clearance rate of semaglutide in patients with type 2 diabetes is approximately 0.05 L/h.
Metabolism/Metabolites

Semaglutide is cleaved at the peptide backbone, followed by β-oxidation of the fatty acid chain. Naturally occurring GLP-1 is rapidly metabolized by dipeptidyl peptidase-4 (DPP-4) and other enzymes widely present in human tissues. Chemical modifications make semaglutide less susceptible to degradation by gastrointestinal DPP-4 enzymes. It is metabolized slowly and extensively, with approximately 83% of the administered dose remaining in plasma as the parent drug. Neuroendopeptidase (NEP) is another enzyme that metabolizes this drug. DPP-4 inactivates semaglutide by cleaving the N-terminal fragment, while NEP hydrolyzes peptide bonds. Six different semaglutide metabolites have been identified in human plasma. The major metabolite P3 accounts for approximately 7.7% of the ingested dose.

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Biological Half-Life
One of the main characteristics of semaglutide is its long half-life of 168 hours. This long half-life is attributed to its binding to albumin. This reduces renal clearance and protects semaglutide from metabolic degradation.

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Semaglutide is a GLP-1 analog with a half-life of approximately one week, thus it can be administered subcutaneously once weekly. [1]

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Semaglutide is a long-acting GLP-1 analogue with an extended half-life of approximately 1 week, allowing once-weekly dosing. It is a modification of liraglutide that is protease-resistant by changing the amino acid at position 8 (to aminoisobutyric acid). [1, 2]
In the MPTP mouse study, semaglutide was administered intraperitoneally at 25 nmol/kg once daily for 7 days. [2]

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Semaglutide exhibits favorable long-acting pharmacokinetic properties. Following subcutaneous administration, its half-life is approximately 165 hours (approximately 1 week), supporting once-weekly dosing. This extended half-life results from two key structural modifications: substitution of the 8th amino acid with 2-aminoisobutyric acid to resist DPP-4 degradation, and conjugation of the 26th lysine to a C18 fatty diacid chain via a linker, enhancing albumin binding and reducing renal clearance. The oral formulation (Rybelsus®) achieves gastric absorption through co-formulation with the absorption enhancer SNAC, with an oral bioavailability of approximately 1%. Semaglutide is primarily metabolized via peptide hydrolysis without involvement of specific cytochrome P450 pathways, and metabolites are excreted in urine and feces.

Hepatotoxicity
In large clinical trials, the incidence of elevated serum enzymes was not higher in the semaglutide treatment group compared to placebo or control drugs, and no clinically significant cases of liver injury were reported. In fact, semaglutide and other GLP-1 analogues often improve serum transaminase levels (and hepatic steatosis), making it a potential treatment for non-alcoholic fatty liver disease. Since its market launch, no case reports of hepatotoxicity caused by semaglutide have been published, and liver injury is not listed as an adverse event in the product information. Therefore, liver injury caused by semaglutide, even if it occurs, is certainly very rare. Probability score: E (unlikely to be the cause of clinically significant liver injury). Pregnancy and Lactation Effects ◉ Overview of Use During Lactation There is currently no information regarding the clinical use of semaglutide during lactation. Because semaglutide is a peptide molecule with a molecular weight of 4113 Da and a protein binding rate of over 99%, its content in breast milk is likely to be very low. Furthermore, the oral absorption rate of semaglutide is only 0.4% to 1%, therefore it is unlikely to have adverse effects on breastfed infants.
◉ Effects on breastfed infants
No published information found as of the revision date.
◉ Effects on breastfeeding and breast milk
No published information found as of the revision date.
◈ What is semaglutide?
Semaglutide is a medication used to improve glycemic control in adults with type 2 diabetes. It is available in both injectable (administered by injection) and tablet (oral) formulations. The injectable formulation is marketed as Ozempic®. The tablet formulation is marketed as Rybelsus®. Semaglutide is also available as an injectable treatment for obesity. Semaglutide for weight management is marketed as Wegovy®. Weight loss is not recommended during pregnancy. Please discuss with your healthcare provider how to use Wegovy® and which treatment regimen is best for you. Always consult with your healthcare provider before changing your medication regimen. 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/poorly controlled blood sugar can make conception more difficult and increase the risk of miscarriage, birth defects, or other pregnancy complications. The MotherToBaby website provides information on diabetes (https://mothertobaby.org/fact-sheets/type-1-and-type-2-diabetes/) and obesity (https://mothertobaby.org/fact-sheets/obesity-pregnancy/).
◈ I am taking semaglutide, but I want to stop taking it before I get pregnant. How long will this medication stay in my body?
Everyone's drug metabolism rate is different. In healthy adults, it takes an average of up to 6 weeks for most of semaglutide to be eliminated from the body. The product information for Ozempic®, Wegovy®, and Rybelsus® recommends that women planning to become pregnant stop taking this medication 2 months before conception.
◈ I am taking semaglutide. Will it affect my ability to get pregnant?
It is currently unclear whether semaglutide affects pregnancy.
◈ Does taking semaglutide increase the risk of miscarriage?
Miscarriage is common and can occur in any pregnancy for a variety of reasons. No human studies have been conducted to determine whether semaglutide increases the risk of miscarriage. Animal studies have reported that taking semaglutide increases the risk of miscarriage. It is unclear whether this finding is due to the drug itself or to weight loss. Because there are many causes of miscarriage, it is difficult to determine whether the drug, illness, or other factors caused the miscarriage.
◈ Does taking semaglutide increase the risk of birth defects?
There is a 3-5% risk of birth defects in each pregnancy, known as background risk. No studies have confirmed that semaglutide increases the risk of birth defects in human fetuses. There has been one case report of pregnancy while taking semaglutide. The patient continued taking semaglutide for the first 3-4 weeks of pregnancy and had a successful delivery without any reported birth defects. In animal studies conducted by the manufacturer, an increased risk of certain birth defects has been observed. However, this only occurred when the mother was given a dose of semaglutide that was toxic to the animal. Furthermore, it is currently unclear whether the reported birth defects were caused by the medication or other factors in the study, such as weight loss. Since high blood sugar/poor blood sugar control increases the risk of birth defects and other pregnancy complications, it is crucial to weigh the benefits of using semaglutide against the risks of not treating it. Consult your healthcare provider to learn the best way to treat your condition during pregnancy.
◈ Does taking semaglutide during pregnancy increase the risk of other pregnancy-related problems?
There are currently no human studies to determine whether semaglutide 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 that when parent animals were exposed to doses higher than those used in humans, their offspring were smaller than normal. It is currently unclear whether this is due to the medication, weight loss, or because the study animals were healthy enough to remain healthy without taking semaglutide.
◈ Will taking semaglutide during pregnancy affect a child's future behavior or learning abilities?
There are currently no studies exploring whether semaglutide causes behavioral or learning problems in children.
◈ Breastfeeding while taking semaglutide:
There is currently no information regarding the relationship between semaglutide and breast milk. Based on one animal study, a small amount of semaglutide is expected to enter breast milk. Your healthcare provider can discuss the use of semaglutide with you and the best treatment option for you. Please be sure to consult your healthcare provider about all breastfeeding-related questions. The Rybelsus® product label advises that women who are breastfeeding should not use the tablet form of this medication if they are nursing an infant. This is because there is a theoretical concern that taking this medication in tablet form may result in higher drug concentrations in the nursing infant. However, the benefits of using semaglutide may outweigh the potential risks. Your healthcare provider can discuss the use of semaglutide in other dosage forms (tablets or injections) and which treatment option is best for you.
◈ Will taking semaglutide affect fertility or increase the risk of birth defects in men?
No human studies have been conducted to determine whether semaglutide affects male fertility (the ability to impregnate a partner) or increases the risk of birth defects (above background risk). An animal study using the same dose of semaglutide as in humans showed no changes in male fertility. Generally, exposure to semaglutide by the father or sperm donor is unlikely to increase the risk of pregnancy. For more information, see the “Paternal Exposure” information sheet on the MotherToBaby website at https://mothertobaby.org/fact-sheets/paternal-exposures-pregnancy/.

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Protein Binding
Semaglutide binds to plasma albumin with high affinity, which ensures high stability of the drug. Its binding rate to albumin is over 99%.

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The incidence of gastrointestinal disorders (nausea, vomiting, diarrhea) was higher in the semaglutide group compared to the placebo group. Most were mild or moderate and occurred within the first 30 weeks of treatment. Discontinuation of treatment due to adverse events (mainly gastrointestinal adverse events) was more common in the semaglutide group. [1]
The incidence of serious adverse events was lower in the semaglutide group. [1] The incidence of diabetic retinopathy complications was significantly higher in the semaglutide group than in the placebo group (hazard ratio, 1.76; 95% CI, 1.11 to 2.78; P = 0.02). [1] Nine patients in the semaglutide group developed acute pancreatitis, compared to 12 in the placebo group; all events were mild. [1] The incidence of serious or symptomatic hypoglycemia was similar in both the semaglutide and placebo groups (approximately 22% in both groups). [1] The incidence of malignancies was similar in both the semaglutide and placebo groups. One patient in the semaglutide group (1.0 mg dose) developed pancreatic cancer, compared to four patients in the placebo group. No medullary thyroid carcinoma was diagnosed. [1]

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Adverse Events in SUSTAIN-6: Gastrointestinal disorders were more frequent in the semaglutide group than placebo (50.7-52.3% vs. 35.2-35.7%), with nausea (17.3-21.9% vs. 7.5-8.1%), vomiting (10.5-14.8% vs. 4.1-5.2%), and diarrhea (17.9-18.4% vs. 10.5-11.9%) being the most common. Treatment discontinuation due to adverse events (mainly gastrointestinal) was more frequent in the semaglutide group (11.5-14.5% vs. 5.7-7.6%). Serious adverse events were lower in the semaglutide group (33.6-35.0% vs. 36.1-39.9%). [1]
Diabetic Retinopathy Complications: Semaglutide was associated with a higher risk of retinopathy complications (vitreous hemorrhage, blindness, or need for intravitreal agent or photocoagulation) compared to placebo (3.0% vs. 1.8%; hazard ratio, 1.76; 95% CI, 1.11 to 2.78; P=0.02). Of 79 patients with retinopathy complications, 66 (83.5%) had preexisting retinopathy at baseline. [1]
Pancreatitis and Pancreatic Cancer: Acute pancreatitis occurred in 9 patients in the semaglutide group vs. 12 in placebo group. Pancreatic cancer occurred in 1 patient receiving semaglutide (1.0 mg) vs. 4 patients receiving placebo. No medullary thyroid carcinomas were confirmed in either group. [1]
In MPTP Mouse Model: Semaglutide was well-tolerated at the tested dose (25 nmol/kg/day i.p. for 7 days) with no reported adverse effects on general health or behavior. [2]

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The toxicological profile of semaglutide has been extensively evaluated in both preclinical and clinical studies. The most common adverse reactions are gastrointestinal, including nausea (approximately 44%), vomiting (approximately 25%), diarrhea, and constipation; these are mostly mild to moderate in severity, occur primarily during the initial treatment period, and lead to treatment discontinuation in approximately 4.3% of patients. Thyroid C-cell tumor risk is an important safety concern for semaglutide, with observations of C-cell hyperplasia and medullary thyroid carcinoma in animal studies; consequently, the drug is contraindicated in patients with a personal or family history of medullary thyroid carcinoma. Other notable adverse events include acute pancreatitis, gallbladder disease, worsening of diabetic retinopathy, and increased heart rate. In non-clinical safety assessments, semaglutide showed no significant signs of toxicity at doses up to 192 mg/kg, indicating a favorable safety margin. The drug is contraindicated during pregnancy and within 2 months prior to planned pregnancy due to potential risks to the fetus.

[1]. Semaglutide and Cardiovascular Outcomes in Patients with Type 2 Diabetes. N Engl J Med. 2016 Nov 10;375(19):1834-1844.

[2]. Neuroprotective effects of the novel GLP-1 long acting analogue semaglutide in the MPTP Parkinson's disease mouse model. Neuropeptides. 2018 Oct;71:70-80.

[3]. Semaglutide: First Global Approval. Drugs. 2018 Feb;78(2):275-284.

Pharmacodynamics
Semaglutide lowers glycated hemoglobin (HbA1c), systolic blood pressure, and body weight. After 12 weeks of treatment, semaglutide lowers fasting and postprandial blood glucose by increasing insulin secretion and decreasing glucagon secretion (glucagon is typically associated with elevated blood glucose). Semaglutide also lowers fasting triglycerides and very low-density lipoprotein cholesterol (VLDL-C), thus having a beneficial effect on cardiovascular health. Studies have shown that semaglutide can induce medullary thyroid carcinoma in rodents. Although its clinical significance in humans is unclear, the U.S. Food and Drug Administration (FDA) recommends that patients with a personal or family history of medullary thyroid carcinoma should not use this drug. Semaglutide also carries the risk of pancreatitis and dehydration. Patients must maintain adequate hydration while taking semaglutide, and if abdominal pain radiating to the back occurs, immediate medical attention is recommended. Because this drug delays gastric emptying, monitoring the efficacy or adverse reactions of other oral medications is crucial.

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Semaglutide was in the development stage for type 2 diabetes at the time of this trial. The SUSTAIN-6 trial was a premarket cardiovascular outcome trial designed to assess its non-inferiority compared to placebo. [1]
The trial showed that semaglutide significantly reduced the risk of major adverse cardiovascular events (cardiovascular death, non-fatal myocardial infarction, or non-fatal stroke) in patients with type 2 diabetes at high cardiovascular risk, confirming its non-inferiority and demonstrating superiority. [1]
This beneficial cardiovascular effect may be associated with a reduction in glycated hemoglobin, body weight, and systolic blood pressure, as well as a potential improvement in the progression of atherosclerosis. [1]
An important safety concern found was an increased risk of diabetic retinopathy complications. [1]

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Semaglutide is a long-acting GLP-1 analogue approved for the treatment of type 2 diabetes (brand name Ozempic). It is a modification of liraglutide with an amino acid substitution at position 8 (to aminoisobutyric acid) that confers protease resistance, resulting in a half-life of approximately 1 week and allowing once-weekly subcutaneous administration. [1, 2]
In the SUSTAIN-6 cardiovascular outcomes trial, semaglutide met its primary noninferiority endpoint and demonstrated superiority over placebo for reducing major adverse cardiovascular events in patients with type 2 diabetes at high cardiovascular risk. The number needed to treat to prevent one primary outcome event over 24 months was 45. [1]
In the MPTP mouse model of Parkinson's disease, semaglutide demonstrated neuroprotective effects, improving motor function, preserving dopaminergic neurons, reducing inflammation and oxidative stress, inhibiting apoptosis, and enhancing autophagy. Semaglutide was more effective than liraglutide at the same dose in most parameters, suggesting potential utility for treating Parkinson's disease. [2]

C187H291N45O59

4113.5776

4111.12

C, 54.60; H, 7.13; N, 15.32; O, 22.95

910463-68-2

1997361-85-9 (Semaglutide acetate); 910463-68-2 (Semaglutide free base); 2924330-56-1 (sodium)

56843331

H-His-Aib-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys-Glu-Phe-Ile-Ala-Trp-Leu-Val-Arg-Gly-Arg-Gly-OH

HXEGTFTSDV SSYLEGQAAK EFIAWLVRGR G

White to off-white solid powder

-5.8

57

63

151

291

9590

30

CC[C@H](C)[C@@H](C(=O)N[C@@H](C)C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCNC(=N)N)C(=O)NCC(=O)N[C@@H](CCCNC(=N)N)C(=O)NCC(=O)O)NC(=O)[C@H](CC3=CC=CC=C3)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CCCCNC(=O)COCCOCCNC(=O)COCCOCCNC(=O)CC[C@H](C(=O)O)NC(=O)CCCCCCCCCCCCCCCCC(=O)O)NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@H](CCC(=O)N)NC(=O)CNC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC4=CC=C(C=C4)O)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@H](C(C)C)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CC5=CC=CC=C5)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](CC6=CN=CN6)N

DLSWIYLPEUIQAV-CCUURXOWSA-N

InChI=1S/C187H291N45O59/c1-18-105(10)154(180(282)208-108(13)159(261)216-133(86-114-89-200-119-50-40-39-49-117(114)119)170(272)218-129(82-102(4)5)171(273)228-152(103(6)7)178(280)215-121(53-44-72-199-186(192)193)162(264)201-91-141(242)209-120(52-43-71-198-185(190)191)161(263)204-94-151(257)258)230-172(274)131(83-111-45-33-31-34-46-111)219-167(269)126(64-69-149(253)254)214-166(268)122(51-41-42-70-195-144(245)98-290-79-78-289-76-74-197-145(246)99-291-80-77-288-75-73-196-139(240)66-61-127(183(285)286)211-140(241)54-37-29-27-25-23-21-19-20-22-24-26-28-30-38-55-146(247)248)212-158(260)107(12)206-157(259)106(11)207-165(267)125(60-65-138(189)239)210-142(243)92-202-163(265)123(62-67-147(249)250)213-168(270)128(81-101(2)3)217-169(271)130(85-113-56-58-116(238)59-57-113)220-175(277)135(95-233)223-177(279)137(97-235)224-179(281)153(104(8)9)229-174(276)134(88-150(255)256)221-176(278)136(96-234)225-182(284)156(110(15)237)231-173(275)132(84-112-47-35-32-36-48-112)222-181(283)155(109(14)236)227-143(244)93-203-164(266)124(63-68-148(251)252)226-184(287)187(16,17)232-160(262)118(188)87-115-90-194-100-205-115/h31-36,39-40,45-50,56-59,89-90,100-110,118,120-137,152-156,200,233-238H,18-30,37-38,41-44,51-55,60-88,91-99,188H2,1-17H3,(H2,189,239)(H,194,205)(H,195,245)(H,196,240)(H,197,246)(H,201,264)(H,202,265)(H,203,266)(H,204,263)(H,206,259)(H,207,267)(H,208,282)(H,209,242)(H,210,243)(H,211,241)(H,212,260)(H,213,270)(H,214,268)(H,215,280)(H,216,261)(H,217,271)(H,218,272)(H,219,269)(H,220,277)(H,221,278)(H,222,283)(H,223,279)(H,224,281)(H,225,284)(H,226,287)(H,227,244)(H,228,273)(H,229,276)(H,230,274)(H,231,275)(H,232,262)(H,247,248)(H,249,250)(H,251,252)(H,253,254)(H,255,256)(H,257,258)(H,285,286)(H4,190,191,198)(H4,192,193,199)/t105-,106-,107-,108-,109+,110+,118-,120-,121-,122-,123-,124-,125-,126-,127+,128-,129-,130-,131-,132-,133-,134-,135-,136-,137-,152-,153-,154-,155-,156-/m0/s1

18-[[(1R)-4-[2-[2-[2-[2-[2-[2-[[(5S)-5-[[(2S)-2-[[(2S)-2-[[(2S)-5-amino-2-[[2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-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-(1H-imidazol-5-yl)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-methylbutanoyl]amino]-3-hydroxypropanoyl]amino]-3-hydroxypropanoyl]amino]-3-(4-hydroxyphenyl)propanoyl]amino]-4-methylpentanoyl]amino]-4-carboxybutanoyl]amino]acetyl]amino]-5-oxopentanoyl]amino]propanoyl]amino]propanoyl]amino]-6-[[(2S)-1-[[(2S)-1-[[(2S,3S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-5-carbamimidamido-1-[[2-[[(2S)-5-carbamimidamido-1-(carboxymethylamino)-1-oxopentan-2-yl]amino]-2-oxoethyl]amino]-1-oxopentan-2-yl]amino]-3-methyl-1-oxobutan-2-yl]amino]-4-methyl-1-oxopentan-2-yl]amino]-3-(1H-indol-3-yl)-1-oxopropan-2-yl]amino]-1-oxopropan-2-yl]amino]-3-methyl-1-oxopentan-2-yl]amino]-1-oxo-3-phenylpropan-2-yl]amino]-4-carboxy-1-oxobutan-2-yl]amino]-6-oxohexyl]amino]-2-oxoethoxy]ethoxy]ethylamino]-2-oxoethoxy]ethoxy]ethylamino]-1-carboxy-4-oxobutyl]amino]-18-oxooctadecanoic acid

NN 9535; NN9535; NN-9535; Ozempic; NNC 0113-0217; NNC-0113-0217; NNC0113-0217; Semaglutide; Ozempic; Rybelsus; NN9535; UNII-53AXN4NNHX; Wegovy; NN 9535;

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, avoid exposure to moisture.

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

H2O: ~50 mg/mL (~12.2 mM)
DMSO: ~5 mg/mL (~1.2 mM)

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.)