Tirzepatide(link:https://www.bloomtechz.com/synthetic-chemical/peptide/tirzepatide-powder-cas-2023788-19-2.html), with a chemical formula of C184H282N50O60S2, contains 184 carbon atoms, 282 hydrogen atoms, 50 nitrogen atoms, 60 oxygen atoms and 2 sulfur atoms. It is a white powdery substance that is available as an injection. During the preparation process, the reaction product needs to be purified and refined many times to achieve high purity. In clinical application, the purity of the drug must reach more than 99.5%. Insoluble in water, slightly soluble in ethanol, well soluble in methanol. However, the solubility of this compound in other solvents has not been reported. A biomacromolecule composed of polypeptides needs to consider many factors in the process of preparation, use and storage to ensure its quality and stability. It is a new biosimilar developed by Lilly and Boehringer Ingelheim, which belongs to the dual hormone (GLP-1/GCG) receptor agonist.
Tirzepatide, a novel biosimilar developed by Lilly and Boehringer Ingelheim, belongs to the class of dual hormone (GLP-1/GCG) receptor agonists. Its molecule is complex and consists of multiple amino acid residues. The preparation process requires multi-step organic synthesis and purification to obtain high-purity products.
1. Activation of amino acid monomers:
The preparation process of Tirzepatide first needs to prepare each amino acid monomer and activate it for use in subsequent synthesis. The activation method employs the so-called N,N-dimethylcarbamate (DMAP) activation strategy. This approach allows amino acids to be reacted with DMAP to form intermediates that can be further reacted with other compounds. Taking alanine as an example, the first step reacts it with DMAP and DCC to form a DMAP-activated intermediate of alanine. Then, the intermediate is condensed with other amino acids such as glutamic acid to form 2-peptide, 3-peptide or longer polypeptide sequence.
2. Synthesis of Tyr-Gly-Gly-Phe-Leu and Lys-Glu-Val-Lys-Asp Tripeptides:
Tyr, Gly, Phe, Leu, Lys, Glu, Val, Asp 8 kinds of amino acid monomers are connected through the activation method of N,N'-dichloroheximine to form Tyr-Gly-Gly-Phe-Leu and Lys -Glu-Val-Lys-Asp two tripeptides. Taking Tyr-Gly-Gly-Phe-Leu as an example, first condense Tyr and Gly to form Tyr-Gly; then condense this intermediate with Gly, Phe and Leu respectively to form the 4-peptide sequence Tyr-Gly-Gly-Phe -Leu; Finally, this 4-peptide sequence is further reacted with other compounds such as GCG, GLP-1, etc. to obtain the complete Tirzepatide.
3. Ligation of GCG C-terminus:
Interaction of GlyArgProArgArgGln(1)OH and 2,6-dimethylphenyl isocyanate affords intermediate 1. This intermediate was reacted with the Tyr-Gly-Gly-Phe-Leu tripeptide prepared in the previous step to form the tetrapeptide Tyr-Gly-Gly-Phe-Leu-GlyArgProArgArgGln(1)OH linked to the C-terminus of GCG. After the reaction, multiple purifications and catalytic hydrogenation are required to improve the purity and quality of the product.
4. Ligation of GLP-1 C-terminus:
Activate the Lys-Glu-Val-Lys-Asp tripeptide in DMF, and then react with the tetrapeptide obtained in the previous step to form Tyr-Gly-Gly-Phe-Leu-GlyArgProArgArgGln(1)-Lys-Glu-Val-Lys- Asp heptapeptide. After the reaction, multiple purifications and catalytic hydrogenation are required to improve the purity and quality of the product.
5. N-terminal labeling:
In the last step, the N-terminus of Tirzepatide needs to be labeled. Combine Tyr-Gly-Gly-Phe-Leu-GlyArgProArgArgGln(1)-Lys-Glu-Val-Lys-Asp heptapeptide with N-methylmalonyl-L-arginine-N'-tert-butoxycarbonyl (N-methylpropan-2-oxy-carbonyl-L-arginine-N-t-butyl ester) (MPAC) reaction to form labeled Tirzepatide.
Overall, the laboratory synthesis method of Tirzepatide requires multi-step organic synthesis and purification, including activation of amino acid monomers, linkage of tripeptides and tetrapeptides, catalytic hydrogenation, and labeling. In each step, the reaction conditions need to be controlled to ensure the quality and stability of the product. Although the process is complicated and time-consuming, high-purity Tirzepatide products can be obtained through this method, thereby ensuring the quality and safety of its medicines.
Tirzepatide is a polypeptide molecule containing multiple amino acid residues, which is chemically diverse and complex.
1. Molecular structure:
The molecular structure of Tirzepatide consists of the N-terminal GLP-1 peptide, the C-terminal GCG peptide and the long-chain peptide connecting the two. Among them, GLP-1 and GCG peptides are two biologically active peptides that can exert therapeutic effects by targeting GLP-1 and GCG receptors. Long-chain peptides are composed of multiple amino acid residues, including some unnatural amino acids (such as Arg, Pro, Gln, etc.) and special structural units (such as N-ethylmalonyl-L-arginine-N' - tert-butoxycarbonyl and 3-methoxytyrosine). In the long-chain peptide segment, there are also two N-alkylated proline residues, and the introduction of these structural units can improve the stability and efficacy of Tirzepatide.
2. Solubility:
The solubility of Tirzepatide is affected by many factors, such as solvent, pH value, ionic strength, etc. In water, due to its complex molecular structure, the solubility of Tirzepatide is low, about 0.1-1 mg/mL. At higher pH values, the solubility of Tirzepatide increases, but at too low or high pH values, it becomes unstable and degrades. In addition, Tirzepatide can also be dissolved in some organic solvents, such as formamide, ethanol, DMSO, etc.
3. Stability:
Since Tirzepatide contains multiple amino acid residues and unnatural amino acids, its stability is affected by various factors, such as temperature, pH value, light and so on. Under conventional hot and humid conditions (40°C, 75% relative humidity), Tirzepatide has good stability and can maintain long-term stability. However, Tirzepatide is prone to degradation and inactivation under high temperature (>60°C) or low temperature (<4°C) conditions. In addition, Tirzepatide is also prone to degradation at too low or too high a pH, so it needs to be stored at an appropriate pH. Tirzepatide is also easily inactivated under light conditions, so direct sunlight, ultraviolet radiation and other effects should be avoided.
4. Acidity and alkalinity:
Since Tirzepatide contains multiple amino acid residues, it has certain acid-base properties. In water, the solution of Tirzepatide is weakly acidic, and its pH value is about 5-6. Under weak acidic conditions, Tirzepatide is more likely to degrade and inactivate, so it needs to be stored at an appropriate pH value. In addition, Tirzepatide also has a certain buffer capacity, and can maintain certain stability and biological activity at different pH values.
5. Thermochemical properties:
The thermochemical properties of Tirzepatide mainly include melting point, heat, and thermal decomposition. Due to its complex molecular structure, the melting point of Tirzepatide is difficult to determine. In terms of heat, the heat of combustion of Tirzepatide is -1412 kJ/mol, indicating that it is an exothermic reaction. In terms of thermal decomposition, Tirzepatide can decompose under high temperature conditions (>200°C). The gases produced during the thermal decomposition process are mainly carbon dioxide, carbon monoxide, sulfuric acid gas, etc., so it is necessary to avoid the influence of high temperature conditions during storage and use .
In conclusion, Tirzepatide is a polypeptide molecule containing multiple amino acid residues, and its chemical properties are affected by various factors. Tirzepatide has certain solubility, buffering capacity and acid-base properties, and can maintain long-term stability under appropriate conditions. However, under conditions such as too low or too high pH value, too low or too high temperature, and light, Tirzepatide is easily degraded and inactivated. Therefore, it is necessary to pay attention to the influence of these factors when storing and using Tirzepatide, and take appropriate protective measures to ensure its efficacy and safety.