Tirzepatide(link:https://www.bloomtechz.com/synthetic-chemical/peptide/tirzepatide-powder-cas-2023788-19-2.html), nrog cov tshuaj formula C184H282N50O60S2, muaj 184 carbon atoms, 282 hydrogen atoms, 50 nitrogen atoms, 60 oxygen atoms thiab 2 sulfur atoms. Nws yog cov hmoov dawb dawb uas muaj nyob rau hauv kev txhaj tshuaj. Thaum lub sij hawm kev npaj, cov tshuaj tiv thaiv cov khoom yuav tsum tau purified thiab refined ntau zaus kom ua tau siab purity. Hauv kev siv tshuaj kho mob, qhov purity ntawm cov tshuaj yuav tsum ncav cuag ntau dua 99.5 feem pua. Insoluble hauv dej, me ntsis soluble hauv ethanol, zoo soluble hauv methanol. Txawm li cas los xij, qhov solubility ntawm qhov sib xyaw no hauv lwm cov kuab tshuaj tsis tau tshaj tawm. Ib qho biomacromolecule muaj li ntawm polypeptides yuav tsum xav txog ntau yam hauv cov txheej txheem ntawm kev npaj, siv thiab khaws cia kom ntseeg tau tias nws zoo thiab ruaj khov. Nws yog ib tug tshiab biosimilar tsim los ntawm Lilly thiab Boehringer Ingelheim, uas belongs rau dual hormone (GLP{12}}/GCG) receptor agonist.
Tirzepatide, ib tug tshiab biosimilar tsim los ntawm Lilly thiab Boehringer Ingelheim, belongs rau chav kawm ntawm dual hormone (GLP-1/GCG) receptor agonists. Nws cov molecule yog complex thiab muaj ntau yam amino acid residues. Cov txheej txheem kev npaj yuav tsum muaj ntau cov txheej txheem organic synthesis thiab purification kom tau txais cov khoom zoo purity.
1. Ua kom cov amino acid monomers:
Cov txheej txheem kev npaj ntawm Tirzepatide thawj zaug yuav tsum tau npaj txhua cov amino acid monomer thiab ua kom nws siv rau hauv kev sib txuas tom qab. Txoj kev ua kom ua haujlwm siv lub npe hu ua N, N-dimethylcarbamate (DMAP) ua kom lub tswv yim. Txoj hauv kev no tso cai rau cov amino acids tau hnov mob nrog DMAP los tsim cov intermediates uas tuaj yeem cuam tshuam ntxiv nrog lwm cov tebchaw. Noj alanine ua piv txwv, thawj kauj ruam reacts nws nrog DMAP thiab DCC los tsim DMAP-activated intermediate ntawm alanine. Tom qab ntawd, qhov nruab nrab yog condensed nrog lwm cov amino acids xws li glutamic acid los ua 2-peptide, 3-peptide lossis ntev dua polypeptide ib ntus.
2. Synthesis ntawm Tyr-Gly-Gly-Phe-Leu thiab Lys-Glu-Val-Lys-Asp Tripeptides:
Tyr, Gly, Phe, Leu, Lys, Glu, Val, Asp 8 hom amino acid monomers txuas nrog los ntawm txoj kev ua kom N, N'-dichloroheximine los ua Tyr-Gly-Gly-Phe-Leu thiab Lys-Glu- Val-Lys-Asp ob tripeptides. Siv Tyr-Gly-Gly-Phe-Leu ua piv txwv, thawj condense Tyr thiab Gly los ua Tyr-Gly; ces condense no intermediate nrog Gly, Phe thiab Leu feem los ua tus 4-peptide sequence Tyr-Gly-Gly-Phe -Leu; Thaum kawg, qhov no 4-peptide ib ntus yog txuas ntxiv nrog rau lwm cov tebchaw xws li GCG, GLP-1, thiab lwm yam kom tau txais Tirzepatide tiav.
3. Ligation ntawm GCG C-terminus:
Kev sib cuam tshuam ntawm GlyArgProArgArgGln(1)OH thiab 2, 6-dimethylphenyl isocyanate affords intermediate 1. Qhov nruab nrab no tau cuam tshuam nrog Tyr-Gly-Gly-Phe-Leu tripeptide npaj hauv cov kauj ruam dhau los los tsim cov tetrapeptide Tyr-Gly- Gly-Phe-Leu-GlyArgProArgArgGln(1)OH txuas rau C-terminus ntawm GCG. Tom qab cov tshuaj tiv thaiv, ntau yam purifications thiab catalytic hydrogenation yuav tsum tau txhim kho cov purity thiab zoo ntawm cov khoom.
4. Ligation ntawm GLP-1 C-terminus:
Ua kom cov Lys-Glu-Val-Lys-Asp tripeptide hauv DMF, thiab tom qab ntawd hnov mob nrog tetrapeptide tau txais hauv cov kauj ruam dhau los los ua Tyr-Gly-Gly-Phe-Leu-GlyArgProArgArgGln(1)-Lys-Glu-Val-Lys - Asp heptapeptide. Tom qab cov tshuaj tiv thaiv, ntau yam purifications thiab catalytic hydrogenation yuav tsum tau txhim kho cov purity thiab zoo ntawm cov khoom.
5. N-terminal labeling:
Hauv kauj ruam kawg, N-terminus ntawm Tirzepatide yuav tsum tau sau npe. Muab Tyr-Gly-Gly-Phe-Leu-GlyArgProArgArgGln(1)-Lys-Glu-Val-Lys-Asp heptapeptide nrog N-methylmalonyl-L-arginine-N'-tert-butoxycarbonyl (N-methylpropan{19}} oxy-carbonyl-L-arginine-Nt-butyl ester) (MPAC) cov tshuaj tiv thaiv rau daim ntawv sau npe Tirzepatide.
Zuag qhia tag nrho, lub chaw soj nstuam synthesis txoj kev ntawm Tirzepatide yuav tsum tau ntau-kauj ruam organic synthesis thiab purification, nrog rau kev ua kom cov amino acid monomers, linkage ntawm tripeptides thiab tetrapeptides, catalytic hydrogenation, thiab daim ntawv lo. Hauv txhua kauj ruam, cov kev mob tshwm sim yuav tsum tau tswj xyuas kom zoo thiab ruaj khov ntawm cov khoom. Txawm hais tias cov txheej txheem nyuaj thiab siv sijhawm ntev, cov khoom siv siab huv Tirzepatide tuaj yeem tau txais los ntawm txoj kev no, yog li ua kom cov tshuaj zoo thiab kev nyab xeeb ntawm nws cov tshuaj.
Tirzepatide yog polypeptide molecule uas muaj ntau yam amino acid residues, uas yog chemically ntau haiv neeg thiab complex.
1. Molecular qauv:
Cov qauv molecular ntawm Tirzepatide muaj N-terminal GLP-1 peptide, C-terminal GCG peptide thiab cov saw ntev peptide txuas ob. Ntawm lawv, GLP-1 thiab GCG peptides yog ob qho tshuaj lom neeg peptides uas tuaj yeem siv tshuaj kho mob los ntawm kev tsom mus rau GLP-1 thiab GCG receptors. Long-chain peptides yog tsim los ntawm ntau cov amino acid residues, suav nrog qee cov amino acids tsis zoo (xws li Arg, Pro, Gln, thiab lwm yam) thiab cov qauv tshwj xeeb (xws li N-ethylmalonyl-L-arginine-N '- tert-butoxycarbonyl thiab 3-methoxytyrosine). Hauv ntu ntev-peptide ntu, kuj tseem muaj ob N-alkylated proline residues, thiab kev qhia txog cov qauv txheej txheem no tuaj yeem txhim kho kev ruaj ntseg thiab kev ua tau zoo ntawm Tirzepatide.
2. Solubility:
Lub solubility ntawm Tirzepatide yog cuam tshuam los ntawm ntau yam, xws li hnyav, pH nqi, ionic zog, thiab lwm yam. Nyob rau hauv dej, vim nws complex molecular qauv, lub solubility ntawm Tirzepatide yog tsawg, txog 0.{{1} } mg/ml. Thaum pH siab dua, qhov solubility ntawm Tirzepatide nce, tab sis ntawm qhov qis los yog siab pH qhov tseem ceeb, nws yuav tsis ruaj khov thiab degrades. Tsis tas li ntawd, Tirzepatide kuj tuaj yeem yaj hauv qee cov kuab tshuaj organic, xws li formamide, ethanol, DMSO, thiab lwm yam.
3. Kev ruaj ntseg:
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 degree) lossis qis kub (<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 thiab alkalinity:
Txij li Tirzepatide muaj ntau yam amino acid residue, nws muaj qee yam acid-puag zog. Hauv dej, kev daws ntawm Tirzepatide yog acidic tsis muaj zog, thiab nws tus nqi pH yog li ntawm 5-6. Nyob rau hauv tsis muaj zog acidic tej yam kev mob, Tirzepatide feem ntau yuav degrade thiab inactivated, yog li nws yuav tsum tau muab cia rau ntawm ib tug tsim nyog pH nqi. Tsis tas li ntawd, Tirzepatide kuj muaj qee qhov tsis muaj peev xwm, thiab tuaj yeem tswj tau qee yam kev ruaj ntseg thiab kev lom zem ntawm cov pH sib txawv.
5. Thermochemical zog:
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 degree). Cov gases uas tsim thaum lub sij hawm thermal decomposition txheej txheem yog tsuas yog carbon dioxide, carbon monoxide, sulfuric acid gas, thiab lwm yam, yog li nws yog tsim nyog kom tsis txhob muaj kev kub ntxhov thaum lub sij hawm cia thiab siv.
Hauv kev xaus, Tirzepatide yog ib qho polypeptide molecule uas muaj ntau yam amino acid residues, thiab nws cov khoom siv tshuaj raug cuam tshuam los ntawm ntau yam. Tirzepatide muaj qee yam solubility, buffering peev xwm thiab acid-puag zog, thiab muaj peev xwm tswj tau lub sij hawm ntev stability nyob rau hauv tsim nyog tej yam kev mob. Txawm li cas los xij, nyob rau hauv cov xwm txheej xws li tus nqi pH tsawg dhau lossis siab dhau, qis dhau lossis kub dhau, thiab lub teeb, Tirzepatide tau yooj yim degraded thiab inactivated. Yog li ntawd, nws yog ib qho tsim nyog yuav tsum tau xyuam xim rau qhov cuam tshuam ntawm cov yam ntxwv no thaum khaws cia thiab siv Tirzepatide, thiab siv cov kev tiv thaiv tsim nyog los xyuas kom meej tias nws ua tau zoo thiab kev nyab xeeb.