Azure B: Ukungcola, I-Metabolite Nocwaningo

I-Azure B iyinhlanganisela ehlukile ngokwamakhemikhali ehlobene ne-methylene blue. Ingaba isivele ikhona kusampula ye-methylene blue, ivele ngemva kokuba umzimba usucubungule i-methylene blue, noma ihlinzekwe ngokwehlukana ukuze isetshenziswe ocwaningweni. Lezi zincazelo ziyahambisana. Zichaza ukuthi le nhlanganisela yavela kuphi nokuthi umcwaningi ubuza muphi umbuzo ngayo.

Lo mehluko ubalulekile lapho kuhunyushwa isitifiketi sokuhlaziywa noma isimangalo ngomsebenzi webhayoloji. Ukubiza i-Azure B nge-metabolite esebenzayo akusho ukuthi inani layo elingalawulwa ebhodleleni liyafiseleka. Ukuyibiza ngokungcola akusho ukuthi ayisebenzi ngokwebhayoloji.

Yini eshintshayo ngokwamakhemikhali?

I-Azure B iwumkhiqizo we-methylene blue osuswe iqembu elilodwa le-methyl. Iqembu elilodwa le-methyl elixhunywe ku-nitrogen lithathelwe indawo yi-hydrogen. Lobu budlelwano buchazwe engxoxweni yamakhemikhali naku-Figure 1 kokubuyekezwa kuka-Schirmer nozakwabo. Lolu ushintsho oluthile lwesakhiwo, hhayi nje elinye igama le-methylene blue.

Futhi kuhlukile ekuncishisweni kube yi-leucomethylene blue. Ukuncishiswa kushintsha isimo se-oxidation; i-N-demethylation ishintsha iqembu elixhunywe ku-nitrogen. Ukuphatha zombili lezi zinqubo njengokuthi nje “i-methylene blue iba engenambala” kususa umehluko odingekayo ukuze kuhunyushwe i-metabolism. Amagama amadayi ahlobene nawo adinga ukunakwa okufanayo: i-Azure A, i-Azure B, ne-methylene blue akuwona amagama angasetshenziswa ngokushintshana. Umhlahlandlela wezinhlanganisela ezinamagama afanayo uchaza le nkinga yokuqanjwa kwamagama.

Imvelaphi emithathu, imibuzo emithathu ehlukene

Umdwebo osesithombeni sekhava uhlukanisa izindlela ezintathu zophenyo. Ithebula lenza kucace ukuthi yiluphi ubufakazi obudingekayo endleleni ngayinye.

Lapho i-Azure B ingena khona ophenyweni	Umbuzo obuzwayo	Ubufakazi obuwusizo	Isiphetho esingalandeli
Isivele ikhona ezintweni zokuqala	Iqukethe ini le batch?	Indlela ehlukanisa futhi ilinganise i-Azure B, exhunyaniswe ne-lot kanye ne-specification esebenzayo	Inani elitholakele lithuthukisa imiphumela yomkhiqizo
Itholwa ngemva kokuchayeka ku-methylene blue	Yiziphi izinhlanganisela ezikhona ngokuhamba kwesikhathi?	Ukuhlaziywa kwezinto zokuqala kanye nezilinganiso zebhayoloji eziqhubekayo neziqinisekisiwe	Yonke i-molecule etholakele kumelwe ukuthi yakheke emzimbeni
Yengezwa njengenhlanganisela echazwe ngokwehlukana	Yenzani i-Azure B ngokwayo?	Ubunikazi, ubumsulwa, i-concentration, i-comparator, kanye ne-endpoint yocwaningo echazwe ngokucacile	Umphumela ofanayo uyenzeka ezigulini ezithatha i-methylene blue

Umugqa ophakathi uvame ukunganakwa. Uma i-Azure B yayikhona ezintweni ezinikeziwe, isampula yebhayoloji ethathwe kamuva ingaqukatha kokubili inhlanganisela eyayivele ikhona kanye nesanda kwakheka. Ukuqinisekisa ukuthi yakheka kudinga okungaphezu kokuthola i-peak ngemva kokuchayeka.

I-Azure B ezintweni zokuqala

Ucwaningo lomlando lokuhlaziya lunikeza isibonelo esiwusizo sokuthi kungani ingxenye encane ingaba nomthelela. Abacwaningi ababehlola i-iodination ye-methylene blue nge-HPLC–mass spectrometry bathola ukuthi umkhiqizo oyinhloko one-iodine wavela ekungcoleni kwe-Azure B. Amasampula e-methylene blue ayehlolwa ayequkethe cishe 7–8% Azure B. Ngaphansi kwezimo zabo zokusabela, i-Azure B ene-iodine yayiningi kakhulu kune-methylene blue ene-iodine.

Lawo maphesenti achaza izinto ezasetshenziswa kulolo cwaningo lwango-1999. Awamisi umkhawulo owamukelekayo wokungcola wanamuhla noma ukwakheka komkhiqizo womunye umhlinzeki. Ucwaningo lubonisa iphuzu elincane: inhlanganisela eningi kakhulu ekuqaleni akudingeki ukuthi ikhiqize umkhiqizo wokusabela omningi kakhulu.

Esitifiketini sokuhlaziywa, hlola umphumela othile we-Azure B, amayunithi, indlela yokuhlola, umkhawulo wokubika, kanye nobunikazi be-lot. I-total dye assay kanye nokuhlolwa kwe-related substances kuphendula imibuzo ehlukene. Umphumela othi “awutholakalanga” nawo udinga umkhawulo wokuthola noma wokubika ukuze uhunyushwe. Bheka indlela yokufunda isitifiketi sokuhlaziywa kwe-methylene blue ukuze uthole ukuhlolwa okubanzi kwedokhumenti.

I-Azure B eyakheka ngesikhathi se-metabolism

Ukutadisha i-metabolism kudinga ukuhlukaniswa ngokuhlaziya kwe-parent compound kanye ne-metabolite yayo. UKim nozakwabo basungula indlela ye-LC–MS/MS yokulinganisa zombili izinhlanganisela ku-plasma yamagundane. I-calibration range yabo yayingu-1–1,000 ng/mL ngenhlanganisela ngayinye, nomkhawulo ophansi wokulinganisa ongu-1 ng/mL kusetshenziswa 20 microlitres ye-plasma yamagundane. Babe sebeyisebenzisa le ndlela ocwaningweni lwe-pharmacokinetics lwamagundane.

Le mininingwane ibalulekile ngoba indlela yokulinganisa inomongo wesampula ethile. Ukuqinisekiswa ku-plasma yamagundane akusho ngokuzenzakalelayo ukunemba ku-plasma yomuntu, emchameni, noma kusixazululo sokuthengisa esigxilile. Le ndlela ibonisa ukuthi abacwaningi bangazilandela kanjani izinhlanganisela ezimbili ngokwehlukana; ayinikezi umthetho wokunikeza abantu.

Idatha yokuchayeka kubantu nayo ibonisa ukuthi kungani iphesenti elilodwa elingaguquki le-metabolite linganele. Isigaba se-clinical pharmacology selebula yomjovo we-methylene blue sibika ukwanda kokuchayeka ku-Azure B kubahlanganyeli abanokukhubazeka kokusebenza kwezinso. Ukuchayeka ngokuhamba kwesikhathi kubonisa ukwakheka, ukusatshalaliswa, kanye nokususwa, hhayi ukwakheka kwalokho okusebhodleleni kuphela. Indlela yokunikezwa, isikhathi sokuthatha isampula, kanye nendawo yomzimba elinganiswayo konke kubalulekile lapho kuqhathaniswa le miphumela.

Okuboniswa ukuhlolwa okuqondile

I-Azure B inomsebenzi we-pharmacology ongalinganiswa. Umbuzo owusizo kakhulu ukuthi isebenza kuphi, ngaphansi kwaziphi izimo.

Ocwaningweni lwango-2012 olusebenzisa i-recombinant human monoamine oxidase, i-Azure B yavimbela i-MAO-A nge-IC50 engu-11 nM, uma kuqhathaniswa no-70 nM we-methylene blue ngaphansi kwezimo ezifanayo. I-IC50 yi-concentration ehambisana nokuvinjelwa okungu-50% kulokho kuhlolwa. I-Azure B yaphinde yavimbela i-MAO-B, kodwa nge-concentration ephakeme kakhulu: 968 nM. Abacwaningi babika ukuvinjelwa okuguqukayo.

Le miphumela isekela ukuthi kungenzeka kube khona umthelela emisebenzini nasekusebenzisaneni kwebhayoloji, kuhlanganise nemibuzo mayelana ne-methylene blue nama-neurotransmitters. Ayisho ukuthi i-Azure B ingcono izikhathi eziyisithupha njengomuthi. Leso silinganiso sichaza ukuvinjelwa kwe-enzyme eyodwa ekuqhathanisweni okukodwa kokuhlolwa.

Ukuhlolwa okuhlukile kwe-human cholinesterase kubonisa umkhawulo walokho kuhlanganisa. Amanani e-IC50 e-Azure B ayengu-0.486 micromolar we-acetylcholinesterase kanye no-1.99 micromolar we-butyrylcholinesterase. I-methylene blue yayinamandla kakhulu kulezo zivivinyo ezifanayo, ngamanani angu-0.214 no-0.389 micromolar ngokulandelana. Ukuhleleka kwaguquka kuye ngethagethi yebhayoloji. Awukho kule miphumela owodwa, ngokuzimela, oqinisekisa ukuthuthukiswa kokucabanga noma ukwelashwa kwesifo i-Alzheimer's disease.

Kusukela emsebenzini wokuhlola kuye ekuhunyushweni komtholampilo

Ukuhlolwa ezilwaneni eziphelele kwengeza elinye izinga lobufakazi ngaphandle kokuxazulula ukusebenza komtholampilo. Ocwaningweni lwamagundane luka-Delport nozakwabo, i-Azure B yehlisa ukunganyakazi ekuhlolweni kwe-acute forced-swim, kanti i-methylene blue ne-imipramine kwakuphakathi kwama-comparator. Iphepha likubiza lokhu ngomphumela ofana nowe-antidepressant. Kwakuwucwaningo lokuziphatha kumagundane, hhayi ukuhlolwa okulinganisa ukululama ekucindezelekeni ezigulini.

Kunoma yisiphi isimangalo nge-Azure B, qala ngokukhomba izinto ezahlolwa. Bese ukhomba i-organism noma i-assay, ukuchayeka, kanye ne-endpoint. Ekugcineni, hlola ukuthi isiphetho sikhuluma ngokwakheka, i-metabolism, umsebenzi we-enzyme, ukuziphatha kwezilwane, noma inzuzo yomtholampilo. Lezi zinyathelo zivimbela ukuthi umphumela wekhwalithi ye-batch udidaniswe nesimangalo sokwelashwa, noma ukuthi indlela yokusebenza yocwaningo ididaniswe nomphumela osuqinisekisiwe ezigulini.