TR201809589A2 - DYEING METHOD PROVIDING EFFECT OF FABRIC - Google Patents
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Abstract
Dokunmuş veya örülmüş kumaşa efekt vermeyi sağlamak üzere bir boyama yöntemi ile ilgilidir. Yenilik olarak; bahsedilen kumaşın eni ve/veya boyu doğrultusunda burularak halat haline getirilmesi, bahsedilen halat haline getirilen kumaşın emdirme yöntemine göre boyanması işlem adımını içermektedir. Şekil 1It relates to a dyeing method to provide effect to the woven or knitted fabric. As innovation; and stitching said fabric in the width and / or length direction; Figure 1
Description
TARIFNAME PROTRANSDUZIN B, BIR GEN TRANSFERI GÜÇLENDIRICISI Açiklama Bu basvuru bir peptide, bu peptidi içeren bir ilaca, gen terapisinde kullaniin için bu peptide, genetik olarak tasarlanmis bir viral olusumla bir hücrenin enfeksiyonunu arttirmak için bir usule ve bu peptidin bir hücrenin bir virüsle enfeksiyonunun arttirilmasinda kullanimina iliskindir. DESCRIPTION PROTRANSDUZIN B, A GENE TRANSFER AMPLIFIER Explanation This application refers to a peptide, a drug containing this peptide, for use in gene therapy. peptide to increase infection of a cell by a genetically engineered viral and a method for increasing the infection of a cell with a virus of this peptide. related to its use.
Uygulanan usullerdeki muazzam ilerleme nedeniyle son yillarda genetik mühendisliginin önemi artti, çünkü sadece protein/peptit etken maddelerinin üretiminin degil, ayni zamanda hücrelerin bir laboratuvar araci olarak stabil genlerle transfeksiyonunun ve sonuçta gen kusurlari için bir çare olarak genlerin hücrelere dâhil edilmesinin çok sayida hastaligin tedavisi için çok yararli olacagi öngörülebilmektedir. Due to the tremendous progress in the methods applied, genetic engineering has become more popular in recent years. increased in importance, because not only the production of protein/peptide active substances, but also transfection of cells with stable genes as a laboratory tool and ultimately gene The incorporation of genes into cells as a remedy for It can be predicted that it will be very useful for the treatment.
Ilk beseri genlerin klonlanmasindan ve rekombinant üretimden bu yana, gen transferi usulleri artan veriinlilikle sürekli ilerledigi için spesifik hücre fonksiyonlarini degistirmek için genetik malzemenin dâhil edilmesi temel ve uygulamali biyolojik-tibbi arastirmanin vazgeçilmez bir araci haline geldi. Gen dâhil etmenin çok sayida usulü optimizasyona yol açti. Baslangiçta çok yavas olmakla birlikte, uzun yillar boyunca uygun deneyler toplandi. islevinin açiklanmasindan bile önce, F. Griffith 1920ilerin sonlarina dogru deneylerde apatoj enik Pneumococcus suslarini patoj enlere transforme etmeyi basardi. Bu transformasyon sansli bir durumdan kaynaklanmisti, çünkü pnömokoklar nadir bir DNA alimi kabiliyetine sahipti. DNA”nin spesifik bir sekilde prokaryotlar içine dâhil edilmesi, digerlerinin yani sira J. Lederberg, M. Delbrück ve S. Luria tarafindan fajlar vasitasiyla saglandi, buna transdüksiyon adi verildi. Hücre kültürünün saglanmasi, ökaryotik hücrelerin in vitro kosullar altinda kültürlenmesiyle birlikte, transfeksiyon için bir dizi fiziksel ve kimyasal usul gelistirildi. Daha sik kullanilan, ama daha pahali ekipman gerektiren fiziksel usuller arasinda, l980”lerde ve bugün hâlâ alisilageldik olan kalsiyum fosfat çökeltme usulü veya katyonik lipidlere ve katyonik polimerlere dayanan, 1990,larin baslarinda yaygin olan usuller gibi daha basit uygulanabilen kimyasal usullerle rekabet eden elektroporasyon ve mikroenj eksiyon yer alir. Ancak bu usullerin kullanimi her zaman hücrelere veya DNA°ya dayaniyordu. Ayrica hücrelere dâhil edilen DNA genellikle ekstrakromozomaldi (geçici transfeksiyon) ve dolayisiyla yavru hücrelere aktarilmiyordu. Ancak, fajlarin (örnegin lainbda faj) DNAllarini konakçi genomuna entegre ettigi biliniyordu (profaj, lizoj enik enfeksiyon yolu). Buradan, “Retrovirüslerin gen vektörleri olarak saptanmasi”na (Doehmer ve digerleri ve Tabin ve digerleri) sadece bir adim vardi (1981/1982). Virüsler türe spesifik ve organa/dokuya spesifiktir, bu nedenle bütün virüsler bütün (ökaryotik) hücreleri enfekte etmez. Viral zarftaki degisiklikler (glikoproteinlerin degisimi, psödotip virüsler) ve çogunlukla katyonik olan peptitlerin ilave edilmesinin transdüksiyon etkinligini arttirdi gi varsayilmaktadir. Since the cloning of the first human genes and recombinant production, gene transfer has been modifying specific cell functions as their methods are constantly advancing with increasing efficiency inclusion of genetic material for basic and applied biological-medical research It has become an indispensable tool. Multiple methods of gene inclusion lead to optimization. opened. Very slowly at first, but over many years suitable experiments were collected. your function Even before its disclosure, F. Griffith was apathogenic in experiments in the late 1920s. He succeeded in transforming Pneumococcus strains into pathogens. This transformation is lucky was caused by a condition, because pneumococci have a rare ability to take up DNA. he had. The specific incorporation of DNA into prokaryotes provided via phages by J. Lederberg, M. Delbrück, and S. Luria, respectively. called transduction. Providing cell culture, eukaryotic cells in vitro conditions, a number of physical and chemical processes are required for transfection. method was developed. Physical methods that are used more often but require more expensive equipment the calcium phosphate precipitation method, which was customary in the 1980s and still today, or widespread in the early 1990s, based on cationic lipids and cationic polymers. electroporation, which competes with simpler applicable chemical methods such as microinjection is included. However, the use of these methods does not always involve cells or DNA. was based. In addition, the DNA incorporated into cells was often extrachromosomal (temporary transfection) and therefore not transferred to daughter cells. However, phages (for example lainbda phage) were known to integrate their DNA into the host genome (prophage, lysogenic route of infection). From here to “Detection of retroviruses as gene vectors” (Doehmer and others and Tabin et al) there was only one step (1981/1982). Viruses are species specific and is organ/tissue specific, so all viruses infect all (eukaryotic) cells it doesn't. Changes in the viral envelope (change of glycoproteins, pseudotype viruses) and The addition of mostly cationic peptides increased the transduction efficiency. is assumed.
Virüs parçaciklari aliminin ilk güçlendiricileri HIV çalismasinda dikkat çekti. Bir spesifik hücre testi vasitasiyla in vitro enfeksiyon analizleri sirasinda, HIV füzyonunun kan filtrat peptitleriyle inhibisyonu gözlemlendi (Münch ve digerleri, VIRIP). The first enhancers of virus particle uptake were noted in the HIV study. a specific During in vitro infection assays by cell test, the blood filtrate of HIV fusion peptides were observed (Münch et al., VIRIP).
Proteinlerin, sasirtici bir sekilde dogal olarak olusan bu fragmanlarinin insan sperminde, amiloid fibriller olarak karakterize edilen güçlendiriciler olarak lifli yapilar - “Virüs Enfeksiyonunun Sperm türevli Güçlendiricisi” (SEVI) - olusturdugu gözlemlendi. Bu nanofibriller virüslerin hedef hücrelerine kenetlenmesini destekleyerek, viral enfeksiyon hizini onun birkaç kati arttirir. Surprisingly, these naturally occurring fragments of proteins in human sperm fibrous structures as reinforcers characterized as amyloid fibrils - “Virus Sperm-derived Enhancer (SEVI) - was observed. This nanofibrils support the docking of viruses to their target cells, thereby reducing viral infection. increases its speed several times.
Bu, temel arastirma ve gelecekteki muhtemel terapötik uygulamalar için retroviral gen transferini gelistirmek için kullanildi. Böylece, gen terapisi için kullanilan lentiviral ve gama-retroviral vektörlerin beseri T hücreleri, servikal karsinom hücreleri, lösemi hücreleri, hematopoietik kök hücreler ve embriyonik kök hücreler gibi farkli hücre tipleri için SEVI proteinin varliginda çok daha yüksek gen transferi hizi gösterdigi ortaya SEVl ve seminogelin gibi baska güçlendiricilerin gelistirilmesine yönelik çalismalar, viral zarf proteinlerinden peptitlerin de transfeksiyon güçlendiricileri olarak uygun olabilecegi varsayiinina yol açti ve bu varsayim sasirtici bir sekilde beklenmedik ölçüde büyük bir basari kazandi (Maral Yolamanova, Nature Nanotechnology, Cilt 8, No. 2, sayfa 130-136). This is a retroviral gene for basic research and possible future therapeutic applications. used to improve the transfer. Thus, lentiviral and human T cells, cervical carcinoma cells, leukemia of gamma-retroviral vectors different cell types such as hematopoietic stem cells and embryonic stem cells It was revealed that the SEVI protein showed a much higher rate of gene transfer in the presence of Studies to develop other enhancers, such as SEV1 and seminogelin, Peptides from envelope proteins may also be suitable as transfection enhancers. led to the assumption that, surprisingly, this assumption had an unexpectedly large was successful (Maral Yolamanova, Nature Nanotechnology, Vol. 8, No. 2, pp. 130-136).
Böylece, örnegin farkli konsantrasyonlarda (1-100 ug/m1)pr0transduzin A (es anlamlisi EF-C) ile önceden inkübe edilen HIVllerin raportör hücrelerle onun birkaç kati artan bir enfeksiyon gösterdigi ortaya konulabildi. Etki mekanizmasi olarak, EF-C”nin virüsleri baglayabilen ve konsantre edebilen ve dolayisiyla virüslerin hücre içine girisini arttirabilen fibriler yapilar olusturdugu varsayildi. Viral parçaciklarla enfeksiyonun yani sira, EF-C gen terapisinde uygulanan çok çesitli beseri hücre tiplerinde (T hücreleri, gliyal hücreler, fibroblastlar, hematopoietik kök hücreler) yüksek verimlilikle lentiviral ve retroviral parçaciklarin transdüksiyonunu arttirmaktadir (Jan Münch ve digerleri, Nature iliskindir. Thus, for example, different concentrations (1-100 µg/m1) of pr0transduzin A (synonymous) HIVls pre-incubated with EF-C) showed a several-fold increased association with reporter cells. infection could be demonstrated. As a mechanism of action, viruses of EF-C able to bind and concentrate and therefore increase the entry of viruses into the cell. fibrillar structures were assumed. Besides infection with viral particles, EF-C in a wide variety of human cell types (T cells, glial cells, fibroblasts, hematopoietic stem cells) lentiviral and retroviral with high efficiency increases the transduction of particles (Jan Münch et al., Nature is related.
Yukarida açiklandigi gibi gen teknolojisinin artan Önemi nedeniyle, gen transferinin daha etkili güçlendiricileri istenmektedir. Bulusun amaci gelistirilmis bir gen transferi güçlendiricisi sunmaktir. As explained above, due to the increasing importance of gene technology, gene transfer has become more effective enhancers are required. The aim of the invention is an improved gene transfer to offer a booster.
Sasirtici bir sekilde asagidaki diziye sahip bir peptidin bulusun amacina ulastigi tespit edilmistir: X-Glu-Cys-Lys-lle-Lys-Gln-Ile-lle-Asn-Met-Trp-Gln (DIZI ID NO: 1), burada X-Glu grubu amino asit piroglutamik asidi temsil eder Sasirtici bir sekilde sulu çözeltide protransduzinin stabilitesindeki büyük bir artisa yol açanin, in vitro (hücresel etkiler, özellikle enzimlerin varligi olmadan) piroglutamik asitle N-terminal ucun modifikasyonu oldugu tespit edildi. Bu, Sekil 1”de gösterilen sonuçlardan açikça görülmektedir. depolama üzerine protransduzin A için sonuçlar, ayni kosullar altinda protransduzin B için sonuçlarla karsilastirilmaktadir. Protransduzin A”nin 13 gün 4°C3de depolama sonucunda yarisina kadar bozundugu açiktir, protransduzin B ise ayni depolama kosullari altinda hemen hemen hiç bozunmadi (piklerin yüksekligi numunede bulunan bilesenlerin konsantrasyonuna karsilik gelmektedir). digerleri amiloid peptitlere, özellikle piroglutamatla modifiye edilmis amiloid polipeptitlere iliskin son durumu bildirmektedirler. Bu amiloid polipeptitler çok sayida ainino aside sahiptir ve temel olarak, bulusa uygun olanlarin da dâhil oldugu kisa zincirlerle karsilastirilabilir degildir. Bu arada, bu kisa inceleme, protransduzinin stabilitesinin bulusa göre gelistirildigi kosullarla, yani in vitro kosullarla hiçbir sekilde karsilastirilabilir olmayan bir sekilde, enzimlerin varliginda in Vivo kosullar altinda gerçeklesen hücresel olaylara iliskindir, Bulusa uygun peptit bir ilaç olarak da kullanilabilir. Surprisingly, it was found that a peptide with the following sequence fulfilled the purpose of the invention. has been: X-Glu-Cys-Lys-lle-Lys-Gln-Ile-lle-Asn-Met-Trp-Gln (SEQ ID NO: 1), where X-Glu group represents amino acid pyroglutamic acid Surprisingly, it resulted in a large increase in the stability of protransdusine in aqueous solution. In vitro (without cellular effects, especially the presence of enzymes) of azacine with pyroglutamic acid It was determined that there was a modification of the N-terminal end. This is from the results shown in Figure 1 is clearly visible. The results for protransdusin A upon storage, under the same conditions, for protransdusin B. are compared with the results. After 13 days of storage of Protransduzin A at 4°C It is clear that it is degraded by half, while protransdusin B is stored under the same storage conditions. almost never decayed (the height of the peaks corresponds to that of the components present in the sample). corresponding to its concentration). others to amyloid peptides, particularly pyroglutamate-modified amyloid They report the latest situation regarding polypeptides. These amyloid polypeptides are numerous. It has ainino acid and is basically short-lived, including those according to the invention. not comparable to chains. Incidentally, this brief review under the conditions under which its stability was developed according to the invention, i.e. in vitro. not comparable under in vivo conditions in the presence of enzymes. relates to cellular events that take place, The peptide according to the invention can also be used as a medicine.
Bulus ayrica bulusa uygun peptidin, gen terapisiyle tedavi edilebilen hastaliklari tedavi etmek için gen terapisinde kullanimina iliskindir. The invention also provides that the peptide according to the invention can treat diseases treatable by gene therapy. for its use in gene therapy.
Bulus ayrica bir hücrenin bir virüs tarafindan enfeksiyonunu arttirmak için, asagidaki asamalari içeren bir usule iliskindir: - bir organik çözücü içinde erimis olarak istem lie uygun peptidi temin etmek; - peptidin çözünmez yigisimlarini olusturmak için peptidi sulu bir çözeltiye ilave etmek; - yukaridaki son asamada elde edilen çözeltiyi karistirmak; ve - hücreleri kültürleinek. The invention also uses the following methods to increase the infection of a cell by a virus. It relates to a procedure that includes steps: - providing the peptide according to claim dissolved in an organic solvent; - adding the peptide to an aqueous solution to form insoluble aggregates of the peptide; - mixing the solution obtained in the last step above; and - culture the cells.
Bu bulus ayrica bulusa uygun peptidin bir hücrenin bir virüsle enfeksiyonunu arttirmak için kullanimina iliskindir. Son olarak, bulusa uygun peptidi içeren bir kit de sunulmaktadir. The invention also provides that the peptide according to the invention can be used to increase infection of a cell with a virus. for its use. Finally, a kit containing the peptide of the invention is also available. is offered.
Bulusa uygun peptit (protransduzin B) Merrifieldie uygun usulle Fmoc korumali amino asitlerle hazirlanabilir. Peptide of the invention (protransduzin B) Fmoc protected amino by the method according to Merrifieldie can be prepared with acids.
Bu usul Fmoc korumali türevlerle, yani (9-Ilorenilmetoksikarbonil) korumali amino asitlerle, Merrifield prensibine göre asamali bir kati faz sentezinde, özellikle sentezleyici ABI-433 üzerindeki bir kati destek olarak önceden Fmoc-L-glutamin (0.59 mmol/g, 100- 200 elek numarasi) yüklenmis bir Wang reçinesi üzerinde isler. This method is used with Fmoc protected derivatives, i.e. (9-Ilorenylmethoxycarbonyl) protected amino acids, in a stepwise solid phase synthesis according to the Merrifield principle, especially synthesizing As a solid supplement on ABI-433, previously Fmoc-L-glutamine (0.59 mmol/g, 100- 200 mesh) on a loaded Wang resin.
Tipik olarak on kat molar fazlalikta kullanilan Fmoc-L-amino asitlerin aktivasyonu oda sicakliginda N-metil-Z-pirolidinon (NMP) içinde ve 2 M diizopropiletilamin (DIEA) ilaveleriyle [(2-( 1H-benzotriazol-1-il)-l,1,3,3- tetrametiluronyuin heksaflorofosfat] (HBTU, 100 mmol/l) ile yapilir. Activation of Fmoc-L-amino acids, typically used in a ten-fold molar excess in N-methyl-Z-pyrrolidinone (NMP) at temperature and [(2-( 1H-benzotriazol-1-yl)-1,1,3,3-) with additions of 2 M diisopropylethylamine (DIEA) tetramethyluroniin hexafluorophosphate] (HBTU, 100 mmol/l).
Münferit açilasyon reaksiyonlari 45 dakika sürer ve %20 piperidinle Fmoc korumasinin kaldirilmasi 15 dakika sürer. Individual acylation reactions take 45 minutes and Fmoc protection is reduced with 20% piperidine. It takes 15 minutes to uninstall.
Sentez için asagidaki amino asit türevleri ve iliskili ortogonal asitle yarilabilir yan zincir koruyucu gruplari kullanilir: Fmoc-L-Asn(Trt), Fmoc-L-Cys(Trt), L-pGlu, Fmoc-L-Gln(Trt), Fmoc-L-Ile, Fmoc-L- Lys(Boc), Fmoc-L-Met ve Fmoc-L-Trp(Boc). For synthesis, the following amino acid derivatives and the associated orthogonal acid cleavable side chain protecting groups are used: Fmoc-L-Asn(Trt), Fmoc-L-Cys(Trt), L-pGlu, Fmoc-L-Gln(Trt), Fmoc-L-Ile, Fmoc-L- Lys(Boc), Fmoc-L-Met and Fmoc-L-Trp(Boc).
Reçine desteginin %94 trifloroasetik asit (TF A), %3 etanditiol (EDT) ve %3 minerali giderilmis suyla peptidil reçineden yarilmasindan sonra, islenmemis peptit soguk ter-bütil metil eter içinde çökeltilir, islenmemis peptit santrifujden geçirilerek bir pelet elde edilir ve üst faz atilir. 94% trifluoroacetic acid (TFA), 3% ethanedithiol (EDT) and 3% mineral of the resin backing After cleavage from the peptidyl resin with decontaminated water, the uncured peptide is cold tert-butyl precipitated in methyl ether, centrifuged the crude peptide to obtain a pellet, and upper phase is discarded.
Islenmemis peptidin sonraki kromatografik saflastirilmasi gradyan elüsyonuyla preparatif bir sekilde yapilir. Subsequent chromatographic purification of the crude peptide is preparative by gradient elution. done in one go.
EP 2 452 947 A1”e uygun protransduzin A ile protransduzin B arasindaki fark, sentetik L- piroglutamik asidin (pGlu) protransduzin Blde sentetik L- glutamin (Gln) yerine N-terminal olarak sokulmasinda yatar. Orijinal glutamin bir laktam olusturmak için halka kapanisiyla modifiye edilir. According to EP 2 452 947 A1 the difference between protransdusin A and protransdusin B is that synthetic L- N-terminal of pyroglutamic acid (pGlu) instead of synthetic L-glutamine (Gln) in protransdusin Bl lies in its insertion. The original glutamine is combined with a ring closure to form a lactam. is modified.
Saflastirma: Preparatif ayirma: Saflastirma Gilson company,den bir HPLC üzerinde uygulanir. UV/VlS detektörü Kronwald companyidendir ve ayrilina 230 nm dalga uzunlugunda saptanir. Akis hizi 40 ml/dakikadir. Purification: Preparative separation: Purification is carried out on an HPLC from Gilson company. UV/VLS The detector is from the Kronwald company and is detected at a wavelength of 230 nm apart. Flow rate is 40 ml/minute.
Kolon bir Waters Prep-Pak C18 kartustur (47 x 300 mm). The column is a Waters Prep-Pak C18 cartridge (47 x 300 mm).
Elüsyon maddesi A: Minerali giderilmis su içinde %01 TFA; elüsyon maddesi B: %80 asetonitril ve %20 minerali giderilmis su içinde %0.] TFA. Eluent A: 01% TFA in demineralised water; eluent B: 80% 0.%] TFA in acetonitrile and 20% demineralised water.
Protransduzin B için gradyan 40 dakikada %3 5-%55 elüsyon maddesi B, yani dakikada Protransduzin B %40 elüsyon maddesi B”de elute olur ve 0.5 ila 1 dakikalik birkaç fraksiyon halinde toplanir. Analitik olarak temiz fraksiyonlar havuzlanir ve liyofilize edilir. For protransduzin B, the gradient is 3% to 55% eluent B over 40 min, i.e. min. Protransduzin B elutes in 40% eluent B and takes several minutes of 0.5 to 1 min. collected in fractions. The analytically clean fractions are pooled and lyophilized.
Uygulama için islem: Liyofilizasyon: Dondurarak kurutma için, teknik verileri asagidaki gibi belirlenen, Christ company,nin kondansatör performansi maksimum 45 kg/24 saat; vakum pompasinin nihai kismi basinci, gaz dengeleyici kullanilarak/kullanilmadan 1 x 10-3/10'4 mbar; dondurarak kurutma verileri (birim gaz dengeleyiciyle manuel olarak çalistirilir): nihai kismi basinç 1 x 10*2 mbar; buz kondansatör sicakligi -50°C; raf sicakligi +15°C; raf isitmasinin çalisma noktasi 0.5 mbar; dondurarak kurutma süresi 3 güne kadar. Procedure for the application: Lyophilization: For freeze drying, the technical data of the Christ company, whose technical data are determined as follows, capacitor performance maximum 45 kg/24 hours; final pressure of the vacuum pump, 1 x 10-3/10'4 mbar with/without gas stabilizer; freeze drying data (unit operated manually with gas stabilizer): final partial pressure 1 x 10*2 mbar; ice condenser temperature -50°C; shelf temperature +15°C; shelf heating operating point 0.5 mbar; freeze drying time up to 3 days.
Hücrelerin protransduzin B ile transdüksiyonu 0.5 mg protransduzin B,yi 50 ul DMSO içinde eritin. Daha sonra çözeltiye 450 ul PBS ilave edin, 3 dakika içinde Iibriller olusur. Bu stok çözeltisini (l mg/ml) vektörlere ilave ederek 25 ug/ml protransduzin B konsantrasyonu elde edin. Çözeltiyi 1 dakika vorteksledikten sonra 5 dakika 5000 g°yle santrifüjden geçirin. Üst fazi atin ve peleti biraz PBS içinde süspansiyon haline getirin ve hücrelere ilave ediln. Hücreler 2 gün bir inkübatörde inkübe edilir. Transduction of cells with protransdusin B Dissolve 0.5 mg of protransduzin B in 50 µl of DMSO. Then add 450 µl of PBS to the solution. add, within 3 minutes ibrils will form. Add this stock solution (1 mg/ml) to the vectors. achieve a concentration of 25 µg/ml protransdusin B. 1 minute of solution After vortexing, centrifuge at 5000 g for 5 minutes. Discard the top phase and a little bit of the pellet Suspend in PBS and add to cells. Cells every 2 days incubated in the incubator.
Transdüksiyon hizi protransduzin B ile önemli ölçüde artar. Hücrelerin %96`ya kadari protransduzin B ile transdüksiyondan geçirilebilir. The rate of transduction is significantly increased by protransdusin B. Up to 96% of cells It can be transduced with protransduzin B.
Dosya No. :EP-17983 Resim Adedi: 1 EP 2 992 004 B1 Sayfa No : 1 -20°C PTD-A h PTD-B -20°C -20°C t=13 t=13 eîririiwk k( Md ::lafin ;im [Ml'MÜQQMQgßtmmm Hanim? i ”il”Hl › ji'Q'Qiiirigq Testten önce katalizörler, ZSOOC'ta azot içinde kurutmayi, daha sonra ayni sicaklikta 1 saat boyunca hidrojen sülfür ile sülfidasyonu içeren bir birinci asamadan, ardindan 350°C`ta 1 saat boyunca H2/H2S ile indirgeme / sülfidasyon asamasindan olusan bir islem vasitasiyla yerinde aktive edilmistir. File number. :EP-17983 Number of Paintings: 1 EP 2 992 004 B1 Page No: 1 -20°C PTD-A h PTD-B -20°C -20°C t=13 t=13 eîririiwk k( Md ::lafin ;im [Ml'MÜQQMQgßtmmm Woman? i ”il”Hl › ji'Q'Qiiirigq Before testing, the catalysts were to dry in nitrogen at ZSOOC, then at the same temperature for 1 hour. a first step, which includes sulfidation with hydrogen sulfide, followed by 1 hour at 350°C. in situ through a process consisting of a reduction / sulfidation step with H2/H2S throughout is activated.
Sonuçlar, asagidaki Tablo 2'de verilmistir. The results are given in Table 2 below.
TABLO 2 : Katalitik testlerin sonuçlari Molar seçicilikler (%) Örnekler Katalizör COz/CH3 SH Orani CH3SH cos CO2 Tablo 2'de verilen sonuçlar, bulusa uygun katalizörlerin (Örnek 1 ve 2), bilinen teknigin destekleri (silis: örnek 3 veya alüinin: örnek 4 ve 5) üzerindeki katalizörlere kiyasla öneinli ölçüde daha düsük C02 (istenmeyen ürün) seçicilikleri sagladiklarini göstermektedir. TABLE 2 : Results of catalytic tests Molar selectivities (%) Examples Catalyst CO2/CH3 SH Ratio CH3SH cos CO2 The results given in Table 2 show that the catalysts according to the invention (Examples 1 and 2) are prior art. significant compared to catalysts on supports (silica: example 3 or alumina: examples 4 and 5) significantly lower C02 (undesirable product) selectivities.
Seçicilikler, karbon monoksidin izo-dönüsümü ile karsilastirilmistir; bu dönüsüm, katalizörün spesifik yüzey alani mz'si basina ifade edilmistir. Selectivities are compared with the isoconversion of carbon monoxide; this transformation is the catalyst expressed per mz of specific surface area.
Katalizör 1 ve 4 ile elde edilen sonuçlarin karsilastirilmasiyla, oran açisindan %30'1uk bir gelisme gözlemlenmistir; bu gelisme, destek olarak hidroksiapatitin seçilmesine baglidir. Comparing the results obtained with catalysts 1 and 4, a 30% reduction in ratio improvement has been observed; This improvement is due to the selection of hydroxyapatite as the support.
Bulusa uygun Örnek 2 ile Örnek 3 ve 5 karsilastirildiginda ayni gözlem yapilmistir. The same observation was made when comparing Example 2 according to the invention with Examples 3 and 5.
Böylece bir ikincil reaksiyon sonucu üretilen karbondioksite göre metil merkaptan için artan bir seçicilik tespit edilmistir. Thus, the increase for methyl mercaptan relative to the carbon dioxide produced as a result of a secondary reaction selectivity was detected.
Bu seçiciligin, bilinen teknikte açiklandigi gibi tellür oksit, nikel oksit veya demir oksit gibi bir promotörün yardiini olmaksizin elde edildigi dikkate alinmalidir. This selectivity may be due to such selectivity as tellurium oxide, nickel oxide or iron oxide as described in the prior art. It should be noted that it was achieved without the aid of a promoter.
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