Name: scaled-up preparation of an intermediate of upatinib, act051-3
Uploaded: 2023-04-07T14:00:00
Description: Watch this Scientific Journal Video about Scaled-Up Preparation of an Intermediate of Upatinib, ACT051-3 at JoVE.com

There are no acknowledgments to mention here.

1. Pilot scale synthesis of compounds ACT051-2 and ACT051-3
<ol>
	<li>Synthesis of 2-bromo-5-tosyl-5H-pyrrolo[2,3-b]pyrazine (ACT051-2)
	<ol>
		<li>In a round-bottom flask, dissolve 50.0 g of the compound 2-bromo-5H-pyrrolo[2,3-b]pyrazine (ACT051-1; 0.25 M) in 15 mL of N,N-dimethylformamide (DMF; 3 V).</li>
		<li>Add 65.3 g of diisopropylethylamine (DIPEA; 0.51 M) to the reaction solution under nitrogen protection (provide a gas and moisture barrier) and cool the temperature to 0-5 &#176;C through a co...

<ol>
	<li>Kerschbaumer, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Points+to+consider+for+the+treatment+of+immune-mediated+inflammatory+diseases+with+Janus+kinase+inhibitors:+a+systematic+literature+research.">Points to consider for the treatment of immune-mediated inflammatory diseases with Janus kinase inhibitors: a systematic literature research.</a> RMD Open. 6 (3), e001374 (2020).</li><li>Fragoulis, G. E., Brock, J., Basu, N., McInnes, I. 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G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=JAK+selectivity+and+the+implications+for+clinical+inhibition+of+pharmacodynamic+cytokine+signalling+by+filgotinib,+upadacitinib,+tofacitinib+and+baricitinib.">JAK selectivity and the implications for clinical inhibition of pharmacodynamic cytokine signalling by filgotinib, upadacitinib, tofacitinib and baricitinib.</a> Annals of the Rheumatic Diseases. 80 (7), 865-875 (2021).</li><li>Rozema, M. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+a+scalable+enantioselective+synthesis+of+JAK+inhibitor+upadacitinib.">Development of a scalable enantioselective synthesis of JAK inhibitor upadacitinib.</a> Organic Process Research &amp; Development. 26 (3), 949-962 (2022).</li><li>Wynn, J. P., Hanchar, R., Kleff, S., Senyk, D., Tiedje, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biobased+technology+commercialization:+the+importance+of+lab+to+pilot+scale-up.">Biobased technology commercialization: the importance of lab to pilot scale-up.</a> Metabolic Engineering for Bioprocess Commercialization. , 101-119 (2016).</li><li>Tang, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Influenza virus replication inhibitor and use thereof. , (2020).</li><li>Ren, Q., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inhibitors+of+influenza+virus+replication+and+uses+thereof.">Inhibitors of influenza virus replication and uses thereof.</a> Center for Biotechnology Information. , (2020).</li><li>Van Epps, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Design+and+synthesis+of+tricyclic+cores+for+kinase+inhibition.">Design and synthesis of tricyclic cores for kinase inhibition.</a> Bioorganic &amp; Medicinal Chemistry Letters. 23 (3), 693-698 (2013).</li><li>Paul, F., Patt, J., Hartwig, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Palladium-catalyzed+formation+of+carbon-nitrogen+bonds.+Reaction+intermediates+and+catalyst+improvements+in+the+hetero+cross-coupling+of+aryl+halides+and+tin+amides.">Palladium-catalyzed formation of carbon-nitrogen bonds. Reaction intermediates and catalyst improvements in the hetero cross-coupling of aryl halides and tin amides.</a> Journal of the American Chemical Society. 116 (13), 5969-5970 (1994).</li><li>Zhou, T., Ji, C. L., Hong, X., Szostak, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Palladium-catalyzed+decarbonylative+Suzuki-Miyaura+cross-coupling+of+amides+by+carbon-nitrogen+bond+activation.">Palladium-catalyzed decarbonylative Suzuki-Miyaura cross-coupling of amides by carbon-nitrogen bond activation.</a> Chemical Science. 10 (42), 9865-9871 (2019).</li><li>Sain, S., Jain, S., Srivastava, M., Vishwakarma, R., Dwivedi, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Application+of+palladium-catalyzed+cross-coupling+reactions+in+organic+synthesis.">Application of palladium-catalyzed cross-coupling reactions in organic synthesis.</a> Current Organic Synthesis. 16 (8), 1105-1142 (2019).</li><li>Takale, B. S., Kong, F. Y., Thakore, R. 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The reaction conditions of synthesis, including reaction temperature, time, the selection of reaction reagents, and the ratio of materials, affect the feasibility of the reaction, yields, purity, and production cost, especially for scale-up production.
In the laboratory synthesis of ACT051-2, TsCl in liquid form (dissolved in DMF; step 1.1.3) can be used to in the reaction; however, it is not suitable for pilot scale-up synthesis or industrial production, as using liquid TsCl for this reaction...

Upatinib has become a globally popular Janus kinase 1 (JAK1) inhibitor for treating immune disorders in recent years1,2. This drug has demonstrated significant therapeutic effects on psoriaticarthritis (PsA)3,4, rheumatoid arthritis (RA)5,6,7, and atopic dermatitis (AD)8,9. Additionally, due to its high selectivity10, Upatinib has a wide range of clinical applications. Tert-butyl (5-tosyl-5h-pyrolo [2,3-b] pyrazin-2-yl) carbamate (ACT051-3) is an important intermediate of Upatinib. Its main structural components are the pyrrole ring and the pyrazine ring, which can be used in the preparation of new nitrogen-containing tricyclic kinase inhibitors for the treatment of immune and tumor diseases11.
Pilot scale-up is a medium-sized scale-up (50x-100x) of the process route and conditions determined by the laboratory pilot study, followed by process testing, industrial investigation, and optimization to determine the best industrial production and operating conditions12.
At present, the lab synthesis routes for this intermediate compound (ACT051-3) have been reported, but they have only been performed on a small scale due to problems of low yield, complex reactions, and high equipment requirements, which still have a lot of room to be optimized11,13,14,15. However, no process route has been reported for pilot scale-up and industrial production of the intermediate compound ACT051-3 at present.
Therefore, in this study, we investigated the pilot scale-up and production route of the compound ACT051-3, with reference to the better reported laboratory synthetic routes. Compared with the original laboratory synthesis route, many appropriate adjustments and improvements were made to the reaction conditions, and other factors that may affect the reaction results were investigated. Finally, the most suitable process parameters for the optimal route were identified, and we obtained a process route that is simple to operate, low cost, and environmentally friendly, which is suitable for the pilot scale-up and production of ACT051-3.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>2-bromo-5H-pyrrolo[2,3-b]pyrazine</td>
			<td>Nanjing Cook Biotechnology Co., Ltd.</td>
			<td>19120110</td>
			<td></td>
		</tr>
		<tr>
			<td>1,4-dioxane</td>
			<td>Liaoning cook Biotechnology Co., Ltd</td>
			<td>General Reagent</td>
			<td></td>
		</tr>
		<tr>
			<td>1H NMR</td>
			<td></td>
			<td>Bruker AVIII 500</td>
			<td></td>
		</tr>
		<tr>
			<td>37% chloride acid molecular grade</td>
			<td>NEON</td>
			<td>02618 NEON</td>
			<td></td>
		</tr>
		<tr>
			<td>4-toluenesulfonyl chloride (TsCl)</td>
			<td>Nanjing Cook Biotechnology Co., Ltd.</td>
			<td>AR A2010137</td>
			<td></td>
		</tr>
		<tr>
			<td>Anti-Chicken IgY (H+L), highly cross-adsorbed, CF&#160;488A antibody produced in donkey</td>
			<td>Sigma-Aldrich</td>
			<td>SAB4600031</td>
			<td></td>
		</tr>
		<tr>
			<td>Anti-mouse IgG (H+L), F(ab&#8242;)2</td>
			<td>Sigma-Aldrich</td>
			<td>SAB4600388</td>
			<td></td>
		</tr>
		<tr>
			<td>BD FACSCanto II</td>
			<td>BD Biosciences</td>
			<td>BF-FACSC2</td>
			<td></td>
		</tr>
		<tr>
			<td>BD FACSDiva CS&#38;T research beads (CS&#38;T research beads)</td>
			<td>BD Biosciences</td>
			<td>655050</td>
			<td></td>
		</tr>
		<tr>
			<td>BD FACSDiva software 7.0</td>
			<td>BD Biosciences</td>
			<td>655677</td>
			<td></td>
		</tr>
		<tr>
			<td>Bovine serum albumin</td>
			<td>Sigma-Aldrich</td>
			<td>A4503</td>
			<td></td>
		</tr>
		<tr>
			<td>Centrifuge 5702 R</td>
			<td>Eppendorf</td>
			<td>Z606936</td>
			<td></td>
		</tr>
		<tr>
			<td>Circulating water vacuum pump</td>
			<td>Guangzhou Zhiyan Instrument Co., Ltd</td>
			<td>SHZ-D(<img alt="Equation 1" src="/files/ftp_upload/64514/64514eq01.jpg" style="margin: auto;" />)</td>
			<td></td>
		</tr>
		<tr>
			<td>CML latex, 4% w/v</td>
			<td>Invitrogen</td>
			<td>C37253</td>
			<td></td>
		</tr>
		<tr>
			<td>Diatomite</td>
			<td>Guangzhou Qishuo Chemical Co., Ltd.</td>
			<td>/</td>
			<td></td>
		</tr>
		<tr>
			<td>Double cone rotary vacuum dryer</td>
			<td>Jiangsu Yang-Yang Chemical Equipment Plant Inc</td>
			<td>SZE-500T</td>
			<td></td>
		</tr>
		<tr>
			<td>enamel kettle</td>
			<td>Jiangsu Yang-Yang Chemical Equipment Plant Inc</td>
			<td>CS-03-002</td>
			<td>1000L / 2000L</td>
		</tr>
		<tr>
			<td>heptane</td>
			<td>Nanjing Cook Biotechnology Co., Ltd.</td>
			<td>General Reagent</td>
			<td></td>
		</tr>
		<tr>
			<td>HPLC</td>
			<td>Guangzhou aoyi Technology Trading Co., Ltd</td>
			<td>LC-2030C 3D</td>
			<td></td>
		</tr>
		<tr>
			<td>Large scale rotary evaporators</td>
			<td>Guangzhou Xingshuo Instrument Co.,Ltd.</td>
			<td>RE-2002</td>
			<td></td>
		</tr>
		<tr>
			<td>Low temperature and constant temperature stirring reaction bath</td>
			<td>Guangzhou Yuhua Instrument Co., Ltd</td>
			<td>XHDHJF-3005</td>
			<td></td>
		</tr>
		<tr>
			<td>Low temperature coolant circulating pump</td>
			<td>Guangzhou Jincheng Scientific Instrument Co., Ltd</td>
			<td>XHDLSB-5/25</td>
			<td></td>
		</tr>
		<tr>
			<td>Megafuge 8R</td>
			<td>Thermo Scientific</td>
			<td>TS-HM8R</td>
			<td></td>
		</tr>
		<tr>
			<td>N, N-Diisopropyl ethylamine (DIPEA)</td>
			<td>Apicci Pharm</td>
			<td>General Reagent</td>
			<td></td>
		</tr>
		<tr>
			<td>N-dimethylformamide (DMF)</td>
			<td>Guangzhou bell Biotechnology Co., Ltd</td>
			<td>General Reagent</td>
			<td></td>
		</tr>
		<tr>
			<td>Octanoid acid</td>
			<td>Sigma-Aldrich</td>
			<td>O3907</td>
			<td></td>
		</tr>
		<tr>
			<td>Pd(OAc)2</td>
			<td>Xi&#39;an Catalyst New Materials Co.,ltd.</td>
			<td>200704</td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphate buffered saline</td>
			<td>Sigma-Aldrich</td>
			<td>1003335620</td>
			<td></td>
		</tr>
		<tr>
			<td>Potassium carbonate (K2CO3)</td>
			<td>Guangzhou Zhonghua Trade Co.,Ltd.</td>
			<td>General Reagent</td>
			<td></td>
		</tr>
		<tr>
			<td>Tert amyl alcohol</td>
			<td>Nanjing Cook Biotechnology Co., Ltd.</td>
			<td>General Reagent</td>
			<td></td>
		</tr>
		<tr>
			<td>tert-Butyl carbamate</td>
			<td>Nanjing Cook Biotechnology Co., Ltd.</td>
			<td>General Reagent</td>
			<td></td>
		</tr>
		<tr>
			<td>Thermo Mixer Heat/Cool</td>
			<td>KASVI</td>
			<td>K80-120R</td>
			<td></td>
		</tr>
		<tr>
			<td>toluene</td>
			<td>Liaoning cook Biotechnology Co., Ltd</td>
			<td>General Reagent</td>
			<td></td>
		</tr>
		<tr>
			<td>Vacuum drying oven</td>
			<td>Guangzhou Yuhua Instrument Co., Ltd</td>
			<td>DZF-6090</td>
			<td></td>
		</tr>
		<tr>
			<td>Water</td>
			<td>/</td>
			<td>/</td>
			<td></td>
		</tr>
		<tr>
			<td>Xantphos</td>
			<td>Liaoning cook Biotechnology Co., Ltd</td>
			<td>Asp20-44892</td>
			<td></td>
		</tr>
	</tbody>
</table>

scaled-up preparation of an intermediate of upatinib, act051-3

Upatinib, a Janus kinase inhibitor drug, was developed by a biotech company to treat immune diseases. The compound tert-butyl (5-toluenesulfonyl-5h-pyrrole [2,3-b] pyrazine-2-yl) carbamate (ACT051-3) is an important intermediate of Upatinib. To date, the steady industrial production of this intermediate compound (ACT051-3) has not been reported. In this study, we described the specific synthesis method and process of the compound ACT051-3 in terms of laboratory synthesis, pilot scale-up, and industrial production. During the exploration of the process route for ACT051-3, many appropriate adjustments and improvements were made to the reaction conditions, finally leading to the successful development of the optimal industrial production process for ACT051-3. The reaction time was nearly doubled by changing the state of the potassium carbonate involved in the reaction, which greatly improved the reaction efficiency. Additionally, by introducing N,N-diisopropylethylamine (DIPEA) to the reaction, the amount of the expensive catalyst Pd(OAc)2 was reduced 2.5-fold, significantly lowering production costs, confirming the feasibility of this process route and the industrial production of ACT051-3, and satisfying market demand for this important intermediate.

This study provides the scaled-up synthesis process for the important intermediate ACT051-3 of Upatinib (Figure 1 and Figure 2). The protocol section (steps 1-3) specifically shows the gram-grade synthesis, pilot-scale kilogram-grade synthesis, and scale-up production step of the compound ACT051-2 and the intermediate ACT051-3.
In the course of exploring the optimal route for the compound ACT051-2, as shown in <strong...

Watch this Scientific Journal Video about Scaled-Up Preparation of an Intermediate of Upatinib, ACT051-3 at JoVE.com

Scaled-Up Preparation of an Intermediate of Upatinib, ACT051-3

Here, we present a protocol for the scaled-up synthesis of the intermediate tert-butyl (5-toluenesulfonyl-5h-pyrrole [2,3-b] pyrazine-2-yl) carbamate (ACT051-3) of Upatinib.

Upatinib, a Janus kinase inhibitor drug, was developed by a biotech company to treat immune diseases. The compound tert-butyl ...

Here, a protocol for the scaled-up synthesis of an intermediate of upatinib, ACT051-3, is presented. This method is simple to operate, low cost, and environmentally friendly. To begin, prepare ACT051-2 by dissolving 50 grams of the compound 2-bromo-5-tosyl-5H-pyrrolo[2, 3-b]pyrazine in 15 milliliters of N, N-dimethylformamide, or DMF in a round-bottom flask.Add 65.3 grams of diisopropylethylamine to the reaction solution under nitrogen protection, and cool the temperature to 0 to 5 degrees Celsius through a cool water bath. Add 60.2 grams of para-toluene sulphonyl chloride, or TsCL, dissolved in 12 milliliters of DMF, and raise the temperature to 20 to 30 degrees Celsius through the warm water bath. Stir for about 1 hour.Add 600 milliliters of cool water to the mixture, and stir for another 1 hour. Filter the product under a vacuum using a glass funnel with a sanding board padded with filter paper. Wash with 200 milliliters of water several times, and dry, using an electric thermostatic drying oven to obtain pale yellow solid ACT051-2 with a yield of 78%For the synthesis of ACT051-3, dissolve 176.11 grams of ACT051-2 in 366.31 grams of 1, 4-dioxane in a three-port round-bottom flask, and add 65.75 grams of tert-butyl carbamate, 138.21 grams of granular potassium carbonate, 11.57 grams of Xantphos, and 2.25 grams of palladium acetate to the solution.Heat the mixture to 105 degrees Celsius, and stir for 7 hours under a nitrogen atmosphere. After cooling the mixture to room temperature, filter the product with a Buchner funnel. Wash the filter residue with 200 milliliters of ethyl acetate.Use a circulating water vacuum pump to concentrate the product under reduced pressure at 50 to 60 degrees Celsius, with a pressure value of minus 0.095 megapascal. Keep the pump working to maintain the pressure and obtain a dark brown oil. Purify the crude product with column chromatography, eluting with petroleum ether and ethyl acetate to obtain the target compound as a white solid with a yield of 93.5%For the pilot scale-up synthesis of ACT051-2, add one kilogram of 5.05 molar ACT051-1 and 1.305 kilograms of 10.1 molar DIPEA to a four-port round-bottom flask.Add 3 liters of DMF to the flask, and dissolve the solid. Heat the reaction mixture to 35 degrees Celsius. Add 1.203 kilograms of 6.31 molar TsCL to the reaction solution and stir for 1 hour.Stir the mixture until the completion of the reaction is confirmed by thin-layer chromatography, or TLC, and HPLC. Pour in 8.4 liters of cool water, and stir for another half an hour. Filter all liquids with a Buchner funnel, and rinse the crude product with 600 milliliters of water.Dry the resulting product at 70 degrees Celsius overnight using an electric thermostatic drawing oven, and obtain a product with a yield of 94.9%For the pilot scale-up synthesis of ACT051-3, add 3.31 liters of tert-amyl alcohol and 4.97 liters of toluene to the reaction kettle, and then add 1.66 kilograms of ACT051-2, 0.83 kilograms of tert-butyl carbamate, 1.301 kilograms of powdered potassium carbonate, 0.11 kilograms of Xantphos, and 0.31 kilograms of DIPEA to the solution. After evacuating nitrogen three times, as shown earlier, add 10 grams of palladium acetate to the reaction solution under nitrogen protection. Heat the reaction mixture to 90 degrees Celsius and stir for 4 hours.After that, cool the mixture to 40 degrees Celsius or below. Filter the reaction solution with a Buchner funnel using diatomite as a filter aid, and wash the filter cake with toluene. Collect and concentrate the filtrate using a circulating water vacuum pump, as shown earlier.Add 300 milliliters of heptane and stir for 20 minutes. After filtering the reaction solution again with a Buchner funnel, rinse the crude product with 50 milliliters of heptane. Dry and obtain the product with a yield of 96.3%The reaction route and conditions of the intermediate ACT051-3 before optimization and after optimization are presented in this figure.The effect of different forms of TsCL on the synthetic compound ACT051-2 is shown here. The results show that solid TsCL is more conducive to industrial production. The effect of adding TsCL at different temperatures on the synthesis of ACT051-2 is shown here.The product yield was increased from 97.49%to 98.44%when TsCL was added from 0 to 5 degrees Celsius to 23 to 35 degrees Celsius. The effect of different post-treatment water consumption on the synthesis of ACT051-2 is presented here. The optimal conditions were achieved at a post-treatment water volume of 6 milliliters per gram ACT051-2.This data shows the effect of adding DIPEA to the reaction for the synthesis of ACT051-3. The introduction of DIPEA reduced the amount of palladium acetate by a factor of 2.5. The effect of different states of potassium carbonate and different reaction solvents on the reaction of the synthetic compound ACT051-3 are presented here.The results show that by changing the state of potassium carbonate involved in the reaction, the reaction time was reduced from 7 hours to 3.5 hours. Additionally, by switching from tert-amyl alcohol/1, 4-dioxane to tert-amyl alcohol/toluene, the reaction time was shortened to 3 hours, and the product peak area increased from 84.22%to 88.52%The addition of DIPEA is critical for the pilot scale-up synthesis and industrial production of compound ACT051-3.

scaled-up-preparation-of-an-intermediate-of-upatinib-act051-3

Research

JoVE Journal

Chemistry

Synthesis of Antiviral Tetrahydrocarbazole Derivatives by Photochemical and Acid-catalyzed C-H Functionalization via Intermediate Peroxides (CHIPS)

The direct functionalization of C-H bonds is an important and long standing goal in organic chemistry. Such transformations can be very powerful in order to streamline synthesis by saving steps, time and material compared to conventional methods that require the introduction and removal of activating or directing groups. Therefore, the functionalization of C-H bonds is also attractive for green chemistry. Under oxidative conditions, two C-H bonds or one C-H and one heteroatom-H bond can be transformed to C-C and C-heteroatom bonds, respectively. Often these oxidative coupling reactions require synthetic oxidants, expensive catalysts or high temperatures. Here, we describe a two-step procedure to functionalize indole derivatives, more specifically tetrahydrocarbazoles, by C-H amination using only elemental oxygen as oxidant. The reaction uses the principle of C-H functionalization via Intermediate PeroxideS (CHIPS). In the first step, a hydroperoxide is generated oxidatively using visible light, a photosensitizer and elemental oxygen. In the second step, the N-nucleophile, an aniline, is introduced by Br&#248;nsted-acid catalyzed activation of the hydroperoxide leaving group. The products of the first and second step often precipitate and can be conveniently filtered off. The synthesis of a biologically active compound is shown.

Synthesis of Antiviral Tetrahydrocarbazole Derivatives by Photochemical and Acid-catalyzed C-H Functionalization via Intermediate Peroxides CHIPS

A two-step procedure for the synthesis of pharmaceutically active indole-derivatives by C-H functionalization with anilines is described, using photo- and Br&#248;nsted acid catalysis.

The direct functionalization of C-H bonds is an important and long standing goal in organic chemistry. Such transformations can be very powerful in order ...

Real-time Monitoring of Reactions Performed Using Continuous-flow Processing: The Preparation of 3-Acetylcoumarin as an Example

By using inline monitoring, it is possible to optimize reactions performed using continuous-flow processing in a simple and rapid way. It is also possible to ensure consistent product quality over time using this technique. We here show how to interface a commercially available flow unit with a Raman spectrometer. The Raman flow cell is placed after the back-pressure regulator, meaning that it can be operated at atmospheric pressure. In addition, the fact that the product stream passes through a length of tubing before entering the flow cell means that the material is at RT. It is important that the spectra are acquired under isothermal conditions since Raman signal intensity is temperature dependent. Having assembled the apparatus, we then show how to monitor a chemical reaction, the piperidine-catalyzed synthesis of 3-acetylcoumarin from salicylaldehyde and ethyl acetoacetate being used as an example. The reaction can be performed over a range of flow rates and temperatures, the in-situ monitoring tool being used to optimize conditions simply and easily.

Real-time monitoring allows for fast optimization of reactions performed using continuous-flow processing. Here the preparation of 3-acetylcoumarin is used as an example. The apparatus for performing in-situ Raman monitoring is described, as are the steps required to optimize the reaction.

By using inline monitoring, it is possible to optimize reactions performed using continuous-flow processing in a simple and rapid way. It is also possible ...

Preparation of Carbon Nanosheets at Room Temperature

Amphiphilic molecules equipped with a reactive, carbon-rich "oligoyne" segment consisting of conjugated carbon-carbon triple bonds self-assemble into defined aggregates in aqueous media and at the air-water interface. In the aggregated state, the oligoynes can then be carbonized under mild conditions while preserving the morphology and the embedded chemical functionalization. This novel approach provides direct access to functionalized carbon nanomaterials. In this article, we present a synthetic approach that allows us to prepare hexayne carboxylate amphiphiles as carbon-rich siblings of typical fatty acid esters through a series of repeated bromination and Negishi-type cross-coupling reactions. The obtained compounds are designed to self-assemble into monolayers at the air-water interface, and we show how this can be achieved in a Langmuir trough. Thus, compression of the molecules at the air-water interface triggers the film formation and leads to a densely packed layer of the molecules. The complete carbonization of the films at the air-water interface is then accomplished by cross-linking of the hexayne layer at room temperature, using UV irradiation as a mild external stimulus. The changes in the layer during this process can be monitored with the help of infrared reflection-absorption spectroscopy and Brewster angle microscopy. Moreover, a transfer of the carbonized films onto solid substrates by the Langmuir-Blodgett technique has enabled us to prove that they were carbon nanosheets with lateral dimensions on the order of centimeters.

We present the synthesis of an amphiphilic hexayne and its use in the preparation of carbon nanosheets at the air-water interface from a self-assembled monolayer of these reactive, carbon-rich molecular precursors.

Amphiphilic molecules equipped with a reactive, carbon-rich "oligoyne" segment consisting of conjugated carbon-carbon triple bonds self-assemble into ...

Facile Preparation of 4-Substituted Quinazoline Derivatives

Reported in this paper is a very simple method for direct preparation of 4-substituted quinazoline derivatives from a reaction between substituted 2-aminobenzophenones and thiourea in the presence of dimethyl sulfoxide (DMSO). This is a unique complementary reaction system in which thiourea undergoes thermal decomposition to form carbodiimide and hydrogen sulfide, where the former reacts with 2-aminobenzophenone to form 4-phenylquinazolin-2(1H)-imine intermediate, whilst hydrogen sulfide reacts with DMSO to give methanethiol or other sulfur-containing molecule which then functions as a complementary reducing agent to reduce 4-phenylquinazolin-2(1H)-imine intermediate into 4-phenyl-1,2-dihydroquinazolin-2-amine. Subsequently, the elimination of ammonia from 4-phenyl-1,2-dihydroquinazolin-2-amine affords substituted quinazoline derivative. This reaction usually gives quinazoline derivative as a single product arising from 2-aminobenzophenone as monitored by GC/MS analysis, along with small amount of sulfur-containing molecules such as dimethyl disulfide, dimethyl trisulfide, etc. The reaction usually completes in 4-6 hr at 160 &#186;C in small scale but may last over 24 hr when carried out in large scale. The reaction product can be easily purified by means of washing off DMSO with water followed by column chromatography or thin layer chromatography.

A protocol for facile preparation of 4-substituted quinazoline derivatives from 2-aminobenzophenones, thiourea and dimethyl sulfoxide is presented.

Reported in this paper is a very simple method for direct preparation of 4-substituted quinazoline derivatives from a reaction between substituted ...

Preparation of Biopolymer Aerogels Using Green Solvents

Although the first reports on aerogels made by Kistler1 in the 1930s dealt with aerogels from both inorganic oxides (silica and others) and biopolymers (gelatin, agar, cellulose), only recently have biomasses been recognized as an abundant source of chemically diverse macromolecules for functional aerogel materials. Biopolymer aerogels (pectin, alginate, chitosan, cellulose, etc.) exhibit both specific inheritable functions of starting biopolymers and distinctive features of aerogels (80-99% porosity and specific surface up to 800 m2/g). This synergy of properties makes biopolymer aerogels promising candidates for a wide gamut of applications such as thermal insulation, tissue engineering and regenerative medicine, drug delivery systems, functional foods, catalysts, adsorbents and sensors. This work demonstrates the use of pressurized carbon dioxide (5 MPa) for the ionic cross linking of amidated pectin into hydrogels. Initially a biopolymer/salt dispersion is prepared in water. Under pressurized CO2 conditions, the pH of the biopolymer solution is lowered to 3 which releases the crosslinking cations from the salt to bind with the biopolymer yielding hydrogels. Solvent exchange to ethanol and further supercritical CO2 drying (10 - 12 MPa) yield aerogels. Obtained aerogels are ultra-porous with low density (as low as 0.02 g/cm3), high specific surface area (350 - 500 m2/g) and pore volume (3 - 7 cm3/g for pore sizes less than 150 nm).

A new way for the production of biopolymer-based aerogels by carbon dioxide (CO2) induced gelation is shown. The technique utilizes pressurized carbon dioxide (5 MPa) for the production of biopolymer hydrogels and supercritical CO2 (12 MPa) to convert gels into aerogels. The only solvents needed besides CO2 are water and ethanol.

Although the first reports on aerogels made by Kistler1 in the 1930s dealt with aerogels from both inorganic oxides (silica and others) and biopolymers ...

Anionic Polymerization of an Amphiphilic Copolymer for Preparation of Block Copolymer Micelles Stabilized by &#960;-&#960; Stacking Interactions

In this study, an amphiphilic copolymer that includes a core-forming block with phenyl groups was synthesized by living anionic polymerization of phenyl glycidyl ether (PheGE) on methoxy-polyethylene glycol (mPEG-b-PPheGE). Characterization of the copolymer revealed a narrow molecular distribution (PDI &#60; 1.03) and confirmed the degree of polymerization of mPEG122-b-(PheGE)15. The critical micelle concentration of the copolymer was evaluated using an established fluorescence method with the aggregation behavior evaluated by dynamic light scattering and transmission electronic microscopy. The potential of the copolymer for use in drug delivery applications was evaluated in a preliminary manner including in vitro biocompatibility, loading and release of the hydrophobic anti-cancer drug doxorubicin (DOX). A stable micelle formulation of DOX was prepared with drug loading levels up to 14% (wt%), drug loading efficiencies &#62; 60% (w/w) and sustained release of drug over 4 days under physiologically relevant conditions (acidic and neutral pH, presence of albumin). The high drug loading level and sustained release is attributed to stabilizing &#960;-&#960; interactions between DOX and the core-forming block of the micelles.

Anionic Polymerization of an Amphiphilic Copolymer for Preparation of Block Copolymer Micelles Stabilized by &amp;#960;-&amp;#960; Stacking Interactions

The key steps of living anionic polymerization of phenyl glycidyl ether (PheGE) on methoxy-polyethylene glycol (mPEG-b-PPheGE) are described. The resulting block copolymer micelles (BCMs) were loaded with doxorubicin 14% (wt%) and sustained release of drug over 4 days under physiologically relevant conditions was obtained.

In this study, an amphiphilic copolymer that includes a core-forming block with phenyl groups was synthesized by living anionic polymerization of phenyl ...

Accessing Valuable Ligand Supports for Transition Metals: A Modified, Intermediate Scale Preparation of 1,2,3,4,5-Pentamethylcyclopentadiene

A reliable, intermediate scale preparation of 1,2,3,4,5-pentamethylcyclopentadiene (Cp*H) is presented, based on modifications of existing protocols that derive from initial 2-bromo-2-butene lithiation followed by acid mediated dienol cyclization. The revised synthesis and purification of the ligand avoids the use of mechanical stirring while still permitting access to significant quantities (39 g) of Cp*H in good yield (58%). The procedure offers other additional benefits, including a more controlled quench of excess lithium during the production of the intermediate heptadienols and a simplified isolation of Cp*H of sufficient purity for metallation with transition metals. The ligand was subsequently used to synthesize [Cp*MCl2]2 complexes of both iridium and ruthenium to demonstrate the utility of the Cp*H prepared and purified by our method. The procedure outlined herein affords substantial quantities of a ubiquitous ancillary ligand support used in organometallic chemistry while minimizing the need for specialized laboratory equipment, thus providing a simpler and more accessible entry point into the chemistry of 1,2,3,4,5-pentamethylcyclopentadiene.

Reliable, intermediate scale preparation of 1,2,3,4,5-pentamethylcyclopentadiene (Cp*H) is presented. The revised protocol for the synthesis and purification of the ligand minimizes the need for specialized laboratory equipment while simplifying reaction workups and product purification. Use of Cp*H in the synthesis of [Cp*MCl2]2 complexes (M = Ru, Ir) is also described.

A reliable, intermediate scale preparation of 1,2,3,4,5-pentamethylcyclopentadiene (Cp*H) is presented, based on modifications of existing protocols that ...

Preparation and Characterization of C60/Graphene Hybrid Nanostructures

Physical thermal deposition in a high vacuum environment is a clean and controllable method for fabricating novel molecular nanostructures on graphene. We present methods for depositing and passively manipulating C60 molecules on rippled graphene that advance the pursuit of realizing applications involving 1D C60/graphene hybrid structures. The techniques applied in this exposition are geared towards high vacuum systems with preparation areas capable of supporting molecular deposition as well as thermal annealing of the samples. We focus on C60 deposition at low pressure using a homemade Knudsen cell connected to a scanning tunneling microscopy (STM) system. The number of molecules deposited is regulated by controlling the temperature of the Knudsen cell and the deposition time. One-dimensional (1D) C60 chain structures with widths of two to three molecules can be prepared via tuning of the experimental conditions. The surface mobility of the C60 molecules increases with annealing temperature allowing them to move within the periodic potential of the rippled graphene. Using this mechanism, it is possible to control the transition of 1D C60 chain structures to a hexagonal close packed quasi-1D stripe structure.

Preparation and Characterization of C60/Graphene Hybrid Nanostructures

Here we present a protocol for the fabrication of C60/graphene hybrid nanostructures by physical thermal evaporation. Particularly, the proper manipulation of deposition and annealing conditions allow the control over the creation of 1D and quasi 1D C60 structures on rippled graphene.

Physical thermal deposition in a high vacuum environment is a clean and controllable method for fabricating novel molecular nanostructures on graphene. We ...

An Efficient Sample Preparation Method to Enhance Carbohydrate Ion Signals in Matrix-assisted Laser Desorption/Ionization Mass Spectrometry

Sample preparation is a critical process in mass spectrometry (MS) analysis of carbohydrates. Although matrix-assisted laser desorption/ionization (MALDI) MS is the method of choice in carbohydrate analysis, poor ion signal and data reproducibility of carbohydrate samples continue to be severe problems. For quantitative analysis of carbohydrates, an effective analytical protocol providing superior data quality is necessary. This video demonstrates sample preparation protocols to improve signal intensity and minimize data variation of carbohydrates in MALDI-MS. After drying and crystallization of sample droplets, the crystal morphology is reformed by methanol before mass spectrometric analysis. The enhancement in carbohydrate signal is examined with MALDI imaging mass spectrometry (IMS). Experimental results show that crystal reformation adjusts crystalline structures and redistributes carbohydrate analytes. In comparison with the dried droplet preparation method in conventional MALDI-MS, reforming carbohydrate crystal morphologies with methanol shows significantly better signal intensity, ion image distribution, and data stability. Since the protocols demonstrated herein do not involve changes in sample composition, they are generally applicable to various carbohydrates and matrixes.

A protocol for enhancing carbohydrate ion signals in MALDI mass spectrometry by reforming crystalline structures during sample preparation processes is demonstrated.

Sample preparation is a critical process in mass spectrometry (MS) analysis of carbohydrates. Although matrix-assisted laser desorption/ionization (MALDI) ...

Microfluidic Preparation of Liquid Crystalline Elastomer Actuators

This paper focuses on the microfluidic process (and its parameters) to prepare actuating particles from liquid crystalline elastomers. The preparation usually consists in the formation of droplets containing low molar mass liquid crystals at elevated temperatures. Subsequently, these particle precursors are oriented in the flow field of the capillary and solidified by a crosslinking polymerization, which produces the final actuating particles. The optimization of the process is necessary to obtain the actuating particles and the proper variation of the process parameters (temperature and flow rate) and allows variations of size and shape (from oblate to strongly prolate morphologies) as well as the magnitude of actuation. In addition, it is possible to vary the type of actuation from elongation to contraction depending on the director profile induced to the droplets during the flow in the capillary, which again depends on the microfluidic process and its parameters. Furthermore, particles of more complex shapes, like core-shell structures or Janus particles, can be prepared by adjusting the setup. By the variation of the chemical structure and the mode of crosslinking (solidification) of the liquid crystalline elastomer, it is also possible to prepare actuating particles triggered by heat or UV-vis irradiation.

This article describes the microfluidic process and parameters to prepare actuating particles from liquid crystalline elastomers. This process allows the preparation of actuating particles and the variation of their size and shape (from oblate to strongly prolate, core-shell, and Janus morphologies) as well as the magnitude of actuation.

This paper focuses on the microfluidic process (and its parameters) to prepare actuating particles from liquid crystalline elastomers. The preparation ...