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 cool water bath. For nitrogen protection, pump the pressure of the reactor to -0.75--0.8 MPa, then pass N2 to balance the pressure at 0.1 MPa.</li>
		<li>Add 60.20 g of TsCl dissolved in 12 mL of DMF (0.32 M). Raise the temperature to 20-30 &#176;C through the warm water bath and stir for about 1 h. Add cool water (600.0 mL, 0-10 &#176;C) to the mixture and stir for another 1 h.</li>
		<li>Filter the product under a vacuum using a glass funnel with a sanding board padded with filter paper. Wash with water (200.0 mL) several times and dry using an electric thermostatic drying oven to obtain a pale-yellow solid (ACT051-2) with a yield of 78%.</li>
	</ol>
	</li>
	<li>Synthesis of tert-butyl (5-toluenesulfonyl-5H-pyrrole [2,3-b] pyrazine-2-yl) carbamate (ACT051-3)
	<ol>
		<li>Dissolve 176.11 g of ACT051-2 in 366.31 g of 1,4-dioxane in a three-port round-bottom flask.</li>
		<li>Add 65.75 g of tert-butyl carbamate, 138.21 g of granular potassium carbonate (2.0 eq), 11.57 g of xantphos (0.04 eq), and 2.25 g of Pd(OAc)2 (1.28% wt) to the solution.</li>
		<li>Heat the mixture to 105 &#176;C and stir for 7 h under a nitrogen atmosphere. Allow the mixture to cool to room temperature and filter the product with a Buchner funnel (the filter paper aperture is 80-120 &#181;m).</li>
		<li>Wash the filter residue with ethyl acetate (200 mL). Use a circulating water vacuum pump to concentrate the product under reduced pressure at 50-60 &#176;C with a pressure value of -0.095 MPa. Keep the pump working to maintain the pressure and obtain a dark brown oil.</li>
		<li>Purify the crude product with column chromatography, eluting with petroleum ether and ethyl acetate (V/V, 10/1) to obtain the target compound as a white solid with a yield of 93.5%.</li>
	</ol>
	</li>
</ol>
2. Pilot scale-up synthesis of compounds ACT051-2 and ACT051-3
<ol>
	<li>Pilot scale-up synthesis of 2-bromo-5-tosyl-5H-pyrrolo[2,3-b]pyrazine (ACT051-2)
	<ol>
		<li>Add 1.0 kg of ACT051-1 (5.05 M) and 1.305 kg of DIPEA (10.1 M) to a three-port round-bottom flask. Add 3 L of DMF (3 V) to the flask and dissolve the solid. Heat the reaction mixture to 35 &#176;C.</li>
		<li>Add 1.203 kg of TsCl (6.31 M) to the reaction solution and stir for 1 h. Stir the mixture until the completion of the reaction is confirmed by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC). In brief, take a small amount of reaction product and subject this to TLC monitoring. When the TLC shows that there is almost no raw material left, send the sample to the HPLC central control, and detect the reaction to raw material or the product ratio to be 0.1/99.9.</li>
		<li>Pour in 8.4 L of cool water and stir for another 0.5 h. Filter all liquids with a Buchner funnel (the filter paper aperture is 80-120 &#181;m) and rinse the crude product with 600 mL of water.</li>
		<li>Dry the resulting product at 70 &#176;C overnight using an electric thermostatic drying oven and obtain a product with a yield of 94.9%.</li>
	</ol>
	</li>
	<li>Pilot scale-up synthesis of tert-butyl (5-toluenesulfonyl-5H-pyrrole [2,3-b] pyrazine-2-yl) carbamate (ACT051-3)
	<ol>
		<li>Add 3.31 L of tert-amyl alcohol and 4.97 L of toluene (V/V, 2/3) to the reaction kettle.</li>
		<li>To the solution, add 1.66 kg of ACT051-2, 0.83 kg of tert-butyl carbamate, 1.301 kg of powdered potassium carbonate, 0.11 kg of xantphos, and 0.31 kg of DIPEA.</li>
		<li>Evacuate nitrogen three times, as in step 1.1.2, repeat this 3x, and add 10 g of Pd(OAc)2 (0.60% wt) to the reaction solution under nitrogen protection.</li>
		<li>Heat the reaction mixture to 90 &#176;C and stir for 4 h. Cool the mixture to 40 &#176;C or below.</li>
		<li>Filter the reaction solution with a Buchner funnel (the filter paper aperture is 80-120 &#181;m) using diatomite as a filter aid and washing the filter cake with toluene.</li>
		<li>Collect and concentrate the filtrate. Use a circulating water vacuum pump to concentrate the filtrate under reduced pressure at 50-60 &#176;C with a pressure value of -0.095 MPa. Keep the pump working to maintain the pressure.</li>
		<li>Add 300 mL of heptane and stir for 20 min. Filter the reaction solution again with a Buchner funnel (the filter paper aperture is 80-120 &#181;m) and rinse the crude product with heptane (50 mL, three times). Dry and obtain the product with a yield of 96.3%.</li>
	</ol>
	</li>
</ol>
3. Industrial production of compounds ACT051-2 and ACT051-3
<ol>
	<li>Industrial production of 2-bromo-5-tosyl-5H-pyrrolo[2,3-b]pyrazine (ACT051-2)
	<ol>
		<li>Confirm that the reactor is clean and free of water, and make sure the mixing device makes no noise when turning on and the bottom discharge valve of the reactor has been closed.</li>
		<li>Add 355.50 kg of DMF (3 V) into the 2,000 L enamel reactor and start to stir. Add 125.04 kg of ACT051-1 (1.0 eq) and 164.8 kg of DIPEA (2.0 eq) to the solution.</li>
		<li>Drop the temperature to 20-25 &#176;C under nitrogen protection. Add 150.22 kg of TsCl (1.25 eq) at 25-35 &#176;C in 10 batches within 3 h.</li>
		<li>Allow the mixture to remain at 25-35 &#176;C and stir for 2 h. Prepare 750.20 kg of cool water (3-4 &#176;C) in another 2,000 L reactor.</li>
		<li>Monitor the reaction by HPLC and confirm completion of the reaction when the remaining raw material is 0.5%.</li>
		<li>Add the cool water ( 3-4 &#176;C) to the reaction mixture and stir at 15-30 &#176;C for 1.5 h. Collect the filtrate and rinse with water (250-500 kg, 2-4 V) until it becomes neutral (tested using pH paper).</li>
		<li>Dry the product at 60-65 &#176;C for 30.5 h to obtain a light brown solid with a yield of 95.5%.</li>
	</ol>
	</li>
	<li>Industrial production of tert-butyl (5-toluenesulfonyl-5H-pyrrole [2,3-b] pyrazine-2-yl) carbamate (ACT051-3)
	<ol>
		<li>Confirm the reactor is clean and free of water, and make sure the mixing device is normal and the bottom discharge valve of the reactor has been closed.</li>
		<li>Add 340.10 kg of tert-amyl alcohol (2 V) and 552.50 kg of toluene (3 V) into a 2,000 L enamel reactor and start to stir.</li>
		<li>Add 212.40 kg of ACT051-2 (1.0 eq), 106.00 kg of tert-butyl carbamate (1.5 eq), 166.70 kg of powdered potassium carbonate (2.0 eq), 14.10 kg of xantphos (0.04 eq), 39.40 kg of DIPEA (0.5 eq), and 1.06 kg of Pd(OAc)2 (0.5% wt) to the solution.</li>
		<li>Replace the nitrogen four times, allow the mixture to heat to 85-95 &#176;C under nitrogen protection, and stir for 3 h. Monitor the reaction by HPLC and confirm completion of the reaction when the remaining raw material is 0.44%.</li>
		<li>Cool the reaction temperature to 20-30 &#176;C. Collect the solution in batches and place it in a 125 L plastic bucket for 30 min.</li>
		<li>Filter the reaction solution with a chemical suction filter barrel (the filter bag aperture is 10-15 &#181;m) using diatomite as a filter aid and washing the filter cake with toluene (185.20-370.40 kg, 1-2 V).</li>
		<li>Collect the filtrate in an enamel vessel and use a circulating water vacuum pump under reduced pressure at 55-65 &#176;C for 10 h with a pressure value of -0.095 MPa. Keep the pump working to maintain the pressure and obtain the product compound as a viscous liquid.</li>
		<li>Evaporate the product twice by pumping toluene and continuing the reaction. Concentrate the final product for 4 h. Purify the resulting product with column chromatography, eluting with heptane and ethyl acetate (V/V, 10/1-3/1), to obtain a solid with a yield of 98.5%.</li>
	</ol>
	</li>
</ol>

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The authors have nothing to disclose.

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 increases the amount of solvent in the reaction system, resulting in more waste fluid. As a result, we chose solid TsCl for the reaction in the pilot scale-up and industrial production (steps 2.1.2 and 3.1.3 of the protocol, respectively) and obtained good experimental results, keeping in mind the concept of environmental protection and green chemistry (Table 1).
In addition, it was found in the pilot scale-up experiments that adding a large amount of TsCl at a low temperature makes the reaction system too viscous to be stirred, resulting in the risk of encapsulation of the raw material. To address this risk, the temperature was increased from 0-5 &#176;C to 25-35 &#176;C (steps 2.1.1 or 3.1.4 of the protocol), sufficient stirring was obtained for the solid TsCl added, the reaction solution system had good fluidity, and the reaction proceeded smoothly (Table 2).
Moreover, a large amount of water (15 mL/g ACT051-1) was required for the post-treatment experiments of ACT051-2, which was not suitable for the scaling up production experiments and generated more waste solution. Therefore, from the perspective of environmental protection, the optimal amount of water used for post-treatment was immediately investigated in the exploration of the production process conditions. As shown in Table 3, when the amount of post-treatment water was reduced to 6 mL/g ACT051-1, the yield of the experiment showed a little impact, but could greatly improve the efficiency and reduce the generation of waste liquid.
In exploring the industrial production process route of the intermediate ACT051-3, we conducted a large number of process optimization experiments and solved many scaling problems. In the synthesis of ACT051-3, ACT051-2 underwent a Buchwald-Hartwig coupling reaction with tert-butyl carbamate catalyzed by Pd(OAc)2 and xanthphos to give the compound ACT051-3. Pd(0) is an active species of palladium commonly used in coupling responses like Suzuki and Buchwald16,17,18,19. However, Pd(II) is commonly used to catalyze the reaction. As a result, we used an amine compound (DIPEA) to reduce Pd(II) to Pd(0), leaving the catalytic system of the entire reaction in a Pd(0) and Pd(II) catalytic cycle. With the addition of DIPEA, the amount of the expensive catalyst Pd(OAc)2 was greatly reduced from 1.28% to 0.5% (Table 4), which greatly reduced the cost of scale-up production and further strengthened the rationality and feasibility of the method.
The long reaction time brings about a large number of side reactions and generates impurities, which is not conducive to scaling up production. Through extensive laboratory studies, it was found that changing the state of potassium carbonate from granular to powder (steps 1.2.2 or 2.2.2 of the protocol) resulted in an appropriate reduction of the reaction time, which is more favorable for industrial production (Table 5). In addition, considering that the 1,4-dioxane used in the small-scale pilot experiment was more difficult to concentrate (step 1.2.1 of the protocol), we considered the combination of toluene and tert-butanol (steps 2.2.1 or 3.2.2 of the protocol; V/V, 2/3), which are better concentrated solvents, as reaction solvents for the industrial production. The results showed that the reaction obtained a great promotion rate, reduced the reaction time, and improved the efficiency of the reaction (see Table 6).
In conclusion, after repeated exploration of the experimental conditions, the optimal process scale-up conditions for compounds ACT051-2 and ACT051-3 were finally obtained, and the process flow diagram of this scale-up production is shown in Figure 2. The results of the scaled-up production showed that the whole process was stable, and the product yield was normal (steps 3.1.7 or 3.2.8 of the protocol).
The industrial production route obtained in this study is a new route available, and the feasibility of this process route in future industrial production was also confirmed. In addition, the results obtained in this study provide some technical research basis for future research of the industrial production route of compounds ACT051-2 and ACT051-3.
However, there is still room for optimization and improvement of the process route, such as the amount of TsCl (1.25 eq), which is excessive in the synthesis of ACT051-2 and can be further reduced. In addition, in the synthesis of ACT051-3, only toluene can be used as the reaction solvent to facilitate post-processing, and the amount of catalyst Pd(OAc)2 may continue to decrease. The above technical issues can be further studied and explored in future work to better scale-up the production of the synthetic compound ACT051-3.

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 Table 1, it was found that solid TsCl was more involved in the reaction than liquid TsCl (dissolved in DMF, step 3.1 of the protocol) and significantly reduced the amount of DMF by nearly threefold. In addition, the product yield was increased from 97.49% to 98.44% by increasing the temperature of the mixed solution when TsCl was added from 0-5 &#176;C to 23-35 &#176;C (shown in Table 2). Further, experiments on the consumption of post-treatment water were conducted. As shown in Table 3, after a 2.5-fold reduction in water consumption (from 15 mL/g ACT051-2 to 6 mL/g ACT051-2), the reaction yield decreased by 2.5%, but the waste solution generation was reduced, and the reaction efficiency was significantly improved.
A series of experimental conditions were developed to obtain the optimized process route for the intermediate ACT051-3. As shown in Table 4, by introducing DIPEA into the reaction and replacing the reaction solvent with tert-amyl alcohol/toluene (V/V, 2/3), the amount of Pd(OAc)2 was reduced 2.5-fold (from 1.28% wt to 0.5% wt), which significantly reduced the production cost and further improved the feasibility of scaling up the production. Additionally, by changing the state of K2CO3 involved in the reaction, the reaction time was reduced from 7 h to 3.5 h, which greatly improved the reaction efficiency (as shown in Table 5). Additionally, by switching from tert-amyl alcohol/1,4-dioxane (V/V, 1/4) to tert-amyl alcohol/toluene (V/V, 2/3), the reaction time was shortened to 3 h, the product peak area increased from 84.22% to 88.52%, and the time it took for the product to concentrate was significantly shortened, all of which improved the reaction&#39;s efficiency (see Table 6).
Both compounds ACT051-2 and ACT051-3 were chemically characterized by proton nuclear magnetic resonance (1H NMR), HPLC, and high-resolution mass spectrometry. The analysis methods (HPLC, 1H NMR, and electrospray ionization [ESI] spectroscopy) of ACT051-2 and ACT051-3 can be found in the supporting work (Supplementary Table 1, Supplementary Figure 1, Supplementary Figure 2, Supplementary Figure 3, Supplementary Figure 4, Supplementary Figure 5, and Supplementary Figure 6). The characterization data for ACT051-2 and ACT051-3 are reported below:
2-bromo-5-tosyl-5H-pyrrolo[2,3-b]pyrazine (ACT051-2): 
1H NMR (500 MHz, DMSO-d6)&#948;8.59 (s,1H), 8.37 (d, J = 4.1 Hz, 1H), 8.00 (d, J = 8.2 Hz, 2H), 7.46 (d, J = 8.2 Hz, 2H), 7.02 (d, J = 4.0 Hz, 1H), 3.29 (d, J = 11.9 Hz, 3H). ESI: m/z calculated for C13H10BrN3O2S [M] + 352.21, found to be 352.00.
Tert-butyl (5-toluenesulfonyl-5H-pyrrole [2,3-b] pyrazine-2-yl) carbamate (ACT051-3): 
1H NMR (500 MHz, CDCl3) &#948;8.98 (s, 1H), 7.95 (d, J = 8.4 Hz, 2H), 7.84 (d, J = 4.1 Hz, 1H), 7.21 (s, 1H), 7.19-7.17 (m, 1H), 6.53 (d, J = 4.1 Hz, 1H), 2.30 (s, 3H), 1.45 (s, 9H). ESI: m/z calculated for C18H20N4O4S [M+H] + 389.12, found to be389.15.
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/64514/64514fig01.jpg" /> 
Figure 1: Synthesis route of the intermediate ACT051-3. (A) The reaction route and conditions of ACT051-3 before optimization: i) DMF, DIPEA, TsCl; ii) xantphos, Pd(OAc)2, K2CO3, tert-amyl alcohol/1,4-dioxane (V/V, 1/4); (B) The reaction route and conditions of ACT051-3 after optimization: i) DMF, DIPEA, TsCl; ii) xantphos, Pd(OAc)2, K2CO3, DIPEA, tert-amyl alcohol/toluene (V/V, 2/3). <a href="https://www.jove.com/files/ftp_upload/64514/64514fig01large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/64514/64514fig02.jpg" /> 
Figure 2: The process flow diagram of compounds ACT051-2 and ACT051-3 in scale-up production. (A) Process flow diagram of ACT051-2 in scale-up production. (B) Process flow diagram of ACT051-3 in scale-up production. <a href="https://www.jove.com/files/ftp_upload/64514/64514fig02large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Number</td>
			<td>State of TsCl</td>
			<td>V(DMF) (Laboratory test)</td>
			<td>V(DMF) (Pilot scale up)</td>
		</tr>
		<tr>
			<td>1</td>
			<td>dissolve in DMF</td>
			<td>8.5 V</td>
			<td>54 V</td>
		</tr>
		<tr>
			<td>2</td>
			<td>solid</td>
			<td>3.0 V</td>
			<td>18 V</td>
		</tr>
	</tbody>
</table>
Table 1: Effect of different forms of TsCl on the synthetic compound ACT051-2. The different states of TsCl include liquid TsCl (dissolved in DMF) and solid TsCl. Experimental results show that solid TsCl is more conducive to industrial production.
<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Number</td>
			<td>Temperature (&#176;C)</td>
			<td>Condition of mix</td>
			<td>Whether has a clarification process</td>
			<td>Yield</td>
			<td>Purity</td>
		</tr>
		<tr>
			<td>1</td>
			<td>0-5</td>
			<td>viscous</td>
			<td>No</td>
			<td>97.49%</td>
			<td>96.85%</td>
		</tr>
		<tr>
			<td>2</td>
			<td>25-35</td>
			<td>good mixing</td>
			<td>Yes</td>
			<td>98.44%</td>
			<td>96.99%</td>
		</tr>
	</tbody>
</table>
Table 2: Effect of adding TsCl at different temperatures on the synthesis of ACT051-2. Adding TsCl to the reaction at 0-5 &#176;C or 23-35 &#176;C.
<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Number</td>
			<td>Water consumption</td>
			<td>Yield</td>
			<td>Purity</td>
		</tr>
		<tr>
			<td>1</td>
			<td>15 mL / g ACT051-1</td>
			<td>97.49%</td>
			<td>96.85%</td>
		</tr>
		<tr>
			<td>2</td>
			<td>6 mL / g ACT051-1</td>
			<td>94.90%</td>
			<td>97.69%</td>
		</tr>
		<tr>
			<td>3</td>
			<td>9 mL / g ACT051-1</td>
			<td>95.07%</td>
			<td>96.71%</td>
		</tr>
	</tbody>
</table>
Table 3: Effect of different post-treatment water consumption on the synthesis of ACT051-2. Try the different post-treatment water consumptions, including 15 mL/g ACT051-1, 9 mL/g ACT051-1, and 6 mL/g ACT051-1. The optimal conditions were achieved at a post-treatment water volume of 6 mL/g ACT051-2.
<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Number</td>
			<td>Equivalent of DIEPA</td>
			<td>Equivalent of K2CO3</td>
			<td>Equivalent of Pd(OAc)2</td>
		</tr>
		<tr>
			<td>1</td>
			<td>0.0 eq</td>
			<td>3.0 eq</td>
			<td>1.28% wt</td>
		</tr>
		<tr>
			<td>2</td>
			<td>2.0 eq</td>
			<td>2.0 eq</td>
			<td>0.60% wt</td>
		</tr>
		<tr>
			<td>3</td>
			<td>1.0 eq</td>
			<td>2.0 eq</td>
			<td>0.60% wt</td>
		</tr>
		<tr>
			<td>4</td>
			<td>0.5 eq</td>
			<td>2.0 eq</td>
			<td>0.60% wt</td>
		</tr>
	</tbody>
</table>
Table 4: Effect of adding DIPEA to the reaction for the synthesis of ACT051-3. Exploring the effect of DIPEA addition or not on the reaction. The results showed that the introduction of DIPEA reduced the amount of Pd(OAc)2 by a factor of 2.5 (from 1.28% wt to 0.5% wt).
<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Number</td>
			<td>State of K2CO3</td>
			<td>Equivalent</td>
			<td>Reaction time (h)</td>
		</tr>
		<tr>
			<td>1</td>
			<td>solid</td>
			<td>2.0 eq</td>
			<td>7</td>
		</tr>
		<tr>
			<td>2</td>
			<td>powdered</td>
			<td>2.0 eq</td>
			<td>3.5</td>
		</tr>
	</tbody>
</table>
Table 5: Effect of different states of K2CO3 on the reaction of the synthetic compound ACT051-3. Select potassium carbonate in granular or powder form to participate in the reaction.
<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Number</td>
			<td>Dosage of Pd(OAc)2</td>
			<td>Reaction solvent</td>
			<td>V/V</td>
			<td>Reaction time / h</td>
			<td>Product&#160; peak area/ %</td>
		</tr>
		<tr>
			<td>1</td>
			<td>0.60% wt</td>
			<td>tert amyl alcohol / 1,4-dioxane</td>
			<td>1-4</td>
			<td>3.5</td>
			<td>84.22</td>
		</tr>
		<tr>
			<td>2</td>
			<td>0.60% wt</td>
			<td>tert amyl alcohol / 1,4-dioxane</td>
			<td>2-3</td>
			<td>3.5</td>
			<td>83.34</td>
		</tr>
		<tr>
			<td>3</td>
			<td>0.60% wt</td>
			<td>tert amyl alcohol / Toluene</td>
			<td>2-3</td>
			<td>3</td>
			<td>88.52</td>
		</tr>
		<tr>
			<td>4</td>
			<td>0.50% wt</td>
			<td>tert amyl alcohol / Toluene</td>
			<td>2-3</td>
			<td>2.25</td>
			<td>87.11</td>
		</tr>
	</tbody>
</table>
Table 6: Effect of different reaction solvents on the reaction of the synthetic compound ACT051-3. Tert-amyl alcohol/1,4-dioxane (V/V, 1/4) and tert-amyl alcohol/toluene (V/V, 2/3) are selected as reaction resolvents.
Supplementary Table 1: Analytical method of compounds ACT051-2 and ACT051-3. Specific chromatographic conditions for the analysis of compounds ACT051-2 and ACT051-3, including the instrument, method name, liquid phase column, mobile phase, column temperature, current speed, and wavelength. <a href="https://www.jove.com/files/ftp_upload/64514/Supplementary table 1_RE.xlsx" target="_blank">Please click here to download this File.</a>
Supplementary Figure 1: The high-performance liquid chromatograms of ACT051-2. The results for the data were detected by HPLC. <a href="https://www.jove.com/files/ftp_upload/64514/Supplementary Figure 1_RE.png" target="_blank">Please click here to download this File.</a>
Supplementary Figure 2: The high-performance liquid chromatograms of ACT051-3. The results for the data were detected by HPLC. <a href="https://www.jove.com/files/ftp_upload/64514/Supplementary Figure 2_RE.png" target="_blank">Please click here to download this File.</a>
Supplementary Figure 3: MS spectrum of ACT051-2. The results for the data were detected by ESI spectroscopy. <a href="https://www.jove.com/files/ftp_upload/64514/Supplementary Figure 3_RE.png" target="_blank">Please click here to download this File.</a>
Supplementary Figure 4: MS spectrum of ACT051-3. The results for the data were detected by ESI spectroscopy. <a href="https://www.jove.com/files/ftp_upload/64514/Supplementary Figure 4_RE.png" target="_blank">Please click here to download this File.</a>
Supplementary Figure 5: 1H NMR spectrum of ACT051-2. The results for the data were analyzed using MestReNova. <a href="https://www.jove.com/files/ftp_upload/64514/Supplementary Figure 5_RE.png" target="_blank">Please click here to download this File.</a>
Supplementary Figure 6: 1H NMR spectrum of ACT051-3. The results for the data were analyzed using MestReNova. <a href="https://www.jove.com/files/ftp_upload/64514/Supplementary Figure 6_RE.png" target="_blank">Please click here to download this File.</a>

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 ...

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

Nanyang Technological University

Research

JoVE Journal

Chemistry

915 Views.  Aneo Chem-Tech Co., Ltd. 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.

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

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.

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 ...

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

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